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 AppliquÈes 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:
ï Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)
ï Advances in instrumentation Co-chairs: M. Ellisman (UCSD); Bram Koster (Univ. ofUltrect, Netherlands)
ï 3D reconstruction algorithms Co-chairs: N. Volkmann (Burnham Institute); TBA
ï Visualization & quantitative analysis Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)
ï Moving tomography to the mainstream: data sharing, data integration, & model building Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)
ï Emerging technologies for the multiscale: cell to tissue and molecule to cell Co-chairs: D. Hanein (Burnham Institute); C. Larabell (UCSF)
Sessions will include both invited speakers and talks selected from submitted abstracts.
For further information please check out our web site: http://www.4icet.org
or contact Mark H. Ellisman, Lead Congress Organizer (mark-at-ncmir.ucsd.edu) or Grace Osborne, Congress Coordinator (gosborne-at-ncmir,ucsd.edu)
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.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:
ï Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)
ï Advances in instrumentation Co-chairs: M. Ellisman (UCSD); Bram Koster (Univ. ofUltrect, Netherlands)
ï 3D reconstruction algorithms Co-chairs: N. Volkmann (Burnham Institute); TBA
ï Visualization & quantitative analysis Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)
ï Moving tomography to the mainstream: data sharing, data integration, & model building Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)
ï Emerging technologies for the multiscale: cell to tissue and molecule to cell Co-chairs: D. Hanein (Burnham Institute); C. Larabell (UCSF)
Sessions will include both invited speakers and talks selected from submitted abstracts.
For further information please check out our web site: http://www.4icet.org
or contact Mark H. Ellisman, Lead Congress Organizer (mark-at-ncmir.ucsd.edu) or Grace Osborne, Congress Coordinator (gosborne-at-ncmir,ucsd.edu)
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
SociÈtÈ 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
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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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 ñ 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.
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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)-NanogoldÆ]
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 NiñNTA 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 **********************************************
==============================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==============================
==============================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 £60 DX5 microscope has 250x magnification and a CMOS 'camera').
However, from my experience even going to £2,000 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 £12k to £45 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 £70k 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 £20k to £70k.
You can get pretty good results from the new breed of sub-£1,000 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 £300 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 (£260) 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 (£300). 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 ¼ 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
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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 £1,000'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 £60 DX5 microscope has 250x magnification and a CMOS 'camera').
However, from my experience even going to £2,000 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 £12k to £45 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 £70k 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 £20k to £70k.
You can get pretty good results from the new breed of sub-£1,000 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 £300 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 (£260) 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 (£300). 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 ¼ 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 £2k (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 £900, and LS 9000ED £2500) 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 £500, 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 £100 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 doesn¥t 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,
TÈc. Ppal Elbio MartÌnez 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 ìEhrlich 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 ìEM of Fully Hydrated Samplesî
Eva Chi, PhD, University of Chicago ìRole of Cell Membrane in the Pathogenisis of Alzheimerís Diseaseî
Alex Kandel, PhD, University of Notre Dame ìStructure 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
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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 } - ∑. } } 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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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 £400 to £550 [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 £280 in 2006) produces digitised images scanned at maximum resolution that are distinguishable to those from our Agfa DuoScan flatbed scanner (that cost £5,000 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 £5 to £10 (} 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 (£260) 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 £200 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 £280 in 2006) produces digitised images scanned at maximum resolution that are indistinguishable to those from our Agfa DuoScan flatbed scanner (that cost £5,000 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 ìon-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 ìon-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
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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.
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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 £60) 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-Ångstrom 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.
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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.
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 ³out of the box² 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 (4M¹s)
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 ³care and feeding² 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 ³out of the box² 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 (4M¹s) } } 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 ³care and feeding² 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, UniversitÈ 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 (20Å) onto MgO specimens prior to TEM examination. If the film grown on the MgO is not very conductive, then I coat both sides. In your case you have a conductive layer on one side (Fe), so you may get away with coating just the MgO side.
I hope this helps.
Regards
Dave Mitchell -- Dr David Mitchell ANSTO Materials PMB 1 Menai NSW 2234 Australia
tel 61 2 9717 3456 fax 61 2 9543 7179
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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 56ƒC) at room temperature (about 18ƒC) with a microtome flat razor blade. Thank you in advance. Best Regards,
Good morning, everybody I am looking for advice on defining resolution and minimal feature size through information theory. In 1992, there was a paper by D. Van Dyck and A.F. de Jong (Ultramicroscopy 47, 266 (1992)), which addressed in some detail general principles on information theory as applied to image formation mechanism in electron microscopy. I am wondering what the current status of this field is. Thank you in advance Sergei
-- Sergei V. Kalinin, Oak Ridge National Laboratory, 1 Bethel Valley Rd, Bldg. 3025, MS6030, Oak Ridge, TN 37831 Phone: (865) 241-0236 FAX: (865) 574-4143 URL: sergei2.kalininweb.com New: http://nanotransport.ornl.gov
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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:
ï Imaging of dynamic structures and correlative microscopy Co-Chairs: O. Medalia (Ben-Gurion Univ.); Jack Johnson (The Scripps Research Institute)
ï Advances in instrumentation Co-chairs: M. Ellisman (UCSD); Bram Koster (Leiden University Medical Center, Netherlands)
ï 3D reconstruction algorithms Co-chairs: N. Volkmann (Burnham Institute); Michael Rademacher (Univ. of Vermont)
ï Visualization & quantitative analysis Co-chairs: R. Whitaker (Univ. of Utah); D. Mastronarde (Univ. of Colorado)
ï Moving tomography to the mainstream: data sharing, data integration, & model building Co-chairs: M. Martone (UCSD); J-M. Carazo (Univ. of Madrid)
ï Emerging technologies for the multiscale: cell to tissue and molecule to cell Co-chairs: D. Hanein (Burnham Institute); C. Larabell (UCSF)
Sessions will include both invited speakers and talks selected from submitted abstracts.
For further information please check out our web site: http://www.4icet.org or contact Mark H. Ellisman, Lead Congress Organizer (mark-at-ncmir.ucsd.edu) or Grace Osborne, Congress Coordinator (gosborne-at-ncmir,ucsd.edu)
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both temanalysis-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] TEM/FIB: FEI 200 single beam FIB question
Question: Hi: We are thinking of working a group that wants to add FIB capabilities to their lab to prepare semiconductor samples. They are looking into purchasing a pre-owned FEI 200 (single beam) with a Magnum column.
I have talked to a few of my contacts and they believe this instrument would be best suited to prepare TEM samples of larger technologies at best. They believe something like a pre-owned dual beam FEI 820 or similar might be a better choice for general TEM sample preparation. I don't really know the answer, but wanted to get actual users opninions.
Ideally, I would like to receive feedback of actual users of the FEI 200 with the Magnum column and see if they can easily prepare semiconductor TEM samples with smaller technologies and: 1) Approximately how long it takes to make a sample 2) What size geometries they can easily and routinely section.
If you do not use this particular instrument, but are a frequent FIB user and have a general opinion, I would like to hear from you also.
Thanks for being a great information resource, Sandra Keller
I am currently using our Zeiss axiovert 200M to observe cell comparments (mitochondria, golgi, endosomes,...) by fluorescence. For this purpose, the main "usable" objectives available are: - 40x/0,50 LD A-Plan - 100x/1,3 Oil EC Plan Neofluar
The 40x LD is useful for our life cell imaging experiments, so I don't think we should change it. But I wonder if the 100x is really the best solution. I must say that I am a little bit disappointed by the quality of my images (I also try to improve the preparation of the samples!). When i look to the "images samples" on the CD which was included with the axiovision soft, I notice that most of the pictures of cells are taken with a 63x apochromat. I searched the site of Zeiss and found an interesting 63x apochromat with a NA of 1.4 and corrected for coverglass thickness (0,17). What is your experience with the different Zeiss objectives?
My second question concerns the Apotome feature of Axiovision. It seems to do the same job as 3D deconvolution, but how does it work? Why choose one or the other?
Thank you all in advance.
Stephane-without-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Concerning the objectives, you might want to consider buying an EC Plan-NeoFluar 40x 1,3 oil. A great objective for fluorescence (offers a clear & intense image) that I even very often prefer instead of our 100x oil.
The ApoTome is based on the grid projection theory (more details can be found on the Zeiss webpage). The difference with Deconvolution software is that this device (hardware) shows you immediately, while acquiring the image, the result. The deconvolution software will only show you the result after acquisition and running the program. If than your image does not turn out nice, you'll have to restart from the acquisition on, whereas with the ApoTome, you can immediately see the result and restart if necessary.
Hope it helps a bit!
Best,
Sven Terclavers
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: vrijdag 7 april 2006 10:58 To: sven.terclavers-at-med.kuleuven.be
Dear listers,
I am currently using our Zeiss axiovert 200M to observe cell comparments (mitochondria, golgi, endosomes,...) by fluorescence. For this purpose, the main "usable" objectives available are: - 40x/0,50 LD A-Plan - 100x/1,3 Oil EC Plan Neofluar
The 40x LD is useful for our life cell imaging experiments, so I don't think we should change it. But I wonder if the 100x is really the best solution. I must say that I am a little bit disappointed by the quality of my images (I also try to improve the preparation of the samples!). When i look to the "images samples" on the CD which was included with the axiovision soft, I notice that most of the pictures of cells are taken with a 63x apochromat. I searched the site of Zeiss and found an interesting 63x apochromat with a NA of 1.4 and corrected for coverglass thickness (0,17). What is your experience with the different Zeiss objectives?
My second question concerns the Apotome feature of Axiovision. It seems to do the same job as 3D deconvolution, but how does it work? Why choose one or the other?
Thank you all in advance.
Stephane-without-i
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
Hello, Interested if anyone knows of a substitute for poly-L-lysine in TEM. I am currently using it to coat carbon coated formvar coated Ni grids and examine plasma membranes with immunolabelling. Fingers crossed. Martyn.
Martyn Quinn Henry Wellcome Laboratory for Cell Biology Division of Biochemistry & Molecular Biology Davidson Building Faculty of Biomedical and Life Sciences University of Glasgow G12 8QQ
==============================Original Headers============================== 1, 21 -- From mq7x-at-udcf.gla.ac.uk Fri Apr 7 05:26:35 2006 1, 21 -- Received: from hillhead.cent.gla.ac.uk (hillhead.cent.gla.ac.uk [130.209.16.101]) 1, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k37AQZLs032136 1, 21 -- for {microscopy-at-microscopy.com} ; Fri, 7 Apr 2006 05:26:35 -0500 1, 21 -- Received: from lenzie.cent.gla.ac.uk ([130.209.16.18]) 1, 21 -- by hillhead.cent.gla.ac.uk with esmtp (Exim 4.10) 1, 21 -- id 1FRoAk-0000tA-00 1, 21 -- for microscopy-at-microscopy.com; Fri, 07 Apr 2006 11:26:34 +0100 1, 21 -- Received: from salt.ibls.gla.ac.uk ([130.209.54.27] helo=gould1) 1, 21 -- by lenzie.cent.gla.ac.uk with smtp (Exim 4.50) 1, 21 -- id 1FRoAk-00020g-BR 1, 21 -- for microscopy-at-microscopy.com; Fri, 07 Apr 2006 11:26:34 +0100 1, 21 -- Message-Id: {3.0.1.32.20060407112633.0089de10-at-udcf.gla.ac.uk} 1, 21 -- X-Sender: mq7x-at-udcf.gla.ac.uk 1, 21 -- X-Mailer: Windows Eudora Light Version 3.0.1 (32) 1, 21 -- Date: Fri, 07 Apr 2006 11:26:33 +0100 1, 21 -- To: microscopy-at-microscopy.com 1, 21 -- From: Martyn Quinn {mq7x-at-udcf.gla.ac.uk} 1, 21 -- Subject: Substitute for poly-L-lysine in TEM? 1, 21 -- Mime-Version: 1.0 1, 21 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
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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 4ƒC 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 NanogoldÆ 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, Jørgen.
= = = = = = = = = = = = = = = = =
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, Jørgen. } } = = = = = = = = = = = = = = = = = } } 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 £27,584 - £32,122 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, } } StÈphane } } } __________________________________________________ } 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 (£600) and the Cheaper sister the Epson V700 (£400). The older model Epson 4990 Photo is not quite as good but cheaper at £280. 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 £3k+). They all deal with scanned tif images (and much more).
However if you don't have £3,000 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, } } } } } } StÈphane } } }
==============================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==============================
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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 £250 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 £4,000.
And I remember paying £5,000 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 £10,000, and most lab mainstays like our three confocal microscopes come in at nearer £200,000 each (and users still complain about image quality).
Cheap microscopes under £100 are always a disappointment and toy (often called student) microscopes can be very poor. 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 £500 each for a quality setup (although with those like bottom end Meade [www.meade.com] at around £100 you can see something at low mag (~20x objective i.e. around 180x mag) with a quality stained section. 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 £1 and a good low mag Osram one that includes an illuminating halogen bulb at £8.
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 £60 upwards) that will be good for looking at soft static things: leaves, fruit, nuts, household objects (scan at max resolution and try reflected and 'film' mode). In the UK there are sites like http://www.wedgwood-group.com/microscopes_digital.htm that cater for schools and colleges, providing standard compound and stereo microscopes as well as cheap PC video based microscope solutions where the whole class 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 (£70) - it's fun but pretty useless for serious microscope work as it's so low res (but at 640 x 480 better than the old Intel QX-3 it replaced). It puts the image on a PC screen. 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 £500 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 £200 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%’ÑÝ } } 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, 156ñ163 (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 £1 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 DÈveloppement sur les Aliments 3600 Boul. Casavant Ouest, St-Hyacinthe, (QuÈbec) Canada J2S 8E3 TÈlÈphone/Phone: 450-773-1105 TÈlÈcopieur/ 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] EnvoyÈÝ: jeudi 29 juin 2006 21:07 ¿Ý: 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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We have been doing laser microdissection with MMI's (Molecular Machines & Industries) setup. We have been asked to do a sample for RNase analysis. MMI supplies RNase free membrane slides for the sections, but the collection tubes are not RNase free. Does anyone have a source for RNase free collection tubes for microdissection?
Mannie Steglich U T M D Anderson Cancer Center
==============================Original Headers============================== 2, 15 -- From msteglic-at-mdanderson.org Tue Jul 11 09:46:03 2006 2, 15 -- Received: from UTM-MAIL04A.mdacc.tmc.edu (utmlnmail04nt.mdacc.tmc.edu [143.111.84.152]) 2, 15 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6BEk22T028914 2, 15 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 11 Jul 2006 09:46:03 -0500 2, 15 -- To: Microscopy-at-MSA.Microscopy.Com 2, 15 -- Subject: RNase free Microdissection 2, 15 -- MIME-Version: 1.0 2, 15 -- X-Mailer: Lotus Notes Release 5.0.11 July 24, 2002 2, 15 -- Message-ID: {OF6E4AD7C7.00F49114-ON862571A8.0050B5F4-862571A8.00513D27-at-mdacc.tmc.edu} 2, 15 -- From: msteglic-at-mdanderson.org 2, 15 -- Date: Tue, 11 Jul 2006 09:46:01 -0500 2, 15 -- X-MIMETrack: Serialize by Router on UTM-MAIL04A/HOU/UTMDACC(Release 5.0.11 |July 24, 2002) at 2, 15 -- 07/11/2006 09:46:03 AM, 2, 15 -- Serialize complete at 07/11/2006 09:46:03 AM 2, 15 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
Geoff- You are correct. The Sapphatomes were sapphire knives. They were supposed to be low-cost alternatives to diamonds. They cut well, but were almost impossible to keep clean, and did not wear nearly as well as hoped. Ages ago, we had one in the lab I was in at the time. It became the training knife for a new technician. Then it went into a drawer, never again to see the light of day. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 1, 24 -- From lcgould-at-med.cornell.edu Tue Jul 11 14:28:57 2006 1, 24 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 1, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6BJSuU5021477 1, 24 -- for {microscopy-at-microscopy.com} ; Tue, 11 Jul 2006 14:28:56 -0500 1, 24 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 24 -- by smtp-gw1.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k6BJSrNO026917 1, 24 -- for {microscopy-at-microscopy.com} ; Tue, 11 Jul 2006 15:28:54 -0400 (EDT) 1, 24 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 24 -- by mpx1.med.cornell.edu 1, 24 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 24 -- with ESMTPA id {0J29002Z07G5PL50-at-mpx1.med.cornell.edu} for 1, 24 -- microscopy-at-microscopy.com; Tue, 11 Jul 2006 15:28:53 -0400 (EDT) 1, 24 -- Date: Tue, 11 Jul 2006 15:20:58 -0400 1, 24 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 24 -- Subject: Re: [Microscopy] microtome knife question 1, 24 -- In-reply-to: {200607111913.k6BJDd9P012443-at-ns.microscopy.com} 1, 24 -- Sender: lcgould-at-med.cornell.edu 1, 24 -- To: Geoffrey_Williams-at-brown.edu, 1, 24 -- Microscopy Listserver {microscopy-at-microscopy.com} 1, 24 -- Message-id: {p06230914c0d9a9a1dfb4-at-[140.251.48.23]} 1, 24 -- MIME-version: 1.0 1, 24 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 24 -- References: {200607111913.k6BJDd9P012443-at-ns.microscopy.com} 1, 24 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.7.11.121432 ==============================End of - Headers==============================
You are cordially invited to a two day, hands-on, High Pressure Freezing workshop on Aug 3 & 4, co-hosted by The Department of Molecular Genetics and Cell Biology and The Bioscience Division at The University of Chicago. The workshop will be conducted in the laboratory of Dr. Joe Austin utilizing his Bal-Tec HPM 010 High Pressure Freezer and related cryo EM sample preparation equipment. Lectures Thursday morning: 09:30 - 11:30 (Dr. Andres Kaech, Life Science Application Manager, Bal-Tec AG and Dr. Joe Austin) followed by lunch and hands-on practical lab work and instruction until 5:00pm The workshop practical sessions will continue Friday morning from 09:00 until noon Lecture room space is limited to approximately 30 people and the hands-on practical work group is restricted to a maximum of 8 participants so we encourage you to sign up early and reserve your spot. There is no charge to attend the workshop and Thursday's lunch and the Friday morning refreshments are provided with the compliments of Boeckeler Instruments.
Please email Cheryl Johnson (Cheryl-at-boeckeler.com) or Dave Roberts (dave-at-boeckeler.com) for additional details and to reserve your place.
If you are attending the Microscopy & Microanalysis meeting the University of Chicago is a 30 minute ride by cab or car from Navy Pier.
Sincerely
Dave Roberts Bal-Tec RMC Boeckeler Instruments Inc 4650 S. Butterfield Drive Tucson, AZ 85714 Tel: 520-745-0001 Fax: 520-745-0004 www.baltec-rmc.com
==============================Original Headers============================== 12, 20 -- From dave-at-boeckeler.com Tue Jul 11 17:14:34 2006 12, 20 -- Received: from txslsmtp2.vzwmail.net (txslsmtp2.vzwmail.net [66.174.85.156] (may be forged)) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6BMEYdN001717 12, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Jul 2006 17:14:34 -0500 12, 20 -- Received: from tweedledee (smtp.vzwmail.net [66.174.85.25]) 12, 20 -- (authenticated bits=0) 12, 20 -- by txslsmtp2.vzwmail.net (8.12.9/8.12.9) with ESMTP id k6BMEQgj002682 12, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 11 Jul 2006 22:14:29 GMT 12, 20 -- Message-Id: {200607112214.k6BMEQgj002682-at-txslsmtp2.vzwmail.net} 12, 20 -- From: "Dave Roberts" {dave-at-boeckeler.com} 12, 20 -- To: {microscopy-at-msa.microscopy.com} 12, 20 -- Subject: High Pressure Freezing Workshop 12, 20 -- Date: Tue, 11 Jul 2006 15:14:22 -0700 12, 20 -- MIME-Version: 1.0 12, 20 -- Content-Type: text/plain; 12, 20 -- charset="us-ascii" 12, 20 -- Content-Transfer-Encoding: 7bit 12, 20 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 12, 20 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1409 12, 20 -- Thread-Index: Acak/GnWD2jcXp0qRbaI1+GPf3gr7A== ==============================End of - Headers==============================
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Email: mckee-at-helix.mgh.harvard.edu Name: Mary McKee
Organization: MGH
Title-Subject: [Filtered] decalcification of mouse nasal bones
Question: We need to decalcify adult mouse nasal tissue for parafin embedding and possibly also for epon embedding. Does anyone have protocols for this? Thanks in advance.
Most general suppliers of labware and consumables have RNase/DNase siliconized tubes of varying sizes, ranging from 0.2 to 2.0ml. Just call your local sales rep, or go to the molecular biology people and they will be able to give you the information. In fact, they will probably just give you the tubes, they cost less than 10¢ Cdn/tube for most makes. Some expensive tubes may run up to about 15¢ Cdn each.
Alternatively, pretty well every sterile, individually wrapped polypropylene tube with cap - 6ml, is RNase/DNase free even if they are not certified.
The other point is how you are extracting the nucleic acids. If they are using guanidinium extraction then the chaotropic salts are extremely dissociative, and I have yet to see an RNase survive the extraction process. If you put the tissue directly into the Extraction Buffer system you will not have to worry about RNase/DNase until the elution stage. And I add RNasin to that stage to prevent activity then. Check with the supplier of the extraction kit you will use, their sales reps will give you fairly good advice as to the extraction process. While I have preferences, it probably does not make much difference whether you use Invitrogen, Qiagen or Roche (I apologize to those whom I am forgetting just now) or whether it is spin column or magna-bead. They all work.
Paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Bldg 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone: 204-789-3313 Pager: 204-931-9354 Cell: 204-781-1502 Fax: 204-789-3926
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Email: goldiyadav-at-yahoo.com Name: Vikas Yadav
Organization: JNU
Title-Subject: [Filtered] Golgi Staining: Rapid Golgi method of Stenasus
Question: I am trying to stain dendritic spines of rat brain using the rapid golgi method. Below are the details of the protocol i use:
RAPID GOLGI
1) 5mm slices of brain 2) Fixing solution (4 days) K2Cr2O7 ñ 5g Chloral hydrate- 5g Formaldehyde- 6ml Glutaraldehyde ñ 5ml DMSO- 5 drops Distilled H2O- 100 ml Change the solution every 2 days for 4 days. 3) Wash in 0.75%AgNO3(4 times) 4) Keep in 0.75% AgNO3- 4 days 5) Dehydration- 50%, 70%, 90% (2 hours each) 100% 2 changes of 1 hour each. 6) Ethyl Alcohol: Diethyl ether 1:1 ratio 1 hour 7) Embedding in celloidin (hot celloidin method) 8) Block (30% celloidin).
My problem is that while tissue fixation, embedding and sectioning seem to be OK the stain has apparently not penetrated deep into the tissue. While the cortical cells are well stained the deeper cells are not as well stained.
Secondly after 10 days the intensity of the stain seems to have gone down.
What could be the cause of this problewm? Can anyone help me?
I have so far been able to avoid having to section cells on their substrate (I usually pop them off) for TEM, but a client wants to look at the interaction of biofilms with the substrate. I'm looking for an opinion from anyone who has tried to thin section films on Thermanox, Permanox, or has cast their own resin slides (I think I've seen a mold for that). How likely is it that any of these materials will delaminate after embedding in epoxy resin? Acrylic resin? Slide shapes are preferable to coverslip types.
Any advice appreciated!
Mahalo, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
==============================Original Headers============================== 7, 19 -- From tina-at-pbrc.hawaii.edu Wed Jul 12 14:59:40 2006 7, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6CJxdQC032019 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 12 Jul 2006 14:59:40 -0500 7, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 7, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k6CJxbve026319 7, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 12 Jul 2006 09:59:38 -1000 (HST) 7, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k6CJxaEU026315 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 12 Jul 2006 09:59:37 -1000 (HST) 7, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 7, 19 -- Date: Wed, 12 Jul 2006 09:59:36 -1000 (HST) 7, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 7, 19 -- X-Sender: tina-at-halia 7, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 7, 19 -- Subject: Sectioning Thermanox, Permanox, resin 7, 19 -- Message-ID: {Pine.GSO.4.21.0607120935370.26169-100000-at-halia} 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
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Sorry for any delay in my reply. Overseas and having trouble sending mail!!??
Well I have kept out of the gold coated cathode debate for those that know me will already have guessed my reply!
However, I too know of the RCA gold plated cathode and 100% agree with John Bozzola in that I believe sputtering of gold would not be efficient in this case.
The answer my clients would expect is "why go to such lengths when cleaning a cathode assembly should be simple?" This is what we use -
"The cathode assembly is placed, aperture face upwards, in a beaker of stock ammonia solution diluted 3 parts ammonia to one part water. The stock solution is thought to be about 40% ammonia. After 15 minutes in the ultrasonic cleaner the beaker is placed under running water and thoroughly flushed through. Care is taken to ensure that none of the clamping or alignment screws had fallen out of the cathode assembly and could be flushed away! The cathode is then washed with alcohol before being dried with a hair drier. A new filament is fitted and centred. The assembly is checked for cleanliness by observing with a 20X lens prior to re installation in the microscope. Total time for this procedure should be less than 25 minutes break to pump down."
If the contamination in the nose of the cathode is obstinate use a little metal polish but only a polish soluble in ammonia. A follow the above procedure again but for only two or three minutes ammonia solution agitation.
The full story is available on our web site.
Steve Chapman Protrain for EM training & consultancy world wide www.emcourses.com Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967
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Email: moxleyma-at-slu.edu Name: Michael A. Moxley
Question: The correction collar on an olympus objective is very stiif almost impossible to turn. This is an inheirited scope so before investing in repairs is there anything I can do about this. Thanks
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Email: dmcdaniel-at-usuhs.mil Name: Dennis McDaniel
Organization: Uniformed Services University of the Health Sciences
I have many years experience with various types of microscopy but am relatively new to immunoEM. I am looking for a good starting protocol for gold labeling surface antigens on bacterial spore surfaces. Any nudge in the right direction would be greatly appreciated. Thanks.
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Email: m_jarnik-at-fccc.edu Name: M. Jarnik
Organization: Fox Chase Cancer Ctr
Title-Subject: [Filtered] ReAsH labeling for EM
Question: Does anybody on the list have hands-on experience with this reagent? I would like to use it both for flurescent microscopy and EM (after DAB photoconversion). A reference to a technically oriented publication would be appreciated as well.
There are many publications about labelling of suface antigens using large or small sized gold particles, pre- and postembedding.
A pre-embedding approach may be the easiest way to start, although LPS may cause steric hindrance and mask antigens in intact cells. in such cases cryoultramicrotomy may be needed to expose antigens. See e.g.
Tommassen J, Leunissen J, van Damme-Jongsten M, Overduin P. Failure of E. coli K-12 to transport PhoE-LacZ hybrid proteins out of the cytoplasm. EMBO J. 1985 Apr;4(4):1041–1047 Bosch D, Leunissen J, Verbakel J, de Jong M, van Erp H, Tommassen J. Periplasmic accumulation of truncated forms of outer-membrane PhoE protein of Escherichia coli K-12. J Mol Biol. 1986 Jun 5;189(3):449–455
Feel free to get in touch off-line for details, if you like.
Jan Leunissen
Aurion - President Research Advisor EM Costerweg 5 Dept Anatomy and Structural Biology 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4797301 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz
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I have no experience with LR-White. Yesterday I got a few ml of LR-white to try flat embedding in a petri-dish. I wanted to try chemical curing (with the accelerator) overnight at 4°C. Today the resin is still liquid and the plastic petri dish looks somewhat attacks by the product. Can't I use LR-White in plastic petri dishes (I used only ethanol)? I don't know how old is this resin, it was stored at 4°C.
I want to avoid curing in the oven, but if I cannot avoid it, I want the lowest temperature possible. I wondered if I couldn't polymerize the resin over the weekend at 40°C or 50°C.
Has someone a protocol using LR-White which would fit my wishes?
Regards,
Stephane
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Flat embedding LRW is always fun. The key is that, since it won't polymerize in the presence of oxygen, the molds holding sample and resin must be protected from oxygen by some means. We often use Thermonox cover slips (large rectangular ones) to cover the mold openings, because it is easy to peel them off after polymerization without the breakage of glass coverslips.. It is necessary to fill adjacent molds with LRW as "buffers" and cover them too, since the molds near the edges of the coverslips tend to lose lots of resin due to evaporation. This unit can be heat-polymerized like other resins or UV-polymerized in an ice chest or refrigerator to keep temperatures low. We often use molds from Ted Pella (cat. No. 10506, for example), for this purpose since they are strips held flat by a metal frame and exactly fit 22x60 mm Thermonox cover slips to seal the openings. We overfill the openings with resin so that there is overflow and the coverslips seal well. Other suppliers may have similar molds or others that would work as well.
Other options would be polymerizing in a sealed container filled with nitrogen or argon or some other gas that is not oxygen, or polymerizing in a vacuum chamber. My one experience with the latter resulted in the loss of nearly all the LRW in the mold as it evaporated, but it should be relatively easy to do some variation on the coverslip procedure to keep the resin from sublimely wandering off.
Hope this helps. Good luck.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Friday, July 14, 2006 1:24 AM To: Tindall, Randy D.
Dear listers,
I have no experience with LR-White. Yesterday I got a few ml of LR-white to try flat embedding in a petri-dish. I wanted to try chemical curing (with the accelerator) overnight at 4°C. Today the resin is still liquid and the plastic petri dish looks somewhat attacks by the product. Can't I use LR-White in plastic petri dishes (I used only ethanol)? I don't know how old is this resin, it was stored at 4°C.
I want to avoid curing in the oven, but if I cannot avoid it, I want the lowest temperature possible. I wondered if I couldn't polymerize the resin over the weekend at 40°C or 50°C.
Has someone a protocol using LR-White which would fit my wishes?
Regards,
Stephane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
When using flat embedding molds with resins that creep, we found that if you put a flat-ended cryo pin into the resin, they don't creep. This works in an AFS chamber where the atmosphere is nitrogen. Elaine
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-- Dr. Elaine Humphrey Microscopy Specialist President, Microscopy Society of Canada (2003-2005) University of British Columbia e-mail: ech-at-interchange.ubc.ca
POSTDOCTORAL POSITION AVAILABLE IMMEDIATELY DEPARTMENT OF BIOCHEMISTRY AND BIOPHYSICS UNIVERSITY OF CALIFORNIA AT SAN FRANCISCO, CALIFORNIA, USA
A post-doctoral position is available immediately in the laboratory of Gang Ren at University of California at San Francisco (UCSF). The qualified candidate may participate in ongoing research projects of the Ren laboratory. The successful candidate should have a Ph.D. degree with highly motivation and interest in structure biology. Experiences in electron microscopy, computer processing/programming, crystallography and/or protein expression and purification will be advantaged.
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Interested candidates should send a CV including areas of expertise and research interest, publications list, and names and contact information for 3 references to Dr. Gang Ren at gren-at-msg.ucsf.edu.
Gang ‘Gary’ Ren, Ph.D. University of California, San Francisco Department of Biochemistry & Biophysics San Francisco, CA 94158 Email: gren-at-msg.ucsf.edu
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This Question was submitted to Ask-A-Microscopist by (cartoonish2-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, July 15, 2006 at 06:58:20 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both cartoonish2-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: cartoonish2-at-yahoo.com Name: nish pandya
Education: Graduate College
Location: washington dc usa
Title: career
Question: Hi.
Wanted to get your advice on how I can become a Microscopist. I have a Master's degree in biomedical engineering from the Univ of Miami. My graduate work was in biomedical optics and laser applications. I also did some research at Duke Univ. For the past 4 years, I have worked as a Software Engineer but I want to get back into the field of optics. Do you have any suggestions on how to get into the field ? I am willing to further my education. I just don't know where to look for resources. Thanks.
Question: Once again the MSA Education Committee is organizing Exhibitor Demonstrations and Tutorials at the MM2006 Meeting. These tutorials will take place on Tuesday, August 1 starting at 5:00 pm in the Navy Pier Exhibit Hall. These mini-seminars or tutorial demonstrations are held in the booths of the participating companies after the xhibit Hall is closed to non-participants.
Reservations sheets with titles and descriptions will be at the MSA Education table in the MSA Mega Booth. When you sign up you will be issued a ticket, which you will need to renter the Hall after it is closed. You need to sign up no later than noon on Tuesday. The number of attendees is limited, so visit the MSA MegaBooth soon, since the demonstrations get filled up quickly!
You may download a PDF file with an extended outline of the participating Exhibitors as well as titles of all the presentations at http://mm2006.microscopy.org . Then follow the links to the Exhibitors or Tutorials pages.
Dr. Andreas Holzenburg MSA Education Committee - Exhibitors Tutorials SubCommittee Chair
Why would you want to do that? Any idiot with a microscope can be called a microscopist. It is just like anyone with a camera can be a landscape photographer. Semi-true but much more complicated. Been there, done that.
You can make much more income doing software work than traditional tech SEM work. Frankly, I don't see EM as a big money maker or legacy venue. You are way better off where you are. And you can grow to bigger venues.
If you add computer engineering to your CV, that will IMO greatly assist you. Stop and think about what you want to do versus what you need to do. Your personal preferences may override all other inputs. That is fine. Just be aware of the consequences and cost of each decision.
But fundamentally--why do you want to be a microscopist? TEM, SEM, STEM, AFM, etc.? You will get solicitations that expect you to have a Ph.D in something and scads of experience in all of the specific units that the solicitor has. Either this is for one specific person or for a phantom.
Sigh....the EM folks may not like this posting. But it is the truth and from the heart.
Why am I a microscopist? I love it. EM or LM. But I am also an EE. The SEM and LM are merely tools. But they are not simple to use. However, adding EE is not a simple nor easy process. You will find this to be akin to computer engineering.
gary g.
At 08:05 AM 7/15/2006, you wrote:
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I did not mean to sound harsh. If I did, I apologize for that.
What I was trying to say, obviously with a poor approach, was that to do microscopy well requires a lot of training, work and perhaps a bit of luck. But after all of the training and course work, what type of job can one find? The MRS Bulletin lists a few job offerings. Most require a Ph.D. and typically pay $60K or so. Some jobs are available as "technicians" but these usually pay less. It is interesting that the majority of microscopy job openings are published in non-microscopy materials. Zero in Microscopy Today and I do not recall seeing one in an M&M issue--well, maybe one some time ago.
I don't like low paying EM jobs. That is why I'm an EE. The EM is simply a tool. But it is a great tool. And it can do really fantastic things.
You are right about optics. He made the initial venture into microscopy but it sounds like he really wants to get into optical physics. That is different.
gary g.
At 10:51 AM 7/16/2006, you wrote: } Hello Gary: } } I think you are very harsh to Nish. I am not sure you gave him/her the } proper advice sought. If you love microscopy with its low pay, someone } else could, don't you think so? } } Nish can get into microscopy (actually, optics, as he/she seems to } suggest) via a formal training in materials science/solid state } physics. Since he/she already has a degree in an engineering } discipline, he/she would naturally fit into materials/solid state } physics. What if Nish is intersted in synchrotron science? There are } several "optics" thing in that area too. } } Thanks and have a nice day. } } Ike Oguocha } --- gary-at-gaugler.com wrote: } } } Why would you want to do that? Any idiot with a } } microscope can be called a microscopist. It is just } } like anyone with a camera can be a landscape photographer. } } Semi-true but much more complicated. Been there, done that. } } } } You can make much more income doing software } } work than traditional tech SEM work. Frankly, } } I don't see EM as a big money maker or legacy } } venue. You are way better off where you are. } } And you can grow to bigger venues. } } } } If you add computer engineering to your CV, that } } will IMO greatly assist you. Stop and think about } } what you want to do versus what you need to do. } } Your personal preferences may override all other } } inputs. That is fine. Just be aware of the consequences } } and cost of each decision. } } } } But fundamentally--why do you want to be a microscopist? } } TEM, SEM, STEM, AFM, etc.? You will get solicitations that } } expect you to have a Ph.D in something and scads of } } experience in all of the specific units that the solicitor } } has. Either this is for one specific person or for a phantom. } } } } Sigh....the EM folks may not like this posting. But it is } } the truth and from the heart. } } } } Why am I a microscopist? I love it. EM or LM. But I } } am also an EE. The SEM and LM are merely tools. But they } } are not simple to use. However, adding EE is not a simple nor } } easy process. You will find this to be akin to computer } } engineering. } } } } gary g. } } } } } } At 08:05 AM 7/15/2006, you wrote: } } } } } } } } } on Saturday, July 15, 2006 at 06:58:20 } } } Remember to consider the Grade/Age of the student when considering } } } the Question } } } } } } Email: cartoonish2-at-yahoo.com } } } Name: nish pandya } } } } } } Education: Graduate College } } } } } } Location: washington dc usa } } } } } } Title: career } } } } } } Question: Hi. } } } } } } Wanted to get your advice on how I can become a Microscopist. I } } } have a Master's degree in biomedical engineering from the Univ of } } } Miami. My graduate work was in biomedical optics and laser } } } applications. I also did some research at Duke Univ. } } } For the past 4 years, I have worked as a Software Engineer but I } } } want to get back into the field of optics. } } } Do you have any suggestions on how to get into the field ? I am } } } willing to further my education. I just don't know where to look } } } for resources. } } } Thanks. } } } } } } } } } } ___________________________________________________________ } All new Yahoo! Mail "The new Interface is stunning in its simplicity } and ease of use." - PC Magazine } http://uk.docs.yahoo.com/nowyoucan.html
==============================Original Headers============================== 8, 21 -- From gary-at-gaugler.com Sun Jul 16 13:21:56 2006 8, 21 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k6GILtrb015074 8, 21 -- for {microscopy-at-microscopy.com} ; Sun, 16 Jul 2006 13:21:56 -0500 8, 21 -- Received: (qmail 1770 invoked from network); 16 Jul 2006 11:21:54 -0700 8, 21 -- Received: by simscan 1.1.0 ppid: 1767, pid: 1768, t: 0.1830s 8, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 8, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 8, 21 -- by qsmtp2 with SMTP; 16 Jul 2006 11:21:54 -0700 8, 21 -- Message-Id: {7.0.1.0.2.20060716110359.02528658-at-gaugler.com} 8, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 8, 21 -- Date: Sun, 16 Jul 2006 11:21:55 -0700 8, 21 -- To: Ike Oguocha {oguocha-at-yahoo.com} 8, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 8, 21 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: how I can become a 8, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} , cartoonish2-at-yahoo.com 8, 21 -- In-Reply-To: {20060716175114.49347.qmail-at-web53715.mail.yahoo.com} 8, 21 -- References: {200607160618.k6G6ID7P015936-at-ns.microscopy.com} 8, 21 -- {20060716175114.49347.qmail-at-web53715.mail.yahoo.com} 8, 21 -- Mime-Version: 1.0 8, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Nish Pandya wrote: =========================================================== Wanted to get your advice on how I can become a Microscopist. I have a Master's degree in biomedical engineering from the Univ of Miami. My graduate work was in biomedical optics and laser applications. I also did some research at Duke Univ. For the past 4 years, I have worked as a Software Engineer but I want to get back into the field of optics. Do you have any suggestions on how to get into the field ? I am willing to further my education. I just don't know where to look for resources. Thanks. ========================================================== I am not sure there is any "one" best way that one becomes a microscopist. I am sure that we could even have a lively dialogue as to just what constitutes being a "microscopist". But there are numerous ways one could "become" a microscopist, from enrolling in courses to taking a position in a setting where you would be working under an experienced microscopist for a period of time.
But I would like to comment on previous comments to your question about opportunities in microscopy. From my vantage point, the opportunities have never appeared greater in the nearly 40 years since leaving graduate school myself. Certainly in the USA, there has been a huge proliferation of new start-up companies in areas such as nanotechnology and biomaterials, and one just does not get very far in those areas without having on board an experienced microscopy department and laboratory. There is also, again, at least in the USA, a growth in the number of junior colleges and even high schools that are putting in SEMs for the benefit of their students, and of course, one knowledgeable about microscopy is needed for those institutional situations as well. Anyone attending several recent exhibitions of the MSA (e.g. the "M&M" meeting) would also see that there is an ever increasing number of firms offering microscopy-related products, and with increasingly greater sophistication and technology, and such firms could not operate without employing an increasingly larger number of experienced microscopists.
The issue of salary is indeed important and I see our industry as being very very healthy and it would not be attracting the good people it needs to grow and innovate if it was not paying the kinds of salaries that could attract the best of minds. Certainly there is a spread in terms of what people earn, just as there is for just about any other walk of life. But I would not agree with the suggestion that the opportunities, professional or financial, are not present for those contemplating a career in microscopy.
Chuck
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Obviously the plastic container was the cause of the bad polymerization since curing in Falcon tubes or eppendorfs works and the blocks are ready in 20 min with the accelerator. I am currently trying to find the ideal container for my purpose but it is really not easy. It must be in PE or PP, have a flat bottom and be large enough to accomodate 18x18 coverslips and it must be one-use.
Chemical curing is really easier than the other methods because one does not need to care about the presence of air and it is much (and I mean much) faster. Notwithstanding I wonder how it is possible to heat in oven AND at the same time keep a nitrogen atmosphere. Why couldn't one simply cover the resin with oil to avoid contact with air? Or another liquid which does not mixes with the resin?
BTW, can someone tell me what plastic are made the caps of 50 ml falcon tubes? I couldn't find this information.
regards,
Stéphane
--- nizets2-at-yahoo.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear listers, } } I have no experience with LR-White. Yesterday I got } a } few ml of LR-white to try flat embedding in a } petri-dish. I wanted to try chemical curing (with } the } accelerator) overnight at 4°C. Today the resin is } still liquid and the plastic petri dish looks } somewhat } attacks by the product. Can't I use LR-White in } plastic petri dishes (I used only ethanol)? I don't } know how old is this resin, it was stored at 4°C. } } I want to avoid curing in the oven, but if I cannot } avoid it, I want the lowest temperature possible. I } wondered if I couldn't polymerize the resin over the } weekend at 40°C or 50°C. } } Has someone a protocol using LR-White which would } fit } my wishes? } } Regards, } } Stephane } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam } protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 7, 18 -- From nizets2-at-yahoo.com Fri Jul 14 01:22:05 } 2006 } 7, 18 -- Received: from web37414.mail.mud.yahoo.com } (web37414.mail.mud.yahoo.com [209.191.87.67]) } 7, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } k6E6M5XL016952 } 7, 18 -- for {microscopy-at-microscopy.com} ; Fri, 14 } Jul 2006 01:22:05 -0500 } 7, 18 -- Received: (qmail 88984 invoked by uid } 60001); 14 Jul 2006 06:22:04 -0000 } 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; } c=nofws; } 7, 18 -- s=s1024; d=yahoo.com; } 7, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 7, 18 -- } b=iz7y9kHVNTA7vVqeF0/eP8sTXePnPx2ykJjmp547BWX3234uTXAOwrmqeVZ1PhYMSnv3A28RmFE3xzXMxuWyjQ0aoq8Ts3j87PpI2B87dOLqPrC1jyPDlEvomV5DpRbpma7L0g9GrXaMHSse319FTP3AwxvFvi0TEavGtOqyQys= } ; } 7, 18 -- Message-ID: } {20060714062204.88982.qmail-at-web37414.mail.mud.yahoo.com} } 7, 18 -- Received: from [80.122.101.102] by } web37414.mail.mud.yahoo.com via HTTP; Thu, 13 Jul } 2006 23:22:04 PDT } 7, 18 -- Date: Thu, 13 Jul 2006 23:22:04 -0700 (PDT) } 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 7, 18 -- Subject: Urgent HELP: LR-White informations } needed } 7, 18 -- To: microscopy-at-microscopy.com } 7, 18 -- MIME-Version: 1.0 } 7, 18 -- Content-Type: text/plain; } charset=iso-8859-1 } 7, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== }
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
I'm a generalist. I would not want it any other way. I use PLM, phase contrast, dispersion staining, SEM, EDS, TEM and microchemical testing. I've polished thin sections of minerals, coal and pencil urchin spines. I made blood spears and cut thin sections of wood, rubber and plastics. I've polished metal and cross sections of dipped kevlar cord. I can't imaging spending 30 years doing citric acid titrations or measuring beach sand particles.
Do I have a career? No, I have an advocation. Am I rich? No, but I've made a good living doing what I would do as a hobby if I could afford the equipment.
You want to be a microscopist? Blow off all of this advise and study microscopy. Learn about PLN and image analysis. Study SEM and TEM/EELS. Learn to take photomicrographs that people want to see. Poke around in microchemical testing and staining. Look at some minerals from the point of view as a geologist and then as the forensic microscopist. Associate with other microscopists. And when you find an area you like stay with it.
My first job out of college was a QC chemist. I hated it. I started sleeping later and later every day. I started at work arriving later and later every day. Fortunately my interest in microscopy found me a position I wanted. I like being a generalist. Yes I look forward to Friday night, but come Sunday night, I start thinking about the job and most of the time I'm fired up about starting a new work week. I know people who make a lot more money, but they also hate their job.
Now I gotta go... I have about 500 particle of carbon black i have to measure and i can't wait to find out if they'll match the earlier data!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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==============================Original Headers============================== 12, 18 -- From frank.karl-at-degussa.com Mon Jul 17 08:03:16 2006 12, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 12, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HD3FCL018106 12, 18 -- for {microscopy-at-msa.microscopy.com} ; Mon, 17 Jul 2006 08:03:15 -0500 12, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 12, 18 -- by framailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k6HD3BAQ030210 12, 18 -- for {microscopy-at-msa.microscopy.com} ; Mon, 17 Jul 2006 15:03:12 +0200 12, 18 -- In-Reply-To: {200607170419.k6H4JEPW025439-at-ns.microscopy.com} 12, 18 -- Subject: Re: [Microscopy] More on "becoming a microscopist" 12, 18 -- To: microscopy-at-msa.microscopy.com 12, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 12, 18 -- Message-ID: {OF7C0E96DB.EA10FE62-ON852571AE.0045E79E-852571AE.0047B1FC-at-degussa.com} 12, 18 -- From: frank.karl-at-degussa.com 12, 18 -- Date: Mon, 17 Jul 2006 09:03:06 -0400 12, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 12, 18 -- 07/17/2006 08:03:13 AM 12, 18 -- MIME-Version: 1.0 12, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Yep, flat embedding with LR-White has some problems. You are correct =20=
that the plastic dish is a problem. Oxygen is also a problem. Here =20 is what I have done.
Coat TC plates with sterile, molten 1.5% Agar, 0.5% Gelatin in =20 ddH2O. Wash around the inside of the plate and on the top of the =20 plate cover, pour out the excess and let dry. Set up cells. Cells may not adhere to this coating as well as to =20 plastic, so you may need to increase the length of cell binding =20 incubations (and be more gentle with future treatments). Do all of the things you normally do. Change the LR-White several =20 times. The final should fill the TC dish to over full. Place the =20 cover upside-down on top of the Dish (diagonally) to exclude all air.
The covering of the Agar & Gelatin will keep the LR-White from =20 interacting with the plastic of the dish. Having the dish full of LR- =20=
White and putting the cover on the dish (Agar/Gelatin side in contact =20=
with the LR-White) will exclude most of the oxygen. This has worked =20 for me in the past. Let me know if you have any questions.
David
On Jul 13, 2006, at 11:26 PM, nizets2-at-yahoo.com wrote:
--| ------ --| --| Dear listers, --| --| I have no experience with LR-White. Yesterday I got a --| few ml of LR-white to try flat embedding in a --| petri-dish. I wanted to try chemical curing (with the --| accelerator) overnight at 4=B0C. Today the resin is --| still liquid and the plastic petri dish looks somewhat --| attacks by the product. Can't I use LR-White in --| plastic petri dishes (I used only ethanol)? I don't --| know how old is this resin, it was stored at 4=B0C. --| --| I want to avoid curing in the oven, but if I cannot --| avoid it, I want the lowest temperature possible. I --| wondered if I couldn't polymerize the resin over the --| weekend at 40=B0C or 50=B0C. --| --| Has someone a protocol using LR-White which would fit --| my wishes? --| --| Regards, --| --| Stephane
==============================Original Headers============================== 13, 20 -- From Elliott-at-arizona.edu Mon Jul 17 09:19:41 2006 13, 20 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HEJfou029415 13, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 17 Jul 2006 09:19:41 -0500 13, 20 -- Received: from localhost (eomer.email.arizona.edu [10.0.0.219]) 13, 20 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 32944EA98BB 13, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 17 Jul 2006 07:19:41 -0700 (MST) 13, 20 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 13, 20 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id E637EEA9624 13, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 17 Jul 2006 07:19:39 -0700 (MST) 13, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 13, 20 -- Content-Transfer-Encoding: 7bit 13, 20 -- Message-Id: {C46C7CD7-5FB2-423F-B14C-4A426291871E-at-arizona.edu} 13, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 13, 20 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 13, 20 -- From: David Elliott {Elliott-at-arizona.edu} 13, 20 -- Subject: Re: LR-White information needed 13, 20 -- Date: Mon, 17 Jul 2006 07:19:38 -0700 13, 20 -- X-Mailer: Apple Mail (2.752.2) 13, 20 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Hi Listers, Anyone out there have a set of operator's manuals (I have the schematics, so I won't need these) for the Stereoscan 260 SEM that I may copy or purchase? Please contact me off list, thanks.
Gary Easton
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==============================Original Headers============================== 7, 20 -- From garyeaston-at-scannerscorp.com Mon Jul 17 09:27:11 2006 7, 20 -- Received: from omr1.networksolutionsemail.com (omr1.networksolutionsemail.com [205.178.146.51]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HERAdh007174 7, 20 -- for {microscopy-at-microscopy.com} ; Mon, 17 Jul 2006 09:27:10 -0500 7, 20 -- Received: from mail.networksolutionsemail.com (ns-omr1.mgt.hosting.dc2.netsol.com [10.49.6.64]) 7, 20 -- by omr1.networksolutionsemail.com (8.13.6/8.13.6) with SMTP id k6HER8JL019862 7, 20 -- for {microscopy-at-microscopy.com} ; Mon, 17 Jul 2006 10:27:08 -0400 7, 20 -- Received: (qmail 24039 invoked by uid 78); 17 Jul 2006 14:26:48 -0000 7, 20 -- Received: from unknown (HELO ?192.168.1.46?) (70.22.76.210) 7, 20 -- by 10.49.36.64 with SMTP; 17 Jul 2006 14:26:48 -0000 7, 20 -- Received: from 127.0.0.1 (AVG SMTP 7.1.394 [268.10.1/389]); Mon, 17 Jul 2006 10:26:41 -0400 7, 20 -- Message-ID: {44BB9E21.5000108-at-scannerscorp.com} 7, 20 -- Date: Mon, 17 Jul 2006 10:26:41 -0400 7, 20 -- From: "Gary M. Easton" {garyeaston-at-scannerscorp.com} 7, 20 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 7, 20 -- To: Microscopy Society of America {microscopy-at-microscopy.com} 7, 20 -- Subject: CAMBRIDGE/LEICA/LEO S260 SEM MANUALS 7, 20 -- Mime-Version: 1.0 7, 20 -- Content-Transfer-Encoding: 7bit 7, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both amyhinsley-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: amyhinsley-at-yahoo.com Name: Amy
Title-Subject: [Filtered] PLM training
Question: I'm interested in attending a course for PLM for asbestos identification and so far have only been able to find the McCrone Institute. Are there other options?
This Question was submitted to Ask-A-Microscopist by (fortier-at-slu.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, July 17, 2006 at 14:24:19 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both fortier-at-slu.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: fortier-at-slu.edu Name: Joseph Fortier
Organization: Saint Louis University
Education: Graduate College
Location: Saint Louis, MO, U.S.A.
Title: critical point drying insects
Question: In order to prepare very small (less than or equal to 5mm) insects for electron microscopy, is it necessary to chemically dry them with acetone and amyl acetate in addition to CPD, if they have already been prepared in 100% ethanol?
If the samples are currently in 100% ethanol, you can go directly into the CPD device. Passage through amyl acetate is not necessary since liquid CO2 is miscible with absolute ethanol. Simply, place 5-10 ml of abs ethanol in the CPD chamber to maintain a saturated ethanol atmosphere (thereby preventing evaporation and drying of the insects), transfer the insects into the chamber and CPD as usual. Since the specimens are so small, I assume that they will be placed into appropriate carriers, otherwise they will be lost in the exchange of CO2. I do not believe that liquid CO2 will extract DNA, but I would defer to a molecular biologist if they now otherwise.
If you need more info, please do not hesitate to ask.
JB
} } Email: fortier-at-slu.edu } Name: Joseph Fortier } } Organization: Saint Louis University } } Education: Graduate College } } Location: Saint Louis, MO, U.S.A. } } Title: critical point drying insects } } Question: In order to prepare very small (less than or equal to 5mm) } insects for electron microscopy, is it necessary to chemically dry } them with acetone and amyl acetate in addition to CPD, if they have } already been prepared in 100% ethanol? } } Also, I wonder if the above chemical damage DNA? } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 9, 12 -- From zaluzec-at-microscopy.com Mon Jul 17 15:17:17 2006 } 9, 12 -- Received: from [10.255.254.48] (msdvpn24.msd.anl.gov } [130.202.238.88]) } 9, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k6HKHFk0024720 } 9, 12 -- for {microscopy-at-microscopy.com} ; Mon, 17 Jul 2006 } 15:17:16 -0500 } 9, 12 -- Mime-Version: 1.0 } 9, 12 -- X-Sender: (Unverified) } 9, 12 -- Message-Id: {p06020402c0e1a0be0f7d-at-[10.255.254.48]} } 9, 12 -- Date: Mon, 17 Jul 2006 16:17:19 -0400 } 9, 12 -- To: microscopy-at-microscopy.com } 9, 12 -- From: fortier-at-slu.edu (by way of Ask-A-Microscopist) } 9, 12 -- Subject: AskAMicroscopist: critical point drying insects } 9, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers==============================
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
Thomas A. Kubik & Assoc. (TAKA) P.O. Box 208 Greenlawn, NY 11704 Phone: 516-261-2117
Peter M. Cooke Microscopy Instruction, Consultation & Analysis (MICA) 5807 N. Maplewood Chicago, IL 60659 Phone: 773-334-2240 pmcooke-at-earthlink.net
I do not know much about TAKA, so I can't give you any first hand advise about the company.
I can provide you some information about MICA though. Peter was an instructor at McCrone for many years before starting up his own company. I took my PLM training courses with him at McCrone and have used his company to train new PLM analysts at two laboratories which I have managed. His courses are taught both in Chicago and on-site if needed. I consider him the best asbestos PLM trainer in the USA. (I also have NO financial interest in MICA).
Good luck in whichever training course you choose.
Steven Grevelis Asbestos Laboratory Manager Groundwater Analytical, Inc. Buzzards Bay, MA 02532 Ph. 508.759.4441 Fax 508.759.4475 www.groundwateranalytical.com
-----Original Message----- X-from: amyhinsley-at-yahoo.com [mailto:amyhinsley-at-yahoo.com] Sent: Monday, July 17, 2006 4:30 PM To: sgrevelis-at-groundwateranalytical.com
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both amyhinsley-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: amyhinsley-at-yahoo.com Name: Amy
Title-Subject: [Filtered] PLM training
Question: I'm interested in attending a course for PLM for asbestos identification and so far have only been able to find the McCrone Institute. Are there other options?
Go from 100% ethanol directly to the CPD using ethanol as your transition solvent. The other solvents are not necessary.
I have also just recently run across a paper that uses the CPD to reverse the effects of chemical fixation with formalin allowing sequencing so I would surmise damage if any is negligible.
Good luck
Scott Whittaker NMNH Imaging Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-633-0891
-----Original Message----- X-from: fortier-at-slu.edu [mailto:fortier-at-slu.edu] Sent: Monday, July 17, 2006 4:21 PM To: Whittaker, Scott
This Question was submitted to Ask-A-Microscopist by (fortier-at-slu.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, July 17, 2006 at 14:24:19 Remember to consider the Grade/Age of the student when considering the Question ------------------------------------------------------------------------ --- Please reply to both fortier-at-slu.edu as well as to the Microscopy Listserver ------------------------------------------------------------------------ ---
Email: fortier-at-slu.edu Name: Joseph Fortier
Organization: Saint Louis University
Education: Graduate College
Location: Saint Louis, MO, U.S.A.
Title: critical point drying insects
Question: In order to prepare very small (less than or equal to 5mm) insects for electron microscopy, is it necessary to chemically dry them with acetone and amyl acetate in addition to CPD, if they have already been prepared in 100% ethanol?
Hello Joseph. I just returned from Woods Hole, Massachusetts where we did some SEM on some small ants, Aphids and spiders. An entomologist there gave us a protocol where you rinse the bugs in 70% ETOH and directly sputter coat. I am not sure whether the ETOH killed them completely. We carefully mounted them on double stick tape on stubs and sputter coated them. The structure was very nice and the preparation extremely easy. Good Luck JoAnn Buchanan Stanford University School of Medicine At 01:22 PM 7/17/2006, you wrote:
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==============================Original Headers============================== 6, 21 -- From redhair-at-stanford.edu Mon Jul 17 16:25:00 2006 6, 21 -- Received: from smtp3.stanford.edu (smtp3.Stanford.EDU [171.67.20.26]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HLOxp3011171 6, 21 -- for {microscopy-at-microscopy.com} ; Mon, 17 Jul 2006 16:24:59 -0500 6, 21 -- Received: from smtp3.stanford.edu (localhost.localdomain [127.0.0.1]) 6, 21 -- by localhost (Postfix) with SMTP id 4DEAD4CA81; 6, 21 -- Mon, 17 Jul 2006 14:24:55 -0700 (PDT) 6, 21 -- Received: from BIG-DADDY.stanford.edu (ssmith-pbdsl1.Stanford.EDU [171.66.197.50]) 6, 21 -- by smtp3.stanford.edu (Postfix) with ESMTP id E38504C839; 6, 21 -- Mon, 17 Jul 2006 14:24:28 -0700 (PDT) 6, 21 -- Message-Id: {6.2.5.6.2.20060717141736.03765af0-at-stanford.edu} 6, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.5.6 6, 21 -- Date: Mon, 17 Jul 2006 14:24:27 -0700 6, 21 -- To: fortier-at-slu.edu, "Microscopy Listserver" {microscopy-at-microscopy.com} 6, 21 -- From: JoAnn Buchanan {redhair-at-stanford.edu} 6, 21 -- Subject: Re: [Microscopy] AskAMicroscopist: critical point drying 6, 21 -- insects 6, 21 -- In-Reply-To: {200607172022.k6HKMslc032695-at-ns.microscopy.com} 6, 21 -- References: {200607172022.k6HKMslc032695-at-ns.microscopy.com} 6, 21 -- Mime-Version: 1.0 6, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Thomas A. Kubik & Assoc. (TAKA) P.O. Box 208 Greenlawn, NY 11704 Phone: 516-261-2117
Peter M. Cooke Microscopy Instruction, Consultation & Analysis (MICA) 5807 N. Maplewood Chicago, IL 60659 Phone: 773-334-2240 pmcooke-at-earthlink.net
I do not know much about TAKA, so I can't give you any first hand advise about the company.
I can provide you some information about MICA though. Peter was an instructor at McCrone for many years before starting up his own company. I took my PLM training courses with him at McCrone and have used his company to train new PLM analysts at two laboratories which I have managed. His courses are taught both in Chicago and on-site if needed. I consider him the best asbestos PLM trainer in the USA. (I also have NO financial interest in MICA).
Good luck in whichever training course you choose.
Steven Grevelis Asbestos Laboratory Manager Groundwater Analytical, Inc. Buzzards Bay, MA 02532 Ph. 508.759.4441 Fax 508.759.4475 www.groundwateranalytical.com
I know Peter as well as Thomas "Tom" A. Kubik & Assoc. (TAKA); I am biased though as I teach at the McCrone Research Institute.
Peter is a good teacher and has been an NVLAP inspector.
Tom is well versed in PLM and acceptable as well; been around for a long time.
There are a couple on the West Coast (Seattle and CA) that I know of but too indirectly now and thus will not commet on because of that.
Another option is the Environmental Institute in Atlanta/Marietta, GA. Dave Hogue (formerly of GA Tech RI) runs the place and Tom Laubenthal usually teaches the PLM asbestos course. Tom has been around for years (we met when we both worked at McCrone in Atlanta in 1987) and has been an NVLAP inspector. He knows the PLM, asbestos and is not too shabby on the regs as well as field work. Unfortunately, Tom has become a little too jaded over the years (like myself) and it tends to slip into the training. Would still recommend him though.
The Environmental Institute 1300 Williams Drive Marietta, GA 30066 (770) 427-3600 (770) 421-2484 Fax info-at-tei-atl.com
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
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==============================Original Headers============================== 23, 31 -- From ph2-at-sprynet.com Mon Jul 17 18:18:05 2006 23, 31 -- Received: from elasmtp-spurfowl.atl.sa.earthlink.net (elasmtp-spurfowl.atl.sa.earthlink.net [209.86.89.66]) 23, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6HNI4AY022580 23, 31 -- for {Microscopy-at-Microscopy.Com} ; Mon, 17 Jul 2006 18:18:05 -0500 23, 31 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 23, 31 -- s=dk20050327; d=sprynet.com; 23, 31 -- b=JjKbfG34jr3nM5RJe726NY/WnBuLD3DchXSqZFkI+PwaMTkKTtrKuPbNl31VMmeT; 23, 31 -- h=Received:Reply-To:From:To:Subject:Date:Organization:Message-ID:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Priority:X-MSMail-Priority:X-Mailer:X-MimeOLE:Importance:X-ELNK-Trace:X-Originating-IP; 23, 31 -- Received: from [4.224.249.111] (helo=yourw92p4bhlzg) 23, 31 -- by elasmtp-spurfowl.atl.sa.earthlink.net with asmtp (TLSv1:RC4-MD5:128) 23, 31 -- (Exim 4.34) 23, 31 -- id 1G2cLe-0004FK-Q5; Mon, 17 Jul 2006 19:18:02 -0400 23, 31 -- Reply-To: {ph2-at-sprynet.com} 23, 31 -- From: "Tony Havics" {ph2-at-sprynet.com} 23, 31 -- To: "Microscopy Listserve" {Microscopy-at-Microscopy.Com} 23, 31 -- Subject: PLM Training on Asbestos 23, 31 -- Date: Mon, 17 Jul 2006 19:24:28 -0500 23, 31 -- Organization: pH2 23, 31 -- Message-ID: {000201c6aa00$8b464d00$6ff9e004-at-QEPI.local} 23, 31 -- MIME-Version: 1.0 23, 31 -- Content-Type: text/plain; 23, 31 -- charset="utf-8" 23, 31 -- X-Priority: 3 (Normal) 23, 31 -- X-MSMail-Priority: Normal 23, 31 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 23, 31 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 23, 31 -- Importance: Normal 23, 31 -- X-ELNK-Trace: 6888e50b2be9b4fee5331016acda17f9973d681e0e4feb7e488d415b7b9318c3350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c 23, 31 -- X-Originating-IP: 4.224.249.111 23, 31 -- Content-Transfer-Encoding: 8bit 23, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6HNI4AY022580 ==============================End of - Headers==============================
Sorry I did not provide the reference. It had just hit my hand a couple weeks ago but not yet been entered into my database and my assistant wasn't around to ask where it was.
Formalin Removal from Archival Tissue by Critical Point Drying Biotechniques 33:604-611 September (2002).
Scott Whittaker NMNH Imaging Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-633-0891
-----Original Message----- X-from: Jim Ehrman [mailto:jehrman-at-mta.ca] Sent: Monday, July 17, 2006 7:17 PM To: Whittaker, Scott
Hello, I have used the microwave from Pella for TEM/SEM processing. However I don't have a budget for one right now.
I was wondering if anyone has tried using the new variable wattage microwaves available for consumer use? One I was considering testing is from Panasonic. The technology is explained here:
or a link if wrapping beyond the margins doesn't work: http://makeashorterlink.com/?M24131D6D
Granted, I may have to make use of water baths to avoid hot spots, but it is a far cheaper route than the $15k. The wattage for histology microwaves are variable from 20-1000 watts, or 250-750 watts. The full power of the panasonic one is 1250 watts, but has low; medium; and high wattage settings.
Anyone given these a try, or know of other commercial microwaves with variable wattage settings that report actual wattage numbers? Thanks for your help.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 8, 24 -- From gvrdolja-at-nature.berkeley.edu Tue Jul 18 11:35:02 2006 8, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6IGZ1eV020273 8, 24 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 11:35:02 -0500 8, 24 -- Received: from localhost (localhost [127.0.0.1]) 8, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 13872C1E69 8, 24 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 09:35:00 -0700 (PDT) 8, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 8, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 8, 24 -- with ESMTP id 26824-02 for {microscopy-at-microscopy.com} ; 8, 24 -- Tue, 18 Jul 2006 09:34:45 -0700 (PDT) 8, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 8, 24 -- id 186B2C1E59; Tue, 18 Jul 2006 09:34:45 -0700 (PDT) 8, 24 -- Received: from localhost (localhost [127.0.0.1]) 8, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 0EBD3C1E48 8, 24 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 09:34:44 -0700 (PDT) 8, 24 -- Date: Tue, 18 Jul 2006 09:34:44 -0700 (PDT) 8, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 8, 24 -- To: microscopy-at-microscopy.com 8, 24 -- Subject: use of microwaves for sample preparation 8, 24 -- Message-ID: {Pine.SOC.4.64.0607180904180.9949-at-nature.Berkeley.EDU} 8, 24 -- MIME-Version: 1.0 8, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 8, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 ==============================End of - Headers==============================
The following position is available at Virginia Commonwealth University School of Medicine.
Microscopy Technician (Position # 55122)
A position is available in the Microscopy Facility of the Department of Anatomy and Neurobiology in the School of Medicine at Virginia Commonwealth University. The facility houses confocal, multi-photon, fluorescence, and electron microscopes (TEM & SEM). The successful candidate will assist with microscopy studies of various biological systems. Duties include instructing and assisting users of the facility, sample preparation, image analysis and routine equipment maintenance.
Applicants should have excellent communication and organizational skills, an understanding of basic laboratory procedures, and the ability to manage a large and varied workload. Qualifications include a degree in Biology/Life Sciences, a minimum of 2 years experience with laser scanning microscopy (confocal or multi-photon), sample preparation, digital imaging, and image analysis. Experience with electron microscopy is an asset. Computer skills are essential.
Applications are to be submitted online via the VCU Jobs website at: www.vcujobs.com
Click on the "Search Postings" link and under the "Working Title" field, select "Microscopy Technician".
--------------------------
Scott Henderson, Ph.D. Director of Microscopy Associate Professor Dept. Anatomy & Neurobiology Virginia Commonwealth University School of Medicine Sanger Hall, Rm. 9-069d 1101 East Marshall St. Richmond, VA 23298-0709
==============================Original Headers============================== 9, 28 -- From schenderson-at-vcu.edu Tue Jul 18 15:51:30 2006 9, 28 -- Received: from cedar.vcu.edu (cedar.vcu.edu [128.172.8.172]) 9, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6IKpUJv005000 9, 28 -- for {Microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 15:51:30 -0500 9, 28 -- Received: from Henderson ([128.172.29.122]) 9, 28 -- by cedar.vcu.edu (Lotus Domino Release 7.0.1FP1) 9, 28 -- with ESMTP id 2006071816512448-1271643 ; 9, 28 -- Tue, 18 Jul 2006 16:51:24 -0400 9, 28 -- Reply-To: {schenderson-at-vcu.edu} 9, 28 -- From: "Scott Henderson" {schenderson-at-vcu.edu} 9, 28 -- To: {Microscopy-at-microscopy.com} 9, 28 -- Subject: microscopy position available 9, 28 -- Date: Tue, 18 Jul 2006 16:51:23 -0400 9, 28 -- Organization: Virginia Commonwealth Unversity 9, 28 -- Message-ID: {002401c6aaab$f233d310$d361850a-at-Henderson} 9, 28 -- MIME-Version: 1.0 9, 28 -- X-Mailer: Microsoft Office Outlook 11 9, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 9, 28 -- Thread-Index: Acaqq+22V4y7UFr4SdO7xNFlc5sNMA== 9, 28 -- X-MIMETrack: Itemize by SMTP Server on CEDAR/VCU(Release 7.0.1FP1|April 17, 2006) at 07/18/2006 9, 28 -- 04:51:24 PM, 9, 28 -- Serialize by Router on CEDAR/VCU(Release 7.0.1FP1|April 17, 2006) at 07/18/2006 9, 28 -- 04:51:30 PM, 9, 28 -- Serialize complete at 07/18/2006 04:51:30 PM 9, 28 -- Content-Type: text/plain; 9, 28 -- charset="iso-8859-1" 9, 28 -- Content-Transfer-Encoding: 8bit 9, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6IKpUJv005000 ==============================End of - Headers==============================
I have a customer who would like to image in an SEM the crystal structure of PEEK (proprietary fluorocarbon polymer).
They can do it by light microscopy but the extra depth of focus in the SEM would be useful plus the ability to potentially ID inclusions by EDS.
Any suggestions? -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
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==============================Original Headers============================== 4, 14 -- From larry-at-cymru.freewire.co.uk Tue Jul 18 16:07:18 2006 4, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 4, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6IL7I08015287 4, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 18 Jul 2006 16:07:18 -0500 4, 14 -- Received: from [217.154.253.6] (th6dc-217-154-253-6.dial.mistral.co.uk [217.154.253.6]) 4, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k6IL7FDY021168 4, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 18 Jul 2006 22:07:16 +0100 4, 14 -- Mime-Version: 1.0 4, 14 -- Message-Id: {p06210200c0e2fcf1179a-at-[217.154.254.49]} 4, 14 -- Date: Tue, 18 Jul 2006 22:06:03 +0100 4, 14 -- To: Microscopy-at-MSA.Microscopy.Com 4, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 4, 14 -- Subject: Polymer Crystals by SEM? 4, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Does anyone know of an alternative to the Haskris R050 air cooled chiller for the LEO/Zeiss EMs? The Haskris unit I have is failing at least every year. The pump/motor unit will seize. Then the compressor control circuitry will fry.
Zeiss put special criteria on the chiller--45PSI, 30GPH, 72F. I don't see any other maker that can match this.
If they are there, please advise.
gary g.
==============================Original Headers============================== 5, 17 -- From gary-at-gaugler.com Tue Jul 18 20:14:25 2006 5, 17 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k6J1EOxO029041 5, 17 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 20:14:25 -0500 5, 17 -- Received: (qmail 30280 invoked from network); 18 Jul 2006 18:14:24 -0700 5, 17 -- Received: by simscan 1.1.0 ppid: 30277, pid: 30278, t: 0.0991s 5, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 5, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 5, 17 -- by qsmtp3 with SMTP; 18 Jul 2006 18:14:24 -0700 5, 17 -- Message-Id: {7.0.1.0.2.20060718181319.02462e60-at-gaugler.com} 5, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 5, 17 -- Date: Tue, 18 Jul 2006 18:14:25 -0700 5, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 5, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 5, 17 -- Subject: 5, 17 -- Mime-Version: 1.0 5, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Questions: Does anyone know of any companies that sell anti-BrdU antibodies that bind without the need for DNA denaturation? If not, is there an acid-free method for labelling cells in S-phase? And does anyone have a detergent-free method for labelling cells in M and S phases?
The details of this problem are as follows.
Details: I'm trying to determine is a protein toxin (directly labelled using a Molecular Probes Oregon Green 488 labelling kit) is internalised by a eukaryotic cell with respect to cell cycle. For S phase labelling I use BrdU incorporation plus a mouse monoclonal anti-BrdU from Zymed/Invitrogen. For M phase, I use murine anti-Phospho Histone H3, Ser10 (anti-PHH3). While labelling for cells in M/S phases has been successful, I have encountered a major problem with each labelling and I think they *may* be related.
Protocol: In short, non-synchronous populations of live cells are incubated with Oregon Green 488-labelled Stx (Stx-OG488) for 30min (and BrdU if labelling for cells in S phase), formalin fixed and permeabilised with 0.1% Triton X-100. For BrdU antigen retrieval, DNA is denatured for 30mins with 2M HCl and neutralised with Borax buffer. There after, M and S phase labellings are identical, in which the cells are blocked with PBS containing 20% FCS and 0.1% Triton. Primary antibodies (anti-BrdU and anti-PHH3) and secondaries conjugated to Alexa Fluor 594 are diluted in the blocking buffer and incubated for 1hr.
Hence, using dual fluorescence (green/red), we should be able to determine if cells in either M or S phase have a propensity to internalise Stx-OG488.
Here's the problem: Although visualising the cell phase is fine in the red field (texas red filter), I've found that the Stx-OG488 signal is a rather dull yellow or altogether missing when I switch to the green field (FITC filter).
Possible causes: We think this quenching is due to the use of acid denaturation for BrdU detection. I've tried 0.4M NaOH but this is highly alkaline and, as expected, also caused quenching of Stx-OG488. Also, treatment with DNase I -at- 10 U/ml for 1hr hasn't provided decent BrdU staining.
Furthermore, the continual presence of Triton X-100 during immunodetection for both M and S phases may cause dissociation of the toxin from the cell. I've tried lowering the Triton concentration and tried other detergents such as tween-20 and saponin with no improvement of the Stx-OG488 signal.
My questions are as follows: Are there any companies that sell anti-BrdU antibodies that bind without the need for DNA denaturation? If not, is there an acid-free method for labelling cells in S-phase? And does anyone have a detergent-free method for labelling cells in M and S phases?
I would be greatful for any suggestions/advice!
Thanks in advance, Damien Chong
Molecular Life Sciences Building Gate 8, Victoria Drive The University of Adelaide South Australia 5005
==============================Original Headers============================== 14, 29 -- From damien.chong-at-adelaide.edu.au Tue Jul 18 23:33:22 2006 14, 29 -- Received: from tosh.services.adelaide.edu.au (mailguard-send.adelaide.edu.au [192.43.227.21]) 14, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6J4XLsW009570 14, 29 -- for {microscopy-at-microscopy.com} ; Tue, 18 Jul 2006 23:33:22 -0500 14, 29 -- Received: from stimpy.services.adelaide.edu.au ([10.0.10.10]) 14, 29 -- by tosh.services.adelaide.edu.au with ESMTP; 19 Jul 2006 14:03:19 +0930 14, 29 -- X-IronPort-Anti-Spam-Filtered: true 14, 29 -- X-IronPort-Anti-Spam-Result: AQAAAAdTvUQN 14, 29 -- X-IronPort-AV: i="4.06,257,1149431400"; 14, 29 -- d="scan'208"; a="7422374:sNHT71535387" 14, 29 -- Received: from stimpy.services.adelaide.edu.au (localhost.localdomain [127.0.0.1]) 14, 29 -- by stimpy.services.adelaide.edu.au (8.13.1/8.13.1) with ESMTP id k6J4XJpE012077 14, 29 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 14:03:19 +0930 14, 29 -- Received: (from apache-at-localhost) 14, 29 -- by stimpy.services.adelaide.edu.au (8.13.1/8.13.1/Submit) id k6J4XJcT012068 14, 29 -- for microscopy-at-microscopy.com; Wed, 19 Jul 2006 14:03:19 +0930 14, 29 -- Received: from molb00325.molecularbiosciences.adelaide.edu.au (molb00325.molecularbiosciences.adelaide.edu.au [129.127.106.33]) 14, 29 -- by webmail.adelaide.edu.au (IMP) with HTTP 14, 29 -- for {dchong01-at-localhost} ; Wed, 19 Jul 2006 14:03:19 +0930 14, 29 -- Message-ID: {1153283599.44bdb60f20c60-at-webmail.adelaide.edu.au} 14, 29 -- Date: Wed, 19 Jul 2006 14:03:19 +0930 14, 29 -- From: Damien Chong {damien.chong-at-adelaide.edu.au} 14, 29 -- To: Microscopy ListServer {microscopy-at-microscopy.com} 14, 29 -- Subject: LM (IF): Quenching(?) with mitosis and S phase labelling 14, 29 -- MIME-Version: 1.0 14, 29 -- Content-Type: text/plain; charset=ISO-8859-1 14, 29 -- Content-Transfer-Encoding: 8bit 14, 29 -- User-Agent: Internet Messaging Program (IMP) 3.2.6 14, 29 -- X-Originating-IP: 129.127.106.33 ==============================End of - Headers==============================
I think that the antibody cannot access native DNA (and so incorporated BrU) because it is covered by proteins, so your problem does not come from the antibody (Anti-BrdU antibody are very good, at least the one from Boehringer I tried was excellent). You have to denature the DNA in order to give access to the DNA. For the permeabilization and detergent problem, you can try methanol fixation at -20°C, which permeabilizes the cells at the same time. For the detection, don't use Triton, but use serum or gelatin or fat-free milk powder to minimize non-specific interactions. If your secondary antibodies are from goat, use goat serum, not bovine serum!
Good luck,
Stephane
--- damien.chong-at-adelaide.edu.au wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Send To: Microscopy-at-Microscopy.Com } } Hi Listers, } } Questions: Does anyone know of any companies that } sell anti-BrdU antibodies } that bind without the need for DNA denaturation? If } not, is there an acid-free } method for labelling cells in S-phase? And does } anyone have a detergent-free } method for labelling cells in M and S phases? } } The details of this problem are as follows. } } Details: } I'm trying to determine is a protein toxin (directly } labelled using a Molecular } Probes Oregon Green 488 labelling kit) is } internalised by a eukaryotic cell } with respect to cell cycle. For S phase labelling I } use BrdU incorporation } plus a mouse monoclonal anti-BrdU from } Zymed/Invitrogen. For M phase, I use } murine anti-Phospho Histone H3, Ser10 (anti-PHH3). } While } labelling for cells in M/S phases has been } successful, I have encountered a } major problem with each labelling and I think they } *may* be related. } } Protocol: } In short, non-synchronous populations of live cells } are incubated with Oregon } Green 488-labelled Stx (Stx-OG488) for 30min (and } BrdU if labelling for cells } in S phase), formalin fixed and permeabilised with } 0.1% Triton X-100. For BrdU } antigen retrieval, DNA is denatured for 30mins with } 2M HCl and neutralised with } Borax buffer. There after, M and S phase labellings } are identical, in which } the cells are blocked with PBS containing 20% FCS } and 0.1% Triton. Primary } antibodies (anti-BrdU and anti-PHH3) and secondaries } conjugated to Alexa Fluor } 594 are diluted in the blocking buffer and incubated } for 1hr. } } Hence, using dual fluorescence (green/red), we } should be able to determine if } cells in either M or S phase have a propensity to } internalise Stx-OG488. } } Here's the problem: } Although visualising the cell phase is fine in the } red field (texas red } filter), I've found that the Stx-OG488 signal is a } rather dull yellow or } altogether missing when I switch to the green field } (FITC filter). } } Possible causes: } We think this quenching is due to the use of acid } denaturation for BrdU } detection. I've tried 0.4M NaOH but this is highly } alkaline and, as expected, } also caused quenching of Stx-OG488. Also, treatment } with DNase I -at- 10 U/ml for } 1hr hasn't provided decent BrdU staining. } } Furthermore, the continual presence of Triton X-100 } during immunodetection for } both M and S phases may cause dissociation of the } toxin from the cell. I've } tried lowering the Triton concentration and tried } other detergents such as } tween-20 and saponin with no improvement of the } Stx-OG488 signal. } } My questions are as follows: } Are there any companies that sell anti-BrdU } antibodies that bind without the } need for DNA denaturation? If not, is there an } acid-free method for labelling } cells in S-phase? And does anyone have a } detergent-free method for labelling } cells in M and S phases? } } I would be greatful for any suggestions/advice! } } Thanks in advance, } Damien Chong } } Molecular Life Sciences Building } Gate 8, Victoria Drive } The University of Adelaide } South Australia 5005 } } ==============================Original } Headers============================== } 14, 29 -- From damien.chong-at-adelaide.edu.au Tue Jul } 18 23:33:22 2006 } 14, 29 -- Received: from } tosh.services.adelaide.edu.au } (mailguard-send.adelaide.edu.au [192.43.227.21]) } 14, 29 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k6J4XLsW009570 } 14, 29 -- for {microscopy-at-microscopy.com} ; Tue, 18 } Jul 2006 23:33:22 -0500 } 14, 29 -- Received: from } stimpy.services.adelaide.edu.au ([10.0.10.10]) } 14, 29 -- by tosh.services.adelaide.edu.au with } ESMTP; 19 Jul 2006 14:03:19 +0930 } 14, 29 -- X-IronPort-Anti-Spam-Filtered: true } 14, 29 -- X-IronPort-Anti-Spam-Result: AQAAAAdTvUQN } 14, 29 -- X-IronPort-AV: i="4.06,257,1149431400"; } 14, 29 -- d="scan'208"; a="7422374:sNHT71535387" } 14, 29 -- Received: from } stimpy.services.adelaide.edu.au } (localhost.localdomain [127.0.0.1]) } 14, 29 -- by stimpy.services.adelaide.edu.au } (8.13.1/8.13.1) with ESMTP id k6J4XJpE012077 } 14, 29 -- for {microscopy-at-microscopy.com} ; Wed, 19 } Jul 2006 14:03:19 +0930 } 14, 29 -- Received: (from apache-at-localhost) } 14, 29 -- by stimpy.services.adelaide.edu.au } (8.13.1/8.13.1/Submit) id k6J4XJcT012068 } 14, 29 -- for microscopy-at-microscopy.com; Wed, 19 } Jul 2006 14:03:19 +0930 } 14, 29 -- Received: from } molb00325.molecularbiosciences.adelaide.edu.au } (molb00325.molecularbiosciences.adelaide.edu.au } [129.127.106.33]) } 14, 29 -- by webmail.adelaide.edu.au (IMP) with } HTTP } 14, 29 -- for {dchong01-at-localhost} ; Wed, 19 Jul } 2006 14:03:19 +0930 } 14, 29 -- Message-ID: } {1153283599.44bdb60f20c60-at-webmail.adelaide.edu.au} } 14, 29 -- Date: Wed, 19 Jul 2006 14:03:19 +0930 } 14, 29 -- From: Damien Chong } {damien.chong-at-adelaide.edu.au} } 14, 29 -- To: Microscopy ListServer } {microscopy-at-microscopy.com} } 14, 29 -- Subject: LM (IF): Quenching(?) with } mitosis and S phase labelling } 14, 29 -- MIME-Version: 1.0 } 14, 29 -- Content-Type: text/plain; } charset=ISO-8859-1 } 14, 29 -- Content-Transfer-Encoding: 8bit } 14, 29 -- User-Agent: Internet Messaging Program } (IMP) 3.2.6 } 14, 29 -- X-Originating-IP: 129.127.106.33 } ==============================End of - } Headers============================== }
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This Question was submitted to Ask-A-Microscopist by (lgauthier-at-victrex.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, July 19, 2006 at 04:07:13 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both lgauthier-at-victrex.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: Dear Sir or Madam, I would like to know if it is possible to observe with a SEM the morphology (spherolites, cristals...) of a polymer (for my current study it is a PEEK). If yes in which conditions (HV or LV, pressure etc..) I should work? Thank you very much in advance.
Hello everybody, Recently I received samples that I processed for SEM (oysters). When I finished the last step of processing sputter coating I saw that my samples where not coated but the stubs were. I had a good vacuum (around 10-1 mbar), tank full of argon, current 10 mA, and I coated 6 min total. Would any body have any idea what might be the problem? It's needless to say that scoping is impossible due to charging. Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Wed Jul 19 09:24:18 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JEOHBT016664 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 09:24:18 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1G3CyE-0004oK-01 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 11:24:14 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 19 Jul 06 11:24:14 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 19 Jul 06 11:23:02 -0300 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Wed, 19 Jul 2006 11:20:29 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: Sputter coating problems 1, 22 -- Message-ID: {44BE157D.12915.774EA9-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
I expect that you will be disappointed. Typically, elucidation of the morphology of crystalline morphology of polymers generally requires etching or staining.
Regards,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
lgauthier-at-vict rex.com To gary.m.brown-at-exxonmobil.com 07/19/06 06:39 cc AM Subject [Microscopy] AskAMicroscopist: Please respond Observation of polymers morphology to lgauthier-at-vict rex.com
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This Question was submitted to Ask-A-Microscopist by (lgauthier-at-victrex.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, July 19, 2006 at 04:07:13 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both lgauthier-at-victrex.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: Dear Sir or Madam, I would like to know if it is possible to observe with a SEM the morphology (spherolites, cristals...) of a polymer (for my current study it is a PEEK). If yes in which conditions (HV or LV, pressure etc..) I should work? Thank you very much in advance.
We have an old Lab-line/Hooker Plant Microtome, cat #1225 with a fried motor. Lab-line is no more, and the current company (Barnstead, I think) no longer has parts for these. Does anyone have any spares or a motor or know of a source. We've been checking the used lab equipment people to no avail. Thanks.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Wed Jul 19 10:15:53 2006 3, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JFFqwF005507 3, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 10:15:52 -0500 3, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k6JFnNfd021702 3, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 11:49:28 -0400 3, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 20 -- Wed, 19 Jul 2006 11:15:50 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f0623090ac0e3fca33212-at-[141.209.160.132]} 3, 20 -- Date: Wed, 19 Jul 2006 11:15:50 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: Hooker plant microtome parts 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 19 Jul 2006 15:15:50.0637 (UTC) FILETIME=[383F49D0:01C6AB46] 3, 20 -- X-CanItPRO-Stream: default 3, 20 -- X-Spam-Score: -3.2 () L_EXCH_MF,PORN_RP_HOOKERS_ 3, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
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I have had this occur (with various specimen materials). I have not investigated it, but my first guess is significant off-gassing of the sample. Depending on the gauging configuration/location and vacuum pump capacity, it is entirely possible to indicate a good vacuum, but this may not be true in the local area of the specimen. Try pumping for quite a while* (best to slightly warm the specimen) to outgas - then try to coat.
*- I have an untrapped rough pump (only) coater, so I leak a low partial pressure of argon while long term pumping to minimize pump oil contamination.
Woody White BWXT Services
-----Original Message----- X-from: wadowska-at-upei.ca [mailto:wadowska-at-upei.ca] Sent: Wednesday, July 19, 2006 10:25 AM To: White, Woody N.
Hello everybody, Recently I received samples that I processed for SEM (oysters). When I finished the last step of processing sputter coating I saw that my samples where not coated but the stubs were. I had a good vacuum (around 10-1 mbar), tank full of argon, current 10 mA, and I coated 6 min total. Would any body have any idea what might be the problem? It's needless to say that scoping is impossible due to charging. Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Wed Jul 19 09:24:18 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JEOHBT016664 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 09:24:18 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1G3CyE-0004oK-01 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 11:24:14 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 19 Jul 06 11:24:14 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 19 Jul 06 11:23:02 -0300 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-microscopy.com 1, 22 -- Date: Wed, 19 Jul 2006 11:20:29 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: Sputter coating problems 1, 22 -- Message-ID: {44BE157D.12915.774EA9-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
==============================Original Headers============================== 13, 26 -- From nwwhite-at-bwxt.com Wed Jul 19 13:00:39 2006 13, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 13, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JI0cdx028557 13, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 13:00:39 -0500 13, 26 -- Received: from ([131.184.13.224]) 13, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.1733907; 13, 26 -- Wed, 19 Jul 2006 14:00:29 -0400 13, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 13, 26 -- Wed, 19 Jul 2006 14:00:28 -0400 13, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 26 -- Content-class: urn:content-classes:message 13, 26 -- MIME-Version: 1.0 13, 26 -- Content-Type: text/plain; 13, 26 -- charset="US-ASCII" 13, 26 -- Subject: RE: [Microscopy] Sputter coating problems 13, 26 -- Date: Wed, 19 Jul 2006 14:00:28 -0400 13, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703CFFC-at-BWXSPO01.BWXS.BWXTECH.NET} 13, 26 -- X-MS-Has-Attach: 13, 26 -- X-MS-TNEF-Correlator: 13, 26 -- Thread-Topic: [Microscopy] Sputter coating problems 13, 26 -- thread-index: AcarPzmGxRaSPq8CQJGntw3be2JC/wAHMyQg 13, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 13, 26 -- To: {wadowska-at-upei.ca} , "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 13, 26 -- X-OriginalArrivalTime: 19 Jul 2006 18:00:28.0839 (UTC) FILETIME=[381D4B70:01C6AB5D] 13, 26 -- Content-Transfer-Encoding: 8bit 13, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6JI0cdx028557 ==============================End of - Headers==============================
Sorry I misspelled your name before. A senior moment, I suppose.
There is a variety of tools that may be used to study the crystalline morphology of polymers. Confocal microscopy, as suggested by Tony, may be use. The choice of tool will depend on several factors including the form of the polymer (reactor particles, molded parts, blown/cast films, etc.), the scale of the structures to be analyzed (spherulites, crystallites/lamellae), the tools available for sample preparation, and the type of SEM you use (conventional, variable pressure, or field emission SEM).
Suggest you check out the latest edition of Sawyer and Grubbs Polymer Morphology. This book is a great place to start and should have a place on every lab's book shelf.
Best of luck,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com ----- Forwarded by Gary M Brown/Baytown/ExxonMobil on 07/19/06 01:31 PM -----
gary.m.brown-at-e xxonmobil.com To gary.m.brown-at-exxonmobil.com 07/19/06 10:09 cc AM Subject [Microscopy] Re: AskAMicroscopist: Please respond Observation of polymers morphology to gary.m.brown-at-e xxonmobil.com
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Igauthier,
I expect that you will be disappointed. Typically, elucidation of the morphology of crystalline morphology of polymers generally requires etching or staining.
Regards,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
lgauthier-at-vict rex.com To gary.m.brown-at-exxonmobil.com 07/19/06 06:39 cc AM Subject [Microscopy] AskAMicroscopist: Please respond Observation of polymers morphology to lgauthier-at-vict rex.com
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
This Question was submitted to Ask-A-Microscopist by (lgauthier-at-victrex.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, July 19, 2006 at 04:07:13 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both lgauthier-at-victrex.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: Dear Sir or Madam, I would like to know if it is possible to observe with a SEM the morphology (spherolites, cristals...) of a polymer (for my current study it is a PEEK). If yes in which conditions (HV or LV, pressure etc..) I should work? Thank you very much in advance.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both smythen-at-musc.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: smythen-at-musc.edu Name: Nancy Smythe
Organization: Medical University of South Carolina
Title-Subject: [Filtered] help finding a book
Question: I am looking for a book that is out of print. The name is "Ultrastructural Atlas of the Inner Ear" Edited by Friedmann and Ballantyne. I had a copy but like things in labs do it grew legs and walked! Thanks
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both shaenon-at-hotmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] LKB 8800A-NM Ultrotome III Problem
Question: Hi, I am having a problem with my ultramicrotome. The specimen arm appears to be stuck in the "down" position. I cannot get it to return to the original position. I can gently push the arm up with my hand but it immediately falls back down when I let go. Does anyone have any suggestions on how I may mix this?
PS - I have tried switched back and forth between manual and automatic modes, locked the arm and returning it to the "free" position.
Thanks for the on- and off-list replies. Very helpful as usual.
An additive is definitely needed. Haskris says no--just use distilled water. that does not work for long. I use 10% mix of Ethylene Glycol (1/2G to 4.5G distilled water). That works fine. I change the nylon and toilet paper filters about once a year and change out the water mix. It is also good practice to periodically check the nylon filter for signs of algae.
The R050 chillers for Zeiss/LEO are built with quite a few options. One of these is option G. This is a hot gas bypass system. The net effect of this is that the compressor runs continuously at the factory set temperature (65F). The temperature can be changed but the compressor will still run continuously. When Zeiss installed the system, they essentially bypassed this option. They changed the thermostat set point to 72F. Doing so causes the compressor to cycle. However, by using the turbine pump which is always running, compressor on/off is not seen at the SEM. The rationale for doing this change was to bring the chilled water closer to ambient temperature of the SEM to avoid condensation.
Zeiss uses a cooling design for its electronics that IMO is much more sophisticated than others. The consequence of this is that the chiller needs to have a larger reservoir (5G) and higher flow rate. but with pumps and chiller in separate rooms from the SEM, the heat output from the SEM is very low.
All feedback about Haskris has been hugely positive. My experiences with Haskris people is also very positive. So I will stick with them.
My problem turns out to be a blown compressor rather than a seized pump. The pump motor will not operate since the high temp sensor disables the motor. When the water cools down a little from the limit, the motor runs OK. The compressor is the problem. The windings are open and the current relay is basically vaporized. So the whole compressor has to be replaced. Has anyone seen this problem before? The only known rationale for this is low line voltage. In my case, the chiller is on a Liebert Nfinity 8KVA UPS. Since I always run in inverter mode, the chiller will always get perfect power. The UPS will take out any line sags or spikes.
The concept of a single point of failure is in operation....sigh.
gary g.
At 08:30 AM 7/19/2006, you wrote:
} Gary, I have a LEO/Zeiss SEM and also the same chiller. My previous } experience } with Haskriss chillers is that some sort of water additive must be } used to eliminate } bacteria growth and corrosion. When our 1550VP was first installed } we had problems } with clogging filters and thermal alarms on the SEM until we started } treating the water. } I have been using the same additive that our building chilled water } system engineers } use at about 4 ounces per tank. I have absolutely no idea what is in } it, but we no longer } have filter issues and the Haskriss runs continuously without any } sign of corrosion } anywhere. } } } } } } Does anyone know of an alternative to the Haskris R050 air cooled } chiller for the LEO/Zeiss EMs? The Haskris unit I have is failing at } least every year. The pump/motor unit will seize. Then the } compressor control circuitry will fry. } } Zeiss put special criteria on the chiller--45PSI, 30GPH, 72F. I } don't see any other maker that can match this. } } If they are there, please advise. } } gary g.
1. The cord or wire that raises and lowers the arm may be broken, slipped or stretched. Best way to determine this is to remove the front plate covering over the arm and see if the cord is intact. When you turn the manual knob, does anything move? Look specifically for the motor where the cord ends. You should see the motor turn as you move the knob. If the motor does not move, go to step 2.
2. The toothed belt that goes from the manual control knob to a small potentiometer (that raises and lowers the arm) may be worn out. Over time, this belt rots and just disintegrates. You may be moving the large manual knob but the movement is not being transferred to the potentiometer. Best way to determine this is to remove the top (3-sided, dark blue) cover from the control unit. Look for the belt that connects to the potentiometer from the manual knob. If broken, it needs to be replaced. Good luck finding one as I think they are no longer available. However, you may be able to get a similar one from a machine supply house. Check back with this listserver, as well, since someone may have one to spare. Be careful when removing covers to AVOID GETTING ELECTRICAL SHOCKS. Observe but don't probe around amongst the electronics.
3. Lastly, the potentiometer itself may be defective (if the belt is in place and moving the potentiometer control and nothing is happening with specimen arm). Another possibility: sometimes the potentiometers seize up and the belt gets stripped when it tries to turn it. Do you have any undue resistance when you move the manual control on the front of the control unit?
Try these diagnostics, take an aspirin and call me back in the morning if you have persistent problems. We are glad to help.
JB
} } Title-Subject: [Filtered] LKB 8800A-NM Ultrotome III Problem } } Question: Hi, I am having a problem with my ultramicrotome. The } specimen arm appears to be stuck in the "down" position. I cannot } get it to return to the original position. I can gently push the arm } up with my hand but it immediately falls back down when I let go. } Does anyone have any suggestions on how I may mix this? } } PS - I have tried switched back and forth between manual and } automatic modes, locked the arm and returning it to the "free" } position. } } Thanks, } Shannon } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 8, 12 -- From zaluzec-at-microscopy.com Wed Jul 19 14:44:43 2006 } 8, 12 -- Received: from [10.255.254.48] (msdvpn26.msd.anl.gov } [130.202.238.90]) } 8, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k6JJiffv020179 } 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 } 14:44:42 -0500 } 8, 12 -- Mime-Version: 1.0 } 8, 12 -- X-Sender: (Unverified) } 8, 12 -- Message-Id: {p06020406c0e43c1982c3-at-[10.255.254.48]} } 8, 12 -- Date: Wed, 19 Jul 2006 15:44:43 -0400 } 8, 12 -- To: microscopy-at-microscopy.com } 8, 12 -- From: shaenon-at-hotmail.com (by way of MicroscopyListserver) } 8, 12 -- Subject: viaWWW: LKB 8800A-NM Ultrotome III Problem } 8, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers==============================
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 12, 18 -- From bozzola-at-siu.edu Wed Jul 19 15:32:20 2006 12, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 12, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JKWKwA016808 12, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 15:32:20 -0500 12, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 12, 18 -- by abbmta2.siu.edu (Switch-3.1.9/Switch-3.1.7) with ESMTP id k6JKWGsJ019609 12, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 15:32:18 -0500 (CDT) 12, 18 -- Mime-Version: 1.0 12, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 12, 18 -- Message-Id: {p0611040bc0e4433498fd-at-[131.230.177.142]} 12, 18 -- In-Reply-To: {200607191945.k6JJjSfq022550-at-ns.microscopy.com} 12, 18 -- References: {200607191945.k6JJjSfq022550-at-ns.microscopy.com} 12, 18 -- Date: Wed, 19 Jul 2006 15:32:15 -0500 12, 18 -- To: Microscopy-at-msa.microscopy.com 12, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 12, 18 -- Subject: Re: [Microscopy] viaWWW: LKB 8800A-NM Ultrotome III Problem 12, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 12, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
We have an LKB Knifemaker Model 7801A that just stopped making knives. I had recently adjusted it, replaced the cutting wheel, lubricated a couple parts, etc., and it was making knives like gangbusters. About 3 weeks later the locking arm (the lever on the left that locks the scoring apparatus down) seemed to become looser and easily depresses to a lower point than it ever did before. It doesn't seem to be applying enough pressure on the cutting arm. It's still scoring, but turning the breaking wheel (the one on lower right that applies pressure to the scored glass) doesn't cause the glass to break.
Does anyone have a clue about how to fix this, or even what went wrong, from my probably garbled description? Are there any schematics out there that someone might be willing to share? Is there any relief from this heat?
Again, the only thing that seems to have changed is the locking arm feeling looser and it's not obvious to me how to adjust this.
Thanks for any advice you might have! I didn't think you could damage one of these things with an Abrams tank....
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well When Our Knifebreaker Works! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Wed Jul 19 16:01:36 2006 10, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JL1ZCP027508 10, 23 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 16:01:35 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Wed, 19 Jul 2006 16:01:30 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: LKB Knifemaker on the skids 10, 23 -- Date: Wed, 19 Jul 2006 16:01:31 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E338E-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: LKB Knifemaker on the skids 10, 23 -- thread-index: AcardoOcvNcWH+D4QRmtpEpa94Gopg== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 19 Jul 2006 21:01:30.0992 (UTC) FILETIME=[82764F00:01C6AB76] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6JL1ZCP027508 ==============================End of - Headers==============================
Many years ago, we developed a technique for etching PEEK for observation under TEM. But it would also work under a good modern SEM. Here are the two papers. If you want any more details, please contact me at R.H.Olley-at-reading.ac.uk .
On Crystallization Phenomena in PEEK Bassett,D.C., Olley,R.H., Al Raheil,I.A.M. Polymer, 1988, vol.29, pp.1745-1754
----------------------------------- Robert H. Olley Reply to: R.H.Olley-at-reading.ac.uk URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
==============================Original Headers============================== 6, 23 -- From hinmeigeng-at-hotmail.com Wed Jul 19 16:01:45 2006 6, 23 -- Received: from bay0-omc2-s33.bay0.hotmail.com (bay0-omc2-s33.bay0.hotmail.com [65.54.246.169]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JL1j4E027635 6, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 19 Jul 2006 16:01:45 -0500 6, 23 -- Received: from hotmail.com ([64.4.56.23]) by bay0-omc2-s33.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Wed, 19 Jul 2006 14:01:44 -0700 6, 23 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 6, 23 -- Wed, 19 Jul 2006 14:01:43 -0700 6, 23 -- Message-ID: {BAY101-F1328BD6B77428118F7CCFFCA600-at-phx.gbl} 6, 23 -- Received: from 64.4.56.200 by by101fd.bay101.hotmail.msn.com with HTTP; 6, 23 -- Wed, 19 Jul 2006 21:01:41 GMT 6, 23 -- X-Originating-IP: [86.128.172.175] 6, 23 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 6, 23 -- X-Sender: hinmeigeng-at-hotmail.com 6, 23 -- Reply-To: R.H.Olley-at-reading.ac.uk 6, 23 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 6, 23 -- To: Microscopy-at-MSA.Microscopy.Com 6, 23 -- Cc: R.H.Olley-at-reading.ac.uk 6, 23 -- Subject: Re: AskAMicroscopist: Observation of polymers morphology 6, 23 -- Date: Wed, 19 Jul 2006 21:01:41 +0000 6, 23 -- Mime-Version: 1.0 6, 23 -- Content-Type: text/plain; format=flowed 6, 23 -- X-OriginalArrivalTime: 19 Jul 2006 21:01:43.0828 (UTC) FILETIME=[8A1CED40:01C6AB76] ==============================End of - Headers==============================
What is happening is that the locking head is not locking it tight enough. This is caused by people (not you, of course, as I know that you know better) consistently overtightening the locking lever. If the lever is pushed all the way down so that the black ball is touching the base plate, it can no longer exert enough pressure to clamp the head in place. I believe that you can insert a metal washer on (the end of?) the shaft of the tightening rod and this will allow more pressure to be applied. But the big NO, NO here is (after it is fixed) not to push the arm all the way down to lock it in place. You stop pushing down when you feel resistance. Ideally, it should be safely locked with the black ball about 1 inch above the base plate.
You going to M&M?
JB
} We have an LKB Knifemaker Model 7801A that just stopped making knives. } I had recently adjusted it, replaced the cutting wheel, lubricated a } couple parts, etc., and it was making knives like gangbusters. About 3 } weeks later the locking arm (the lever on the left that locks the } scoring apparatus down) seemed to become looser and easily depresses to } a lower point than it ever did before. It doesn't seem to be applying } enough pressure on the cutting arm. It's still scoring, but turning the } breaking wheel (the one on lower right that applies pressure to the } scored glass) doesn't cause the glass to break. } } Does anyone have a clue about how to fix this, or even what went wrong, } from my probably garbled description? Are there any schematics out } there that someone might be willing to share? Is there any relief from } this heat? } } Again, the only thing that seems to have changed is the locking arm } feeling looser and it's not obvious to me how to adjust this. } } Thanks for any advice you might have! I didn't think you could damage } one of these things with an Abrams tank.... } } Cheers, } Randy } } Randy Tindall } EM Specialist } Electron Microscopy Core Facility---We Do Small Well When Our } Knifebreaker Works! } W125 Veterinary Medicine } University of Missouri } Columbia, MO 65211 } Tel: (573) 882-8304 } Fax: (573) 884-2227 } Email: tindallr-at-missouri.edu } Web: http://www.emc.missouri.edu } } } } } } } } ==============================Original Headers============================== } 10, 23 -- From TindallR-at-missouri.edu Wed Jul 19 16:01:36 2006 } 10, 23 -- Received: from um-nsmtpout1.um.umsystem.edu } (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) } 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k6JL1ZCP027508 } 10, 23 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 } 16:01:35 -0500 } 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu } ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft } SMTPSVC(6.0.3790.1830); } 10, 23 -- Wed, 19 Jul 2006 16:01:30 -0500 } 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 10, 23 -- Content-class: urn:content-classes:message } 10, 23 -- MIME-Version: 1.0 } 10, 23 -- Content-Type: text/plain; } 10, 23 -- charset="us-ascii" } 10, 23 -- Subject: LKB Knifemaker on the skids } 10, 23 -- Date: Wed, 19 Jul 2006 16:01:31 -0500 } 10, 23 -- Message-ID: } {91108EF9255B394CBF8B7E3789814A411E338E-at-UM-XMAIL08.um.umsystem.edu} } 10, 23 -- X-MS-Has-Attach: } 10, 23 -- X-MS-TNEF-Correlator: } 10, 23 -- Thread-Topic: LKB Knifemaker on the skids } 10, 23 -- thread-index: AcardoOcvNcWH+D4QRmtpEpa94Gopg== } 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} } 10, 23 -- To: {microscopy-at-microscopy.com} } 10, 23 -- X-OriginalArrivalTime: 19 Jul 2006 21:01:30.0992 (UTC) } FILETIME=[82764F00:01C6AB76] } 10, 23 -- Content-Transfer-Encoding: 8bit } 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k6JL1ZCP027508 } ==============================End of - Headers==============================
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 8, 18 -- From bozzola-at-siu.edu Wed Jul 19 16:20:17 2006 8, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JLKHj6015591 8, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 16:20:17 -0500 8, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 8, 18 -- by abbmta2.siu.edu (Switch-3.1.9/Switch-3.1.7) with ESMTP id k6JLKGIF001208 8, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 16:20:16 -0500 (CDT) 8, 18 -- Mime-Version: 1.0 8, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 8, 18 -- Message-Id: {p0611040ec0e450cdc906-at-[131.230.177.142]} 8, 18 -- In-Reply-To: {200607192103.k6JL3JkD031100-at-ns.microscopy.com} 8, 18 -- References: {200607192103.k6JL3JkD031100-at-ns.microscopy.com} 8, 18 -- Date: Wed, 19 Jul 2006 16:20:15 -0500 8, 18 -- To: Microscopy-at-msa.microscopy.com 8, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 8, 18 -- Subject: Re: [Microscopy] LKB Knifemaker on the skids 8, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 8, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
One last comment. Haskris offers, in addition to air- and water-cooled chillers, a heat exchanger for lower volume cooling needs. It depends solely on heat exchange between a supply of cool water and the closed system cooling the microscope. One significant limitation is that it depends on a cool water supply. Here in Texas, our supply of surface water in the summer can easily approach nearly 90F; too high a temperature for this heat exchanger to do much cooling. I suggest that you check out this heat exchanger if you work in an area where the tap water is cool to cold and the heat from your equipment is modest.
Regarding the need for an algaecide in chillers: We use opaque black hose for the cooling lines of our microscopes. This eliminates any source of light that can support growth of algae. Never use tubing of any kind. The hose is also heavier than tubing and resists hydrodynamic failure.
Practicing this over the past 20 years, we have never encountered problems with algae. Fill the chiller with deionized water and you're good to go.
Regards,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
gary-at-gaugler.c om To gary.m.brown-at-exxonmobil.com 07/19/06 03:19 cc PM Subject [Microscopy] Re: Haskris chiller for Please respond Zeiss/LEO to gary-at-gaugler.c om
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Thanks for the on- and off-list replies. Very helpful as usual.
An additive is definitely needed. Haskris says no--just use distilled water. that does not work for long. I use 10% mix of Ethylene Glycol (1/2G to 4.5G distilled water). That works fine. I change the nylon and toilet paper filters about once a year and change out the water mix. It is also good practice to periodically check the nylon filter for signs of algae.
The R050 chillers for Zeiss/LEO are built with quite a few options. One of these is option G. This is a hot gas bypass system. The net effect of this is that the compressor runs continuously at the factory set temperature (65F). The temperature can be changed but the compressor will still run continuously. When Zeiss installed the system, they essentially bypassed this option. They changed the thermostat set point to 72F. Doing so causes the compressor to cycle. However, by using the turbine pump which is always running, compressor on/off is not seen at the SEM. The rationale for doing this change was to bring the chilled water closer to ambient temperature of the SEM to avoid condensation.
Zeiss uses a cooling design for its electronics that IMO is much more sophisticated than others. The consequence of this is that the chiller needs to have a larger reservoir (5G) and higher flow rate. but with pumps and chiller in separate rooms from the SEM, the heat output from the SEM is very low.
All feedback about Haskris has been hugely positive. My experiences with Haskris people is also very positive. So I will stick with them.
My problem turns out to be a blown compressor rather than a seized pump. The pump motor will not operate since the high temp sensor disables the motor. When the water cools down a little from the limit, the motor runs OK. The compressor is the problem. The windings are open and the current relay is basically vaporized. So the whole compressor has to be replaced. Has anyone seen this problem before? The only known rationale for this is low line voltage. In my case, the chiller is on a Liebert Nfinity 8KVA UPS. Since I always run in inverter mode, the chiller will always get perfect power. The UPS will take out any line sags or spikes.
The concept of a single point of failure is in operation....sigh.
gary g.
At 08:30 AM 7/19/2006, you wrote:
} Gary, I have a LEO/Zeiss SEM and also the same chiller. My previous } experience } with Haskriss chillers is that some sort of water additive must be } used to eliminate } bacteria growth and corrosion. When our 1550VP was first installed } we had problems } with clogging filters and thermal alarms on the SEM until we started } treating the water. } I have been using the same additive that our building chilled water } system engineers } use at about 4 ounces per tank. I have absolutely no idea what is in } it, but we no longer } have filter issues and the Haskriss runs continuously without any } sign of corrosion } anywhere. } } } } } } Does anyone know of an alternative to the Haskris R050 air cooled } chiller for the LEO/Zeiss EMs? The Haskris unit I have is failing at } least every year. The pump/motor unit will seize. Then the } compressor control circuitry will fry. } } Zeiss put special criteria on the chiller--45PSI, 30GPH, 72F. I } don't see any other maker that can match this. } } If they are there, please advise. } } gary g.
Randy, We have a model 7800 knife maker. If these models are similar, here's what I think might have happened. When the unit was lubricated, it has allowed the adjusting collar on the tightening handle shaft to move (round collar with circular holes in it for a spanner wrench). Simply turning this adjusting collar to the correct position will correct the problem. Maybe removing some lubricant too!
Good luck, Kevin
Kevin Battjes Impact Analytical Voice 989-832-5555, ext 556 Michigan Molecular Institute Fax 989-832-5560 1910 W. St Andrews Road e-mail: battjes-at-mmi.org Midland MI 48640 battjes-at-impactanalytical.com
-----Original Message----- X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] Sent: Wednesday, July 19, 2006 5:07 PM To: battjes-at-impactanalytical.com
We have an LKB Knifemaker Model 7801A that just stopped making knives. I had recently adjusted it, replaced the cutting wheel, lubricated a couple parts, etc., and it was making knives like gangbusters. About 3 weeks later the locking arm (the lever on the left that locks the scoring apparatus down) seemed to become looser and easily depresses to a lower point than it ever did before. It doesn't seem to be applying enough pressure on the cutting arm. It's still scoring, but turning the breaking wheel (the one on lower right that applies pressure to the scored glass) doesn't cause the glass to break.
Does anyone have a clue about how to fix this, or even what went wrong, from my probably garbled description? Are there any schematics out there that someone might be willing to share? Is there any relief from this heat?
Again, the only thing that seems to have changed is the locking arm feeling looser and it's not obvious to me how to adjust this.
Thanks for any advice you might have! I didn't think you could damage one of these things with an Abrams tank....
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well When Our Knifebreaker Works! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Wed Jul 19 16:01:36 2006 10, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JL1ZCP027508 10, 23 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 16:01:35 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Wed, 19 Jul 2006 16:01:30 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: LKB Knifemaker on the skids 10, 23 -- Date: Wed, 19 Jul 2006 16:01:31 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E338E-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: LKB Knifemaker on the skids 10, 23 -- thread-index: AcardoOcvNcWH+D4QRmtpEpa94Gopg== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 19 Jul 2006 21:01:30.0992 (UTC) FILETIME=[82764F00:01C6AB76] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6JL1ZCP027508 ==============================End of - Headers==============================
==============================Original Headers============================== 22, 16 -- From battjes-at-impactanalytical.com Wed Jul 19 16:45:35 2006 22, 16 -- Received: from mitconexch.mitcon.org (mitconexch.mitcon.org [192.251.56.2]) 22, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JLjYkp003923 22, 16 -- for {microscopy-at-microscopy.com} ; Wed, 19 Jul 2006 16:45:34 -0500 22, 16 -- Received: by mitconexch.mitcon.org with Internet Mail Service (5.5.2656.59) 22, 16 -- id {M0R0NM63} ; Wed, 19 Jul 2006 17:45:33 -0400 22, 16 -- Message-ID: {089C7709BE9235448E3622C6F38D828F03EF6010-at-mitconexch.mitcon.org} 22, 16 -- From: "Battjes, Kevin" {battjes-at-impactanalytical.com} 22, 16 -- To: "'TindallR-at-missouri.edu'" {TindallR-at-missouri.edu} , 22, 16 -- "Microscopy List (E-mail)" {microscopy-at-microscopy.com} 22, 16 -- Subject: RE: [Microscopy] LKB Knifemaker on the skids 22, 16 -- Date: Wed, 19 Jul 2006 17:45:29 -0400 22, 16 -- MIME-Version: 1.0 22, 16 -- X-Mailer: Internet Mail Service (5.5.2656.59) 22, 16 -- Content-Type: text/plain; 22, 16 -- charset="iso-8859-1" ==============================End of - Headers==============================
On Jul 18, 2006, at 6:14 PM, gary-at-gaugler.com wrote:
} Does anyone know of an alternative to the Haskris R050 air cooled } chiller for the LEO/Zeiss EMs? The Haskris unit I have is failing at } least every year. The pump/motor unit will seize. Then the } compressor control circuitry will fry. } } Zeiss put special criteria on the chiller--45PSI, 30GPH, 72F. I } don't see any other maker that can match this. } Dear Gary, Have you looked into the water-cooled Haskris units? Our two have had no problems since installation ~3 years ago, and the (larger) Haskris units on the HVEM at Albany NY gave virtually trouble-free performance for more than 20 years. We do and did regular preventive maintenance--checking flow rates, changing filters, adjusting pH anti-algae and anti-corrosion additives, etc. Here at Caltech, we have a house chilled water supply for cooling. Do you have enough air flow to cool your units properly, or do the specs indicate that you should use a larger unit? Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Wed Jul 19 17:13:33 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6JMDWPa014798 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 17:13:33 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 7065C109B07 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 15:13:32 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id C16F6355A6 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 19 Jul 2006 15:13:26 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200607190114.k6J1EXpI029214-at-ns.microscopy.com} 4, 22 -- References: {200607190114.k6J1EXpI029214-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {56d29ad5847931b087bff96111bbacad-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] 4, 22 -- Date: Wed, 19 Jul 2006 15:14:55 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Be sure to get a new Freon filter for your installation. Tecumseh makes the compressor and evaporator units. Current model is AKA4460YXD and should come with the current relay and start capacitor.
gary g.
At 01:28 PM 7/19/2006, you wrote: } Gary, } } We're in the process of replacing compressor and condenser unit because } this second compressor has worn out. We are changing to modern } refrigerant at the same time, hence the new condenser unit. Haskris } will tell you what model to get from local suppliers. You don't have to } buy it from them. The whole unit costs about $700. } } Ron L } } -----Original Message----- } From: gary-at-gaugler.com [mailto:gary-at-gaugler.com] } Sent: Wednesday, July 19, 2006 4:21 PM } To: lherault-at-bu.edu } Subject: [Microscopy] Re: Haskris chiller for Zeiss/LEO } } } } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America
We've been looking at fungal hyphae in plant root tissues, and they contain unidentified compounds in what look like lipidic vesicles (ranging in size from tiny to a few microns in diameter) that are fluorescent under UV. After staining tissue plus fungus with toluidine blue, the compounds are substantially more fluorescent - blue-white fluorescence, which goes against any ancestral knowledge I've heard about - I usually use tol. blue to quench unwanted autofluorescence. Not only that, but the vesicle fluorescence increases substantially with increasing exposure to UV.
Any ideas as to what type of compound(s) these might be? Something with ring structures to absorb light - does animal lipofuchsin respond this way to UV?
thanks for any suggestions, cheers, Rosemray
Dr. Rosemary White rosemary.white-at-csiro.au CSIRO Plant Industry ph. 61 (0)2-6246 5475 GPO Box 1600 mob. 61 (0)402 835 973 Canberra, ACT 2601 fax. 61 (0)2-6246 5334 Australia
we have the exact same knife maker and the schematics + service instructions. There are only about six pages so I can easily scan them as JPEGs and email them off-list if you wish.
My only concern is that you should never lubricate the flat surfaces between the cutting head and the main stand with the handle. These rely on metal/metal contact.
If the head is no longer tightening at the right height there is a simple adjustment which involves removing a domed chrome screw at the front of the cutter stand near the clamping handle. Beneath this is a 3mm hexagonal screw which can be loosened slightly. When this is done you should be able to rotate a collar at the base of the clamping handle. This adjusts the position at which the locking lever clamps - so just try turning it a bit and test locking arm until it clamps at about the horizontal position. Then re-tighten the 3mm hexagonal screw and put the covering chrome screw back.
I can also send the schematics if you haven't already got them - LET ME KNOW, THOUGH. These details are under IV Troubleshooting Fault 6.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: TindallR-at-missouri.edu
Dear group: - any chance someone out there has a copy of the operating manual for this model of centrifuge - it is refrigerated. Thanks Barbara
==============================Original Headers============================== 1, 18 -- From maloneyb-at-fiu.edu Thu Jul 20 10:18:27 2006 1, 18 -- Received: from smtp8.fiu.edu (smtp8.fiu.edu [131.94.79.25]) 1, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KFIQiU008830 1, 18 -- for {microscopy-at-msa.microscopy.com} ; Thu, 20 Jul 2006 10:18:26 -0500 1, 18 -- Received: from [131.94.95.156] (esga680.fiu.edu [131.94.95.156]) 1, 18 -- by smtp8.fiu.edu (MOS 3.7.1-GA) 1, 18 -- with ESMTP id CAI24841; 1, 18 -- Thu, 20 Jul 2006 11:18:23 -0400 (EDT) 1, 18 -- Message-ID: {44BF9F01.4020605-at-fiu.edu} 1, 18 -- Date: Thu, 20 Jul 2006 11:19:29 -0400 1, 18 -- From: Barbara Maloney {maloneyb-at-fiu.edu} 1, 18 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 1, 18 -- X-Accept-Language: en-us, en 1, 18 -- MIME-Version: 1.0 1, 18 -- To: microscopy-at-msa.microscopy.com 1, 18 -- Subject: Jouan CR4.11 centrifuge 1, 18 -- Content-Type: text/plain; charset=us-ascii; format=flowed 1, 18 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Hello, I think the internal safety pressure seal on our critical point drier has burst. Does anyone know where I can get a PELCO CPD2 Critical point drier repaired at?
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 2, 24 -- From gvrdolja-at-nature.berkeley.edu Thu Jul 20 11:29:44 2006 2, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 2, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KGTiaX020133 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 11:29:44 -0500 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 71648C1E8C 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 09:29:44 -0700 (PDT) 2, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 2, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 2, 24 -- with ESMTP id 17439-05 for {microscopy-at-microscopy.com} ; 2, 24 -- Thu, 20 Jul 2006 09:29:34 -0700 (PDT) 2, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 2, 24 -- id 496F8C1E9B; Thu, 20 Jul 2006 09:29:34 -0700 (PDT) 2, 24 -- Received: from localhost (localhost [127.0.0.1]) 2, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 40A37C1E8C 2, 24 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 09:29:34 -0700 (PDT) 2, 24 -- Date: Thu, 20 Jul 2006 09:29:34 -0700 (PDT) 2, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 2, 24 -- To: microscopy-at-microscopy.com 2, 24 -- Subject: repair of critical point drier 2, 24 -- Message-ID: {Pine.SOC.4.64.0607200927360.11284-at-nature.Berkeley.EDU} 2, 24 -- MIME-Version: 1.0 2, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 2, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 ==============================End of - Headers==============================
I don't know how well etching will work with this material. Somebody else on the listserver will probably know better than me.
Alternative, you can stain with ruthenium tetroxide and examine using SEM, FE-SEM or TEM. Procedure is well described in following reference. This should be no problem if the components can be stained with RuO4.
GM Brown and JH Butler, New method for the characterization of domain morphology of polymer blends using ruthenium tetroxide staining and low voltage scanning electron microscopy (LVSEM), Polymer 38 (15), 3937-3945, 1997.
Regards,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Sadhukhan, Pat" {SadhukhanPat-at- To BFUSA.com} {gary.m.brown-at-exxonmobil.com} cc
Dear Gary: Can we etch rubber particles from a cross-linked rubber/plastic composite ? If so, can you kindly suggest a procedure ? Regards, Pat Sadhukhan
Igauthier,
I expect that you will be disappointed. Typically, elucidation of the morphology of crystalline morphology of polymers generally requires etching or staining.
Regards,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
==============================Original Headers============================== 26, 19 -- From gary.m.brown-at-exxonmobil.com Thu Jul 20 12:21:03 2006 26, 19 -- Received: from hoespc02.exxonmobil.com (hoespc02.exxonmobil.com [192.67.48.39]) 26, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KHL3q6031222 26, 19 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 12:21:03 -0500 26, 19 -- Received: from hounmg03.NA.XOM.COM (hounmg03.na.xom.com [158.35.101.41]) 26, 19 -- by hoespc02.exxonmobil.com (Switch-3.1.7/Switch-3.1.7) with ESMTP id k6KHL0Ob011027 26, 19 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 12:21:01 -0500 (CDT) 26, 19 -- In-Reply-To: {1178C664B84FD311AE9B0008C7A4F0BF05627D4E-at-serverex.bfs.com} 26, 19 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: Observation of Polymers morphology 26, 19 -- Importance: 26, 19 -- To: "Sadhukhan, Pat" {SadhukhanPat-at-BFUSA.com} , microscopy-at-microscopy.com 26, 19 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 26, 19 -- Message-ID: {OFEE8F83CD.0BCAA994-ON862571B1.005EBB62-862571B1.005F4D3C-at-exxonmobil.com} 26, 19 -- From: gary.m.brown-at-exxonmobil.com 26, 19 -- Date: Thu, 20 Jul 2006 12:20:58 -0500 26, 19 -- X-MIMETrack: Serialize by Router on Hounmg03.na.xom.com/S/ExxonMobil(652FP1HF193|March 26, 19 -- 02, 2006) at 07/20/2006 12:21:01 PM 26, 19 -- MIME-Version: 1.0 26, 19 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (dfine-at-seton.org) from on Thursday, July 20, 2006 at 13:24:06 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both dfine-at-seton.org as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: dfine-at-seton.org Name: Diann Fine
Organization: Brackenridge Hospital
Education: Graduate College
Location: Austin, Texas, U.S.A.
Title: Tissue Processing for EM
Question: We are considering purchasing a Lynx el tissue processor from Electron Microscopy Sciences. I would appreciate some opinions, pro and con concerning the Lynx. Also, we are looking at obtaining a vacuum oven from EMS, either the hydraulic thermostat controlled or the programmable microprocessor temperature controlled. Thank you all for your help.
We have routinely used the Lynx el Processor for approximately 15 years now. Extremely reliable. We have an established 14-vial processing protocol following fixation and rinsing of minced tissues: rinse, osmication, dehydration, PO, infiltration through pure resin - -~15ml each. Max time is 26 hours, though it's set to run overnight so whether you start at 8AM or 4PM, it's ready the next morning.
We routinely use the 4-well baskets. To maximize infiltration, don't overload wells. As we only embed 5 samples per tissue, I only include 5-6 minced pieces. On rare occasions, a sample will work it's way through the slots, but they never have crossed into other vials.
Cleaning is with lots of acetone. Osmium will react with metallic stem pieces and thus we rinse in acetone, then light wipe/scrub with Soft Scrub, and final sonication in diluted detergent.
The system is designed to process 7baskets x 4 wells each, but I typically will limit myself to 5 baskets (20 unique tissue samples) at a time. 3-well and 8-well baskets are also available.
The only other similar unit is from Leica at http://www.leica-microsystems.com/EM_Specimen_Prep
Best regards. Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"The ultimate human freedom is the ability to choose one's attitude in a given set of circumstances." -Viktor Frankl
-----Original Message----- X-from: dfine-at-seton.org [mailto:dfine-at-seton.org] Sent: Thursday, July 20, 2006 2:58 PM To: Bobrowski, Walter
This Question was submitted to Ask-A-Microscopist by (dfine-at-seton.org) from on Thursday, July 20, 2006 at 13:24:06 Remember to consider the Grade/Age of the student when considering the Question ------------------------------------------------------------------------ --- Please reply to both dfine-at-seton.org as well as to the Microscopy Listserver ------------------------------------------------------------------------ ---
Email: dfine-at-seton.org Name: Diann Fine
Organization: Brackenridge Hospital
Education: Graduate College
Location: Austin, Texas, U.S.A.
Title: Tissue Processing for EM
Question: We are considering purchasing a Lynx el tissue processor from Electron Microscopy Sciences. I would appreciate some opinions, pro and con concerning the Lynx. Also, we are looking at obtaining a vacuum oven from EMS, either the hydraulic thermostat controlled or the programmable microprocessor temperature controlled. Thank you all for your help.
==============================Original Headers============================== 8, 12 -- From zaluzec-at-microscopy.com Thu Jul 20 13:51:20 2006 8, 12 -- Received: from [10.255.254.48] (msdvpn26.msd.anl.gov [130.202.238.90]) 8, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KIpIb9010602 8, 12 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 13:51:19 -0500 8, 12 -- Mime-Version: 1.0 8, 12 -- X-Sender: (Unverified) 8, 12 -- Message-Id: {p06020407c0e58118a310-at-[10.255.254.48]} 8, 12 -- Date: Thu, 20 Jul 2006 14:51:22 -0400 8, 12 -- To: microscopy-at-microscopy.com 8, 12 -- From: dfine-at-seton.org (by way of Ask-A-Microscopist) 8, 12 -- Subject: AskAMicroscopist: Tissue Processing for EM 8, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers============================== ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately.
==============================Original Headers============================== 26, 29 -- From Walter.Bobrowski-at-pfizer.com Thu Jul 20 14:42:56 2006 26, 29 -- Received: from gromsgoa01.pfizer.com (gromsgo.pfizer.com [148.168.34.153]) 26, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KJguIF021850 26, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 14:42:56 -0500 26, 29 -- Received: from groamrexc01.amer.pfizer.com (groamrexc01.amer.pfizer.com [172.30.8.168]) 26, 29 -- by gromsgoa01.pfizer.com (8.13.4/8.13.4) with ESMTP id k6KJdwA6006752; 26, 29 -- Thu, 20 Jul 2006 15:39:58 -0400 26, 29 -- Received: from groamrexc02.amer.pfizer.com ([172.30.8.169]) by groamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.211); 26, 29 -- Thu, 20 Jul 2006 15:42:53 -0400 26, 29 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by groamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.211); 26, 29 -- Thu, 20 Jul 2006 15:42:53 -0400 26, 29 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.7226.0 26, 29 -- Content-class: urn:content-classes:message 26, 29 -- MIME-Version: 1.0 26, 29 -- Content-Type: text/plain; 26, 29 -- charset="us-ascii" 26, 29 -- Subject: RE: [Microscopy] AskAMicroscopist: Tissue Processing for EM 26, 29 -- Date: Thu, 20 Jul 2006 15:42:31 -0400 26, 29 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D07407EBB-at-anaamrexm01.amer.pfizer.com} 26, 29 -- X-MS-Has-Attach: 26, 29 -- X-MS-TNEF-Correlator: 26, 29 -- Thread-Topic: [Microscopy] AskAMicroscopist: Tissue Processing for EM 26, 29 -- Thread-Index: AcasLmH+Jm0RptsvTWmGLL8i9w1EMgABEAxg 26, 29 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 26, 29 -- To: {dfine-at-seton.org} , {Microscopy-at-microscopy.com} 26, 29 -- X-OriginalArrivalTime: 20 Jul 2006 19:42:53.0606 (UTC) FILETIME=[B1181460:01C6AC34] 26, 29 -- X-Proofpoint-Spam-Reason: safe 26, 29 -- Content-Transfer-Encoding: 8bit 26, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6KJguIF021850 ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (dfine-at-seton.org) from on Thursday, July 20, 2006 at 15:05:16 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both dfine-at-seton.org as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: dfine-at-seton.org Name: Diann Fine
Organization: Brackenridge Hospital
Education: Graduate College
Location: Austin, Texas, USA
Title: Leica EMPT
Question: I would appreciate some information from the EM labs using the Leica EMTP. Is it user-friendly and reliable? Would you purchase it again? Were there any problems or issues with it? Do you run 2 different processing programs together at the same time? Do you run separate procedures for kidney and muscle biopsies? Are different procedures run for each different tissue? Thanks for the replies.
I was wondering what that slotted screw was for and even had it out, but didn't notice the hex screw underneath. The arm is now adjusted to lock just above horizontal.
It's still a little off, because I have to apply way too much pressure on the locking lever to keep it from riding up when the breaking wheel is turned. And that's a Bad Thing. But we are making knives and I'll keep playing with the fine-tuning.
Thanks for all the very informative responses.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Thu Jul 20 16:53:44 2006 10, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6KLrh6j012049 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 20 Jul 2006 16:53:43 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Thu, 20 Jul 2006 16:53:43 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: LKB Knifemaker 10, 23 -- Date: Thu, 20 Jul 2006 16:53:43 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E3394-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: LKB Knifemaker 10, 23 -- thread-index: AcasRve4gW1T67jgRWG66JuqMTWodg== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 20 Jul 2006 21:53:43.0476 (UTC) FILETIME=[F7FB2F40:01C6AC46] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6KLrh6j012049 ==============================End of - Headers==============================
That software would be BTVPro from http://www.bensoftware.com/ timelapse, motion detection, frame averaging, file export and many other features On Mar 1, 2005, at 8:04 AM, James Pawley wrote:
} } } ---------------------------------------------------------------------- } -------- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } --------- } } } --------------------------------------------------------------------- } } --------- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } --------------------------------------------------------------------- } } ---------- } } } } Roger; } } } } You might find some past discussions, or care to raise } } this issue again in the Yahoo Microscope group. Both } } issues (LED Illumination and inexpensive camera } } options) have been discussed there in the past. That } } group is more focused (no pun intended) on LM. It is } } also more of an amateur orientated group, although } } many of the prime contributors are professionals (or } } retirees). As a result of the audiance, a number of } } low cost but "workable" alternatives are discussed. } } } } The latest option for cameras seems to be using "Web } } Cams", either out of the box (with standard occulars), } } or by removing the lens assemblies and using a Photo } } or Projection eyepiece. } } } } I've played with LEDs myself, and have experienced } } results similar to what you describe (OK, but not } } spectacular). For me, it is more a matter of a light } } source that can be used "in the field". } } } } John Raffensperger, Jr. } } Beaver Dam, Wisconsin } } } } Hi all, } } I hope that it is not too commercial to tell you that, in the next } edition of The Handbook, there will be a fairly complete discussion } of how LEDs might be used in microscopy in the chapter on Non-laser } Light Sources by Andreas Nolte from Zeiss. At least it shows that } the idea has promise. } } And on the cheap CCD front, I have had fair success hooking the } iSight camera from apple (~$140, 640x480, Firewire) to a Zeiss } scope by the simple expedient of removing the rubber "eye-glasses" } protector gasket on the normal 10x high-eyepoint occular and } holding the camera right in front of it with a plastic collar. It } covers a little more than the whole field of view (some black } corners). Although I haven't used it, I am told that one can use } $30 "surveillance" software to record time-lapse sequences on a Mac } using this camera. Might be neat for non-microscope uses too. } } Cheers, } } Jim P. } -- } ********************************************** } Prof. James B. Pawley, Ph. } 608-263-3147 Room 223, Zoology Research Building, } FAX 608-265-5315 } 1117 Johnson Ave., Madison, WI, 53706 JBPAWLEY-at-WISC.EDU } 3D Microscopy of Living Cells Course, June 11-23, 2005, UBC, } Vancouver Canada } Info: http:// www.3dcourse.ubc.ca/ Applications due by March } 15, 2005 } } Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
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I have put my answers alonside your questions.
} Email: dfine-at-seton.org } Name: Diann Fine } } Organization: Brackenridge Hospital } } Education: Graduate College } } Location: Austin, Texas, USA } } Title: Leica EMPT } } Question: I would appreciate some information from the EM labs using the } Leica EMTP. Is it user-friendly and reliable?
} YES - on both counts, based on our experience of using it for a year for } acrylic and epoxy resin processing for high res LM and TEM respectively.
} Would you purchase it again?
} YES - in my view it is by far the best processor on the market for dual } purpose high throughput LM resin histology and TEM
} Were there any problems or issues with it?
} YES - the turntable height had to be adjusted to a lower position as the } vials were catching on the bottom of the cooling jacket. This has been the } only service callout in the year we have been using it.
} Do you run 2 different processing programs together at the same time?
} I don't think that it is possible to do this.
} Do you run separate procedures for kidney and muscle biopsies? YES
} Are different procedures run for each different tissue? NO
We have 2 basic schedules for both epoxy and acrylic resin processing. A relatively short (working day) schedule for small, soft easy to infiltrate tissues such as renal biopsies and longer (overnight) schedule for muscle and other tissues that are more difficult to infiltrate.
} Thanks for the replies.
Get back to me if you want a copy of our protocols.
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Email: nessonm-at-onid.orst.edu Name: Michael Nesson
Organization: Oregon State University EM Facility
Title-Subject: [Filtered] X-ray Detector Repair
Question: We have a blown window on our Kevex detector on our FEI CM12 TEM. Please respond off-line to me if you have had either exceptionally good or exceptionally bad experiences with the three companies that advertise in "Microscopy Today": namely Advanced Analysis Technologies, e2v Scientific Instruments (formerly Gresham), and MAX DETECTOR Repair Group LLC. Again, please respond to me directly (nessonm-at-onid.orst.edu), rather than to the Listserv, so as to reduce the possibility of flame wars. TIA, Mike Nesson
Michael H. Nesson, Ph.D. Electron Microscope Facility 1078 Cordley Hall Oregon State University Corvallis, OR 97331 (541) 737-5245
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Title-Subject: [Filtered] Re: [Microscopy] viaWWW: LKB 8800A-NM Ultrotome III Problem
Question: Following up on my recent LKB III Microtome troubles...
First, thank you to all that responded, your comments & suggestions were very helpful.
So, after investigating further this is what I have concluded: 1. The cord/string that raises the specimen arm is intact and functional. When I lift the arm up with my hand it moves as it should. Also, when I move the arm into "lock position", the arm is lifted up. 2. The belt in my control unit that is attached to the manual control knob is intact and moves as it should when I turn the knob. However, the specimen arm does not move. 3. There is no resistance when I move the manual control know, thus I don't think the potentiometer is seized up.
Also - when I switch to "auto" mode using the manual control knob, nothing happens. There is still a connection with the control unit however as the lamp still turns on.
Any further suggestions would be greatly appreciated!
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Email: javaidqazi-at-kemet.com Name: javaid qazi
Title-Subject: [Filtered] microscopy training
Question: Apart from Lehigh is there any other place which does training for SEM users.
The College of Microscopy located in Westmont IL offers SEM courses twice a year. Check out their website
www.collegeofmicroscopy.com
Regards
Alan
At 10:24 AM 7/21/2006 -0500, javaidqazi-at-kemet.com wrote:
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Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
Yes, the McCrone group in Westmount, Il, outside of Chicago. They're nice people.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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javaidqazi-at-kemet. com To: frank.karl-at-degussa.com cc: 07/21/2006 11:24 Subject: [Microscopy] viaWWW: microscopy training AM Please respond to javaidqazi
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Email: javaidqazi-at-kemet.com Name: javaid qazi
Title-Subject: [Filtered] microscopy training
Question: Apart from Lehigh is there any other place which does training for SEM users.
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Email: hu.duan-at-averydennison.com Name: Hu
Title-Subject: [Filtered] question regarding roughness in large area
Question: Dear colleagues:
I have an application to quantitatively monitor the roughness/topology change over a relatively large area (3-5cm*3-5cm). The surface is not uniform. Thus, monitoring a large area is necessary. The conventional AFM, optical profilometry techniques suffer due to their limited scanned areas. Even with image stitching, the results are not satisfied due to complication of tilt, plane fit over such large area. I am wondering is there such technique/instrumentation to do such kind of work? Also I know mechanical profiler like Dektak can do similar work. However, our coating is relatively soft, usual stylus applies too much force and often creates scratches. Are there any optical methods for such application? Thank you for your attention and suggestions.
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Email: Sandow50-at-yahoo.com Name: Stanton Dowd
Organization: VCU
Title-Subject: [Filtered] Deton DV-502A High Vac issues
Question: I have been using an old DV-502A for routine carbon coating for the past four years without any significant problems, but recently I have had problems getting it down to operating vacuum (~1x10^-4) without using liquid nitrogen in the trap. Until about a month ago I've never needed to use the trap to achieve a much stronger vacuum. The problem seems to be getting worse without any overt reason. I've changed out the mechanical pump and damn near every seal in the thing in the past few weeks with no effect. As far as I can tell there is no significant leak anywhere, it simply seems that the diffusion pump efficiency has tanked. Using nitrogen in the trap I can still pump down to well under 1x10^-5 Torr, but without it the diffusion pump barely works at all. Assuming that there are no leaks, that the mechanical pump is working as it should, that the diffusion oil is clean and at the appropriate level and that the tree is intact; what can effect diffusion pumping efficiency? What I am left to wonder about is the column temperature gradient. I have the model with the 3.5" column if that matters. The peak temp is about one inch from the bas a reads at around 365C. The temp drops quickly over the first three inches and reads around 85C at the first cooling coil. There is a smooth gradient up the column to the top where the temp is right at 30C. I never bothered to take readings when it was working so I don't know if these numbers are normal or not. We are using tap water as the coolant and draining it into the sink. With the weather as warm as it has been I wonder if the influent temp is too warm to cool it adequately. For what its worth, the influent is 25.5C with a flow rate of 1.65 L/min and an effluent temp of 28C.
Any help that you guys can give would be greatly appreciated.
Water T is good.You don't want it colder than 15C and hotter than 30C (at supply end), optimum at return end is between 30 and 35C. Actual numbers may depend on flow rate, but it is good to stay within above limits. Speaking of leaks- TC (thermocouple) sensor reads below 20 mTorr with main valve open, there is no leaks. High vacuum leak always results in elevated backing pressure of the diff. pump. No elevated back pressure means no leak. I assume that mech. pump works well, which is tested by pumping lines only with all valves closed.
The potential diff. pump problems are:
1) Denton diff. pumps have water cooling tube coil cemented to the pump with heat conductive cement. More classy and expensive pumps such as Edwards or Varian have cooling tubing welded to pump column. Cement eventually cracks, and stops transferring heat. You need to examine it very closely and try to move the water cooling coil to see if it is still bonded to the pump. If it isn't, then either re-cement, or take pump off the unit, disassemble, clean it, and braze or solder water tubing to the pump column.
2) Depleted or lost diff. pump fluid. Check and replace or re-fill as necessary.
3) Silicone diff. pump fluid (such as Dow Corning 702 or 704 or 705) that is typically used by Denton may partially polymerize inside the pump. It will look clear and otherwise normal, with one difference. Touch the cooling tower inside the pump, and you will feel sticky layer - feels like sticky packaging tape or note stickers from Office Depot. Layer is clear and invisible, but easily detectable by touch. If this happened, pump is dead, even though fluid level is normal and all looks clean. Discard old fluid, disassemble the pump, and clean in very diligently. Acetone rub works for sticky stuff, but solid deposits (if present) must be removed with wire brush or sandblasting.
Vitaly Feingold SIA 2773 Heath Line Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com ----- Original Message ----- X-from: {Sandow50-at-yahoo.com} To: {vitalylazar-at-att.net} Sent: Monday, July 24, 2006 8:33 PM
Stanton,
Remove the DP and check that the DP oil is good. It might be 'cracked'. Replace it if you need to do that. I would do it anyway.
We had a very similar experience to yours on a JSM 35 SEM. The factory spent 3 weeks looking for a vacuum leak with a helium leak detector and examined every o-ring in the system. No good. A different serviceman came in and found the problem in one hour. The one terminal connection on a three month old DP heater was corroded. It was installed by the first serviceman to solve the same problem months ago. A new heater and terminal connection restored the pumping capacity to normal. Apparently the terminal limited the current to the DP heater, the DP got hot, but it never got hot enough to restore full vacuum pumping speed. LN2 helped but we knew the non-LN2 vacuum was bad. This seems to be a carbon copy of your problem and your assumptions of what is working properly.
You asked, "What can effect diffusion pumping efficiency?" The DP chimney port is not lined up with the rotary pump port. The DP oil is cracked. The DP oil is less than normal. The DP heater is defective in some way. Insufficient DP heating. A bad DP heater connection. Hot or limited cooling water. The "loss of cooling water" safety switch on the DP is opening at a lower temp. Cracked or leaking o-ring at the DP interface with the LN2 trap.
Paul
} Email: Sandow50-at-yahoo.com } Name: Stanton Dowd } } Organization: VCU } } Title-Subject: [Filtered] Deton DV-502A High Vac issues } } Question: I have been using an old DV-502A for routine carbon coating for the past four years without any significant problems, but recently I have had problems getting it down to operating vacuum (~1x10^-4) without using liquid nitrogen in the trap. Until about a month ago I've never needed to use the trap to achieve a much stronger vacuum. The problem seems to be getting worse without any overt reason. I've changed out the mechanical pump and damn near every seal in the thing in the past few weeks with no effect. As far as I can tell there is no significant leak anywhere, it simply seems that the diffusion pump efficiency has tanked. Using nitrogen in the trap I can still pump down to well under 1x10^-5 Torr, but without it the diffusion pump barely works at all. } Assuming that there are no leaks, that the mechanical pump is working as it should, that the diffusion oil is clean and at the appropriate level and that the tree is intact; what can effect diffusion pumping efficiency? What I am left to wonder about is the column temperature gradient. I have the model with the 3.5" column if that matters. The peak temp is about one inch from the bas a reads at around 365C. The temp drops quickly over the first three inches and reads around 85C at the first cooling coil. There is a smooth gradient up the column to the top where the temp is right at 30C. I never bothered to take readings when it was working so I don't know if these numbers are normal or not. We are using tap water as the coolant and draining it into the sink. With the weather as warm as it has been I wonder if the influent temp is too warm to cool it adequately. For what its worth, the influent is 25.5C with a flow rate of 1.65 L/min and an effluent temp of 28C. } } Any help that you guys can give would be greatly appreciated. } } Sincerely, } Stanton }
==============================Original Headers============================== 8, 26 -- From beaurega-at-westol.com Tue Jul 25 07:51:27 2006 8, 26 -- Received: from smtp-gateway-6.winbeam.com (smtp-gateway-6.winbeam.com [64.84.97.71]) 8, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6PCpRx2022896 8, 26 -- for {microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 07:51:27 -0500 8, 26 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 8, 26 -- by smtp-gateway-6.winbeam.com (8.13.1/8.12.8) with SMTP id k6PCpKTj013505 8, 26 -- for {microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 08:51:21 -0400 8, 26 -- Received: (qmail 22938 invoked by uid 89); 25 Jul 2006 12:51:09 -0000 8, 26 -- Received: from pitts-69-72-13-84.dynamic-dialup.coretel.net (HELO millenium) (69.72.13.84) 8, 26 -- by mail.winbeam.com with SMTP; 25 Jul 2006 12:51:09 -0000 8, 26 -- Message-Id: {3.0.6.32.20060725085246.007d7560-at-pop3.norton.antivirus} 8, 26 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus (Unverified) 8, 26 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 8, 26 -- Date: Tue, 25 Jul 2006 08:52:46 -0400 8, 26 -- To: microscopy-at-microscopy.com 8, 26 -- From: Beaurega {beaurega-at-westol.com} 8, 26 -- Subject: Re: [Microscopy] Re: viaWWW: Deton DV-502A High Vac issues 8, 26 -- In-Reply-To: {200607250217.k6P2HALh003819-at-ns.microscopy.com} 8, 26 -- Mime-Version: 1.0 8, 26 -- Content-Type: text/plain; charset="us-ascii" 8, 26 -- X-Winbeam-MailScanner-Information: smtp-gateway-6.winbeam.com - Please contact Technical Support for more information 8, 26 -- X-Winbeam-MailScanner: Found to be clean [smtp-gateway-6.winbeam.com] (courtesy of Winbeam) 8, 26 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, 8, 26 -- score=0.276, required 6, autolearn=disabled, 8, 26 -- MAILTO_TO_SPAM_ADDR 0.28) 8, 26 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
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Email: khibbs-at-wistar.org Name: Kristine Hibbs
Organization: The Wistar Institute
Title-Subject: [Filtered] Announcement of Job Opening
Question: POSITION IN MICROSCOPY
THE WISTAR INSTITUTE
Research Assistant
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The Wistar Institute has an opening for a Research Assistant who will manage the multi (two)-photon microscope at the Institute. The individual will operate the microscope and interact with users. BachelorÇs degree required, but MasterÇs degree preferred. Background in microscopy required. Experience with confocal laser and/or two-photon microscopy required. Previous lab experience desired. Ability to problem solve and work independently required.
We offer an excellent benefits package, including tuition assistance. We do not offer relocation assistance. To apply, visit our webpage at http://www.wistar.org/humanresources/employment.htm and submit your resume and cover letter stating salary requirements and previous experience with confocal laser and/or two-photon microscopy.
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For more information about The Wistar Institute visit our website at www.wistar.org.
Kristine Hibbs Human Resources Specialist The Wistar Institute (215) 898-3766 Khibbs-at-wistar.org
I am trying to get some paraformaldehyde into solution. I have probably done this several thousand times in my career without a problem. I am using a fairly new bottle of paraformaldehyde (prill type) and putting 2% in water (deionized or distilled - both have the same problem). I heat to 55 C (not over 60 C) with vigorous stirring, add a few drops of 1 N NaOH and the bulk goes into solution. The problem is there is a significant amount of flocculent material which doesn't go into solution. I made up a fresh batch of NaOH and there was no improvement. Has anyone experienced a similar problem?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
As suggested by others, Stanton's problem with his diffusion pump may very well be due to insufficient heat input from the DP's heater. If you look on the manufacturer's tag that is probably attached to the pump somewhere, you should find the value of the heater input in Watts. Contact your local electrician and he should have a gadget that he can simply clip around the wire leading into the heater and measure the current flowing through it. Current (Amps) x voltage (volts) = Watts. If the value measured doesn't equal the value on the pump's tag, you know you have a problem with the heater. Then, disconnect the heater wires and measure the resistance of the heating elements to see if one is burned out. Making a few measurements like this should give you an indication of whether or not there is a problem with the heater. If not, check the other factors mentioned. Good luck, WCB -- Wilbur C. Bigelow, Professor Emeritus Materials Sci. & Engr., Univ. of Michigan Ann Arbor, Michigan 48109-2136 e-mail: bigelow-at-engin.umich.edu; Fx:734-763-4788; Ph:734-975-0858 Address mail to: 2911 Whittier Court Ann Arbor, MI 48104-6731
==============================Original Headers============================== 1, 14 -- From bigelow-at-engin.umich.edu Tue Jul 25 11:07:33 2006 1, 14 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6PG7Xd7025083 1, 14 -- for {microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 11:07:33 -0500 1, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 1, 14 -- by srvr22.engin.umich.edu (8.13.6/8.13.6) with ESMTP id k6PG7WRg018471 1, 14 -- for {microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 12:07:32 -0400 (EDT) 1, 14 -- Mime-Version: 1.0 1, 14 -- Message-Id: {p06210200c0ebef5a2b80-at-[141.212.131.221]} 1, 14 -- Date: Tue, 25 Jul 2006 12:07:31 -0400 1, 14 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 1, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 1, 14 -- Subject: [Microscopy}RE: Diff. Pump Problem 1, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Hello everybody, I used JB-4 resin for embedding of fish larvae. Tissue was fixed in 2% glut, dehydrated in ethanol and infiltrated with ethanol/resin mixture (50-50, 75-25, 100). Last infiltration was overnight. The resin was polymerized at RT under a vacuum. Blocks seem to be hard but I can not cut them on glass knife (I cut 3 micron section). The resin either curls up and/or sticks to the surface of a knife. I stored JB-4 at +4C, but it was brought to a RT before infiltration and embedding. I noticed that one of the components (accelerator) expired May 06. Do you have any suggestions/advice how to improve the procedure so the blocks can be cut? Thanks Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Tue Jul 25 13:10:59 2006 1, 22 -- Received: from mx.upei.ca (humboldt.cs.upei.ca [137.149.3.10]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6PIAxVm005474 1, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Jul 2006 13:10:59 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx.upei.ca with esmtp (Exim 3.35 #1 (Debian)) 1, 22 -- id 1G5RMw-0001VW-02 1, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Jul 2006 15:10:58 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 25 Jul 06 15:10:58 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 25 Jul 06 15:10:43 -0300 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-msa.microscopy.com 1, 22 -- Date: Tue, 25 Jul 2006 14:57:49 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: JB-4 resin problems 1, 22 -- Message-ID: {44C6316C.19011.108EB5-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
} As suggested by others, Stanton's problem with his diffusion } pump may very well be due to insufficient heat input from the } DP's heater. ...
I remember a similar problem, which could have well been the heater output ... But in this case it was because I had changed the DP fluid to another that had a higher boiling pt. The heater did work ... But it sure was sluggish in the beginning.
Genuinely, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre Memorial University St. John's, NL
==============================Original Headers============================== 7, 21 -- From michael-at-shaffer.net Tue Jul 25 13:39:13 2006 7, 21 -- Received: from ws6-5.us4.outblaze.com (ws6-5.us4.outblaze.com [205.158.62.152]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k6PIdCN0016239 7, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 25 Jul 2006 13:39:12 -0500 7, 21 -- Received: (qmail 24459 invoked from network); 25 Jul 2006 18:39:13 -0000 7, 21 -- Received: from unknown (HELO rarewolf) (michael-at-shaffer.net-at-205.251.84.119) 7, 21 -- by ws6-5.us4.outblaze.com with SMTP; 25 Jul 2006 18:39:12 -0000 7, 21 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 21 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 21 -- Cc: {bigelow-at-engin.umich.edu} 7, 21 -- Subject: RE: [Microscopy] [Microscopy}RE: Diff. Pump Problem 7, 21 -- Date: Tue, 25 Jul 2006 16:09:12 -0230 7, 21 -- Message-ID: {000b01c6b019$a08639e0$4701a8c0-at-rarewolf} 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: Microsoft Office Outlook 11 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 7, 21 -- Thread-Index: AcawBILzsLXAAdW2Qhi/oRSyFMrupwAFIf3g 7, 21 -- In-Reply-To: {200607251608.k6PG81A3025848-at-ns.microscopy.com} ==============================End of - Headers==============================
If you are heading to Chicago for the M&M meeting and you have an interest in High Pressure Freezing you are cordially invited to attend the following special events:
1. Sunday, July 30, 12:00 - 5:00pm, High Pressure Freezer User Group Meeting. Location: Sheraton Chicago Hotel & Towers, 301 East North Water St, Parlor E Buffet lunch served at 12:00 noon followed by a talk on High Pressure Freezing Techniques & Methods by Dr. Andres Kaech, Life Science Applications Manager, Bal-Tec AG at 1:00pm. Following this presentation the floor will be open for group discussion, questions & answers and/or individual presentations if anyone has a special topic to present to the group.
2. Thursday/Friday, August 3/4, small group High Pressure Freezing Workshop at the University of Chicago. This "hands-on" workshop hosted by the Dept of Molecular Genetics and Cell Biology and the Bioscience Division will be conducted in the laboratory of Dr. Joe Austin utilizing the Departments Bal-Tec HPM 010 High Pressure Freezer. Location: Lectures at 9:30am Thursday in CIS 400B (Center for Integrative Science, 4th floor) 929 E. 57th St (corner of 57th and Drexel) followed by lunch. Practical demonstrations and freezing in the lab, CIS ESB 06 (basement) in the afternoon and Friday morning starting at 9:00am The University of Chicago campus is approximately 25 minutes by car from the Navy Pier area. Please go to http://maps.uchicago.edu/westquad/irb.html to view a map of the building and nearby parking.
There is no charge to attend either of these meetings and lunch/refreshments are compliments of Bal-Tec RMC
See you in Chicago.
Dave Roberts Bal-Tec RMC Boeckeler Instruments Inc 4650 S. Butterfield Drive Tucson, AZ 85714 Tel: 520-745-0001 Fax: 520-745-0004 www.baltec-rmc.com
==============================Original Headers============================== 10, 20 -- From dave-at-boeckeler.com Tue Jul 25 16:12:04 2006 10, 20 -- Received: from txslsmtp2.vzwmail.net (txslsmtp2.vzwmail.net [66.174.85.156] (may be forged)) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6PLC4fG029472 10, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Jul 2006 16:12:04 -0500 10, 20 -- Received: from tweedledee (smtp.vzwmail.net [66.174.85.25]) 10, 20 -- (authenticated bits=0) 10, 20 -- by txslsmtp2.vzwmail.net (8.12.9/8.12.9) with ESMTP id k6PLBpxF016056 10, 20 -- for {microscopy-at-msa.microscopy.com} ; Tue, 25 Jul 2006 21:12:01 GMT 10, 20 -- Message-Id: {200607252112.k6PLBpxF016056-at-txslsmtp2.vzwmail.net} 10, 20 -- From: "Dave Roberts" {dave-at-boeckeler.com} 10, 20 -- To: {microscopy-at-msa.microscopy.com} 10, 20 -- Subject: High Pressure Freezing Events in Chicago 10, 20 -- Date: Tue, 25 Jul 2006 14:11:48 -0700 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; 10, 20 -- charset="US-ASCII" 10, 20 -- Content-Transfer-Encoding: 7bit 10, 20 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 10, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1409 10, 20 -- Thread-Index: AcawLvDyuWocdYaOST6Upc4KaWZBAA== ==============================End of - Headers==============================
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Email: martin.roe-at-nottingham.ac.uk Name: Martin Roe
Organization: Nottingham University
Title-Subject: [Filtered] JEOL 6000-series keyboard for 6400 SEM
Question: Dear Listservers, Does anyone have an old and spare JEOL 6000-series keyboard for a JEOL 6400 SEM (plugs into a DIN socket below the filament control)? Willing to pay, if necessary. Thanks Martin Roe
Martin Roe EM and XPS Technical Support Wolfson Building School of Mechanical, Materials, Manufacturing Engineering Nottingham University University Park Nottingham NG7 2RD England, UK
Yes, I have had a similar problem and found that I had to increase the temperature to 60 C, having found exactly as you describe at even a slightly lower temperature.
Given the hassle, I now by it in in sealed ampoules.
Hope this works for you,
Alastair
At 10:52 25/07/2006 -0500, you wrote:
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Alastair McKinnon Histology & EM Facility Manager University of Aberdeen, Institute of Medical Science Foresterhill, Aberdeen, AB25 2ZD tel: +44(0) 1224 552923 (office); +44(0) 1224 555911 (lab) fax: +44(0) 1224 559533; email: a.d.mckinnon-at-abdn.ac.uk www.abdn.ac.uk/ims/h-em
==============================Original Headers============================== 12, 18 -- From a.d.mckinnon-at-abdn.ac.uk Wed Jul 26 03:24:36 2006 12, 18 -- Received: from mailhub1.abdn.ac.uk (mailhub1.abdn.ac.uk [139.133.7.28]) 12, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6Q8OZwJ004959 12, 18 -- for {Microscopy-at-Microscopy.Com} ; Wed, 26 Jul 2006 03:24:35 -0500 12, 18 -- Received: from med-0069.ims.abdn.ac.uk ([139.133.159.90] helo=med-0069.abdn.ac.uk) 12, 18 -- by mailhub1.abdn.ac.uk with esmtp (Exim 4.52) 12, 18 -- id 1G5ehB-00018h-Tx; Wed, 26 Jul 2006 09:24:46 +0100 12, 18 -- Message-Id: {5.2.1.1.0.20060726091621.01151600-at-mailms.abdn.ac.uk} 12, 18 -- X-Sender: pat081-at-mailms.abdn.ac.uk 12, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 12, 18 -- Date: Wed, 26 Jul 2006 09:24:31 +0100 12, 18 -- To: phillipst-at-missouri.edu 12, 18 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 12, 18 -- Subject: Re: [Microscopy] paraformaldehyde problem 12, 18 -- Cc: Microscopy-at-Microscopy.Com 12, 18 -- In-Reply-To: {200607251552.k6PFqnwt023624-at-ns.microscopy.com} 12, 18 -- Mime-Version: 1.0 12, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
returned from an interesting conference(ULTRAPATH XIII, Rapid City, SD.,USA) and a short vacation, I have found your query concerning the making of paraformaldehyde stock solution.
I do not know about the source of your recipe for this, especially to heat the crude, hydrous solution with the PFA-powder ONLY unto "} 55 { degr.C (not above 60 degr.C)".
All the } old fashioned { sources about PFA-fixative I know (e.g. GEYER G. ed., Ultrahistochemie / Ultrahistochemistry G.FischerVerlag Stuttgart Germany 1973, or: PLATTNER H, Zingsheim HP. eds, Elektronenmikroskopische Methodik in der Zell- und Molekularbiologie, G.Fischer Verl. Stuttgart 1987, certainly as well as other [engl.] publications) state:
".....heat solution to approx. 65- } } 70 { { degr.C (but let NOT BOIL), - thoroughly mix up, add 1-3 drops [*whatever this means, depends perhaps on the dispensing device you use....*]
of 1 N NaOH......(and) until solution clears up....." [* I generally add drop after drop, waiting after each drop several minute wether the solution clears up or not*]
Perhaps the problem you face is a problem of incomplete dissolution due to a too low temperature.
IMO you would not need to discard such a solution, just filtering yields a fixative which perhaps has a slightly lower percentage in FA-concentration than you intended.
Another poblem would be precipitation of flocculent material after mixing the FA-solution with glutaraldehyde: this IMO points to a perhaps degraded or low quality batch of the concentrated GA-solution.
Greetings and best regards,
Wolfgang Muss SALZBURG, AUSTRIA
---------- Von: phillipst-at-missouri.edu[SMTP:phillipst-at-missouri.edu] Antwort an: phillipst-at-missouri.edu Gesendet: Dienstag, 25. Juli 2006 17:51 An: W.Muss-at-salk.at Betreff: [Microscopy] paraformaldehyde problem
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Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
The Biology Department at St. Francis Xavier University, seeks applications for a full-time Research Technician to interact with a group of ten research active, faculty and their students. The successful applicant should have a minimum of a B.Sc. preferably with experience operating laser confocal and electron microscopes. Experience in specimen preparation and image analysis are desired assets. The applicant must have a willingness to develop skills and instrumentation expertise in other areas, as required.
The Biology Department is housed in a newly-renovated building with microscope facilities that include a Philips EM410 TEM, a Jeol JSM 5300 SEM and an Olympus FV300 CLSM. The Biology Department has a general interest in aquatic biology, including invertebrate reproduction, vertebrate physiology, invasive algal and crustacean species, carbon fixation in cyanobacteria, marine algal symbiosis, as well as expertise in cell biology, microbiology and biomechanics.
This is a full time position with funding including benefits for 1 year initially followed by 2 additional years upon successful review of the applicant. Applications will be considered beginning August 15th, 2006 and the competition will remain open until the position is filled. Interested individuals should forward a CV outlining relevant experience, and the names and addresses of three references to:
Dr. J. Buckland-Nicks Chair, Department of Biology St. Francis Xavier University Antigonish, N.S. B2G 2W5 jbucklan-at-stfx.ca or mmurphy-at-stfx.ca
St. Francis Xavier University is committed to employment equity and welcomes applications from all qualified women and men, including aboriginal people, members of visible minorities, and persons with disabilities.
==============================Original Headers============================== 10, 20 -- From dmorriso-at-stfx.ca Wed Jul 26 07:37:27 2006 10, 20 -- Received: from xmail1.stfx.ca (xmail1.stfx.ca [141.109.221.24]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6QCbJDk029410 10, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 07:37:24 -0500 10, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 20 -- Content-class: urn:content-classes:message 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; 10, 20 -- charset="iso-8859-1" 10, 20 -- Subject: Job Posting: Research Technician in Microscopical Analysis 10, 20 -- Date: Wed, 26 Jul 2006 09:34:49 -0300 10, 20 -- Message-ID: {D1DA484CA5D7D5499AB6BBB4BCA4291750475F-at-demetrius.ad.stfx.ca} 10, 20 -- X-MS-Has-Attach: 10, 20 -- X-MS-TNEF-Correlator: 10, 20 -- Thread-Topic: Job Posting: Research Technician in Microscopical Analysis 10, 20 -- Thread-Index: Acawr+KzVfqA1H0FToqn46QL9Lr4nw== 10, 20 -- From: "Daniel Morrison" {dmorriso-at-stfx.ca} 10, 20 -- To: {Microscopy-at-microscopy.com} 10, 20 -- Content-Transfer-Encoding: 8bit 10, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6QCbJDk029410 ==============================End of - Headers==============================
Trilogy is a high pH antigen retrieval buffer used to unmask antigen sites in formalin fixed paraffin embedded tissue. It also has a detergent in it that can also deparaffinize tissue when used properly. I think it is an EDTA solution, pH 9-11?. As apposed to the normal Citrate buffer pH 6.0 that is normally used. Trilogy is only used when Citrate buffers and enzyme digestion, like trypsin, fail. We routinely used it in the clinical pathology lab on marker's like cd 10, cd 31, cd 4, cd 5, cd 8, etc.
"Effective unmasking, which reveals the antigen otherwise concealed by the negative effects of formalin fixation, has an undeniable and positive impact on the validation process. The mechanism of these negative effects is not yet completely understood but it is presumed that heating tissue sections in TrilogyTM causes the break up of the formalin cross-links that are believed to interfere with antigen-antibody binding. The discovery of the benefits of effective unmasking has been a revolutionary step in paving the road to standardization and the establishment of reliable validation procedures." From Cell Marque web site. They do not reveal their formula. =20
Hope this help's.
Rhonda Allen BA HT(ASCP)HTL, QIHC Histotechnology Specialist II Stowers Institute 1000 E. 50th Street Kansas City, MO 64110=20 816-926-4305
Rhonda Allen BA HT(ASCP)HTL, QIHC
==============================Original Headers============================== 13, 23 -- From rra-at-stowers-institute.org Wed Jul 26 09:44:20 2006 13, 23 -- Received: from stowers-institute.org (mail.stowers-institute.org [204.56.6.8]) 13, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6QEiKkY009517 13, 23 -- for {microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 09:44:20 -0500 13, 23 -- Received: from ([172.16.2.9]) 13, 23 -- by smtpkc01.stowers-institute.org with ESMTP id 4420336.1650497; 13, 23 -- Wed, 26 Jul 2006 09:43:50 -0500 13, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 23 -- Content-class: urn:content-classes:message 13, 23 -- MIME-Version: 1.0 13, 23 -- Content-Type: text/plain; 13, 23 -- charset="us-ascii" 13, 23 -- Subject: Trilogy 13, 23 -- Date: Wed, 26 Jul 2006 09:43:50 -0500 13, 23 -- Message-ID: {C28BAF593DC3314E9C0F3A50191C2E7804FB2300-at-EXCHKC03.stowers-institute.org} 13, 23 -- X-MS-Has-Attach: 13, 23 -- X-MS-TNEF-Correlator: 13, 23 -- Thread-Topic: Trilogy 13, 23 -- Thread-Index: AcawweiFHoky7NDoQ4yE/jGIQoPsPg== 13, 23 -- From: "Allen, Rhonda" {RRA-at-stowers-institute.org} 13, 23 -- To: {microscopy-at-microscopy.com} 13, 23 -- Content-Transfer-Encoding: 8bit 13, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k6QEiKkY009517 ==============================End of - Headers==============================
Postdoc (or professional programmer with scientific background) – Portland State University (Portland, Oregon/USA)
Initially for one year at $35,000 (to $38,000 depending on experience and background) plus 50 % fringe benefits (health insurance, retirement benefits, etc.), available from January 2nd, 2007, onwards, extendable up to 3 years by mutual agreement and if continuing funding over ONAMI {http://www.onami.us/} earmarks (or alternative funding agencies) can be secured.
The Nanocrystallography Group {http://www.physics.pdx.edu/%7Epmoeck/research%20group} at the Department of Physics {http://www.physics.pdx.edu/} of Portland State University {http://www.pdx.edu/} (PSU) is seeking a (male/female) postdoc (or professional programmer with scientific background) for a project on Image-based nanocrystallography with database support: Lattice-fringe fingerprinting to identify unknown nanocrystal phases & Nanocrystal morphology from tilt protocols {http://www.physics.pdx.edu/%7Epmoeck/projects/ONAMI-nanometrology/white%20paper%202006.htm} .
Excellent computer skills (e.g. C++, JScript, Java, Python, Perl, PHP, and MySQL) are essential. A background in crystallography, mineralogy, or materials science and engineering at the Masters (or PhD level) is desirable. Advanced degrees in computer science may substitute.
PSU is with an estimated enrollment of about 30,000 students Oregon’s largest and only urban university. It is located in downtown {http://maps.google.com/maps?oi=map&q=Portland,+OR} Portland/Oregon {http://www.portlandonline.com/} , i.e. the metropolis of the so called “silicon forest”. Since high tech industries blend in with abundant natural beauty in this region and because of its moderate climate, Portland has consistently been ranked in the top ten of “America’s most livable cities”.
The search will be open until the position has been filled. Applications (CV, list of referees, reasons for coming to the US for applicants from aboard, list of publications if applicable, etc.) should be sent to:
Prof. Peter Moeck {http://www.physics.pdx.edu/%7Epmoeck/}
Applications by email to pmoeck-at-pdx.edu {mailto:pmoeck-at-pdx.edu} with attachments as *.pdf files are acceptable. Since there is a hyperlink at the title of the project above, prospective candidates are advised to deliver a statement on how they might contribute to this multi-institutional collaborative effort.
-- Peter Moeck (PhD, Dr. rer. nat.) Assistant Professor
Portland State University, Department of Physics P.O. Box 751, Portland, OR 97207-0751 Tel.: 503 725 4227, Departmental fax.: 503 725 9525
pmoeck-at-pdx.edu
(Office 404, Science Building 2, 1719 SW 10th Ave, Portland, OR 97201)
http:/www.physics.pdx.edu/~pmoeck
==============================Original Headers============================== 21, 20 -- From pmoeck-at-pdx.edu Wed Jul 26 10:50:57 2006 21, 20 -- Received: from njord.oit.pdx.edu (njord.oit.pdx.edu [131.252.120.57]) 21, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6QFovaD020907 21, 20 -- for {Microscopy-at-Microscopy.Com} ; Wed, 26 Jul 2006 10:50:57 -0500 21, 20 -- Received: from pdx.edu (71-34-86-40.ptld.qwest.net [71.34.86.40]) 21, 20 -- (authenticated bits=0) 21, 20 -- by njord.oit.pdx.edu (8.13.3+/8.13.1) with ESMTP id k6QFos7e028578 21, 20 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 21, 20 -- for {Microscopy-at-Microscopy.Com} ; Wed, 26 Jul 2006 08:50:56 -0700 21, 20 -- X-Authentication-Warning: njord.oit.pdx.edu: Host 71-34-86-40.ptld.qwest.net [71.34.86.40] claimed to be pdx.edu 21, 20 -- Message-ID: {44C78F6C.4010904-at-pdx.edu} 21, 20 -- Date: Wed, 26 Jul 2006 08:51:08 -0700 21, 20 -- From: "Peter Moeck (Laptop - on the run)" {pmoeck-at-pdx.edu} 21, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 21, 20 -- X-Accept-Language: en-us, en 21, 20 -- MIME-Version: 1.0 21, 20 -- To: Microscopy-at-Microscopy.Com 21, 20 -- Subject: postdoc or professional programmer ($35k to $38k up to 3 years) 21, 20 -- Content-Type: text/plain; charset=windows-1252; format=flowed 21, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
MSc/PhD candidate positions in crystallography/applied physics
The Nanocrystallography Group {http://www.physics.pdx.edu/%7Epmoeck/research%20group} at the Department of Physics {http://www.physics.pdx.edu/} of Portland State University {http://www.pdx.edu/} is seeking candidates (m/f) for MSc and PhD projects in the field of transmission electron microscopy (TEM) based nanocrystallography. The graduate student projects focus on aspects of the development of novel nanocrystal characterization methods. These methods will utilize a combination of high-resolution phase contrast imaging in TEM and goniometry of direct lattice vectors {http://www.physics.pdx.edu/%7Epmoeck/goniometry.htm} .
The Nanocrystallography group is a user of Portland State University’s electron microscopy center {http://www.gatan.com/knowhow_8/psu_open_house.pdf} . Occasional travel to and work at the National Center for Electron Microscopy {http://ncem.lbl.gov/} , Lawrence Berkeley National Laboratory is anticipated to access some of the most advanced TEM instrumentation in the country. Summer research at the Department of Physics, Technical University Chemnitz may also become part of some projects.
The ideal candidate has a diploma of BSc. in crystallography, mineralogy, materials science and engineering, physics, or chemistry, and it interested in both geometrical-structural crystallography and transmission electron microscopy. Computer- and experimental skills are essential for the success of the projects. While some projects will require more experimental skills, some other projects will require more computer skills (notably JScript, Java, C++, Perl, PHP).
Portland State University is with an estimated enrollment of about 30,000 students Oregon’s largest and only urban university. Portland/Oregon {http://www.portlandonline.com/} is the major city in the so called “silicon forest”, i.e. a region where high tech industries blend in with abundant natural beauty, and has consistently been ranked in the top ten of “America’s most livable cities”. Information on student life in Portland {http://www.physics.pdx.edu/gep_files/gep.htm} can be found on our departmental web page. The search will be open until successful candidates have taken up their respective positions.
Applications (CV, references, etc.) should be sent to: Prof. Peter Moeck, Department of Physics, Portland State University, P.O. Box 751, Portland, Oregon 97207-0751
Michael has made an important point the type of DP heater is matched to the original fluid in the DP you cannot just change to another fluid, the fluid has to match the original fluid characteristics, boiling point for instance!
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {michael-at-shaffer.net} To: {protrain-at-emcourses.com} Sent: Tuesday, July 25, 2006 7:40 PM
Hello, I bought a microwave from panasonic with its inverter technology that lowers the wattage of the microwave.
I want to test it to see how well it can do processing for SEM. I was going to try bacteria on a surface, but anyone recommend other sensitive tissues? Leaves from plants maybe?
I'll post the results here if there is interest.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab AOL/IM rakhasha http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
==============================Original Headers============================== 4, 24 -- From gvrdolja-at-nature.berkeley.edu Wed Jul 26 17:44:36 2006 4, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6QMia8U027109 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 17:44:36 -0500 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 3122BC1E33 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 15:44:36 -0700 (PDT) 4, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 4, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 4, 24 -- with ESMTP id 07939-08 for {microscopy-at-microscopy.com} ; 4, 24 -- Wed, 26 Jul 2006 15:44:29 -0700 (PDT) 4, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 4, 24 -- id 66CF3C1E1D; Wed, 26 Jul 2006 15:44:29 -0700 (PDT) 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 2ED48C1E33 4, 24 -- for {microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 15:44:28 -0700 (PDT) 4, 24 -- Date: Wed, 26 Jul 2006 15:44:27 -0700 (PDT) 4, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 4, 24 -- To: microscopy-at-microscopy.com 4, 24 -- Subject: microwave fixation 4, 24 -- Message-ID: {Pine.SOC.4.64.0607261542380.3594-at-nature.Berkeley.EDU} 4, 24 -- MIME-Version: 1.0 4, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 4, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 ==============================End of - Headers==============================
Debby Sherman and I thought it might be of interest for those using ultra-rapid freezing and freeze substitution approaches in their work to get together during the M&M 2006 meeting, in an informal setting. The notion is to share experiences (both good and bad) and ideas in using this methodology, that this will lead to improved success rate in sample preparation. If you are interested, please feel free to join us. The venue is the bar at the Holiday Inn at City Center, where we will have a large table set aside for a congenial setting. The time is Wednesday, August 2, beginning at 5:15 p.m. or so, and continuing until?
With best wishes for a successful M&M 2006,
Howard Berg
Director, Integrated Microscopy Facility Danforth Plant Science Center 975 N. Warson Rd. St. Louis, MO 63132
ph 314-587-1261 fx 314-587-1361 cell 314-378-2409 rhberg-at-danforthcenter.org www.danforthcenter.org visit this educational resource: http://www.danforthcenter.org/Cells/
==============================Original Headers============================== 12, 21 -- From RHBerg-at-danforthcenter.org Wed Jul 26 19:09:22 2006 12, 21 -- Received: from spm1.ddpsc.org (spm1.danforthcenter.org [65.254.111.26]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6R09Mpu006178 12, 21 -- for {Microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 19:09:22 -0500 12, 21 -- Received: from spm1.ddpsc.org (127.0.0.1) by spm1.ddpsc.org (MlfMTA v3.1r24) id hp02340171sm for {Microscopy-at-microscopy.com} ; Wed, 26 Jul 2006 19:09:21 -0500 (envelope-from {RHBerg-at-danforthcenter.org} ) 12, 21 -- Received: from mail02.ddpsc.org ([10.101.0.23]) 12, 21 -- by spm1.ddpsc.org (SonicWALL 4.6.0.7527) 12, 21 -- with ESMTP; Wed, 26 Jul 2006 19:09:21 -0500 12, 21 -- Received: from [10.14.0.20] ([10.14.0.20] unverified) by mail02.ddpsc.org with Microsoft SMTPSVC(5.0.2195.6713); 12, 21 -- Wed, 26 Jul 2006 19:08:32 -0500 12, 21 -- Mime-Version: 1.0 (Apple Message framework v750) 12, 21 -- Content-Transfer-Encoding: 7bit 12, 21 -- Message-Id: {13FDEFCB-A55A-4C64-85D8-07BD373A23D1-at-danforthcenter.org} 12, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 21 -- To: Microscopy-at-microscopy.com 12, 21 -- From: "R. Howard Berg" {rhberg-at-danforthcenter.org} 12, 21 -- Subject: Informal discussion, freeze substitution methodology, at M&M 2006 12, 21 -- Date: Wed, 26 Jul 2006 19:08:29 -0500 12, 21 -- X-Mailer: Apple Mail (2.750) 12, 21 -- X-OriginalArrivalTime: 27 Jul 2006 00:08:32.0040 (UTC) FILETIME=[CB9E9A80:01C6B110] 12, 21 -- X-Mlf-Version: 4.6.0.7527 ==============================End of - Headers==============================
I do believe that the comparatively warm water you are using in the cooling system would also be a factor. 25 degrees is fairly warm. I run my Denton cooling water at 15 degrees.
Ted Dunn The EMscope Co. Ld.
--- protrain-at-emcourses.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi } } Michael has made an important point the type of DP } heater is matched to the } original fluid in the DP you cannot just change to } another fluid, the fluid } has to match the original fluid characteristics, } boiling point for instance! } } Steve Chapman } Senior Consultant Protrain } For electron microscopy consultancy and training } world wide } Tel +44 1280 816512 Fax +44 1280 814007 } Mobile +44 7711 606967 Web www.emcourses.com } } ----- Original Message ----- } X-from: {michael-at-shaffer.net} } To: {protrain-at-emcourses.com} } Sent: Tuesday, July 25, 2006 7:40 PM } Subject: [Microscopy] RE: [Microscopy}RE: Diff. Pump } Problem } } } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } } WCB writes ... } } } } } As suggested by others, Stanton's problem with } his diffusion } } } pump may very well be due to insufficient heat } input from the } } } DP's heater. ... } } } } I remember a similar problem, which could have } well been the heater output } } ... But in this case it was because I had changed } the DP fluid to another } } that had a higher boiling pt. The heater did work } ... But it sure was } } sluggish in the beginning. } } } } Genuinely, Michael Shaffer :o) } } } } SEM/MLA Research Coordinator } } http://www.mun.ca/creait/maf/ } } Inco Innovation Centre } } Memorial University } } St. John's, NL } } } } } } } } ==============================Original } } Headers============================== } } 7, 21 -- From michael-at-shaffer.net Tue Jul 25 } 13:39:13 2006 } } 7, 21 -- Received: from ws6-5.us4.outblaze.com } (ws6-5.us4.outblaze.com } } [205.158.62.152]) } } 7, 21 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } } k6PIdCN0016239 } } 7, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 25 } Jul 2006 13:39:12 -0500 } } 7, 21 -- Received: (qmail 24459 invoked from } network); 25 Jul 2006 } } 18:39:13 -0000 } } 7, 21 -- Received: from unknown (HELO rarewolf) } } (michael-at-shaffer.net-at-205.251.84.119) } } 7, 21 -- by ws6-5.us4.outblaze.com with SMTP; 25 } Jul 2006 18:39:12 -0000 } } 7, 21 -- From: "michael shaffer" } {michael-at-shaffer.net} } } 7, 21 -- To: "MSA Microscopy list" } {Microscopy-at-microscopy.com} } } 7, 21 -- Cc: {bigelow-at-engin.umich.edu} } } 7, 21 -- Subject: RE: [Microscopy] [Microscopy}RE: } Diff. Pump Problem } } 7, 21 -- Date: Tue, 25 Jul 2006 16:09:12 -0230 } } 7, 21 -- Message-ID: } {000b01c6b019$a08639e0$4701a8c0-at-rarewolf} } } 7, 21 -- MIME-Version: 1.0 } } 7, 21 -- Content-Type: text/plain; } } 7, 21 -- charset="us-ascii" } } 7, 21 -- Content-Transfer-Encoding: 7bit } } 7, 21 -- X-Mailer: Microsoft Office Outlook 11 } } 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE } V6.00.2900.2869 } } 7, 21 -- Thread-Index: } AcawBILzsLXAAdW2Qhi/oRSyFMrupwAFIf3g } } 7, 21 -- In-Reply-To: } {200607251608.k6PG81A3025848-at-ns.microscopy.com} } } ==============================End of - } } Headers============================== } } } } } } } ==============================Original } Headers============================== } 7, 33 -- From protrain-at-emcourses.com Wed Jul 26 } 12:37:54 2006 } 7, 33 -- Received: from rd01.mail.x-isp.net } (rd01.mail.x-isp.net [213.253.171.197]) } 7, 33 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k6QHbrAv010973 } 7, 33 -- for {microscopy-at-microscopy.com} ; Wed, 26 } Jul 2006 12:37:53 -0500 } 7, 33 -- Received: from [62.69.65.166] } (helo=smtp-c.mail.legend.co.uk) } 7, 33 -- by rd01.mail.x-isp.net with smtp (Exim } 4.60) } 7, 33 -- (envelope-from {protrain-at-emcourses.com} ) } 7, 33 -- id 1G5nKS-00056m-GH } 7, 33 -- for microscopy-at-microscopy.com; Wed, 26 Jul } 2006 18:37:52 +0100 } 7, 33 -- Received: (qmail 10343 invoked from } network); 26 Jul 2006 17:37:50 -0000 } 7, 33 -- Received: from unknown (HELO advent) } (212.248.149.133) } 7, 33 -- by 0 with SMTP; 26 Jul 2006 17:37:50 } -0000 } 7, 33 -- Message-ID: } {001c01c6b0da$4b29d670$8595f8d4-at-advent} } 7, 33 -- Reply-To: "Steve Chapman" } {protrain-at-emcourses.com} } 7, 33 -- From: "Steve Chapman" } {protrain-at-emcourses.com} } 7, 33 -- To: {michael-at-shaffer.net} } 7, 33 -- Cc: "American Soc" } {microscopy-at-microscopy.com} } 7, 33 -- References: } {200607251840.k6PIeCt1018097-at-ns.microscopy.com} } 7, 33 -- Subject: Re: [Microscopy] RE: } [Microscopy}RE: Diff. Pump Problem } 7, 33 -- Date: Wed, 26 Jul 2006 18:37:59 +0100 } 7, 33 -- Organization: Protrain } 7, 33 -- MIME-Version: 1.0 } 7, 33 -- Content-Type: text/plain; } 7, 33 -- format=flowed; } 7, 33 -- charset="iso-8859-1"; } 7, 33 -- reply-type=original } 7, 33 -- Content-Transfer-Encoding: 7bit } 7, 33 -- X-Priority: 3 } 7, 33 -- X-MSMail-Priority: Normal } 7, 33 -- X-Mailer: Microsoft Outlook Express } 6.00.2900.2527 } 7, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE } V6.00.2900.2527 } 7, 33 -- X-tmReceived: from [62.69.65.166] } (helo=smtp-c.mail.legend.co.uk) } 7, 33 -- by rd01.mail.x-isp.net with smtp id } 1G5nKS-00056m-GH;Wed, 26 Jul 2006 18:37:52 +0100 } ==============================End of - } Headers============================== }
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The matter of changing from one type of diffusion pump fluid to another is a rather complicated one, which is discussed in some detail in Section 5.4.6 (p. 188) of my book 'Vacuum Methods in Electron Microscopy'. (Available from SPI, Ladd, M. E. Taylor, etc.) Theoretically, fluids which have similar slopes for their vapor pressure vs temperature curves, shown in Fig. 5.6 (p. 182), should have similar heats of vaporization, and therefore should require similar heat input from the DP heater, and so should be interchangeable. Note, however, that this plot involves logarithmic scales, so that small differences in slope may involve large differences in actual values. Therefore, just to be on the safe side, I would recommend checking with the pump's manufacturer before making a change. -- Wilbur C. Bigelow, Professor Emeritus Materials Sci. & Engr., Univ. of Michigan Ann Arbor, Michigan 48109-2136 e-mail: bigelow-at-engin.umich.edu; Fx:734-763-4788; Ph:734-975-0858 Address mail to: 2911 Whittier Court Ann Arbor, MI 48104-6731
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Email: dwaugh-at-kent.edu Name: David Waugh
Organization: Kent State University
Title-Subject: [Filtered] SEM mechanical stage parts (Amray 1645)
Question: We have an Amray 1645 with a mechanical stage that has just died. The treaded shaft that actuates the Y axis of the stage has sheared off at the location of the shear pin that holds the gear on the non-threaded part of the shaft. See links for pictures of broken shaft. Does anyone have some spare parts that that they could "part" with? Many Thanks, David
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Question: We are using 6nm-10 beads conjugated to IgG (or IgM) antibodies to detect IgG (or IgM) Monoclonal antibodies bound to a viral surface antigens. Does anyone know the distance(s) from the antigens to the beads?
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Email: camiller-at-anatomy.iupui Name: Caroline Miller
Organization: Indiana University
Title-Subject: [Filtered] FIG on Cryomicroscopy
Question: I would like to let everyone know that a FIG has been approved for Cryomicroscopy. There will be an organizational meeting Tuesday, Aug. 1st from 12:00-12:30PM in Room 308 in Festival Hall during the MM2006 Meeting. I encourage everyone who is interested to attend this meeting. Your input will help to determine the direction of this new FIG. Maybe this is a more formal setting, but still a good way for all us to have a venue for discussing problems and sucesses with high pressure freezing, automatic freeze substitution and cryo ultra microtomy.
Try Fjeld, Inc. A lot of stages were and are still made by them.
gary g.
At 03:10 PM 7/27/2006, you wrote:
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Your question is not so easy to answer. Probably only an approximation can be given. One has to consider at least: * dimensions of antibodies (often described as approximately 10nm max dimension for an IgG) * dimensions of particles (6 or 10 nm in this case) * orientation of antibodies on particle surfaces
If you would consider the first antibody only, sitting on the antigen, and realizing the antibody has a number of hinges in its structure, this would mean that in a worst case assumption assuming full flexibility around the antigen the primary antibody is located within a circle with a radius of about 10nm with the antigen binding site as the geometrical center. The radius will be bigger when working with IgM. Gold conjugates are often imagined as a particle sitting on the Fc- tail of a secondary antibody. Some secondaries may actually be oriented that way, but it is very likely that some secondaries are oriented side-on on the gold particle surface. So there may very well be a range of conjugate sizes if you like. Again, a worst case assumption would probably bring the center of a 10nm particle of a secondary conjugate about 15 nm away (for an IgG conjugate) from the binding site on the primary.
Add all this up and the worst case assumption amounts to an area of probability with a radius of 25nm within which the centre of the gold particle should be located. That is what the situation may be as long as the specimen is in a buffer. During drying the dimensions of the whole structure may change while collapsing onto the specimen. Is it reaonable to assume worst case conditions? From our own work in the past, even three step labeling involving an unlabelled secondary antibody and protein A gold conjugate to label the outer segment of a membrane protein in bacteria still showed the majority of the particles located outside the cells and not projected over the periplasmic space or even the outer membrane.
What has often made me wonder in working with viruses or Virus Like Particles (VLP) and immunolabelling is that the gold label is often surrounding the virus particles rather than actually covering the structures.
In practice, and theoretically for resolution reasons, it may be worthwhile to reduce the labelling to a one step one, with the gold particle sitting directly on the primary antibody, or even on an Fab fragment of the primary. One step labelling of course also has its disadvantage in that the presence of the gold particle on the primary antibody may result in steric hindrance. Using an ultra small colloidal particle or covalently coupled particle and post-labelling enhancement would be the best choice to reduce those risks.
Well, I am not sure if all this helps, please feel free to get in touch if you would like to discuss this further.
Cheers
Jan Leunissen
Aurion - President Electron Microscopy Research Advisor Costerweg 5 Dept Anatomy and Structural Biology 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4795465 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz
David Waugh wrote: ========================================================== Question: We have an Amray 1645 with a mechanical stage that has just died. The treaded shaft that actuates the Y axis of the stage has sheared off at the location of the shear pin that holds the gear on the non-threaded part of the shaft. See links for pictures of broken shaft. Does anyone have some spare parts that that they could "part" with? Many Thanks, David =============================================== Contact
Mr. Clark Houghton Secondary Images 2371 Emery Lane Winchester, OH 45697
They are a "third party" service firm and are almost a neighbor of yours. They specialize in servicing AMRAY SEMs. I think they could help you with this problem.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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Email: susan.vanhorn-at-sunysb.edu Name: Susan Van Horn
Organization: SUNY -at- StonyBrook
Title-Subject: [Filtered] glow discharge
Question: I am getting mixed information concerning glow discharged grids: once they are glow discharged how long does it last??? and how is it accomplished?? thanks sue
On Jul 28, 2006, at 2:15 PM, susan.vanhorn-at-sunysb.edu wrote:
} Question: I am getting mixed information concerning glow discharged } grids: once they are glow discharged how long does it last??? and how } is it accomplished?? } Dear Sue, The effect of glow-discharging--to make the carbon hydrophyllic--fades over time, and how long it lasts depends on just how hydrophyllic the carbon must be to satisfy your particular need. For cryoEM, we have found that putting the material on the grid for freezing is best done within a few minutes after a 2 minute glow-discharge step. However, for other specimens a longer interval is OK, and for bacteriorhodopsin, the carbon surface should be "aged" over a weekend for best results. An additional complication is that the conversion of the surface from hydrophyllic to hydrophobic depends on the environmental conditions, so it can vary from lab to lab. It is accomplished by the bombardment of the carbon by oxygen or nitrogen ions (or whatever else is in the plasma); in order to make extremely hydrophyllic grids, amyl amine has been introduced into the glow-discharge volume, so in that case I think that molecular ions or fragments from the amyl amine must be particularly effective, but I don't know which component(s) is the actual active one. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Fri Jul 28 16:47:42 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6SLlgX2020748 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 28 Jul 2006 16:47:42 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP 4, 22 -- id C0B4C3545D; Fri, 28 Jul 2006 14:47:41 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP 4, 22 -- id 255C92EF86; Fri, 28 Jul 2006 14:47:36 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200607282115.k6SLFq72000454-at-ns.microscopy.com} 4, 22 -- References: {200607282115.k6SLFq72000454-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {285b9ef1b3a605900886a7a66441976c-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: glow discharge 4, 22 -- Date: Fri, 28 Jul 2006 14:49:15 -0700 4, 22 -- To: susan.vanhorn-at-sunysb.edu, microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
I am working on a MSc project in which I am studying the effect of microwave exposure to water during freezing, in the context of electron microscopy sample preparation.
A couple of papers published indicate a reduction in the size of ice crystals when water (including that in biological samples) is frozen in the presence of microwaves (references below).
There is even a commercially available instrument, the Eiko MF-2 (see Eiko Instruments' website at http://www.kagaku.com/eiko/e-products/ electron/mf_2.html), which that company claims provides a reduction in ice crystal size when freezing (I have no commercial interest whatsoever in this company, by the way).
I would be very pleased to hear from anyone who has studied this effect (even if the work was unpublished) or knows of any other work on the subject. It would also be interesting to hear from anyone who has actually used the MF-2!
Best regards
Richard
References: Hanyu, Y., Ichikawa, M. and Matsumoto, G: An improved cryofixation method: cryoquenching of small tissue blocks during microwave irradiation. Journal of Microscopy, 165:Pt 2 255-271. February 1992
Jackson, T.H., Ungan, U., Critser, J.K. and Gao, D: Novel microwave technology for cryopreservation of biomaterials by suppression of apparent ice formation. Cryobiology 34, 363-372. 1997
Richard Easingwood Otago Centre for Electron Microscopy c/- Department of Anatomy & Structural Biology University of Otago, Dunedin, New Zealand ph: 0064 3 479 7301 cell: 021 222 4759 fax: 0064 3 479 7254 or 479 5086 http://ocem.otago.ac.nz
==============================Original Headers============================== 14, 20 -- From richard.easingwood-at-stonebow.otago.ac.nz Sat Jul 29 17:23:30 2006 14, 20 -- Received: from mailhub1.otago.ac.nz (mailhub1.otago.ac.nz [139.80.64.218]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k6TMNTPS003051 14, 20 -- for {Microscopy-at-microscopy.com} ; Sat, 29 Jul 2006 17:23:29 -0500 14, 20 -- Received: from galadriel.otago.ac.nz (galadriel.otago.ac.nz [139.80.64.213]) 14, 20 -- by mailhub1.otago.ac.nz (8.13.6/8.13.6) with ESMTP id k6TMNSVs019262 14, 20 -- for {Microscopy-at-microscopy.com} ; Sun, 30 Jul 2006 10:23:28 +1200 14, 20 -- Received: from dialin093096.otago.ac.nz ([139.80.93.96]) 14, 20 -- by galadriel.otago.ac.nz with esmtp (Exim 4.50) 14, 20 -- id 1G6xDT-0001cs-7x 14, 20 -- for Microscopy-at-microscopy.com; Sun, 30 Jul 2006 10:23:27 +1200 14, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 14, 20 -- Content-Transfer-Encoding: 7bit 14, 20 -- Message-Id: {27F38189-5C17-48CD-9925-D5BEE529E611-at-stonebow.otago.ac.nz} 14, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 14, 20 -- To: Microscopy-at-microscopy.com 14, 20 -- From: Richard Easingwood {richard.easingwood-at-stonebow.otago.ac.nz} 14, 20 -- Subject: Microwave assisted freezing 14, 20 -- Date: Sun, 30 Jul 2006 10:23:25 +1200 14, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
Susan Van Horn wrote: ==================================================== Question: I am getting mixed information concerning glow discharged=20 grids: once they are glow discharged how long does it last??? and how is it accomplished?? thanks ====================================================== We have found two different approaches can "work":
a) If the glow discharge is done in a small table top plasma etcher, such as the SPI Plasma Prep II, see URL http://www.2spi.com/catalog/instruments/etchers1.shtml, using either an "air" or nitrogen plasma, carbon coated grids can be made hydrophilic and they will maintain their hydrophilic nature for 30-60 days before they start to lose these enhanced properties. The "mixed information" you reference could be due to the fact that there are a number of different systems that produce a "glow discharge". The SPI system operates at 100 watts. Systems of lower power seem to produce grids that will retain the enhanced properties for a shorter period of time.
b) Evaporation of a molecular layer of Victawet, see URL http://www.2spi.com/catalog/spec_prep/evapor_3c.shtml This is a phosphate based water soluble surfactant so it could possibly interfere with at least some experiments. But if you can get away with using it, it will leave the grids with their enhanced properties essentially "forever".
Disclaimer: SPI Supplies offers both the SPI Plasma Prep II Plasma Etcher and also, Victawet, so we have a vested interest in seeing more people using both products.
Chuck ================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President=20 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
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I don't know the best stain for your polymer. However, I know that osmium tetroxide will stain any unsaturation in your material and, very importantly, will probably stain little else in the molecule. Specifically, OsO4 adds across the C=C bond. By comparison, ruthenium has much less specificity since it will stain the C=C bond and other hydrocarbon components of the molecule.
Staining can, and probably should, be done in the vapor phase. Affix your specimen to the inside of the cap to a vial containing OsO4 solution and stain overnight. Degas for several hours. Remember that OsO4 is toxic and all work should be done in a good fume hood.
Regards,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
kmoulton-at-usm.m aine.edu To gary.m.brown-at-exxonmobil.com 07/28/06 04:19 cc PM Subject [Microscopy] viaWWW: TEM staining Please respond methods to kmoulton-at-usm.m aine.edu
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Samsung Austin Semiconductor is currently looking for a TEM Engineer to join its Materials & Analysis Team. The M&A team is responsible for surface analysis and fab inspection. The surface analysis team will work primarily with SIMS, Auger analysis and FIB and TEM imaging. The fab inspection team will focus primarily on bevel and edge defects and tool level defect fingerprinting.
Job Duties
* Manage TEM Microscope * Perform TEM imaging and summary write ups * Oversee TEM Sample prep and provide feedback * Work with unit process and product technology engineers to resolve device and in line issues
Requirements
* BS in EE, ChemE, or Material Science * TEM microscope operation * DRAM device knowledge * FLASH device knowledge
Please send resumes or questions to Alison Rao at arao-at-sas.samsung.com
"Samsung Austin Semiconductor" made the following annotations. ------------------------------------------------------------------------------ NOTICE : This email message is for the sole use of the intended recipient(s) and may contain confidential and /or privileged information. Any unauthorized review, use, disclosure or distribution is prohibited. If you are not the intended recipient, please contact the sender by reply email and destroy all copies, including without limitation electronic or printed versions, of this message and any attachments . ==============================================================================
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At 13:16 25/07/2006 -0500, you wrote: Hi Dorota, I have a few things to suggest, possibly not required but this is not obvious from the information you provide.
Although the blocks seem hard the fact that they are sticky indicates that there is a polymerisation issue.
I would firstly check out the catalyst. It is supplied by many manufacturers in a very much different condition than previously and would probably be improved if you were to routinely dry it just prior to use (30mins in a c. 45'C oven). You may also be using a 'bad batch of catalyst. We routinely and regularly make our own GMA (JB4 equivalent) and had to return a recent batch of benzoyle peroxide (catalyst) to Sigma as polymerisation was drastically altered following a recent change in production method (despite assurances that it was a direct replacement for the previous chemical).
Your accelerator may have been contaminated or deteriorated during a lengthy period following initial opening of the bottle. We buy the minimum volume available, and replace all our accelerators on a 6 monthly basis - they definitely deteriotrate on storage and the purchase cost is negligible.
Polymerisation - you don't mention restricting air contact. Despite using vacuum, air contact should be minimised by capping or sealing the molds, or by polymerisation in an inert gas atmosphere (such as nitrogen).
Hope this helps, Alastair
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Alastair McKinnon Histology & EM Facility Manager University of Aberdeen, Institute of Medical Science Foresterhill, Aberdeen, AB25 2ZD tel: +44(0) 1224 552923 (office); +44(0) 1224 555911 (lab) fax: +44(0) 1224 559533; email: a.d.mckinnon-at-abdn.ac.uk www.abdn.ac.uk/ims/h-em
==============================Original Headers============================== 12, 18 -- From a.d.mckinnon-at-abdn.ac.uk Tue Aug 1 03:32:50 2006 12, 18 -- Received: from mailhub3.abdn.ac.uk (mailhub3.abdn.ac.uk [139.133.7.13]) 12, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k718Wosx020502 12, 18 -- for {Microscopy-at-Microscopy.Com} ; Tue, 1 Aug 2006 03:32:50 -0500 12, 18 -- Received: from med-0069.ims.abdn.ac.uk ([139.133.159.90] helo=med-0069.abdn.ac.uk) 12, 18 -- by mailhub3.abdn.ac.uk with esmtp (Exim 4.52) 12, 18 -- id 1G7pgF-0006Ur-0k; Tue, 01 Aug 2006 09:32:47 +0100 12, 18 -- Message-Id: {5.2.1.1.0.20060801090513.01118408-at-mailms.abdn.ac.uk} 12, 18 -- X-Sender: pat081-at-mailms.abdn.ac.uk 12, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 12, 18 -- Date: Tue, 01 Aug 2006 09:32:44 +0100 12, 18 -- To: wadowska-at-upei.ca 12, 18 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 12, 18 -- Subject: Re: [Microscopy] JB-4 resin problems 12, 18 -- Cc: Microscopy-at-Microscopy.Com 12, 18 -- In-Reply-To: {200607251816.k6PIGI2o014502-at-ns.microscopy.com} 12, 18 -- Mime-Version: 1.0 12, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
The Faculty of Mathematics and Natural Sciences of the University of Groningen invites applications for a tenure track assistant professorship in Surface Science, with specialisation in scanning tunneling microscopy - for details about the position, requirements and application procedure see http://www.rug.nl/fwn/vacatures/vacaturesfwn/206151.
I would most grateful if you could kindly alert suitable candidates to this oppurtunity.Deadline is September 15th, 2006.
Please kindly note that this kind of position foresees the promotion to associate and further on to full professor if certain criteria are fullfilled.
Thank you very much in advance for your help.
Sincerely yours, Petra Rudolf
Prof. Dr. Petra Rudolf Materials Science Centre University of Groningen Nijenborgh 4 9747 AG Groningen, the Netherlands phone: +(31)50-363 4736 e-mail: P.Rudolf-at-rug.nl web page: http://www.surfacesthinfilms.fmns.rug.nl/
I have been curious for some time now about how the quality/age of a certain batch of G-1 (or its equivalents) affects cross-sectioning properties for TEM samples. The shelf life listed on the small bottles purchased from Gatan indicate 1 year, as well as the larger bottles from epo-tek. Has anyone been able to relate poor adhesion, poor ion-milling, etc. to the age or quality of the epoxy? I know people who have used the same bottles of epo-tek for years with no problems.
Thanks, Leslie
Leslie Krupp (Thompson) IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 4, 29 -- From lkrupp-at-us.ibm.com Tue Aug 1 16:05:11 2006 4, 29 -- Received: from e4.ny.us.ibm.com (e4.ny.us.ibm.com [32.97.182.144]) 4, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k71L5Avb024845 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 16:05:10 -0500 4, 29 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 4, 29 -- by e4.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k71L56XP011677 4, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=FAIL) 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:07 -0400 4, 29 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) 4, 29 -- by d01relay02.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k71L56cA168446 4, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 4, 29 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) 4, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k71L56op020767 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 4, 29 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 4, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k71L56SH020745 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 4, 29 -- To: Microscopy-at-microscopy.com 4, 29 -- Subject: quality/shelf life of G-1 epoxy, epo-tek, etc. 4, 29 -- MIME-Version: 1.0 4, 29 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 4, 29 -- Message-ID: {OF9A56A70D.BC20401E-ON852571BD.00702791-882571BD.0073D168-at-us.ibm.com} 4, 29 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 4, 29 -- Date: Tue, 1 Aug 2006 14:05:03 -0700 4, 29 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF269 | June 22, 2006) at 4, 29 -- 08/01/2006 17:05:06, 4, 29 -- Serialize complete at 08/01/2006 17:05:06 4, 29 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
I have used batches of Epo-Tek 353ND that I purchased for longer than two years. I started to increase the amount of harder (as measured by drops) when it got older. Actually, what I did was decrease the number of resin drops and kept the hardner drops to 1.
Finally I mixed what was left and cured it on a hot plate. Warning, if you do this, use a big enough container to catch the boil out. I destroyed the surface of a good hot plate.
If you store it at room temp and not in direct sunlight, I think that you can go about 2 years. After that replace it. I did not see a fall off in the adhesion capability of the epoxy as it aged.
In a past life, I've used the G1, with the same results. Since I believe that it is the same stuff as the 353-ND, I will not say any more about it.
SBT does not sell the Epo-Tek 353-ND, we refer our customers to Epoxy Technology, Inc whose website is www.epotek.com.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
-------- Original Message -------- } From: lkrupp-at-us.ibm.com } Sent: Tuesday, August 01, 2006 5:09 PM } To: Walck-at-SouthBayTech.com } Subject: [Microscopy] quality/shelf life of G-1 epoxy, epo-tek, etc. } } ---------------------------------------------------------------------------- } 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 listers- } } I have been curious for some time now about how the quality/age of a } certain batch of G-1 (or its equivalents) affects cross-sectioning } properties for TEM samples. The shelf life listed on the small bottles } purchased from Gatan indicate 1 year, as well as the larger bottles from } epo-tek. Has anyone been able to relate poor adhesion, poor ion-milling, } etc. to the age or quality of the epoxy? I know people who have used the } same bottles of epo-tek for years with no problems. } } Thanks, } Leslie } } Leslie Krupp (Thompson) } IBM Almaden Research } 650 Harry Road, K19/D1 } San Jose, CA 95120-6099 } (408) 927-3856 } } ==============================Original Headers============================== } 4, 29 -- From lkrupp-at-us.ibm.com Tue Aug 1 16:05:11 2006 } 4, 29 -- Received: from e4.ny.us.ibm.com (e4.ny.us.ibm.com [32.97.182.144]) } 4, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k71L5Avb024845 } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 16:05:10 -0500 } 4, 29 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) } 4, 29 -- by e4.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k71L56XP011677 } 4, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=FAIL) } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:07 -0400 } 4, 29 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) } 4, 29 -- by d01relay02.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k71L56cA168446 } 4, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 } 4, 29 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) } 4, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k71L56op020767 } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 } 4, 29 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) } 4, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k71L56SH020745 } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 } 4, 29 -- To: Microscopy-at-microscopy.com } 4, 29 -- Subject: quality/shelf life of G-1 epoxy, epo-tek, etc. } 4, 29 -- MIME-Version: 1.0 } 4, 29 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 } 4, 29 -- Message-ID: {OF9A56A70D.BC20401E-ON852571BD.00702791-882571BD.0073D168-at-us.ibm.com} } 4, 29 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} } 4, 29 -- Date: Tue, 1 Aug 2006 14:05:03 -0700 } 4, 29 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF269 | June 22, 2006) at } 4, 29 -- 08/01/2006 17:05:06, } 4, 29 -- Serialize complete at 08/01/2006 17:05:06 } 4, 29 -- Content-Type: text/plain; charset="US-ASCII" } ==============================End of - Headers==============================
==============================Original Headers============================== 5, 20 -- From walck-at-southbaytech.com Wed Aug 2 00:24:03 2006 5, 20 -- Received: from smtp14.safesecureweb.com (smtp14.safesecureweb.com [66.241.211.108]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k725O26x011759 5, 20 -- for {microscopy-at-microscopy.com} ; Wed, 2 Aug 2006 00:24:02 -0500 5, 20 -- Received: from mail15.safesecureweb.com (ironhide.ad.safesecureweb.com [192.168.2.180]) 5, 20 -- by smtp14.safesecureweb.com (Spam Firewall) with ESMTP 5, 20 -- id B2B6F15BBCF2; Wed, 2 Aug 2006 01:24:01 -0400 (EDT) 5, 20 -- MIME-Version: 1.0 5, 20 -- Date: Wed, 2 Aug 2006 01:23:40 -0400 5, 20 -- Content-Type: text/plain; 5, 20 -- charset=iso-8859-1 5, 20 -- Subject: re: [Microscopy] quality/shelf life of G-1 epoxy, epo-tek, etc. 5, 20 -- From: Scott Walck {walck-at-southbaytech.com} 5, 20 -- Reply-To: Walck-at-southbaytech.com 5, 20 -- To: {lkrupp-at-us.ibm.com} , {microscopy-at-microscopy.com} 5, 20 -- CC: 5, 20 -- Message-ID: {5b68b38b80844900b11aa30fb3f195ea-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 k725O26x011759 ==============================End of - Headers==============================
I'm new to this group and the industry and am still on a learning curve. I want to learn about the field and need a good place to start. With that in mind, I'm hoping someone can refer me to some good sources of information in the field of confocal and in particular, imaging spectral microscopy.
X-from the technical side, I would like to get some overviews of the technology, the applications the suppliers and the pros and the cons. From the business side I'm looking for any reports on market sizes and where can I find general pricing information to get a sense of what things cost? Thanks in advance! I'm interested in any sources of information (i.e. people, websites, specific magazine issues and other literature).
Thanks in advance!
Alexandre Fong, Optronic Laboratories, Inc.
==============================Original Headers============================== 7, 25 -- From aFong-at-olinet.com Thu Aug 3 12:14:52 2006 7, 25 -- Received: from orlandotelco.net (sun-msg-1.orlandotelco.net [216.54.161.51]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k73HEnnF023720 7, 25 -- for {Microscopy-at-Microscopy.Com} ; Thu, 3 Aug 2006 12:14:51 -0500 7, 25 -- Received: from VPSalesLT (unverified [64.132.123.34]) 7, 25 -- by orlandotelco.net (SurgeMail 3.7b6) with ESMTP id 13834118 7, 25 -- for {Microscopy-at-Microscopy.Com} ; Thu, 03 Aug 2006 12:40:58 -0400 7, 25 -- Reply-To: {afong-at-olinet.com} 7, 25 -- From: "Alex Fong" {aFong-at-olinet.com} 7, 25 -- To: {Microscopy-at-Microscopy.Com} 7, 25 -- Subject: LM Need technical and business resources on hyper/multi-spectral imaging microscopy 7, 25 -- Date: Thu, 3 Aug 2006 13:14:49 -0400 7, 25 -- Organization: Optronic Laboratories 7, 25 -- Message-ID: {007801c6b720$536f5700$0c01a8c0-at-VPSalesLT} 7, 25 -- MIME-Version: 1.0 7, 25 -- Content-Type: text/plain; 7, 25 -- charset="us-ascii" 7, 25 -- X-Priority: 3 (Normal) 7, 25 -- X-MSMail-Priority: Normal 7, 25 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 7, 25 -- Importance: Normal 7, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 7, 25 -- X-Server: High Performance Mail Server - http://surgemail.com r=809480741 7, 25 -- Content-Transfer-Encoding: 8bit 7, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k73HEnnF023720 ==============================End of - Headers==============================
The Centre for Microscopy and Microanalysis at the University of Queensland (Brisbane, Australia) is seeking a Scientific Officer to work in the Cryo Electron Microscopy facility. The facility is well equipped with a Tecnai F30 (300kV, FEG, GIF), a Tecnai 12 (120kV, LaB6) and a JEOL 1011 TEM, and also a range of ancillary sample preparation equipment (High Pressure Freezers, Vitrobot, etc). The research performed here includes room temperature and cryo tomography, single particle analysis, energy filtered imaging of biological and some materials samples and routine biological TEM imaging.
The full job description can be obtained online at http://www.seek.com.au/users/apply/index.ascx?Sequence=10&PageNumber=1&JobID =7358227&msid=1&Keywords=
The closing date for applications is the 25th of August, 2006. Any questions regarding the position should be directed to me by email (j.riches-at-uq.edu.au).
Regards, Jamie
Jamie Riches Scientific/Technical Manager Centre for Microscopy and Microanalysis University of Queensland Ph: +61 7 3346 2935 Fax: +61 7 3346 2101 Email: j.riches-at-uq.edu.au
==============================Original Headers============================== 14, 22 -- From j.riches-at-uq.edu.au Fri Aug 4 00:04:57 2006 14, 22 -- Received: from imb.uq.edu.au (mail.imb.uq.edu.au [130.102.113.156]) 14, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7454sge013604 14, 22 -- for {microscopy-at-microscopy.com} ; Fri, 4 Aug 2006 00:04:57 -0500 14, 22 -- X-ExtScanner: Niversoft's FindAttachments (free) 14, 22 -- Received: from [130.102.116.81] (HELO AWMJAMIER) 14, 22 -- by imb.uq.edu.au (CommuniGate Pro SMTP 5.0.6) 14, 22 -- with ESMTP id 10616841; Fri, 04 Aug 2006 15:04:52 +1000 14, 22 -- Reply-To: {j.riches-at-uq.edu.au} 14, 22 -- From: "Jamie Riches" {j.riches-at-uq.edu.au} 14, 22 -- To: {microscopy-at-microscopy.com} 14, 22 -- Cc: "'Jill Lumsdale'" {j.lumsdale-at-uq.edu.au} 14, 22 -- Subject: Job Opening: Scientific Officer at Uni. of Queensland, Australia 14, 22 -- Date: Fri, 4 Aug 2006 15:00:02 +1000 14, 22 -- Message-ID: {00f501c6b782$d8201310$51746682-at-AWMJAMIER} 14, 22 -- MIME-Version: 1.0 14, 22 -- Content-Type: text/plain; 14, 22 -- charset="us-ascii" 14, 22 -- Content-Transfer-Encoding: 7bit 14, 22 -- X-Mailer: Microsoft Office Outlook 11 14, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 14, 22 -- Thread-Index: Aca3gtf+DOLmfForS3m+MggQiGZosw== ==============================End of - Headers==============================
If anybody out there is using an HRI delay unit with the Picostar image intensifier, we would very much like to speak with you to compare technical details and to seek assistance on troubleshooting.
We'd be happy to communicate by email or to set up a time to call you.
Thank you!! ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
==============================Original Headers============================== 4, 22 -- From cammer-at-aecom.yu.edu Fri Aug 4 08:11:03 2006 4, 22 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k74DB2ul030854 4, 22 -- for {microscopy-at-microscopy.com} ; Fri, 4 Aug 2006 08:11:03 -0500 4, 22 -- Received: from mailvx.aecom.yu.edu (mailvx.aecom.yu.edu [129.98.1.17]) 4, 22 -- by mailgw.aecom.yu.edu (8.12.11.20060308/8.12.11) with SMTP id k74DArGZ014121 4, 22 -- for {microscopy-at-microscopy.com} ; Fri, 4 Aug 2006 09:11:02 -0400 4, 22 -- Received: from post.aecom.yu.edu ([129.98.1.100]) 4, 22 -- by mailvx.aecom.yu.edu (SAVSMTP 3.1.1.32) with SMTP id M2006080409110202270 4, 22 -- for {microscopy-at-microscopy.com} ; Fri, 04 Aug 2006 09:11:02 -0400 4, 22 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.30.137]) 4, 22 -- by post.aecom.yu.edu (Postfix) with ESMTP id 77A8D25 4, 22 -- for {microscopy-at-microscopy.com} ; Fri, 4 Aug 2006 09:11:02 -0400 (EDT) 4, 22 -- Message-Id: {5.2.1.1.2.20060804091011.011f1a28-at-mailserver.aecom.yu.edu} 4, 22 -- X-Sender: cammer-at-mailserver.aecom.yu.edu 4, 22 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 4, 22 -- Date: Fri, 04 Aug 2006 09:13:06 -0400 4, 22 -- To: microscopy-at-microscopy.com 4, 22 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 4, 22 -- Subject: HRI Picostar, Lavision software question 4, 22 -- Mime-Version: 1.0 4, 22 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both javaidqazi-at-kemet.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: javaidqazi-at-kemet.com Name: javaid qazi
Organization: Kemet Electronics
Title-Subject: [Filtered] 3D stereo microscopy, Height/Roughness profile
Question: I am looking for a commercial lab which can do height profiles by optical/digital stereo microscopy on samples. Basically can give the depth/height of different features on the surface of a part. I know Keyence VHX series can do it, certainly there are other manufacturers as well. Just looking for a place where I can get this done. Preferably on east coast. Thanks a lot.
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 fisher.phyllis-at-mayo.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: Re: Gordon Ante Vrodoljak / microwave fixation tests I am interested in your results with regard to fixation on isolated animal cells without the usual chemical fixatives.
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 smythen-at-musc.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: smythen-at-musc.edu Name: Nancy Smythe
Organization: Medical University of South Carolina
Title-Subject: [Filtered] Processing tissue for light microscopy
Question: I need to process and section whole neonate mice for pariffin. Anyone have a protocol? I am also open for suggestions. I will be fixing over the weekend.
Thanks for your help.
Nancy
Nancy Smythe Department of Otolaryngology Head and Neck Surgery Medical University of South Carolina 843-792-8835 843-792-0368 Fax
Dear SEM techs: My 6100 has an intermittent in which the on-screen characters freeze up, the coarse probe current will not respond, but the focus and magnification controls still operate normally. Pressing the reset button only makes the CRT flicker once but nothing resets--i.e. the magnification and focus don't change, and the on-screen characters stay frozen. Typically, after powering the console off and on, the CRT is covered in random characters, and no image can be formed (i.e. the magnification and focus controls are unresponsive). The problem can disappear for hours at a time, or it can occur almost constantly.
We have tried all manner of reseating PCBs, reconnecting connectors, using freeze spray on the ICs, etc. Nothing affects it in any consistent way.
The Jeol schematics are similar to the actual circuits, but when examined more closely, actual the display circuit boards, for example, are quite different from the schematics, with more than double the number of ICs actually present than shown in the drawings.
Questions: 1. Is there a systematic methodology for isolating the trouble? 2. The local Jeol service office has advised that replacement PCBs are unavailable due to the age of the scope. Are there dealers in used SEM parts? I.e. SEM parts recyclers? 3. Does anyone have a service manual? 4. Are accurate schematics available? 5. Is there such a thing as a replacement console with modern electronics? 6. Is there a list server just for SEM techs?
I have nicely organized and annotated pdf versions of the schematics and the operator's manual that I can offer as a token of my appreciation for any help.
Thanks! Bernard Cuzzillo Berkeley, CA USA
==============================Original Headers============================== 6, 14 -- From bernard-at-bearinc.com Sun Aug 6 12:46:24 2006 6, 14 -- Received: from bearinc.com (bearinc.com [64.62.217.44]) 6, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k76HkNLq022494 6, 14 -- for {Microscopy-at-microscopy.com} ; Sun, 6 Aug 2006 12:46:24 -0500 6, 14 -- Received: from 24.7.101.58 ([24.7.101.58]) by bearinc.com for {Microscopy-at-microscopy.com} ; Sun, 6 Aug 2006 10:46:22 -0700 6, 14 -- Message-ID: {44D62AED.8000406-at-bearinc.com} 6, 14 -- Date: Sun, 06 Aug 2006 10:46:21 -0700 6, 14 -- From: Bernard Cuzzillo {bernard-at-bearinc.com} 6, 14 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 6, 14 -- MIME-Version: 1.0 6, 14 -- To: Microscopy-at-microscopy.com 6, 14 -- Subject: Jeol JSM-6100 electronics troubleshooting 6, 14 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 14 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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I obviously have a vested interest but why don't you contact JEOL USA?
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==============================Original Headers============================== 4, 17 -- From larry-at-cymru.freewire.co.uk Sun Aug 6 14:22:20 2006 4, 17 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k76JMJPO001632 4, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 6 Aug 2006 14:22:20 -0500 4, 17 -- Received: from [217.154.254.152] ([217.154.254.152]) 4, 17 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k76JMA4s021518; 4, 17 -- Sun, 6 Aug 2006 20:22:13 +0100 4, 17 -- Mime-Version: 1.0 4, 17 -- Message-Id: {p06210200c0fbf14d59bc-at-[217.154.252.73]} 4, 17 -- In-Reply-To: {200608061748.k76Hmr1I024797-at-ns.microscopy.com} 4, 17 -- References: {200608061748.k76Hmr1I024797-at-ns.microscopy.com} 4, 17 -- Date: Sun, 6 Aug 2006 20:21:15 +0100 4, 17 -- To: bernard-at-bearinc.com 4, 17 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 4, 17 -- Subject: Re: [Microscopy] Jeol JSM-6100 electronics troubleshooting 4, 17 -- Cc: Microscopy-at-MSA.Microscopy.Com 4, 17 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
If you run out of options try these guys. They do component level repair on almost everything. I had a problem with my Kevex monitor and they fixed it for a flat fee. They also fix all the old crap for the FAA. I have no association with them. I am setting up a JEOL 6300F and am looking for a manual if you come across any.
Thanks!
Tom Kaye
SMH Electronics West Wareham MA 508-291-7447
-----Original Message----- X-from: bernard-at-bearinc.com [mailto:bernard-at-bearinc.com] Sent: Sunday, August 06, 2006 12:54 PM To: tom-at-tomkaye.com
Dear SEM techs: My 6100 has an intermittent in which the on-screen characters freeze up, the coarse probe current will not respond, but the focus and magnification controls still operate normally. Pressing the reset button only makes the CRT flicker once but nothing resets--i.e. the magnification and focus don't change, and the on-screen characters stay frozen. Typically, after powering the console off and on, the CRT is covered in random characters, and no image can be formed (i.e. the magnification and focus controls are unresponsive). The problem can disappear for hours at a time, or it can occur almost constantly.
We have tried all manner of reseating PCBs, reconnecting connectors, using freeze spray on the ICs, etc. Nothing affects it in any consistent way.
The Jeol schematics are similar to the actual circuits, but when examined more closely, actual the display circuit boards, for example, are quite different from the schematics, with more than double the number of ICs actually present than shown in the drawings.
Questions: 1. Is there a systematic methodology for isolating the trouble? 2. The local Jeol service office has advised that replacement PCBs are unavailable due to the age of the scope. Are there dealers in used SEM parts? I.e. SEM parts recyclers? 3. Does anyone have a service manual? 4. Are accurate schematics available? 5. Is there such a thing as a replacement console with modern electronics? 6. Is there a list server just for SEM techs?
I have nicely organized and annotated pdf versions of the schematics and the operator's manual that I can offer as a token of my appreciation for any help.
Thanks! Bernard Cuzzillo Berkeley, CA USA
==============================Original Headers============================== 6, 14 -- From bernard-at-bearinc.com Sun Aug 6 12:46:24 2006 6, 14 -- Received: from bearinc.com (bearinc.com [64.62.217.44]) 6, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k76HkNLq022494 6, 14 -- for {Microscopy-at-microscopy.com} ; Sun, 6 Aug 2006 12:46:24 -0500 6, 14 -- Received: from 24.7.101.58 ([24.7.101.58]) by bearinc.com for {Microscopy-at-microscopy.com} ; Sun, 6 Aug 2006 10:46:22 -0700 6, 14 -- Message-ID: {44D62AED.8000406-at-bearinc.com} 6, 14 -- Date: Sun, 06 Aug 2006 10:46:21 -0700 6, 14 -- From: Bernard Cuzzillo {bernard-at-bearinc.com} 6, 14 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 6, 14 -- MIME-Version: 1.0 6, 14 -- To: Microscopy-at-microscopy.com 6, 14 -- Subject: Jeol JSM-6100 electronics troubleshooting 6, 14 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 14 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
==============================Original Headers============================== 17, 21 -- From tom-at-tomkaye.com Sun Aug 6 19:09:22 2006 17, 21 -- Received: from tomkaye.com (mxgateway.techpro.com [64.4.207.8]) 17, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7709MbT015147 17, 21 -- for {Microscopy-at-microscopy.com} ; Sun, 6 Aug 2006 19:09:22 -0500 17, 21 -- Received: from TKLAPTOP [70.213.198.83] by tomkaye.com with ESMTP 17, 21 -- (SMTPD32-8.14) id A4ADA410140; Sun, 06 Aug 2006 19:09:17 -0500 17, 21 -- From: "Tom" {tom-at-tomkaye.com} 17, 21 -- To: {Microscopy-at-microscopy.com} 17, 21 -- Subject: RE: [Microscopy] Jeol JSM-6100 electronics troubleshooting 17, 21 -- Date: Sun, 6 Aug 2006 19:09:19 -0500 17, 21 -- Message-ID: {LHECKJAKKIOGKDBHHJMLOEODCKAA.tom-at-tomkaye.com} 17, 21 -- MIME-Version: 1.0 17, 21 -- Content-Type: text/plain; 17, 21 -- charset="iso-8859-1" 17, 21 -- Content-Transfer-Encoding: 7bit 17, 21 -- X-Priority: 3 (Normal) 17, 21 -- X-MSMail-Priority: Normal 17, 21 -- X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2911.0) 17, 21 -- In-reply-to: {200608061753.k76Hro4V030285-at-ns.microscopy.com} 17, 21 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1165 17, 21 -- Importance: Normal ==============================End of - Headers==============================
All, Sorry about that, I guess the way I listed it you could interpret it both ways. Yes that's the company and I don't have any involvement with SMH other than being a customer.
Tom
-----Original Message----- X-from: Ritchie Sims [mailto:r.sims-at-auckland.ac.nz] Sent: Sunday, August 06, 2006 8:28 PM To: tom-at-tomkaye.com
Scott, Leslie; Two years may be OK if you do not examine the specimen. We have repeatedly seen problems with surface reactions that produces contrast of apparent layers along the surface, and also blobs of dark contrast in the epoxy above the surface when it is over six months old. These epoxies cured fine, and did not delaminate or show other signs of reduced strength, but the specimens had to be prepped over again due to the artifact visible in the microscope.
John Mardinly Intel Corporation
The opinion of this author does not necessarily represent the opinion of Intel corporation.
-----Original Message----- X-from: walck-at-southbaytech.com [mailto:walck-at-southbaytech.com] Sent: Tuesday, August 01, 2006 10:24 PM To: Mardinly, John
Leslie,
I have used batches of Epo-Tek 353ND that I purchased for longer than two years. I started to increase the amount of harder (as measured by drops) when it got older. Actually, what I did was decrease the number of resin drops and kept the hardner drops to 1.
Finally I mixed what was left and cured it on a hot plate. Warning, if you do this, use a big enough container to catch the boil out. I destroyed the surface of a good hot plate.
If you store it at room temp and not in direct sunlight, I think that you can go about 2 years. After that replace it. I did not see a fall off in the adhesion capability of the epoxy as it aged.
In a past life, I've used the G1, with the same results. Since I believe that it is the same stuff as the 353-ND, I will not say any more about it.
SBT does not sell the Epo-Tek 353-ND, we refer our customers to Epoxy Technology, Inc whose website is www.epotek.com.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
-------- Original Message -------- } From: lkrupp-at-us.ibm.com } Sent: Tuesday, August 01, 2006 5:09 PM } To: Walck-at-SouthBayTech.com } Subject: [Microscopy] quality/shelf life of G-1 epoxy, epo-tek, etc. } } ------------------------------------------------------------------------ ---- } 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 listers- } } I have been curious for some time now about how the quality/age of a } certain batch of G-1 (or its equivalents) affects cross-sectioning } properties for TEM samples. The shelf life listed on the small bottles } purchased from Gatan indicate 1 year, as well as the larger bottles from } epo-tek. Has anyone been able to relate poor adhesion, poor ion-milling, } etc. to the age or quality of the epoxy? I know people who have used the } same bottles of epo-tek for years with no problems. } } Thanks, } Leslie } } Leslie Krupp (Thompson) } IBM Almaden Research } 650 Harry Road, K19/D1 } San Jose, CA 95120-6099 } (408) 927-3856 } } ==============================Original Headers============================== } 4, 29 -- From lkrupp-at-us.ibm.com Tue Aug 1 16:05:11 2006 } 4, 29 -- Received: from e4.ny.us.ibm.com (e4.ny.us.ibm.com [32.97.182.144]) } 4, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k71L5Avb024845 } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 16:05:10 -0500 } 4, 29 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) } 4, 29 -- by e4.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k71L56XP011677 } 4, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=FAIL) } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:07 -0400 } 4, 29 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) } 4, 29 -- by d01relay02.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k71L56cA168446 } 4, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 } 4, 29 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) } 4, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k71L56op020767 } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 } 4, 29 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) } 4, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k71L56SH020745 } 4, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 1 Aug 2006 17:05:06 -0400 } 4, 29 -- To: Microscopy-at-microscopy.com } 4, 29 -- Subject: quality/shelf life of G-1 epoxy, epo-tek, etc. } 4, 29 -- MIME-Version: 1.0 } 4, 29 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 } 4, 29 -- Message-ID: {OF9A56A70D.BC20401E-ON852571BD.00702791-882571BD.0073D168-at-us.ibm.com} } 4, 29 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} } 4, 29 -- Date: Tue, 1 Aug 2006 14:05:03 -0700 } 4, 29 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF269 | June 22, 2006) at } 4, 29 -- 08/01/2006 17:05:06, } 4, 29 -- Serialize complete at 08/01/2006 17:05:06 } 4, 29 -- Content-Type: text/plain; charset="US-ASCII" } ==============================End of - Headers==============================
==============================Original Headers============================== 5, 20 -- From walck-at-southbaytech.com Wed Aug 2 00:24:03 2006 5, 20 -- Received: from smtp14.safesecureweb.com (smtp14.safesecureweb.com [66.241.211.108]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k725O26x011759 5, 20 -- for {microscopy-at-microscopy.com} ; Wed, 2 Aug 2006 00:24:02 -0500 5, 20 -- Received: from mail15.safesecureweb.com (ironhide.ad.safesecureweb.com [192.168.2.180]) 5, 20 -- by smtp14.safesecureweb.com (Spam Firewall) with ESMTP 5, 20 -- id B2B6F15BBCF2; Wed, 2 Aug 2006 01:24:01 -0400 (EDT) 5, 20 -- MIME-Version: 1.0 5, 20 -- Date: Wed, 2 Aug 2006 01:23:40 -0400 5, 20 -- Content-Type: text/plain; 5, 20 -- charset=iso-8859-1 5, 20 -- Subject: re: [Microscopy] quality/shelf life of G-1 epoxy, epo-tek, etc. 5, 20 -- From: Scott Walck {walck-at-southbaytech.com} 5, 20 -- Reply-To: Walck-at-southbaytech.com 5, 20 -- To: {lkrupp-at-us.ibm.com} , {microscopy-at-microscopy.com} 5, 20 -- CC: 5, 20 -- Message-ID: {5b68b38b80844900b11aa30fb3f195ea-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 k725O26x011759 ==============================End of - Headers==============================
==============================Original Headers============================== 23, 30 -- From john.mardinly-at-intel.com Sun Aug 6 22:40:08 2006 23, 30 -- Received: from azsmga101-1.ch.intel.com (mga03.intel.com [143.182.124.21]) 23, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k773e8bn005695 23, 30 -- for {Microscopy-at-msa.microscopy.com} ; Sun, 6 Aug 2006 22:40:08 -0500 23, 30 -- Received: from azsmga001.ch.intel.com ([10.2.17.19]) 23, 30 -- by azsmga101-1.ch.intel.com with ESMTP; 06 Aug 2006 20:40:05 -0700 23, 30 -- Received: from scsmsx332.sc.intel.com (HELO scsmsx332.amr.corp.intel.com) ([10.3.90.6]) 23, 30 -- by azsmga001.ch.intel.com with ESMTP; 06 Aug 2006 20:38:47 -0700 23, 30 -- X-IronPort-AV: i="4.07,216,1151910000"; 23, 30 -- d="scan'208"; a="99391439:sNHT143958536988" 23, 30 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.211); 23, 30 -- Sun, 6 Aug 2006 20:38:47 -0700 23, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 23, 30 -- Content-class: urn:content-classes:message 23, 30 -- MIME-Version: 1.0 23, 30 -- Content-Type: text/plain; 23, 30 -- charset="us-ascii" 23, 30 -- Subject: RE: [Microscopy] re: quality/shelf life of G-1 epoxy, epo-tek, etc. 23, 30 -- Date: Sun, 6 Aug 2006 20:38:45 -0700 23, 30 -- Message-ID: {1DF4C4D62339DB4C9C98DF04213995B6B94D79-at-scsmsx413.amr.corp.intel.com} 23, 30 -- X-MS-Has-Attach: 23, 30 -- X-MS-TNEF-Correlator: 23, 30 -- Thread-Topic: [Microscopy] re: quality/shelf life of G-1 epoxy, epo-tek, etc. 23, 30 -- thread-index: Aca18/KSMdelTbaSTcacM7fxAxOYOAD3nDZQ 23, 30 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 23, 30 -- To: {walck-at-southbaytech.com} 23, 30 -- Cc: {Microscopy-at-msa.microscopy.com} 23, 30 -- X-OriginalArrivalTime: 07 Aug 2006 03:38:47.0492 (UTC) FILETIME=[FD8F1040:01C6B9D2] 23, 30 -- Content-Transfer-Encoding: 8bit 23, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k773e8bn005695 ==============================End of - Headers==============================
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Email: polo-at-cameron.net Name: doug
Title-Subject: [Filtered] Acridine Orange and wet mounts
Question: I would be most grateful for any help in formulating an Acridine Orange stain which I can use on blood wet mounts. Any suggestions will be most appreciated. thanks.
I'm a PhD student from the catholic university of Leuven, Belgium and I want to do a TEM on a blend of 2 polymers, filled with particles. The problem is that I have no expercience with this so that's why I send this message. Can somebody help me with my problem? I don't know how I have to prepare my sample. I will give some more information. First I want to put my sample in an oven under N2 for about 7 hours at 220°C. Thereafter, the sample must be cooled down very quickly to freeze the microstructure. After his, I would like to take a TEM picture of the structure. How do I have to prepare my sample to do this? If somebody can explain this in detail, thanks a lot!!
ITEM 1: Tutorial videos from M&M 2006 in Chicago will not be available for about two months. I will post a notice on this list when they are ready for sale.
ITEM 2: At the insistence of MSA leadership, prices for ALL videos will go up on August 15, 2006. Orders recieved prior to that date will be billed at the current price. Orders recieved after that date will be billed at the new prices.
Greg Erdos, MSA Video Wrangler -- 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, 23 -- From gwe-at-ufl.edu Mon Aug 7 10:44:21 2006 3, 23 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k77FiIrK018434 3, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 7 Aug 2006 10:44:20 -0500 3, 23 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 3, 23 -- (authenticated bits=0) 3, 23 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k77FiDI95030110 3, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 7 Aug 2006 11:44:14 -0400 3, 23 -- Message-ID: {44D75FD2.20503-at-ufl.edu} 3, 23 -- Date: Mon, 07 Aug 2006 11:44:18 -0400 3, 23 -- From: Greg Erdos {gwe-at-ufl.edu} 3, 23 -- Reply-To: gwe-at-ufl.edu 3, 23 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 3, 23 -- X-Accept-Language: en-us, en 3, 23 -- MIME-Version: 1.0 3, 23 -- To: "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 3, 23 -- Subject: Tutorial videos 3, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 23 -- Content-Transfer-Encoding: 7bit 3, 23 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 23 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 23 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 3, 23 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
I'm trying to label the ER of my cell line using ER-Tracker Red (a glibenclamide molecule labelled with BODIP-TR from Molecular Probes). Cells are grown on round coverslips and stained with ER-Tracker and fixed as per the instructions. Unfortunately, I have found that although the ER-Tracker labels the ER it also sticks to the entire coverslip creating a bright red field. Reducing the ER-Tracker concentration reduces the background, but also the ER staining. Blocking with a range of reagents (eg. BSA, FCS, human sera, junk proteins, skim milk etc.) does not seem to reduce this non-specific binding to the coverslip at all.
Does anyone have any advice on how to reduce the background? If so, please let me know.
Thanks in advance, Damien Chong
Molecular Life Sciences Building Gate 8, Victoria Drive The University of Adelaide South Australia 5005
==============================Original Headers============================== 5, 29 -- From damien.chong-at-adelaide.edu.au Mon Aug 7 22:52:21 2006 5, 29 -- Received: from tosh.services.adelaide.edu.au (mailguard-send.adelaide.edu.au [192.43.227.21]) 5, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k783qJ4U008836 5, 29 -- for {Microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 22:52:20 -0500 5, 29 -- Received: from stimpy.services.adelaide.edu.au ([10.0.10.10]) 5, 29 -- by tosh.services.adelaide.edu.au with ESMTP; 08 Aug 2006 13:22:18 +0930 5, 29 -- X-IronPort-Anti-Spam-Filtered: true 5, 29 -- X-IronPort-Anti-Spam-Result: AQAAAFyl10QN 5, 29 -- X-IronPort-AV: i="4.07,220,1151850600"; 5, 29 -- d="scan'208"; a="10210367:sNHT15184281" 5, 29 -- Received: from stimpy.services.adelaide.edu.au (localhost.localdomain [127.0.0.1]) 5, 29 -- by stimpy.services.adelaide.edu.au (8.13.1/8.13.1) with ESMTP id k783qI6P010129 5, 29 -- for {Microscopy-at-Microscopy.Com} ; Tue, 8 Aug 2006 13:22:18 +0930 5, 29 -- Received: (from apache-at-localhost) 5, 29 -- by stimpy.services.adelaide.edu.au (8.13.1/8.13.1/Submit) id k783qHZb010119 5, 29 -- for Microscopy-at-Microscopy.Com; Tue, 8 Aug 2006 13:22:17 +0930 5, 29 -- Received: from molb00325.molecularbiosciences.adelaide.edu.au (molb00325.molecularbiosciences.adelaide.edu.au [129.127.106.33]) 5, 29 -- by webmail.adelaide.edu.au (IMP) with HTTP 5, 29 -- for {dchong01-at-localhost} ; Tue, 8 Aug 2006 13:22:17 +0930 5, 29 -- Message-ID: {1155009137.44d80a71af3be-at-webmail.adelaide.edu.au} 5, 29 -- Date: Tue, 8 Aug 2006 13:22:17 +0930 5, 29 -- From: Damien Chong {damien.chong-at-adelaide.edu.au} 5, 29 -- To: Microscopy-at-microscopy.com 5, 29 -- Subject: Fluorescence: ER-Tracker Red background 5, 29 -- MIME-Version: 1.0 5, 29 -- Content-Type: text/plain; charset=ISO-8859-1 5, 29 -- Content-Transfer-Encoding: 8bit 5, 29 -- User-Agent: Internet Messaging Program (IMP) 3.2.6 5, 29 -- X-Originating-IP: 129.127.106.33 ==============================End of - Headers==============================
some more info would be useful, e.g. is it a bulk specimen or a film, what kind of polymers are used and what kind of particles (soft/hard) of which size are incorporated?
Best regards,
Petra
--------------------------------------- Dr. Petra Wahlbring Goodyear S.A. Technical Center Analytical Test Laboratories L-7750 Colmar-Berg Luxembourg Tel +352 8199 3725 Fax +352 8199 3905 e-mail: petra.wahlbring-at-goodyear.com
- May Contain Confidential and/or Proprietary Information. May not be copied or disseminated without the expressed written consent of The Goodyear Tire & Rubber Company. -
Steven.Vandebril-at- cit.kuleuven.be To 08/07/06 04:20 PM petra.wahlbring-at-goodyear.com cc
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Hi all,
I'm a PhD student from the catholic university of Leuven, Belgium and I want to do a TEM on a blend of 2 polymers, filled with particles. The problem is that I have no expercience with this so that's why I send this message. Can somebody help me with my problem? I don't know how I have to prepare my sample. I will give some more information. First I want to put my sample in an oven under N2 for about 7 hours at 220°C. Thereafter, the sample must be cooled down very quickly to freeze the microstructure. After his, I would like to take a TEM picture of the structure. How do I have to prepare my sample to do this? If somebody can explain this in detail, thanks a lot!!
I will give some more information about my sample. Tha particles that I use are carbon black particles with a diameter of 20 nm, so TEM is actually the only option to visualize them. The blend is made of PMMA and PaMSAN, a phase separating blend. Thank you for your reactions!
Citeren petra.wahlbring-at-goodyear.com:
} Dear Steve, } } some more info would be useful, e.g. is it a bulk specimen or a film, } what } kind of polymers are used and what kind of particles (soft/hard) of } which } size are incorporated? } } Best regards, } } Petra } } --------------------------------------- } Dr. Petra Wahlbring } Goodyear S.A. Technical Center } Analytical Test Laboratories } L-7750 Colmar-Berg } Luxembourg } Tel +352 8199 3725 } Fax +352 8199 3905 } e-mail: petra.wahlbring-at-goodyear.com } } - May Contain Confidential and/or Proprietary Information. May not } be } copied or disseminated without the expressed written consent of The } Goodyear Tire & Rubber Company. - } } } } } } Steven.Vandebril-at- } } cit.kuleuven.be } } } To } 08/07/06 04:20 PM petra.wahlbring-at-goodyear.com } } } cc } } } Please respond to } Subject } Steven.Vandebril-at- [Microscopy] TEM: sample } } cit.kuleuven.be preparation polymer blend } } } } } } } } } } } } } } } } } } } } } --------------------------------------------------------------------- ------- } } 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'm a PhD student from the catholic university of Leuven, Belgium } and } I want to do a TEM on a blend of 2 polymers, filled with particles. } The problem is that I have no expercience with this so that's why I } send this message. Can somebody help me with my problem? I don't } know } how I have to prepare my sample. I will give some more information. } First I want to put my sample in an oven under N2 for about 7 hours } at } 220°C. Thereafter, the sample must be cooled down very quickly to } freeze the microstructure. After his, I would like to take a TEM } picture of the structure. How do I have to prepare my sample to do } this? If somebody can explain this in detail, thanks a lot!! } } Kind regards, } } Steven } } } Disclaimer: http://www.kuleuven.be/cwis/email_disclaimer.htm } } } ==============================Original } Headers============================== } 4, 30 -- From Steven.Vandebril-at-cit.kuleuven.be Mon Aug 7 09:16:42 } 2006 } 4, 30 -- Received: from thumbler.kulnet.kuleuven.ac.be } (thumbler.kulnet.kuleuven.ac.be [134.58.240.45]) } 4, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with } ESMTP id k77EGfwc006531 } 4, 30 -- for {microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 } 09:16:41 -0500 } 4, 30 -- Received: from localhost (localhost [127.0.0.1]) } 4, 30 -- by thumbler.kulnet.kuleuven.ac.be (Postfix) with } ESMTP } id 44C45138148 } 4, 30 -- for {microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 } 16:16:40 +0200 (CEST) } 4, 30 -- Received: from smtp02.kuleuven.be } (lepidus.kulnet.kuleuven.ac.be } [134.58.240.72]) } 4, 30 -- by thumbler.kulnet.kuleuven.ac.be (Postfix) with } ESMTP } id 64D61137E8B } 4, 30 -- for {microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 } 16:16:38 +0200 (CEST) } 4, 30 -- Received: from smtp02.kuleuven.be (localhost.localdomain } [127.0.0.1]) } 4, 30 -- by smtp02.kuleuven.be (Postfix) with ESMTP id } 41B822CAA9B; } 4, 30 -- Mon, 7 Aug 2006 16:16:38 +0200 (CEST) } 4, 30 -- Received: from localhost (webmail2.cc.kuleuven.ac.be } [134.58.242.4]) } 4, 30 -- by smtp02.kuleuven.be (Postfix) with ESMTP id } 2E1A52CA9F6 } 4, 30 -- for {microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 } 16:16:38 +0200 (CEST) } 4, 30 -- Received: from dhcp194.cit.kuleuven.be } (dhcp194.cit.kuleuven.be } [10.33.63.194]) } 4, 30 -- by webmail2.kuleuven.be (IMP) with HTTP } 4, 30 -- for {u0050286-at-127.0.0.1} ; Mon, 7 Aug 2006 } 16:16:39 } +0200 } 4, 30 -- Message-ID: {1154960199.44d74b4737263-at-webmail2.kuleuven.be} } 4, 30 -- Date: Mon, 7 Aug 2006 16:16:39 +0200 } 4, 30 -- From: Steven Vandebril {Steven.Vandebril-at-cit.kuleuven.be} } 4, 30 -- To: microscopy-at-microscopy.com } 4, 30 -- Subject: TEM: sample preparation polymer blend } 4, 30 -- MIME-Version: 1.0 } 4, 30 -- Content-Type: text/plain; charset=ISO-8859-1 } 4, 30 -- Content-Transfer-Encoding: 8bit } 4, 30 -- User-Agent: Internet Messaging Program (IMP) 3.2-cvs } 4, 30 -- X-Originating-IP: 10.33.63.194 } 4, 30 -- X-Virus-Scanned: by KULeuven Antivirus Cluster } ==============================End of - } Headers============================== } } }
-- Steven Vandebril Lab of applied rheology and polymer processing Chemical Engineering Department - K.U.Leuven W. de Croylaan 46 3001 Heverlee Tel.:0476 231925 Email:steven.vandebril-at-cit.kuleuven.be
with a soft filler like CB, I think that ultra-microtome cutting of thin sections (at room temperature since Tg's of both polymers are } 100°C) is the method of choice, followed by a Ruthenium Tetroxide staining in order to achieve a contrast between the PaMSAN (stained by RuO4 due to the aromatic ring in the a-methylstyrene) and the PMMA (less stained by RuO4). If you need more detailed information on RuO4 staining, you can contact me again.
With best regards,
Petra
P.S. This report might be of interest for you: http://sunsite.online.globule.org/iupac/publications/pac/1998/pdf/7008x1547.pdf
--------------------------------------- Dr. Petra Wahlbring Goodyear S.A. Technical Center Analytical Test Laboratories L-7750 Colmar-Berg Luxembourg Tel +352 8199 3725 Fax +352 8199 3905 e-mail: petra.wahlbring-at-goodyear.com
- May Contain Confidential and/or Proprietary Information. May not be copied or disseminated without the expressed written consent of The Goodyear Tire & Rubber Company. -
Steven Vandebril {Steven.Vandebril -at-cit.kuleuven.be} To petra.wahlbring-at-goodyear.com 08/08/06 09:08 AM cc Microscopy-at-Microscopy.Com Subject Re: [Microscopy] TEM: sample preparation polymer blend
Hello,
I will give some more information about my sample. Tha particles that I use are carbon black particles with a diameter of 20 nm, so TEM is actually the only option to visualize them. The blend is made of PMMA and PaMSAN, a phase separating blend. Thank you for your reactions!
Citeren petra.wahlbring-at-goodyear.com:
} Dear Steve, } } some more info would be useful, e.g. is it a bulk specimen or a film, } what } kind of polymers are used and what kind of particles (soft/hard) of } which } size are incorporated? } } Best regards, } } Petra } } --------------------------------------- } Dr. Petra Wahlbring } Goodyear S.A. Technical Center } Analytical Test Laboratories } L-7750 Colmar-Berg } Luxembourg } Tel +352 8199 3725 } Fax +352 8199 3905 } e-mail: petra.wahlbring-at-goodyear.com } } - May Contain Confidential and/or Proprietary Information. May not } be } copied or disseminated without the expressed written consent of The } Goodyear Tire & Rubber Company. - } } } } } } Steven.Vandebril-at- } } cit.kuleuven.be } } } To } 08/07/06 04:20 PM petra.wahlbring-at-goodyear.com } } } cc } } } Please respond to } Subject } Steven.Vandebril-at- [Microscopy] TEM: sample } } cit.kuleuven.be preparation polymer blend } } } } } } } } } } } } } } } } } } } } } --------------------------------------------------------------------- ------- } } 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'm a PhD student from the catholic university of Leuven, Belgium } and } I want to do a TEM on a blend of 2 polymers, filled with particles. } The problem is that I have no expercience with this so that's why I } send this message. Can somebody help me with my problem? I don't } know } how I have to prepare my sample. I will give some more information. } First I want to put my sample in an oven under N2 for about 7 hours } at } 220°C. Thereafter, the sample must be cooled down very quickly to } freeze the microstructure. After his, I would like to take a TEM } picture of the structure. How do I have to prepare my sample to do } this? If somebody can explain this in detail, thanks a lot!! } } Kind regards, } } Steven } } } Disclaimer: http://www.kuleuven.be/cwis/email_disclaimer.htm } } } ==============================Original } Headers============================== } 4, 30 -- From Steven.Vandebril-at-cit.kuleuven.be Mon Aug 7 09:16:42 } 2006 } 4, 30 -- Received: from thumbler.kulnet.kuleuven.ac.be } (thumbler.kulnet.kuleuven.ac.be [134.58.240.45]) } 4, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with } ESMTP id k77EGfwc006531 } 4, 30 -- for {microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 } 09:16:41 -0500 } 4, 30 -- Received: from localhost (localhost [127.0.0.1]) } 4, 30 -- by thumbler.kulnet.kuleuven.ac.be (Postfix) with } ESMTP } id 44C45138148 } 4, 30 -- for {microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 } 16:16:40 +0200 (CEST) } 4, 30 -- Received: from smtp02.kuleuven.be } (lepidus.kulnet.kuleuven.ac.be } [134.58.240.72]) } 4, 30 -- by thumbler.kulnet.kuleuven.ac.be (Postfix) with } ESMTP } id 64D61137E8B } 4, 30 -- for {microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 } 16:16:38 +0200 (CEST) } 4, 30 -- Received: from smtp02.kuleuven.be (localhost.localdomain } [127.0.0.1]) } 4, 30 -- by smtp02.kuleuven.be (Postfix) with ESMTP id } 41B822CAA9B; } 4, 30 -- Mon, 7 Aug 2006 16:16:38 +0200 (CEST) } 4, 30 -- Received: from localhost (webmail2.cc.kuleuven.ac.be } [134.58.242.4]) } 4, 30 -- by smtp02.kuleuven.be (Postfix) with ESMTP id } 2E1A52CA9F6 } 4, 30 -- for {microscopy-at-microscopy.com} ; Mon, 7 Aug 2006 } 16:16:38 +0200 (CEST) } 4, 30 -- Received: from dhcp194.cit.kuleuven.be } (dhcp194.cit.kuleuven.be } [10.33.63.194]) } 4, 30 -- by webmail2.kuleuven.be (IMP) with HTTP } 4, 30 -- for {u0050286-at-127.0.0.1} ; Mon, 7 Aug 2006 } 16:16:39 } +0200 } 4, 30 -- Message-ID: {1154960199.44d74b4737263-at-webmail2.kuleuven.be} } 4, 30 -- Date: Mon, 7 Aug 2006 16:16:39 +0200 } 4, 30 -- From: Steven Vandebril {Steven.Vandebril-at-cit.kuleuven.be} } 4, 30 -- To: microscopy-at-microscopy.com } 4, 30 -- Subject: TEM: sample preparation polymer blend } 4, 30 -- MIME-Version: 1.0 } 4, 30 -- Content-Type: text/plain; charset=ISO-8859-1 } 4, 30 -- Content-Transfer-Encoding: 8bit } 4, 30 -- User-Agent: Internet Messaging Program (IMP) 3.2-cvs } 4, 30 -- X-Originating-IP: 10.33.63.194 } 4, 30 -- X-Virus-Scanned: by KULeuven Antivirus Cluster } ==============================End of - } Headers============================== } } }
-- Steven Vandebril Lab of applied rheology and polymer processing Chemical Engineering Department - K.U.Leuven W. de Croylaan 46 3001 Heverlee Tel.:0476 231925 Email:steven.vandebril-at-cit.kuleuven.be
MANAGER IN CRYOELECTRON MICROSCOPY NEW YORK STRUCTURAL BIOLOGY CENTER
The New York Structural Biology Center seeks a technical manager for a high-end, state or the art facility in Cryoelectron Microscopy. The facility includes four cryoelectron microscopes at 120, 200(2), and 300 kV, the last with a liquid-helium stage and an energy filter, as well as all necessary ancillary equipment. The facility is used by investigators from ten New York academic research institutions on a broad range of biological targets employing all of three CryoEM methodologies: tomography, single-particle analysis, and crystallography. The appointee will work with manufacturers in maintaining optimal performance of the instruments and will work with scientists in pursuing a broad range of biomedical applications, some of which are likely to require new developments. The appointee will supervise an existing team of technical staff with expertise in sample preparation and computational analysis. A strong technical background in electron microscopy is essential including familiarity with biological applications. Compensation will be appropriate and competitive for the candidate's level of experience and training. Send a biographical sketch, a brief statement of previous research experience, together with names and addresses of three individuals who can provide letters of recommendation. Applications should be sent as soon as possible to: David Stokes, at stokes-at-nysbc.org. For more information, see http://www.nysbc.org/facilities/CEM
-- David L. Stokes New York Structural Biology Center tel: 212-939-0660 x116 http://nysbc.org/ fax: 646-219-0300
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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 wim.vandenbroeck-at-UGent.be as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: wim.vandenbroeck-at-UGent.be Name: Wim Van den Broeck
Organization: Department of Morphology, Ghent University
Title-Subject: [Filtered] TEM lab: organization
Question: Dear colleagues,
In the next months, I will have the budget to set up a TEM lab with the following instruments: EM tissue processor, knife maker, ultramicrotome and table, EM trimming instrument, EM staining instrument. In Belgium, as far as I know, Leica is the only company that distributes these instruments. Please, can you tell me if there are other companies, and what the the proís and conís are of the different instruments / different companies. A second question deals with the organization of the lab. I have a room of about 30 m¾ at my disposal with a solid lab table in the centre and two units with fumehoods at one side. Can you give my some tips how to organize the lab as efficiently as possible ?
Thanks in advance for your kind help. Wim Van den Broeck.
I am unable to maintain a high vacuum and the failure message on my Quanta 200 SEM reads that there is a gauge failure and the source is the vacuum. I suspect that it is the Penning gauge and would like to clean it. It is a Pfeiffer model that operates at approximately 2kV. The FEI service rep. refers to this particular gauge as being 'active'. Has anyone had success at dismantling and cleaning this type of gauge. The cost of having it serviced is considerable. Any advice or suggestions on how to proceed with cleaning, etc. can be made to me directly and 'off-line'.
Regards Deb
Debra L. Moreau, Ph.D. Electron Microscopy and Imaging Agriculture and Agri-Food Canada/ Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902 679-6513 Facsimile/Télécopieur: 902 679-2311 32 Main Street Kentville, Nova Scotia B4N 1J5 moreaud-at-agr.gc.ca
==============================Original Headers============================== 6, 26 -- From MoreauD-at-AGR.GC.CA Wed Aug 9 08:33:53 2006 6, 26 -- Received: from agrpazsmtp2.agr.gc.ca (agrpazsmtp2.agr.gc.ca [192.197.71.137]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k79DXrfc005335 6, 26 -- for {Microscopy-at-Microscopy.Com} ; Wed, 9 Aug 2006 08:33:53 -0500 6, 26 -- Received: from onncrxcn4.AGR.GC.CA ([192.168.122.1]) 6, 26 -- by agrpazsmtp2.agr.gc.ca (8.13.6/8.13.3) with ESMTP id k79DUDLs003540 6, 26 -- for {Microscopy-at-Microscopy.Com} ; Wed, 9 Aug 2006 09:30:13 -0400 6, 26 -- Received: from onncrxms3.AGR.GC.CA ([10.117.15.30]) by onncrxcn4.AGR.GC.CA with Microsoft SMTPSVC(5.0.2195.6713); 6, 26 -- Wed, 9 Aug 2006 09:33:34 -0400 6, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 6, 26 -- content-class: urn:content-classes:message 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; 6, 26 -- charset="iso-8859-1" 6, 26 -- Subject: Re: cleaning a Penning gauge 6, 26 -- Date: Wed, 9 Aug 2006 09:32:56 -0400 6, 26 -- Message-ID: {E91ED609E45D144C99D56492ABDF74C30352215E-at-onncrxms3.agr.gc.ca} 6, 26 -- X-MS-Has-Attach: 6, 26 -- X-MS-TNEF-Correlator: 6, 26 -- Thread-Topic: Re: cleaning a Penning gauge 6, 26 -- Thread-Index: Aca7uFLDcRLeCq7RT7WggBMv1EaM7A== 6, 26 -- From: "Moreau, Debbie" {MoreauD-at-AGR.GC.CA} 6, 26 -- To: {Microscopy-at-Microscopy.Com} 6, 26 -- X-OriginalArrivalTime: 09 Aug 2006 13:33:34.0340 (UTC) FILETIME=[69676C40:01C6BBB8] 6, 26 -- Content-Transfer-Encoding: 8bit 6, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k79DXrfc005335 ==============================End of - Headers==============================
Dear Wim First, let me congratulate you. In these times of slashed budgets its very good to hear of someone being able to set up a lab, rather than dismantle one! Try to get your hands on a copy of volume 4 of the "Practical Methods in Electron Microscopy" series edited by Audrey M. Glauert: Design of the Electron Microscope Laboratory" by Ronald H. Alderson. I think that its out of print (original copyright was 1975) but it is still an excellent guide and addresses the set up of labs of varying sizes.
Lee Cohen-Gould -- 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 Wed Aug 9 08:35:36 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 k79DZYMU008686 2, 24 -- for {microscopy-at-microscopy.com} ; Wed, 9 Aug 2006 08:35:35 -0500 2, 24 -- Received: from mpx2.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 k79DZVHA024590 2, 24 -- for {microscopy-at-microscopy.com} ; Wed, 9 Aug 2006 09:35:31 -0400 (EDT) 2, 24 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 2, 24 -- by mpx2.med.cornell.edu 2, 24 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 2, 24 -- with ESMTPA id {0J3Q00GRXGF12240-at-mpx2.med.cornell.edu} for 2, 24 -- microscopy-at-microscopy.com; Wed, 09 Aug 2006 09:35:25 -0400 (EDT) 2, 24 -- Date: Wed, 09 Aug 2006 09:26:41 -0400 2, 24 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 2, 24 -- Subject: Re: [Microscopy] viaWWW: TEM lab: organization 2, 24 -- In-reply-to: {200608091315.k79DF5fv028307-at-ns.microscopy.com} 2, 24 -- Sender: lcgould-at-med.cornell.edu 2, 24 -- To: wim.vandenbroeck-at-UGent.be, 2, 24 -- Microscopy Listserver {microscopy-at-microscopy.com} 2, 24 -- Message-id: {p06230902c0ff91f46e6a-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: {200608091315.k79DF5fv028307-at-ns.microscopy.com} 2, 24 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.8.9.62433 ==============================End of - Headers==============================
Dear All, is there anybody out there who has some layouts for the EDAX 184 pre-amp? I got an EDAX ECON detector which will be attached to an EDAX 9100. I need to know for which pots I need for trimming the pre-amp. There are two pots, one is black, the other gray... I also need a hint where to find the Gain and the Zero pots on the ADC board.
Listers: We would like to upgrade our color imaging system for histological slides - does anybody have any camera systems that they hate/love and could comment on? The Nikon CoolPix seems to be quite popular; how is the software? Can it be convinced to talk to MetaMorph? Has anybody used the MotiCam 2000? It looks spiffy, but I didn't see any references to it in the archives. What about the Orion astronomy camera? We have an old Kodak now that is no longer supported and the software is kind of hopeless, so the users are agitating for an upgrade. We don't want to spend a ton of money, of course...this is just for plain old histo.
Dear Deb, If your Penning gauge is the usual type: cold, with a strong magnet wrapped around it, then you need to take it off the SEM, disassemble it and clean all the bits inside. Whiskers form on the inside over time and these interfere with the ion flow that measures the vacuum. I cannot be more specific because I don't have one of that particular brand. The Edwards ones I have consist of several plates and a needle up the middle. The ion flow is between the needle and plates. The magnet extends the range of the vacuum reading. I just clean the plates and needle and inside of the cylindrical body with fine emery paper and metal polish and wash everything clean with acetone or lab alcohol, dry and re-assemble. Usually works. If you have the instruction manual that comes with the gauge, it may tell you how to clean it, or the SEM maintenance instructions. I hope this helps. Please feel free to contact me if you have any other questions. 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: {MoreauD-at-AGR.GC.CA} To: {mager-at-interchange.ubc.ca} Sent: Wednesday, August 09, 2006 6:39 AM
Attendance is limited! Additional morning presentations are welcome- please contact John Donovan directly.
EDS Spectrum Imaging and Low Voltage (High Resolution) Analysis, University of Oregon, Eugene, OR
3 day workshop by Dale Newbury (National Institute of Standards & Technology), September, 12-14, 2006
Morning 1 talk: "Silicon Drift Detectors: Energy Dispersive X-ray Spectrometry and X-ray Spectrum Imaging at Output Count Rates Above 100 kHz, and What to Do with All This Data"
Afternoon Demos: Spectral Imaging and Mapping with a conventional EDS (Oxford and Thermo) Bruker Quad SDD and Thermo SDD doing high speed x-ray spectrum image mapping on an FEI Quanta SEM LISPIX hands-on software lab, with pre-recorded x-ray spectrum image data sets.
Morning 2 talk: "Challenges to Successful EDS X-ray Microanalysis in the Low Beam Energy (E0 { 5 keV) Regime"
Afternoon demos: Low Beam Energy SEM/EDS on selected specimens, e.g., SiC, BaTiO3, etc. Demonstration of conventional WDS in the low beam energy regime with BaTiO3 scanning the TiL-O K and BaM spectral regions.
Morning 3 talk: "The Perils of Automatic EDS Analysis: Blunders in Automated Peak Identification of Major, Minor, and Trace Constituents and the Reality of Standardless Analysis"
Afternoon demo: Testing Automatic Peak ID and Standardless Analysis with commercial software in the Laboratory User suggestions for testing (bring your own samples)
Check this page for registration and further details: http://epmalab.uoregon.edu/eds_workshop.htm
Respond to John Donovan (donovan-at-uoregon.edu) to confirm your participation. Registration is required by September 1, 2006. More to come...
John J. Donovan donovan-at-uoregon.edu University of Oregon (541) 346-4632 (office) 1260 Franklin Blvd (541) 346-4655 (probe) Eugene, OR (541) 346-4778 (SEM) 97403-1272 (541) 346-4692 (FAX) Lab Web: http://epmalab.uoregon.edu/
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dirk.durinck-at-mtm.kuleuven.be) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, August 10, 2006 at 04:46:59 ---------------------------------------------------------------------------
I am a PhD student at the Catholic University of Leuven (Belgium) in material sciences with a question about microscopy.
One of my collegues is exploring the capabilities of a CSLM to observe in-situ the high temperature behaviour of refractory materials. The material is not transparent to the laser light, so he can only see the surface of the sample. He is using the cslm because our institute does not have a optical microscope with a heating stage.
A problem he faces is that the image quality deteriorates quite rapidly when the sample reaches high temperatures (1600ƒC). The explanation we are thinking of, is that the sample roughens during the experiment. Measurement has shown that before the tests the roughness was about 0.2µm; after the tests is was 5µm. The small focus plane of the CSLM could thus cause blurring of the image.
In order to elaborate this explanation, we would like to have an idea about the axial (depth, vertical) resolution of the CSLM equipment and other imaging techniques (LOM, SEM). Can someone please suggest a good reference where we can find such data?
Usual failure mode of CCIG gages in SEM and FIB tools is dielectric deposition from hydrocarbon oils and precursor gases on electrodes of the sensor tube, and removal of these deposits fully restores work resource of gage. So far I cleaned hundreds (literally) of Edwards and Varian CCIG sensors by following procedure:
1. Dismount CCIG from the tool, remove magnet, and carefully take sensor tube apart.
2. Soak all the metal parts of the sensor tube overnight in 50/50 solution of Micro 90 and DI or distilled water. Micro 90 is available from Cole Parmer http://www.coleparmer.ca/catalog/product_view.asp?sku=1810001&pfx=EW and if you do not have DI then distilled water from supermarket works. Avoid tap water though.
3. In the morning sonic metal parts for 10 min. in the same solution in which they were soaking. At this point all deposits would be removed from the electrodes and you should see nice shiny metal surfaces.
4. Thoroughly rinse the parts, dry them (hot plate or oven to speed up), and put the sensor tube together - done.
Micro 90 cleaning is very gentle, does not scratch electrodes, and takes minimum of your own time - you can do other things while gage is soaking or drying.
If you do not mind time investment, cleaning by hand-polishing of the electrodes, which was already suggested, will also work.
The only thing I can not help you with is how to take apart Pfeiffer gage; I did cleaning of only Edwards and Varian gages so far.
The best for you would probably be to pre-clean one or two old gages, which you most likely have laying around the lab, and replace gage on the tool as needed.
Cheers, Valery
-----Original Message----- X-from: MoreauD-at-AGR.GC.CA [mailto:MoreauD-at-AGR.GC.CA] Sent: Wednesday, August 09, 2006 9:35 AM To: vray-at-partbeamsystech.com
Good morning.
I am unable to maintain a high vacuum and the failure message on my Quanta 200 SEM reads that there is a gauge failure and the source is the vacuum. I suspect that it is the Penning gauge and would like to clean it. It is a Pfeiffer model that operates at approximately 2kV. The FEI service rep. refers to this particular gauge as being 'active'. Has anyone had success at dismantling and cleaning this type of gauge. The cost of having it serviced is considerable. Any advice or suggestions on how to proceed with cleaning, etc. can be made to me directly and 'off-line'.
Regards Deb
Debra L. Moreau, Ph.D. Electron Microscopy and Imaging Agriculture and Agri-Food Canada/ Agriculture et Agroalimentaire Canada Telephone/Téléphone: 902 679-6513 Facsimile/Télécopieur: 902 679-2311 32 Main Street Kentville, Nova Scotia B4N 1J5 moreaud-at-agr.gc.ca
==============================Original Headers============================== 6, 26 -- From MoreauD-at-AGR.GC.CA Wed Aug 9 08:33:53 2006 6, 26 -- Received: from agrpazsmtp2.agr.gc.ca (agrpazsmtp2.agr.gc.ca [192.197.71.137]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k79DXrfc005335 6, 26 -- for {Microscopy-at-Microscopy.Com} ; Wed, 9 Aug 2006 08:33:53 -0500 6, 26 -- Received: from onncrxcn4.AGR.GC.CA ([192.168.122.1]) 6, 26 -- by agrpazsmtp2.agr.gc.ca (8.13.6/8.13.3) with ESMTP id k79DUDLs003540 6, 26 -- for {Microscopy-at-Microscopy.Com} ; Wed, 9 Aug 2006 09:30:13 -0400 6, 26 -- Received: from onncrxms3.AGR.GC.CA ([10.117.15.30]) by onncrxcn4.AGR.GC.CA with Microsoft SMTPSVC(5.0.2195.6713); 6, 26 -- Wed, 9 Aug 2006 09:33:34 -0400 6, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 6, 26 -- content-class: urn:content-classes:message 6, 26 -- MIME-Version: 1.0 6, 26 -- Content-Type: text/plain; 6, 26 -- charset="iso-8859-1" 6, 26 -- Subject: Re: cleaning a Penning gauge 6, 26 -- Date: Wed, 9 Aug 2006 09:32:56 -0400 6, 26 -- Message-ID: {E91ED609E45D144C99D56492ABDF74C30352215E-at-onncrxms3.agr.gc.ca} 6, 26 -- X-MS-Has-Attach: 6, 26 -- X-MS-TNEF-Correlator: 6, 26 -- Thread-Topic: Re: cleaning a Penning gauge 6, 26 -- Thread-Index: Aca7uFLDcRLeCq7RT7WggBMv1EaM7A== 6, 26 -- From: "Moreau, Debbie" {MoreauD-at-AGR.GC.CA} 6, 26 -- To: {Microscopy-at-Microscopy.Com} 6, 26 -- X-OriginalArrivalTime: 09 Aug 2006 13:33:34.0340 (UTC) FILETIME=[69676C40:01C6BBB8] 6, 26 -- Content-Transfer-Encoding: 8bit 6, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k79DXrfc005335 ==============================End of - Headers==============================
==============================Original Headers============================== 26, 24 -- From vray-at-partbeamsystech.com Thu Aug 10 11:04:20 2006 26, 24 -- Received: from smtp106.plus.mail.re2.yahoo.com (smtp106.plus.mail.re2.yahoo.com [206.190.53.31]) 26, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7AG4K6i014613 26, 24 -- for {microscopy-at-microscopy.com} ; Thu, 10 Aug 2006 11:04:20 -0500 26, 24 -- Message-Id: {200608101604.k7AG4K6i014613-at-ns.microscopy.com} 26, 24 -- Received: (qmail 64034 invoked from network); 10 Aug 2006 16:04:19 -0000 26, 24 -- Received: from unknown (HELO MEOMobile1) (partbeamsystech-at-200.88.244.199 with login) 26, 24 -- by smtp106.plus.mail.re2.yahoo.com with SMTP; 10 Aug 2006 16:04:19 -0000 26, 24 -- Reply-To: {vray-at-partbeamsystech.com} 26, 24 -- From: "Valery Ray" {vray-at-partbeamsystech.com} 26, 24 -- To: {MoreauD-at-AGR.GC.CA} 26, 24 -- Cc: {microscopy-at-microscopy.com} 26, 24 -- Subject: RE: [Microscopy] Re: cleaning a Penning gauge 26, 24 -- Date: Thu, 10 Aug 2006 12:04:31 -0400 26, 24 -- Organization: PBST / MEO Engineering 26, 24 -- MIME-Version: 1.0 26, 24 -- Content-Type: text/plain; 26, 24 -- charset="iso-8859-1" 26, 24 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 26, 24 -- Thread-Index: Aca7uJOFw7wknLrzSqW8BR6F3OFbbAA2NhNw 26, 24 -- In-Reply-To: {200608091334.k79DYi8Y007350-at-ns.microscopy.com} 26, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 26, 24 -- Content-Transfer-Encoding: 8bit 26, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7AG4K6i014613 ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (cbennett-at-tulane.edu) from on Thursday, August 10, 2006 at 14:45:53 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both cbennett-at-tulane.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Title: cryoSEM of leukemia cell surface (membrane)
Question: I am attempting to visualize the cellular surface of leukemia cells (~10um) after treatment with a peptide using cryoSEM. I am interested in noting the changes in the membrane such as membrane blebbing or the formation of pores. Currently, I am incubating the cells with peptide in RPMI 1640 in the presence of polyK coated slides. After the assay is completed I transfer the slides to fresh RPMI. (I have tried tranferring into water, but the images are of poorer quality - more ice crystals) I perform a 10min sublimation at -95C for the samples. The images that I get vary from day to day and I was wondering if there was a better way to prepare my samples just prior to loading. Thank you.
The University of Missouri is looking for an ASSOCIATE DIRECTOR for a campus-wide light microscopy imaging facility.
Applicants should have experience in some or all of the following areas: * confocal scanning laser microscopy * bright field microscopy * wide field fluorescence microscopy * deconvolution * FRET/FLIM * image processing/analysis * all types of microtomy * immunocytochemistry * in situ hybridization
The Associate Director will be responsible for training users, maintaining instruments and developing protocols for a campus-wide multi-user facility. PhD desirable but not required for individuals with extensive experience. Although an ideal candidate would have experience in all of the areas listed above, candidates with extensive experience in selected areas and who have the desire and capacity to learn the additional areas will be considered. Excellent oral and written communication skills are essential. Experience in a multi-user core facility would be viewed positively. Electron microscopy is not a component of this core facility. Women and minority candidates are especially encouraged to apply. Review of applications will begin immediately and continue until an appropriate candidate is hired. This is a full-time, benefit eligible position. Opportunities for professional development will be encouraged.
The Core's web site can be viewed at www.biotech.missouri.edu/mcc/index.html--|http://www.biotech.missouri.edu/mcc/index.html
Available instrumentation includes: * Zeiss Meta NLO 2 Photon confocal system * BioRad Radiance 2000 confocal system * Leica DMI4000 B and Eppendorf microjection system * Olympus IX70 and Nikon Optiphot 2 widefield systems * Leica stereoscope equipped with epi-fluorescence excitation * Intavis InsituPro VS in situ hybridization robotic system * Autodeblur deconvolution software (Autoquant Inc.). * Metamorph, Imaris Suite, ImagePro image analysis and processing software. * Leica ultramicrotome, cryostat, paraffin microtomes Address applications (CV and 3 letters of reference) or inquires to:
Thomas E. Phillips, Ph.D. Division of Biological Sciences 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 Fri Aug 11 08:59:53 2006 7, 18 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7BDxqY1024585 7, 18 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 11 Aug 2006 08:59:52 -0500 7, 18 -- Received: from um-nsmtpout1.um.umsystem.edu ([209.106.228.53]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 7, 18 -- Fri, 11 Aug 2006 08:59:52 -0500 7, 18 -- Received: from phillips-dell.missouri.edu ([128.206.81.132]) by um-nsmtpout1.um.umsystem.edu over TLS secured channel with Microsoft SMTPSVC(6.0.3790.1830); 7, 18 -- Fri, 11 Aug 2006 08:59:51 -0500 7, 18 -- Message-Id: {6.0.0.22.2.20060811085848.043acb80-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: Fri, 11 Aug 2006 08:59:47 -0500 7, 18 -- To: Microscopy-at-msa.microscopy.com 7, 18 -- From: Tom Phillips {phillipst-at-missouri.edu} 7, 18 -- Subject: Associate Director position LM facility opening 7, 18 -- Mime-Version: 1.0 7, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 7, 18 -- X-OriginalArrivalTime: 11 Aug 2006 13:59:51.0938 (UTC) FILETIME=[6A8D4620:01C6BD4E] ==============================End of - Headers==============================
I have a guy who wants to do diffraction on a protein fibril. We usually look at these things negatively stained with UA and just check for fibrils or no fibrils in the sample. The samples can be kind of variable and junky.
The fibrils are about 12 nm (can this be right?) wide by much longer. I'm not much good with diffraction so I told him I would scout around for someone who could help him.
We are located near the SF bay area. If you could help us with this project, let me know and I will pass along the info.
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============================== 8, 21 -- From jmkrupp-at-ucsc.edu Fri Aug 11 15:47:42 2006 8, 21 -- Received: from smtp-prod-mx1.ucsc.edu (smtp-prod-mx1.ucsc.edu [128.114.125.43]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7BKlg12009685 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 11 Aug 2006 15:47:42 -0500 8, 21 -- Received: from ucsc.edu (cruzmail-fe1.ucsc.edu [128.114.125.5]) 8, 21 -- by smtp-prod-mx1.ucsc.edu (8.13.7/8.13.7) with ESMTP id k7BKlOJv001025 8, 21 -- for {microscopy-at-microscopy.com} ; Fri, 11 Aug 2006 13:47:36 -0700 (PDT) 8, 21 -- Received: from [128.114.25.186] (account jmkrupp-at-ucsc.edu [128.114.25.186] verified) 8, 21 -- by copper.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 8, 21 -- with ESMTPA id 65895025 for microscopy-at-microscopy.com; Fri, 11 Aug 2006 13:47:29 -0700 8, 21 -- Mime-Version: 1.0 8, 21 -- Message-Id: {p06230903c1029bf13108-at-[128.114.25.186]} 8, 21 -- Date: Fri, 11 Aug 2006 13:47:28 -0700 8, 21 -- To: microscopy-at-microscopy.com 8, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 8, 21 -- Subject: Diffraction on protein fibril? 8, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 8, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 8, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 8, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 8, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (sawyert-at-science.oregonstate.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, August 11, 2006 at 10:38:32 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both sawyert-at-science.oregonstate.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Question: I have been working with gfp inserted into plant pathogenic bacterial. My current problem is photo bleaching. The cells have been fixed in para formaldehyde and washed but while trying to capture the image electronically the gfp rapidly bleaches. To quantify data I need an image so any suggestion will be helpful.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gmerkiso-at-iupui.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, August 12, 2006 at 12:09:54 ---------------------------------------------------------------------------
Email: gmerkiso-at-iupui.edu Name: George Merkison
Organization: Indiana Blood Centter
Education: Undergraduate College
Location: Indianapolis,IN
Question: When using a light microscope, I was taught to keep the condenser down for fluids, including urine sediment and cell counts on hemacytometer, and Nagoette chambers. Is this true and is there a reference to show other individuals? I can't find a reference. Thanks for your reply.
Although I understand the principle behind electron diffraction in TEM I have no idea about the kind of informations this technique gives and how to interpret the diffraction pattern. If you could give me a www address which explains that I would be much grateful.
Now a very practical problem could be perhaps used as an example: we have a mixture of aluminosilicate mineral (I have cut 70 nm sections) with approx. 90% of mordenite and 10% quarz. 1) Can I use e diffraction to distinguish both types of particles? (to verify the purity of the powder) 2) What kind of information about the cristal structure can e diffraction give me in this case? 3) Can I detect a change -and which change- to the cristal structure using this technique if the mineral is heat and treated with strong acids? (which actually modifies the structure)
We have also a tilt stage for tomography. Can this bring further informations?
Regards,
Stephane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 8, 18 -- From nizets2-at-yahoo.com Mon Aug 14 06:30:17 2006 8, 18 -- Received: from web37406.mail.mud.yahoo.com (web37406.mail.mud.yahoo.com [209.191.91.138]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7EBUGao010591 8, 18 -- for {microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 06:30:17 -0500 8, 18 -- Received: (qmail 81606 invoked by uid 60001); 14 Aug 2006 11:30:15 -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=rlmt7902ZNRzFTTFfQF7ruMzaWPunDe6Tu9YxDJ2pD9W3I+BsUwz4JFylLSb1NbV5Y1qFqa1jK6YE5BUUAa9EVhASSCrla6W/fKuGhTPt6o7J7paQwTF6sqlqLCbo9YrFe2LwGUS/eGKK8kO3I8ZVczxOMyA1VvnaLJ+c9MHHSw= ; 8, 18 -- Message-ID: {20060814113015.81604.qmail-at-web37406.mail.mud.yahoo.com} 8, 18 -- Received: from [80.122.101.102] by web37406.mail.mud.yahoo.com via HTTP; Mon, 14 Aug 2006 04:30:15 PDT 8, 18 -- Date: Mon, 14 Aug 2006 04:30:15 -0700 (PDT) 8, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 18 -- Subject: need infos about e diffraction in TEM 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==============================
The example you give, distinguishing between two phases of different structure in a single sample, is the most common use of electron diffraction. This is essentially a fingerprinting of a structure using a measure of angles and spacings provided by the pattern and fitting these to the geometry of a known unit cell. Strictly speaking, this can only reject a candidate phase by failure to fit, because you aren't really proving the presence of a particular structure, just showing that it's a plausible match to the data. This is nicely suited to your case of only having two structures to choose from.
If you've never done this before, you should try it first on a single-phase material (for example a finely ground silicon). This will give you some practice at orienting a zone axis (you will need to use a double-tilt holder, so this will eliminate the tomography holder unless yours can tilt on two axes). Once you have done this you will also know the camera constant of the microscope for a given camera length, which will provide essential information for distinguishing phases (relates the spacing in A to the distance of a reflection from the pattern origin).
One thing working to your advantage will be the large spacings present in mordenite and the absence of any very large spacings in (alpha) quartz. Because of this, any time you see a reflection indicating a spacing greater than about 4.5 A it must be the mordenite.
Structural changes may be difficult to detect depending on what they are. They would have to involve a fairly large change in the size and shape of the unit cell in order to detect with 'spot pattern' diffraction. If you have a material amenable to convergent beam diffraction you can have a lot more sensitivity to structural changes but your sample must be quite beam stable and have relatively small unit cell (neither is likely to be the case for your mordenite).
As far as references go, the old standard Hirsch et al (Electron Microscopy of Thin Crystals) is pretty good on basic spot pattern indexing - see chapter 5 and appendices 5 and 6 which show some worked examples. In fact, any TEM textbook should have at least some discussion of how to acquire and index spot patterns.
Good Luck, Wharton
************************************************************* Wharton Sinkler, PhD. UOP LLC 50 E. Algonquin Rd. Des Plaines IL 60017-5017 mailto: wharton.sinkler-at-uop.com tel 847-391-3878 fax 847-391-3719
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Monday, August 14, 2006 6:36 AM To: Sinkler, Wharton
Dear listers,
Although I understand the principle behind electron diffraction in TEM I have no idea about the kind of informations this technique gives and how to interpret the diffraction pattern. If you could give me a www address which explains that I would be much grateful.
Now a very practical problem could be perhaps used as an example: we have a mixture of aluminosilicate mineral (I have cut 70 nm sections) with approx. 90% of mordenite and 10% quarz. 1) Can I use e diffraction to distinguish both types of particles? (to verify the purity of the powder) 2) What kind of information about the cristal structure can e diffraction give me in this case? 3) Can I detect a change -and which change- to the cristal structure using this technique if the mineral is heat and treated with strong acids? (which actually modifies the structure)
We have also a tilt stage for tomography. Can this bring further informations?
Regards,
Stephane
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==============================Original Headers============================== 8, 18 -- From nizets2-at-yahoo.com Mon Aug 14 06:30:17 2006 8, 18 -- Received: from web37406.mail.mud.yahoo.com (web37406.mail.mud.yahoo.com [209.191.91.138]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7EBUGao010591 8, 18 -- for {microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 06:30:17 -0500 8, 18 -- Received: (qmail 81606 invoked by uid 60001); 14 Aug 2006 11:30:15 -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:Con tent-Transfer-Encoding; 8, 18 -- b=rlmt7902ZNRzFTTFfQF7ruMzaWPunDe6Tu9YxDJ2pD9W3I+BsUwz4JFylLSb1NbV5Y1qFq a1jK6YE5BUUAa9EVhASSCrla6W/fKuGhTPt6o7J7paQwTF6sqlqLCbo9YrFe2LwGUS/eGKK8 kO3I8ZVczxOMyA1VvnaLJ+c9MHHSw= ; 8, 18 -- Message-ID: {20060814113015.81604.qmail-at-web37406.mail.mud.yahoo.com} 8, 18 -- Received: from [80.122.101.102] by web37406.mail.mud.yahoo.com via HTTP; Mon, 14 Aug 2006 04:30:15 PDT 8, 18 -- Date: Mon, 14 Aug 2006 04:30:15 -0700 (PDT) 8, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 18 -- Subject: need infos about e diffraction in TEM 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==============================
==============================Original Headers============================== 23, 26 -- From Wharton.Sinkler-at-uop.com Mon Aug 14 07:37:28 2006 23, 26 -- Received: from AZ18CN849.global.ds.honeywell.com (tmpnat1.honeywell.com [199.64.0.252]) 23, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7ECbPBa021833 23, 26 -- for {microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 07:37:26 -0500 23, 26 -- Received: from dplexch7.uop.com ([138.90.2.112]) by AZ18CN849.global.ds.honeywell.com with Microsoft SMTPSVC(6.0.3790.1830); 23, 26 -- Mon, 14 Aug 2006 05:37:24 -0700 23, 26 -- Received: from DPLEVS1.ad.uop.com ([138.90.2.100]) by dplexch7.uop.com with Microsoft SMTPSVC(6.0.3790.1830); 23, 26 -- Mon, 14 Aug 2006 07:37:23 -0500 23, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 23, 26 -- Content-class: urn:content-classes:message 23, 26 -- MIME-Version: 1.0 23, 26 -- Content-Type: text/plain; 23, 26 -- charset="US-ASCII" 23, 26 -- Subject: RE: [Microscopy] need infos about e diffraction in TEM 23, 26 -- Date: Mon, 14 Aug 2006 07:37:14 -0500 23, 26 -- Message-ID: {62C8A7975D4AE545A23A0CD278BB20A907012FFA-at-DPLEVS1.ad.uop.com} 23, 26 -- X-MS-Has-Attach: 23, 26 -- X-MS-TNEF-Correlator: 23, 26 -- Thread-Topic: [Microscopy] need infos about e diffraction in TEM 23, 26 -- Thread-Index: Aca/ldIkzvC5a3QATaCWBWLUP0Lg8QAA76kw 23, 26 -- From: "Sinkler, Wharton" {Wharton.Sinkler-at-uop.com} 23, 26 -- To: {nizets2-at-yahoo.com} 23, 26 -- Cc: {microscopy-at-microscopy.com} 23, 26 -- X-OriginalArrivalTime: 14 Aug 2006 12:37:23.0010 (UTC) FILETIME=[64000A20:01C6BF9E] 23, 26 -- Content-Transfer-Encoding: 8bit 23, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7ECbPBa021833 ==============================End of - Headers==============================
I am interested to see if someone else from the list chimes in with an alternitive immulination setting specific for histo/path applications, but as far as I know, in terms of training, experiences, and how I teach my students you always set the illumination (condensor lens) for Köhler illumination. This will result in the best resolution.
Steps:
1) Focus sample
2) stop down the field diaphram until you can see it in through the eyepieces (the one closest to the lamp bulb, may not be present on lower cost scopes like general teaching lab scopes. The aperture closest to the condensrr lens is the stage diaphram or aperture diaphram). Note: If the condensor is extremely far from the sample as you indicate move it very close to the sample.
3) Move the condenor lens up and down to focus on the edges of the field diaphram.
4) Spread the field diaphram out to just inside the field of view through the eyepieces. And center it with the condensor centering knobs.
5) Spread the aperture just outside the filed of view.
---- You are now setup for Köhler illumination on that lens, You wil need to repeat this as you change lenses ----
6) For brightfield: Open the Stage Diaphram all the way. While watching through eyepieces close the stage diaphram until you just see a contrast change and stop (If you remove an eyepiece you will note that the stage diaphram will have cropped 20-30% of the filed of view, this is the 70-80% referred to in some texts)
On 12 Aug 2006, at 12:19, gmerkiso-at-iupui.edu wrote:
} } Email: gmerkiso-at-iupui.edu } Name: George Merkison } } Organization: Indiana Blood Centter } } Education: Undergraduate College } } Location: Indianapolis,IN } } Question: When using a light microscope, I was taught to keep the } condenser down for fluids, including urine sediment and cell counts on } hemacytometer, and Nagoette chambers. Is this true and is there a } reference to show other individuals? I can't find a reference. Thanks } for your reply. } } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== 8, 12 -- From } zaluzec-at-microscopy.com Sat Aug 12 12:17:44 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 } k7CHHgWP017031 8, 12 -- for {microscopy-at-microscopy.com} ; Sat, 12 Aug } 2006 12:17:44 -0500 8, 12 -- Mime-Version: 1.0 8, 12 -- X-Sender: } (Unverified) 8, 12 -- Message-Id: } {p06110411c103bd8f2856-at-[206.69.208.22]} 8, 12 -- Date: Sat, 12 Aug } 2006 12:17:41 -0500 8, 12 -- To: microscopy-at-microscopy.com 8, 12 -- } From: gmerkiso-at-iupui.edu (by way of MicroscopyListserver) 8, 12 -- } Subject: AskAMicroscopist: light microscope & fluids 8, 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
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 25, 25 -- From edelmare-at-muohio.edu Mon Aug 14 07:59:55 2006 25, 25 -- Received: from mulnx11.mcs.muohio.edu (mulnx11.mcs.muohio.edu [134.53.6.66]) 25, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7ECxrOR032465 25, 25 -- for {microscopy-at-Microscopy.com} ; Mon, 14 Aug 2006 07:59:54 -0500 25, 25 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 25, 25 -- by mulnx11.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7ECxhgs000409; 25, 25 -- Mon, 14 Aug 2006 08:59:43 -0400 25, 25 -- Received: from emf03 ([134.53.14.97]) 25, 25 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7ECxgqF031494; 25, 25 -- Mon, 14 Aug 2006 08:59:42 -0400 25, 25 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 25, 25 -- To: gmerkiso-at-iupui.edu, microscopy-at-Microscopy.com 25, 25 -- Date: Mon, 14 Aug 2006 09:00:30 -0400 25, 25 -- MIME-Version: 1.0 25, 25 -- Content-type: text/plain; charset=ISO-8859-1 25, 25 -- Subject: Re: [Microscopy] AskAMicroscopist: light microscope & fluids 25, 25 -- Message-ID: {44E03BAE.18044.DB6F204-at-localhost} 25, 25 -- Priority: normal 25, 25 -- In-reply-to: {200608121719.k7CHJ6Ll018387-at-ns.microscopy.com} 25, 25 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 25, 25 -- X-Real-ConnectIP: 134.53.14.97 25, 25 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.66 25, 25 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 25, 25 -- Content-Transfer-Encoding: 8bit 25, 25 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k7ECxrOR032465 ==============================End of - Headers==============================
Nestor J. Zaluzec wrote: } Colleagues... } } As Local Arrangements Co-Chair for the Microscopy & Microanalysis 2006 Meeting } I would like to take a moment on behalf of the LAC, your local host (MMMS) } and the Co-sponsoring societies (MSA, MAS, IMS, MSC/SMC) } to thank you for attending our meeting in Chicago. } } Attendance at the meeting broke all our records for the last decade with } 1873 Scientific registrants and 3173 total attendee's from 33 countries } discussing 938 presentations. Our exhibit featured over 110 companies presenting } the largest state-of-the-art instrumentation exhibit for microscopy and microanalysis } in the world. } } I hope you found the program stimulating, and the venue and ammenties in Chicagoland } to be both exciting and entertaining. The various committee's and meeting personnel } worked hard to make the conference a success and your participation in the symposia } and scientific exhibition was an essential part of this. } } If you have any suggestions and/or comments on any aspect of the meeting, please } feel free to email them to me (zaluzec-at-aaem.amc.anl.gov) and I will insure they } get collected and passed along to future organizers. } } In reminiscence of the meeting, one of the members of our LAC Team (Byran Rabatic) } has created a s video/slide show which superbly captures the spirit of the } MM2006 meeting . If you have a moment you might enjoy } viewing it. Please feel free to download a copy at the following URL: } } http://mm2006.microscopy.org/MM2006large.mov (~ 115 Mb) } or } http://mm2006.microscopy.org/MM2006small.mov ( ~ 28 Mb) } } make sure you have your audio on when you watch it, it is very well done. } } Again, thanks to all of you for attending MM2006 and we hope to see you } next year in Ft. Lauderdale, Florida (http://mm2007.microscopy.org) . } } Cheers.... } } Nestor Zaluzec } Your Friendly Neighborhood LAC Co-Chair } zaluzec-at-aaem.amc.anl.gov } } } ----------------------------------------------------------------------------------- } It's a 106 miles to Chicago. We got a full tank of gas, half a pack of cigarettes, it's dark and we're wearing sunglasses. } ----------------------------------------------------------------------------------- } } }
-- 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, 26 -- From gwe-at-ufl.edu Mon Aug 14 08:57:08 2006 3, 26 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 3, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7EDv5rP011210 3, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 Aug 2006 08:57:07 -0500 3, 26 -- Received: from [10.227.60.63] (empc14419.dhcp.clas.ufl.edu [10.227.60.63]) 3, 26 -- (authenticated bits=0) 3, 26 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k7EDuuqj426042; 3, 26 -- Mon, 14 Aug 2006 09:56:57 -0400 3, 26 -- Message-ID: {44E08128.9070400-at-ufl.edu} 3, 26 -- Date: Mon, 14 Aug 2006 09:56:56 -0400 3, 26 -- From: Greg Erdos {gwe-at-ufl.edu} 3, 26 -- Reply-To: gwe-at-ufl.edu 3, 26 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 3, 26 -- X-Accept-Language: en-us, en 3, 26 -- MIME-Version: 1.0 3, 26 -- To: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} , 3, 26 -- "Microscopy-at-sparc5.microscopy.com" {Microscopy-at-MSA.Microscopy.Com} 3, 26 -- Subject: Re: Thanks for Attending MM2006 3, 26 -- References: {p0611040fc103af90e074-at-[206.69.208.22]} 3, 26 -- In-Reply-To: {p0611040fc103af90e074-at-[206.69.208.22]} 3, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 26 -- Content-Transfer-Encoding: 7bit 3, 26 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 26 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 26 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 3, 26 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
Nice summary, but ... The lamp filament should be centered and in focus in the back focal plane of the objective. This can be done by using the Bertrand lens which places an additional lens in the light path. If you don't have one, an ocular can be removed and the back focal plane examined. A centered pin hole in a tube cap or covering of foil can be used to sharpen the image as can a focusing telescope used in phase contrast.
This is done with the specimen in place and in focus. After focusing and centering the filament, you perform the rest. Unfortunately many modern scope have no provision for this operation.
If you have a rotating stage or objectives which can be centered, life gets a little more complex, but the reward of good illumination is worth it!
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
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edelmare-at-muohio.e du To: frank.karl-at-degussa.com cc: 08/14/2006 09:02 Subject: [Microscopy] Re: AskAMicroscopist: light microscope & fluids AM Please respond to edelmare
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
George:
I am interested to see if someone else from the list chimes in
with an alternitive immulination setting specific for histo/path applications, but as far as I know, in terms of training, experiences, and how I teach my students you always set the illumination (condensor lens) for Köhler illumination. This will result in the best resolution.
Steps:
1) Focus sample
2) stop down the field diaphram until you can see it in through the eyepieces (the one closest to the lamp bulb, may not be present on lower cost scopes like general teaching lab scopes. The aperture closest to the condensrr lens is the stage diaphram or aperture diaphram). Note: If the condensor is extremely far from the sample as you indicate move it very close to the sample.
3) Move the condenor lens up and down to focus on the edges of the field diaphram.
4) Spread the field diaphram out to just inside the field of view through the eyepieces. And center it with the condensor centering knobs.
5) Spread the aperture just outside the filed of view.
---- You are now setup for Köhler illumination on that lens, You wil need to repeat this as you change lenses ----
6) For brightfield: Open the Stage Diaphram all the way. While watching through eyepieces close the stage diaphram until you just see a contrast change and stop (If you remove an eyepiece you will note that the stage diaphram will have cropped 20-30% of the filed of view, this is the 70-80% referred to in some texts)
On 12 Aug 2006, at 12:19, gmerkiso-at-iupui.edu wrote:
} } Email: gmerkiso-at-iupui.edu } Name: George Merkison } } Organization: Indiana Blood Centter } } Education: Undergraduate College } } Location: Indianapolis,IN } } Question: When using a light microscope, I was taught to keep the } condenser down for fluids, including urine sediment and cell counts on } hemacytometer, and Nagoette chambers. Is this true and is there a } reference to show other individuals? I can't find a reference. Thanks } for your reply. } } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== 8, 12 -- From } zaluzec-at-microscopy.com Sat Aug 12 12:17:44 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 } k7CHHgWP017031 8, 12 -- for {microscopy-at-microscopy.com} ; Sat, 12 Aug } 2006 12:17:44 -0500 8, 12 -- Mime-Version: 1.0 8, 12 -- X-Sender: } (Unverified) 8, 12 -- Message-Id: } {p06110411c103bd8f2856-at-[206.69.208.22]} 8, 12 -- Date: Sat, 12 Aug } 2006 12:17:41 -0500 8, 12 -- To: microscopy-at-microscopy.com 8, 12 -- } From: gmerkiso-at-iupui.edu (by way of MicroscopyListserver) 8, 12 -- } Subject: AskAMicroscopist: light microscope & fluids 8, 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
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 25, 25 -- From edelmare-at-muohio.edu Mon Aug 14 07:59:55 2006 25, 25 -- Received: from mulnx11.mcs.muohio.edu (mulnx11.mcs.muohio.edu [134.53.6.66]) 25, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7ECxrOR032465 25, 25 -- for {microscopy-at-Microscopy.com} ; Mon, 14 Aug 2006 07:59:54 -0500 25, 25 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 25, 25 -- by mulnx11.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7ECxhgs000409; 25, 25 -- Mon, 14 Aug 2006 08:59:43 -0400 25, 25 -- Received: from emf03 ([134.53.14.97]) 25, 25 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7ECxgqF031494; 25, 25 -- Mon, 14 Aug 2006 08:59:42 -0400 25, 25 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 25, 25 -- To: gmerkiso-at-iupui.edu, microscopy-at-Microscopy.com 25, 25 -- Date: Mon, 14 Aug 2006 09:00:30 -0400 25, 25 -- MIME-Version: 1.0 25, 25 -- Content-type: text/plain; charset=ISO-8859-1 25, 25 -- Subject: Re: [Microscopy] AskAMicroscopist: light microscope & fluids 25, 25 -- Message-ID: {44E03BAE.18044.DB6F204-at-localhost} 25, 25 -- Priority: normal 25, 25 -- In-reply-to: {200608121719.k7CHJ6Ll018387-at-ns.microscopy.com} 25, 25 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 25, 25 -- X-Real-ConnectIP: 134.53.14.97 25, 25 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.66 25, 25 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 25, 25 -- Content-Transfer-Encoding: 8bit 25, 25 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k7ECxrOR032465 ==============================End of - Headers==============================
==============================Original Headers============================== 51, 20 -- From frank.karl-at-degussa.com Mon Aug 14 09:13:44 2006 51, 20 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 51, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7EEDfZB018378 51, 20 -- for {microscopy-at-msa.microscopy.com} ; Mon, 14 Aug 2006 09:13:42 -0500 51, 20 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [172.20.6.74]) 51, 20 -- by framailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k7EEDRwh014908; 51, 20 -- Mon, 14 Aug 2006 16:13:28 +0200 51, 20 -- In-Reply-To: {200608141302.k7ED25W1002289-at-ns.microscopy.com} 51, 20 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: light microscope & fluids 51, 20 -- To: edelmare-at-muohio.edu, microscopy-at-msa.microscopy.com 51, 20 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 51, 20 -- Message-ID: {OFF522A4D7.949618D6-ON852571CA.004D2386-852571CA.004E1F47-at-degussa.com} 51, 20 -- From: frank.karl-at-degussa.com 51, 20 -- Date: Mon, 14 Aug 2006 10:13:18 -0400 51, 20 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 51, 20 -- 08/14/2006 09:13:31 AM 51, 20 -- MIME-Version: 1.0 51, 20 -- Content-type: text/plain; charset=ISO-8859-1 51, 20 -- Content-Transfer-Encoding: 8bit 51, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7EEDfZB018378 ==============================End of - Headers==============================
I tried to send this in html with my comments in red but the server doesn't allow html so:
REGARDING:
Email: gmerkiso-at-iupui.edu Name: George Merkison Organization: Indiana Blood Centter Education: Undergraduate College Location: Indianapolis,IN
Question: When using a light microscope, I was taught to keep the condenser down for fluids, including urine sediment and cell counts on hemacytometer, and Nagoette chambers. Is this true and is there a reference to show other individuals? I can't find a reference. Thanks for your reply.
MY RESPONSE Are you speaking of a flip out condenser or the whole condenser?
1) Flip-out top piece (a separate lens over the main condenser):
If a flip out - it is useful at low mag (10-20X Obj) because it a) lets the light become more parallel to the axis increasing depth of field, and b) increases contrast, and c) allows the main condenser to stay at the appropriate height (condenser height should allow one to close the field diaphram at the base of the scope and have the edges of this iris clearly in view). However, the gain in contrast (which makes things appear to the eye better - enhances refractve index differences between the crystal or biological specimen and the fluid) through the effective lowering of the numerical aperature (cone of light) will decrease resolution of fine details. Generally one flips out the lens because the gross contrast is more important.
2) Whole condenser:
If it is the whole condenser - the answer is no. The condenser should be set at a particular height to properly converge the light upon the specimen in the correct plane. Lowering the condenser does allow the light to become more parallel to the axis as it penetrates the specimen - which then increases contrast, increases depth of field, but it does not do this as well as a flip out lens. It then places the condenser at the wrong height when switching back to do finer detail work. Also, lowering the whole condenser has a tendency to exaggerate speherical and chromatic aberrations in the lens, allow more scattering of light problems, lower overall light intensity and decrease even-ness (sp?) of background lighting. Not recomended. And certainly don't do it if you have a phase contrast scope as the phase ring and annulus aligment will not match in size and may not be axially aligned either.
3) Bottom Line:
If you don't have a flip-out condenser, just use the iris in the condenser to stop down the cone of light to the specimen (this lowers the effective numerical aperature). You will lose resolution, but gain contrast to see objects easier. You can move it back and forth as necessary.
4) As a side note, with the thickness of some hemacytometers and other chambers, you'll probably not be able to get the condenser close enough (high enough) to the specimen for perfection and lowering the condenser just makes it worse.
Tony
Ps After edelmare-at-muohio.edu's message: Most bioscopes do not allow Kohler illumination, they utilize diffuse illumination which is slightly different and attempts to even the light distribution in the image plane by scattering the light at the field diaphram plane (using ground glass or a diffuser over the light bulb). Kohler uses the plane of the light filament itself projected in the plane of the image. A better form of illumination is to remove the diffuser and place a fiberoptic head at the same location as the light bulb (this is what I do sometimes). It provides cool (thus protects filters) pretty even illumination.
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==============================Original Headers============================== 20, 32 -- From ph2-at-sprynet.com Mon Aug 14 09:39:06 2006 20, 32 -- Received: from elasmtp-dupuy.atl.sa.earthlink.net (elasmtp-dupuy.atl.sa.earthlink.net [209.86.89.62]) 20, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7EEd4Gv032317 20, 32 -- for {Microscopy-at-Microscopy.Com} ; Mon, 14 Aug 2006 09:39:05 -0500 20, 32 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 20, 32 -- s=dk20050327; d=sprynet.com; 20, 32 -- b=hEayTEkAYbd/cD2r7K5BkiGgt7S/RLrkO4HmEl7nkQOimU+NIEZb61tXntJhDIl+; 20, 32 -- h=Received:Reply-To:From:To:Subject:Date:Organization:Message-ID:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Priority:X-MSMail-Priority:X-Mailer:Importance:X-MimeOLE:X-ELNK-Trace:X-Originating-IP; 20, 32 -- Received: from [4.224.108.253] (helo=yourw92p4bhlzg) 20, 32 -- by elasmtp-dupuy.atl.sa.earthlink.net with asmtp (TLSv1:RC4-MD5:128) 20, 32 -- (Exim 4.34) 20, 32 -- id 1GCdaj-0007gj-Vo 20, 32 -- for Microscopy-at-Microscopy.Com; Mon, 14 Aug 2006 10:39:00 -0400 20, 32 -- Reply-To: {ph2-at-sprynet.com} 20, 32 -- From: "Tony Havics" {ph2-at-sprynet.com} 20, 32 -- To: "Microscopy Listserve" {Microscopy-at-Microscopy.Com} 20, 32 -- Subject: RE: [Microscopy] AskAMicroscopist: light microscope & fluids 20, 32 -- Date: Mon, 14 Aug 2006 10:47:01 -0500 20, 32 -- Organization: pH2 20, 32 -- Message-ID: {000601c6bfb8$e4617e00$fd6ce004-at-QEPI.local} 20, 32 -- MIME-Version: 1.0 20, 32 -- Content-Type: text/plain; 20, 32 -- charset="utf-8" 20, 32 -- X-Priority: 3 (Normal) 20, 32 -- X-MSMail-Priority: Normal 20, 32 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 20, 32 -- Importance: Normal 20, 32 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 20, 32 -- X-ELNK-Trace: 6888e50b2be9b4fee5331016acda17f94d0058a98e3ac958cb4740967cf4ba72350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c 20, 32 -- X-Originating-IP: 4.224.108.253 20, 32 -- Content-Transfer-Encoding: 8bit 20, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7EEd4Gv032317 ==============================End of - Headers==============================
"CCD Cameras for Ultra Low Light Microscopy: Technology Overview and Issues to Consider Before Buying."
This Friday (18-August) at 11:30 AM (New York time). See details below.
Admission is free, but connection lines are limited so reserve yours now.
-------------------------------------------------------------- TO RESERVE YOUR CONNECTION LINE -------------------------------------------------------------- Click this URL: https://premconf.webex.com/premconf/j.php?ED=87414217&RG=1
Click REGISTER and complete the requested information. You will be sent a link that gives you access to Friday's meeting.
Details: Cameras utilizing Intensification and Electron-Multiplication technologies are used by life and physical science microscopists to enable the imaging of dynamic events under the most challenging low-light conditions. QImaging is presenting a free, live, interactive, web-based seminar on these two signal amplification technologies on Friday (18-August) at 11:30 AM (New York time).
Who should attend: Anyone interested in learning how these technologies work, what their fundamental limitations are, and what should be considered before acquiring such a camera for their lab should attend. Details are below. There is no cost, but connection lines are limited so reserve yours now.
-------------------------------------------------------------- TO RESERVE YOUR CONNECTION LINE -------------------------------------------------------------- Click this URL: https://premconf.webex.com/premconf/j.php?ED=87414217&RG=1
Click REGISTER and complete the requested information. You will be sent a link that gives you access to Friday's meeting.
---------------------------------------- MEETING SUMMARY ---------------------------------------- Name: "CCD Cameras for Ultra Low Light Microscopy: Technology Overview and Issues to Consider Before Buying."
Hi Everyone, Can anyone recommend a good, easy to use, software package for creating reconstructions from serial TEM sections that will run a PC platform? Would prefer public domain but am open to inexpensive commercial options as well.
Please reply directly and not to the listserve.
Thanks John Shields EM Lab UGA Athens, GA jpshield-at-uga.edu John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602
706-542-4080
==============================Original Headers============================== 4, 21 -- From jpshield-at-uga.edu Mon Aug 14 14:21:15 2006 4, 21 -- Received: from puntd3.cc.uga.edu (puntd3.cc.uga.edu [128.192.1.109]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7EJLCK8024969 4, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 Aug 2006 14:21:14 -0500 4, 21 -- Received: from punts4.cc.uga.edu (punts4.cc.uga.edu [128.192.1.115]) 4, 21 -- by puntd3.cc.uga.edu (MOS 3.7.4b-GA) 4, 21 -- with ESMTP id ABJ51305; 4, 21 -- Mon, 14 Aug 2006 15:21:10 -0400 (EDT) 4, 21 -- Received: (from punts4.cc.uga.edu [128.192.63.198]) 4, 21 -- by punts4.cc.uga.edu (MOS 3.7.4c) 4, 21 -- with HTTPS/1.1 id BCQ04218 (AUTH jpshield-at-uga.edu); 4, 21 -- Mon, 14 Aug 2006 15:20:29 -0400 (EDT) 4, 21 -- From: John Shields {jpshield-at-uga.edu} 4, 21 -- Subject: TEM serial section 3D 4, 21 -- To: msa listserve {Microscopy-at-MSA.Microscopy.Com} 4, 21 -- X-Mailer: Mirapoint Webmail Direct 3.7.4c 4, 21 -- MIME-Version: 1.0 4, 21 -- Content-Type: text/plain; charset=us-ascii 4, 21 -- Content-Transfer-Encoding: 7bit 4, 21 -- Message-Id: {20060814152029.BCQ04218-at-punts4.cc.uga.edu} 4, 21 -- Date: Mon, 14 Aug 2006 15:20:29 -0400 (EDT) ==============================End of - Headers==============================
Anyone have a working LKB Bromma 2208 multiplate wax heater/hot plate warmer for donation or (cheap) sale? Thanks.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Mon Aug 14 14:33:51 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 k7EJXnLC029006 3, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 14:33:50 -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 k7EK7IqG006280 3, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 16:07:18 -0400 3, 20 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 20 -- Mon, 14 Aug 2006 15:33:44 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f06230917c106804d8e23-at-[141.209.160.249]} 3, 20 -- Date: Mon, 14 Aug 2006 15:33:41 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: LKB 2208 wax heater 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 14 Aug 2006 19:33:44.0946 (UTC) FILETIME=[8E64F520:01C6BFD8] 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==============================
Dear Listserver: We have a very small electron microscopy laboratory with 1970;s state of the art equipment (Philips 201 and ETEC Autoscan). I am concerned about the dwindling shortage and increased costs of film, chemistry and paper. I would like to know if either or both of these instruments could be easily converted to digial from film recording. I have not kept up with the ccd camera prices and someone told me they wondered if a good digital camera with an adapter would be available. What kind of reasonable options are there for the old but very functional TEM and SEM? Thanks for your advice. Barbara Plowman
Barbara L. Plowman University of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster St. Rm 636B San Francisco, CA 94115 ph: 415-929-6692 email: Bplowman-at-pacific.edu
==============================Original Headers============================== 3, 16 -- From Bplowman-at-Pacific.edu Mon Aug 14 14:55:18 2006 3, 16 -- Received: from sfmail.dental.pacific.edu (sfmail.dental.uop.edu [138.9.130.8]) 3, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7EJtG5I013380 3, 16 -- for {Microscopy-at-microscopy.org} ; Mon, 14 Aug 2006 14:55:17 -0500 3, 16 -- Received: from SFUOP-MTA by sfmail.dental.pacific.edu 3, 16 -- with Novell_GroupWise; Mon, 14 Aug 2006 12:55:12 -0700 3, 16 -- Message-Id: {44E072A602000049000042E7-at-sfmail.dental.pacific.edu} 3, 16 -- X-Mailer: Novell GroupWise Internet Agent 7.0 3, 16 -- Date: Mon, 14 Aug 2006 12:55:02 -0700 3, 16 -- From: "Barbara Plowman" {Bplowman-at-Pacific.edu} 3, 16 -- To: {Microscopy-at-microscopy.org} 3, 16 -- Subject: Converting to digital 3, 16 -- Mime-Version: 1.0 3, 16 -- Content-Type: text/plain; charset=US-ASCII 3, 16 -- Content-Transfer-Encoding: 7bit 3, 16 -- Content-Disposition: inline ==============================End of - Headers==============================
Hi We do hundreds of 3D reconstruction from TEM. Just to make a stack of images I use QuickTime. That way people can 'scrub' through the data to see the TEMs. There is no package available (that I know of) that will take raw TEM data and output a 3D rendering that is useful. I use Volocity (not public domain, nor cheep) to do the 3D reconstructions and analytical work. Unless others have ideas, I think you will have to be doing some work with the raw data before the 3D work can be done. Feel free to contact me in you have any questions. If you are only doing a small number, I could do it for you. 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 Aug 14, 2006, at 12:37 PM, jpshield-at-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 Everyone, } Can anyone recommend a good, easy to use, software package for } creating reconstructions from serial TEM sections that will run a } PC platform? } Would prefer public domain but am open to inexpensive commercial } options as well. } } Please reply directly and not to the listserve. } } Thanks } John Shields } EM Lab } UGA Athens, GA } jpshield-at-uga.edu } John P. Shields } Center for Ultrastructural Research } 151 Barrow Hall } University of Georgia } Athens, GA 30602 } } 706-542-4080 } } ==============================Original } Headers============================== } 4, 21 -- From jpshield-at-uga.edu Mon Aug 14 14:21:15 2006 } 4, 21 -- Received: from puntd3.cc.uga.edu (puntd3.cc.uga.edu } [128.192.1.109]) } 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k7EJLCK8024969 } 4, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 Aug 2006 } 14:21:14 -0500 } 4, 21 -- Received: from punts4.cc.uga.edu (punts4.cc.uga.edu } [128.192.1.115]) } 4, 21 -- by puntd3.cc.uga.edu (MOS 3.7.4b-GA) } 4, 21 -- with ESMTP id ABJ51305; } 4, 21 -- Mon, 14 Aug 2006 15:21:10 -0400 (EDT) } 4, 21 -- Received: (from punts4.cc.uga.edu [128.192.63.198]) } 4, 21 -- by punts4.cc.uga.edu (MOS 3.7.4c) } 4, 21 -- with HTTPS/1.1 id BCQ04218 (AUTH jpshield-at-uga.edu); } 4, 21 -- Mon, 14 Aug 2006 15:20:29 -0400 (EDT) } 4, 21 -- From: John Shields {jpshield-at-uga.edu} } 4, 21 -- Subject: TEM serial section 3D } 4, 21 -- To: msa listserve {Microscopy-at-MSA.Microscopy.Com} } 4, 21 -- X-Mailer: Mirapoint Webmail Direct 3.7.4c } 4, 21 -- MIME-Version: 1.0 } 4, 21 -- Content-Type: text/plain; charset=us-ascii } 4, 21 -- Content-Transfer-Encoding: 7bit } 4, 21 -- Message-Id: {20060814152029.BCQ04218-at-punts4.cc.uga.edu} } 4, 21 -- Date: Mon, 14 Aug 2006 15:20:29 -0400 (EDT) } ==============================End of - } Headers==============================
==============================Original Headers============================== 10, 22 -- From Elliott-at-arizona.edu Mon Aug 14 15:03:35 2006 10, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 10, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7EK3Xi0022963 10, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 Aug 2006 15:03:34 -0500 10, 22 -- Received: from localhost (gimli.email.arizona.edu [10.0.0.223]) 10, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 5B24BF1986A 10, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 Aug 2006 13:03:27 -0700 (MST) 10, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 10, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 9E47FF1888E 10, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 14 Aug 2006 12:55:15 -0700 (MST) 10, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 10, 22 -- In-Reply-To: {200608141937.k7EJbJ9U003496-at-ns.microscopy.com} 10, 22 -- References: {200608141937.k7EJbJ9U003496-at-ns.microscopy.com} 10, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 10, 22 -- Message-Id: {39C9C0E8-E1B9-4E04-BD57-06538D9199BF-at-arizona.edu} 10, 22 -- Content-Transfer-Encoding: 7bit 10, 22 -- From: David Elliott {Elliott-at-arizona.edu} 10, 22 -- Subject: Re: [Microscopy] TEM serial section 3D 10, 22 -- Date: Mon, 14 Aug 2006 12:55:14 -0700 10, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 10, 22 -- X-Mailer: Apple Mail (2.752.2) 10, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Dear Listserver: We have a very small electron microscopy laboratory with 1970;s state of the art equipment (Philips 201 and ETEC Autoscan). I am concerned about the dwindling shortage and increased costs of film, chemistry and paper. I would like to know if either or both of these instruments could be easily converted to digial from film recording. I have not kept up with the ccd camera prices and someone told me they wondered if a good digital camera with an adapter would be available. What kind of reasonable options are there for the old but very functional TEM and SEM? Thanks for your advice. Barbara Plowman
Barbara L. Plowman University of the Pacific Arthur A. Dugoni School of Dentistry 2155 Webster St. Rm 636B San Francisco, CA 94115 ph: 415-929-6692 email: Bplowman-at-pacific.edu
==============================Original Headers============================== 3, 16 -- From Bplowman-at-Pacific.edu Mon Aug 14 15:14:40 2006 3, 16 -- Received: from sfmail.dental.pacific.edu (sfmail.dental.uop.edu [138.9.130.8]) 3, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7EKEcV2001664 3, 16 -- for {Microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 15:14:39 -0500 3, 16 -- Received: from SFUOP-MTA by sfmail.dental.pacific.edu 3, 16 -- with Novell_GroupWise; Mon, 14 Aug 2006 13:14:37 -0700 3, 16 -- Message-Id: {44E0773302000049000042F2-at-sfmail.dental.pacific.edu} 3, 16 -- X-Mailer: Novell GroupWise Internet Agent 7.0 3, 16 -- Date: Mon, 14 Aug 2006 13:14:27 -0700 3, 16 -- From: "Barbara Plowman" {Bplowman-at-Pacific.edu} 3, 16 -- To: {Microscopy-at-microscopy.com} 3, 16 -- Subject: Converting to digital 3, 16 -- Mime-Version: 1.0 3, 16 -- Content-Type: text/plain; charset=US-ASCII 3, 16 -- Content-Transfer-Encoding: 7bit 3, 16 -- Content-Disposition: inline ==============================End of - Headers==============================
We prepare thin sections of epoxy embedded samples and came across an LKB 2188 microtome. We currently cut these with a wafering blade with great difficulty.
Would anyone be familiar with this unit? Can we get blades to section samples upwards of approximately 1/2 inch in diameter?
Thanks for any help.
Alan Stone
==============================Original Headers============================== 7, 19 -- From as-at-astonmet.com Mon Aug 14 20:19:24 2006 7, 19 -- Received: from outbound1.mail.tds.net (outbound1.mail.tds.net [216.170.230.91]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7F1JO14016683 7, 19 -- for {microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 20:19:24 -0500 7, 19 -- Received: from outaamta01.mail.tds.net (outaamta01.mail.tds.net [216.170.230.31]) 7, 19 -- by outbound1.mail.tds.net (8.13.6/8.13.4) with ESMTP id k7F1JNWU027950 7, 19 -- for {microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 20:19:23 -0500 7, 19 -- Received: from D95S7761.astonmet.com ([216.170.179.105]) 7, 19 -- by outaamta01.mail.tds.net with ESMTP 7, 19 -- id {20060815011923.INNR5875.outaamta01.mail.tds.net-at-D95S7761.astonmet.com} 7, 19 -- for {microscopy-at-microscopy.com} ; Mon, 14 Aug 2006 20:19:23 -0500 7, 19 -- Message-Id: {6.2.0.14.2.20060814195433.020a5fb8-at-pop.tds.net} 7, 19 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.0.14 7, 19 -- Date: Mon, 14 Aug 2006 20:19:23 -0500 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- From: Alan Stone {as-at-astonmet.com} 7, 19 -- Subject: lkb 2188 microtome information 7, 19 -- Mime-Version: 1.0 7, 19 -- 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 alerch-at-mcw.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: alerch-at-mcw.edu Name: Alexandra Lerch-Gaggl
Question: We have a Leica TCS SP2 with a 568nm laser in our facility. We had the tube remanufactured recently and I find now that over the time of an experiment the laser output decreases significantly, so that the intense signal i had at the beginning of the session is so deminished that I completely loose the image. When I open the box, which houses the laser, the power supply and the ventilator (remark on the side: I am allowed to do that, since I direct the facility and take care of our equipment), I see the signal return to its original intensity in a matter of a minute. My question is now: Is this related to overheating due to unsufficient ventilation or ... what else? I would appreciate if you could share your ideas/solutions with me. Thank you.
Barbara L. Plowman wrote: ===================================================== We have a very small electron microscopy laboratory with 1970;s state of the art equipment (Philips 201 and ETEC Autoscan). I am concerned about the dwindling shortage and increased costs of film, chemistry and paper. I would like to know if either or both of these instruments could be easily converted to digial from film recording. I have not kept up with the ccd camera prices and someone told me they wondered if a good digital camera with an adapter would be available. What kind of reasonable options are there for the old but very functional TEM and SEM? Thanks for your advice. =====================================================
These are really two different issues. For the Philips 201 TEM, there are cameras available from several different manufacturers. For example, see URL http://www.sia-cam.com/ There is also the possibility of digial imaging plates. http://www.ditabis.de/
For the ETEC SEM, you can convert from analog to digital with a number of different software/hardware products, one being the Orion Frame Grabber System for SEMs. See URL http://www.2spi.com/catalog/instruments/ORION_Digital_Image_Acquisition_System.html
As is the case with most products in EM, there is a wide range of product pricing, depending on budget and application.
It is off the topic but these two instruments are excellent for teaching purposes. When a student learns how to master the operation of either, they really do know how a TEM or SEM really "works". There is no software to do the thinking for you........
Chuck
Disclaimer: SPI Supplies offers the Orion Frame Grabber System for SEMs via the website indicated above.
=================================================== 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, 26 -- From cgarber-at-2spi.com Mon Aug 14 21:33:50 2006 12, 26 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 12, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7F2XnDL005870 12, 26 -- for {microscopy-at-msa.microscopy.com} ; Mon, 14 Aug 2006 21:33:49 -0500 12, 26 -- Received: from yourb27fb1c401 (c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118]) 12, 26 -- (authenticated bits=0) 12, 26 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id k7F2XfxV000987 12, 26 -- for {microscopy-at-msa.microscopy.com} ; Mon, 14 Aug 2006 22:33:49 -0400 12, 26 -- X-IDV-FirstRcvd: c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118] 12, 26 -- X-IDV-HELO: yourb27fb1c401 12, 26 -- X-IDV-Authenticated-User: cgarber 12, 26 -- Message-ID: {026e01c6c013$3af93500$6401a8c0-at-yourb27fb1c401} 12, 26 -- From: "Garber, Charles A" {cgarber-at-2spi.com} 12, 26 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 12, 26 -- Subject: converting to digital 12, 26 -- Date: Mon, 14 Aug 2006 22:33:44 -0400 12, 26 -- MIME-Version: 1.0 12, 26 -- Content-Type: text/plain; 12, 26 -- format=flowed; 12, 26 -- charset="iso-8859-1"; 12, 26 -- reply-type=original 12, 26 -- Content-Transfer-Encoding: 7bit 12, 26 -- X-Priority: 3 12, 26 -- X-MSMail-Priority: Normal 12, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 12, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
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Question: I am using Aurion protein-A conjugated to 10 nm gold beads to detect viral antigens in an infected cell line - ultrathin sectioned and immnunostained. I am seeing a capping at the poles of cell sections and am wondering if this can be due to the beads and/or to the presence of serum blocking antibodies. The EMS Immnunogold Product Newsletter talks of Ultra-small reagent use for "No capping or patching, each probe unit consists of a single ligand". By the way, I am not seeing capping with IgM conjugated to 10nm beads and am unsure at this time about IgG conjugated beads.
On Aug 14, 2006, at 4:30 AM, nizets2-at-yahoo.com wrote:
} 1) Can I use e diffraction to distinguish both types } of particles? (to verify the purity of the powder) } 2) What kind of information about the cristal } structure can e diffraction give me in this case? } 3) Can I detect a change -and which change- to the } cristal structure using this technique if the mineral } is heat and treated with strong acids? (which actually } modifies the structure) } Dear Stephane, Just an addition to Wharton's excellent response. If your specimen consists of crystals too small to isolate one or two, you will likely have a ring pattern--either a solid ring if the selected area contains very many crystals or a series of spots arranged on a ring if there are fewer crystals. Measuring the diameter of the ring and comparing it to that for a known substance will give you spacings, which you can fit to those in model structures, and changes in the relative intensities of the rings can give structural information also (This could be due to slight atomic displacements that do not affect the unit cell, but change the relationships of the scattering from the atoms in the crystal.) One of the recent publications from our lab (Wright, E. R., Iancu, C. V., Tivol, W. F., and Jensen, G. J., Observations on the Behavior of Vitreous Ice at ~82 and ~12 K. Journal of Structural Biology, (2006) in press.) used ED to determine the dose at which low density amorphous ice underwent a transition to high density amorphous ice, so you could expect to see similar changes. 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 Aug 15 15:55:22 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 k7FKtMhB012610 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 15 Aug 2006 15:55:22 -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 177C3109AFE 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 15 Aug 2006 13:55:22 -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 4261A109B31 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 15 Aug 2006 13:55:16 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200608141130.k7EBUZCV010760-at-ns.microscopy.com} 4, 22 -- References: {200608141130.k7EBUZCV010760-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {5f33891af715f7b0adde84f0aae90373-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] need infos about e diffraction in TEM 4, 22 -- Date: Tue, 15 Aug 2006 13:57:39 -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==============================
I have been imaging negatively stained virions and isolated viral proteins and was wondering if anyone has ever applied blind deconvolution to images of negatively stained samples? Are there any references or caveats? Thanks!
George
George P. Leser, PhD Dept. Biochemistry, Molecular Biology, and Cell Biology Northwestern University Hogan 2-100 2153 North Campus Drive Evanston, IL 60208
g-leser-at-northwestern.edu
==============================Original Headers============================== 8, 22 -- From g-leser-at-northwestern.edu Tue Aug 15 16:25:56 2006 8, 22 -- Received: from merle.it.northwestern.edu (merle.it.northwestern.edu [129.105.16.57]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7FLPuQS023253 8, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 15 Aug 2006 16:25:56 -0500 8, 22 -- Received: from hal90003ecc03cf (dhcp-129-105-38-214.bmbcb.northwestern.edu [129.105.38.214]) 8, 22 -- by merle.it.northwestern.edu (Postfix) with SMTP id 447142F0B0 8, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 15 Aug 2006 16:25:56 -0500 (CDT) 8, 22 -- Message-ID: {003001c6c0b1$694c72c0$d6266981-at-hal90003ecc03cf} 8, 22 -- From: "George P. Leser" {g-leser-at-northwestern.edu} 8, 22 -- To: "1A-Microscopy Listserv" {Microscopy-at-microscopy.com} 8, 22 -- Subject: deconvolution of negatively stained EM images 8, 22 -- Date: Tue, 15 Aug 2006 16:26:02 -0500 8, 22 -- MIME-Version: 1.0 8, 22 -- Content-Type: text/plain; 8, 22 -- format=flowed; 8, 22 -- charset="iso-8859-1"; 8, 22 -- reply-type=original 8, 22 -- Content-Transfer-Encoding: 7bit 8, 22 -- X-Priority: 3 8, 22 -- X-MSMail-Priority: Normal 8, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 8, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jchampagne-at-caseforensicscorp.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, August 15, 2006 at 14:06:51 ---------------------------------------------------------------------------
Email: jchampagne-at-caseforensicscorp.com Name: Joseph Champagne
Organization: CASE Forensics - Used Microscopes
Education: 9-12th Grade High School
Location: Leavenworth, WA
Question: I would like some advise and/or recommendations on purchasing a good used microscope. What manufacturer(s), magnification power, light capability, where to buy, etc? Mono vs. Stereo?
I want to use it with my high school age children to look at plant and animal cells, and disected samples.
In theory, any conjugate which has more than one binding site, could induce capping or patching. This is true for all the conventional immuno-reagents, i.e. the big particles that likely have more than one specific binding molecule adsorbed or bound to the particle's surface. Ultra small particle based reagents, as quoted from the EMS Newsletter, are of a different built: they are proteins bound with one (or more) ultra small particles as opposed to a 10nm particle which is coated with more than one protein molecule. If you used serum in your blocking/incubation solutions the protein A gold reagent may have reacted with available immunoglobulins, which in the end may be causing the phenomenon as well.
Capping and patching may also be induced by the primary antibody, especially when more than 1 epitope is involved.
With more info I may be able to give you a better idea. Please contact me off-list.
Jan Leunissen
Aurion - President Electron Microscopy Research Advisor Costerweg 5 Dept Anatomy and Structural Biology 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4795465 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz ------------------------------------------------------------------------ --------
==============================Original Headers============================== 13, 20 -- From leunissen-at-aurion.nl Tue Aug 15 20:44:47 2006 13, 20 -- Received: from fep04.xtra.co.nz (fep04.xtra.co.nz [210.54.141.242]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7G1ikZi015313 13, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 15 Aug 2006 20:44:46 -0500 13, 20 -- Received: from [192.168.1.50] (really [222.152.164.98]) by fep04.xtra.co.nz 13, 20 -- with ESMTP 13, 20 -- id {20060816014436.XEKQ19789.fep04.xtra.co.nz-at-[192.168.1.50]} ; 13, 20 -- Wed, 16 Aug 2006 13:44:36 +1200 13, 20 -- In-Reply-To: {200608160321.AA262209640-at-aurion.nl} 13, 20 -- References: {200608160321.AA262209640-at-aurion.nl} 13, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 13, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 13, 20 -- Message-Id: {262522CB-EC3C-4D2B-98E4-C6866BB65965-at-aurion.nl} 13, 20 -- Cc: Microscopy-at-microscopy.com 13, 20 -- Content-Transfer-Encoding: 7bit 13, 20 -- From: Jan Leunissen {leunissen-at-aurion.nl} 13, 20 -- Subject: Re: [Microscopy] viaWWW: Capping or Patching 13, 20 -- Date: Wed, 16 Aug 2006 13:44:35 +1200 13, 20 -- To: kmoulton-at-usm.maine.edu 13, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
Hi I need help in processing lymph nodes for TEM. I need a specific protocol for Lymph Nodes. The protocol that I'm using hasn't worked for this tissue. This protocol works with other tissues such as skin and heart 1- chemical fixation (glutaraldehyde + formaldehyde + tannic acid in cacodylate buffer for one hour) 2- secondary fixation with osmium 3- uranyl acetate overnight 4- dehydrations with increasing concentrations of ethanol (20 minutes each step) 5- resin infiltration with increasing concentration of resin (1 hour each step) 6- overnight incubation with 100% resin 7- next day 1 hour incubation with 100% resin 8- polymerization of the blocks containing the samples for 2 days at 60 degrees celsius.
Thanks!
Jaime
Jaime Llodra Graduate Student Sackler Institute NYU School of Medicine NY, NY 10016
==============================Original Headers============================== 4, 23 -- From jal490-at-nyu.edu Tue Aug 15 20:51:02 2006 4, 23 -- Received: from mx4.nyu.edu (MX4.NYU.EDU [128.122.108.244]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7G1p15k025545 4, 23 -- for {Microscopy-at-Microscopy.com} ; Tue, 15 Aug 2006 20:51:02 -0500 4, 23 -- Received: from mail.nyu.edu (H3.HOME.NYU.EDU [128.122.108.43]) 4, 23 -- by mx4.nyu.edu (8.13.6/8.13.6) with ESMTP id k7G1p1a6028380 4, 23 -- for {Microscopy-at-Microscopy.com} ; Tue, 15 Aug 2006 21:51:01 -0400 (EDT) 4, 23 -- Received: from [216.165.126.103] by mail.alt.home.nyu.edu (mshttpd); Tue, 4, 23 -- 15 Aug 2006 21:51:01 -0400 4, 23 -- From: Jaime A Llodra {jal490-at-nyu.edu} 4, 23 -- To: Microscopy-at-Microscopy.com 4, 23 -- Message-ID: {d55d8f9f32fb.44e241c5-at-mail.nyu.edu} 4, 23 -- Date: Tue, 15 Aug 2006 21:51:01 -0400 4, 23 -- X-Mailer: Sun Java(tm) System Messenger Express 6.2-4.03 (built Sep 22 4, 23 -- 2005) 4, 23 -- MIME-Version: 1.0 4, 23 -- Content-Language: en 4, 23 -- Subject: TEM - lymph node processing 4, 23 -- X-Accept-Language: en 4, 23 -- Priority: normal 4, 23 -- Content-Type: text/plain; charset=us-ascii 4, 23 -- Content-Disposition: inline 4, 23 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Hi, I would appreciate the advice on the thioflavin-S staining. I have two precious sections which were previously studied for presence of inorganic deposits. The 40 micron thick sections from 4% PFA fixed brain tissue were mounted on slide, air-dried, and evaluated by x-ray spectroscopy. Now I would like to demonstrate amyloid but do not want to take any chances with the very important sample. I am looking for the best protocol to use on the sections ?abused? by air-drying. Thank you for your consideration, Albina
-- MIKHAYLOVA,ALBINA, PhD Post Doctoral Research Associate Materials Science and Engineering University of Florida
==============================Original Headers============================== 4, 21 -- From amich-at-ufl.edu Tue Aug 15 21:45:27 2006 4, 21 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7G2jQJ7003922 4, 21 -- for {Microscopy-at-Microscopy.com} ; Tue, 15 Aug 2006 21:45:27 -0500 4, 21 -- Received: from osgjas01.cns.ufl.edu (osgjas01.cns.ufl.edu [128.227.74.131]) 4, 21 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k7G2jMKZ1974384 4, 21 -- for {Microscopy-at-Microscopy.com} ; Tue, 15 Aug 2006 22:45:22 -0400 4, 21 -- Message-ID: {1121615292.49521155696322076.JavaMail.osg-at-osgjas01.cns.ufl.edu} 4, 21 -- Date: Tue, 15 Aug 2006 22:45:22 -0400 (EDT) 4, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 4, 21 -- To: Microscopy-at-Microscopy.com 4, 21 -- Subject: thioflavin-S 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: 68.220.153.163 [68.220.153.163] 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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I'll be happy to provide specific advice, but I need to know more about your needs & experience. I'm reluctant to recommend a used scope unless you have enough experience to recognize problems, and to fix them. Does "plant and animal cells" mean purchased, prepared slides? And what do you mean by "dissected samples"? What is your budget?
Although the microscope-buying advice on the MICRO website (URL below) is intended for a younger age group, you may find it a useful place to begin. There are several books in the MICRO bibliography that can help; again, I need to know more to make suggestions.
Caroline
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 5, 19 -- From schooley-at-mcn.org Tue Aug 15 23:50:47 2006 5, 19 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7G4olg8016555 5, 19 -- for {microscopy-at-microscopy.com} ; Tue, 15 Aug 2006 23:50:47 -0500 5, 19 -- Received: from [66.52.139.254] (helo=[10.0.1.2]) 5, 19 -- by dns4.mcn.org with esmtpa (Exim 4.60) 5, 19 -- (envelope-from {schooley-at-mcn.org} ) 5, 19 -- id J42QSJ-000HO6-4P; Tue, 15 Aug 2006 21:50:44 -0700 5, 19 -- Mime-Version: 1.0 5, 19 -- Message-Id: {a06200700c1084e7f6931-at-[10.0.1.2]} 5, 19 -- In-Reply-To: {200608160122.k7G1MtaK013462-at-ns.microscopy.com} 5, 19 -- References: {200608160122.k7G1MtaK013462-at-ns.microscopy.com} 5, 19 -- Date: Tue, 15 Aug 2006 21:44:17 -0700 5, 19 -- To: jchampagne-at-caseforensicscorp.com 5, 19 -- From: Caroline Schooley {schooley-at-mcn.org} 5, 19 -- Subject: Re: [Microscopy] [Filtered] AskAMicroscopist: Used Microscopes 5, 19 -- with my high school 5, 19 -- Cc: microscopy-at-microscopy.com 5, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
At 20:54 15/08/2006 -0500, you wrote: hi Jaime, If you were more specific it would be easier to help.
In what way has your current protocol not worked, e.g. did the blocks not section well? Is the ultrastructural preservation poor? Was there a lot of deposit on the sections?
I presume you are using epoxy resin processing, in which case I suggest the following:
Make sure you are working with tissue of the correct dimensions, particularly the thickness which should not exceed 1mm. Next I would leave out the tanic acid and formaldehyde and fix in fresh ultrastructural grade gluteraldehyde (2 - 2.5% in 0.1 M phosphtae or cac buffer) overnight at 4'C. Buffer rinse and secondary fix in 1% OsO4. Cut way down on the U/A time to about 30mins in 1% aqueous. Your dehydrations times are OK if you are using thin slices of node but make sure the final dehydrant is dry by keeping your ethanol or acetone over a molecular seive. If you are using ethanol you must follow this with a link reagent such as proylene oxide and use this to mix with the resin for your infiltrations. If using acetone you dont need a link reagent as you can dilute your resin infiltration solns with acetone. If you are having sectioning problems, a bit of heat (but don't exceed 30 mins at 60'C) plus vacuum helps with the infiltration.
Hope this helps, but get back to me with specifics if you need any more guidance.
Alastair
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Alastair McKinnon Histology & EM Facility Manager University of Aberdeen, Institute of Medical Science Foresterhill, Aberdeen, AB25 2ZD tel: +44(0) 1224 552923 (office); +44(0) 1224 555911 (lab) fax: +44(0) 1224 559533; email: a.d.mckinnon-at-abdn.ac.uk www.abdn.ac.uk/ims/h-em
==============================Original Headers============================== 12, 18 -- From a.d.mckinnon-at-abdn.ac.uk Wed Aug 16 03:01:35 2006 12, 18 -- Received: from mailhub1.abdn.ac.uk (mailhub1.abdn.ac.uk [139.133.7.28]) 12, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7G81Z9k029940 12, 18 -- for {Microscopy-at-Microscopy.Com} ; Wed, 16 Aug 2006 03:01:35 -0500 12, 18 -- Received: from med-0069.ims.abdn.ac.uk ([139.133.159.90] helo=med-0069.abdn.ac.uk) 12, 18 -- by mailhub1.abdn.ac.uk with esmtp (Exim 4.52) 12, 18 -- id 1GDGLG-0000fE-EN; Wed, 16 Aug 2006 09:01:34 +0100 12, 18 -- Message-Id: {5.2.1.1.0.20060816084017.011188a8-at-mailms.abdn.ac.uk} 12, 18 -- X-Sender: pat081-at-mailms.abdn.ac.uk 12, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 12, 18 -- Date: Wed, 16 Aug 2006 09:01:35 +0100 12, 18 -- To: jal490-at-nyu.edu 12, 18 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 12, 18 -- Subject: Re: [Microscopy] TEM - lymph node processing 12, 18 -- Cc: Microscopy-at-Microscopy.Com 12, 18 -- In-Reply-To: {200608160154.k7G1sWKD002024-at-ns.microscopy.com} 12, 18 -- Mime-Version: 1.0 12, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Thanks for the tip, but this book is not available at Amazon. I did a search on the www but I cannot find a distributor (it is probably too old).
I already spent one day on the link given by Sousan (very nice).
Would there be another good reference book (more recent perhaps) which does the same job as Hirsch's?
Regards,
Stephane
--- Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} wrote:
} Stephane } } Purchase a copy of Hirsch's book. } Go to Amazon.com. } Search for "transmission electron hirsch". } } regards, } } Jim } } } From mail-at-ns.microscopy.com Mon Aug 14 07:30:49 } 2006 } } Date: Mon, 14 Aug 2006 06:33:44 -0500 } } To: jquinn-at-www.matscieng.sunysb.edu } } From: nizets2-at-yahoo.com } } Reply-to: nizets2-at-yahoo.com } } X-Resent-From: "Microscopy Listserver" } {microscopy-at-microscopy.com} } } Subject: [Microscopy] need infos about e } diffraction in TEM } } Errors-To: } MicroscopyListSpamFilter-at-microscopy.com } } X-lewp: MicroscopyListSpam NAGS } } } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } ---------------------------------------------------------------------------- } } } } Dear listers, } } } } Although I understand the principle behind } electron } } diffraction in TEM I have no idea about the kind } of } } informations this technique gives and how to } interpret } } the diffraction pattern. } } If you could give me a www address which explains } that } } I would be much grateful. } } } } Now a very practical problem could be perhaps } used as } } an example: we have a mixture of aluminosilicate } } mineral (I have cut 70 nm sections) with approx. } 90% } } of mordenite and 10% quarz. } } 1) Can I use e diffraction to distinguish both } types } } of particles? (to verify the purity of the } powder) } } 2) What kind of information about the cristal } } structure can e diffraction give me in this case? } } 3) Can I detect a change -and which change- to } the } } cristal structure using this technique if the } mineral } } is heat and treated with strong acids? (which } actually } } modifies the structure) } } } } We have also a tilt stage for tomography. Can } this } } bring further informations? } } } } Regards, } } } } Stephane } } } } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? Yahoo! Mail has the best spam } protection around } } http://mail.yahoo.com } } } } ==============================Original } Headers============================== } } 8, 18 -- From nizets2-at-yahoo.com Mon Aug 14 } 06:30:17 2006 } } 8, 18 -- Received: from } web37406.mail.mud.yahoo.com } (web37406.mail.mud.yahoo.com [209.191.91.138]) } } 8, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } k7EBUGao010591 } } 8, 18 -- for {microscopy-at-microscopy.com} ; Mon, } 14 Aug 2006 06:30:17 -0500 } } 8, 18 -- Received: (qmail 81606 invoked by uid } 60001); 14 Aug 2006 11:30:15 -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=rlmt7902ZNRzFTTFfQF7ruMzaWPunDe6Tu9YxDJ2pD9W3I+BsUwz4JFylLSb1NbV5Y1qFqa1jK6YE5BUUAa9EVhASSCrla6W/fKuGhTPt6o7J7paQwTF6sqlqLCbo9YrFe2LwGUS/eGKK8kO3I8ZVczxOMyA1VvnaLJ+c9MHHSw= } ; } } 8, 18 -- Message-ID: } {20060814113015.81604.qmail-at-web37406.mail.mud.yahoo.com} } } 8, 18 -- Received: from [80.122.101.102] by } web37406.mail.mud.yahoo.com via HTTP; Mon, 14 Aug } 2006 04:30:15 PDT } } 8, 18 -- Date: Mon, 14 Aug 2006 04:30:15 -0700 } (PDT) } } 8, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } } 8, 18 -- Subject: need infos about e diffraction } in TEM } } 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============================== } } }
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==============================Original Headers============================== 9, 20 -- From nizets2-at-yahoo.com Thu Aug 17 01:48:40 2006 9, 20 -- Received: from web37408.mail.mud.yahoo.com (web37408.mail.mud.yahoo.com [209.191.91.140]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7H6mdnF020219 9, 20 -- for {microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 01:48:39 -0500 9, 20 -- Received: (qmail 65219 invoked by uid 60001); 17 Aug 2006 06:48:39 -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=46+tg4vtVqYk4X30p1FZiDj9JUtYOUZ70i9iC+IOR1ImeYHrIORqoXUOjWeXobDcVAU02OuMvDMZ4v7ppX2u88xgiQDYbIyQvQgXztDbAN583tchZWrl6Hg9mt/w7+WBRWOLe0BnTVxaZ/lKCbshR91+u4UvaKgZf7+dhg1OGIU= ; 9, 20 -- Message-ID: {20060817064839.65217.qmail-at-web37408.mail.mud.yahoo.com} 9, 20 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via HTTP; Wed, 16 Aug 2006 23:48:39 PDT 9, 20 -- Date: Wed, 16 Aug 2006 23:48:39 -0700 (PDT) 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 9, 20 -- Subject: Re: [Microscopy] need infos about e diffraction in TEM 9, 20 -- To: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} 9, 20 -- Cc: microscopy-at-microscopy.com 9, 20 -- In-Reply-To: {200608141344.k7EDiSc09038-at-www.matscieng.sunysb.edu} 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 get asked this fairly regularly by schools and parents (so we have stock replies). Often professional microscopists aren't the best source of information for cheap microscope's as all our optical systems are always over £10,000, and most lab mainstays like our three confocal microscopes come in at nearer £200,000 each (and users still complain about image quality). I assume you are asking about home use with your own kids.
Cheap microscopes under £100 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 and you may make an expensive mistake. Look for old branded 'laboratory' microscopes e.g. Bausch & Lomb, Prior, Leitz, Reichert, Baker, but probably not Tasco toys. Famous existing brands like Zeiss and Olympus attract a high premium. The sellers are often not microscopists though, and many are sold as collector's items and not for 'scientific' use. Also try any local microscope enthusiast clubs - they aren't as common as the many excellent astronomy [telescope] clubs but they are about and have knowledgeable enthusiasts with an eye for low cost quality systems.
There are suppliers geared up to providing cheaper microscopes for schools, so you can ask around at school's science departments, but expect to pay nearer £500 each for a quality setup (although with those like bottom end Meade [www.meade.com] at around £100 you can see something at low mag (~20x objective i.e. around 180x mag) with a quality stained section.
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 £1 and a good low mag Osram one that includes an illuminating halogen bulb at £8. In general I would say a good stereo dissecting microscope is a good choice for kids as it's great for viewing living things and enlarges what you can see already - look for 40x rather than 4x though.
Generally prepared slides can rapidly get very boring for under 14s, but living or unusual things (even hamster fur) always attract an audience. Also try your flatbed film scanner (not LiDe and from around £60 upwards) that will be good for looking at soft static things: leaves, fruit, nuts, household objects (scan at max resolution and try reflected and 'film' mode). In the UK there are sites like http://www.wedgwood-group.com/microscopes_digital.htm that cater for schools and colleges, providing standard compound and stereo microscopes as well as cheap PC video based microscope solutions where the whole class 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 via ebay, but they tend to be expensive and are easily broken by any age-group. Mounted slides keep well though, so 'vintage' ones even from 50 years ago can still look OK - most schools will have some specimen slides knocking about.
At home and our Primary school (under 11) we use the Digital Blue QX-5 (£70) - it's fun but pretty useless for serious microscope work as it's so low res (but at 640 x 480 better than the old Intel QX-3 it replaced). It puts the image on a PC screen. I have one at home for my kids (boy 10 and girl 12) but it only gets occasional use now the novelty has worn off. See http://micro.magnet.fsu.edu/optics/intelplay/index.html (not updated for the QX-5 but all applies - the site even discusses ways of contrast enhancement etc..). Once on the PC the 640x480 images can be manipulated and pasted etc, and the QX-5 does time-lapse for things like crystal growth. living plants growing and small animals. The similar but far better built Olympus MIC-D was great but being over £500 it was just too expensive for most schools and is now discontinued - there are other similar budget systems about though.
The macro on a good digital camera (like the image stabilised 5MP Canon S2IS going cheap at £200 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. Plus you can the camera for normal photography when you bored with microscopy.
By the way, if you get a microscope, 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: schooley-at-mcn.org [mailto:schooley-at-mcn.org] Sent: 16 August 2006 05:55 To: keith.morris-at-ucl.ac.uk
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I'll be happy to provide specific advice, but I need to know more about your needs & experience. I'm reluctant to recommend a used scope unless you have enough experience to recognize problems, and to fix them. Does "plant and animal cells" mean purchased, prepared slides? And what do you mean by "dissected samples"? What is your budget?
Although the microscope-buying advice on the MICRO website (URL below) is intended for a younger age group, you may find it a useful place to begin. There are several books in the MICRO bibliography that can help; again, I need to know more to make suggestions.
Caroline
-- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 5, 19 -- From schooley-at-mcn.org Tue Aug 15 23:50:47 2006 5, 19 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7G4olg8016555 5, 19 -- for {microscopy-at-microscopy.com} ; Tue, 15 Aug 2006 23:50:47 -0500 5, 19 -- Received: from [66.52.139.254] (helo=[10.0.1.2]) 5, 19 -- by dns4.mcn.org with esmtpa (Exim 4.60) 5, 19 -- (envelope-from {schooley-at-mcn.org} ) 5, 19 -- id J42QSJ-000HO6-4P; Tue, 15 Aug 2006 21:50:44 -0700 5, 19 -- Mime-Version: 1.0 5, 19 -- Message-Id: {a06200700c1084e7f6931-at-[10.0.1.2]} 5, 19 -- In-Reply-To: {200608160122.k7G1MtaK013462-at-ns.microscopy.com} 5, 19 -- References: {200608160122.k7G1MtaK013462-at-ns.microscopy.com} 5, 19 -- Date: Tue, 15 Aug 2006 21:44:17 -0700 5, 19 -- To: jchampagne-at-caseforensicscorp.com 5, 19 -- From: Caroline Schooley {schooley-at-mcn.org} 5, 19 -- Subject: Re: [Microscopy] [Filtered] AskAMicroscopist: Used Microscopes 5, 19 -- with my high school 5, 19 -- Cc: microscopy-at-microscopy.com 5, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 33, 27 -- From keith.morris-at-ucl.ac.uk Thu Aug 17 04:23:33 2006 33, 27 -- Received: from vscani-a.ucl.ac.uk (vscani-a.ucl.ac.uk [144.82.108.29]) 33, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7H9NWRx000615 33, 27 -- for {Microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 04:23:32 -0500 33, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 33, 27 -- by vscani-a.ucl.ac.uk with esmtp (Exim 4.60) 33, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 33, 27 -- id 1GDe64-0001dL-MG; Thu, 17 Aug 2006 10:23:28 +0100 33, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 33, 27 -- To: {jchampagne-at-caseforensicscorp.com} 33, 27 -- Cc: {Microscopy-at-microscopy.com} 33, 27 -- Subject: RE: [Microscopy] Re: [Filtered] AskAMicroscopist: Used Microscopes 33, 27 -- Date: Thu, 17 Aug 2006 10:23:27 +0100 33, 27 -- Message-ID: {001501c6c1de$cc1cfc70$7b865290-at-keithhigrade} 33, 27 -- MIME-Version: 1.0 33, 27 -- Content-Type: text/plain; 33, 27 -- charset="iso-8859-1" 33, 27 -- X-Mailer: Microsoft Office Outlook 11 33, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 33, 27 -- Thread-Index: AcbA8CYRO86f57EFRpe8Ilig1fK6ogA6gu0g 33, 27 -- In-Reply-To: {200608160455.k7G4t0xa023549-at-ns.microscopy.com} 33, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 33, 27 -- X-UCL-MailScanner: Found to be clean 33, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 33, 27 -- X-Spam-Status: No 33, 27 -- Content-Transfer-Encoding: 8bit 33, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7H9NWRx000615 ==============================End of - Headers==============================
A cautionary note about buying lab equipment on e-bay,etc:
A few years ago, we had a rash of small and not-so-small instrumentation going missing (pipettors, and microscope components). An astute pot-doc who was playing "let's see if e-bay any any _____" came across listings of items for sale, all by one seller, that very closely matched the missing material. He reported it to the med school's security department who investigated. It turned out that another post-doc had been spiriting items out of his lab and the labs of his PI's collaborators and supplementing his income by selling them on e-bay. e-bay was EXTREMELY cooperative about helping with the investigation and, needless to say that fellow's career was brought to a crashing halt. e-bay has grown so large that although it makes efforts to control what's on its site, clearly, it cannot do so 100%. I don't know the final outcome with regard to the stolen materials.
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============================== 3, 23 -- From lcgould-at-med.cornell.edu Thu Aug 17 08:15:40 2006 3, 23 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7HDFdPY030757 3, 23 -- for {microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 08:15:40 -0500 3, 23 -- Received: from mpx2.med.cornell.edu (pc113142-10.med.cornell.edu [140.251.11.119]) 3, 23 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7HDFZx9027395 3, 23 -- for {microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 09:15:36 -0400 (EDT) 3, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 3, 23 -- by mpx2.med.cornell.edu 3, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 3, 23 -- with ESMTPA id {0J45003A78TZX220-at-mpx2.med.cornell.edu} for 3, 23 -- microscopy-at-microscopy.com; Thu, 17 Aug 2006 09:15:35 -0400 (EDT) 3, 23 -- Date: Thu, 17 Aug 2006 09:15:40 -0400 3, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 3, 23 -- Subject: Re: [Microscopy] [Filtered] AskAMicroscopist: Used Microscopes 3, 23 -- In-reply-to: {200608170924.k7H9OKKr001891-at-ns.microscopy.com} 3, 23 -- Sender: lcgould-at-med.cornell.edu 3, 23 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 3, 23 -- Message-id: {p06230900c10a1a54da70-at-[140.251.48.23]} 3, 23 -- MIME-version: 1.0 3, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 3, 23 -- References: {200608170924.k7H9OKKr001891-at-ns.microscopy.com} 3, 23 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.8.17.55442 ==============================End of - Headers==============================
Has anyone used this particular microscope EM-109 (Zeiss) and have had any problems with it? I can't seem to get the filament to work, and the condenser moves when I go to spread the beam out. Anyone have any suggestions as to how to fix it?
Yarn Dept of Agriculture
==============================Original Headers============================== 3, 19 -- From addlvirology-at-mail.agri.state.oh.us Thu Aug 17 08:36:10 2006 3, 19 -- Received: from ODANTMAIL.agri.state.oh.us (mail.agri.state.oh.us [198.234.149.236]) 3, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7HDa97f011571 3, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 08:36:09 -0500 3, 19 -- Received: from 198.234.153.194 by ODANTMAIL.agri.state.oh.us (InterScan E-Mail VirusWall NT); Thu, 17 Aug 2006 09:30:54 -0400 3, 19 -- Received: from EAG13221 [172.16.6.11] by mail.agri.state.oh.us with ESMTP 3, 19 -- (SMTPD-8.21) id A1460200; Thu, 17 Aug 2006 09:38:14 -0400 3, 19 -- From: "addlvirology" {addlvirology-at-mail.agri.state.oh.us} 3, 19 -- To: {Microscopy-at-microscopy.com} 3, 19 -- Subject: EM-109 3, 19 -- Date: Thu, 17 Aug 2006 09:35:59 -0400 3, 19 -- MIME-Version: 1.0 3, 19 -- Content-Type: text/plain; 3, 19 -- charset="us-ascii" 3, 19 -- Content-Transfer-Encoding: 7bit 3, 19 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 3, 19 -- Thread-Index: AcbCAhL/QlttoiuUQ2CApHB7tMXKHA== 3, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1165 3, 19 -- Message-Id: {200608170938156.SM02148-at-EAG13221} ==============================End of - Headers==============================
Regards purchase on eBay of 'hot' items, always ask the seller for serial number information. This can provide information about the age and legitimacy of the item. If the seller will not provide information, it's probably best to not purchase.
As a further cautionary note - you have no guarantee that the equipment you purchase will work properly. Make sure that the seller is reputable and that you can service or repair whatever you buy. We recently purchased a microcentrifuge using eBay to replace equipment which went to another lab - totally legitimately, the other person had paid for it and the group was growing. The microcentrifuge works well. As a result, I am now thinking about purchasing variable volume pipetters. Again, to replace equipment which was moved to the other lab. In the first case, I followed the caveats of getting a history before bidding, knowing what the item cost new, knowing what it may cost to repair or calibrate the equipment, and establishing a maximum amount I'm was willing to bid. I will do the same if we decide to actually purchase pipetters.
Finally, remember that in many ways eBay is a game where people may get caught up in the thrill of the chase, and most of that chase occurs in the last 10 minutes. Stick to that maximum you set or you may get caught up in the game and pay too much.
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 Thu Aug 17 09:46:21 2006 10, 20 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7HEkJ9Q025211 10, 20 -- for {microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 09:46:20 -0500 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 k7HEkHiZ009533; 10, 20 -- Thu, 17 Aug 2006 09:46:18 -0500 (CDT) 10, 20 -- Message-ID: {44E48135.2030802-at-umanitoba.ca} 10, 20 -- Date: Thu, 17 Aug 2006 09:46:13 -0500 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 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 10, 20 -- Subject: Re: [Microscopy] Re: [Filtered] AskAMicroscopist: Used Microscopes 10, 20 -- References: {200608171318.k7HDI9pv002426-at-ns.microscopy.com} 10, 20 -- In-Reply-To: {200608171318.k7HDI9pv002426-at-ns.microscopy.com} 10, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Does anyone know if dispersants used in making cement (yup...new sample for us biologists!) can act as cryo-protectants when freezing the sample using HPS? The dispersants are proprietary comb-type polycarboxylates with straight backbones and parallel short chains extending outward.
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============================== 4, 21 -- From dsherman-at-purdue.edu Thu Aug 17 10:26:48 2006 4, 21 -- Received: from 1061exfe03.adpc.purdue.edu (1061exfe03.adpc.purdue.edu [128.210.63.225]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7HFQma5007928 4, 21 -- for {microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 10:26:48 -0500 4, 21 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe03.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 4, 21 -- Thu, 17 Aug 2006 11:26:47 -0400 4, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 4, 21 -- Thu, 17 Aug 2006 15:26:47 +0000 4, 21 -- User-Agent: Microsoft-Entourage/11.2.5.060620 4, 21 -- Date: Thu, 17 Aug 2006 11:26:46 -0400 4, 21 -- Subject: Cryo-protectant for cement 4, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 4, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 4, 21 -- Message-ID: {C10A02F6.137B7%dsherman-at-purdue.edu} 4, 21 -- Thread-Topic: Cryo-protectant for cement 4, 21 -- Thread-Index: AcbCEYzay2zl1C4EEduGmQARJN08Mg== 4, 21 -- Mime-version: 1.0 4, 21 -- Content-type: text/plain; 4, 21 -- charset="US-ASCII" 4, 21 -- Content-transfer-encoding: 7bit 4, 21 -- X-OriginalArrivalTime: 17 Aug 2006 15:26:47.0993 (UTC) FILETIME=[8E0A9A90:01C6C211] ==============================End of - Headers==============================
The University of Oregon is hosting a workshop for Energy Dispersive Analysis (EDS) which will focus of quantitative phase mapping and low voltage (high resolution) analysis of materials on the scanning electron microscope and electron microprobe analyzer. Topics will include identification of minor and trace phases and contaminates, phase distribution and modal abundance and quantitation of small particles and other embedded phases. This workshop will be a hands on exploration of the state of the art on several instruments offering the latest instrumental breakthroughs for quantitative analysis and x-ray mapping capabilities with one of the world's leading experts: Dale Newbury from the National Institute of Standards and Technology.
1. Zeiss Ultra with Oxford Inca 2. FEI Quanta with Bruker XFlash 3. Cameca SX100 with Thermo System Six
Attendance is limited! Additional morning presentations are welcome- please contact John Donovan directly.
EDS Spectrum Imaging and Low Voltage (High Resolution) Analysis, University of Oregon, Eugene, OR
3 day workshop by Dale Newbury (National Institute of Standards & Technology), September, 12-14, 2006
Morning 1 talk: "Silicon Drift Detectors: Energy Dispersive X-ray Spectrometry and X-ray Spectrum Imaging at Output Count Rates Above 100 kHz, and What to Do with All This Data"
Afternoon Demos: * Spectral Imaging and Mapping with a conventional EDS (Oxford and Thermo) * Bruker Quad SDD and Thermo SDD doing high speed x-ray spectrum image mapping on an FEI Quanta SEM * LISPIX hands-on software lab, with pre-recorded x-ray spectrum image data sets.
Morning 2 talk: "Challenges to Successful EDS X-ray Microanalysis in the Low Beam Energy (E0 { 5 keV) Regime"
Afternoon demos: * Low Beam Energy SEM/EDS on selected specimens, e.g., SiC, BaTiO3, etc. * Demonstration of conventional WDS in the low beam energy regime with BaTiO3 scanning the TiL-O K and BaM spectral regions.
Morning 3 talk: "The Perils of Automatic EDS Analysis: Blunders in Automated Peak Identification of Major, Minor, and Trace Constituents and the Reality of Standardless Analysis"
Afternoon demo: * Testing Automatic Peak ID and Standardless Analysis with commercial software in the Laboratory * User suggestions for testing (bring your own samples)
Check this page for registration and further details: http://epmalab.uoregon.edu/eds_workshop.htm
Respond to John Donovan (donovan-at-uoregon.edu) to confirm your participation. Registration is required by September 1, 2006. More to come...
==============================Original Headers============================== 15, 21 -- From donovan-at-uoregon.edu Thu Aug 17 16:30:24 2006 15, 21 -- Received: from smtp.uoregon.edu (mserv3.uoregon.edu [128.223.142.101]) 15, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7HLUNwO026397 15, 21 -- for {Microscopy-at-Microscopy.Com} ; Thu, 17 Aug 2006 16:30:23 -0500 15, 21 -- Received: from Probev.uoregon.edu (probev.uoregon.edu [128.223.92.75]) 15, 21 -- (authenticated bits=0) 15, 21 -- by smtp.uoregon.edu (8.13.7/8.13.7) with ESMTP id k7HLUNtO031060 15, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT); 15, 21 -- Thu, 17 Aug 2006 14:30:23 -0700 15, 21 -- Message-Id: {7.0.1.0.0.20060817142610.03841d88-at-uoregon.edu} 15, 21 -- Message-Id: {7.0.1.0.0.20060817142352.03841268-at-uoregon.edu} 15, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 15, 21 -- Date: Thu, 17 Aug 2006 14:30:16 -0700 15, 21 -- To: Microscopy-at-Microscopy.Com 15, 21 -- From: John Donovan {donovan-at-uoregon.edu} 15, 21 -- Subject: EDS/Spectrum Imaging/Low Voltage Analysis Workshop, Eugene, OR 15, 21 -- Cc: dale.newbury-at-nist.gov 15, 21 -- Mime-Version: 1.0 15, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 15, 21 -- X-Virus-Scanned: ClamAV 0.88.4/1677/Thu Aug 17 06:56:09 2006 on mserv3 15, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
We are looking for opportunities to learn the techniques of electron diffractometry (by TEM). We are working with a Tecnai G20 and the main purpose is the analysis of natural mineral samples. We will consider any proposition from workshop to on-site training. A very important limitation is geography: we have no desire to leave our good old Europa (we are based in Austria).
Stephane
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==============================Original Headers============================== 5, 18 -- From nizets2-at-yahoo.com Fri Aug 18 01:57:03 2006 5, 18 -- Received: from web37414.mail.mud.yahoo.com (web37414.mail.mud.yahoo.com [209.191.91.146]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7I6v3oZ013219 5, 18 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 01:57:03 -0500 5, 18 -- Received: (qmail 98216 invoked by uid 60001); 18 Aug 2006 06:57:02 -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=iabXjwC63yxnnEBcw+wnNbXh+IQ6RnQrzLu5jCuhyfBKcC37mcdH9DhbO72LON/aB66wEAa8lG4nhjXsxw4ZnLG+a33tSFw2B+11LRWijaEOH/q7+Z72n1LPfThhlHaXUIbAUH/qpHLs9vLZ7fZWoUMOWrAASul0enA7ici0XhA= ; 5, 18 -- Message-ID: {20060818065702.98214.qmail-at-web37414.mail.mud.yahoo.com} 5, 18 -- Received: from [80.122.101.102] by web37414.mail.mud.yahoo.com via HTTP; Thu, 17 Aug 2006 23:57:02 PDT 5, 18 -- Date: Thu, 17 Aug 2006 23:57:02 -0700 (PDT) 5, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 5, 18 -- Subject: workshop/courses about electron diffractometry 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==============================
Not sure which Hirsch book jim had in mind but maybe one of :
TOPICS IN ELECTRON DIFFRACTION AND MICROSCOPY OF MATERIALS (1999) Electron Microscopy of Thin Crystals (1965)
(transmission isn't in the title..)
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 } ---------------------------------------------------------------------------- } } Thanks for the tip, but this book is not available at } Amazon. I did a search on the www but I cannot find a } distributor (it is probably too old). } } I already spent one day on the link given by Sousan } (very nice). } } Would there be another good reference book (more } recent perhaps) which does the same job as Hirsch's? } } Regards, } } Stephane } } --- Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} wrote: } } } Stephane } } } } Purchase a copy of Hirsch's book. } } Go to Amazon.com. } } Search for "transmission electron hirsch". } } } } regards, } } } } Jim } } } } } From mail-at-ns.microscopy.com Mon Aug 14 07:30:49 } } 2006 } } } Date: Mon, 14 Aug 2006 06:33:44 -0500 } } } To: jquinn-at-www.matscieng.sunysb.edu } } } From: nizets2-at-yahoo.com } } } Reply-to: nizets2-at-yahoo.com } } } X-Resent-From: "Microscopy Listserver" } } {microscopy-at-microscopy.com} } } } Subject: [Microscopy] need infos about e } } diffraction in TEM } } } Errors-To: } } MicroscopyListSpamFilter-at-microscopy.com } } } X-lewp: MicroscopyListSpam NAGS } } } } } } } } } } } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The } } Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } ---------------------------------------------------------------------------- } } } } } } Dear listers, } } } } } } Although I understand the principle behind } } electron } } } diffraction in TEM I have no idea about the kind } } of } } } informations this technique gives and how to } } interpret } } } the diffraction pattern. } } } If you could give me a www address which explains } } that } } } I would be much grateful. } } } } } } Now a very practical problem could be perhaps } } used as } } } an example: we have a mixture of aluminosilicate } } } mineral (I have cut 70 nm sections) with approx. } } 90% } } } of mordenite and 10% quarz. } } } 1) Can I use e diffraction to distinguish both } } types } } } of particles? (to verify the purity of the } } powder) } } } 2) What kind of information about the cristal } } } structure can e diffraction give me in this case? } } } 3) Can I detect a change -and which change- to } } the } } } cristal structure using this technique if the } } mineral } } } is heat and treated with strong acids? (which } } actually } } } modifies the structure) } } } } } } We have also a tilt stage for tomography. Can } } this } } } bring further informations? } } } } } } Regards, } } } } } } Stephane } } } } } } } } } } } __________________________________________________ } } } Do You Yahoo!? } } } Tired of spam? Yahoo! Mail has the best spam } } protection around } } } http://mail.yahoo.com } } } } } } ==============================Original } } Headers============================== } } } 8, 18 -- From nizets2-at-yahoo.com Mon Aug 14 } } 06:30:17 2006 } } } 8, 18 -- Received: from } } web37406.mail.mud.yahoo.com } } (web37406.mail.mud.yahoo.com [209.191.91.138]) } } } 8, 18 -- by ns.microscopy.com } } (8.12.11.20060308/8.12.8) with SMTP id } } k7EBUGao010591 } } } 8, 18 -- for {microscopy-at-microscopy.com} ; Mon, } } 14 Aug 2006 06:30:17 -0500 } } } 8, 18 -- Received: (qmail 81606 invoked by uid } } 60001); 14 Aug 2006 11:30:15 -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=rlmt7902ZNRzFTTFfQF7ruMzaWPunDe6Tu9YxDJ2pD9W3I+BsUwz4JFylLSb1NbV5Y1qFqa1jK6YE5BUUAa9EVhASSCrla6W/fKuGhTPt6o7J7paQwTF6sqlqLCbo9YrFe2LwGUS/eGKK8kO3I8ZVczxOMyA1VvnaLJ+c9MHHSw= } } ; } } } 8, 18 -- Message-ID: } } } {20060814113015.81604.qmail-at-web37406.mail.mud.yahoo.com} } } } 8, 18 -- Received: from [80.122.101.102] by } } web37406.mail.mud.yahoo.com via HTTP; Mon, 14 Aug } } 2006 04:30:15 PDT } } } 8, 18 -- Date: Mon, 14 Aug 2006 04:30:15 -0700 } } (PDT) } } } 8, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } } } 8, 18 -- Subject: need infos about e diffraction } } in TEM } } } 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============================== } } } } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 9, 20 -- From nizets2-at-yahoo.com Thu Aug 17 01:48:40 2006 } 9, 20 -- Received: from web37408.mail.mud.yahoo.com (web37408.mail.mud.yahoo.com [209.191.91.140]) } 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7H6mdnF020219 } 9, 20 -- for {microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 01:48:39 -0500 } 9, 20 -- Received: (qmail 65219 invoked by uid 60001); 17 Aug 2006 06:48:39 -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=46+tg4vtVqYk4X30p1FZiDj9JUtYOUZ70i9iC+IOR1ImeYHrIORqoXUOjWeXobDcVAU02OuMvDMZ4v7ppX2u88xgiQDYbIyQvQgXztDbAN583tchZWrl6Hg9mt/w7+WBRWOLe0BnTVxaZ/lKCbshR91+u4UvaKgZf7+dhg1OGIU= ; } 9, 20 -- Message-ID: {20060817064839.65217.qmail-at-web37408.mail.mud.yahoo.com} } 9, 20 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via HTTP; Wed, 16 Aug 2006 23:48:39 PDT } 9, 20 -- Date: Wed, 16 Aug 2006 23:48:39 -0700 (PDT) } 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 9, 20 -- Subject: Re: [Microscopy] need infos about e diffraction in TEM } 9, 20 -- To: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} } 9, 20 -- Cc: microscopy-at-microscopy.com } 9, 20 -- In-Reply-To: {200608141344.k7EDiSc09038-at-www.matscieng.sunysb.edu} } 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============================== }
--
Andy Buckley
AB78-at-ESC.CAM.AC.UK DEPARTMENT OF EARTH SCIENCES UNIVERSITY OF CAMBRIDGE Tel +44 1223 333469 DOWNING STREET +44 1223 333400 CAMBRIDGE FAX +44 1223 333450 CB2 3EQ
Manager of XRD discussion list: http://www.jiscmail.ac.uk/lists/xrd.html
==============================Original Headers============================== 10, 22 -- From ab78-at-esc.cam.ac.uk Fri Aug 18 04:04:14 2006 10, 22 -- Received: from rock.esc.cam.ac.uk (rock.esc.cam.ac.uk [131.111.41.250]) 10, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7I94EdF025536 10, 22 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 04:04:14 -0500 10, 22 -- Received: from andyb.esc.cam.ac.uk ([192.168.17.155]) 10, 22 -- by rock.esc.cam.ac.uk with esmtps (TLSv1:AES256-SHA:256) 10, 22 -- (Exim 4.60) 10, 22 -- (envelope-from {ab78-at-esc.cam.ac.uk} ) 10, 22 -- id 1GE0Gv-0007Xu-UX; Fri, 18 Aug 2006 10:04:13 +0100 10, 22 -- Message-ID: {44E5824A.8000803-at-esc.cam.ac.uk} 10, 22 -- Date: Fri, 18 Aug 2006 10:03:06 +0100 10, 22 -- From: Andy Buckley {ab78-at-esc.cam.ac.uk} 10, 22 -- Reply-To: ab78-at-esc.cam.ac.uk 10, 22 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 10, 22 -- MIME-Version: 1.0 10, 22 -- To: nizets2-at-yahoo.com, microscopy-at-microscopy.com, 10, 22 -- jquinn-at-www.matscieng.sunysb.edu 10, 22 -- Subject: Re: [Microscopy] Re: need infos about e diffraction in TEM 10, 22 -- References: {200608170654.k7H6sQM9026028-at-ns.microscopy.com} 10, 22 -- In-Reply-To: {200608170654.k7H6sQM9026028-at-ns.microscopy.com} 10, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 22 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I was able to obtain a reprint of ELECTRON MICROSCOPY OF THIN CRYSTALS by Hirsch, et al. from Krieger Publishing Company, Malabar, Florida
Cheers
Roger A. Ristau, PhD Electron Microscopy Specialist Institute of Materials Science 97 North Eagleville Road University of Connecticut Storrs, CT 06269 vox: 860-486-5379 fax: 860-486-4745
} From: nizets2-at-yahoo.com } Reply-To: nizets2-at-yahoo.com } Date: Thu, 17 Aug 2006 01:57:38 -0500 } To: raristau-at-ims.uconn.edu } Subject: [Microscopy] Re: need infos about e diffraction in 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 } ---------------------------------------------------------------------------- } } Thanks for the tip, but this book is not available at } Amazon. I did a search on the www but I cannot find a } distributor (it is probably too old). } } I already spent one day on the link given by Sousan } (very nice). } } Would there be another good reference book (more } recent perhaps) which does the same job as Hirsch's? } } Regards, } } Stephane } } --- Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} wrote: } } } Stephane } } } } Purchase a copy of Hirsch's book. } } Go to Amazon.com. } } Search for "transmission electron hirsch". } } } } regards, } } } } Jim } } } } } From mail-at-ns.microscopy.com Mon Aug 14 07:30:49 } } 2006 } } } Date: Mon, 14 Aug 2006 06:33:44 -0500 } } } To: jquinn-at-www.matscieng.sunysb.edu } } } From: nizets2-at-yahoo.com } } } Reply-to: nizets2-at-yahoo.com } } } X-Resent-From: "Microscopy Listserver" } } {microscopy-at-microscopy.com} } } } Subject: [Microscopy] need infos about e } } diffraction in TEM } } } Errors-To: } } MicroscopyListSpamFilter-at-microscopy.com } } } X-lewp: MicroscopyListSpam NAGS } } } } } } } } } } } } } } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The } } Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } } On-Line Help } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } } } } } ---------------------------------------------------------------------------- } } } } } } Dear listers, } } } } } } Although I understand the principle behind } } electron } } } diffraction in TEM I have no idea about the kind } } of } } } informations this technique gives and how to } } interpret } } } the diffraction pattern. } } } If you could give me a www address which explains } } that } } } I would be much grateful. } } } } } } Now a very practical problem could be perhaps } } used as } } } an example: we have a mixture of aluminosilicate } } } mineral (I have cut 70 nm sections) with approx. } } 90% } } } of mordenite and 10% quarz. } } } 1) Can I use e diffraction to distinguish both } } types } } } of particles? (to verify the purity of the } } powder) } } } 2) What kind of information about the cristal } } } structure can e diffraction give me in this case? } } } 3) Can I detect a change -and which change- to } } the } } } cristal structure using this technique if the } } mineral } } } is heat and treated with strong acids? (which } } actually } } } modifies the structure) } } } } } } We have also a tilt stage for tomography. Can } } this } } } bring further informations? } } } } } } Regards, } } } } } } Stephane } } } } } } } } } } } __________________________________________________ } } } Do You Yahoo!? } } } Tired of spam? Yahoo! Mail has the best spam } } protection around } } } http://mail.yahoo.com } } } } } } ==============================Original } } Headers============================== } } } 8, 18 -- From nizets2-at-yahoo.com Mon Aug 14 } } 06:30:17 2006 } } } 8, 18 -- Received: from } } web37406.mail.mud.yahoo.com } } (web37406.mail.mud.yahoo.com [209.191.91.138]) } } } 8, 18 -- by ns.microscopy.com } } (8.12.11.20060308/8.12.8) with SMTP id } } k7EBUGao010591 } } } 8, 18 -- for {microscopy-at-microscopy.com} ; Mon, } } 14 Aug 2006 06:30:17 -0500 } } } 8, 18 -- Received: (qmail 81606 invoked by uid } } 60001); 14 Aug 2006 11:30:15 -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-T } ransfer-Encoding; } } } 8, 18 -- } } } b=rlmt7902ZNRzFTTFfQF7ruMzaWPunDe6Tu9YxDJ2pD9W3I+BsUwz4JFylLSb1NbV5Y1qFqa1jK6Y } E5BUUAa9EVhASSCrla6W/fKuGhTPt6o7J7paQwTF6sqlqLCbo9YrFe2LwGUS/eGKK8kO3I8ZVczxOM } yA1VvnaLJ+c9MHHSw= } } ; } } } 8, 18 -- Message-ID: } } } {20060814113015.81604.qmail-at-web37406.mail.mud.yahoo.com} } } } 8, 18 -- Received: from [80.122.101.102] by } } web37406.mail.mud.yahoo.com via HTTP; Mon, 14 Aug } } 2006 04:30:15 PDT } } } 8, 18 -- Date: Mon, 14 Aug 2006 04:30:15 -0700 } } (PDT) } } } 8, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } } } 8, 18 -- Subject: need infos about e diffraction } } in TEM } } } 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============================== } } } } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 9, 20 -- From nizets2-at-yahoo.com Thu Aug 17 01:48:40 2006 } 9, 20 -- Received: from web37408.mail.mud.yahoo.com } (web37408.mail.mud.yahoo.com [209.191.91.140]) } 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } k7H6mdnF020219 } 9, 20 -- for {microscopy-at-microscopy.com} ; Thu, 17 Aug 2006 01:48:39 -0500 } 9, 20 -- Received: (qmail 65219 invoked by uid 60001); 17 Aug 2006 06:48:39 } -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=46+tg4vtVqYk4X30p1FZiDj9JUtYOUZ70i9iC+IOR1ImeYHrIORqoXUOjWeXobDcVAU02OuMvDMZ } 4v7ppX2u88xgiQDYbIyQvQgXztDbAN583tchZWrl6Hg9mt/w7+WBRWOLe0BnTVxaZ/lKCbshR91+u4 } UvaKgZf7+dhg1OGIU= ; } 9, 20 -- Message-ID: {20060817064839.65217.qmail-at-web37408.mail.mud.yahoo.com} } 9, 20 -- Received: from [80.122.101.102] by web37408.mail.mud.yahoo.com via } HTTP; Wed, 16 Aug 2006 23:48:39 PDT } 9, 20 -- Date: Wed, 16 Aug 2006 23:48:39 -0700 (PDT) } 9, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 9, 20 -- Subject: Re: [Microscopy] need infos about e diffraction in TEM } 9, 20 -- To: Jim Quinn {jquinn-at-www.matscieng.sunysb.edu} } 9, 20 -- Cc: microscopy-at-microscopy.com } 9, 20 -- In-Reply-To: {200608141344.k7EDiSc09038-at-www.matscieng.sunysb.edu} } 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==============================
==============================Original Headers============================== 7, 20 -- From raristau-at-ims.uconn.edu Fri Aug 18 09:02:32 2006 7, 20 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7IE2T5i008464 7, 20 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 09:02:31 -0500 7, 20 -- Received: from [137.99.20.210] (d20h210.public.uconn.edu [137.99.20.210]) 7, 20 -- by mail2.uits.uconn.edu (8.13.6/8.11.6) with ESMTP id k7IE2NSh029812; 7, 20 -- Fri, 18 Aug 2006 10:02:23 -0400 7, 20 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 7, 20 -- Date: Fri, 18 Aug 2006 10:01:55 -0400 7, 20 -- Subject: Re: [Microscopy] Re: need infos about e diffraction in TEM 7, 20 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 7, 20 -- To: {nizets2-at-yahoo.com} , {microscopy-at-microscopy.com} 7, 20 -- Message-ID: {C10B4093.13EF%raristau-at-ims.uconn.edu} 7, 20 -- In-Reply-To: {200608170657.k7H6vcJb028789-at-ns.microscopy.com} 7, 20 -- Mime-version: 1.0 7, 20 -- Content-type: text/plain; charset="US-ASCII" 7, 20 -- Content-transfer-encoding: 7bit 7, 20 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 7, 20 -- X-UConn-MailScanner: Found to be clean 7, 20 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
We are looking for solid state secondary electron detector that is similar to Si PIN type, but more sensitive (higher gain) than Si (not scintillator type (E-T type).
Joe Wang
Project Manager Hermes Microvision, Inc. 1595 McVandless Drive Milpitas, CA 95035 e-Mail: joe.wang-at-hermes-microvision.com
Office: (408)273-5855
==============================Original Headers============================== 5, 24 -- From joe.wang-at-hermes-microvision.com Fri Aug 18 12:10:48 2006 5, 24 -- Received: from mail.hermes-microvision.com (mail.hermes-microvision.com [65.244.115.229]) 5, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7IHAk8M021826 5, 24 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 12:10:47 -0500 5, 24 -- Received: from hmilap108 ([10.200.98.18]) 5, 24 -- (authenticated bits=0) 5, 24 -- by mail.hermes-microvision.com (8.12.11/8.12.11) with ESMTP id k7IHAeaa009721 5, 24 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 10:10:43 -0700 5, 24 -- Message-Id: {200608181710.k7IHAeaa009721-at-mail.hermes-microvision.com} 5, 24 -- From: "Joe Wang" {joe.wang-at-hermes-microvision.com} 5, 24 -- To: {microscopy-at-microscopy.com} 5, 24 -- Subject: Secondary electron detector 5, 24 -- Date: Fri, 18 Aug 2006 10:07:56 -0700 5, 24 -- Organization: HMI 5, 24 -- MIME-Version: 1.0 5, 24 -- Content-Type: text/plain; 5, 24 -- charset="us-ascii" 5, 24 -- Content-Transfer-Encoding: 7bit 5, 24 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 5, 24 -- Thread-Index: AcbC6Nmt/yFpA4h5QTyYYNWtr4lquQ== 5, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 5, 24 -- X-Hermes-Microvision.-MailScanner-Information: Scanned at Hermes Microvision for your protection 5, 24 -- X-Hermes-Microvision.-MailScanner: Found to be clean 5, 24 -- X-MailScanner-From: joe.wang-at-hermes-microvision.com ==============================End of - Headers==============================
For a few days I have been trying to post a message that we are giving away our JEOL 100CX, but I guess it was always filtered as SPAM. So here it is again:
We are giving away our JEOL 100CX STEM. It is in running condition (or I should say was until about 9 months ago, we haven't used it since). We will keep the chiller. The new owner will be responsible for packing and shipping. The STEM is located in Lakewood, Colorado. No warranties or guarantees.
Let me know if you are interested. I'll make a decision next week.
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 Headers============================== 10, 23 -- From Mike.Bode-at-olympus-sis.com Fri Aug 18 17:17:57 2006 10, 23 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7IMHu4O005936 10, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 17:17:56 -0500 10, 23 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 10, 23 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id k7IMI5fV012729 10, 23 -- for {Microscopy-at-microscopy.com} ; Sat, 19 Aug 2006 00:18:07 +0200 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: FW: Anybody interested in a JEOL 100CX 10, 23 -- Date: Sat, 19 Aug 2006 00:15:55 +0200 10, 23 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC9442B0F0-at-ms-s-gws.soft-imaging.net} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: Anybody interested in a JEOL 100CX 10, 23 -- Thread-Index: AcbDEmrptU3OlfipRl+LPzeOmwR//gAAWgPQ 10, 23 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 10, 23 -- To: {Microscopy-at-microscopy.com} 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7IMHu4O005936 ==============================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 zzhang-at-uwyo.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: zzhang-at-uwyo.edu Name: Zhaojie Zhang
Organization: University of Wyoming
Title-Subject: [Filtered] Question-Immuno-gold test
Question: Dear Listers:
I am having a problem with the secondary antibody - Immuno-gold conjugate (6 nm from Electron Microscopy Sciences). Here is what I did -
I run a test with a LR White embeded sample following the "standard" protocol for immuno-gold labeling. I could not find any gold particles - no labeling, no background. Trying to figure out what is wrong, I tested the secondary antibody alone - put a small drop of the original antibody (no dilution) directly onto a TEM grid, air dry and viewed with TEM. I could not find any 6 nm gold particles. I did find a few particles ranging from 50 nm to 150 nm.
I then called the company, explained my problem. I was told that that is NOT the way I should test the secondary antibody, instead, I should do a dot-spot test (with silver enhancement?)
My Question is: Has anyone had this problem before? How do you check if the secondary antibody (immuno-gold) works or not?
Thank you,
Zhaojie Zhang Microscopy Core Facility University of Wyoming
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Email: KLRadke-at-ucdavis.edu Name: Kathryn Radke
Organization: Univ. California, Davis
Title-Subject: [Filtered] LM - coupling digital camera back to Nikon Diaphot
Question: I'd like to use my Nikon D70s camera back on my lab's 1985 Nikon Diaphot inverted cell culture microscope. The Diaphot front port has a fitting that couples to Nikon film backs (e.g. FE2, FTN) having Nikon F bayonet type lens mount. When the microscope is set up properly, the image viewed through the eyepieces focused on the film.
The D70S back won't fit directly onto the Diaphot. A local camera store tells me that I could use a T mount adapter that is designed to couple a digital back with a telescope. With the T mount adapter in place, the digital back can be used in Manual mode and the shutter speed can be set using the "command dial" on the camera back. I would also need to purchase a remote cord to operate the back without shaking the microscope.
I would be most grateful if anyone can give some feedback about necessary parts, operation, etc. Are there websites that have relevant information?
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Judy Murphy, PhD Microscopy Training, Imaging, and Lab Design Stockton, CA 95219 murphyjudy-at-comcast.net
connie.a.cummings-at-gsk.com wrote: Organization: GSK --| --|Title-Subject: [Filtered] EM tech certification --| --|Question: I was wondering where do you get information on certification on EM certification. --|
==============================Original Headers============================== 10, 17 -- From murphyjudy-at-comcast.net Fri Aug 18 18:30:31 2006 10, 17 -- Received: from alnrmhc11.comcast.net (alnrmhc14.comcast.net [206.18.177.54]) 10, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7INUVZY024544 10, 17 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 18:30:31 -0500 10, 17 -- Received: from [192.168.1.106] (c-67-181-78-13.hsd1.ca.comcast.net[67.181.78.13]) 10, 17 -- by comcast.net (alnrmhc14) with ESMTP 10, 17 -- id {20060818233030b1400gl0sve} ; Fri, 18 Aug 2006 23:30:31 +0000 10, 17 -- Message-ID: {44E64D94.1030204-at-comcast.net} 10, 17 -- Date: Fri, 18 Aug 2006 16:30:28 -0700 10, 17 -- From: Judy Murphy {murphyjudy-at-comcast.net} 10, 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 10, 17 -- X-Accept-Language: en-us, en 10, 17 -- MIME-Version: 1.0 10, 17 -- To: microscopy-at-microscopy.com 10, 17 -- Subject: Re: viaWWW: EM tech certification 10, 17 -- Content-Type: text/plain; charset=us-ascii; format=flowed 10, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
There are two tests you can do when you don't get a positive result im immuno gold labelling and when you want to check whether the conjugate performs up to standards:
1. An activity test, using a dot-spot system In this test a dilution series of corresponding IgG (Rabbit IgG in the present case) is spotted on a strip of nitrocellulose, and after blocking the strip is incubated with the gold conjugate. Silver enhancement is only required if you would test an ultra small particle conjugate in which the gold does not significantly contribute to otain colored dots. A 6nm conjugate has 'sufficient color' and does not need enhancement. We can supply you with a procedure if you like, they are described extensively in our Newsletter 4. 2. A TEM test in which the gold conjugate is adsorbed (not dried from the stock solution) onto a grid that is filmed and coated with poly-L- lysine which by its positive charge will bind negatively charged gold particles. Grids are washed on distilled water after adsorption. Again, we can supply you with a procedure if you like.
Drying a small drop of undiluted conjugated onto a filmed grid makes it not easy, if not impossible, to see particles or to evaluate what particle sizes you have. After all there is buffer components and protecting protein in the conjugate solution that all dry onto the grid. Also, upon drying particles tend to aggregate and form clumps (that are usually not easy to dissolve completely again, so they are pretty solid) and which will give erroneous readings.
I will be happy to help trying to stablish whether antigens may have been damaged preventing positive results or whether the primary antibody or secondary antibody have lost activity.
Cheers, hope this helps, get in touch if you like. New Zealand is still good fun!!
Jan Leunissen
Aurion - President Electron Microscopy Research Advisor Costerweg 5 Dept Anatomy and Structural Biology 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4795465 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz
==============================Original Headers============================== 10, 20 -- From leunissen-at-aurion.nl Fri Aug 18 18:43:14 2006 10, 20 -- Received: from fep06.xtra.co.nz (fep06.xtra.co.nz [210.54.141.240]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7INhDl6002485 10, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 18:43:14 -0500 10, 20 -- Received: from [192.168.1.50] (really [210.86.106.215]) by fep06.xtra.co.nz 10, 20 -- with ESMTP 10, 20 -- id {20060818234311.GTFE16670.fep06.xtra.co.nz-at-[192.168.1.50]} ; 10, 20 -- Sat, 19 Aug 2006 11:43:11 +1200 10, 20 -- In-Reply-To: {200608182305.k7IN5j1F017177-at-ns.microscopy.com} 10, 20 -- References: {200608182305.k7IN5j1F017177-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 10, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 10, 20 -- Message-Id: {86E9F1BD-5415-4582-AD4D-201906E99C1A-at-aurion.nl} 10, 20 -- Cc: Microscopy-at-microscopy.com 10, 20 -- Content-Transfer-Encoding: 7bit 10, 20 -- From: Jan Leunissen {leunissen-at-aurion.nl} 10, 20 -- Subject: Re: [Microscopy] viaWWW: Question-Immuno-gold test 10, 20 -- Date: Sat, 19 Aug 2006 11:43:10 +1200 10, 20 -- To: zzhang-at-uwyo.edu 10, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
I have a new topic that is not particularly relevant today but probably will be in the foreseeable future.
The newer SEMs are very advanced and make maximum use of PC interfacing. The PCs will age but should work OK or be replaced by faster models if necessary or due to supportability factors.
What about the SEM itself? These are now made with surface mount technology, flip chip BGAs and FPGAs.
Some makers are saying that they will support a specific product line for 5-10 more years, or less. After that, one needs to buy a new SEM. This is what they are saying....not all SEM makers. But enough to trigger the sensitivity of a potentially big bow wave of problems. The up-front cost of a tool really does not put it in the disposable category. Plus, in an industrial setting, how would depreciation factor in? But if the tool cannot be supported after say ten years, then it a disposable item if there are no other service resources. The dearth of schematics is a huge issue with me since I cannot talk to the service folks with knowledge and also impacts the future cut off of service. One is basically buying a $500K boat anchor that needs replacing every ten year or so. Yep, sounds like a boat problem.
Recall that the Amray and early JEOL tools were relatively easy to fix... electronics-wise. These newer systems are not at all easy to fix or troubleshoot.
I'm just blowing the whistle on what I think is a pending huge problem.
Alternate opinions?
gary g.
==============================Original Headers============================== 11, 17 -- From gary-at-gaugler.com Fri Aug 18 23:06:32 2006 11, 17 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7J46Vmm017237 11, 17 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 23:06:31 -0500 11, 17 -- Received: (qmail 17545 invoked from network); 18 Aug 2006 21:06:31 -0700 11, 17 -- Received: by simscan 1.1.0 ppid: 17542, pid: 17543, t: 0.1494s 11, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 17 -- by qsmtp2 with SMTP; 18 Aug 2006 21:06:30 -0700 11, 17 -- Message-Id: {7.0.1.0.2.20060818204828.0247a458-at-gaugler.com} 11, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 17 -- Date: Fri, 18 Aug 2006 21:06:33 -0700 11, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 11, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 17 -- Subject: New subject-service life of new SEMs 11, 17 -- Mime-Version: 1.0 11, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
All: Abebooks.com had several copies of both Hirsch books available, found by Advanced search on--- Author: hirsch Keywords: electron microscopy
-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
Gary, As a co-owner of a Jeol 6100 with an intermittent problem, and a soon-to-be owner of an 840F, I could not agree more. My 6100 does have schematics, but at the board level they are not even the correct boards. In defense of the manufacturers, they have high development costs and very low unit sales. Plus, the market size is very limited. If they were making a big profit, many other competitors would jump in.
I wonder why there are not specialty shops that take existing SEMs, strip them down to the column and other mechanical hardware, and couple up wholly new electronics. (Maybe there are. If so, I'd like to hear about them.) Isn't it just a more (ok, way more) complicated version of taking an old loudspeaker and hooking up a new amplifier?
With the technical service info secret, the customer is really leasing the tool, not buying it, since the customer is entirely dependent on the manufacturer to keep it running, and the manufacturer can define its useful life.
Is there a SEM manufacturer who sells complete service literature?
Bernard Cuzzillo Berkeley, CA USA
==============================Original Headers============================== 5, 14 -- From bernard-at-bearinc.com Sat Aug 19 00:59:21 2006 5, 14 -- Received: from bearinc.com (bearinc.com [64.62.217.44]) 5, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7J5xKvU006610 5, 14 -- for {Microscopy-at-microscopy.com} ; Sat, 19 Aug 2006 00:59:21 -0500 5, 14 -- Received: from 24.7.101.58 ([24.7.101.58]) by bearinc.com for {Microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 22:59:20 -0700 5, 14 -- Message-ID: {44E6A8B3.6030407-at-bearinc.com} 5, 14 -- Date: Fri, 18 Aug 2006 22:59:15 -0700 5, 14 -- From: Bernard Cuzzillo {bernard-at-bearinc.com} 5, 14 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 5, 14 -- MIME-Version: 1.0 5, 14 -- To: Microscopy-at-microscopy.com 5, 14 -- Subject: Service life of new SEMs -- Agreed! 5, 14 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 14 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
As one of the guys who can't afford the 500k for a new unit, I am looking forward to all of you dumping out of your 10 year old units when the time comes. My solution is simple, if the price is cheap enough you buy several for spares. Then you can remove-and-replace as required. I am currently keeping an old Kevex system running this way and have backup boards purchased for nothing on Ebay. I have also just purchased a complete JEOL 6100F for 800 bucks and am keeping an eye out for another. As a hobbyist this is my only choice so the less service the better for me.
On the other side of the coin, I would be blowing a fuse knowing my half mill investment would be worthless in 10 years. I agree with Gary that the schematics make all the difference.
Tom Kaye
==============================Original Headers============================== 5, 21 -- From tom-at-tomkaye.com Sat Aug 19 01:18:00 2006 5, 21 -- Received: from tomkaye.com (mxgateway.techpro.com [64.4.207.8]) 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7J6HxT6017057 5, 21 -- for {Microscopy-at-microscopy.com} ; Sat, 19 Aug 2006 01:18:00 -0500 5, 21 -- Received: from TKLAPTOP [65.39.118.10] by tomkaye.com with ESMTP 5, 21 -- (SMTPD32-8.14) id AD14220A00F4; Sat, 19 Aug 2006 01:17:56 -0500 5, 21 -- From: "Tom" {tom-at-tomkaye.com} 5, 21 -- To: {Microscopy-at-microscopy.com} 5, 21 -- Subject: RE: [Microscopy] Service life of new SEMs -- Agreed! 5, 21 -- Date: Sat, 19 Aug 2006 01:17:59 -0500 5, 21 -- Message-ID: {LHECKJAKKIOGKDBHHJMLMEDKCLAA.tom-at-tomkaye.com} 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 (Normal) 5, 21 -- X-MSMail-Priority: Normal 5, 21 -- X-Mailer: Microsoft Outlook IMO, Build 9.0.2416 (9.0.2911.0) 5, 21 -- In-reply-to: {200608190604.k7J646Pb014317-at-ns.microscopy.com} 5, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1165 5, 21 -- Importance: Normal ==============================End of - Headers==============================
The chair of the Microscopy Society of America Certification Board is E. Ann Ellis at eann.ellis-at-worldnet.att.com
Good Luck!
connie.a.cummings-at-gsk.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.com/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both connie.a.cummings-at-gsk.com as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: connie.a.cummings-at-gsk.com } Name: Connie Cummings } } Organization: GSK } } Title-Subject: [Filtered] EM tech certification } } Question: I was wondering where do you get information on certification on EM certification. } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 6, 12 -- From zaluzec-at-microscopy.com Fri Aug 18 18:06:15 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 k7IN6FvF018864 } 6, 12 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 18:06:15 -0500 } 6, 12 -- Mime-Version: 1.0 } 6, 12 -- X-Sender: (Unverified) } 6, 12 -- Message-Id: {p06110402c10bf85b4848-at-[206.69.208.22]} } 6, 12 -- Date: Fri, 18 Aug 2006 18:06:13 -0500 } 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: connie.a.cummings-at-gsk.com (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: EM tech certification } 6, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 4, 19 -- From randerson20-at-tampabay.rr.com Sat Aug 19 07:50:06 2006 4, 19 -- Received: from ms-smtp-05.tampabay.rr.com (ms-smtp-05.tampabay.rr.com [65.32.5.135]) 4, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7JCo6H7032338 4, 19 -- for {Microscopy-at-Microscopy.Com} ; Sat, 19 Aug 2006 07:50:06 -0500 4, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 4, 19 -- by ms-smtp-05.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k7JCo0WL023097; 4, 19 -- Sat, 19 Aug 2006 08:50:03 -0400 (EDT) 4, 19 -- Message-ID: {44E708F7.8070809-at-tampabay.rr.com} 4, 19 -- Date: Sat, 19 Aug 2006 08:49:59 -0400 4, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 4, 19 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 4, 19 -- MIME-Version: 1.0 4, 19 -- To: connie.a.cummings-at-gsk.com, Listserver {Microscopy-at-Microscopy.Com} 4, 19 -- Subject: Re: [Microscopy] viaWWW: EM tech certification 4, 19 -- References: {200608182306.k7IN6PG5019414-at-ns.microscopy.com} 4, 19 -- In-Reply-To: {200608182306.k7IN6PG5019414-at-ns.microscopy.com} 4, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 19 -- Content-Transfer-Encoding: 7bit 4, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (classyjay247-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Saturday, August 19, 2006 at 08:46:19 ---------------------------------------------------------------------------
Email: classyjay247-at-yahoo.com Name: Uchenna Victor
Organization: federal university of technology Owerri
Education: Undergraduate College
Location: Abuja Nigeria
Question: what are the transmission patterns of electron gun
Hirsch has also just listed at amazon.com along with several other diffraction books. Interesting tidbit: there are over 750 titles on electron microscopy on amazon!!! I was amazed. Below are a few with links on diffraction that one lister asked about. Cheers, Judy
Judy Murphy, PhD Microscopy Training, Imaging, and Lab Design Stockton, CA 95219 murphyjudy-at-comcast.net
Hirsch EM of Thin Crystals http://s1.amazon.com/exec/varzea/ts/exchange-glance/Y04Y5461265Y9057909/002-0880518-7016064
Modern Crystallography http://www.amazon.com/exec/obidos/ASIN/0387100520/002-0880518-7016064
Diffraction (Student Micrograph in Physics) http://www.amazon.com/exec/obidos/ASIN/0852745796/002-0880518-7016064
Andrews Interpretation of Electron Diffraction Patterns http://www.amazon.com/exec/obidos/ASIN/0306305348/002-0880518-7016064
McCall Interpretive Techniques for Microstructural Analysis http://www.amazon.com/exec/obidos/ASIN/0306310368/002-0880518-7016064
Rostoker Interpretation of Metallographic Structures http://www.amazon.com/exec/obidos/ASIN/0125982550/002-0880518-7016064
RMS Series Intro to Crystallography http://www.amazon.com/exec/obidos/ASIN/0198564333/002-0880518-7016064
Transmission Electron Microscopy of Minerals and Rocks http://www.amazon.com/exec/obidos/ASIN/0521350980/002-0880518-7016064
mike.reedy-at-cellbio.duke.edu wrote:
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==============================Original Headers============================== 15, 19 -- From murphyjudy-at-comcast.net Sat Aug 19 12:19:15 2006 15, 19 -- Received: from rwcrmhc15.comcast.net (rwcrmhc15.comcast.net [204.127.192.85]) 15, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7JHJFnF030017 15, 19 -- for {Microscopy-at-microscopy.com} ; Sat, 19 Aug 2006 12:19:15 -0500 15, 19 -- Received: from [192.168.1.106] (c-67-181-78-13.hsd1.ca.comcast.net[67.181.78.13]) 15, 19 -- by comcast.net (rwcrmhc15) with ESMTP 15, 19 -- id {20060819171913m1500jrcbte} ; Sat, 19 Aug 2006 17:19:14 +0000 15, 19 -- Message-ID: {44E74811.2040307-at-comcast.net} 15, 19 -- Date: Sat, 19 Aug 2006 10:19:13 -0700 15, 19 -- From: Judy Murphy {murphyjudy-at-comcast.net} 15, 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 15, 19 -- X-Accept-Language: en-us, en 15, 19 -- MIME-Version: 1.0 15, 19 -- To: mike.reedy-at-cellbio.duke.edu, Microscopy {Microscopy-at-microscopy.com} 15, 19 -- Subject: Re: [Microscopy] Re: need infos about e diffraction in TEM 15, 19 -- References: {200608190545.k7J5jNtW000349-at-ns.microscopy.com} 15, 19 -- In-Reply-To: {200608190545.k7J5jNtW000349-at-ns.microscopy.com} 15, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 15, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
You are right when you say that these are not disposable items. If manufacturers have the attitude that we in the academic world can simply conjure up $250-500,000 because they determine that the lifespan of their EM is 10 years, then the EM community collectively should send a strong message to the contrary. My attitude is this: if an EM manufacturer will not continue to support and keep my EM going until I am ready (and able) to replace it, then (when I do find the funds for a replacement) my next microscope will definitely not be one of theirs. There are companies that apparently stand up for their microscopes for the extended lifetimes they should be proud of. As an example, the lab I was trained in still has the microscope that I was trained on some 30 years ago (they added a digital camera, and I'm told it's one of the favored TEMs in the lab). --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 ---------------------------------------
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 } ---------------------------------------------------------------------------- } } Hi Listers: } } I have a new topic that is not particularly relevant } today but probably will be in the foreseeable future. } } The newer SEMs are very advanced and make maximum use } of PC interfacing. The PCs will age but should work OK } or be replaced by faster models if necessary or due to } supportability factors. } } What about the SEM itself? These are now made with } surface mount technology, flip chip BGAs and FPGAs. } } Some makers are saying that they will support a specific } product line for 5-10 more years, or less. After that, one needs } to buy a new SEM. This is what they are saying....not } all SEM makers. But enough to trigger the sensitivity of } a potentially big bow wave of problems. The up-front cost } of a tool really does not put it in the disposable category. } Plus, in an industrial setting, how would depreciation factor in? } But if the tool cannot be supported after say ten years, then } it a disposable item if there are no other service resources. } The dearth of schematics is a huge issue with me since I cannot } talk to the service folks with knowledge and also impacts the } future cut off of service. One is basically buying a $500K } boat anchor that needs replacing every ten year or so. Yep, } sounds like a boat problem. } } Recall that the Amray and early JEOL tools were relatively } easy to fix... electronics-wise. These newer systems are not } at all easy to fix or troubleshoot. } } I'm just blowing the whistle on what I think is a pending } huge problem. } } Alternate opinions? } } gary g. } } } } ==============================Original Headers============================== } 11, 17 -- From gary-at-gaugler.com Fri Aug 18 23:06:32 2006 } 11, 17 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) } 11, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7J46Vmm017237 } 11, 17 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug 2006 23:06:31 -0500 } 11, 17 -- Received: (qmail 17545 invoked from network); 18 Aug 2006 21:06:31 -0700 } 11, 17 -- Received: by simscan 1.1.0 ppid: 17542, pid: 17543, t: 0.1494s } 11, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 } 11, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) } 11, 17 -- by qsmtp2 with SMTP; 18 Aug 2006 21:06:30 -0700 } 11, 17 -- Message-Id: {7.0.1.0.2.20060818204828.0247a458-at-gaugler.com} } 11, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } 11, 17 -- Date: Fri, 18 Aug 2006 21:06:33 -0700 } 11, 17 -- To: MSA listserver {microscopy-at-microscopy.com} } 11, 17 -- From: Gary Gaugler {gary-at-gaugler.com} } 11, 17 -- Subject: New subject-service life of new SEMs } 11, 17 -- Mime-Version: 1.0 } 11, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 6, 20 -- From jfactor-at-ns.purchase.edu Sat Aug 19 16:53:27 2006 6, 20 -- Received: from mta1.srv.hcvlny.cv.net (mta1.srv.hcvlny.cv.net [167.206.4.196]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7JLrRoe012401 6, 20 -- for {microscopy-at-msa.microscopy.com} ; Sat, 19 Aug 2006 16:53:27 -0500 6, 20 -- Received: from [127.0.0.1] (ool-4357ef27.dyn.optonline.net [67.87.239.39]) 6, 20 -- by mta1.srv.hcvlny.cv.net 6, 20 -- (Sun Java System Messaging Server 6.2-6.01 (built Apr 3 2006)) 6, 20 -- with ESMTP id {0J4900GR5M6U8X40-at-mta1.srv.hcvlny.cv.net} for 6, 20 -- microscopy-at-msa.microscopy.com; Sat, 19 Aug 2006 17:54:35 -0400 (EDT) 6, 20 -- Date: Sat, 19 Aug 2006 17:53:39 -0400 6, 20 -- From: Jan Factor {jfactor-at-ns.purchase.edu} 6, 20 -- Subject: Re: [Microscopy] New subject-service life of new SEMs 6, 20 -- In-reply-to: {200608190407.k7J47bjo018742-at-ns.microscopy.com} 6, 20 -- To: gary-at-gaugler.com, Microscopy Listserver {microscopy-at-msa.microscopy.com} 6, 20 -- Message-id: {44E78863.4030605-at-ns.purchase.edu} 6, 20 -- MIME-version: 1.0 6, 20 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 20 -- Content-transfer-encoding: 7BIT 6, 20 -- References: {200608190407.k7J47bjo018742-at-ns.microscopy.com} 6, 20 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) ==============================End of - Headers==============================
I agree as well. The problem is that the older systems were easier to maintain in major part due to the way they were made and the technology at that time--multiple pull out PC cards, socketed ICs, etc. That is all going away now as the makers use TSOP (small outline surface mount IC) ICs, chip resistors and capacitors and pack more functions (FPGAs) onto a single PC board. Result....fewer but more dense PC boards that are probably close to impossible or at best, difficult to repair. However, the other side of this more hands-off manufacturing is greater consistency. This results in fewer failures. The other detrimental factor though is the out-sourcing of portions of the system. The SEM makers build the frame, column and associated parts but have some other company do the electronics or portions of the electronics and/or software. In many cases, the field service folks do not have schematics that accurately reflect the machine they are working on--not to mention that they can no longer replace individual components. It is a replace the whole thing scenario now. The advent of advanced computer control seems to be to keep fixing bugs and adding new or different features to justify the software engineering department or source. Some of these software updates fix things that are so stupid and obvious it seems that software debugging is the job of the customer. That is very bad.
The Devil is in the details. The Devil here is dollars. If the systems are more reliable, people can keep them running for longer periods of time. Remember that most companies have two major cost centers--sales and service. So the sales side sells a new system which is then covered under a maintenance contract by the service side. With a reliable system, the service side makes a nice profit since there are fewer service calls than before. In the mean time, the sales side complains that they are not selling enough systems. The solution? Stop supporting fielded systems to force users to buy a new SEM.
Since I have no financial interest any more with KLA-Tencor I can use them as a good example of how I perceive the internal thinking (or lack thereof) is seen in action. I owned multiple Amray SEMs over the years and at first had Amray contracts. Then, when KLA bought out Amray, service moved to KLA. Over time, KLA began to drop service contracts for their thermionic SEMs. Since my last Amray SEM was an FE, I did not get dropped. Their support prices were quite reasonable compared to today's FESEM maintenance cost. Anyway, as they began dropping thermionic support they also started to lay off service personnel. An obvious cost cutting mentality. Then, as more Amray FE optics were put into KLA inspection tools, the Amray techs got pulled (sucked) into the tool side. Then, behold-- less support for Amray FE SEMs and less support for KLA tools since the techs left leaving a support vacuum. This was in 2003-2004. I suspect that since then, they have either stopped supporting FESEMs or soon will. That is too bad since the later systems were real work horses. I had very few problems with any of my Amray systems. But eventually, the performance just was not there. This was because Amray stopped developing new systems. Software development also stopped (it always was out-sourced). The current manufacturers are not in this non-developmental mode at all.
So the business model of continuous introduction of new models of SEMs and other systems is to "have a growth path" when in fact, earlier models do the intended job just fine, thank you. But they do need support from time to time. Agreed....some need a lot more support than others. But are they newer or older systems? Some are newer or newest, unfortunately. So the service side complains and the reaction from corporate is to cull their product line to simplify the field complement of systems. Optimal in their view is probably that every installation has the exact same system. This would reduce parts logistics, documentation deviations, software option switches, etc. Of course it would. But not all customers' needs are identical.
My gut feeling is--don't count on more than 7-10 years of support from the OEM. And there probably won't be second or third party options. Like I said initially--you are going to be stuck with a boat anchor. It is just a matter of time. This is true for older systems. But I think the time frame to become an anchor is getting shorter. I'm not sure that my crystal ball is any better than any one else's. But I came away from M&M 2006 with this gnawing uncomfortable feeling. Newer isn't always better or best. But in a competitive world, not producing newer may be fatal. If the OEM manufacturers don't survive, one certainly cannot get a service contract from them! It is a horrible business situation.
When buying a new system, you cannot count on determining how long they will support the system. The sales side will tell you whatever you want to hear. The only viable option IMO is to find out what and how old of systems they are currently supporting. Are the postings on this list for SEM/TEM support due to cost reduction at the customer side or lack of support options from the manufacturers? I don't know. That is a good question to ask.
It just seems to me that what has been rather common in the past is pretty quickly being obsoleted. This pertains to how systems are supported, for how long and by whom. But the business model seems to be the same. The issue or point of equipment saturation is quite valid. If all auto makers made cars that lasted 30 years, then who would be the volume new car buyers? As long as the car gets you from point A to point B and back, what is the advantage of a new car? The discriminator is gas mileage, features, etc. With SEM/TEM it is performance. As long as they do what is needed, a newer model might be nice but is not necessary to get the job done.
gary g.
At 02:53 PM 8/19/2006, you wrote: } You are right when you say that these are not disposable items. If } manufacturers have the attitude that we in the academic world can } simply conjure up $250-500,000 because they determine that the } lifespan of their EM is 10 years, then the EM community collectively } should send a strong message to the contrary. My attitude is this: } if an EM manufacturer will not continue to support and keep my EM } going until I am ready (and able) to replace it, then (when I do } find the funds for a } replacement) my next microscope will definitely not be one of } theirs. There are companies that apparently stand up for their } microscopes for the extended lifetimes they should be proud of. As } an example, the lab I was trained in still has the microscope that I } was trained on some 30 years ago (they added a digital camera, and } I'm told it's one of the favored TEMs in the lab). } --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 } --------------------------------------- } } } } gary-at-gaugler.com wrote: } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 13, 21 -- From gary-at-gaugler.com Sat Aug 19 19:36:32 2006 13, 21 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7K0aWeh024692 13, 21 -- for {microscopy-at-microscopy.com} ; Sat, 19 Aug 2006 19:36:32 -0500 13, 21 -- Received: (qmail 31522 invoked from network); 19 Aug 2006 17:36:30 -0700 13, 21 -- Received: by simscan 1.1.0 ppid: 31519, pid: 31520, t: 0.2869s 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 qsmtp1 with SMTP; 19 Aug 2006 17:36:30 -0700 13, 21 -- Message-Id: {7.0.1.0.2.20060819154105.02544cd0-at-gaugler.com} 13, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 21 -- Date: Sat, 19 Aug 2006 17:36:33 -0700 13, 21 -- To: Jan Factor {jfactor-at-ns.purchase.edu} 13, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 21 -- Subject: Re: [Microscopy] New subject-service life of new SEMs 13, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 21 -- In-Reply-To: {44E78863.4030605-at-ns.purchase.edu} 13, 21 -- References: {200608190407.k7J47bjo018742-at-ns.microscopy.com} 13, 21 -- {44E78863.4030605-at-ns.purchase.edu} 13, 21 -- Mime-Version: 1.0 13, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Hi all, I have a friend who needs to make a montage of a large number of optical micrographs. He has about 500 images. Can anybody suggest a PC software package that is easy to use? He is currently using Photoshop. Freeware or something that can be paid for and downloaded would be especially useful. Thanks for any suggestions,
Jennifer Ray Sloppy Materials Science & Engineering, Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 3, 17 -- From jds451-at-psu.edu Sun Aug 20 09:51:57 2006 3, 17 -- Received: from webmail19.cac.psu.edu (webmail19.cac.psu.edu [128.118.1.205]) 3, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7KEpvZI004541 3, 17 -- for {microscopy-at-microscopy.com} ; Sun, 20 Aug 2006 09:51:57 -0500 3, 17 -- Received: (from webmail-at-localhost) 3, 17 -- by webmail19.cac.psu.edu (8.9.3/8.9.0) id KAA20369; 3, 17 -- Sun, 20 Aug 2006 10:51:56 -0400 (EDT) 3, 17 -- Date: Sun, 20 Aug 2006 10:51:56 -0400 (EDT) 3, 17 -- Message-Id: {200608201451.KAA20369-at-webmail19.cac.psu.edu} 3, 17 -- To: microscopy-at-microscopy.com 3, 17 -- Subject: Software: Montage 3, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} 3, 17 -- X-Mailer: Penn State WebMail 3, 17 -- X-Sender: jds451 3, 17 -- X-Originating-IP: 207.255.156.167 3, 17 -- MIME-Version: 1.0 3, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
I have used Panavue ImageAssembler (http://www.panavue.com/) with some success. You do have to load the images in row order. I've sometimes had to stitch the rows first then assemble the rows into the final image rather than assembling the whole image in one pass. ImageAssembler can use either automatic or manual registration for the stitching. The auto registration works reasonably well for image sets with reasonable overlap (20-25%) in the images and if the features are easily distinguishable and not too complex. (I suspect that it just does a cross-correlation.) It will do some image warping and brightness/contrast adjustment to better blend the photos in the stitching process.
The software is pretty easy to use and the price is reasonable ($64 for the standard edition and $129 for the professional edition). The professional version will handle larger images.
The software has worked for me. Your mileage may vary!
Usual disclaimer: I have no vested interest in the product or company.
Good Luck, Henk Colijn
At 10:55 AM 8/20/2006, jds451-at-psu.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
==============================Original Headers============================== 13, 26 -- From colijn.1-at-osu.edu Sun Aug 20 20:19:13 2006 13, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 13, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7L1JClO007521 13, 26 -- for {microscopy-at-microscopy.com} ; Sun, 20 Aug 2006 20:19:13 -0500 13, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 13, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 13, 26 -- id {01M689NUXPKGA8NPYQ-at-er6s1.eng.ohio-state.edu} for 13, 26 -- microscopy-at-microscopy.com; Sun, 20 Aug 2006 21:19:10 -0400 (EDT) 13, 26 -- Received: from HOC3.osu.edu 13, 26 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 13, 26 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 13, 26 -- with ESMTPA id {01M689NU56XYA8MMTU-at-er6s1.eng.ohio-state.edu} ; Sun, 13, 26 -- 20 Aug 2006 21:19:10 -0400 (EDT) 13, 26 -- Date: Sun, 20 Aug 2006 21:18:00 -0400 13, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 13, 26 -- Subject: Re: [Microscopy] Software: Montage 13, 26 -- In-reply-to: {200608201455.k7KEtawS007511-at-ns.microscopy.com} 13, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 13, 26 -- To: jds451-at-psu.edu 13, 26 -- Cc: microscopy-at-microscopy.com 13, 26 -- Message-id: {7.0.1.0.2.20060820205737.022fe018-at-osu.edu} 13, 26 -- MIME-version: 1.0 13, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 13, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 13, 26 -- References: {200608201455.k7KEtawS007511-at-ns.microscopy.com} ==============================End of - Headers==============================
As you may know we run maintenance courses for all types of SEM and TEM and whilst I agree with most of what has been said may I add a point that may help those who are into the first few years of an instrument?
Today you may have PCs, memory, mice, keyboards, printers that are the same as your "new" microscope. In a few years or even weeks the PC you have will become obsolete - DO NOT THROW IT AWAY! In a few years time you may need parts for your microscope that will no longer be available - PCs, memory, mice, keyboards, printers - see what I mean?
Good luck enjoy your new machine, but think ahead!
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: {gary-at-gaugler.com} To: {protrain-at-emcourses.com} Sent: Sunday, August 20, 2006 1:37 AM
==============================Original Headers============================== 9, 33 -- From protrain-at-emcourses.com Mon Aug 21 07:24:34 2006 9, 33 -- Received: from rd01.mail.x-isp.net (rd01.mail.x-isp.net [213.253.171.197]) 9, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7LCOUlb027469 9, 33 -- for {microscopy-at-microscopy.com} ; Mon, 21 Aug 2006 07:24:33 -0500 9, 33 -- Received: from [62.69.65.166] (helo=smtp-c.mail.legend.co.uk) 9, 33 -- by rd01.mail.x-isp.net with smtp (Exim 4.60) 9, 33 -- (envelope-from {protrain-at-emcourses.com} ) 9, 33 -- id 1GF8pS-0005nc-0D 9, 33 -- for microscopy-at-microscopy.com; Mon, 21 Aug 2006 13:24:30 +0100 9, 33 -- Received: (qmail 24839 invoked from network); 21 Aug 2006 12:24:23 -0000 9, 33 -- Received: from unknown (HELO advent) (212.248.148.142) 9, 33 -- by 0 with SMTP; 21 Aug 2006 12:24:23 -0000 9, 33 -- Message-ID: {00db01c6c51c$f5f66680$0300a8c0-at-advent} 9, 33 -- Reply-To: "Steve Chapman" {protrain-at-emcourses.com} 9, 33 -- From: "Steve Chapman" {protrain-at-emcourses.com} 9, 33 -- To: {gary-at-gaugler.com} 9, 33 -- Cc: "American Soc" {microscopy-at-microscopy.com} 9, 33 -- References: {200608200037.k7K0bo8e026515-at-ns.microscopy.com} 9, 33 -- Subject: Re: [Microscopy] Re: New subject-service life of new SEMs 9, 33 -- Date: Mon, 21 Aug 2006 12:05:08 +0100 9, 33 -- Organization: Protrain 9, 33 -- MIME-Version: 1.0 9, 33 -- Content-Type: text/plain; 9, 33 -- format=flowed; 9, 33 -- charset="iso-8859-1"; 9, 33 -- reply-type=original 9, 33 -- Content-Transfer-Encoding: 7bit 9, 33 -- X-Priority: 3 9, 33 -- X-MSMail-Priority: Normal 9, 33 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2527 9, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2527 9, 33 -- X-tmReceived: from [62.69.65.166] (helo=smtp-c.mail.legend.co.uk) 9, 33 -- by rd01.mail.x-isp.net with smtp id 1GF8pS-0005nc-0D;Mon, 21 Aug 2006 13:24:30 +0100 ==============================End of - Headers==============================
Something sound wrong. We have a D50, a D200, and an FE these all use the standard nikon F-mount, and we have routinely switched between these without any problems. The D70 also has an F-mount, I have not tried it specifically but it should be a straight mount. You should NOT need a T-mount adapter (unless you actually have F-mount to T-mount adapters on your FE2 and FTN already), and your microscope phototube mount is a T-mount.
Now, what you will find is that the D70 (like the D50, D80, and D100), will only work in manual exposure mode, as it will treat the microscope mount as a Non-CPU lens.
Now for the D50 and D200, what we have are f-mounts on our phototubes (off of Nikon and Olympus scopes), 2X or 2.5X photo- eye pieces work well, and Nikon right-angle viewfinders. We use the cameras in tethered mode: AC-adapters, USB connections to computers, and run the Nikon Capture 4 software. The nikon software runs the camera directly from the computer (exposure, download, and shutter release). The software is not cheap (Capture 4 is $99, Capture NX is $149). The alternative is an electronic cable release, about $25, (You do NOT need the $150 Nikon remote release) but you still need to down load the images from the memory card.
Now a warning: The D50 has signifcant vibration due to the mirror, and like the D70, it does not have a mirror lock-up feature. When using the D50 on our Nikon Optiphot (very similar to the Diaphot) the vibrations are such that the images with 60X and 100X objectives are not useable, 40X is poor but useable. I do not know about the D70's vibration. We just got through testing the D200, and it inherently has much much lower vibration than the D50, and secondly it does have a "Mirror up, shutter delay".
Good luck.
On 18 Aug 2006, at 18:06, KLRadke-at-ucdavis.edu wrote:
} } Email: KLRadke-at-ucdavis.edu } Name: Kathryn Radke } } Organization: Univ. California, Davis } } Title-Subject: [Filtered] LM - coupling digital camera back to Nikon } Diaphot } } Question: I'd like to use my Nikon D70s camera back on my lab's 1985 } Nikon Diaphot inverted cell culture microscope. The Diaphot front port } has a fitting that couples to Nikon film backs (e.g. FE2, FTN) having } Nikon F bayonet type lens mount. When the microscope is set up } properly, the image viewed through the eyepieces focused on the film. } } The D70S back won't fit directly onto the Diaphot. A local camera } store tells me that I could use a T mount adapter that is designed to } couple a digital back with a telescope. With the T mount adapter in } place, the digital back can be used in Manual mode and the shutter } speed can be set using the "command dial" on the camera back. I would } also need to purchase a remote cord to operate the back without } shaking the microscope. } } I would be most grateful if anyone can give some feedback about } necessary parts, operation, etc. Are there websites that have } relevant information? } } Thank you. } Kathryn } } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== 9, 12 -- From } zaluzec-at-microscopy.com Fri Aug 18 18:05:52 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 } k7IN5pPG017456 9, 12 -- for {microscopy-at-microscopy.com} ; Fri, 18 Aug } 2006 18:05:52 -0500 9, 12 -- Mime-Version: 1.0 9, 12 -- X-Sender: } (Unverified) 9, 12 -- Message-Id: } {p06110401c10bf8404212-at-[206.69.208.22]} 9, 12 -- Date: Fri, 18 Aug } 2006 18:05:50 -0500 9, 12 -- To: microscopy-at-microscopy.com 9, 12 -- } From: KLRadke-at-ucdavis.edu (by way of MicroscopyListserver) 9, 12 -- } Subject: viaWWW: LM - coupling digital camera back to Nikon Diaphot 9, } 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers==============================
Richard E. Edelmann, Ph.D. EXPO Editor, 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============================== 13, 25 -- From edelmare-at-muohio.edu Mon Aug 21 08:14:58 2006 13, 25 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 13, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7LDEwxr006149 13, 25 -- for {microscopy-at-Microscopy.com} ; Mon, 21 Aug 2006 08:14:58 -0500 13, 25 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 13, 25 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7LDEuwv002611; 13, 25 -- Mon, 21 Aug 2006 09:14:56 -0400 13, 25 -- Received: from emf03 ([134.53.14.97]) 13, 25 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7LDEto0008096; 13, 25 -- Mon, 21 Aug 2006 09:14:56 -0400 13, 25 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 13, 25 -- To: KLRadke-at-ucdavis.edu 13, 25 -- Date: Mon, 21 Aug 2006 09:17:00 -0400 13, 25 -- MIME-Version: 1.0 13, 25 -- Content-type: text/plain; charset=US-ASCII 13, 25 -- Content-transfer-encoding: 7BIT 13, 25 -- Subject: Re: [Microscopy] viaWWW: LM - coupling digital camera back to Nikon Diaphot 13, 25 -- CC: microscopy-at-Microscopy.com 13, 25 -- Message-ID: {44E97A0C.28389.DCD8C08-at-localhost} 13, 25 -- Priority: normal 13, 25 -- In-reply-to: {200608182306.k7IN6bYr020112-at-ns.microscopy.com} 13, 25 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 13, 25 -- X-Real-ConnectIP: 134.53.14.97 13, 25 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.67 13, 25 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 ==============================End of - Headers==============================
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Email: aleb-at-nhm.ac.uk Name: Alex Ball
Organization: the Natural history Museum, London
Title-Subject: [Filtered] US - based EM suppliers
Question: Dear all,
With the current restrictions on flying with liquids/gels I am trying to get a list of US and European suppliers of EM chemicals, specifically fixatives.
I would be very grateful if you could forward to me details of suppliers who you think might be willing to send out EM grade glutaraldehyde in 10ml aliquots (glass ampoules).
I am aware of EMS and Ted Pella, although I do not have either of their US catalogues (I'll be writing to them formally in the near future).
Also, one of my colleagues spent some time working in a marine lab and was able to source pre-buffered glutaraldehyde, ready mixed to 2.5% concentration and 0.1M buffered to 7.2pH. Of course he forgot to take details of the supplier!!!! Has anyone seen this? I'd love to track this down as it would make supplying fixatives for our field researchers so easy.
Regards and thanks in advance,
Alex Ball E M unit manager, The Natural History Museum, London (Formerly B.M.(N.H.))
The coupling problem is probably caused by a ring around the camera mount attached to the microscope or camera adapter unit. The ring does not interfere with mounting the film cameras designed to mount on the microscope, but prevents mounting of some digital SLR F-mount cameras. The ring, which seems to be entirely cosmetic, is held in place with fine screws and is easily removed. Once the ring is removed the digital camera should lock in place properly.
The problem of camera vibration can be solved by setting the manual time exposure to some large value like 4 seconds and using one of the exposure control units made by Nikon (AFX, UFX etc) to control the exposure. Trigger the camera, wait a second, then trigger the exposure control unit which opens and closes the shutter in the camera adapter unit that was designed for 35mm film cameras. If you don't have one of these adapters and control units, they regularly show up on eBay.
The sensor unit of digital cameras is smaller than 35mm film format (except for the Kodak DCS 14 megapixel cameras) so you should use the 2.5x photo eyepiece rather than the 4x or 5x.
Ralph Common Division of Human Pathology Michigan State University
==============================Original Headers============================== 6, 24 -- From rcommon-at-msu.edu Mon Aug 21 11:07:23 2006 6, 24 -- Received: from sys21.mail.msu.edu (sys21.mail.msu.edu [35.9.75.121]) 6, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7LG7NuX029919 6, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 21 Aug 2006 11:07:23 -0500 6, 24 -- Received: from [35.9.122.125] (helo=emlab) 6, 24 -- by sys21.mail.msu.edu with esmtpsa (Exim 4.52 #1) 6, 24 -- (TLSv1:RC4-MD5:128) 6, 24 -- id 1GFCJ9-00042W-CR 6, 24 -- for Microscopy-at-microscopy.com; Mon, 21 Aug 2006 12:07:23 -0400 6, 24 -- From: "Ralph Common" {rcommon-at-msu.edu} 6, 24 -- To: {Microscopy-at-microscopy.com} 6, 24 -- Subject: LM - coupling digital camera back to Nikon Diaphot 6, 24 -- Date: Mon, 21 Aug 2006 12:08:33 -0400 6, 24 -- Message-ID: {000001c6c53c$0e145140$7d7a0923-at-msu.edu} 6, 24 -- MIME-Version: 1.0 6, 24 -- Content-Type: text/plain; 6, 24 -- charset="iso-8859-1" 6, 24 -- Content-Transfer-Encoding: 7bit 6, 24 -- X-Priority: 3 (Normal) 6, 24 -- X-MSMail-Priority: Normal 6, 24 -- X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2911.0) 6, 24 -- Importance: Normal 6, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 6, 24 -- X-Virus: None found by Clam AV ==============================End of - Headers==============================
The general construction and functioning of cold cathode discharge gauges is discussed and illustrated in Sect. 3.2.2 (p. 99) of my book Vacuum Methods in Electron Microscopy (available from SPI Supplies, Ladd, M. E. Taylor, etc.). In particular, the photo of a disassembled gauge tube shown on page 101 might be helpful in giving you an idea of what to expect when taking your gauge tube apart for cleaning. The cleaning of Penning gauges is a rather common operation, so don't be afraid to undertake it - just be careful, and proceed with caution. In particular, don't get steel wool or any other magnetic cleaning agent in the neighborhood of the magnets that are part of the gauge, or you'll have a VERY difficult time getting the residue of the off.
The cleaning procedure recommended by Valery sounds like a simple one, so give it a try. You might also want to look over the comments about cleaning procedures in general given on page 70 of my book.
Good luck, WCB -- Wilbur C. Bigelow, Professor Emeritus Materials Sci. & Engr., Univ. of Michigan Ann Arbor, Michigan 48109-2136 e-mail: bigelow-at-engin.umich.edu; Fx:734-763-4788; Ph:734-975-0858 Address mail to: 2911 Whittier Court Ann Arbor, MI 48104-6731
==============================Original Headers============================== 3, 14 -- From bigelow-at-engin.umich.edu Mon Aug 21 15:02:45 2006 3, 14 -- Received: from srvr22.engin.umich.edu (srvr22.engin.umich.edu [141.213.75.21]) 3, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7LK2jxg011940 3, 14 -- for {microscopy-at-microscopy.com} ; Mon, 21 Aug 2006 15:02:45 -0500 3, 14 -- Received: from [141.212.131.221] (bigelow-g4.engin.umich.edu [141.212.131.221]) 3, 14 -- by srvr22.engin.umich.edu (8.13.6/8.13.6) with ESMTP id k7LK2iYV015998 3, 14 -- for {microscopy-at-microscopy.com} ; Mon, 21 Aug 2006 16:02:45 -0400 (EDT) 3, 14 -- Mime-Version: 1.0 3, 14 -- Message-Id: {p06210202c10fbcd1bac3-at-[141.212.131.221]} 3, 14 -- Date: Mon, 21 Aug 2006 16:02:43 -0400 3, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 3, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 3, 14 -- Subject: [Microscopy] RE: Cleaning Penning gauges 3, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
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Email: a.ball-at-nhm.ac.uk Name: Alex Ball
Organization: Natural history Museum, London
Title-Subject: [Filtered] US - based EM suppliers - revised question
Question: Dear all,
Earlier today i posted a question about EM fixatives and i have had several useful replies. However, I don't think I was clear about why I wanted details of US suppliers.
Many of the researchers from the Natural History Museum conduct fieldwork overseas where they collect and fix specimens in the field or within other laboratories. In the past we supplied fixatives and buffers for them to take with them. The regulations over transport of small amounts of fixatives were sufficiently fuzzy for this type of material to be carried aboard aircraft without question.
Our post-911 world means this is no longer practical, so I was hoping to gather a list of useful contacts or suppliers so that researchers flying into the continental USA, including South America might be able to arrange for delivery of fixatives to them in order to still be able to fix material in the field.
Several people have sent me details of UK-based suppliers, but it's the the US-based ones that I really need to make contact with.
Sorry for the confusion and thanks for the leads sent to me so far,
Regards,
Alex
Dr Alex Ball Electron Microscope Unit Manager The Natural History Museum London SW7 9BD
Greetings, Can anyone tell me a reliable method to stain starch grains in sections of plant tissue embedded in Spurr's resin? Also useful to know whether polarized light would be suitable. I hope this is easy and I get lots of replies!
As ever, Tobias -- _ ____ __ ____ / \ / / \ / \ \ Tobias I. Baskin / / / / \ \ \ Biology Department /_ / __ /__ \ \ \__ 611 N. Pleasant St. / / / \ \ \ University of Massachusetts / / / \ \ \ Amherst, MA, 01003 / / ___ / \ \__/ \ ____ http://www.bio.umass.edu/biology/baskin/ Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
==============================Original Headers============================== 2, 16 -- From baskin-at-bio.umass.edu Tue Aug 22 09:20:42 2006 2, 16 -- Received: from marlin.bio.umass.edu (marlin.bio.umass.edu [128.119.55.19]) 2, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MEKgaO016331 2, 16 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 09:20:42 -0500 2, 16 -- Received: from [172.30.55.79] (eutopia [128.119.55.30]) 2, 16 -- by marlin.bio.umass.edu (8.13.6/8.13.6) with ESMTP id k7MEKcMV012516 2, 16 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:20:39 -0400 (EDT) 2, 16 -- Mime-Version: 1.0 2, 16 -- Message-Id: {p06230906c110c25e16f3-at-[172.30.55.79]} 2, 16 -- Date: Tue, 22 Aug 2006 10:20:38 -0400 2, 16 -- To: microscopy-at-microscopy.com 2, 16 -- From: Tobias Baskin {baskin-at-bio.umass.edu} 2, 16 -- Subject: staining starch in sections 2, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 2, 16 -- X-Greylist: Sender succeded SMTP AUTH authentication, not delayed by milter-greylist-1.6 (marlin.bio.umass.edu [128.119.55.19]); Tue, 22 Aug 2006 10:20:39 -0400 (EDT) 2, 16 -- X-Scanned-By: MIMEDefang 2.54 on 128.119.55.19 ==============================End of - Headers==============================
I think I have an easy solution for a common problem and I just wanted to share it. I am sorry if I bring nothing new, but perhaps it really helps. At least it cannot harm :-)
When working with my TEMcam Megaview III and the software AnalySIS (no personal interest blablabla...) the pictures are saved in 16bit TIFF format. This is a problem when you want to work with them because this format is not recognized by most image processing software (This problem is so common that it appears in the FAQ of AnalySIS). However when this format is loaded in Adobe Photoshop (no personal....) the picture comes completely dark. A rather simple solution to recover your picture is to use the "auto contrast" and/or "auto level" option and your nice picture appears in all its splendor right before your eyes!
Hope this was helpful. Have a nice day (for the american), a nice sleep (for the australian) or a nice evening (for the european).
Stephane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Tue Aug 22 10:22:55 2006 7, 18 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.91.142]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7MFMtJ5027602 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:22:55 -0500 7, 18 -- Received: (qmail 21837 invoked by uid 60001); 22 Aug 2006 15:22:55 -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=FP2WVswviXCCARHnXwNLh4HVu9wy6B+uLcz1srxmBXfOK7AOK1yXsSzEW/ImrlWN4trgP7WdCahgLGDn8k2mb/DEqR/6OtgyruYcPY4CSc5LB0WGkWkbw38+DvR5OuvAvRI51DYVL4elyoXJJaPVjoBn3cOnsVMP+LHoCAm6+yA= ; 7, 18 -- Message-ID: {20060822152255.21835.qmail-at-web37410.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Tue, 22 Aug 2006 08:22:55 PDT 7, 18 -- Date: Tue, 22 Aug 2006 08:22:55 -0700 (PDT) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: 16bit TIFF images in Photoshop 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==============================
Okay, 99% of all my ultra-microtomy has been done with epoxy/resin embedded 'biological' samples.
Recently I was asked about cutting a thin bi-metalic film on the surface of a silicon wafer.
My initial gut reaction to this was "cut a Si wafer in cross-section with one of our diamond knives? Are you kidding?"
For those out there more immersed in the materials side of TEM prep, can a Si wafer be cut in cross section with a regular 45º Ultra Diamond knife? Would thinning the wafer down prior to deposition make it easier to cut? Or easier on the knife? If possible to cut, would these sections just be floated off in water like standard samples? Or best to leave dry and manipulate with eye-lashes or Dalmatian hair.
Thank you for bearing with these questions on materials prep from a biological based pov. And no, this is the only way to get this specific film thin enough (cutting), Ion Mill, polishing, no other 'typical' thinning prep works.
Geoff Williams Leduc Bioimaging Facility Manager Brown University
16 bit TIFFs are a fairly common format for scientific image data, and there are a variety of programs intended for scientific use which support them. In addition to many commercial products both cheaper and more expensive than Photoshop (DigitalMicrograph, IDL, Igor Pro, etc. etc.) there is an excellent, free program called ImageJ available from
{http://rsb.info.nih.gov/ij/}
It reads and displays the 16 bit TIFF images from our MegaVIEW / AnalySIS system with no additional steps.
There is also a potentially serious drawback to the Photoshop procedure you suggest. Photoshop discards the original grey-scale values of the image when you use "Auto Levels", which breaks the quantitative connection between the grey-scale value in the image and the absolute number of electrons that struck the detector. This will often make quantitative analysis of the electron image or diffraction pattern impossible and makes it difficult to even judge the relative average brightness of two different images to guess, for example, the sample thickness.
I apologize for the long reply, but this is one of my pet peeves: what comes off the microscope has the potential to be quantitative data, not just pictures. Software developed for photographs and desktop publishing is not generally the best choice for dealing with scientific data.
Best wishes, Paul Voyles
Paul Voyles Materials Science and Engineering University of Wisconsin, Madison 1509 University Ave, Rm 223 Madison, WI 53706-1595 voice: (608) 265-6740 fax: (608) 262-8353 voyles-at-engr.wisc.edu http://tem.msae.wisc.edu
nizets2-at-yahoo.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear listers, } } I think I have an easy solution for a common problem } and I just wanted to share it. I am sorry if I bring } nothing new, but perhaps it really helps. At least it } cannot harm :-) } } When working with my TEMcam Megaview III and the } software AnalySIS (no personal interest blablabla...) } the pictures are saved in 16bit TIFF format. This is a } problem when you want to work with them because this } format is not recognized by most image processing } software (This problem is so common that it appears in } the FAQ of AnalySIS). However when this format is } loaded in Adobe Photoshop (no personal....) the } picture comes completely dark. A rather simple } solution to recover your picture is to use the "auto } contrast" and/or "auto level" option and your nice } picture appears in all its splendor right before your } eyes! } } Hope this was helpful. } Have a nice day (for the american), a nice sleep (for } the australian) or a nice evening (for the european). } } Stephane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com
==============================Original Headers============================== 11, 25 -- From voyles-at-engr.wisc.edu Tue Aug 22 10:45:08 2006 11, 25 -- Received: from mail.cae.wisc.edu (mail.cae.wisc.edu [144.92.13.31]) 11, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MFj8iP016024 11, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:45:08 -0500 11, 25 -- Received: from portkey.cae.wisc.edu (portkey.cae.wisc.edu [144.92.13.118]) 11, 25 -- by mail.cae.wisc.edu (8.13.3+Sun/8.13.4) with ESMTP id k7MFiwKs006707 11, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:44:58 -0500 (CDT) 11, 25 -- Received: from [128.104.200.57] (voylespad1.msae.wisc.edu [128.104.200.57]) 11, 25 -- (authenticated bits=0) 11, 25 -- by portkey.cae.wisc.edu (8.13.4/8.13.4/Debian-3sarge1) with ESMTP id k7MFiwoJ032645 11, 25 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 11, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:44:58 -0500 11, 25 -- Message-ID: {44EB2674.5020705-at-engr.wisc.edu} 11, 25 -- Date: Tue, 22 Aug 2006 10:44:52 -0500 11, 25 -- From: Paul Voyles {voyles-at-engr.wisc.edu} 11, 25 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 11, 25 -- MIME-Version: 1.0 11, 25 -- To: Microscopy-at-microscopy.com 11, 25 -- Subject: Re: [Microscopy] 16bit TIFF images in Photoshop 11, 25 -- References: {200608221524.k7MFO45e029784-at-ns.microscopy.com} 11, 25 -- In-Reply-To: {200608221524.k7MFO45e029784-at-ns.microscopy.com} 11, 25 -- Content-Type: text/plain; charset=ISO-8859-15; format=flowed 11, 25 -- Content-Transfer-Encoding: 7bit 11, 25 -- X-CAE-MailScanner-Information: Please contact security-at-engr.wisc.edu if this message contains a virus or has been corrupted in delivery. 11, 25 -- X-CAE-MailScanner: Found to be clean (benji) ==============================End of - Headers==============================
I do not know exactly what I can divulge, but they have tried tripod polishing, and ion-milling but I am told they elevate the temperature and the layers need to be prepped and kept at room temps until imaging.
Does that say enough? I still feel it's a bit irrelevant to the initial question.
Is it possible to section a Si wafer? Am I correct in being skeptical? Or should I just kill a $4k knife or try an ancient already messed up and questionably anchored diamond in the attempt?
I just figured someone out there might be able to share a bit of practical experience.
Thanks!
Geoff Williams Leduc Bioimaging Facility Manager Brown University
Try the freeware Autostitch ( http://www.autostitch.net ). It is free, it works very well even if the interface of the demo version is crude and basic.
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Jennifer,
I have used Panavue ImageAssembler (http://www.panavue.com/) with some success. You do have to load the images in row order. I've sometimes had to stitch the rows first then assemble the rows into the final image rather than assembling the whole image in one pass. ImageAssembler can use either automatic or manual registration for the stitching. The auto registration works reasonably well for image sets with reasonable overlap (20-25%) in the images and if the features are easily distinguishable and not too complex. (I suspect that it just does a cross-correlation.) It will do some image warping and brightness/contrast adjustment to better blend the photos in the stitching process.
The software is pretty easy to use and the price is reasonable ($64 for the standard edition and $129 for the professional edition). The professional version will handle larger images.
The software has worked for me. Your mileage may vary!
Usual disclaimer: I have no vested interest in the product or company.
Good Luck, Henk Colijn
At 10:55 AM 8/20/2006, jds451-at-psu.edu wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
==============================Original Headers============================== 13, 26 -- From colijn.1-at-osu.edu Sun Aug 20 20:19:13 2006 13, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 13, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7L1JClO007521 13, 26 -- for {microscopy-at-microscopy.com} ; Sun, 20 Aug 2006 20:19:13 -0500 13, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 13, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 13, 26 -- id {01M689NUXPKGA8NPYQ-at-er6s1.eng.ohio-state.edu} for 13, 26 -- microscopy-at-microscopy.com; Sun, 20 Aug 2006 21:19:10 -0400 (EDT) 13, 26 -- Received: from HOC3.osu.edu 13, 26 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 13, 26 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 13, 26 -- with ESMTPA id {01M689NU56XYA8MMTU-at-er6s1.eng.ohio-state.edu} ; Sun, 13, 26 -- 20 Aug 2006 21:19:10 -0400 (EDT) 13, 26 -- Date: Sun, 20 Aug 2006 21:18:00 -0400 13, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 13, 26 -- Subject: Re: [Microscopy] Software: Montage 13, 26 -- In-reply-to: {200608201455.k7KEtawS007511-at-ns.microscopy.com} 13, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 13, 26 -- To: jds451-at-psu.edu 13, 26 -- Cc: microscopy-at-microscopy.com 13, 26 -- Message-id: {7.0.1.0.2.20060820205737.022fe018-at-osu.edu} 13, 26 -- MIME-version: 1.0 13, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 13, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 13, 26 -- References: {200608201455.k7KEtawS007511-at-ns.microscopy.com} ==============================End of - Headers==============================
==============================Original Headers============================== 24, 26 -- From jean-paul.bailon-at-polymtl.ca Tue Aug 22 11:04:57 2006 24, 26 -- Received: from smtp.polymtl.ca (smtp.polymtl.ca [132.207.4.11]) 24, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MG4vVr002519 24, 26 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 11:04:57 -0500 24, 26 -- Received: from jpbailon (jmdorlot.metal.polymtl.ca [132.207.53.29]) 24, 26 -- by smtp.polymtl.ca (8.13.6/8.13.6) with ESMTP id k7MG4qdA017147 24, 26 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 12:04:56 -0400 24, 26 -- Return-Receipt-To: "Jean-Paul Bailon" {jean-paul.bailon-at-polymtl.ca} 24, 26 -- Reply-To: {jean-paul.bailon-at-polymtl.ca} 24, 26 -- From: "Jean-Paul Bailon" {jean-paul.bailon-at-polymtl.ca} 24, 26 -- To: {Microscopy-at-microscopy.com} 24, 26 -- Subject: RE: [Microscopy] Re: Software: Montage 24, 26 -- Date: Tue, 22 Aug 2006 12:05:05 -0400 24, 26 -- Organization: Ecole Polytechnique de Montreal 24, 26 -- Message-ID: {!&!AAAAAAAAAAAYAAAAAAAAAIv5ENKzUdMRv4cAUAR8QvvCgAAAEAAAAPPNakJfCY9HnJXfsHX2hyABAAAAAA==-at-polymtl.ca} 24, 26 -- MIME-Version: 1.0 24, 26 -- Content-Type: text/plain; 24, 26 -- charset="iso-8859-1" 24, 26 -- X-Mailer: Microsoft Office Outlook 11 24, 26 -- In-Reply-To: {200608210120.k7L1KdsH009154-at-ns.microscopy.com} 24, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 24, 26 -- Thread-Index: AcbEwAaXEu6ez8u1S7G5U6mzqzR6eABQ9fJw 24, 26 -- Disposition-Notification-To: "Jean-Paul Bailon" {jean-paul.bailon-at-polymtl.ca} 24, 26 -- X-Poly-FromMTA: (jmdorlot.metal.polymtl.ca [132.207.53.29]) at Tue, 22 Aug 2006 16:04:52 +0000 24, 26 -- Content-Transfer-Encoding: 8bit 24, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7MG4vVr002519 ==============================End of - Headers==============================
Hi Geoffrey, A similar question came up a while ago about samples getting too hot during prep or in the TEM itself. I've managed to prepare Au:Ge and Pd:Zn:Au metal layers on III-Vs using fairly conventional techniques; these metals interdiffuse below 100C, so unless you have to keep the temperature really low for your samples it should be possible. The only difference to my standard protocol is to use superglue to mount the sample rather than thermoplastic wax, and soak the sample off in acetone (which I think is the standard practice for tripod polishing anyway?). Since I work with III-Vs like InP I always have to keep the energy going into the sample to a minimum during ion miling, but I find I can hit it with 6 kV Ar+ ions in a PIPS as long as I keep the incidence angle below 3 degrees. However when I was using an ion tech mill which didn't have a modulated beam or go below 10 degrees incidence I had to use LN2 cooling. Contact me off list if you want more details. As for microtomy, I have no idea. I have heard of it being done and read a paper or two, but I haven't got any further than being envious of the idea of making an electron transparent TEM sample in less that 10 mins.. or perhaps that's just my fantasy of how quick microtomy can be..
-----Original Message----- X-from: Geoffrey_Williams-at-brown.edu [mailto:Geoffrey_Williams-at-brown.edu] Sent: 22 August 2006 16:33 To: Richard Beanland
Okay, 99% of all my ultra-microtomy has been done with epoxy/resin embedded 'biological' samples.
Recently I was asked about cutting a thin bi-metalic film on the surface of a silicon wafer.
My initial gut reaction to this was "cut a Si wafer in cross-section with one of our diamond knives? Are you kidding?"
For those out there more immersed in the materials side of TEM prep, can a Si wafer be cut in cross section with a regular 45º Ultra Diamond knife? Would thinning the wafer down prior to deposition make it easier to cut? Or easier on the knife? If possible to cut, would these sections just be floated off in water like standard samples? Or best to leave dry and manipulate with eye-lashes or Dalmatian hair.
Thank you for bearing with these questions on materials prep from a biological based pov. And no, this is the only way to get this specific film thin enough (cutting), Ion Mill, polishing, no other 'typical' thinning prep works.
Geoff Williams Leduc Bioimaging Facility Manager Brown University
==============================Original Headers============================== 8, 29 -- From Geoffrey_Williams-at-brown.edu Tue Aug 22 10:32:17 2006 8, 29 -- Received: from draco.services.brown.edu (draco.services.brown.edu [128.148.106.172]) 8, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MFWG1H005521 8, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:32:17 -0500 8, 29 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 8, 29 -- by draco.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k7MFWGMw023143 8, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 11:32:16 -0400 (EDT) 8, 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); 8, 29 -- Tue, 22 Aug 2006 11:32:16 -0400 8, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 29 -- Content-class: urn:content-classes:message 8, 29 -- MIME-Version: 1.0 8, 29 -- Content-Type: text/plain; 8, 29 -- charset="iso-8859-1" 8, 29 -- Subject: Sectioning Si Wafers 8, 29 -- Date: Tue, 22 Aug 2006 11:32:15 -0400 8, 29 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F047F6786-at-MAIL1.AD.Brown.Edu} 8, 29 -- X-MS-Has-Attach: 8, 29 -- X-MS-TNEF-Correlator: 8, 29 -- Thread-Topic: Sectioning Si Wafers 8, 29 -- Thread-Index: AcbGACVCTVY7vLTfSbKUtWaWeb/iew== 8, 29 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 8, 29 -- To: {Microscopy-at-microscopy.com} 8, 29 -- X-OriginalArrivalTime: 22 Aug 2006 15:32:16.0556 (UTC) FILETIME=[25F216C0:01C6C600] 8, 29 -- X-Brown-Proofpoint: Not Infected 8, 29 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0608080000 definitions=main-0608220002 8, 29 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0608080000 definitions=main-0608220002 8, 29 -- Content-Transfer-Encoding: 8bit 8, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7MFWG1H005521 ==============================End of - Headers==============================
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==============================Original Headers============================== 21, 32 -- From richard.beanland-at-bookham.com Tue Aug 22 11:24:29 2006 21, 32 -- Received: from mail80.messagelabs.com (mail80.messagelabs.com [195.245.230.163]) 21, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7MGOSqH014905 21, 32 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 11:24:29 -0500 21, 32 -- X-VirusChecked: Checked 21, 32 -- X-Env-Sender: richard.beanland-at-bookham.com 21, 32 -- X-Msg-Ref: server-4.tower-80.messagelabs.com!1156263867!1189339!1 21, 32 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 21, 32 -- X-Originating-IP: [213.249.209.179] 21, 32 -- Received: (qmail 7650 invoked from network); 22 Aug 2006 16:24:27 -0000 21, 32 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 21, 32 -- by server-4.tower-80.messagelabs.com with SMTP; 22 Aug 2006 16:24:27 -0000 21, 32 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 21, 32 -- Tue, 22 Aug 2006 17:26:47 +0100 21, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 21, 32 -- Content-class: urn:content-classes:message 21, 32 -- MIME-Version: 1.0 21, 32 -- Content-Type: text/plain; 21, 32 -- charset="iso-8859-1" 21, 32 -- Subject: RE: [Microscopy] Sectioning Si Wafers 21, 32 -- Date: Tue, 22 Aug 2006 17:26:49 +0100 21, 32 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E1BA33A-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 21, 32 -- X-MS-Has-Attach: 21, 32 -- X-MS-TNEF-Correlator: 21, 32 -- Thread-Topic: [Microscopy] Sectioning Si Wafers 21, 32 -- Thread-Index: AcbGAJfjm2P9z+WlQLCh7Ucg5/QC4QABIvnw 21, 32 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 21, 32 -- To: {Geoffrey_Williams-at-brown.edu} 21, 32 -- Cc: {microscopy-at-microscopy.com} 21, 32 -- X-OriginalArrivalTime: 22 Aug 2006 16:26:47.0846 (UTC) FILETIME=[C3C95460:01C6C607] 21, 32 -- Content-Transfer-Encoding: 8bit 21, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7MGOSqH014905 ==============================End of - Headers==============================
Cryo ion-milling is one option, FIB liftout is another. A third option is the small-angle cleavage technique that John McCaffrey and Scott Walck developed. The cleavage technique requires some practice, but if done right the fracture should propagate right through the surface film and give you thin damage free samples without heating.
Scott is now at Southbay Technology (walck-at-southbaytech.com) but monitors the listserver. (Scott, Would you like to add anything?)
Cheers, Henk Colijn
At 12:01 PM 08/22/06, Geoffrey_Williams-at-brown.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.
==============================Original Headers============================== 11, 26 -- From colijn.1-at-osu.edu Tue Aug 22 12:50:09 2006 11, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 11, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MHo91V027091 11, 26 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 12:50:09 -0500 11, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 11, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 11, 26 -- id {01M6AMKSUYR4A8MJ3P-at-er6s1.eng.ohio-state.edu} for 11, 26 -- microscopy-at-microscopy.com; Tue, 22 Aug 2006 13:50:08 -0400 (EDT) 11, 26 -- Received: from HOC1.ecr6.ohio-state.edu 11, 26 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 11, 26 -- (PMDF V6.2-1x11 #31056) 11, 26 -- with ESMTPA id {01M6AMKRYQE0A8PT39-at-er6s1.eng.ohio-state.edu} ; Tue, 11, 26 -- 22 Aug 2006 13:50:07 -0400 (EDT) 11, 26 -- Date: Tue, 22 Aug 2006 13:51:34 -0400 11, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 11, 26 -- Subject: Re: [Microscopy] Sectioning Si, clarification 11, 26 -- In-reply-to: {200608221601.k7MG1qja029254-at-ns.microscopy.com} 11, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 11, 26 -- To: Geoffrey_Williams-at-brown.edu 11, 26 -- Cc: microscopy-at-microscopy.com, walck-at-southbaytech.com 11, 26 -- Message-id: {7.0.1.0.2.20060822134349.03791640-at-osu.edu} 11, 26 -- MIME-version: 1.0 11, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 11, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 11, 26 -- References: {200608221601.k7MG1qja029254-at-ns.microscopy.com} ==============================End of - Headers==============================
I was actually dumb enough to try microtoming thin films (non-metallic) on silicon wafers for awhile. I got the best results (this is a relative term) by first embedding small pieces of the wafer in epoxy (Embed 812), then using a polisher and diamond films to polish away most of the silicon from the back side. I think I wedged it slightly to obtain a red area of silicon . I then re-embedded the whole thing to encapsulate it in epoxy, then block trimmed to get to a nice thin area. You want to be cutting through the least amount of silicon possible. I used fairly slow cutting speeds and a water-filled boat. No problems picking up sections or placing on filmed grids, but the material beats up the knife fast so I kept having to move to another knife area.
As for the TEM results, we did get some useful pictures, but the silicon does not cut per se, it's more like a repeatable fracture. So you just look among the pieces for an intact place, which can take up some scope time just finding a good area. If you really cannot prepare your material with other methods (polishing/ion-milling, FIB), or you are trying to assess sample preparation artifacts from ion energies, then it can be useful. But it is tedious, both on the prep side and in the microscope.
I don't have experience with your type of material, but I would suggest first trying tripod polishing or dimpling, and low-energy/low-angle ion-milling.
Feel free to contact me offline if you would like more details.
Leslie
Leslie Krupp (Thompson) IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 7, 28 -- From lkrupp-at-us.ibm.com Tue Aug 22 12:59:55 2006 7, 28 -- Received: from e2.ny.us.ibm.com (e2.ny.us.ibm.com [32.97.182.142]) 7, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MHxtw5005044 7, 28 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 12:59:55 -0500 7, 28 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 7, 28 -- by e2.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k7MHxoSn013452 7, 28 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 13:59:50 -0400 7, 28 -- Received: from d01av04.pok.ibm.com (d01av04.pok.ibm.com [9.56.224.64]) 7, 28 -- by d01relay02.pok.ibm.com (8.13.6/8.13.6/NCO v8.1.1) with ESMTP id k7MHxoX3294042 7, 28 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 13:59:50 -0400 7, 28 -- Received: from d01av04.pok.ibm.com (loopback [127.0.0.1]) 7, 28 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k7MHxniq026488 7, 28 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 13:59:49 -0400 7, 28 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 7, 28 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k7MHxnur026485 7, 28 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 13:59:49 -0400 7, 28 -- In-Reply-To: {200608221631.k7MGVMQJ022467-at-ns.microscopy.com} 7, 28 -- To: microscopy-at-microscopy.com 7, 28 -- Subject: RE: Sectioning Si Wafers 7, 28 -- MIME-Version: 1.0 7, 28 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 7, 28 -- Message-ID: {OFB5C3E18B.7BA6F5D9-ON852571D2.0062D1E7-882571D2.0062DCF0-at-us.ibm.com} 7, 28 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 7, 28 -- Date: Tue, 22 Aug 2006 10:59:49 -0700 7, 28 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF269 | June 22, 2006) at 7, 28 -- 08/22/2006 13:59:49, 7, 28 -- Serialize complete at 08/22/2006 13:59:49 7, 28 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
No you can't "cut" Si with a diamond knife. Si cleaves very easily. When the knife touches the Si, the Si cleaves along one of its preferred {111} crystallographic cleaving planes. That might be fine if you orient the Si such that a Si {111} crystallographic direction is perpendicular to the block face. Then if you are VERY lucky, you MIGHT get a cleaved piece of Si oriented with {111} faces (at the risk of your knife). Any article that I've seen that claims to have microtomed Si has shown lots of pictures of tiny {111} Si sections.
Assuming that you are successful, so what! There is no, or very little interest, in anything in the semiconductor world or in potentially oriented thin films deposited on Si, that is best displayed on top of a Si cross section with a {111} block face orientation. The angles are wrong.
Si wafers in semiconductor processing are almost all with (or very close to) a Si {001} direction perpendicular to the Si wafer surface. Devices fabricated in, or patterns deposited on, these wafers are not randomly oriented. They are almost always aligned with {011} planes perpendicular to the Si surface, i.e. the x, y orientations of the devices and patterns are parallel to {011} directions in the Si surface. Cross sections of interest for TEM or SEM analysis are oriented such that the microscope looks down on a {011} "block face." If you orient the specimen's block face so as to attempt to cut {011} cross sections, the diamond knife will touch the Si and immediately propagate a cleave down the nearest {111} plane despite the orientation of the block. As the specimen in the block is constrained by the epoxy holding it, the result is a flake of Si dust from just the surface. When the knife comes around again you get more dust.
Using a microtome to make Si {011}cross sections is like using a screw driver to drive a nail.
Instead of fighting the fact that Si cleaves, why not exploit this fact and prepare really great thin specimens by the microcleaving technique pioneered by McCaffrey and Igor. I'll copy them and invite them to expand on this suggestion. They'll point out that you can make nice thin sections of thin films on Si with a minimum of tools and at room temperature. There is a videotape that shows how to do it.
Ron Anderson
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 } ---------------------------------------------------------------------------- } } I do not know exactly what I can divulge, but they have tried tripod polishing, and ion-milling but I am told they elevate the temperature and the layers need to be prepped and kept at room temps until imaging. } } Does that say enough? I still feel it's a bit irrelevant to the initial question. } } Is it possible to section a Si wafer? Am I correct in being skeptical? Or should I just kill a $4k knife or try an ancient already messed up and questionably anchored diamond in the attempt? } } I just figured someone out there might be able to share a bit of practical experience. } } Thanks! } } Geoff Williams } Leduc Bioimaging Facility Manager } Brown University } } http://www.brown.edu/Facilities/Leduc_Bioimaging_Facility/ } } } } ==============================Original Headers============================== } 8, 29 -- From Geoffrey_Williams-at-brown.edu Tue Aug 22 10:59:55 2006 } 8, 29 -- Received: from draco.services.brown.edu (draco.services.brown.edu [128.148.106.172]) } 8, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MFxtQK026422 } 8, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:59:55 -0500 } 8, 29 -- Received: from ex-gateway1-out.AD.Brown.Edu (ex-gateway1.ad.brown.edu [128.148.21.51]) } 8, 29 -- by draco.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k7MFxs7W006189 } 8, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 11:59:54 -0400 (EDT) } 8, 29 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway1-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); } 8, 29 -- Tue, 22 Aug 2006 11:59:54 -0400 } 8, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 } 8, 29 -- Content-class: urn:content-classes:message } 8, 29 -- MIME-Version: 1.0 } 8, 29 -- Content-Type: text/plain; } 8, 29 -- charset="iso-8859-1" } 8, 29 -- Subject: Sectioning Si, clarification } 8, 29 -- Date: Tue, 22 Aug 2006 11:59:53 -0400 } 8, 29 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F047F6793-at-MAIL1.AD.Brown.Edu} } 8, 29 -- X-MS-Has-Attach: } 8, 29 -- X-MS-TNEF-Correlator: } 8, 29 -- Thread-Topic: Sectioning Si, clarification } 8, 29 -- Thread-Index: AcbGBAF0V914xLqeQPmnA7STgTxe2Q== } 8, 29 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} } 8, 29 -- To: {Microscopy-at-microscopy.com} } 8, 29 -- X-OriginalArrivalTime: 22 Aug 2006 15:59:54.0245 (UTC) FILETIME=[02015B50:01C6C604] } 8, 29 -- X-Brown-Proofpoint: Not Infected } 8, 29 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0608080000 definitions=main-0608220003 } 8, 29 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0608080000 definitions=main-0608220003 } 8, 29 -- Content-Transfer-Encoding: 8bit } 8, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7MFxtQK026422 } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 11, 21 -- From randerson20-at-tampabay.rr.com Tue Aug 22 13:16:47 2006 11, 21 -- Received: from ms-smtp-06.tampabay.rr.com (ms-smtp-06.tampabay.rr.com [65.32.5.136]) 11, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MIGj91015481 11, 21 -- for {Microscopy-at-Microscopy.Com} ; Tue, 22 Aug 2006 13:16:46 -0500 11, 21 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 11, 21 -- by ms-smtp-06.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k7MIGVGD023421; 11, 21 -- Tue, 22 Aug 2006 14:16:37 -0400 (EDT) 11, 21 -- Message-ID: {44EB49FD.2020607-at-tampabay.rr.com} 11, 21 -- Date: Tue, 22 Aug 2006 14:16:29 -0400 11, 21 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 11, 21 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 11, 21 -- MIME-Version: 1.0 11, 21 -- To: Geoffrey_Williams-at-brown.edu, Listserver {Microscopy-at-Microscopy.Com} , 11, 21 -- John McCaffrey {mccaffrey2002-at-rogers.com} , 11, 21 -- Scott Walck {walck-at-southbaytech.com} 11, 21 -- Subject: Re: [Microscopy] Sectioning Si, clarification 11, 21 -- References: {200608221600.k7MG02nF026627-at-ns.microscopy.com} 11, 21 -- In-Reply-To: {200608221600.k7MG02nF026627-at-ns.microscopy.com} 11, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 11, 21 -- Content-Transfer-Encoding: 7bit 11, 21 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
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Email: delsignore-at-gmail.com Name: Anthony Del Signore
Organization: Brown University
Education: Graduate College
Location: Providence, RI, 02865
Title: imaging of cellular aggregates
Question: I am trying to image an aggregate of cells that are producing GFP. I am running into problems when trying to image with Epifluorescent ant Confocal microscopes. I was wondering if I am using the right equipment. It seems that with the epifluor I get alot of deconvolved and out of focus light. With the confocal I am only able to image about 30 microns into the 200 micron spherical aggregate. I get a rim around the spheroid and then black within the aggregate. I am using a 10x objective. Are there any other methods for imaging... I have also heard of flow cytometry and two photon microscopy. Are these valid methods for imaging cellular aggregates emitting GFP? Any help, suggestions would be greatly appreciated. If you need additional information or clarification please let me know as well. Thanks and have a great day.
Metamorph from Molecular Devices works excellent but it is expensive.
You can try ImageJ (free), I have used for less than 50 images.
Zsolt
************************************** Zsolt Lazar Molecular Cytology Core Facility Memorial Sloan-Kettering Cancer Center 415 East 68th Street, ZRC-1838 New York, NY 10021
-----Eredeti üzenet----- Feladó: jds451-at-psu.edu [mailto:jds451-at-psu.edu] Küldve: V 8/20/06 11:04 CÃmzett: Lazar, Zsolt/Sloan-Kettering Institute Másolatot kap: Tárgy: [Microscopy] Software: Montage
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Hi all, I have a friend who needs to make a montage of a large number of optical micrographs. He has about 500 images. Can anybody suggest a PC software package that is easy to use? He is currently using Photoshop. Freeware or something that can be paid for and downloaded would be especially useful. Thanks for any suggestions,
Jennifer Ray Sloppy Materials Science & Engineering, Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 3, 17 -- From jds451-at-psu.edu Sun Aug 20 09:51:57 2006 3, 17 -- Received: from webmail19.cac.psu.edu (webmail19.cac.psu.edu [128.118.1.205]) 3, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7KEpvZI004541 3, 17 -- for {microscopy-at-microscopy.com} ; Sun, 20 Aug 2006 09:51:57 -0500 3, 17 -- Received: (from webmail-at-localhost) 3, 17 -- by webmail19.cac.psu.edu (8.9.3/8.9.0) id KAA20369; 3, 17 -- Sun, 20 Aug 2006 10:51:56 -0400 (EDT) 3, 17 -- Date: Sun, 20 Aug 2006 10:51:56 -0400 (EDT) 3, 17 -- Message-Id: {200608201451.KAA20369-at-webmail19.cac.psu.edu} 3, 17 -- To: microscopy-at-microscopy.com 3, 17 -- Subject: Software: Montage 3, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} 3, 17 -- X-Mailer: Penn State WebMail 3, 17 -- X-Sender: jds451 3, 17 -- X-Originating-IP: 207.255.156.167 3, 17 -- MIME-Version: 1.0 3, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
==============================Original Headers============================== 12, 35 -- From lazarz-at-mskcc.org Tue Aug 22 14:28:47 2006 12, 35 -- Received: from extmail.mskcc.org (mskresolve-9.mskcc.org [140.163.254.134]) 12, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MJSlVI015018 12, 35 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 14:28:47 -0500 12, 35 -- Received: from 140.163.141.235 by extmail.mskcc.org with ESMTP (ESMTP 12, 35 -- Relay); Tue, 22 Aug 2006 15:28:41 -0400 12, 35 -- X-Server-Uuid: 74E28B0A-64D9-4563-A4EB-AECFC40E1CE3 12, 35 -- Received: from SMSKPEXMBX6.MSKCC.ROOT.MSKCC.ORG ([140.163.141.233]) by 12, 35 -- smskpexsmtp2.MSKCC.ROOT.MSKCC.ORG with Microsoft 12, 35 -- SMTPSVC(5.0.2195.6713); Tue, 22 Aug 2006 15:28:41 -0400 12, 35 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 12, 35 -- content-class: urn:content-classes:message 12, 35 -- MIME-Version: 1.0 12, 35 -- Subject: RE: [Microscopy] Software: Montage 12, 35 -- Date: Tue, 22 Aug 2006 15:28:41 -0400 12, 35 -- Message-ID: {0534480A8E1A0F4187DC6FFC3DDEECFF105D08-at-smskpexmbx6.mskcc.root.mskcc.org} 12, 35 -- X-MS-Has-Attach: 12, 35 -- X-MS-TNEF-Correlator: 12, 35 -- Thread-Topic: [Microscopy] Software: Montage 12, 35 -- Thread-Index: AcbEafXouMo6JtQqTTaLoK6w3FUoHwBtrb+m 12, 35 -- References: {200608201504.k7KF4Sea013592-at-ns.microscopy.com} 12, 35 -- From: LazarZ-at-mskcc.org 12, 35 -- To: microscopy-at-microscopy.com 12, 35 -- X-OriginalArrivalTime: 22 Aug 2006 19:28:41.0498 (UTC) 12, 35 -- FILETIME=[2CD477A0:01C6C621] 12, 35 -- X-TMWD-Spam-Summary: TS=20060822192842; SEV=2.0.1; DFV=A2006082207; 12, 35 -- IFV=2.0.4,4.0-8; RPD=4.00.0004; ENG=IBF; 12, 35 -- RPDID=303030312E30413031303230342E34344542353944432E303034392D412D; 12, 35 -- CAT=NONE; CON=NONE 12, 35 -- X-MMS-Spam-Filter-ID: A2006082207_4.00.0004_4.0-8 12, 35 -- X-WSS-ID: 68F585630YC1526875-01-01 12, 35 -- Content-Type: text/plain; 12, 35 -- charset=utf-8 12, 35 -- Content-Transfer-Encoding: 8bit 12, 35 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k7MJSlVI015018 ==============================End of - Headers==============================
Listers, I had a response back on my Si microtoming tome from a biologist who wrote: "People like me (and Geoff) read this and go "OooKay ... what's a {111} plane and how would I know how I have such a thing aligned for anything, much less microtomy?"
Answer: Any Si semiconductor wafer you find will most likely have a {001} direction perpendicular to its surface. If it is a whole wafer there is probably a flat and a notch ground into the wafers edge at right angles to each other. The families of crystal {011} planes are oriented at 45 and 90 degrees with respect to each other. The flat is parallel to one of the {011} planes in the family. The notch is perpendicular to a second {011} plane at 90 degrees wrt to the flat. If it is a processed Si chip, look at the x, y coordinates defined on the surface of the chip. They are parallel to {011} planes. {111} planes are oriented at 35 and 90 degrees with respect to {011} planes. Try cleaving the wafer at 45 or 90 degrees wrt to the flat or notch. When you get a piece with a 35 degree slope wrt to the surface, the slope part will be a {111} face. OR, just hit the Si wafer with a hammer and look for a piece with shiny 35 degree sloped edges wrt the surface.
BUT, my biological friends, why are you making your heads hurt by reading this? The point of my original post was to explain why you won't want to fracture the edge of your diamond knife creating Si dust. Now you know that the dust particles will have 35 degree sloped sides! You can't microtome Si. Geoff, go read McCaffrey's paper on microcleaving. When Scott weighs in, he'll provide the reference, I hope.
Ron Anderson
==============================Original Headers============================== 5, 17 -- From randerson20-at-tampabay.rr.com Tue Aug 22 14:49:20 2006 5, 17 -- Received: from ms-smtp-01.tampabay.rr.com (ms-smtp-01.tampabay.rr.com [65.32.5.131]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MJnKuA025403 5, 17 -- for {Microscopy-at-Microscopy.Com} ; Tue, 22 Aug 2006 14:49:20 -0500 5, 17 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 5, 17 -- by ms-smtp-01.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k7MJnGaX023286 5, 17 -- for {Microscopy-at-Microscopy.Com} ; Tue, 22 Aug 2006 15:49:19 -0400 (EDT) 5, 17 -- Message-ID: {44EB5FBA.2050008-at-tampabay.rr.com} 5, 17 -- Date: Tue, 22 Aug 2006 15:49:14 -0400 5, 17 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 5, 17 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 5, 17 -- MIME-Version: 1.0 5, 17 -- To: Listserver {Microscopy-at-Microscopy.Com} 5, 17 -- Subject: Microtoming Silicon 5, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 17 -- Content-Transfer-Encoding: 7bit 5, 17 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
I've never tried it on embedded and sectioned tissue, but wouldn't iodine work? It works well enough in fresh tissue, staining starch grains purple in light microscopy.
I do know that we've had a lot of trouble imaging tissue with starch granules in it, as they refract light, and mess up the signal in our confocal and tomography images. So polarized light might also work very well. Again I've never tried it.
I'm not sure if this helps, but there you go. Robin
--------------------------------- Robin Young, M.Sc. PhD Candidate Samuels and Haughn Labs Dept of Botany, UBC 6270 University Blvd. Vancouver, BC V6T 1Z4 fax: 604-822-6089
==============================Original Headers============================== 4, 27 -- From youngre-at-interchange.ubc.ca Tue Aug 22 15:04:55 2006 4, 27 -- Received: from mta4.mail-relay.ubc.ca (mta4.mail-relay.ubc.ca [137.82.45.8]) 4, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MK4s78003559 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 15:04:55 -0500 4, 27 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 4, 27 -- by mta4.mail-relay.ubc.ca (8.12.11.20060308/8.12.11) with ESMTP id k7MK4icq006821 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 13:04:44 -0700 (PDT) 4, 27 -- (envelope-from youngre-at-interchange.ubc.ca) 4, 27 -- Received: from [137.82.136.201] (robin.botany.ubc.ca [137.82.136.201]) 4, 27 -- by smtp.interchange.ubc.ca 4, 27 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 4, 27 -- with ESMTPSA id {0J4F00FD213W8U-at-smtp.interchange.ubc.ca} for 4, 27 -- microscopy-at-microscopy.com; Tue, 22 Aug 2006 13:04:44 -0700 (PDT) 4, 27 -- Date: Tue, 22 Aug 2006 13:04:44 -0700 4, 27 -- From: Robin Young {youngre-at-interchange.ubc.ca} 4, 27 -- Subject: Re: staining starch in sections 4, 27 -- To: microscopy-at-microscopy.com 4, 27 -- Message-id: {2EDB6CAF-30C7-4B9C-B0FB-1062C1634EB9-at-interchange.ubc.ca} 4, 27 -- MIME-version: 1.0 (Apple Message framework v752.2) 4, 27 -- X-Mailer: Apple Mail (2.752.2) 4, 27 -- Content-type: text/plain; format=flowed; delsp=yes; charset=US-ASCII 4, 27 -- Content-transfer-encoding: 7bit 4, 27 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.8.22.115442 4, 27 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 4, 27 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P1_5 0, __C230066_P5 0, __CP_MEDIA_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 4, 27 -- X-Spam-Level: 4, 27 -- X-Spam-Flag: No ==============================End of - Headers==============================
Looking to digitize analog NTSC video tape (Hi-8 and VHS, so composite video, not S-video or component). Would like to stream the analog signal right in to digital format for later image analysis (so only need the video data no sound), and would like to work whole 60min tapes at a time. Since we’re looking to collect measurement data from video still images the higher the quality preserved the better. So looking for *specific* recommend ations as to the best method for doing this (specific as in model numbers).
Computer system (Windows PC) we have for this: has 1394, USB, 16x PCIe, x4 PCIe, 64-bit PCI, and 32-Bit PCI, with a RAID Array. Adobe Premiere reports system can sustain over 35MB/sec throughput.
Possible solutions:
1) A/D capture card, o.k., which one for running under Windows XP? Capturing composite NTSC video seems to be “old-tech” these days. Seemingly easy to find HDTV capture cards but not NTSC.
2) A/D capture “widgets” = Composite video in and IEEE 1394 output or USB? Seems like a very cost effective solution, can anyone recommend a model?
3) Using either a Digital8 camcorder or something like a Sony Digital8 Video Walkman to play the “tapes” out to 1394 and input the 1394 to the computer. (Note: Digital8 Players will play Hi-8 tapes). Will this work?
Video / Image Analysis: Once digitized the “video” will be broken up into digital stills (yes, 18 hours of still images at 29 frames / sec.). We actually only need 1 image from approximately every 4 seconds of video. Every 4 seconds the video image changes to one of 16 “views”. We will then sort the images into the individual 16 “views”. Each of the 16 “views” shows 7 developing seedlings. We will then analyze each of the 16 “views” (or sets of seedlings) for growth changes through time. Oh, yes after each set of 16 “views” the tape is “paused” for 10 mins, and then repeats the 16 view capture sequence again. So basically it is time lapse imaging, of 16 different subjects, all interleaved on one video stream.
And why collect data this way you ask? Because the data collection is all done automatically and remotely . . . From a minimum of 355 miles up in orbit on the International Space Station, on an automated growth and imaging system, which does not have room for 16 video cameras and video capture systems.
Any suggestions would be greatly appreciated.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 16, 24 -- From edelmare-at-muohio.edu Tue Aug 22 15:12:24 2006 16, 24 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MKCNVa013909 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 15:12:24 -0500 16, 24 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 16, 24 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7MKCBHt014555 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 16:12:12 -0400 16, 24 -- Received: from emf03 ([134.53.14.97]) 16, 24 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7MKC9EJ014949 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 16:12:09 -0400 16, 24 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 16, 24 -- To: microscopy-at-Microscopy.com 16, 24 -- Date: Tue, 22 Aug 2006 16:14:11 -0400 16, 24 -- MIME-Version: 1.0 16, 24 -- Content-type: text/plain; charset=ISO-8859-1 16, 24 -- Subject: Looking for some video capture recommendations 16, 24 -- Message-ID: {44EB2D53.3411.1471D88D-at-localhost} 16, 24 -- Priority: normal 16, 24 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 16, 24 -- X-Real-ConnectIP: 134.53.14.97 16, 24 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.67 16, 24 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 16, 24 -- Content-Transfer-Encoding: 8bit 16, 24 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k7MKCNVa013909 ==============================End of - Headers==============================
I've used Iodine-Potassium Iodide solution (dissolve 2g of potassium iodide in 100 ml water then dissolve 0.2g of iodine in this).
Ian
Ian Hallett Microscopy HortResearch Mt Albert Research Centre Private Bag 92 169, Mt Albert Auckland, New Zealand +64-9-815 4200 ext 7002
-----Original Message----- X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu] Sent: Wednesday, 23 August 2006 2:26 a.m. To: Ian Hallett
Greetings, Can anyone tell me a reliable method to stain starch grains in sections of plant tissue embedded in Spurr's resin? Also useful to know whether polarized light would be suitable. I hope this is easy and I get lots of replies!
As ever, Tobias -- _ ____ __ ____ / \ / / \ / \ \ Tobias I. Baskin / / / / \ \ \ Biology Department /_ / __ /__ \ \ \__ 611 N. Pleasant St. / / / \ \ \ University of Massachusetts / / / \ \ \ Amherst, MA, 01003 / / ___ / \ \__/ \ ____ http://www.bio.umass.edu/biology/baskin/ Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243
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==============================Original Headers============================== 15, 30 -- From IHallett-at-hortresearch.co.nz Tue Aug 22 15:58:34 2006 15, 30 -- Received: from hortresearch.co.nz (mscan.hortresearch.co.nz [202.36.134.15]) 15, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MKwXec024715 15, 30 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 15:58:34 -0500 15, 30 -- Received: from aklexf01.hort.net.nz ([10.16.1.14]) by hortresearch.co.nz 15, 30 -- with HortResearch; Wed, 23 Aug 2006 09:09:55 +1200 15, 30 -- Received: from AKLEXB01.hort.net.nz ([10.16.1.15]) by aklexf01.hort.net.nz 15, 30 -- with Microsoft SMTPSVC(6.0.3790.1830); Wed, 23 Aug 2006 08:58:00 +1200 15, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 15, 30 -- Content-class: urn:content-classes:message 15, 30 -- MIME-Version: 1.0 15, 30 -- Content-Type: text/plain; 15, 30 -- charset=us-ascii 15, 30 -- Subject: RE: [Microscopy] staining starch in sections 15, 30 -- Date: Wed, 23 Aug 2006 08:57:59 +1200 15, 30 -- Message-ID: {D3BAD63C088F3C48AEB385E881359F870214C10A-at-AKLEXB01.hort.net.nz} 15, 30 -- In-Reply-To: {200608221425.k7MEPxFe020693-at-ns.microscopy.com} 15, 30 -- X-MS-Has-Attach: 15, 30 -- X-MS-TNEF-Correlator: 15, 30 -- Thread-Topic: [Microscopy] staining starch in sections 15, 30 -- Thread-Index: AcbF9uX7+gwr9oxdQyCAVrJmBhwvjQANh36A 15, 30 -- From: "Ian Hallett" {IHallett-at-hortresearch.co.nz} 15, 30 -- To: {microscopy-at-microscopy.com} 15, 30 -- X-OriginalArrivalTime: 22 Aug 2006 20:58:00.0040 (UTC) 15, 30 -- FILETIME=[A6C51680:01C6C62D] 15, 30 -- X-imss-version: 2.042 15, 30 -- X-imss-result: Passed 15, 30 -- X-imss-approveListMatch: *-at-hortresearch.co.nz 15, 30 -- Content-Transfer-Encoding: 8bit 15, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7MKwXec024715 ==============================End of - Headers==============================
Have you considered the Thièry method, using periodate oxidation of vicinal hydroxyls in carbohydrates, reacting resulting aldehydes with thiocarbohydrazide or thiosemicarbazide followed by silver proteinate? It may not be the easiest method, though, but it works really well.
I don't have the original reference (1964 I believe), but it is a.o. described in JHC Volume 33, Issue 10, pp. 1007-1014, 1985
Jan Leunissen
Research Advisor Electron Microcopy Dept Anatomy / Structural Biology http://anatomy.otago.ac.nz/
==============================Original Headers============================== 9, 18 -- From leunissen-at-aurion.nl Tue Aug 22 16:05:30 2006 9, 18 -- Received: from fep04.xtra.co.nz (fep04.xtra.co.nz [210.54.141.242]) 9, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7ML5TnC002729 9, 18 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 16:05:30 -0500 9, 18 -- Received: from [192.168.1.50] (really [222.152.176.211]) 9, 18 -- by fep04.xtra.co.nz with ESMTP 9, 18 -- id {20060822210523.KTBB6771.fep04.xtra.co.nz-at-[192.168.1.50]} 9, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 23 Aug 2006 09:05:23 +1200 9, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) 9, 18 -- Message-Id: {C1FD8ECD-C854-4933-AA03-0FE421FB6BC6-at-aurion.nl} 9, 18 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 9, 18 -- To: Microscopy-at-microscopy.com 9, 18 -- From: Jan Leunissen {leunissen-at-aurion.nl} 9, 18 -- Subject: Re: [Microscopy] staining starch in sections 9, 18 -- Date: Wed, 23 Aug 2006 09:05:22 +1200 9, 18 -- X-Mailer: Apple Mail (2.752.2) 9, 18 -- Content-Transfer-Encoding: 8bit 9, 18 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7ML5TnC002729 ==============================End of - Headers==============================
Iodine would surely stain starch molecules. Many years ago, I used iodine to stain blends of polypropylene and ethylene vinyl alcohol (EVOH) for analysis by SEM and TEM. The iodine did provide excellent initial selective contrast for the EVOH. Unfortunately, iodine is volatile in the vacuum of the microscope. The iodine bound by the EVOH dissipated sufficiently rapidly that I saw a significant reduction in contrast during the microscopy session.
A better stain for examination of starch in electron microscopes probably osmium tetroxide. Osmium binds the hydroxyl sites irreversibly, thus providing excellent heavy metal contrast in the preparation.
Good luck,
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
youngre-at-interc hange.ubc.ca To gary.m.brown-at-exxonmobil.com 08/22/06 03:07 cc PM Subject [Microscopy] Re: staining starch in Please respond sections to youngre-at-interc hange.ubc.ca
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I've never tried it on embedded and sectioned tissue, but wouldn't iodine work? It works well enough in fresh tissue, staining starch grains purple in light microscopy.
I do know that we've had a lot of trouble imaging tissue with starch granules in it, as they refract light, and mess up the signal in our confocal and tomography images. So polarized light might also work very well. Again I've never tried it.
I'm not sure if this helps, but there you go. Robin
--------------------------------- Robin Young, M.Sc. PhD Candidate Samuels and Haughn Labs Dept of Botany, UBC 6270 University Blvd. Vancouver, BC V6T 1Z4 fax: 604-822-6089
==============================Original Headers============================== 4, 27 -- From youngre-at-interchange.ubc.ca Tue Aug 22 15:04:55 2006 4, 27 -- Received: from mta4.mail-relay.ubc.ca (mta4.mail-relay.ubc.ca [137.82.45.8]) 4, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MK4s78003559 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 15:04:55 -0500 4, 27 -- Received: from mta2.interchange.ubc.ca (mta2.interchange.ubc.ca [142.103.145.70]) 4, 27 -- by mta4.mail-relay.ubc.ca (8.12.11.20060308/8.12.11) with ESMTP id k7MK4icq006821 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 13:04:44 -0700 (PDT) 4, 27 -- (envelope-from youngre-at-interchange.ubc.ca) 4, 27 -- Received: from [137.82.136.201] (robin.botany.ubc.ca [137.82.136.201]) 4, 27 -- by smtp.interchange.ubc.ca 4, 27 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 4, 27 -- with ESMTPSA id {0J4F00FD213W8U-at-smtp.interchange.ubc.ca} for 4, 27 -- microscopy-at-microscopy.com; Tue, 22 Aug 2006 13:04:44 -0700 (PDT) 4, 27 -- Date: Tue, 22 Aug 2006 13:04:44 -0700 4, 27 -- From: Robin Young {youngre-at-interchange.ubc.ca} 4, 27 -- Subject: Re: staining starch in sections 4, 27 -- To: microscopy-at-microscopy.com 4, 27 -- Message-id: {2EDB6CAF-30C7-4B9C-B0FB-1062C1634EB9-at-interchange.ubc.ca} 4, 27 -- MIME-version: 1.0 (Apple Message framework v752.2) 4, 27 -- X-Mailer: Apple Mail (2.752.2) 4, 27 -- Content-type: text/plain; format=flowed; delsp=yes; charset=US-ASCII 4, 27 -- Content-transfer-encoding: 7bit 4, 27 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.8.22.115442 4, 27 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 4, 27 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P1_5 0, __C230066_P5 0, __CP_MEDIA_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 4, 27 -- X-Spam-Level: 4, 27 -- X-Spam-Flag: No ==============================End of - Headers==============================
==============================Original Headers============================== 23, 20 -- From gary.m.brown-at-exxonmobil.com Tue Aug 22 16:08:45 2006 23, 20 -- Received: from hoespc02.exxonmobil.com (hoespc02.exxonmobil.com [192.67.48.39]) 23, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7ML8iET011132 23, 20 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 16:08:45 -0500 23, 20 -- Received: from hounmg03.NA.XOM.COM (hounmg03.na.xom.com [158.35.101.41]) 23, 20 -- by hoespc02.exxonmobil.com (Switch-3.1.7/Switch-3.1.7) with ESMTP id k7ML8dQP027536; 23, 20 -- Tue, 22 Aug 2006 16:08:39 -0500 (CDT) 23, 20 -- In-Reply-To: {200608222007.k7MK7Ct3007202-at-ns.microscopy.com} 23, 20 -- Subject: Re: [Microscopy] Re: staining starch in sections 23, 20 -- Importance: 23, 20 -- To: youngre-at-interchange.ubc.ca, microscopy-at-microscopy.com 23, 20 -- Cc: donald.a.winesett-at-exxonmobil.com 23, 20 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 23, 20 -- Message-ID: {OF254CE6BC.DC284982-ON862571D2.0072DBD2-862571D2.0074255C-at-exxonmobil.com} 23, 20 -- From: gary.m.brown-at-exxonmobil.com 23, 20 -- Date: Tue, 22 Aug 2006 16:08:37 -0500 23, 20 -- X-MIMETrack: Serialize by Router on Hounmg03.na.xom.com/S/ExxonMobil(652FP1HF193|March 23, 20 -- 02, 2006) at 08/22/2006 04:08:50 PM 23, 20 -- MIME-Version: 1.0 23, 20 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Thank you, Ron and Henk, for the plug. Sorry, I was on vacation or else I would have answered this.
I'd like to address the MicroCleave(TM) preparation at the end of this discussion so that it is closest to the disclaimer that I am going to put on it.
SDW on Microtoming: I've seen some images that Tom Malis collected of microtomed Si samples from different sources. True, you get a lot of mostly dust as Ron says, but apparently when you hit it using the right conditions, you can get little cracked pieces that compose the ribbon that have the film intact on them. If you are interested in only the film, you have a chance of getting what you need. However, for the most part, there are other methods for making Si cross sections that are much easier and much more likely for producing a usable sample.
SDW on Ion Milling: I have worked with thin Au-Ge-Ni multi-layers on GaAs in the past and you do have to worry about heating the sample as you indicate. You can use superglue in place of wax; you just have to be patient to get rid of the glue. If you ion mill your sample, you really have to worry about heating the sample during ion milling. The answer there is to use low angle, low energy, and it wouldn't hurt to cool the sample with a liquid nitrogen cooled stage. Since it is a competitor's product, I will not mention the name, but there is one popular ion mill on the market that you have to be particularly careful with. This ion mill does not have a sample holder that has a large heat sink and has a relatively poor thermal path through the holder. If you have this type of ion mill, you must significantly de-tune the ion guns or you can heat your samples very high even at low milling angles. Unfortunately, this "de-tuning" by lowering the ion energy and limiting the ion beam current will significantly impact the ion milling rate. I have melted glass cross sections with this ion mill. I have used other ion mills with the glass and for the most part, they are better in terms of not heating the glass samples simply because the stages dissipate the heat better, but you still have to be careful. In all of the conventional ion mills, i.e. non low-energy types, the beam current drops to essentially zero below about 3 kV. There are a number of studies that have been done concerning the heating of samples in ion mills. You can contact me off-line to talk about the low energy guns in the ion mills that we sell.
SDW on MicroCleaving: OK, let me sound off for the MicroCleave(TM), also known as the Small Angle Cleavage Technique. (You can even consider this coming from Igor Slobinsky and Dr. Cleavinstein so maybe I won't get in trouble for being too commercial.) I thought that this technique is the neatest thing since sliced bread (or sliced samples) when I started using it and I still do. This technique produces the best cross section samples of any available technique because there is simply no ion damage produced. In fact, a major disadvantage of these types of samples is that if the samples are fully crystalline and you are using a field emission gun equipped microscope, then there is no amorphous regions to help the microscopist focus and stigmate with. I suspect that with the new C(s)-corrected machines, more and more people will be discovering this technique.
For the metallization samples, the MicroCleave(TM) technique will work as long as the layers are very thin. Generally, they should be less than 1000A. They also must be fully adherent to the substrate and to the other layers. For as-deposited Au-Ge-Ni on GaAs samples with e-beam evaporated Ge layers that I mentioned above, I found that the Ge layers were weak and would split within the layer during the cleave. I've never looked at Ge layers put down by other methods, so I don't know whether that is a universal trait of Ge layers or not. If we heat treated them to alloy the layers, I had no such problem. Even a slight anneal where there was no apparent mixing of the layer materials would help prevent the Ge layer from splitting. I suspect that we might have been getting a little densification with the slight anneal. What happens with these samples during the procedure is that the thinnest parts of the samples would consist of any layers below the Ge and about half the Ge layer. Further back, all of the layers would be intact, but thicker. I can send a copy of a presentation on the MicroCleave technique that John McCaffrey and I did in "Cleave-Land" at an M&M meeting in 1997 to anyone that wants one. There are two places in the presentation that addresses temperature sensitive samples. First, as you point out, use superglue and then wait for the superglue to dissolve when finished instead of using the low temperature melting wax. The other place is that for the mounting epoxy, you need a viscous and slow setting epoxy to mount the samples onto the specially bent Veco 2x1 tabbed grids. Normally, Epoxy Technology's H-22 silver filled epoxy is used, but that requires heat to set. The 90 minute Super strength type of two part epoxies work well, but you have to give them a full day to set up. They are also nonconductive, but that didn't seem to be much of a problem. I've tried a number of different brands, and they seemed to work, but they have to be the 90 minute working time ones. Other epoxies flowed too much and did not stay in position to hold the sample. I also experimented with fillers for the epoxy with no success at all. There is also a temperature spike when an epoxy cures. When I used this on temperature sensitive samples, I tried to minimize the amount of epoxy on the grids so that the grid could act as a heat sink for the exothermic reaction and the samples would experience the minimum of a thermal excursion.
One other point of caution for about Si samples prepared this way. The samples can become very very thin and basically disappear into nothingness at the edges. I had a pulsed laser deposited diamond like carbon film that was deposited onto silicon and prepared this way that was too thin at the edges to do electron energy loss spectroscopy on and get sufficient signal! That's thin! What it means is that the samples prepared this way can be heated by a focused beam because there isn't enough thermal path to extract heat from the exposed area.
Here is a list of the references that I have. The "Update" paper by John and me also has an accompanying presentation that I can send to people in a PDF format that I can send to anyone that is interested. This presentation and the MRS paper by us have a pictorial outline of the technique in it. There are a lot of hands-on tips and tricks and details that we tried to put into the paper so that someone could try the technique without reinventing the wheel.
1) "Small Angle Cleavage of Semiconductors for TEM", J.P. McCaffrey, Ultramicroscopy 38, 1991, pp:149-157 2) "TEM Samples of Semiconductors Prepared by a Small Angle Cleavage Technique", J.P. McCaffrey, Mat. Res. Soc. Symp. Proc., Vol.254, 1992, pp:109-120 3) John P. McCaffrey, Microscopy Research and Technique, 24, (1993) 180. 4) "A Simplified Method for Modifying TEM Copper Grids For Use with the Small Angle Cleavage Technique", Scott D. Walck, Microscopy Today, (96-4), (1996). 5) The Small Angle Cleavage Technique Applied to Coatings and Thin Films, S. D. Walck and J. P. McCaffrey, Thin Solid Films, 308-309, (1997) 399. 6) "The Small Angle Cleavage Technique: An Update", Scott D. Walck, John P. McCaffrey, Mat.Res.Soc.Symp.Proc., Vol.480,1997,pp:149-170
OK, here is the disclaimer that I hope will keep me out of trouble. South Bay Technology Inc. makes and sells the Microcleave(TM) Kit. SBT also sells the Technoorg-Linda line of ion mills with the low energy ion gun.
-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: randerson20-at-tampabay.rr.com [mailto:randerson20-at-tampabay.rr.com] Sent: Tuesday, August 22, 2006 11:20 AM To: Walck-at-SouthBayTech.com
Geoff,
No you can't "cut" Si with a diamond knife. Si cleaves very easily. When the knife touches the Si, the Si cleaves along one of its preferred {111} crystallographic cleaving planes. That might be fine if you orient the Si such that a Si {111} crystallographic direction is perpendicular to the block face. Then if you are VERY lucky, you MIGHT get a cleaved piece of Si oriented with {111} faces (at the risk of your knife). Any article that I've seen that claims to have microtomed Si has shown lots of pictures of tiny {111} Si sections.
Assuming that you are successful, so what! There is no, or very little interest, in anything in the semiconductor world or in potentially oriented thin films deposited on Si, that is best displayed on top of a Si cross section with a {111} block face orientation. The angles are wrong.
Si wafers in semiconductor processing are almost all with (or very close to) a Si {001} direction perpendicular to the Si wafer surface. Devices fabricated in, or patterns deposited on, these wafers are not randomly oriented. They are almost always aligned with {011} planes perpendicular to the Si surface, i.e. the x, y orientations of the devices and patterns are parallel to {011} directions in the Si surface. Cross sections of interest for TEM or SEM analysis are oriented such that the microscope looks down on a {011} "block face." If you orient the specimen's block face so as to attempt to cut {011} cross sections, the diamond knife will touch the Si and immediately propagate a cleave down the nearest {111} plane despite the orientation of the block. As the specimen in the block is constrained by the epoxy holding it, the result is a flake of Si dust from just the surface. When the knife comes around again you get more dust.
Using a microtome to make Si {011}cross sections is like using a screw driver to drive a nail.
Instead of fighting the fact that Si cleaves, why not exploit this fact and prepare really great thin specimens by the microcleaving technique pioneered by McCaffrey and Igor. I'll copy them and invite them to expand on this suggestion. They'll point out that you can make nice thin sections of thin films on Si with a minimum of tools and at room temperature. There is a videotape that shows how to do it.
Ron Anderson
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 } ---------------------------------------------------------------------- } ------ } } I do not know exactly what I can divulge, but they have tried tripod polishing, and ion-milling but I am told they elevate the temperature and the layers need to be prepped and kept at room temps until imaging. } } Does that say enough? I still feel it's a bit irrelevant to the initial question. } } Is it possible to section a Si wafer? Am I correct in being skeptical? Or should I just kill a $4k knife or try an ancient already messed up and questionably anchored diamond in the attempt? } } I just figured someone out there might be able to share a bit of practical experience. } } Thanks! } } Geoff Williams } Leduc Bioimaging Facility Manager } Brown University } } http://www.brown.edu/Facilities/Leduc_Bioimaging_Facility/ } } } } ==============================Original } Headers============================== } 8, 29 -- From Geoffrey_Williams-at-brown.edu Tue Aug 22 10:59:55 2006 8, } 29 -- Received: from draco.services.brown.edu (draco.services.brown.edu [128.148.106.172]) } 8, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MFxtQK026422 } 8, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:59:55 -0500 } 8, 29 -- Received: from ex-gateway1-out.AD.Brown.Edu (ex-gateway1.ad.brown.edu [128.148.21.51]) } 8, 29 -- by draco.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k7MFxs7W006189 } 8, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 11:59:54 -0400 (EDT) } 8, 29 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway1-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); } 8, 29 -- Tue, 22 Aug 2006 11:59:54 -0400 } 8, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 29 } -- Content-class: urn:content-classes:message 8, 29 -- MIME-Version: } 1.0 8, 29 -- Content-Type: text/plain; } 8, 29 -- charset="iso-8859-1" } 8, 29 -- Subject: Sectioning Si, clarification 8, 29 -- Date: Tue, 22 } Aug 2006 11:59:53 -0400 8, 29 -- Message-ID: } {A1A84D541C161C4C988B4E0FB38F5A0F047F6793-at-MAIL1.AD.Brown.Edu} } 8, 29 -- X-MS-Has-Attach: } 8, 29 -- X-MS-TNEF-Correlator: } 8, 29 -- Thread-Topic: Sectioning Si, clarification 8, 29 -- } Thread-Index: AcbGBAF0V914xLqeQPmnA7STgTxe2Q== 8, 29 -- From: } "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 8, 29 -- To: } {Microscopy-at-microscopy.com} 8, 29 -- X-OriginalArrivalTime: 22 Aug } 2006 15:59:54.0245 (UTC) FILETIME=[02015B50:01C6C604] 8, 29 -- } X-Brown-Proofpoint: Not Infected 8, 29 -- X-Proofpoint-Spam-Details: } rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 } reason=safe engine=3.1.0-0608080000 definitions=main-0608220003 8, 29 } -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam } policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe } engine=3.1.0-0608080000 definitions=main-0608220003 8, 29 -- } Content-Transfer-Encoding: 8bit 8, 29 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id k7MFxtQK026422 } ==============================End of - } Headers============================== } } }
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==============================Original Headers============================== 29, 22 -- From walck-at-southbaytech.com Tue Aug 22 18:18:57 2006 29, 22 -- Received: from ylpvm43.prodigy.net (ylpvm43-ext.prodigy.net [207.115.57.74]) 29, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MNIv4K025370 29, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 18:18:57 -0500 29, 22 -- X-ORBL: [64.169.193.90] 29, 22 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 29, 22 -- by ylpvm43.prodigy.net (8.13.7 out spool5000 dk/8.13.7) with ESMTP id k7MNIsbi030444; 29, 22 -- Tue, 22 Aug 2006 19:18:55 -0400 29, 22 -- From: "Scott Walck" {walck-at-southbaytech.com} 29, 22 -- To: {Microscopy-at-microscopy.com} 29, 22 -- Cc: {randerson20-at-tampabay.rr.com} , "Henk Colijn" {colijn.1-at-osu.edu} 29, 22 -- Subject: RE: [Microscopy] Re: Sectioning Si, clarification 29, 22 -- Date: Tue, 22 Aug 2006 16:19:36 -0700 29, 22 -- Message-ID: {00a301c6c641$6f4255b0$7801a8c0-at-dynamicbl8uno3} 29, 22 -- MIME-Version: 1.0 29, 22 -- Content-Type: text/plain; 29, 22 -- charset="us-ascii" 29, 22 -- Content-Transfer-Encoding: 7bit 29, 22 -- X-Mailer: Microsoft Office Outlook 11 29, 22 -- In-Reply-To: {200608221820.k7MIKOHm021597-at-ns.microscopy.com} 29, 22 -- Thread-Index: AcbGF6KHgJf2sp57RhCWrZ22d+K1vQACaacw 29, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
If you don't want 16-bit TIFF, why not just save it as 8-bits? SaveAs/Options/Save16-bitAs8-bit.
Save the 16-bit as the original and save the 8-bit from analySIS as the 8-bit version. Dynamic range will be reduced but pixel loss will be reduced.
gary g.
At 08:24 AM 8/22/2006, you wrote:
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Not being a TEM person, I'm exactly sure what you need to have done.
Si wafers are easy to cut using a carbide tip pens available from Home Depot (~$6). You can buy pre-cut wafers from Ted Pella but you will get a lot of little die--probably more than you need.
The easiest way to cut coated Si material is to cleave it. This is perfect for cross section analysis.
If you don't get the Si material's lattice lined up properly, trying any variable cutting method will yield a pile of Si dust. You must be parallel or perpendicular to the wafer flat.
gary g.
At 08:33 AM 8/22/2006, you wrote:
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==============================Original Headers============================== 12, 22 -- From gary-at-gaugler.com Tue Aug 22 18:58:22 2006 12, 22 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7MNwLpA013869 12, 22 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 18:58:22 -0500 12, 22 -- Received: (qmail 26517 invoked from network); 22 Aug 2006 16:58:20 -0700 12, 22 -- Received: by simscan 1.1.0 ppid: 26512, pid: 26515, t: 0.2015s 12, 22 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 22 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 22 -- by qsmtp1 with SMTP; 22 Aug 2006 16:58:20 -0700 12, 22 -- Message-Id: {7.0.1.0.2.20060822165222.0253aa78-at-gaugler.com} 12, 22 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 22 -- Date: Tue, 22 Aug 2006 16:58:21 -0700 12, 22 -- To: Geoffrey_Williams-at-brown.edu 12, 22 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 22 -- Subject: Re: [Microscopy] Sectioning Si Wafers 12, 22 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 22 -- In-Reply-To: {200608221533.k7MFXQG0008602-at-ns.microscopy.com} 12, 22 -- References: {200608221533.k7MFXQG0008602-at-ns.microscopy.com} 12, 22 -- Mime-Version: 1.0 12, 22 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 12, 22 -- Content-Transfer-Encoding: 8bit 12, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7MNwLpA013869 ==============================End of - Headers==============================
I have a montage assembler that is based on Windows Explorer. The individual images must be of consistent size, have overlap between adjacent images and not exceed total available memory.
If you wish to see a sample, can you send me some files? 10x10 (100) images ought to show if this program will work.
The data so far has been collected on a Prior automated stage.
There is no user manual and the software could be free. But it should be tested to be sure that it will work for your application.
gary g.
At 07:55 AM 8/20/2006, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Tue Aug 22 19:04:39 2006 11, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7N04clj024143 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 19:04:39 -0500 11, 20 -- Received: (qmail 29961 invoked from network); 22 Aug 2006 17:04:37 -0700 11, 20 -- Received: by simscan 1.1.0 ppid: 29958, pid: 29959, t: 0.1654s 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; 22 Aug 2006 17:04:37 -0700 11, 20 -- Message-Id: {7.0.1.0.2.20060822165931.02559ba8-at-gaugler.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 20 -- Date: Tue, 22 Aug 2006 17:04:38 -0700 11, 20 -- To: jds451-at-psu.edu 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] Software: Montage 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200608201455.k7KEtlmP007749-at-ns.microscopy.com} 11, 20 -- References: {200608201455.k7KEtlmP007749-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
On Aug 22, 2006, at 11:46, voyles-at-engr.wisc.edu wrote:
} There is also a potentially serious drawback to the Photoshop } procedure } you suggest. Photoshop discards the original grey-scale values of the } image when you use "Auto Levels", which breaks the quantitative } connection between the grey-scale value in the image and the absolute } number of electrons that struck the detector. This will often make } quantitative analysis of the electron image or diffraction pattern } impossible and makes it difficult to even judge the relative average } brightness of two different images to guess, for example, the sample } thickness.
This is true. However, it is trivial to rectify the problem that he's having, which is that his system is saving 12-bit TIFFs. Simply use the Convolve function in Photoshop to multiply the image by 16 uniformly. This preserves the quantitative connection while allowing you to visually observe the image. (One notes that there was a very simple tutorial for this in Microscopy Today about two years ago... *g*)
} Software developed for photographs and desktop } publishing is not generally the best choice for dealing with } scientific } data.
I disagree with this statement. You can be perfectly quantitative in Photoshop, and in fact many people are (see all the Fovea Pro users, for example.) The issue at hand is one of needing to understand what one is doing to an image. But that issue is true no matter what software you're using. You can screw up data in ImagePro just as easily as you can in Photoshop.
Brent
-- Brent Neal Research Physicist Milliken Research Corporation
==============================Original Headers============================== 16, 27 -- From brentn-at-gmail.com Tue Aug 22 21:00:32 2006 16, 27 -- Received: from wx-out-0506.google.com (wx-out-0506.google.com [66.249.82.232]) 16, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7N20WNQ004147 16, 27 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 21:00:32 -0500 16, 27 -- Received: by wx-out-0506.google.com with SMTP id t11so2335081wxc 16, 27 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 19:00:32 -0700 (PDT) 16, 27 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 16, 27 -- s=beta; d=gmail.com; 16, 27 -- h=received:mime-version:in-reply-to:references:content-type:message-id:content-transfer-encoding:from:subject:date:to:x-mailer:sender; 16, 27 -- b=bwBnQXHztGvZI8EEIWyvov8Ee15ACIWwMiqFhBFGAYTv5MRUOo4JrMAV1cfQmQFYZRdFGFOou/m5yfDCMpap52a0d9rYgyqs37+s94L4mUAp8mGdNm6+YG3F9XXcSmpXwxQyVGHePVwbBJ8K8MoDgFiV1y5lUy8IH4LRvAPpnnU= 16, 27 -- Received: by 10.70.90.18 with SMTP id n18mr12734220wxb; 16, 27 -- Tue, 22 Aug 2006 19:00:32 -0700 (PDT) 16, 27 -- Received: from ?192.168.123.152? ( [66.169.84.242]) 16, 27 -- by mx.gmail.com with ESMTP id 28sm254642wrl.2006.08.22.19.00.31; 16, 27 -- Tue, 22 Aug 2006 19:00:31 -0700 (PDT) 16, 27 -- Mime-Version: 1.0 (Apple Message framework v749.3) 16, 27 -- In-Reply-To: {200608221546.k7MFk6HI018268-at-ns.microscopy.com} 16, 27 -- References: {200608221546.k7MFk6HI018268-at-ns.microscopy.com} 16, 27 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 16, 27 -- Message-Id: {10DC6380-E144-47A9-B6DB-45921A38D022-at-freeshell.org} 16, 27 -- Content-Transfer-Encoding: 7bit 16, 27 -- From: Brent Neal {brentn-at-freeshell.org} 16, 27 -- Subject: Re: [Microscopy] Re: 16bit TIFF images in Photoshop 16, 27 -- Date: Tue, 22 Aug 2006 22:00:28 -0400 16, 27 -- To: microscopy-at-microscopy.com 16, 27 -- X-Mailer: Apple Mail (2.749.3) 16, 27 -- Sender: Brent Neal {brentn-at-gmail.com} ==============================End of - Headers==============================
Although this remark is right when one considers the auto-contrast option, I don't think you change the relative values of the pixels by adjusting the levels. As was described in another answer, reading this format in Photoshop compresses the values of the left side of the histogram. To recover a "normal" histogram, one has just to adjust the limits to exclude all the pixels with a 0 value which are outside of the histogram. I also forgot to point out that I work with ultrathin sections of biological material (ediffractometry is planned though). The requirements in other fields may be very different. Quantitative analysis are rare for me and when there are some to be done, it is usual area measurement, which does not depend on the individual value of each pixels.
Stephane
To Marc: Yes effectivement je parle francais
--- brentn-at-freeshell.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 } ---------------------------------------------------------------------------- } } } On Aug 22, 2006, at 11:46, voyles-at-engr.wisc.edu } wrote: } } } There is also a potentially serious drawback to } the Photoshop } } procedure } } you suggest. Photoshop discards the original } grey-scale values of the } } image when you use "Auto Levels", which breaks the } quantitative } } connection between the grey-scale value in the } image and the absolute } } number of electrons that struck the detector. } This will often make } } quantitative analysis of the electron image or } diffraction pattern } } impossible and makes it difficult to even judge } the relative average } } brightness of two different images to guess, for } example, the sample } } thickness. } } This is true. However, it is trivial to rectify the } problem that } he's having, which is that his system is saving } 12-bit TIFFs. Simply } use the Convolve function in Photoshop to multiply } the image by 16 } uniformly. This preserves the quantitative } connection while allowing } you to visually observe the image. (One notes that } there was a very } simple tutorial for this in Microscopy Today about } two years ago... *g*) } } } } Software developed for photographs and desktop } } publishing is not generally the best choice for } dealing with } } scientific } } data. } } I disagree with this statement. You can be } perfectly quantitative in } Photoshop, and in fact many people are (see all the } Fovea Pro users, } for example.) The issue at hand is one of needing } to understand what } one is doing to an image. But that issue is true no } matter what } software you're using. You can screw up data in } ImagePro just as } easily as you can in Photoshop. } } Brent } } } -- } Brent Neal } Research Physicist } Milliken Research Corporation } } } } } } } } ==============================Original } Headers============================== } 16, 27 -- From brentn-at-gmail.com Tue Aug 22 21:00:32 } 2006 } 16, 27 -- Received: from wx-out-0506.google.com } (wx-out-0506.google.com [66.249.82.232]) } 16, 27 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k7N20WNQ004147 } 16, 27 -- for {microscopy-at-microscopy.com} ; Tue, 22 } Aug 2006 21:00:32 -0500 } 16, 27 -- Received: by wx-out-0506.google.com with
I agree that cleaving is a great technique, very quick when you get the hang of it, and free of amorphous surface layers... but I've never managed to get a good cleave of something as ductile as pure Au, it always tears, delaminates or deforms. (There's still no such thing as an artefact-free specimen-prep method!) Since Geoff's colleages are looking at metal layers and already doing tripod polishing they might be able to get what they want with only a small change in approach. However if the materials are not too ductile, cleaving would be worth doing as well.
Just my two pennorths
Richard ________________________________________ Richard Beanland Materials Analysis Bookham Caswell Towcester Northants NN12 8EQ United Kingdom Tel. +44 1327 356362 Fax. +44 1327 356775 http://www.bookham.com ________________________________________
-----Original Message----- X-from: walck-at-southbaytech.com [mailto:walck-at-southbaytech.com] Sent: 23 August 2006 00:20 To: Richard Beanland
Thank you, Ron and Henk, for the plug. Sorry, I was on vacation or else I would have answered this.
I'd like to address the MicroCleave(TM) preparation at the end of this discussion so that it is closest to the disclaimer that I am going to put on it.
SDW on Microtoming: I've seen some images that Tom Malis collected of microtomed Si samples from different sources. True, you get a lot of mostly dust as Ron says, but apparently when you hit it using the right conditions, you can get little cracked pieces that compose the ribbon that have the film intact on them. If you are interested in only the film, you have a chance of getting what you need. However, for the most part, there are other methods for making Si cross sections that are much easier and much more likely for producing a usable sample.
SDW on Ion Milling: I have worked with thin Au-Ge-Ni multi-layers on GaAs in the past and you do have to worry about heating the sample as you indicate. You can use superglue in place of wax; you just have to be patient to get rid of the glue. If you ion mill your sample, you really have to worry about heating the sample during ion milling. The answer there is to use low angle, low energy, and it wouldn't hurt to cool the sample with a liquid nitrogen cooled stage. Since it is a competitor's product, I will not mention the name, but there is one popular ion mill on the market that you have to be particularly careful with. This ion mill does not have a sample holder that has a large heat sink and has a relatively poor thermal path through the holder. If you have this type of ion mill, you must significantly de-tune the ion guns or you can heat your samples very high even at low milling angles. Unfortunately, this "de-tuning" by lowering the ion energy and limiting the ion beam current will significantly impact the ion milling rate. I have melted glass cross sections with this ion mill. I have used other ion mills with the glass and for the most part, they are better in terms of not heating the glass samples simply because the stages dissipate the heat better, but you still have to be careful. In all of the conventional ion mills, i.e. non low-energy types, the beam current drops to essentially zero below about 3 kV. There are a number of studies that have been done concerning the heating of samples in ion mills. You can contact me off-line to talk about the low energy guns in the ion mills that we sell.
SDW on MicroCleaving: OK, let me sound off for the MicroCleave(TM), also known as the Small Angle Cleavage Technique. (You can even consider this coming from Igor Slobinsky and Dr. Cleavinstein so maybe I won't get in trouble for being too commercial.) I thought that this technique is the neatest thing since sliced bread (or sliced samples) when I started using it and I still do. This technique produces the best cross section samples of any available technique because there is simply no ion damage produced. In fact, a major disadvantage of these types of samples is that if the samples are fully crystalline and you are using a field emission gun equipped microscope, then there is no amorphous regions to help the microscopist focus and stigmate with. I suspect that with the new C(s)-corrected machines, more and more people will be discovering this technique.
For the metallization samples, the MicroCleave(TM) technique will work as long as the layers are very thin. Generally, they should be less than 1000A. They also must be fully adherent to the substrate and to the other layers. For as-deposited Au-Ge-Ni on GaAs samples with e-beam evaporated Ge layers that I mentioned above, I found that the Ge layers were weak and would split within the layer during the cleave. I've never looked at Ge layers put down by other methods, so I don't know whether that is a universal trait of Ge layers or not. If we heat treated them to alloy the layers, I had no such problem. Even a slight anneal where there was no apparent mixing of the layer materials would help prevent the Ge layer from splitting. I suspect that we might have been getting a little densification with the slight anneal. What happens with these samples during the procedure is that the thinnest parts of the samples would consist of any layers below the Ge and about half the Ge layer. Further back, all of the layers would be intact, but thicker. I can send a copy of a presentation on the MicroCleave technique that John McCaffrey and I did in "Cleave-Land" at an M&M meeting in 1997 to anyone that wants one. There are two places in the presentation that addresses temperature sensitive samples. First, as you point out, use superglue and then wait for the superglue to dissolve when finished instead of using the low temperature melting wax. The other place is that for the mounting epoxy, you need a viscous and slow setting epoxy to mount the samples onto the specially bent Veco 2x1 tabbed grids. Normally, Epoxy Technology's H-22 silver filled epoxy is used, but that requires heat to set. The 90 minute Super strength type of two part epoxies work well, but you have to give them a full day to set up. They are also nonconductive, but that didn't seem to be much of a problem. I've tried a number of different brands, and they seemed to work, but they have to be the 90 minute working time ones. Other epoxies flowed too much and did not stay in position to hold the sample. I also experimented with fillers for the epoxy with no success at all. There is also a temperature spike when an epoxy cures. When I used this on temperature sensitive samples, I tried to minimize the amount of epoxy on the grids so that the grid could act as a heat sink for the exothermic reaction and the samples would experience the minimum of a thermal excursion.
One other point of caution for about Si samples prepared this way. The samples can become very very thin and basically disappear into nothingness at the edges. I had a pulsed laser deposited diamond like carbon film that was deposited onto silicon and prepared this way that was too thin at the edges to do electron energy loss spectroscopy on and get sufficient signal! That's thin! What it means is that the samples prepared this way can be heated by a focused beam because there isn't enough thermal path to extract heat from the exposed area.
Here is a list of the references that I have. The "Update" paper by John and me also has an accompanying presentation that I can send to people in a PDF format that I can send to anyone that is interested. This presentation and the MRS paper by us have a pictorial outline of the technique in it. There are a lot of hands-on tips and tricks and details that we tried to put into the paper so that someone could try the technique without reinventing the wheel.
1) "Small Angle Cleavage of Semiconductors for TEM", J.P. McCaffrey, Ultramicroscopy 38, 1991, pp:149-157 2) "TEM Samples of Semiconductors Prepared by a Small Angle Cleavage Technique", J.P. McCaffrey, Mat. Res. Soc. Symp. Proc., Vol.254, 1992, pp:109-120 3) John P. McCaffrey, Microscopy Research and Technique, 24, (1993) 180. 4) "A Simplified Method for Modifying TEM Copper Grids For Use with the Small Angle Cleavage Technique", Scott D. Walck, Microscopy Today, (96-4), (1996). 5) The Small Angle Cleavage Technique Applied to Coatings and Thin Films, S. D. Walck and J. P. McCaffrey, Thin Solid Films, 308-309, (1997) 399. 6) "The Small Angle Cleavage Technique: An Update", Scott D. Walck, John P. McCaffrey, Mat.Res.Soc.Symp.Proc., Vol.480,1997,pp:149-170
OK, here is the disclaimer that I hope will keep me out of trouble. South Bay Technology Inc. makes and sells the Microcleave(TM) Kit. SBT also sells the Technoorg-Linda line of ion mills with the low energy ion gun.
-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: Tuesday, August 22, 2006 11:20 AM To: Walck-at-SouthBayTech.com
Geoff,
No you can't "cut" Si with a diamond knife. Si cleaves very easily. When the knife touches the Si, the Si cleaves along one of its preferred {111} crystallographic cleaving planes. That might be fine if you orient the Si such that a Si {111} crystallographic direction is perpendicular to the block face. Then if you are VERY lucky, you MIGHT get a cleaved piece of Si oriented with {111} faces (at the risk of your knife). Any article that I've seen that claims to have microtomed Si has shown lots of pictures of tiny {111} Si sections.
Assuming that you are successful, so what! There is no, or very little interest, in anything in the semiconductor world or in potentially oriented thin films deposited on Si, that is best displayed on top of a Si cross section with a {111} block face orientation. The angles are wrong.
Si wafers in semiconductor processing are almost all with (or very close to) a Si {001} direction perpendicular to the Si wafer surface. Devices fabricated in, or patterns deposited on, these wafers are not randomly oriented. They are almost always aligned with {011} planes perpendicular to the Si surface, i.e. the x, y orientations of the devices and patterns are parallel to {011} directions in the Si surface. Cross sections of interest for TEM or SEM analysis are oriented such that the microscope looks down on a {011} "block face." If you orient the specimen's block face so as to attempt to cut {011} cross sections, the diamond knife will touch the Si and immediately propagate a cleave down the nearest {111} plane despite the orientation of the block. As the specimen in the block is constrained by the epoxy holding it, the result is a flake of Si dust from just the surface. When the knife comes around again you get more dust.
Using a microtome to make Si {011}cross sections is like using a screw driver to drive a nail.
Instead of fighting the fact that Si cleaves, why not exploit this fact and prepare really great thin specimens by the microcleaving technique pioneered by McCaffrey and Igor. I'll copy them and invite them to expand on this suggestion. They'll point out that you can make nice thin sections of thin films on Si with a minimum of tools and at room temperature. There is a videotape that shows how to do it.
Ron Anderson
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 } ---------------------------------------------------------------------- } ------ } } I do not know exactly what I can divulge, but they have tried tripod polishing, and ion-milling but I am told they elevate the temperature and the layers need to be prepped and kept at room temps until imaging. } } Does that say enough? I still feel it's a bit irrelevant to the initial question. } } Is it possible to section a Si wafer? Am I correct in being skeptical? Or should I just kill a $4k knife or try an ancient already messed up and questionably anchored diamond in the attempt? } } I just figured someone out there might be able to share a bit of practical experience. } } Thanks! } } Geoff Williams } Leduc Bioimaging Facility Manager } Brown University } } http://www.brown.edu/Facilities/Leduc_Bioimaging_Facility/ } } } } ==============================Original } Headers============================== } 8, 29 -- From Geoffrey_Williams-at-brown.edu Tue Aug 22 10:59:55 2006 8, } 29 -- Received: from draco.services.brown.edu (draco.services.brown.edu [128.148.106.172]) } 8, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MFxtQK026422 } 8, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:59:55 -0500 } 8, 29 -- Received: from ex-gateway1-out.AD.Brown.Edu (ex-gateway1.ad.brown.edu [128.148.21.51]) } 8, 29 -- by draco.services.brown.edu (Switch-3.1.8/Switch-3.1.7/) with ESMTP id k7MFxs7W006189 } 8, 29 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 11:59:54 -0400 (EDT) } 8, 29 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway1-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); } 8, 29 -- Tue, 22 Aug 2006 11:59:54 -0400 } 8, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 29 } -- Content-class: urn:content-classes:message 8, 29 -- MIME-Version: } 1.0 8, 29 -- Content-Type: text/plain; } 8, 29 -- charset="iso-8859-1" } 8, 29 -- Subject: Sectioning Si, clarification 8, 29 -- Date: Tue, 22 } Aug 2006 11:59:53 -0400 8, 29 -- Message-ID: } {A1A84D541C161C4C988B4E0FB38F5A0F047F6793-at-MAIL1.AD.Brown.Edu} } 8, 29 -- X-MS-Has-Attach: } 8, 29 -- X-MS-TNEF-Correlator: } 8, 29 -- Thread-Topic: Sectioning Si, clarification 8, 29 -- } Thread-Index: AcbGBAF0V914xLqeQPmnA7STgTxe2Q== 8, 29 -- From: } "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 8, 29 -- To: } {Microscopy-at-microscopy.com} 8, 29 -- X-OriginalArrivalTime: 22 Aug } 2006 15:59:54.0245 (UTC) FILETIME=[02015B50:01C6C604] 8, 29 -- } X-Brown-Proofpoint: Not Infected 8, 29 -- X-Proofpoint-Spam-Details: } rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 } reason=safe engine=3.1.0-0608080000 definitions=main-0608220003 8, 29 } -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam } policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe } engine=3.1.0-0608080000 definitions=main-0608220003 8, 29 -- } Content-Transfer-Encoding: 8bit 8, 29 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id k7MFxtQK026422 } ==============================End of - } Headers============================== } } }
==============================Original Headers============================== 11, 21 -- From randerson20-at-tampabay.rr.com Tue Aug 22 13:16:47 2006 11, 21 -- Received: from ms-smtp-06.tampabay.rr.com (ms-smtp-06.tampabay.rr.com [65.32.5.136]) 11, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MIGj91015481 11, 21 -- for {Microscopy-at-Microscopy.Com} ; Tue, 22 Aug 2006 13:16:46 -0500 11, 21 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 11, 21 -- by ms-smtp-06.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k7MIGVGD023421; 11, 21 -- Tue, 22 Aug 2006 14:16:37 -0400 (EDT) 11, 21 -- Message-ID: {44EB49FD.2020607-at-tampabay.rr.com} 11, 21 -- Date: Tue, 22 Aug 2006 14:16:29 -0400 11, 21 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 11, 21 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 11, 21 -- MIME-Version: 1.0 11, 21 -- To: Geoffrey_Williams-at-brown.edu, Listserver {Microscopy-at-Microscopy.Com} , 11, 21 -- John McCaffrey {mccaffrey2002-at-rogers.com} , 11, 21 -- Scott Walck {walck-at-southbaytech.com} 11, 21 -- Subject: Re: [Microscopy] Sectioning Si, clarification 11, 21 -- References: {200608221600.k7MG02nF026627-at-ns.microscopy.com} 11, 21 -- In-Reply-To: {200608221600.k7MG02nF026627-at-ns.microscopy.com} 11, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 11, 21 -- Content-Transfer-Encoding: 7bit 11, 21 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
==============================Original Headers============================== 29, 22 -- From walck-at-southbaytech.com Tue Aug 22 18:18:57 2006 29, 22 -- Received: from ylpvm43.prodigy.net (ylpvm43-ext.prodigy.net [207.115.57.74]) 29, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MNIv4K025370 29, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 18:18:57 -0500 29, 22 -- X-ORBL: [64.169.193.90] 29, 22 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 29, 22 -- by ylpvm43.prodigy.net (8.13.7 out spool5000 dk/8.13.7) with ESMTP id k7MNIsbi030444; 29, 22 -- Tue, 22 Aug 2006 19:18:55 -0400 29, 22 -- From: "Scott Walck" {walck-at-southbaytech.com} 29, 22 -- To: {Microscopy-at-microscopy.com} 29, 22 -- Cc: {randerson20-at-tampabay.rr.com} , "Henk Colijn" {colijn.1-at-osu.edu} 29, 22 -- Subject: RE: [Microscopy] Re: Sectioning Si, clarification 29, 22 -- Date: Tue, 22 Aug 2006 16:19:36 -0700 29, 22 -- Message-ID: {00a301c6c641$6f4255b0$7801a8c0-at-dynamicbl8uno3} 29, 22 -- MIME-Version: 1.0 29, 22 -- Content-Type: text/plain; 29, 22 -- charset="us-ascii" 29, 22 -- Content-Transfer-Encoding: 7bit 29, 22 -- X-Mailer: Microsoft Office Outlook 11 29, 22 -- In-Reply-To: {200608221820.k7MIKOHm021597-at-ns.microscopy.com} 29, 22 -- Thread-Index: AcbGF6KHgJf2sp57RhCWrZ22d+K1vQACaacw 29, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
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==============================Original Headers============================== 40, 32 -- From richard.beanland-at-bookham.com Wed Aug 23 01:45:46 2006 40, 32 -- Received: from mail80.messagelabs.com (mail80.messagelabs.com [195.245.230.163]) 40, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7N6jjBN028944 40, 32 -- for {microscopy-at-microscopy.com} ; Wed, 23 Aug 2006 01:45:46 -0500 40, 32 -- X-VirusChecked: Checked 40, 32 -- X-Env-Sender: richard.beanland-at-bookham.com 40, 32 -- X-Msg-Ref: server-6.tower-80.messagelabs.com!1156315544!39764504!1 40, 32 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 40, 32 -- X-Originating-IP: [213.249.209.179] 40, 32 -- Received: (qmail 26644 invoked from network); 23 Aug 2006 06:45:44 -0000 40, 32 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 40, 32 -- by server-6.tower-80.messagelabs.com with SMTP; 23 Aug 2006 06:45:44 -0000 40, 32 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 40, 32 -- Wed, 23 Aug 2006 07:48:05 +0100 40, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 40, 32 -- Content-class: urn:content-classes:message 40, 32 -- MIME-Version: 1.0 40, 32 -- Content-Type: text/plain; 40, 32 -- charset="us-ascii" 40, 32 -- Subject: RE: [Microscopy] Sectioning Si, clarification 40, 32 -- Date: Wed, 23 Aug 2006 07:48:08 +0100 40, 32 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E1BA348-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 40, 32 -- X-MS-Has-Attach: 40, 32 -- X-MS-TNEF-Correlator: 40, 32 -- Thread-Topic: [Microscopy] Sectioning Si, clarification 40, 32 -- Thread-Index: AcbGQicZdxUSD9c8TWu0CVtQfyaqrQAOyIfQ 40, 32 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 40, 32 -- To: {walck-at-southbaytech.com} 40, 32 -- Cc: {microscopy-at-microscopy.com} 40, 32 -- X-OriginalArrivalTime: 23 Aug 2006 06:48:05.0595 (UTC) FILETIME=[16200AB0:01C6C680] 40, 32 -- Content-Transfer-Encoding: 8bit 40, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7N6jjBN028944 ==============================End of - Headers==============================
Even cheapo £36 Roxio Photosuite is actually better than Photoshop for creating montages (not difficult as Photoshop CS2’s File, Automate, Photomerge is actually very poor). I assume by 'montage' you mean photo-stitching a series of 500 consecutive raster scan images into one rather large one (e.g. a panorama view). Photo-stitching software does normally require an 'overlap' though between the images to work well (otherwise I'm sure you could use Photoshop semi-automatically via it's 'actions' macros to cut and paste images into a large image and then use Edit, Transform to manually align all the layers in Photoshop).
Naturally Roxio Photosuite doesn’t hold a candle to Photoshop for most other image editing things though. We have very expensive photo-stitching software here for our new TEM Gatan digital camera system (that even comes with it’s own PC work-stations).
Very sophisticated photo-stitching software is available for around £300 though (e.g. RealViz Stitcher Unlimited) http://www.pcw.co.uk/personal-computer-world/software/2162780/realviz-stitch er-unlimited
Plus there’s RealViz Express at around £70: http://www.pcw.co.uk/personal-computer-world/software/2137457/realviz-stitch er-express “This is more expensive than other budget panorama applications, but Stitched Express is every bit as easy to use and its professional features make them look like toys by comparison”.
Try, buy and download at http://stitcher.realviz.com.
For home use (stitching SLR digital images of scenes) I find that software that came with our Camera’s (Canon and Olympus) performs far far better at Photo-stitching than PhotoShop’s PhotoMerge. In particular Olympus Master Pro software (a £20 upgrade from the basic software that comes for with the camera is needed for photo-stitching) seems to perform very well, but I’ve never tried it with scientific images (yet). SEM images are easier to photostitch as they lack complex colour information. The main trick is to take all colour images at the same exposure etc.., probably setting via the brightest area first, so that all images have similar colour balance and brightness/contrast.
Photoshop is great for subsequently blending in any really obvious joins, e.g. by cutting & pasting, and blurring (and don’t forget things like Cut&paste, Edit, Transform, Distort to line up broken lines etc.).
Five hundred images is a lot to deal with in one go, so as mentioned by others, doing them in many smaller series and stitching the results will probably be needed.
There's a great deal of photographic help on the web, e.g. If you want to create an artistic blended 'collage' instead, see http://www.thinkdan.com/tutorials/photoshop/montage.pdf http://www.dwphotoshop.com/photoshop/MontageTutorial.php http://graphicssoft.about.com/od/photoshoptutorialscollage
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: jds451-at-psu.edu [mailto:jds451-at-psu.edu] Sent: 20 August 2006 16:02 To: keith.morris-at-ucl.ac.uk
Hi all, I have a friend who needs to make a montage of a large number of optical micrographs. He has about 500 images. Can anybody suggest a PC software package that is easy to use? He is currently using Photoshop. Freeware or something that can be paid for and downloaded would be especially useful. Thanks for any suggestions,
Jennifer Ray Sloppy Materials Science & Engineering, Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 3, 17 -- From jds451-at-psu.edu Sun Aug 20 09:51:57 2006 3, 17 -- Received: from webmail19.cac.psu.edu (webmail19.cac.psu.edu [128.118.1.205]) 3, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7KEpvZI004541 3, 17 -- for {microscopy-at-microscopy.com} ; Sun, 20 Aug 2006 09:51:57 -0500 3, 17 -- Received: (from webmail-at-localhost) 3, 17 -- by webmail19.cac.psu.edu (8.9.3/8.9.0) id KAA20369; 3, 17 -- Sun, 20 Aug 2006 10:51:56 -0400 (EDT) 3, 17 -- Date: Sun, 20 Aug 2006 10:51:56 -0400 (EDT) 3, 17 -- Message-Id: {200608201451.KAA20369-at-webmail19.cac.psu.edu} 3, 17 -- To: microscopy-at-microscopy.com 3, 17 -- Subject: Software: Montage 3, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} 3, 17 -- X-Mailer: Penn State WebMail 3, 17 -- X-Sender: jds451 3, 17 -- X-Originating-IP: 207.255.156.167 3, 17 -- MIME-Version: 1.0 3, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
==============================Original Headers============================== 25, 27 -- From keith.morris-at-ucl.ac.uk Wed Aug 23 07:23:49 2006 25, 27 -- Received: from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 25, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7NCNn4V011924 25, 27 -- for {Microscopy-at-microscopy.com} ; Wed, 23 Aug 2006 07:23:49 -0500 25, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 25, 27 -- by vscani-e.ucl.ac.uk with esmtp (Exim 4.60) 25, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 25, 27 -- id 1GFrlk-0001YZ-Aq; Wed, 23 Aug 2006 13:23:43 +0100 25, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 25, 27 -- To: {jds451-at-psu.edu} 25, 27 -- Cc: {Microscopy-at-microscopy.com} 25, 27 -- Subject: RE: [Microscopy] Software: Montage 25, 27 -- Date: Wed, 23 Aug 2006 13:23:37 +0100 25, 27 -- Message-ID: {006201c6c6ae$f653bd10$7b865290-at-keithhigrade} 25, 27 -- MIME-Version: 1.0 25, 27 -- Content-Type: text/plain; 25, 27 -- charset="iso-8859-1" 25, 27 -- X-Mailer: Microsoft Office Outlook 11 25, 27 -- Thread-Index: AcbEaYzU7D+4C/dCRB6dovc7UHj/zgCNXHhw 25, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 25, 27 -- In-Reply-To: {200608201501.k7KF1VQf012051-at-ns.microscopy.com} 25, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 25, 27 -- X-UCL-MailScanner: Found to be clean 25, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 25, 27 -- X-Spam-Status: No 25, 27 -- Content-Transfer-Encoding: 8bit 25, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7NCNn4V011924 ==============================End of - Headers==============================
I would like to know if anyone of you already have had success in staining with the PAS (Schiff) on Epon sections without previous etching of Epon. On internet I found one reference which says that “may” work. The purpose is to stain glycogen in cell embedded in Epon, sections approx 200-500 nm thick. If you know any other reaction which works on these sections I take too :-D
regards,
Stephane
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==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Wed Aug 23 08:12:39 2006 6, 18 -- Received: from web37411.mail.mud.yahoo.com (web37411.mail.mud.yahoo.com [209.191.91.143]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7NDCdPM022952 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 23 Aug 2006 08:12:39 -0500 6, 18 -- Received: (qmail 51108 invoked by uid 60001); 23 Aug 2006 13:12:39 -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=wX3QknE8b/KkweiobnQ7eyOPkwww9GL3YFyVtSLSTQRI/vBU7lBsoCYM9XHtz48/ydVTf6aZr2VHE/fMl/AnjdDQL4c6DwNsMklmN9/492rFpMfzNy5XETnJxWcxRsfCqh3QoMHfp/zsH4lM2tdnMze5lhNr0Jhy3fpySqYYbL4= ; 6, 18 -- Message-ID: {20060823131239.51106.qmail-at-web37411.mail.mud.yahoo.com} 6, 18 -- Received: from [80.122.101.102] by web37411.mail.mud.yahoo.com via HTTP; Wed, 23 Aug 2006 06:12:39 PDT 6, 18 -- Date: Wed, 23 Aug 2006 06:12:39 -0700 (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: PAS on Epon sections 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==============================
Some years ago, I did PAS staining of Epon sections (0.5-1.0 micron thick). There is a good reference explaining the procedure: Schroeder HE, Rossinsky K, Müller W (1980) An established routine method for differential staining of epoxy-embedded tissue sections. Microscopica Acta 83,111-116. Hope this helps. Best regards, Dieter -- Dieter D. Bosshardt, Ph.D. University of Bern School of Dental Medicine Department of Periodontology & Fixed Prosthodontics Post Box 64 Freiburgstrasse 7 CH-3010 Bern 10 Switzerland http://dent.unibe.ch Phone: +41-31-632 86 05 Fax: +41-31-632 49 31 mailto:dieter.bosshardt-at-zmk.unibe.ch
==============================Original Headers============================== 2, 28 -- From dieter.bosshardt-at-zmk.unibe.ch Wed Aug 23 08:28:42 2006 2, 28 -- Received: from mailhub03.unibe.ch (mailhub.unibe.ch [130.92.9.70]) 2, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7NDSgJ6000805 2, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 23 Aug 2006 08:28:42 -0500 2, 28 -- Received: from localhost (scanhub02.unibe.ch [130.92.5.62]) 2, 28 -- by mailhub03.unibe.ch (Postfix) with ESMTP id DC60F4AE68 2, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 23 Aug 2006 15:28:41 +0200 (CEST) 2, 28 -- X-Virus-checked: by University of Berne 2, 28 -- Received: from mailhub03.unibe.ch ([130.92.9.70]) 2, 28 -- by localhost (scanhub02.unibe.ch [130.92.5.62]) (amavisd-new, port 10024) 2, 28 -- with LMTP id 6BNmPAgsjaQA for {Microscopy-at-microscopy.com} ; 2, 28 -- Wed, 23 Aug 2006 15:28:39 +0200 (CEST) 2, 28 -- Received: from ubecx01 (ubecx01.unibe.ch [130.92.6.40]) 2, 28 -- by mailhub03.unibe.ch (Postfix) with ESMTP id 50A094BEAE 2, 28 -- for {Microscopy-at-microscopy.com} ; Wed, 23 Aug 2006 15:28:40 +0200 (CEST) 2, 28 -- Received: from [130.92.198.55] (zmkparo2.unibe.ch [130.92.198.55]) 2, 28 -- by ubecx01.unibe.ch (PMDF V5.2-32 #42481) 2, 28 -- with ESMTP id {0J4G009IIDFQSX-at-ubecx01.unibe.ch} for Microscopy-at-microscopy.com; 2, 28 -- Wed, 23 Aug 2006 15:28:39 +0200 (MEST) 2, 28 -- Date: Wed, 23 Aug 2006 15:28:37 +0200 2, 28 -- From: Dieter Bosshardt {dieter.bosshardt-at-zmk.unibe.ch} 2, 28 -- Subject: PAS staining of Epon sections 2, 28 -- To: Microscopy-at-microscopy.com 2, 28 -- Message-id: {a06230900c112074edf0a-at-[130.92.198.55]} 2, 28 -- MIME-version: 1.0 2, 28 -- Content-type: text/plain; charset="iso-8859-1" ; format="flowed" 2, 28 -- Content-Transfer-Encoding: 8bit 2, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7NDSgJ6000805 ==============================End of - Headers==============================
I really can't see the problem. We can only resolve 191 grey levels and 8-bit is 256 grey levels - so in practice we can't see the difference between 8-bit, 12-bit or 16-bit B&W, except in the case of file size when true 16-bit is quite massive. Likewise I defy anyone to be able to tell the difference by eye between colour images in 24-bit, 36-bit and 48-bit (other than file storage size). Photoshop should be happy with true 16-bit B&W (32,768 grey levels) if you are.
Anyone wishing to view images from say 12-bit OpenLabs or Bio-Rad Pic files would normally export 8-bit tiff's from the imaging software, retaining the 12-bit master images for any image analysis packages that can read them. This is very quick and easy to do, with export being largely automatic. In practice you would only use 12-bit for determining image 'brightness' via pixel intensities (although even here 256 grey levels is actually less complicated than having 4,096 grey levels). Biological variation, labelling efficiency, bleaching and imaging hardware drift make any increased 12-bit measurement 'accuracy' a little meaningless anyway. Naturally you don't want to mess about with contrast/brightness/gamma for quantitative image intensity comparisons. Thus 'recovering' image information from 12-bit B&W image in Photoshop is dubious (and very time-consuming) for intensity comparisons (but absolutely fine if you just want to see the image quickly - which I think is all Stephane was saying). For any other measurements such as count, area and perimeter, bit depth is irrelevant as you are working with the binary (only 1-bit) thresholded overlay image.
Likewise Photoshop can't read 12-bit or even 8-bit multi-tiff z stacks (you just get the first image as naturally it expects a 'photograph' not a video type sequence), whereas image analysis packages like ImageJ and MetaMorph can read multi-tiff (as they have to for time-lapse & z-stack sequences).
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: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: 22 August 2006 16:27 To: keith.morris-at-ucl.ac.uk
Dear listers,
I think I have an easy solution for a common problem and I just wanted to share it. I am sorry if I bring nothing new, but perhaps it really helps. At least it cannot harm :-)
When working with my TEMcam Megaview III and the software AnalySIS (no personal interest blablabla...) the pictures are saved in 16bit TIFF format. This is a problem when you want to work with them because this format is not recognized by most image processing software (This problem is so common that it appears in the FAQ of AnalySIS). However when this format is loaded in Adobe Photoshop (no personal....) the picture comes completely dark. A rather simple solution to recover your picture is to use the "auto contrast" and/or "auto level" option and your nice picture appears in all its splendor right before your eyes!
Hope this was helpful. Have a nice day (for the american), a nice sleep (for the australian) or a nice evening (for the european).
Stephane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Tue Aug 22 10:22:55 2006 7, 18 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.91.142]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7MFMtJ5027602 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 22 Aug 2006 10:22:55 -0500 7, 18 -- Received: (qmail 21837 invoked by uid 60001); 22 Aug 2006 15:22:55 -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=FP2WVswviXCCARHnXwNLh4HVu9wy6B+uLcz1srxmBXfOK7AOK1yXsSzEW/ImrlWN4trgP7WdCa hgLGDn8k2mb/DEqR/6OtgyruYcPY4CSc5LB0WGkWkbw38+DvR5OuvAvRI51DYVL4elyoXJJaPVjo Bn3cOnsVMP+LHoCAm6+yA= ; 7, 18 -- Message-ID: {20060822152255.21835.qmail-at-web37410.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Tue, 22 Aug 2006 08:22:55 PDT 7, 18 -- Date: Tue, 22 Aug 2006 08:22:55 -0700 (PDT) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: 16bit TIFF images in Photoshop 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============================== 24, 26 -- From keith.morris-at-ucl.ac.uk Wed Aug 23 08:56:53 2006 24, 26 -- Received: from vscani-a.ucl.ac.uk (vscani-a.ucl.ac.uk [144.82.108.29]) 24, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7NDuqP9011817 24, 26 -- for {Microscopy-at-microscopy.com} ; Wed, 23 Aug 2006 08:56:53 -0500 24, 26 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 24, 26 -- by vscani-a.ucl.ac.uk with esmtp (Exim 4.60) 24, 26 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 24, 26 -- id 1GFtDt-0005hQ-Fl 24, 26 -- for Microscopy-at-microscopy.com; Wed, 23 Aug 2006 14:56:49 +0100 24, 26 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 24, 26 -- To: {Microscopy-at-microscopy.com} 24, 26 -- Subject: RE: [Microscopy] 16bit TIFF images in Photoshop 24, 26 -- Date: Wed, 23 Aug 2006 14:56:46 +0100 24, 26 -- Message-ID: {006c01c6c6bb$f9231dd0$7b865290-at-keithhigrade} 24, 26 -- MIME-Version: 1.0 24, 26 -- Content-Type: text/plain; 24, 26 -- charset="us-ascii" 24, 26 -- Content-Transfer-Encoding: 7bit 24, 26 -- X-Mailer: Microsoft Office Outlook 11 24, 26 -- Thread-Index: AcbF/3NlrBXInUHBQd+pwicfoKXjxQAsdZnw 24, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 24, 26 -- In-Reply-To: {200608221527.k7MFRCPC002191-at-ns.microscopy.com} 24, 26 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 24, 26 -- X-UCL-MailScanner: Found to be clean 24, 26 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 24, 26 -- X-Spam-Status: No ==============================End of - Headers==============================
1. Iodine would be useful. 1 g iodine 2 g Potassium iodide 100 ml Distilled water Store in colored glass as stock
Add 5 ml stock to 100 ml water for actual use
Raw amylose-containing starches - blue Raw amylopection-containing starches - reddish Chemically modified starches - yellow-brown Pregelatinixed/cooked - reddish and swollen
Dextrins - blue-purple Proteins in crysosections - yellow
2. Also Trypan blue to distinguished damaged starch:
0.25 g of dye 100 ml Distilled Water Leave on for 1-3 min, blot off excess
Damaged - blue Bit of Lost structure - pale blue Undamaged - unstained
Protein - lighter blue Lignified cellulose - Dark blue Cellulose (plant cell walls)- Pale blue
Mold (chitinous) - blue
3. As far as Polarization.
In cross polars fresh starch will show well as a malteese cross, if it has been cooked it will not. Embedding mediums to one degree or another - depending on media birefrigence and strain, can give a background polarization that can be countered at the right rotation.
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
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Email: twigg-at-estd.nrl.navy.mil Name: Mark Twigg
Organization: MSA
Title-Subject: [Filtered] Micro Saw
Question: I am interested in purchasing a diamond saw for cutting up XTEM samples. I need something that can cut small pieces of SiC, sapphire, or silicon wafers--espeically for H-cut FIB. My technician recently retired and he used to cut up my samples using a gigantic saw about the size of the band saw that I used in high school wood shop forty years ago. Although I could use this old saw, it is really gross overkill for what I really need. I would be interested in hearing about the capabilities and prices of smaller saws. I see that South Bay, for example, has a MicroSaw (TL-MS1) on the market. Does anybody out there have any experience with this saw or similar saws?
A student here is working on a project about magnification for children. He's starting with a wooden ruler and taking pictures with different microscopes. I can get him an image of wood up to 300,000 with my SEM. Does anyone have a picture of wood or any other plant material (at high mags, it doesn't make any difference for this) with SEM over 300,000x or with AFM? Or any other technique that will demonstrate how high a magnification that can be achieved with microscopy.
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 Thu Aug 24 16:38:15 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 k7OLcFgq032049 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 24 Aug 2006 16:38:15 -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 k7OLcBOP027732 5, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 24 Aug 2006 11:38:12 -1000 (HST) 5, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k7OLcA3s027729 5, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 24 Aug 2006 11:38:11 -1000 (HST) 5, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 5, 19 -- Date: Thu, 24 Aug 2006 11:38:10 -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: AFM/SEM of wood/plant material 5, 19 -- Message-ID: {Pine.GSO.4.21.0608241134260.27655-100000-at-halia} 5, 19 -- MIME-Version: 1.0 5, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
Dear Tina, We have a nice AFM image of a wood pulp fiber which shows the cellulose microfibrils (ca. 20-30 nm wide) as well as patches of residual lignin on the surface. Aside from magnification, this illustrates that wood is a composite material. Please see http://www.asmicro.com/Applications/phase.htm
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: tina-at-pbrc.hawaii.edu To: donc-at-asmicro.com Sent: Thursday, August 24, 2006 5:43 PM Subject: [a] [Microscopy] AFM/SEM of wood/plant material
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Hi, All-
A student here is working on a project about magnification for children. He's starting with a wooden ruler and taking pictures with different microscopes. I can get him an image of wood up to 300,000 with my SEM. Does anyone have a picture of wood or any other plant material (at high mags, it doesn't make any difference for this) with SEM over 300,000x or with AFM? Or any other technique that will demonstrate how high a magnification that can be achieved with microscopy.
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*
I need to do an experiment in Arabidopsis to localize a GFP tagged protein in seed coat cells by immunogold. However, there have been a few discussions in our lab about the quality of the various anti-GFPs that are available to purchase, and we haven't been able to come to a consensus as to which one to use.
I was hoping that by asking on the listserve, I could get an idea of where people get their anti-GFPs and whether they were happy with them. I seem to remember this question coming up in the past, but I never saw the response.
The questions I have for the anti-GFP users out there is this:
1.) Where do you get your anti-GFP?
2.) Are there any caveats that you've found while using this particuliar type (ie. prefers low salt conditions, not very clean on westerns, etc.) that should be taken into consideration when using it?
3.) Is there a reference I could take a peek at where you successfully used this anti-GFP for immunogold?
4.) Have you tried any other anti-GFPs than the one you currently use?
Please respond offlist (i.e. directly to me) , and I will compile the answers and submit them back to the listserver
Thanks very much in advance, Robin
--------------------------------- Robin Young, M.Sc. PhD Candidate Dept of Botany, UBC 6270 University Blvd. Vancouver, BC, Canada V6T 1Z4 fax: 604-822-6089
==============================Original Headers============================== 12, 27 -- From youngre-at-interchange.ubc.ca Thu Aug 24 19:16:55 2006 12, 27 -- Received: from mta5.mail-relay.ubc.ca (mta5.mail-relay.ubc.ca [137.82.45.10]) 12, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7P0GsW0022272 12, 27 -- for {microscopy-at-microscopy.com} ; Thu, 24 Aug 2006 19:16:54 -0500 12, 27 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 12, 27 -- by mta5.mail-relay.ubc.ca (8.12.11.20060308/8.12.11) with ESMTP id k7P0GrkW016604 12, 27 -- for {microscopy-at-microscopy.com} ; Thu, 24 Aug 2006 17:16:53 -0700 (PDT) 12, 27 -- (envelope-from youngre-at-interchange.ubc.ca) 12, 27 -- Received: from [137.82.136.201] (robin.botany.ubc.ca [137.82.136.201]) 12, 27 -- by smtp.interchange.ubc.ca 12, 27 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 12, 27 -- with ESMTPSA id {0J4J009E72453V-at-smtp.interchange.ubc.ca} for 12, 27 -- microscopy-at-microscopy.com; Thu, 24 Aug 2006 17:16:53 -0700 (PDT) 12, 27 -- Date: Thu, 24 Aug 2006 17:16:54 -0700 12, 27 -- From: Robin Young {youngre-at-interchange.ubc.ca} 12, 27 -- Subject: Anti-GFP for TEM? 12, 27 -- To: microscopy-at-microscopy.com 12, 27 -- Message-id: {88C5CD59-B3B5-4B8B-8F05-FF98DFA1F1BD-at-interchange.ubc.ca} 12, 27 -- MIME-version: 1.0 (Apple Message framework v752.2) 12, 27 -- X-Mailer: Apple Mail (2.752.2) 12, 27 -- Content-type: text/plain; format=flowed; delsp=yes; charset=US-ASCII 12, 27 -- Content-transfer-encoding: 7bit 12, 27 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.8.24.165442 12, 27 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 12, 27 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P1_5 0, __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 12, 27 -- X-Spam-Level: 12, 27 -- X-Spam-Flag: No ==============================End of - Headers==============================
We have have since à year for the same type of use a Well diamond wire saw, which works very good for what we need. Have a look at :
http://www.welldiamondwiresaws.com/
Our model is the 3242 from the Serie 3000. The mounting of the sample is a real "meccano", as we say in french, or "lego", to speak with a more international and today image. One can build very easy any holder type to mount simple or complicated samples to be cut. It can cut very thin sections, with a surface state which avoid the gross polishing steps. The wire is open and spooled on two threaded drums, which rotate and move, to wind and unwind it, mainening it allways at the same position in regard to the sample. If broken it can be changed very easy by the user.
And it's not a carpenter tool ! It makes h40*L40*l20 cm. Of coarse, Well sells bigger saws too !
We use it for TEM and SEM.
No financial interest, only a satisfied customer.
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============================== 13, 30 -- From jacques.faerber-at-ipcms.u-strasbg.fr Fri Aug 25 01:50:41 2006 13, 30 -- Received: from mailhost.u-strasbg.fr (mailhost.u-strasbg.fr [130.79.200.153]) 13, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7P6oeoj005477 13, 30 -- for {microscopy-at-microscopy.com} ; Fri, 25 Aug 2006 01:50:40 -0500 13, 30 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr [130.79.210.2]) 13, 30 -- by mailhost.u-strasbg.fr (8.13.6/jtpda-5.5pre1) with ESMTP id k7P6oZTA006221 13, 30 -- ; Fri, 25 Aug 2006 08:50:35 +0200 (CEST) 13, 30 -- Received: from [130.79.54.3] (odhinn.u-strasbg.fr [130.79.54.3]) 13, 30 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id 8789E10001BB; 13, 30 -- Fri, 25 Aug 2006 08:46:01 +0200 (CEST) 13, 30 -- Message-ID: {44EE9C81.2080403-at-ipcms.u-strasbg.fr} 13, 30 -- Date: Fri, 25 Aug 2006 08:45:21 +0200 13, 30 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} 13, 30 -- User-Agent: Mozilla Thunderbird 1.0.8 (X11/20060503) 13, 30 -- X-Accept-Language: fr, en 13, 30 -- MIME-Version: 1.0 13, 30 -- To: twigg-at-estd.nrl.navy.mil, Microscopy-at-microscopy.com 13, 30 -- Subject: Re: [Microscopy] viaWWW: Micro Saw 13, 30 -- References: {200608241413.k7OED2Q3023257-at-ns.microscopy.com} 13, 30 -- In-Reply-To: {200608241413.k7OED2Q3023257-at-ns.microscopy.com} 13, 30 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 30 -- Content-Transfer-Encoding: 8bit 13, 30 -- X-IPCMS-MailScanner: Found to be clean 13, 30 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr 13, 30 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-2.0.2 (mailhost.u-strasbg.fr [130.79.200.153]); Fri, 25 Aug 2006 08:50:35 +0200 (CEST) 13, 30 -- X-Virus-Scanned: ClamAV 0.88.4/1728/Fri Aug 25 07:55:58 2006 on mr3.u-strasbg.fr 13, 30 -- X-Virus-Status: Clean 13, 30 -- X-Spam-Status: No, score=-0.5 required=5.0 tests=AWL autolearn=disabled 13, 30 -- version=3.1.4 13, 30 -- X-Spam-Checker-Version: SpamAssassin 3.1.4 (2006-07-25) on mr3.u-strasbg.fr ==============================End of - Headers==============================
Hi Mark, The diamond wire saws are very expensive, although excellent. For ceramic materials we have also used a simple diamond wafering saw. You can get them from: BUEHLER, STRUERS, LECO, etc... In addition, we have found that hollow diamond-coated drill bits work very well for drilling out 2.8mm diameter rods from x-sections of ceramics, and even ceramic-metal interfaces. You can insert these rods into Cu tubes for subsequent x-section preparation, following the method developed by the people in Manfred Rֳ¼hle's group at MPI-Stuttgart.
Wayne
Prof. Wayne D. Kaplan Dept. of Materials Engineering Technion - Israel Institute of Technology Haifa 32000 Israel Tel/Fax: (972)-(0)4-829-4580 Cell: (972)-(0)50-7629377 kaplan-at-tx.technion.ac.il www.technion.ac.il/~kaplan
-----Original Message----- X-from: jacques.faerber-at-ipcms.u-strasbg.fr [mailto:jacques.faerber-at-ipcms.u-strasbg.fr] Sent: Friday, 25 August, 2006 8:53 AM To: Wayne D. Kaplan
Hi Mark
We have have since ׳ year for the same type of use a Well diamond wire saw, which works very good for what we need. Have a look at :
http://www.welldiamondwiresaws.com/
Our model is the 3242 from the Serie 3000. The mounting of the sample is a real "meccano", as we say in french, or "lego", to speak with a more international and today image. One can build very easy any holder type to mount simple or complicated samples to be cut. It can cut very thin sections, with a surface state which avoid the gross polishing steps. The wire is open and spooled on two threaded drums, which rotate and move, to wind and unwind it, mainening it allways at the same position in regard to the sample. If broken it can be changed very easy by the user.
And it's not a carpenter tool ! It makes h40*L40*l20 cm. Of coarse, Well sells bigger saws too !
We use it for TEM and SEM.
No financial interest, only a satisfied customer.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Mat׳™riaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
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==============================Original Headers============================== 23, 34 -- From kaplan-at-mt.technion.ac.il Fri Aug 25 03:17:17 2006 23, 34 -- Received: from mailgw1.technion.ac.il (mailgw1.technion.ac.il [132.68.238.34]) 23, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7P8HGe4016836 23, 34 -- for {Microscopy-at-microscopy.com} ; Fri, 25 Aug 2006 03:17:17 -0500 23, 34 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 23, 34 -- by mailgw1.technion.ac.il (Postfix) with ESMTP id CF161AC64D 23, 34 -- for {Microscopy-at-microscopy.com} ; Fri, 25 Aug 2006 11:15:50 +0300 (IDT) 23, 34 -- X-Virus-Scanned: by amavisd-new at technion.ac.il 23, 34 -- Received: from mailgw1.technion.ac.il ([127.0.0.1]) 23, 34 -- by localhost (mailgw1.technion.ac.il [127.0.0.1]) (amavisd-new, port 10024) 23, 34 -- with LMTP id Gqt03n3l2J3L for {Microscopy-at-microscopy.com} ; 23, 34 -- Fri, 25 Aug 2006 11:15:50 +0300 (IDT) 23, 34 -- Received: from td-cc-mail.cc.technion.ac.il (td-cc-mail.cc.technion.ac.il [132.68.3.30]) 23, 34 -- by mailgw1.technion.ac.il (Postfix) with ESMTP id 9F16EAC5B2 23, 34 -- for {Microscopy-at-microscopy.com} ; Fri, 25 Aug 2006 11:15:50 +0300 (IDT) 23, 34 -- Received: from td-mt-exchange.mt.technion.ac.il ([132.68.209.6]) by td-cc-mail.cc.technion.ac.il with Microsoft SMTPSVC(6.0.3790.1830); 23, 34 -- Fri, 25 Aug 2006 11:17:15 +0300 23, 34 -- Subject: Micro Saw 23, 34 -- Date: Fri, 25 Aug 2006 11:17:15 +0300 23, 34 -- Message-ID: {CBF88790B21AA6499C677A30D6E3D9E3135659-at-td-mt-exchange.mt.technion.ac.il} 23, 34 -- MIME-Version: 1.0 23, 34 -- X-MS-Has-Attach: 23, 34 -- Content-Type: text/plain; 23, 34 -- charset="utf-8" 23, 34 -- X-MS-TNEF-Correlator: 23, 34 -- Thread-Topic: Micro Saw 23, 34 -- content-class: urn:content-classes:message 23, 34 -- Thread-Index: AcbIJ0RfdPWO0EWJT42+tmtkolQjRw== 23, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6487.1 23, 34 -- From: {kaplan-at-mt.technion.ac.il} 23, 34 -- To: {Microscopy-at-microscopy.com} 23, 34 -- X-OriginalArrivalTime: 25 Aug 2006 08:17:15.0319 (UTC) FILETIME=[DFA2A470:01C6C81E] 23, 34 -- Content-Transfer-Encoding: 8bit 23, 34 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k7P8HGe4016836 ==============================End of - Headers==============================
Will anyone have a preparation protocol for 1% (TCH) Thiocarbohydrozide? I'm working with some fastidious mammal tissue for SEM and I was thinking on putting the tissue through OTO method, but I need a preparation protocol for 1%(TCH). I've been doing research on a preparation protocol with no success. I tried to mix the TCH in water to make a 1% mixture but I have difficulty getting the TCH to dissolve in DD water.
Thank You in advance
Omayra Velez MS Electron Microscopy/Histology Life Cell Corporation One Millennium Way Branchburg, NJ 08876 w. 908-947-1366 f. 908-947-1200
==============================Original Headers============================== 4, 18 -- From mayas003-at-yahoo.com Fri Aug 25 09:22:41 2006 4, 18 -- Received: from web84111.mail.mud.yahoo.com (web84111.mail.mud.yahoo.com [68.142.206.198]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7PEMfiI032531 4, 18 -- for {Microscopy-at-MSA.Microscopy.com} ; Fri, 25 Aug 2006 09:22:41 -0500 4, 18 -- Received: (qmail 51882 invoked by uid 60001); 25 Aug 2006 14:22:41 -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=NOqCY81NA3P4lusCUr4v4kZEN+iUfetyREUrGjnDm6b+HYMsMn1hx/LCfobdYC2OmnHinHvsQT8Qd5EPr++gB2R4GAvewgTJbu8Hl2fCp2jDaFTw7zPC4ICwb5VFi2yod4OuIRN7iG043b3/ZGrEx3ZjIOgtwryofdlz2yjz10w= ; 4, 18 -- Message-ID: {20060825142241.51880.qmail-at-web84111.mail.mud.yahoo.com} 4, 18 -- Received: from [63.81.204.49] by web84111.mail.mud.yahoo.com via HTTP; Fri, 25 Aug 2006 07:22:41 PDT 4, 18 -- Date: Fri, 25 Aug 2006 07:22:41 -0700 (PDT) 4, 18 -- From: Omayra Velez {mayas003-at-yahoo.com} 4, 18 -- Subject: TCH 4, 18 -- To: Microscopy-at-MSA.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==============================
I believe the original references suggested dissolving the TCH at 60 C for 1 hr with occasional vigorous agitation. I generally pre-heat some dH2O to 50-60 C and then add 1% TCH and sonicate for 15 min - it almost all goes into solution - just to be sure I let it sit for another 60 min in my 60 C oven. I generally filter with a 0.1 or 0.2 um filter before use. One other useful tip is after the first osmium, rinse well with dH2O and switch the tissue to a fresh vial. this will minimize any traces of osmium on the vial or cap that cause ugly large precipitates of osmium when you add the TCH. when we are being super careful, we switch after the second osmium step also. I like the technique and have had success with it. Good luck.
At 09:23 AM 08/25/06, 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
A search for "microscopy" in the Journal of Wood and Fiber Science http://www.swst.org/journal.html yielded a number of interesting hits. You can also check in the TAPPI Journal (www.tappi.org). If you can get the Nordic Pulp and Paper Research Journal, this is a good reference: "The ultrastructure of wood fibre surfaces as shown by a variety of microscopic methods - a review", I. Duchesne & G. Daniel, V 14, N 2, 1999. P. 129-139. Includes FE-SEM, TEM, and AFM. In my chapter "Electron Microscopy for the Pulp & Paper Industry" in "Industrial Applications of Electron Microscopy" (Li, 2003), I reproduced Gary Baum's drawing of the the macro to micro structures of wood fiber in paper. [Disclaimer: I may eventually get some royalties]
David Rothbard Bureau of Engraving and Printing
tina-at-pbrc.hawaii.edu wrote:
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==============================Original Headers============================== 7, 20 -- From rothbardd-at-netscape.net Fri Aug 25 11:45:07 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 k7PGj7e5032316 7, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 25 Aug 2006 11:45:07 -0500 7, 20 -- Received: from rothbardd-at-netscape.net 7, 20 -- by imo-d01.mx.aol.com (mail_out_v38_r7.6.) id w.4da.16d7fb6 (16239) 7, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 25 Aug 2006 12:45:03 -0400 (EDT) 7, 20 -- Received: from netscape.net (mow-d23.webmail.aol.com [205.188.139.164]) by air-in03.mx.aol.com (v112.5) with ESMTP id MAILININ33-3f6f44ef290e283; Fri, 25 Aug 2006 12:45:02 -0400 7, 20 -- Date: Fri, 25 Aug 2006 12:44:25 -0400 7, 20 -- From: rothbardd-at-netscape.net 7, 20 -- To: Microscopy-at-microscopy.com 7, 20 -- Subject: RE: [Microscopy] AFM/SEM of wood/plant material 7, 20 -- MIME-Version: 1.0 7, 20 -- Message-ID: {148E9208.44853A89.034D9A6A-at-netscape.net} 7, 20 -- X-Mailer: Atlas Mailer 2.0 7, 20 -- X-AOL-IP: 199.196.144.16 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://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] Nuclear Envelope EM protocol
Question: Hello ListServers:
Would anyone have a good TEM protocol for fixing/processing Xenopus nuclear envelope isolates? I would like to be able to image the double membranes and nuclear pores.
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Email: Phil.Ahrenkiel-at-sdsmt.edu Name: Phil Ahrenkiel
Organization: South Dakota School ofMines & Technology
Title-Subject: [Filtered] need STEM/turn of shutter
Question: Hi-
I ordered the KeenView with my startup before moving here to use with an Hitachi H-7000. We got a bottom-mount camera, because I heard there was no chance to do lattice imaging with a top-mount (don't know if that's true). The KeenView isn't retractable, and when the engineer got here, we realized that we had to remove the STEM detector to mount the CCD. The images are fine, but now I have no STEM detector, and I am wondering if anyone knows some other way to do STEM. In principle, we could use the KeenView to do STEM, and even do dark-field STEM, but I doubt it's that easy. We could also mount a STEM detector higher up, maybe in a 35-mm port. If I could find a top-mount CCD that could also do STEM, we would be in great shape.
We also have an SEM detector. Any ideas on how to take the images digitally on a PC? All we have is Polaroid. I see some available ports on the scanning unit, but I don't have any info on how to set up the interface.
The other thing is we have to trick the H-7000 not to take a film exposure when we lift the screen to use the CCD. Does anyone know a way to just deactivate the film camera (and shutter) temporarily? Thanks. ------------------- Phil Ahrenkiel, Assistant Professor Nanoscience and Nanoengineering Ph.D. Program South Dakota School of Mines and Technology 501 East St. Joseph Street Rapid City, South Dakota 57701 Phone: (605)394-5238, Fax: (605)394-5266 Email: Phil.Ahrenkiel-at-sdsmt.edu
perhaps you might have a pdf (if so, please allow for sending to your e.mail-address) of the following article:
Atherosclerosis 180 (2005) 293-303 Calcification of elastic fibers in human atherosclerotic plaque by Yuri V. Bobryshev Excerpt from Methods: 2.4.2.2. Identification of neutral lipids using osmium-thiocarbohydrazide-osmium technique. The osmium-thiocarbohydrazide-osmium (OTO) technique enables the visualization of neutral lipids in arterial tissue [7 References]. A number of previous studies that utilized OTO technique demonstrated that neutral lipids are associated with elastic fibers in atherosclerotic lesions [7 references]. In the present study, OTO technique described by Guyton et al. [ref] was carried out as previously [ref]. This involved an initial treatment of tissue samples with aqueous 1% OsO4, extensive washing in 0.1Mcacodylate buffer and then in distilled water, treatment with 1.0% aqueous thiocarbohydratezide (Sigma) for 5 min, washing with the buffer and water, and repeated fixation in aqueous 1% OsO4 for 10 min. The thiocarbohydrazyde solution was freshly prepared by intermittent shaking for 3-4h at 58 degr.C, cooling to 25 .C and filtration through a 0.22 ?m filter as used previously [Ref].
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Hello all:
Will anyone have a preparation protocol for 1% (TCH) Thiocarbohydrozide? I'm working with some fastidious mammal tissue for SEM and I was thinking on putting the tissue through OTO method, but I need a preparation protocol for 1%(TCH). I've been doing research on a preparation protocol with no success. I tried to mix the TCH in water to make a 1% mixture but I have difficulty getting the TCH to dissolve in DD water.
Thank You in advance
Omayra Velez MS Electron Microscopy/Histology Life Cell Corporation One Millennium Way Branchburg, NJ 08876 w. 908-947-1366 f. 908-947-1200
==============================Original Headers============================== 4, 18 -- From mayas003-at-yahoo.com Fri Aug 25 09:22:41 2006 4, 18 -- Received: from web84111.mail.mud.yahoo.com (web84111.mail.mud.yahoo.com [68.142.206.198]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7PEMfiI032531 4, 18 -- for {Microscopy-at-MSA.Microscopy.com} ; Fri, 25 Aug 2006 09:22:41 -0500 4, 18 -- Received: (qmail 51882 invoked by uid 60001); 25 Aug 2006 14:22:41 -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:C ontent-Transfer-Encoding; 4, 18 -- b=NOqCY81NA3P4lusCUr4v4kZEN+iUfetyREUrGjnDm6b+HYMsMn1hx/LCfobdYC2OmnHi nHvsQT8Qd5EPr++gB2R4GAvewgTJbu8Hl2fCp2jDaFTw7zPC4ICwb5VFi2yod4OuIRN7iG04 3b3/ZGrEx3ZjIOgtwryofdlz2yjz10w= ; 4, 18 -- Message-ID: {20060825142241.51880.qmail-at-web84111.mail.mud.yahoo.com} 4, 18 -- Received: from [63.81.204.49] by web84111.mail.mud.yahoo.com via HTTP; Fri, 25 Aug 2006 07:22:41 PDT 4, 18 -- Date: Fri, 25 Aug 2006 07:22:41 -0700 (PDT) 4, 18 -- From: Omayra Velez {mayas003-at-yahoo.com} 4, 18 -- Subject: TCH 4, 18 -- To: Microscopy-at-MSA.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============================== 14, 39 -- From W.Muss-at-salk.at Mon Aug 28 07:16:29 2006 14, 39 -- Received: from hermes.salk.at (hermes.lks.at [193.170.167.9]) 14, 39 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7SCGSMa019301 14, 39 -- for {Microscopy-at-MSA.Microscopy.com} ; Mon, 28 Aug 2006 07:16:29 -0500 14, 39 -- Received: from localhost (localhost [127.0.0.1]) 14, 39 -- by hermes.salk.at (Postfix) with ESMTP id AC0ACC386C; 14, 39 -- Mon, 28 Aug 2006 14:16:26 +0200 (CEST) 14, 39 -- X-Virii-Scanned: Kaspersky Antivirus at salk.at 14, 39 -- Received: from hermes.salk.at ([127.0.0.1]) 14, 39 -- by localhost (n1ex218.lks.local [127.0.0.1]) (amavisd-new, port 10024) 14, 39 -- with ESMTP id IE6qc2VqvYsW; Mon, 28 Aug 2006 14:16:26 +0200 (CEST) 14, 39 -- Received: from hermes.lks.at (hermes.salk.at [192.168.13.210]) 14, 39 -- by hermes.salk.at (Postfix) with ESMTP id 37022C384F; 14, 39 -- Mon, 28 Aug 2006 14:16:26 +0200 (CEST) 14, 39 -- Received: from localhost (localhost [127.0.0.1]) 14, 39 -- by hermes.lks.at (Postfix) with ESMTP id 154E35A9022; 14, 39 -- Mon, 28 Aug 2006 14:16:26 +0200 (CEST) 14, 39 -- Received: from hermes.lks.at ([127.0.0.1]) 14, 39 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 14, 39 -- with ESMTP id 81844-04; Mon, 28 Aug 2006 14:16:25 +0200 (CEST) 14, 39 -- Received: from c1pa003 (unknown [192.168.42.3]) 14, 39 -- by hermes.lks.at (Postfix) with SMTP id C39175A900D; 14, 39 -- Mon, 28 Aug 2006 14:16:25 +0200 (CEST) 14, 39 -- Received: by localhost with Microsoft MAPI; Mon, 28 Aug 2006 14:16:24 +0200 14, 39 -- Message-ID: {01C6CAAC.8B6251E0.W.Muss-at-salk.at} 14, 39 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 14, 39 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 14, 39 -- To: "'mayas003-at-yahoo.com'" {mayas003-at-yahoo.com} , 14, 39 -- "'Microscopy-at-MSA.Microscopy.com'" {Microscopy-at-MSA.Microscopy.com} 14, 39 -- Subject: [Microscopy] Re: TCH, thiocarbohydrazide prep 14, 39 -- Date: Mon, 28 Aug 2006 14:16:23 +0200 14, 39 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 14, 39 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 14, 39 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 14, 39 -- MIME-Version: 1.0 14, 39 -- Content-Type: text/plain; charset="us-ascii" 14, 39 -- Content-Transfer-Encoding: 7bit 14, 39 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at 14, 39 -- X-Scanned-By: SALK-Content-Filter ==============================End of - Headers==============================
Dear Phil, I have a Hitachi H-800, which is older and a 200 kV version of your H-7000. I have successfully integrated the Quartz PCI system (www.quartzimaging.com) to the STEM system to acquire images. If you still have the scan generator and control system active on the H-7000, you should be able to acquire SEM images and hook up an image acquire system. If you only disconnected the STEM detector, the scanning system should still work. This is activated by the STEM button on the controls. Normal TEM operation is the ZOOM button. Once you have pushed the STEM button, the right-hand lever will not activate the plate camera. I do not know where you can put your STEM detector, but the 35 mm port might work. A STEM detector is the same as a SEM detector, just below the sample, as far as I know. Best of 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: {Phil.Ahrenkiel-at-sdsmt.edu} To: {mager-at-interchange.ubc.ca} Sent: Sunday, August 27, 2006 8:49 AM
Hi everyone, Thanks for the excellent response. A colleague on campus was asking for the info and I have passed it on. At some point I am sure there will be others asking for this in our lab, so having something available will be an asset.
As you noticed, several people refered us to the Boulder campus site for IMOD which apparently has 3D and tomography capabilities.
Others provided names of less expensive (?) to expensive commercial softare packages. Let's face it, I've got champagne taste on a beer budget.
And of course, Image J was listed.
Thanks for your help - you're a great resource. John Shields EM Lab UGA Athens, GA John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602
706-542-4080
==============================Original Headers============================== 6, 21 -- From jpshield-at-uga.edu Mon Aug 28 16:49:24 2006 6, 21 -- Received: from puntd5.cc.uga.edu (puntd5.cc.uga.edu [128.192.1.108]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7SLnO44016630 6, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 28 Aug 2006 16:49:24 -0500 6, 21 -- Received: from punts4.cc.uga.edu (punts4.cc.uga.edu [128.192.1.115]) 6, 21 -- by puntd5.cc.uga.edu (MOS 3.7.4b-GA) 6, 21 -- with ESMTP id ACA61363; 6, 21 -- Mon, 28 Aug 2006 17:48:25 -0400 (EDT) 6, 21 -- Received: (from punts4.cc.uga.edu [128.192.63.198]) 6, 21 -- by punts4.cc.uga.edu (MOS 3.7.4c) 6, 21 -- with HTTPS/1.1 id BEJ46588 (AUTH jpshield-at-uga.edu); 6, 21 -- Mon, 28 Aug 2006 17:48:24 -0400 (EDT) 6, 21 -- From: John Shields {jpshield-at-uga.edu} 6, 21 -- Subject: results of TEM 3D 6, 21 -- To: msa listserve {Microscopy-at-MSA.Microscopy.Com} 6, 21 -- X-Mailer: Mirapoint Webmail Direct 3.7.4c 6, 21 -- MIME-Version: 1.0 6, 21 -- Content-Type: text/plain; charset=us-ascii 6, 21 -- Content-Transfer-Encoding: 7bit 6, 21 -- Message-Id: {20060828174824.BEJ46588-at-punts4.cc.uga.edu} 6, 21 -- Date: Mon, 28 Aug 2006 17:48:24 -0400 (EDT) ==============================End of - Headers==============================
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Email: dyel-at-mail.nih.gov Name: Chip Dye
Organization: NIH-NICHD
Title-Subject: [Filtered] Mixing Glut. and OsO4
Question: Dear ListServers,
Are the issues if glutaraldehyde and osmium tetroxide are made up separately in sod cac. buffer then mixed together?
unfortunately I never tried such a combination GA-Os.....
cf. GEYER G. (ed, 1973) Ultrahistochemie (Ultrahistochemistry): .....} use of a mixture of aldehyde and osmium not recommendable since because of temperature-/time-dependent reactions of both fixatives the properties of that solution changes uncontrollable...Linking/protein precipitation always turned out to be less than with each of the fixative substances alone.....therefore it seems that there is no addition in fixing effects.... { (Ref.--} HOPWOOD D.[1970]: The reactions between formaldehyde, glutaraldehyde, and osmium tetroxide, and their fixation effects on bovine albumin and on tissue blocks. Histochemie 24, pp.50-64)
But: I am aware of several papers having dealt with such a combination of GA-OsO4 (part.reduced)-[primary] fixation. (cf: http://www.wormbook.org/chapters/www_intromethodscellbiology/intromethod scellbiology.html : Protocol 10: Osmium + Aldehyde one-step fixation for TEM (Dave Hall) )
Mixing result will depend on purity of ingredients, more or less rapid reduction of OsO4 also due to } impure { GA-solution occurs, perhaps also depending on the buffer solution you use.
As additional hints: cf: http://www.uga.edu/caur/temnote1.htm scroll down to Fixatives/Osmium Tetroxide/[OsO4] } .....Although osmium tetroxide is typically used as a secondary fixative it is often used in conjunction with glutaraldehyde in what is sometimes referred to as a glut/osmium cocktail. This mixture is particularly useful for fixing single cells. Because aldehydes can reduce osmium it is recommended that the fixation be carried out at 4 degree C for 30 min or less. The mixture should be made immediately prior to use. A similar chemical ruthenium tetroxide is sometimes used as a less expensive alternative to osmium tetroxide..... { and finally:
J Electron Microsc (Tokyo). 1978;27(2):149-51. A formaldehyde-osmium tetroxide mixture and a formaldehyde-glutaraldehyd e-osmium tetroxide mixture as fixative for electron microscopy. Takahashi M, Suzuki H, Yamamoto I. A pdf of that article I would be able to send to you on request
and, last but not least: Histochemie. 1969;19(2):162-4 Simultaneous glutaraldehyde-osmium tetroxide fixation with postosmication. An improved fixation procedure for electron microscopy of plant and animal cells. Franke WW, Krien S, Brown RM Jr. Summary A fixation procedure for electron microscopy is described which includes a simultaneous glutaraldehyde-OsO4 fixation followed by postosmication. This procedure was found to have considerable advantages in preserving structures of plant and animal cells.
Best wishes and good luck,
Wolfgang Muss Salzburg/Austria
---------- Von: dyel-at-mail.nih.gov[SMTP:dyel-at-mail.nih.gov] Antwort an: dyel-at-mail.nih.gov Gesendet: Dienstag, 29. August 2006 01:23 An: W.Muss-at-salk.at Betreff: [Microscopy] Mixing Glut. and OsO4
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Question: Dear ListServers,
Are the issues if glutaraldehyde and osmium tetroxide are made up separately in sod. cac. buffer then mixed together?
Colleagues, To add to the references thoughtfully provided by Wolfgang Muss, here is one more. This one does not mention the use of glut and osmium in the primary fix but indeed it does. Hope its helpful, Tobias
AU Follet-Gueye, ML Pagny, S Faye, L Gomord, V Driouich, A TI An improved chemical fixation method suitable for immunogold localization of green fluorescent protein in the Golgi apparatus of tobacco bright yellow (BY-2) cells SO JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 2003, 51 (7): 931 - 940 AB In plant systems, the green fluorescent protein (GFP) is increasingly used as a marker to study dynamics of the secretory apparatus using fluorescence microscopy. The purpose of this study was to immunogold localize the GFP, at the electron microscopic level, in a line of tobacco BY-2-cultured cells, expressing a GFP-tagged Golgi glycosyltransferase. To this end we have developed a simple, one-step chemical fixation method that allow good structural preservation and specific labeling with anti-GFP antibodies. Using this method, we have been able to show that an N-glycan GFP-tagged xylosyltransferase is specifically associated with Golgi stacks of BY-2 transformed cells and is preferentially located in medial cisternae. As an alternative to cryofixation methods, such as high-pressure freezing, which requires specialized and expensive equipment not available in most laboratories, this method offers researchers the opportunity to investigate GFP-tagged proteins of the endomembrane system in tobacco BY-2 cells. P
} } } Good morning } dear Chip Dye, } } unfortunately I never tried such a combination GA-Os..... } } cf. GEYER G. (ed, 1973) Ultrahistochemie (Ultrahistochemistry): } .....} use of a mixture of aldehyde and osmium not recommendable since } because of temperature-/time-dependent reactions of both fixatives the } properties of that solution changes uncontrollable...Linking/protein } precipitation always turned out to be less than with each of the fixative } substances alone.....therefore it seems that there is no addition in fixing } effects.... { } (Ref.--} HOPWOOD D.[1970]: The reactions between formaldehyde, } glutaraldehyde, and osmium tetroxide, and their fixation effects on bovine } albumin and on tissue blocks. Histochemie 24, pp.50-64) } } But: I am aware of several papers having dealt with such a combination of } GA-OsO4 (part.reduced)-[primary] fixation. } (cf: } http://www.wormbook.org/chapters/www_intromethodscellbiology/intromethod } scellbiology.html : } Protocol 10: Osmium + Aldehyde one-step fixation for TEM (Dave Hall) ) } } Mixing result will depend on purity of ingredients, more or less rapid } reduction of OsO4 also due to } impure { GA-solution occurs, perhaps also } depending on the buffer solution you use. } } As additional hints: } cf: http://www.uga.edu/caur/temnote1.htm } scroll down to Fixatives/Osmium Tetroxide/[OsO4] } } .....Although osmium tetroxide is typically used as a secondary fixative } it is often used in conjunction with glutaraldehyde in what is sometimes } referred to as a glut/osmium cocktail. This mixture is particularly useful } for fixing single cells. Because aldehydes can reduce osmium it is } recommended that the fixation be carried out at 4 degree C for 30 min or } less. The mixture should be made immediately prior to use. A similar } chemical ruthenium tetroxide is sometimes used as a less expensive } alternative to osmium tetroxide..... { } and finally: } } J Electron Microsc (Tokyo). 1978;27(2):149-51. } A formaldehyde-osmium tetroxide mixture and a formaldehyde-glutaraldehyd } e-osmium tetroxide mixture as fixative for electron microscopy. } Takahashi M, Suzuki H, Yamamoto I. } A pdf of that article I would be able to send to you on request } } } and, last but not least: } Histochemie. 1969;19(2):162-4 } Simultaneous glutaraldehyde-osmium tetroxide fixation with postosmication. } An improved fixation procedure for electron microscopy of plant and animal } cells. } Franke WW, Krien S, Brown RM Jr. } Summary A fixation procedure for electron microscopy is described which } includes a simultaneous glutaraldehyde-OsO4 fixation followed by } postosmication. This procedure was found to have considerable advantages in } preserving structures of plant and animal cells. } } Best wishes and good luck, } } Wolfgang Muss } Salzburg/Austria } } } } } ---------- } Von: dyel-at-mail.nih.gov[SMTP:dyel-at-mail.nih.gov] } Antwort an: dyel-at-mail.nih.gov } Gesendet: Dienstag, 29. August 2006 01:23 } An: W.Muss-at-salk.at } Betreff: [Microscopy] Mixing Glut. and OsO4 } } ------------------------------------------------------------------------ } ---- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
SEM Technician position to be immediately filled at the Catholic University of America in Washington D.C.
Scanning Electron Microscopy Technician
Scanning electron microscopy (SEM) operator needed for routine analytical work associated with waste vitrification industry. The successful candidate must have a minimum of 200 hours operational experience with a full understanding of how instrument parameters affect microstructural characterization. Additionally, candidate must have a thorough understanding of energy dispersive spectroscopy (EDS,) particularly, as it relates to conducting the analysis.
This position requires an AAS in Electron Microscopy or a 4 year science based degree with 2-5 years hands on electron microscopy experience. SEM and EDS experience must be current.
• Hands on SEM experience with fundamental understanding of how variables affect microanalysis. • Hands on EDS experience with fundamental understanding of how SEM variables affect microanalysis, as well as, knowledge of pitfalls associated with this type of analysis. • Preparation of specimens for conducting SEM/EDS analysis. • Must have excellent English speaking/comprehension skills. • Must have excellent computer skills. • Must be a US citizen or have a valid work permit, (no sponsorship available.) • Hands on TEM experience and sample preparation a major plus. • Knowledge of minerals (mineral formation) also a bonus.
This is an entry level to intermediate level position with a salary range of 42 to 48K. Benefits include, (but not limited to,) 21 days paid vacation, (plus a 2 week Christmas break,) 2x matching 401k, (after 1 year of employment,) and tuition reimbursement, (2 classes per semester.) Prospective employee(s) will be asked to submit several examples of their analytical work or show a portfolio during their interview, as well as, demonstrate on the instrumentation their competency.
Send resumes and cover letters to Andrew Buechele, PhD. at andrewb-at-vsl.cua.edu – CC to Cavin Mooers at cavinm-at-vsl.cua.edu Phone interviews will be conducted over the course of the next month, with onsite visits thereafter.
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This Question was submitted to Ask-A-Microscopist by (mfein-at-bu.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, August 29, 2006 at 09:59:40 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both mfein-at-bu.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: mfein-at-bu.edu Name: Marcia D. Feinberg
Organization: Boston University
Education: Graduate College
Location: Boston, MA
Title: seeking parts to buy for a Leitz Ortholux 1 light microscope:
Question: Hello! We are looking for 1) a light socket assembly and 2) a stage slide holder for our original, black body style, Lietz Ortholux 1, light microscope. Please contact Marcia at mfein-at-bu.edu, Thank you!
Beth, Not sure why anyone would prefer JB4 to my nice mix of butylmethylmetacrylate (BMM). Maybe there are reasons. If you want a protocol for BMM, please contact me off line.
Tobias
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Email: klangwor-at-uoregon.edu Name: Kurt Langworthy
Organization: University of Oregon
Title-Subject: [Filtered] E-beam lithography
Question: Does anybody have insight on how to create 3-D structures of varying height using e-beam lithography, or is a dual beam FIB required for such tasks? The project presented to me this morning would require making small columns separated by varying intervals with controlled vertical heights up to 13nm. Best Regards, Kurt Langworthy
Hello Everyone, Recently I been in a discussion about how many pixels a feature should contain to provide meaningful results from image analysis. For example, if I threshold an image or measure a perimeter how many pixels do I need a feature to have as to insure I have "statistical meaningful" data. It seem intuitive that I should have as many as possible, but what about a particle or feature that has only 12 pixels maximum in one direction (say a fiber)?
I realize I could have a rectangle 9 by 7 pixels which would give me a diagonal of 11 pixels, but if I could only measure features that had at least 10 pixels would this feature have meaning?
I think I've expressed myself as clearly as my current understand is will allow me, I'll turn this question loose on the high tension thinkers out there!
Thanks
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, 17 -- From frank.karl-at-degussa.com Wed Aug 30 10:24:53 2006 10, 17 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 10, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7UFOp01018353 10, 17 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 10:24:52 -0500 10, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 10, 17 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k7UFOhD0022271 10, 17 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 17:24:49 +0200 10, 17 -- Subject: Question: pixels, pixels, little pixels.... 10, 17 -- To: microscopy-at-msa.microscopy.com 10, 17 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 10, 17 -- Message-ID: {OF10988FCD.61D70032-ON852571DA.0052EAEE-852571DA.0054A7DA-at-degussa.com} 10, 17 -- From: frank.karl-at-degussa.com 10, 17 -- Date: Wed, 30 Aug 2006 11:24:39 -0400 10, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 10, 17 -- 08/30/2006 10:24:49 AM 10, 17 -- MIME-Version: 1.0 10, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
} Recently I been in a discussion about how many pixels a } feature should contain to provide meaningful results from } image analysis. For example, if I threshold an image or } measure a perimeter how many pixels do I need a feature to } have as to insure I have "statistical meaningful" data. It } seem intuitive that I should have as many as possible, but } what about a particle or feature that has only 12 pixels } maximum in one direction (say a fiber)? } } I realize I could have a rectangle 9 by 7 pixels which would } give me a diagonal of 11 pixels, but if I could only measure } features that had at least 10 pixels would this feature have meaning? } } I think I've expressed myself as clearly as my current } understand is will allow me, I'll turn this question loose on } the high tension thinkers out there!
I too have given this much thought, without finding any reference to any work rgarding an error analysis for pixel segmentation. To a 1st approximation, significance (or confidence) is probably tied to counting error. This would make a single pixel insignificant, and imply a 10% error for a feature with 100 pixels. However, I've often thought that 2 features, both with same number of pixels but both with different perimeters, must have different error ... Or, at least it cannot be as simple as counting error.
Error analysis is also complicated by the process by which you segment features. For example, I might segment for features with 2 algorithms, and both seemingly and visually provide good results. However, because of the precision associated with 8bits (as well as where in the histogram that value might be), the 2 resulting values can differ as much as 10%, and have nothing to do with "number of pixels"!
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 8, 21 -- From Michael-at-Shaffer.net Wed Aug 30 11:19:41 2006 8, 21 -- Received: from ws6-8.us4.outblaze.com (ws6-8.us4.outblaze.com [205.158.62.24]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7UGJeJv029176 8, 21 -- for {Microscopy-at-microscopy.com} ; Wed, 30 Aug 2006 11:19:40 -0500 8, 21 -- Received: (qmail 29916 invoked from network); 30 Aug 2006 16:20:10 -0000 8, 21 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) 8, 21 -- by ws6-8.us4.outblaze.com with SMTP; 30 Aug 2006 16:20:08 -0000 8, 21 -- From: "michael shaffer" {michael-at-Shaffer.net} 8, 21 -- To: {frank.karl-at-degussa.com} 8, 21 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 8, 21 -- Subject: RE: [Microscopy] Question: pixels, pixels, little pixels.... 8, 21 -- Date: Wed, 30 Aug 2006 13:49:29 -0230 8, 21 -- Message-ID: {002a01c6cc50$133a5dc0$8d829986-at-roamingwolf} 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-Mailer: Microsoft Office Outlook 11 8, 21 -- Thread-Index: AcbMSLJviMzzPtqVQre7k+t3KhWvCgABRZsg 8, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 8, 21 -- In-Reply-To: {200608301525.k7UFPZDC019104-at-ns.microscopy.com} ==============================End of - Headers==============================
Nyquist's Theorem is the general sampling theorem used in microscopy. While the exact interpretation is that you need 2.3 pixels/object (assumedly, per dimension, if you are doing sizing), most of us use a practical limit somewhere between 3 and 10.
Hope that 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 December. Call us today for details.
P. S. Need a good general reference or light microscopy text for the Fall 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 11:32 AM 8/30/2006, michael-at-Shaffer.net wrote:
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==============================Original Headers============================== 13, 18 -- From bfoster-at-mme1.com Wed Aug 30 12:39:16 2006 13, 18 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 13, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7UHdGGn018299 13, 18 -- for {microscopy-at-microscopy.com} ; Wed, 30 Aug 2006 12:39:16 -0500 13, 18 -- Received: (qmail 20626 invoked by uid 2020); 30 Aug 2006 12:58:17 -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 (DHE-RSA-AES256-SHA encrypted) SMTP; 30 Aug 2006 12:58:17 -0500 13, 18 -- Message-Id: {7.0.1.0.0.20060830120702.0203b120-at-mme1.com} 13, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 18 -- Date: Wed, 30 Aug 2006 12:39:26 -0500 13, 18 -- To: michael-at-Shaffer.net, microscopy-at-microscopy.com 13, 18 -- From: Barbara Foster {bfoster-at-mme1.com} 13, 18 -- Subject: Re: [Microscopy] RE: Question: pixels, pixels, little 13, 18 -- pixels.... 13, 18 -- In-Reply-To: {200608301622.k7UGMGa4000766-at-ns.microscopy.com} 13, 18 -- References: {200608301622.k7UGMGa4000766-at-ns.microscopy.com} 13, 18 -- Mime-Version: 1.0 13, 18 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
On Aug 30, 2006, at 9:19 AM, michael-at-Shaffer.net wrote:
} Frank writes ... } } } Recently I been in a discussion about how many pixels a } } feature should contain to provide meaningful results from } } image analysis. For example, if I threshold an image or } } measure a perimeter how many pixels do I need a feature to } } have as to insure I have "statistical meaningful" data. It } } seem intuitive that I should have as many as possible, but } } what about a particle or feature that has only 12 pixels } } maximum in one direction (say a fiber)? } } } } I realize I could have a rectangle 9 by 7 pixels which would } } give me a diagonal of 11 pixels, but if I could only measure } } features that had at least 10 pixels would this feature have meaning? } } } } I think I've expressed myself as clearly as my current } } understand is will allow me, I'll turn this question loose on } } the high tension thinkers out there! } } I too have given this much thought, without finding any reference to } any } work rgarding an error analysis for pixel segmentation. To a 1st } approximation, significance (or confidence) is probably tied to } counting } error. This would make a single pixel insignificant, and imply a 10% } error } for a feature with 100 pixels. However, I've often thought that 2 } features, } both with same number of pixels but both with different perimeters, } must } have different error ... Or, at least it cannot be as simple as } counting } error. } } Error analysis is also complicated by the process by which you segment } features. For example, I might segment for features with 2 } algorithms, and } both seemingly and visually provide good results. However, because of } the } precision associated with 8bits (as well as where in the histogram that } value might be), the 2 resulting values can differ as much as 10%, and } have } nothing to do with "number of pixels"! } Dear Frank & Michael, The highest spatial frequency at which one can hope to collect information, called the Nyquist frequency, is such that the wavelength is 2 pixels. In practice one would have difficulty collecting information from greater than 2/3 Nyquist, corresponding to a feature comparable to 3 pixels. Of course, such considerations as noise and contrast limit the significance of the information collected, but if one can combine images of identical particles, these limitations can be overcome to an extent, and one can demonstrate that the average images do, indeed, contain information at the 2 to 3 pixel level. Microscope performance, as measured by the contrast transfer function, will also limit the information available, but for larger objects at lower magnifications the CTF should be nearly 1. Thus there are different considerations for cryoEM of radio-labile, low-contrast specimens vs radiation-resistant, higher-Z specimens. For specimens that are thin in one direction, such as the fiber you mentioned, the point-spread function of the detector must also be considered (this is also true for specimens that are small in both directions). The PSF will smear out an edge over several pixels, and it will also lower the contrast of your feature, since the same signal will now be spread over a larger range. A fiber that has the same width as the PSF and a signal that is twice the background noise will now appear to be anywhere from much smaller than the PSF to about twice as wide as the PSF with a signal about 50% larger than the background. In this case, measuring the diameter at many points along its length and deconvoluting the PSF could allow one to recover the width accurately. The bottom line is that ' to insure I have "statistical meaningful" data' is a complicated issue. 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 Aug 30 12:46:44 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 k7UHkgni028584 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 12:46:43 -0500 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 1F02D10A2B8 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 10:46:42 -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 5D13B2F064 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 10:46:40 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200608301619.k7UGJlBj029340-at-ns.microscopy.com} 4, 22 -- References: {200608301619.k7UGJlBj029340-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {8545c983ac9c18be03a746a9bf6e5334-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] RE: Question: pixels, pixels, little pixels.... 4, 22 -- Date: Wed, 30 Aug 2006 10:49:32 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
What would the material be that is 13nm thick/high? What thickness is when it is the thinest?
What are the dimensional spacings between each column?
Depending on details, one might lay down the thin material and use conventional photolithography to pattern the wafer or substrate. Another key dimension is the diameter of the columns.
E-beam litho would have a tough time if the pitch is too small. So would a laser writer. Depending on pitch and minimum feature size, I would suspect that standard litho would work or at worst, one or two phase masks.
gary g.
At 04:12 PM 8/29/2006, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Wed Aug 30 14:06:41 2006 11, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k7UJ6eel007931 11, 20 -- for {microscopy-at-microscopy.com} ; Wed, 30 Aug 2006 14:06:40 -0500 11, 20 -- Received: (qmail 31053 invoked from network); 30 Aug 2006 12:06:40 -0700 11, 20 -- Received: by simscan 1.1.0 ppid: 31046, pid: 31048, t: 0.1655s 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 qsmtp3 with SMTP; 30 Aug 2006 12:06:40 -0700 11, 20 -- Message-Id: {7.0.1.0.2.20060830120111.0253cb78-at-gaugler.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 20 -- Date: Wed, 30 Aug 2006 12:06:40 -0700 11, 20 -- To: klangwor-at-uoregon.edu 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] viaWWW: E-beam lithography 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200608292312.k7TNCjNw015606-at-ns.microscopy.com} 11, 20 -- References: {200608292312.k7TNCjNw015606-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 want to ignore the question of statistics since you seem to have a SNR high enough to do thresholding and segmentation.
The sampling theorem is only valid if the scene you digitize contains no spatial frequencies beyond some maximum value. In that case the digital image is completely equivalent to the original scene (in the sense that you can recreate the original scene with arbitrary sampling) if the highest frequency wave is represented by at least two pixels per wavelength. I. e. the Nyquist frequency is at least as high as the highest frequency contained in the scene. In this case there is no point in reducing the pixel size.
The situation is different if a scene contains higher frequencies, no matter whether these are noise or signal (assuming you have some way of telling the difference). Frequencies in the scene lying above Nyquist will appear in the digital image as aliases, that is frequencies below Nyquist that don't actually exist in the scene. I believe one reason why 1.5-fold oversampling is often recommended is to reduce the effects of noise-aliasing (noise decreases with frequency). If the scene contains higher frequencies other than noise the digitized image will show features that are not present in the scene and any segmentation will produce meaningless results. For example try taking a digital photograph of a fence from a distance such that the pixelsize becomes larger than the width of one board in the fence. In your picture you'll see something resembling a fence, but the spacings will be completely wrong (much too big).
In summary: I don't think more pixels is better, but it might be worse. Choose the pixel size based on the contents of the scene you want to digitize.
If somebody can explain the merits of oversampling 1.5-fold to me (beyond the simple explanation I've offered) I would be grateful.
Philip
Philip Koeck Karolinska Institute and Södertörns Högskola Dept. of Bioscience at Novum tel: +46 8 608 9186 fax: +46 8 608 9290 web: http://www.csb.ki.se/users/philip/philown.html
-----Original Message----- X-from: frank.karl-at-degussa.com [mailto:frank.karl-at-degussa.com] Sent: 30 August 2006 17:29 To: Philip.Koeck-at-biosci.ki.se
Hello Everyone, Recently I been in a discussion about how many pixels a feature should contain to provide meaningful results from image analysis. For example, if I threshold an image or measure a perimeter how many pixels do I need a feature to have as to insure I have "statistical meaningful" data. It seem intuitive that I should have as many as possible, but what about a particle or feature that has only 12 pixels maximum in one direction (say a fiber)?
I realize I could have a rectangle 9 by 7 pixels which would give me a diagonal of 11 pixels, but if I could only measure features that had at least 10 pixels would this feature have meaning?
I think I've expressed myself as clearly as my current understand is will allow me, I'll turn this question loose on the high tension thinkers out there!
Thanks
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
==============================Original Headers============================== 22, 27 -- From Philip.Koeck-at-biosci.ki.se Thu Aug 31 04:52:36 2006 22, 27 -- Received: from smtp.biosci.ki.se (smtp.biosci.ki.se [130.237.109.196]) 22, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7V9qZTe000503 22, 27 -- for {microscopy-at-msa.microscopy.com} ; Thu, 31 Aug 2006 04:52:35 -0500 22, 27 -- Received: from smtp.biosci.ki.se (localhost [127.0.0.1]) 22, 27 -- by localhost (Postfix) with ESMTP id 107D54FF01; 22, 27 -- Thu, 31 Aug 2006 11:52:34 +0200 (CEST) 22, 27 -- Received: from whale.csb.ki.se (whale.csb.ki.se [130.237.109.8]) 22, 27 -- by smtp.biosci.ki.se (Postfix) with SMTP id E722C4FEF1; 22, 27 -- Thu, 31 Aug 2006 11:52:33 +0200 (CEST) 22, 27 -- Received: from rapana.csb.ki.se by whale.csb.ki.se (5.65v4.0/1.1.10.5/03Feb98-0237PM) 22, 27 -- id AA06864; Thu, 31 Aug 2006 11:52:32 +0200 22, 27 -- Message-Id: {10608310952.AA06864-at-whale.csb.ki.se} 22, 27 -- From: "Philip Koeck" {Philip.Koeck-at-biosci.ki.se} 22, 27 -- To: {frank.karl-at-degussa.com} , {tivol-at-caltech.edu} , {bfoster-at-mme1.com} , 22, 27 -- {michael-at-Shaffer.net} , {microscopy-at-msa.microscopy.com} 22, 27 -- Subject: RE: [Microscopy] Question: pixels, pixels, little pixels.... 22, 27 -- Date: Thu, 31 Aug 2006 11:52:30 +0200 22, 27 -- Mime-Version: 1.0 22, 27 -- Content-Type: text/plain; 22, 27 -- charset="iso-8859-1" 22, 27 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 22, 27 -- In-Reply-To: {200608301528.k7UFSurT023224-at-ns.microscopy.com} 22, 27 -- X-Mimeole: Produced By Microsoft MimeOLE V6.00.2900.2962 22, 27 -- Thread-Index: AcbMSQLmBqclxWvSQuqNyg5HbF3FfAAkwW2Q 22, 27 -- Content-Transfer-Encoding: 8bit 22, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7V9qZTe000503 ==============================End of - Headers==============================
Two replies to different aspects of this question:
(1) Well, the reason for oversampling above Nyquist is actually fairly simple.
Think of the spatial frequencies (edges) as sine waves. Sampling at the Nyquist level of 2x the highest frequency allows you to sample the highest frequency incoming sine wave at peak and trough. Anything less and you will instead get a beat frequency from your samples. This is the _minimum_ sampling to get that high frequency.
Here's where I start reaching for the white-board pens...
However, if the high frequency is _out of phase_ with your samples, you might be perfectly sampling exactly where the input is crossing zero, resulting in no output data. Sampling right in phase gives maximum response, 90 degrees out of phase gives zero response. Hence the idea of sampling a little faster, as in 1.5x Nyquist, so that you get some peaks, some troughs, and can estimate the high frequency better.
(2) How many pixels? Depends on what you're measuring. You've got intensity variations, camera variations, illumination artifacts, segmentation bias, on and on and on. I would suggest taking your individual measurement(s) and looking at what happens statistically when you add/subtract some pixels. Diameter, area, shape, and integrated optical density will have different responses, linear, a ^2 relationship, or something else, based on how it's calculated.
Keep in mind that you're often polishing a pig! I have talked to people complaining about their 7th-8th decimal place of measurements varying (due to round-off of single precision floating point values), when uncorrected distortions of their images make their data accurate to _maybe_ +/-15 percent if they're lucky.
Minor shameless self-promotion here - I did a poster on imaging artifacts in microarrays a few years ago, including in particular segmentation errors. Take a look; I don't promise rocket science, but some of it might be useful:
-- Kevin Ryan Media Cybernetics, Inc. www.mediacy.com
} -----Original Message----- } From: Philip.Koeck-at-biosci.ki.se [mailto:Philip.Koeck-at-biosci.ki.se] } Sent: Thursday, August 31, 2006 5:55 AM } To: kevin-at-mediacy.com } Subject: [Microscopy] RE: Question: pixels, pixels, little pixels.... } } ... } } In summary: I don't think more pixels is better, but it might be } worse. } Choose the pixel size based on the contents of the scene you want to } digitize. } } If somebody can explain the merits of oversampling 1.5-fold to me } (beyond the simple explanation I've offered) I would be grateful. } } Philip } } } Philip Koeck } Karolinska Institute and Södertörns Högskola Dept. of Bioscience at } Novum } tel: +46 8 608 9186 } fax: +46 8 608 9290 } web: http://www.csb.ki.se/users/philip/philown.html }
==============================Original Headers============================== 18, 22 -- From kevin-at-mediacy.com Thu Aug 31 09:02:35 2006 18, 22 -- Received: from outbound.exchangemessage.net (outbound.exchangemessage.net [128.241.127.68]) 18, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7VE2Y20014457 18, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 31 Aug 2006 09:02:34 -0500 18, 22 -- Received: from wtrscp26021.exmsg.net ([128.241.125.43]) by outbound.exchangemessage.net with Microsoft SMTPSVC(6.0.3790.211); 18, 22 -- Thu, 31 Aug 2006 10:02:30 -0400 18, 22 -- Received: from KevinXP ([208.51.131.98]) by wtrscp26021.exmsg.net with Microsoft SMTPSVC(6.0.3790.211); 18, 22 -- Thu, 31 Aug 2006 10:02:33 -0400 18, 22 -- From: "Kevin Ryan" {kevin-at-mediacy.com} 18, 22 -- To: {Microscopy-at-microscopy.com} 18, 22 -- Subject: RE: [Microscopy] RE: Question: pixels, pixels, little pixels.... 18, 22 -- Date: Thu, 31 Aug 2006 10:02:24 -0400 18, 22 -- Message-ID: {003e01c6cd06$1660d360$820a0a0a-at-Mediacy.local} 18, 22 -- MIME-Version: 1.0 18, 22 -- Content-Type: text/plain; 18, 22 -- charset="iso-8859-1" 18, 22 -- X-Mailer: Microsoft Office Outlook 11 18, 22 -- Thread-Index: AcbM44IoKyZE1fD5QX6VZSLh5WCODwAHmcJAAAEHqwA= 18, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 18, 22 -- X-OriginalArrivalTime: 31 Aug 2006 14:02:33.0979 (UTC) FILETIME=[1B65C4B0:01C6CD06] 18, 22 -- Content-Transfer-Encoding: 8bit 18, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k7VE2Y20014457 ==============================End of - Headers==============================
Has anyone noticed a qualitative or quantitative difference in biological imaging with Leica oil index of refraction 1.518 vs Cargill Labs type DF oil 1.515?
Thanks. ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
==============================Original Headers============================== 3, 28 -- From cammer-at-aecom.yu.edu Thu Aug 31 11:18:00 2006 3, 28 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 3, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7VGI0ms027123 3, 28 -- for {microscopy-at-microscopy.com} ; Thu, 31 Aug 2006 11:18:00 -0500 3, 28 -- Received: from draco.aecom.yu.edu (draco.aecom.yu.edu [129.98.1.160]) 3, 28 -- by mailgw.aecom.yu.edu (8.12.11.20060308/8.12.11) with ESMTP id k7VGI0E1012047 3, 28 -- for {microscopy-at-microscopy.com} ; Thu, 31 Aug 2006 12:18:00 -0400 3, 28 -- Received: from draco.aecom.yu.edu (unknown [127.0.0.1]) 3, 28 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id 57C86329AD6 3, 28 -- for {microscopy-at-microscopy.com} ; Thu, 31 Aug 2006 12:17:03 -0400 (EDT) 3, 28 -- X-AuditID: 816201a0-aee42bb000007ca7-ef-44f70b7f174d 3, 28 -- Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.100]) 3, 28 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id 236D82E5725 3, 28 -- for {microscopy-at-microscopy.com} ; Thu, 31 Aug 2006 12:17:03 -0400 (EDT) 3, 28 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.30.137]) 3, 28 -- by post.aecom.yu.edu (Postfix) with ESMTP id C99AF28 3, 28 -- for {microscopy-at-microscopy.com} ; Thu, 31 Aug 2006 12:17:59 -0400 (EDT) 3, 28 -- Message-Id: {5.2.1.1.2.20060831121324.0115ca40-at-mailserver.aecom.yu.edu} 3, 28 -- X-Sender: cammer-at-mailserver.aecom.yu.edu 3, 28 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 3, 28 -- Date: Thu, 31 Aug 2006 12:15:58 -0400 3, 28 -- To: microscopy-at-microscopy.com 3, 28 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 3, 28 -- Subject: XYZ Z axis resolution 405nm to 700 nm imaging Leica 1.518 vs 3, 28 -- Cargill Labs DF oil 1.515 3, 28 -- Mime-Version: 1.0 3, 28 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 3, 28 -- X-Brightmail-Tracker: AAAAAA== ==============================End of - Headers==============================
We received an old Noran Vantage EDS system to use with our JEOL-2010 TEM. Unfortunately we don't have the original manuals, except for the material about the software. While I was connecting the cables, I did notice that some part of the equipment may be missing. What we have is:
1) Noran detector (already mounted in the column), 2) Detector power supply, 3) Detector position controller, 4) Noran Vantage Digital Acquisition System (DAS), 5) PC running Windows-NT with all the software.
The cable from the detector labeled "analyzer" with a DB15-male connector cannot be plugged anywhere, except on the back of the detector position controller (it doesn't make sense to me). The DAS do have only one external SCSI connector on the back, nothing else. Can you confirm if we are missing any electronic box from the list above? Probably some kind of signal processor? I will appreciate any information.
Regards,
Carlos
o-------------------------------------------------------o | Carlos Kazuo Inoki | | Department of Physics - University at Albany | | 1400 Washington Ave.- Albany - NY - 12222 | o-------------------------------------------------------o
==============================Original Headers============================== 7, 20 -- From kazuo-at-csc.albany.edu Thu Aug 31 15:24:01 2006 7, 20 -- Received: from smtp.albany.edu (mail1.its.albany.edu [169.226.1.105]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7VKO0EX010021 7, 20 -- for {Microscopy-at-Microscopy.Com} ; Thu, 31 Aug 2006 15:24:01 -0500 7, 20 -- Received: from unix1.its.albany.edu (unix2.its.albany.edu [169.226.1.39]) 7, 20 -- by smtp.albany.edu (8.13.6/8.13.6) with ESMTP id k7VKNv4W001035 7, 20 -- for {Microscopy-at-Microscopy.Com} ; Thu, 31 Aug 2006 16:23:59 -0400 (EDT) 7, 20 -- Received: from localhost (kazuo-at-localhost) 7, 20 -- by unix1.its.albany.edu (8.13.6/8.13.6/Submit) with ESMTP id k7VKNqst004419 7, 20 -- for {Microscopy-at-Microscopy.Com} ; Thu, 31 Aug 2006 16:23:57 -0400 (EDT) 7, 20 -- X-Authentication-Warning: unix1.its.albany.edu: kazuo owned process doing -bs 7, 20 -- Date: Thu, 31 Aug 2006 16:23:52 -0400 (EDT) 7, 20 -- From: Carlos Kazuo Inoki {kazuo-at-csc.albany.edu} 7, 20 -- X-X-Sender: kazuo-at-unix1.its.albany.edu 7, 20 -- To: MSA Listserve {Microscopy-at-Microscopy.Com} 7, 20 -- Subject: Help with Noran EDS system 7, 20 -- Message-ID: {Pine.GSO.4.58.0608311603090.3848-at-unix1.its.albany.edu} 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: TEXT/PLAIN; charset=US-ASCII 7, 20 -- X-Scanned-By: MIMEDefang 2.57 on 169.226.1.44 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mbush-at-fit.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, August 31, 2006 at 11:12:00 ---------------------------------------------------------------------------
Email: mbush-at-fit.edu Name: Mark Bush
Organization: Florida Tech
Education: Graduate College
Location: Melbourne, Fl
Question: I have a 2nd floor lab that shares a common wall with the Air-conditioning plant for a large building. Consequently we get substantial vibration through wall and floor. I have regular lab benches and cannot replace them as I need the cupboard storage. On those benches I have six Zeiss Axioskop or Axiolab microscopes. We have tried dampening vibrations with foam and granite slabs, but that isn't very effective. We can't go to microscope tables, due to lack of space. Does anyone have experience with isolation devices, e.g. Vistek or similar? Do these work well? I wish I could say money wasn't an issue, but it is and splurging $9K on 6 Vistek would strain the budget. All solutions welcome!
I'm having Bill Miller send you his famous tennis ball suspension system. It is inexpensive and may help.
If that doesn't, contact Ken Piel at Nanotech-America (www.nanotech-america.com). They sell a variety of anti-vibration tables for their AFMs and may have one in your price range.
Caveat: MME works closely with NTA.
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 December. Call us today for details.
P. S. Need a good general reference or light microscopy text for the Fall 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 05:57 PM 8/31/2006, mbush-at-fit.edu wrote:
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==============================Original Headers============================== 19, 17 -- From bfoster-at-mme1.com Thu Aug 31 18:34:08 2006 19, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 19, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7VNY8h7001292 19, 17 -- for {microscopy-at-microscopy.com} ; Thu, 31 Aug 2006 18:34:08 -0500 19, 17 -- Received: (qmail 14565 invoked by uid 2020); 31 Aug 2006 18:53:17 -0500 19, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 19, 17 -- by enterprise.5starpro.com with (DHE-RSA-AES256-SHA encrypted) SMTP; 31 Aug 2006 18:53:16 -0500 19, 17 -- Message-Id: {7.0.1.0.0.20060831183145.020315a8-at-mme1.com} 19, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 19, 17 -- Date: Thu, 31 Aug 2006 18:34:16 -0500 19, 17 -- To: mbush-at-fit.edu, microscopy-at-microscopy.com 19, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 19, 17 -- Subject: Re: [Microscopy] AskAMicroscopist: Vibration Solutions Needed 19, 17 -- In-Reply-To: {200608312250.k7VMo6xl026774-at-ns.microscopy.com} 19, 17 -- References: {200608312250.k7VMo6xl026774-at-ns.microscopy.com} 19, 17 -- Mime-Version: 1.0 19, 17 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
I respectfully refuse to buy what you say in point 1.
I agree that a wave at Nyquist frequency, which is aligned with the rows of pixels will in general be downweighted (and in the worst case deleted completely) due to the phase-mismatch you describe. However, this applies to exactly two plane waves in the entire spectrum, assuming that the scene to be digitized has a maximum frequency equal to Nyquist. Waves that run at an angle to the rows of pixels will be perfectly reconstructable (given all the relevant transfer functions). This is a minor problem, in my opinion, and doesn't require 1.5-fold oversampling. (1.1-fold would be plenty if you insist on fixing it.)
Any other ideas?
Philip
-----Original Message----- X-from: kevin-at-mediacy.com [mailto:kevin-at-mediacy.com] Sent: 31 August 2006 16:06 To: Philip.Koeck-at-biosci.ki.se
Two replies to different aspects of this question:
(1) Well, the reason for oversampling above Nyquist is actually fairly simple.
Think of the spatial frequencies (edges) as sine waves. Sampling at the Nyquist level of 2x the highest frequency allows you to sample the highest frequency incoming sine wave at peak and trough. Anything less and you will instead get a beat frequency from your samples. This is the _minimum_ sampling to get that high frequency.
Here's where I start reaching for the white-board pens...
However, if the high frequency is _out of phase_ with your samples, you might be perfectly sampling exactly where the input is crossing zero, resulting in no output data. Sampling right in phase gives maximum response, 90 degrees out of phase gives zero response. Hence the idea of sampling a little faster, as in 1.5x Nyquist, so that you get some peaks, some troughs, and can estimate the high frequency better.
-- Kevin Ryan Media Cybernetics, Inc. www.mediacy.com
} -----Original Message----- } From: Philip.Koeck-at-biosci.ki.se [mailto:Philip.Koeck-at-biosci.ki.se] } Sent: Thursday, August 31, 2006 5:55 AM } To: kevin-at-mediacy.com } Subject: [Microscopy] RE: Question: pixels, pixels, little pixels.... } } ... } } In summary: I don't think more pixels is better, but it might be } worse. } Choose the pixel size based on the contents of the scene you want to } digitize. } } If somebody can explain the merits of oversampling 1.5-fold to me } (beyond the simple explanation I've offered) I would be grateful. } } Philip }
==============================Original Headers============================== 24, 25 -- From Philip.Koeck-at-biosci.ki.se Fri Sep 1 05:22:40 2006 24, 25 -- Received: from smtp.biosci.ki.se (smtp.biosci.ki.se [130.237.109.196]) 24, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k81AMdFB020947 24, 25 -- for {microscopy-at-msa.microscopy.com} ; Fri, 1 Sep 2006 05:22:40 -0500 24, 25 -- Received: from smtp.biosci.ki.se (localhost [127.0.0.1]) 24, 25 -- by localhost (Postfix) with ESMTP id E5D124FF01; 24, 25 -- Fri, 1 Sep 2006 12:22:36 +0200 (CEST) 24, 25 -- Received: from whale.csb.ki.se (whale.csb.ki.se [130.237.109.8]) 24, 25 -- by smtp.biosci.ki.se (Postfix) with SMTP id D5CF74FEF5; 24, 25 -- Fri, 1 Sep 2006 12:22:36 +0200 (CEST) 24, 25 -- Received: from rapana.csb.ki.se by whale.csb.ki.se (5.65v4.0/1.1.10.5/03Feb98-0237PM) 24, 25 -- id AA27600; Fri, 1 Sep 2006 12:22:36 +0200 24, 25 -- Message-Id: {10609011022.AA27600-at-whale.csb.ki.se} 24, 25 -- From: "Philip Koeck" {Philip.Koeck-at-biosci.ki.se} 24, 25 -- To: {kevin-at-mediacy.com} , {microscopy-at-msa.microscopy.com} 24, 25 -- Subject: RE: [Microscopy] Question: pixels, pixels, little pixels.... 24, 25 -- Date: Fri, 1 Sep 2006 12:22:33 +0200 24, 25 -- Mime-Version: 1.0 24, 25 -- Content-Type: text/plain; 24, 25 -- charset="us-ascii" 24, 25 -- Content-Transfer-Encoding: 7bit 24, 25 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 24, 25 -- In-Reply-To: {200608311405.k7VE5xD3019349-at-ns.microscopy.com} 24, 25 -- X-Mimeole: Produced By Microsoft MimeOLE V6.00.2900.2962 24, 25 -- Thread-Index: AcbNBpYyXEQCHSOMQbWzzDoUFOvEoAAn1JHg ==============================End of - Headers==============================
In order to get measure of perimeter the image analyser software must be able to recognise a corner as distinct from a horizontal or vertical row of pixels, and apply a weighting factor when it encounters a corner (otherwise a D shape would give the same perimeter as an O shape if perimeter is calculated merely by pixel count).
However, even more importantly, perimeter is a measurement that truly varies with magnification, and hence resolution, of the measuring conditions. If you measure lung tissue alveolar wall area & perimeter by light microscopy and then by TEM, you will get roughly the same area measurement for both, but the perimeter value will be far higher. This is because under very high magnification you will encounter further tiny small scale convolutions that mirror the larger ones seen under light microscopy. Likewise if you measure the perimeter of UK on a map, you will naturally get an incredibly small perimeter value compared the very very large value you would measure if you actually walked round all of the coastline with a treadmill.
Object area can vary also with magnification as well, if the lower magnification fails to resolve many small objects amongst larger ones, but even in this case the area differences due to magnification would still be relatively small.
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: frank.karl-at-degussa.com [mailto:frank.karl-at-degussa.com] Sent: 30 August 2006 16:30 To: keith.morris-at-ucl.ac.uk
Hello Everyone, Recently I been in a discussion about how many pixels a feature should contain to provide meaningful results from image analysis. For example, if I threshold an image or measure a perimeter how many pixels do I need a feature to have as to insure I have "statistical meaningful" data. It seem intuitive that I should have as many as possible, but what about a particle or feature that has only 12 pixels maximum in one direction (say a fiber)?
I realize I could have a rectangle 9 by 7 pixels which would give me a diagonal of 11 pixels, but if I could only measure features that had at least 10 pixels would this feature have meaning?
I think I've expressed myself as clearly as my current understand is will allow me, I'll turn this question loose on the high tension thinkers out there!
Thanks
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
This e-mail transmission, and any documents, files or previous e-mail messages attached to it may contain information that is confidential or legally privileged. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that you must not read this transmission and that any disclosure, copying, printing, distribution or use of any of the information contained in or attached to this transmission is STRICTLY PROHIBITED. If you have received this transmission in error, please immediately notify the sender by telephone or return e-mail and delete the original transmission and its attachments without reading or saving in any manner. Thank you.
==============================Original Headers============================== 10, 17 -- From frank.karl-at-degussa.com Wed Aug 30 10:24:53 2006 10, 17 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 10, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7UFOp01018353 10, 17 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 10:24:52 -0500 10, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 10, 17 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k7UFOhD0022271 10, 17 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 17:24:49 +0200 10, 17 -- Subject: Question: pixels, pixels, little pixels.... 10, 17 -- To: microscopy-at-msa.microscopy.com 10, 17 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 10, 17 -- Message-ID: {OF10988FCD.61D70032-ON852571DA.0052EAEE-852571DA.0054A7DA-at-degussa.com} 10, 17 -- From: frank.karl-at-degussa.com 10, 17 -- Date: Wed, 30 Aug 2006 11:24:39 -0400 10, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 10, 17 -- 08/30/2006 10:24:49 AM 10, 17 -- MIME-Version: 1.0 10, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
==============================Original Headers============================== 29, 26 -- From keith.morris-at-ucl.ac.uk Fri Sep 1 06:21:53 2006 29, 26 -- Received: from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 29, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k81BLqv1031824 29, 26 -- for {Microscopy-at-microscopy.com} ; Fri, 1 Sep 2006 06:21:52 -0500 29, 26 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 29, 26 -- by vscani-e.ucl.ac.uk with esmtp (Exim 4.60) 29, 26 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 29, 26 -- id 1GJ75m-0003to-0c 29, 26 -- for Microscopy-at-microscopy.com; Fri, 01 Sep 2006 12:21:46 +0100 29, 26 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 29, 26 -- To: {Microscopy-at-microscopy.com} 29, 26 -- Subject: RE: [Microscopy] Question: pixels, pixels, little pixels.... 29, 26 -- Date: Fri, 1 Sep 2006 12:21:45 +0100 29, 26 -- Message-ID: {005a01c6cdb8$cf348500$7b865290-at-keithhigrade} 29, 26 -- MIME-Version: 1.0 29, 26 -- Content-Type: text/plain; 29, 26 -- charset="us-ascii" 29, 26 -- Content-Transfer-Encoding: 7bit 29, 26 -- X-Mailer: Microsoft Office Outlook 11 29, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 29, 26 -- In-Reply-To: {200608301530.k7UFUEU3025287-at-ns.microscopy.com} 29, 26 -- Thread-Index: AcbMSTMuSc83fszEQ/GLZIAHWeHOYgBay+Uw 29, 26 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 29, 26 -- X-UCL-MailScanner: Found to be clean 29, 26 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 29, 26 -- X-Spam-Status: No ==============================End of - Headers==============================
Mark, If you can move the cabinets away from the wall, even a half inch, the vibrations should lessen considerably. Pat Connelly NIH/NHLBI Bethesda, MD connellyps-at-mail.nih.gov
-----Original Message----- X-from: mbush-at-fit.edu [mailto:mbush-at-fit.edu] Sent: Thursday, August 31, 2006 6:53 PM To: Connelly, Patricia (NIH/NHLBI) [E]
Hi Keith,
The fact that the length of, say the coast of the UK, depends on the scale you are using is known as "fractal dimansion". A lot of work has been done on fractal dimensions, and it is not limited to digital images.
Regarding the original question about errors and reliability of measurements of digital images, I think that the answer is determined by applying the Nyquist-Shannon theorem to digital images. There is actually a very good explanation of this in Wikipedia (for the mathematically inclined):
If you read the article, it says that this theorem is valid for certain types of signals (band limited, "infinite sample", etc.) and that there are several practical considerations that need to be taken into account for real world signals and may require some oversampling:
"The sampling theorem does not say what happens when the conditions and procedures are not exactly met, but its proof suggests an analytical framework in which the non-ideality can be studied. A designer of a system that deals with sampling and reconstruction processes needs a thorough understanding of the signal to be sampled, in particular its frequency content, the sampling frequency, how the signal is reconstructed in terms of interpolation, and the requirement for the total reconstruction error, including aliasing and interpolation error. These properties and parameters may need to be carefully tuned in order to obtain a useful system."
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: keith.morris-at-ucl.ac.uk [mailto:keith.morris-at-ucl.ac.uk] Sent: Friday, September 01, 2006 05:27 To: Mike Bode
Hi Frank,
In order to get measure of perimeter the image analyser software must be able to recognise a corner as distinct from a horizontal or vertical row of pixels, and apply a weighting factor when it encounters a corner (otherwise a D shape would give the same perimeter as an O shape if perimeter is calculated merely by pixel count).
However, even more importantly, perimeter is a measurement that truly varies with magnification, and hence resolution, of the measuring conditions. If you measure lung tissue alveolar wall area & perimeter by light microscopy and then by TEM, you will get roughly the same area measurement for both, but the perimeter value will be far higher. This is because under very high magnification you will encounter further tiny small scale convolutions that mirror the larger ones seen under light microscopy. Likewise if you measure the perimeter of UK on a map, you will naturally get an incredibly small perimeter value compared the very very large value you would measure if you actually walked round all of the coastline with a treadmill.
Object area can vary also with magnification as well, if the lower magnification fails to resolve many small objects amongst larger ones, but even in this case the area differences due to magnification would still be relatively small.
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: frank.karl-at-degussa.com [mailto:frank.karl-at-degussa.com] Sent: 30 August 2006 16:30 To: keith.morris-at-ucl.ac.uk
Hello Everyone, Recently I been in a discussion about how many pixels a feature should contain to provide meaningful results from image analysis. For example, if I threshold an image or measure a perimeter how many pixels do I need a feature to have as to insure I have "statistical meaningful" data. It seem intuitive that I should have as many as possible, but what about a particle or feature that has only 12 pixels maximum in one direction (say a fiber)?
I realize I could have a rectangle 9 by 7 pixels which would give me a diagonal of 11 pixels, but if I could only measure features that had at least 10 pixels would this feature have meaning?
I think I've expressed myself as clearly as my current understand is will allow me, I'll turn this question loose on the high tension thinkers out there!
Thanks
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
330-668-2235 Ext. 238
This e-mail transmission, and any documents, files or previous e-mail messages attached to it may contain information that is confidential or legally privileged. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that you must not read this transmission and that any disclosure, copying, printing, distribution or use of any of the information contained in or attached to this transmission is STRICTLY PROHIBITED. If you have received this transmission in error, please immediately notify the sender by telephone or return e-mail and delete the original transmission and its attachments without reading or saving in any manner. Thank you.
==============================Original Headers============================== 10, 17 -- From frank.karl-at-degussa.com Wed Aug 30 10:24:53 2006 10, 17 -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com [193.100.56.173]) 10, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7UFOp01018353 10, 17 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 10:24:52 -0500 10, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 10, 17 -- by malmailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k7UFOhD0022271 10, 17 -- for {microscopy-at-msa.microscopy.com} ; Wed, 30 Aug 2006 17:24:49 +0200 10, 17 -- Subject: Question: pixels, pixels, little pixels.... 10, 17 -- To: microscopy-at-msa.microscopy.com 10, 17 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 10, 17 -- Message-ID: {OF10988FCD.61D70032-ON852571DA.0052EAEE-852571DA.0054A7DA-at-degussa.com} 10, 17 -- From: frank.karl-at-degussa.com 10, 17 -- Date: Wed, 30 Aug 2006 11:24:39 -0400 10, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 10, 17 -- 08/30/2006 10:24:49 AM 10, 17 -- MIME-Version: 1.0 10, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
==============================Original Headers============================== 29, 26 -- From keith.morris-at-ucl.ac.uk Fri Sep 1 06:21:53 2006 29, 26 -- Received: from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 29, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k81BLqv1031824 29, 26 -- for {Microscopy-at-microscopy.com} ; Fri, 1 Sep 2006 06:21:52 -0500 29, 26 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 29, 26 -- by vscani-e.ucl.ac.uk with esmtp (Exim 4.60) 29, 26 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 29, 26 -- id 1GJ75m-0003to-0c 29, 26 -- for Microscopy-at-microscopy.com; Fri, 01 Sep 2006 12:21:46 +0100 29, 26 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 29, 26 -- To: {Microscopy-at-microscopy.com} 29, 26 -- Subject: RE: [Microscopy] Question: pixels, pixels, little pixels.... 29, 26 -- Date: Fri, 1 Sep 2006 12:21:45 +0100 29, 26 -- Message-ID: {005a01c6cdb8$cf348500$7b865290-at-keithhigrade} 29, 26 -- MIME-Version: 1.0 29, 26 -- Content-Type: text/plain; 29, 26 -- charset="us-ascii" 29, 26 -- Content-Transfer-Encoding: 7bit 29, 26 -- X-Mailer: Microsoft Office Outlook 11 29, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 29, 26 -- In-Reply-To: {200608301530.k7UFUEU3025287-at-ns.microscopy.com} 29, 26 -- Thread-Index: AcbMSTMuSc83fszEQ/GLZIAHWeHOYgBay+Uw 29, 26 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 29, 26 -- X-UCL-MailScanner: Found to be clean 29, 26 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 29, 26 -- X-Spam-Status: No ==============================End of - Headers==============================
==============================Original Headers============================== 44, 24 -- From Mike.Bode-at-olympus-sis.com Fri Sep 1 10:29:40 2006 44, 24 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) 44, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k81FTdZ4023800 44, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 1 Sep 2006 10:29:39 -0500 44, 24 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 44, 24 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id k81FdfSp011618 44, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 1 Sep 2006 17:39:43 +0200 44, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 44, 24 -- Content-class: urn:content-classes:message 44, 24 -- MIME-Version: 1.0 44, 24 -- Content-Type: text/plain; 44, 24 -- charset="us-ascii" 44, 24 -- Subject: RE: [Microscopy] RE: Question: pixels, pixels, little pixels.... 44, 24 -- Date: Fri, 1 Sep 2006 17:24:19 +0200 44, 24 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC944775B7-at-ms-s-gws.soft-imaging.net} 44, 24 -- In-Reply-To: {200609011126.k81BQk0x008243-at-ns.microscopy.com} 44, 24 -- X-MS-Has-Attach: 44, 24 -- X-MS-TNEF-Correlator: 44, 24 -- Thread-Topic: [Microscopy] RE: Question: pixels, pixels, little pixels.... 44, 24 -- Thread-Index: AcbNuYTkt4R1RhzqSrK3eg10Kx5MWAAH5elQ 44, 24 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 44, 24 -- To: {Microscopy-at-microscopy.com} 44, 24 -- Content-Transfer-Encoding: 8bit 44, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k81FTdZ4023800 ==============================End of - Headers==============================
The College of Microscopy will be offering the following electron microscopy short courses this fall:
October 16-20 - Scanning Electron Microscopy
October 31-November 2 - Transmission Electron Microscopy
Both will be held at our Westmont, IL facility. In addition to lectures, these courses emphasize hands-on training using state of the art equipment. For further details and registration information, please follow the link below.
www.collegeofmicroscopy.com
Elaine Schumacher Senior Research Scientist McCrone Associates, Inc. 850 Pasquinelli Drive Westmont, IL 60559-5539 USA 630-887-7100 (tel) 630-887-7417 (fax) E-mail: eschumacher-at-mccrone.com Web Site: www.mccrone.com
==============================Original Headers============================== 10, 27 -- From eschumacher-at-mccrone.com Fri Sep 1 10:42:19 2006 10, 27 -- Received: from pgp.mccrone.com (McCrone-Associates-1051626.cust-rtr.ameritech.net [68.23.63.122]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k81FgJUp001818 10, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 1 Sep 2006 10:42:19 -0500 10, 27 -- Received: from pgp.mccrone.com (localhost [127.0.0.1]) 10, 27 -- by pgp.mccrone.com (Postfix) with ESMTP id 536B81A8016 10, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 1 Sep 2006 10:42:21 -0500 (CDT) 10, 27 -- Received: from MCCRONEMSG.tmg.mccrone.com ([192.168.101.20]) 10, 27 -- by pgp.mccrone.com (PGP Universal service); 10, 27 -- Fri, 01 Sep 2006 10:42:21 -0500 10, 27 -- X-PGP-Universal: processed 10, 27 -- Content-class: urn:content-classes:message 10, 27 -- MIME-Version: 1.0 10, 27 -- Content-Type: text/plain; 10, 27 -- charset="us-ascii" 10, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 27 -- Subject: Short Course Announcement: SEM and TEM 10, 27 -- Date: Fri, 1 Sep 2006 10:42:09 -0500 10, 27 -- Message-ID: {2A27CE0EC2A5C748974EA513DFB285A7B9726A-at-MCCRONEMSG.tmg.mccrone.com} 10, 27 -- X-MS-Has-Attach: 10, 27 -- X-MS-TNEF-Correlator: 10, 27 -- Thread-Topic: Short Course Announcement: SEM and TEM 10, 27 -- Thread-Index: AcbN3S9BVrtsxq9mRSGihzBsZXb45g== 10, 27 -- From: "Elaine F. Schumacher" {eschumacher-at-mccrone.com} 10, 27 -- To: {Microscopy-at-MSA.Microscopy.Com} 10, 27 -- Content-Transfer-Encoding: 8bit 10, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k81FgJUp001818 ==============================End of - Headers==============================
If possible, it might be interesting to try the following:
In a circle, put three golf-balls in such a way that they touch each other and cannot move away. In the middle of these three, put a golf-ball on top of the three balls, as if you create a pyramid of golf-balls. Do this for three / four mounting places and mount the table / microscope stand on the top-balls. You'd be amazed how well golf-balls can absorb vibrations.
If the equipment is not too heavy, you can also consider tennis-balls, however, they will lift the table with about 5-6 inches, whereas the golf-balls will only add about 3 inches...
Obviously in this way you reduce the ergonomics by putting the microscope higher, but you'll also reduce the vibrations. Hope it works out, for me it did a few years ago. Best,
Sven Terclavers
-----Original Message----- X-from: mbush-at-fit.edu [mailto:mbush-at-fit.edu] Sent: Thursday, August 31, 2006 6:53 PM To: Connelly, Patricia (NIH/NHLBI) [E]
Dear listers,
I have some basic questions about the observation of minerals on nickel grids in TEM. I use formvar-carbon-coated nickel grids (I would have prefered copper but we have nickel grids in stock) and I deposit fine powder of crystal mineral onto them. When I observe the particles, even with the diffraction aperture inserted, the surface illuminated by the beam darkens pretty quickly. Sometimes I even have dark rings imprinted on the coating film (if I leave the beam for some minutes). If I take the diffrac. aperture out of the way for EDX analysis the surface becomes quickly complety black (in a matter of minutes).
1) What is black? Does the mineral melt? 2) If I heat the mineral to make it amorphous before the observation I sometimes still see dark rings. Are they diffraction rings? (in normal mode, not diffraction mode). Is there any diffraction in amorphous material? 3) In EDX I see a peak for copper, just after the nickel peak (at 8,070 keV) and I don’t expect copper in my material. Is there copper in the nickel grids?
Regards,
Stephane
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Email: ludo.rossou-at-ua.ac.be Name: Ludo Rossou
Organization: University of Antwerp Belgium
Title-Subject: [Filtered] specimen polishing
Question: Does anyone have some experience with the MULTIPREP SYSTEM of Allied High Tech Products?
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Email: Henrik.Kaker-at-guest.arnes.si Name: Henrik Kaker
The black is most likely carbonaceous contamination. Volatile organic molecules diffuse across the sample surface under the influence of the beam's electric field. When they hit the beam they "polymerize" and form a thick layer which appears dark in the image. In TEM, the contamination layer usually appears as a ring around the perimeter of the beam. As you decrease the size of the beam (e.g. STEM), the contamination problem increases.
} 2) If I heat the mineral to make it amorphous before } the observation I sometimes still see dark rings. Are } they diffraction rings? (in normal mode, not } diffraction mode). Is there any diffraction in } amorphous material?
You will see diffraction rings from amorphous material. They will be diffuse, not sharp like the rings from most crystalline materials. The carbonaceous contamination will also give rise to diffuse diffraction rings
} 3) In EDX I see a peak for copper, just after the } nickel peak (at 8,070 keV) and I don't expect copper } in my material. Is there copper in the nickel grids?
My suspicion is that you are seeing the Ni K beta peak,although the Ni Kbeta is at 8.265keV. The K beta is ~20% of the intensity of the K alpha peak. The 1st row transition metal K lines have the characteristic that an element Z's K beta falls under the (Z+1) K alpha.
At 04:25 AM 9/4/2006, you wrote:
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Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
==============================Original Headers============================== 14, 26 -- From colijn.1-at-osu.edu Mon Sep 4 14:43:05 2006 14, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 14, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k84Jh5tI023115 14, 26 -- for {microscopy-at-microscopy.com} ; Mon, 4 Sep 2006 14:43:05 -0500 14, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 14, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 14, 26 -- id {01M6SWBB13M8A8Z7NB-at-er6s1.eng.ohio-state.edu} for 14, 26 -- microscopy-at-microscopy.com; Mon, 04 Sep 2006 15:43:04 -0400 (EDT) 14, 26 -- Received: from HOC3.osu.edu 14, 26 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 14, 26 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 14, 26 -- with ESMTPA id {01M6SWBABN76A8NJ85-at-er6s1.eng.ohio-state.edu} ; Mon, 14, 26 -- 04 Sep 2006 15:43:03 -0400 (EDT) 14, 26 -- Date: Mon, 04 Sep 2006 15:39:23 -0400 14, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 14, 26 -- Subject: Re: [Microscopy] basic questions about Ni grids 14, 26 -- In-reply-to: {200609040825.k848PxJd001086-at-ns.microscopy.com} 14, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 14, 26 -- To: nizets2-at-yahoo.com 14, 26 -- Cc: microscopy-at-microscopy.com 14, 26 -- Message-id: {7.0.1.0.2.20060904145709.024ed3a8-at-osu.edu} 14, 26 -- MIME-version: 1.0 14, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 14, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 14, 26 -- References: {200609040825.k848PxJd001086-at-ns.microscopy.com} ==============================End of - Headers==============================
Stephane, I agree with Henk that it is probably contamination on your samples. To make sure, if you tilt the samples at a high angle, you should see a split of the rings where you are seeing the top and bottom surfaces. This used to be a way of measuring the thickness of a TEM sample. You can download a copy of the paper from our website on contamination of samples. The URL is http://www.southbaytech.com/app_index.cfm?main_action=tech_papers" and the title is "Surface Science Aspects of Contamination in TEM Sample Prep" by John Grant et al. and it is paper number 225. (I am one of the authors.) In that paper, you will see a tilted sample with heavy contamination. To avoid contamination like this, you should plasma clean your sample prior to putting it into the TEM.
Disclaimer: South Bay Technology makes and sells a Plasma Cleaner.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
-------- Original Message -------- } From: colijn.1-at-osu.edu } Sent: Monday, September 04, 2006 3:47 PM } To: Walck-at-SouthBayTech.com } Subject: [Microscopy] Re: basic questions about Ni 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 } ---------------------------------------------------------------------------- } } Stephane, } } } 1) What is black? Does the mineral melt? } } The black is most likely carbonaceous contamination. Volatile } organic molecules diffuse across the sample surface under the } influence of the beam's electric field. When they hit the beam they } "polymerize" and form a thick layer which appears dark in the } image. In TEM, the contamination layer usually appears as a ring } around the perimeter of the beam. As you decrease the size of the } beam (e.g. STEM), the contamination problem increases. } } } 2) If I heat the mineral to make it amorphous before } } the observation I sometimes still see dark rings. Are } } they diffraction rings? (in normal mode, not } } diffraction mode). Is there any diffraction in } } amorphous material? } } You will see diffraction rings from amorphous material. They will be } diffuse, not sharp like the rings from most crystalline } materials. The carbonaceous contamination will also give rise to } diffuse diffraction rings } } } } 3) In EDX I see a peak for copper, just after the } } nickel peak (at 8,070 keV) and I don't expect copper } } in my material. Is there copper in the nickel grids? } } My suspicion is that you are seeing the Ni K beta peak,although the } Ni Kbeta is at 8.265keV. The K beta is ~20% of the intensity of the } K alpha peak. The 1st row transition metal K lines have the } characteristic that an element Z's K beta falls under the (Z+1) K alpha. } } At 04:25 AM 9/4/2006, 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 } } ---------------------------------------------------------------------------- } } } } Dear listers, } } } } I have some basic questions about the observation of } } minerals on nickel grids in TEM. } } I use formvar-carbon-coated nickel grids (I would have } } prefered copper but we have nickel grids in stock) and } } I deposit fine powder of crystal mineral onto them. } } When I observe the particles, even with the } } diffraction aperture inserted, the surface illuminated } } by the beam darkens pretty quickly. Sometimes I even } } have dark rings imprinted on the coating film (if I } } leave the beam for some minutes). If I take the } } diffrac. aperture out of the way for EDX analysis the } } surface becomes quickly complety black (in a matter of } } minutes). } } } } 1) What is black? Does the mineral melt? } } 2) If I heat the mineral to make it amorphous before } } the observation I sometimes still see dark rings. Are } } they diffraction rings? (in normal mode, not } } diffraction mode). Is there any diffraction in } } amorphous material? } } 3) In EDX I see a peak for copper, just after the } } nickel peak (at 8,070 keV) and I don't expect copper } } in my material. Is there copper in the nickel grids? } } } } Regards, } } } } Stephane } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? Yahoo! Mail has the best spam protection around } } http://mail.yahoo.com } } } } ==============================Original Headers============================== } } 6, 18 -- From nizets2-at-yahoo.com Mon Sep 4 03:22:21 2006 } } 6, 18 -- Received: from web37409.mail.mud.yahoo.com } } (web37409.mail.mud.yahoo.com [209.191.91.141]) } } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } SMTP id k848MLdD030715 } } 6, 18 -- for {microscopy-at-microscopy.com} ; Mon, 4 Sep 2006 } } 03:22:21 -0500 } } 6, 18 -- Received: (qmail 26154 invoked by uid 60001); 4 Sep 2006 } } 08:22:21 -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=Z1dFUFwj1Z4BU2u7V3woEcOWcTaYDVkXOHEnRSHjKc9qFplAgfLdzsnVVPkb9kQF8Z7sWvp9QcJe3yWEBEzqkbybAmNyVRh8DcOuLa5I/WqtS+hDjAB+dVoRAf/2P94bZlGvet8AWydQt05cDmOsyz8pk3qZZo+OKmgbnHGRkOw= } } ; } } 6, 18 -- Message-ID: {20060904082221.26152.qmail-at-web37409.mail.mud.yahoo.com} } } 6, 18 -- Received: from [80.122.101.102] by } } web37409.mail.mud.yahoo.com via HTTP; Mon, 04 Sep 2006 01:22:21 PDT } } 6, 18 -- Date: Mon, 4 Sep 2006 01:22:21 -0700 (PDT) } } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } } 6, 18 -- Subject: basic questions about Ni grids } } 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============================== } } Hendrik O. Colijn colijn.1-at-osu.edu } OSU Campus Electron Optics Facility www.ceof.ohio-state.edu } 040 Fontana Labs, 116 W. 19th Ave } } } ==============================Original Headers============================== } 14, 26 -- From colijn.1-at-osu.edu Mon Sep 4 14:43:05 2006 } 14, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) } 14, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k84Jh5tI023115 } 14, 26 -- for {microscopy-at-microscopy.com} ; Mon, 4 Sep 2006 14:43:05 -0500 } 14, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by } 14, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) } 14, 26 -- id {01M6SWBB13M8A8Z7NB-at-er6s1.eng.ohio-state.edu} for } 14, 26 -- microscopy-at-microscopy.com; Mon, 04 Sep 2006 15:43:04 -0400 (EDT) } 14, 26 -- Received: from HOC3.osu.edu } 14, 26 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) } 14, 26 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) } 14, 26 -- with ESMTPA id {01M6SWBABN76A8NJ85-at-er6s1.eng.ohio-state.edu} ; Mon, } 14, 26 -- 04 Sep 2006 15:43:03 -0400 (EDT) } 14, 26 -- Date: Mon, 04 Sep 2006 15:39:23 -0400 } 14, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} } 14, 26 -- Subject: Re: [Microscopy] basic questions about Ni grids } 14, 26 -- In-reply-to: {200609040825.k848PxJd001086-at-ns.microscopy.com} } 14, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu } 14, 26 -- To: nizets2-at-yahoo.com } 14, 26 -- Cc: microscopy-at-microscopy.com } 14, 26 -- Message-id: {7.0.1.0.2.20060904145709.024ed3a8-at-osu.edu} } 14, 26 -- MIME-version: 1.0 } 14, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } 14, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed } 14, 26 -- X-Env-From: auth/colijn.1-at-osu.edu } 14, 26 -- References: {200609040825.k848PxJd001086-at-ns.microscopy.com} } ==============================End of - Headers==============================
==============================Original Headers============================== 5, 21 -- From walck-at-southbaytech.com Mon Sep 4 16:59:00 2006 5, 21 -- Received: from smtp14.dc2.safesecureweb.com (smtp14.dc2.safesecureweb.com [65.36.255.238]) 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k84LwxTP003026 5, 21 -- for {microscopy-at-microscopy.com} ; Mon, 4 Sep 2006 16:59:00 -0500 5, 21 -- Received: from mail43.safesecureweb.com (barrage.ad.safesecureweb.com [10.10.1.193]) 5, 21 -- by smtp14.dc2.safesecureweb.com (Spam Firewall) with ESMTP 5, 21 -- id 8CBE09A00B9; Mon, 4 Sep 2006 17:58:56 -0400 (EDT) 5, 21 -- MIME-Version: 1.0 5, 21 -- Date: Mon, 4 Sep 2006 17:58:36 -0400 5, 21 -- Received: from [222.151.229.88] by mail43.safesecureweb.com via HTTP; Mon, 4 Sep 2006 17:58:36 -0400 5, 21 -- Content-Type: text/plain; 5, 21 -- charset=iso-8859-1 5, 21 -- Subject: re: [Microscopy] Re: basic questions about Ni grids 5, 21 -- From: "Scott Walck" {walck-at-southbaytech.com} 5, 21 -- Reply-To: Walck-at-southbaytech.com 5, 21 -- To: {microscopy-at-microscopy.com} 5, 21 -- CC: {colijn.1-at-osu.edu} 5, 21 -- Message-ID: {b1b9f360d89d4fb1a0b4ff63e3f099fa-at-southbaytech.com} 5, 21 -- X-Virus-Scanned: by Barracuda Spam Firewall at safesecureweb.com 5, 21 -- Content-Transfer-Encoding: 8bit 5, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k84LwxTP003026 ==============================End of - Headers==============================
Be careful about putting a carbon coated formvar grid into an oxygen plasma cleaner. The oxygen plasma does a great job of removing carbon from your sample, including the carbon containing support film!. You may clean the support film completely away. That said, a plasma cleaner is a great thing to have available for the right type of sample and is almost a necessity for FEG STEM work.
I have often reduced (though not eliminated) carbon contamination by warming the TEM grid to ~100C just before putting it into the scope. It seems to be enough to desorb most of the organic contaminants. Obviously you should be careful to keep the microscope and sample rods absolutely clean (no fingerprints, organic solvents, etc.) Cleaning the sample rod in a plasma cleaner really helps.
Cheers, Henk
At 05:58 PM 9/4/2006, Scott Walck wrote: } Stephane, } I agree with Henk that it is probably contamination on your samples. } To make sure, if you tilt the samples at a high angle, you should } see a split of the rings where you are seeing the top and bottom surfaces. } This used to be a way of measuring the thickness of a TEM sample. } You can download a copy of the paper from our website on } contamination of samples. The URL is } http://www.southbaytech.com/app_index.cfm?main_action=tech_papers" } and the title is "Surface Science Aspects of Contamination in TEM } Sample Prep" by John Grant et al. and it is paper number 225. (I } am one of the authors.) In that paper, you will } see a tilted sample with heavy contamination. To avoid } contamination like this, you should plasma clean your sample prior } to putting it into the TEM. } } Disclaimer: South Bay Technology makes and sells a Plasma Cleaner. } } -Scott } } Scott D. Walck, Ph.D. } Technical Director } South Bay Technology, Inc. } 1120 Via Callejon } San Clemente, CA 92673 } } TEL: +1-949-492-2600 } FAX: +1-949-492-1499 } walck-at-southbaytech.com } } -------- Original Message -------- } } From: colijn.1-at-osu.edu } } Sent: Monday, September 04, 2006 3:47 PM } } To: Walck-at-SouthBayTech.com } } Subject: [Microscopy] Re: basic questions about Ni 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 } } } ---------------------------------------------------------------------------- } } } } Stephane, } } } } } 1) What is black? Does the mineral melt? } } } } The black is most likely carbonaceous contamination. Volatile } } organic molecules diffuse across the sample surface under the } } influence of the beam's electric field. When they hit the beam they } } "polymerize" and form a thick layer which appears dark in the } } image. In TEM, the contamination layer usually appears as a ring } } around the perimeter of the beam. As you decrease the size of the } } beam (e.g. STEM), the contamination problem increases. } } } } } 2) If I heat the mineral to make it amorphous before } } } the observation I sometimes still see dark rings. Are } } } they diffraction rings? (in normal mode, not } } } diffraction mode). Is there any diffraction in } } } amorphous material? } } } } You will see diffraction rings from amorphous material. They will be } } diffuse, not sharp like the rings from most crystalline } } materials. The carbonaceous contamination will also give rise to } } diffuse diffraction rings } } } } } } } 3) In EDX I see a peak for copper, just after the } } } nickel peak (at 8,070 keV) and I don't expect copper } } } in my material. Is there copper in the nickel grids? } } } } My suspicion is that you are seeing the Ni K beta peak,although the } } Ni Kbeta is at 8.265keV. The K beta is ~20% of the intensity of the } } K alpha peak. The 1st row transition metal K lines have the } } characteristic that an element Z's K beta falls under the (Z+1) K alpha. } } } } At 04:25 AM 9/4/2006, 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 } } } ----------------------------------------------------------------- } ----------- } } } } } } Dear listers, } } } } } } I have some basic questions about the observation of } } } minerals on nickel grids in TEM. } } } I use formvar-carbon-coated nickel grids (I would have } } } prefered copper but we have nickel grids in stock) and } } } I deposit fine powder of crystal mineral onto them. } } } When I observe the particles, even with the } } } diffraction aperture inserted, the surface illuminated } } } by the beam darkens pretty quickly. Sometimes I even } } } have dark rings imprinted on the coating film (if I } } } leave the beam for some minutes). If I take the } } } diffrac. aperture out of the way for EDX analysis the } } } surface becomes quickly complety black (in a matter of } } } minutes). } } } } } } 1) What is black? Does the mineral melt? } } } 2) If I heat the mineral to make it amorphous before } } } the observation I sometimes still see dark rings. Are } } } they diffraction rings? (in normal mode, not } } } diffraction mode). Is there any diffraction in } } } amorphous material? } } } 3) In EDX I see a peak for copper, just after the } } } nickel peak (at 8,070 keV) and I don't expect copper } } } in my material. Is there copper in the nickel grids? } } } } } } Regards, } } } } } } Stephane } } } } } } __________________________________________________ } } } Do You Yahoo!? } } } Tired of spam? Yahoo! Mail has the best spam protection around } } } http://mail.yahoo.com } } } } } } ==============================Original } Headers============================== } } } 6, 18 -- From nizets2-at-yahoo.com Mon Sep 4 03:22:21 2006 } } } 6, 18 -- Received: from web37409.mail.mud.yahoo.com } } } (web37409.mail.mud.yahoo.com [209.191.91.141]) } } } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } } SMTP id k848MLdD030715 } } } 6, 18 -- for {microscopy-at-microscopy.com} ; Mon, 4 Sep 2006 } } } 03:22:21 -0500 } } } 6, 18 -- Received: (qmail 26154 invoked by uid 60001); 4 Sep 2006 } } } 08:22:21 -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-T } ype:Content-Transfer-Encoding; } } } 6, 18 } } } -- } } } b=Z1dFUFwj1Z4BU2u7V3woEcOWcTaYDVkXOHEnRSHjKc9qFplAgfLdzsnVVPkb9kQ } F8Z7sWvp9QcJe3yWEBEzqkbybAmNyVRh8DcOuLa5I/WqtS+hDjAB+dVoRAf/2P94bZlGvet8AWydQt05cDmOsyz8pk3qZZo+OKmgbnHGRkOw= } } } } ; } } } 6, 18 -- Message-ID: } {20060904082221.26152.qmail-at-web37409.mail.mud.yahoo.com} } } } 6, 18 -- Received: from [80.122.101.102] by } } } web37409.mail.mud.yahoo.com via HTTP; Mon, 04 Sep 2006 01:22:21 PDT } } } 6, 18 -- Date: Mon, 4 Sep 2006 01:22:21 -0700 (PDT) } } } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } } } 6, 18 -- Subject: basic questions about Ni grids } } } 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============================== } } } } Hendrik O. Colijn colijn.1-at-osu.edu } } OSU Campus Electron Optics Facility www.ceof.ohio-state.edu } } 040 Fontana Labs, 116 W. 19th Ave } } } } } } ==============================Original } Headers============================== } } 14, 26 -- From colijn.1-at-osu.edu Mon Sep 4 14:43:05 2006 } } 14, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU } (er6s1.ecr6.ohio-state.edu [164.107.76.2]) } } 14, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k84Jh5tI023115 } } 14, 26 -- for {microscopy-at-microscopy.com} ; Mon, 4 Sep 2006 } 14:43:05 -0500 } } 14, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by } } 14, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) } } 14, 26 -- id {01M6SWBB13M8A8Z7NB-at-er6s1.eng.ohio-state.edu} for } } 14, 26 -- microscopy-at-microscopy.com; Mon, 04 Sep 2006 15:43:04 -0400 (EDT) } } 14, 26 -- Received: from HOC3.osu.edu } } 14, 26 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) } } 14, 26 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) } } 14, 26 -- with ESMTPA id } {01M6SWBABN76A8NJ85-at-er6s1.eng.ohio-state.edu} ; Mon, } } 14, 26 -- 04 Sep 2006 15:43:03 -0400 (EDT) } } 14, 26 -- Date: Mon, 04 Sep 2006 15:39:23 -0400 } } 14, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} } } 14, 26 -- Subject: Re: [Microscopy] basic questions about Ni grids } } 14, 26 -- In-reply-to: {200609040825.k848PxJd001086-at-ns.microscopy.com} } } 14, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu } } 14, 26 -- To: nizets2-at-yahoo.com } } 14, 26 -- Cc: microscopy-at-microscopy.com } } 14, 26 -- Message-id: {7.0.1.0.2.20060904145709.024ed3a8-at-osu.edu} } } 14, 26 -- MIME-version: 1.0 } } 14, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } } 14, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed } } 14, 26 -- X-Env-From: auth/colijn.1-at-osu.edu } } 14, 26 -- References: {200609040825.k848PxJd001086-at-ns.microscopy.com} } } ==============================End of - } Headers==============================
Hendrik O. Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility www.ceof.ohio-state.edu 040 Fontana Labs, 116 W. 19th Ave
==============================Original Headers============================== 9, 25 -- From colijn.1-at-osu.edu Mon Sep 4 19:24:45 2006 9, 25 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 9, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k850Oj5Y015415 9, 25 -- for {microscopy-at-microscopy.com} ; Mon, 4 Sep 2006 19:24:45 -0500 9, 25 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 9, 25 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 9, 25 -- id {01M6T65J6080A8ZTI9-at-er6s1.eng.ohio-state.edu} for 9, 25 -- microscopy-at-microscopy.com; Mon, 04 Sep 2006 20:24:44 -0400 (EDT) 9, 25 -- Received: from HOC3.osu.edu 9, 25 -- (d149-67-33-174.col.wideopenwest.com [67.149.174.33]) 9, 25 -- by er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 9, 25 -- with ESMTPA id {01M6T65IDHM2A90YS3-at-er6s1.eng.ohio-state.edu} ; Mon, 9, 25 -- 04 Sep 2006 20:24:44 -0400 (EDT) 9, 25 -- Date: Mon, 04 Sep 2006 20:24:25 -0400 9, 25 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 9, 25 -- Subject: re: [Microscopy] Re: basic questions about Ni grids 9, 25 -- In-reply-to: {b1b9f360d89d4fb1a0b4ff63e3f099fa-at-southbaytech.com} 9, 25 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 9, 25 -- To: nizets2-at-yahoo.com, Walck-at-southbaytech.com, microscopy-at-microscopy.com 9, 25 -- Message-id: {7.0.1.0.2.20060904200959.0231d830-at-osu.edu} 9, 25 -- MIME-version: 1.0 9, 25 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 25 -- Content-type: text/plain; charset=us-ascii; format=flowed 9, 25 -- X-Env-From: auth/colijn.1-at-osu.edu 9, 25 -- References: {b1b9f360d89d4fb1a0b4ff63e3f099fa-at-southbaytech.com} ==============================End of - Headers==============================
As no-one seems to have replied to your post, I'll pass on my thoughts and experience now I'm back from my hols.
I don't have the professional experience with video I have with microscope and SLR camera systems (on microscopes even our 'video' is essentially multi-tiff time-lapse photographic sequences rather than MP4, VOB etc. However I am a regular user of Adobe Premiere for video compression and home use, plus I now use DVD regularly to backup film and vinyl LP HIFI audio (to MP3).
I have an elderly 'consumer' Hi8 (PAL though) and now MiniDV camcorders at home (not a pro Hi8 granted). Despite investing in Adobe Premiere Pro, various ATI AGP graphic AD converters (for Hi8) and USB2 streaming/firewire for MiniDV, with transfer to a PC I always seem to get 'compression' effects. This is a quite poor video image after capturing to the PC, editing and writing to DVD, compared to what I get just feeding the image directly into a TV via the camera's inbuilt AV out (when the image quality is excellent). This poorer image quality is evident whether on the PC VDU or on viewing on a TV via DVD players after PC writing. Granted (other than Adobe Premiere that came via work), I have done this on the cheap compared to pro TV recording studio systems, being my money. However the PC always seems to convert the edited video to noticeably compressed formats, and it showed.
However if I ignore the PC and simply feed the camcorder straight into my high quality TV Panasonic DVD recorder via analogue AV3 and record to -R DVD's at XP (1h super quality mode), I get always get a superb picture as good as 610 lines PAL. This is despite going though the MiniDV camcorder and DVD recorder AD converters, and being fed out via the standard red/black/yellow phono plugs. I know NTSC has a different 'analogue out' to my PAL, but I'm sure it will work as well using the NTSC analogue into the TV DVD recorder. Besides image quality on Hi8 isn't that great compared to MiniDV (~20% better pixel quality), and it below even normal PAL resolution.
It may be that TV just shows video better than my PC CRT's but video written via Adobe Premiere and Nero from PC digital capture always looks far far worse than the direct to DVD recorder method (I just can't get rid of compression effects even if ask for super duper plus quality). It's possible that a TV DVD recorder is just really good at one thing and the PC is a bit worse at many things. Also writing to DVD is real-time via the DVD recorder, as you play the tape, it records the DVD. I assume you can edit the standard DVD-R video (once finalised) on the PC if you want afterwards. Viewing the DVD is obviously no problem on a DVD player.
Anyway give it a try if you have a spare quality TV DVD recorder (mine's a Panasonic for £250). Just avoid DVD RAM and stick to finalised DVD-R's (or +R's). I'm sure you could get better with expensive professional analogue to digital converters etc, but given your source is an NTSC (Never The Same Colour twice) old analogue Hi8 tape perhaps markedly better isn't really possible anyway. Modern TV DVD recorders can also have a digital Firewire in port as well s analogue if you use MiniDV. Digital 'walkman's' seem expensive for what they offer (ie. portability).
I have read letters into PC Pro saying the same as above and the editor hasn't offered a better alternative. Your 'time lapse' motion capture does complicate the issue though, as cheap video editing on the PC will probably bring in compression problems again.
I note that Hi8 video tape cassettes have a projected serviceable life-time of 10 years before the plastic components of the tape guide start to fail, plus the tape will be Deteriorating each year due to print-through etc..(fast forward/rewind them occasionally), so now is the time for all of us to archive to DVD.
Hope this helps.
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: edelmare-at-muohio.edu [mailto:edelmare-at-muohio.edu] Sent: 22 August 2006 21:15 To: keith.morris-at-ucl.ac.uk
Looking to digitize analog NTSC video tape (Hi-8 and VHS, so composite video, not S-video or component). Would like to stream the analog signal right in to digital format for later image analysis (so only need the video data no sound), and would like to work whole 60min tapes at a time. Since we’re looking to collect measurement data from video still images the higher the quality preserved the better. So looking for *specific* recommend ations as to the best method for doing this (specific as in model numbers).
Computer system (Windows PC) we have for this: has 1394, USB, 16x PCIe, x4 PCIe, 64-bit PCI, and 32-Bit PCI, with a RAID Array. Adobe Premiere reports system can sustain over 35MB/sec throughput.
Possible solutions:
1) A/D capture card, o.k., which one for running under Windows XP? Capturing composite NTSC video seems to be “old-tech” these days. Seemingly easy to find HDTV capture cards but not NTSC.
2) A/D capture “widgets” = Composite video in and IEEE 1394 output or USB? Seems like a very cost effective solution, can anyone recommend a model?
3) Using either a Digital8 camcorder or something like a Sony Digital8 Video Walkman to play the “tapes” out to 1394 and input the 1394 to the computer. (Note: Digital8 Players will play Hi-8 tapes). Will this work?
Video / Image Analysis: Once digitized the “video” will be broken up into digital stills (yes, 18 hours of still images at 29 frames / sec.). We actually only need 1 image from approximately every 4 seconds of video. Every 4 seconds the video image changes to one of 16 “views”. We will then sort the images into the individual 16 “views”. Each of the 16 “views” shows 7 developing seedlings. We will then analyze each of the 16 “views” (or sets of seedlings) for growth changes through time. Oh, yes after each set of 16 “views” the tape is “paused” for 10 mins, and then repeats the 16 view capture sequence again. So basically it is time lapse imaging, of 16 different subjects, all interleaved on one video stream.
And why collect data this way you ask? Because the data collection is all done automatically and remotely . . . From a minimum of 355 miles up in orbit on the International Space Station, on an automated growth and imaging system, which does not have room for 16 video cameras and video capture systems.
Any suggestions would be greatly appreciated.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 16, 24 -- From edelmare-at-muohio.edu Tue Aug 22 15:12:24 2006 16, 24 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MKCNVa013909 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 15:12:24 -0500 16, 24 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 16, 24 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7MKCBHt014555 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 16:12:12 -0400 16, 24 -- Received: from emf03 ([134.53.14.97]) 16, 24 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7MKC9EJ014949 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 16:12:09 -0400 16, 24 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 16, 24 -- To: microscopy-at-Microscopy.com 16, 24 -- Date: Tue, 22 Aug 2006 16:14:11 -0400 16, 24 -- MIME-Version: 1.0 16, 24 -- Content-type: text/plain; charset=ISO-8859-1 16, 24 -- Subject: Looking for some video capture recommendations 16, 24 -- Message-ID: {44EB2D53.3411.1471D88D-at-localhost} 16, 24 -- Priority: normal 16, 24 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 16, 24 -- X-Real-ConnectIP: 134.53.14.97 16, 24 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.67 16, 24 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 16, 24 -- Content-Transfer-Encoding: 8bit 16, 24 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k7MKCNVa013909 ==============================End of - Headers==============================
==============================Original Headers============================== 37, 27 -- From keith.morris-at-ucl.ac.uk Tue Sep 5 04:23:01 2006 37, 27 -- Received: from vscani-a.ucl.ac.uk (vscani-a.ucl.ac.uk [144.82.108.29]) 37, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k859N1IH001685 37, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Sep 2006 04:23:01 -0500 37, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 37, 27 -- by vscani-a.ucl.ac.uk with esmtp (Exim 4.60) 37, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 37, 27 -- id 1GKX8y-0001Cc-F2; Tue, 05 Sep 2006 10:22:56 +0100 37, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 37, 27 -- To: {edelmare-at-muohio.edu} 37, 27 -- Cc: {Microscopy-at-microscopy.com} 37, 27 -- Subject: RE: [Microscopy] Looking for some video capture recommendations 37, 27 -- Date: Tue, 5 Sep 2006 10:22:56 +0100 37, 27 -- Message-ID: {001801c6d0cc$df5a4a20$7b865290-at-keithhigrade} 37, 27 -- MIME-Version: 1.0 37, 27 -- Content-Type: text/plain; 37, 27 -- charset="iso-8859-1" 37, 27 -- X-Mailer: Microsoft Office Outlook 11 37, 27 -- Thread-Index: AcbGJ6PBZ9kiWw2FQpaHHhgCHiyF0AAm8PIQ 37, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 37, 27 -- In-Reply-To: {200608222014.k7MKErv0020837-at-ns.microscopy.com} 37, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 37, 27 -- X-UCL-MailScanner: Found to be clean 37, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 37, 27 -- X-Spam-Status: No 37, 27 -- Content-Transfer-Encoding: 8bit 37, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k859N1IH001685 ==============================End of - Headers==============================
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Title-Subject: [Filtered] Using RGB filters with monochrome camera
Question: Dear Listers
I intend using a high resolution (Basler) monochrome camera on a Leica DMLM materials microscope with epi illumination. If I take successive images using red, green & blue filters in turn (mounted in the filter magazine between lamp & cube), can I then assemble good quality colour images using something like ImageJ? I notice that there are a few papers on the technique but I'm not clear where it's possible to use the colour filters.
I have had good luck with a Plextor ConvertX PX-M402U. Got it a couple years ago for around $130. Not sure if they still make it. It outputs in several formats and resolutions including mpeg, divx etc. You can copy to a hard drive or burn a DVD on the fly. RCA video and audio in, USB2 data out.
Regards,
Tom
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Looking to digitize analog NTSC video tape (Hi-8 and VHS, so composite video, not S-video or component). Would like to stream the analog signal right in to digital format for later image analysis (so only need the video data no sound), and would like to work whole 60min tapes at a time. Since we're looking to collect measurement data from video still images the higher the quality preserved the better. So looking for *specific* recommend ations as to the best method for doing this (specific as in model numbers).
Computer system (Windows PC) we have for this: has 1394, USB, 16x PCIe, x4 PCIe, 64-bit PCI, and 32-Bit PCI, with a RAID Array. Adobe Premiere reports system can sustain over 35MB/sec throughput.
Possible solutions:
1) A/D capture card, o.k., which one for running under Windows XP? Capturing composite NTSC video seems to be "old-tech" these days. Seemingly easy to find HDTV capture cards but not NTSC.
2) A/D capture "widgets" = Composite video in and IEEE 1394 output or USB? Seems like a very cost effective solution, can anyone recommend a model?
3) Using either a Digital8 camcorder or something like a Sony Digital8 Video Walkman to play the "tapes" out to 1394 and input the 1394 to the computer. (Note: Digital8 Players will play Hi-8 tapes). Will this work?
Video / Image Analysis: Once digitized the "video" will be broken up into digital stills (yes, 18 hours of still images at 29 frames / sec.). We actually only need 1 image from approximately every 4 seconds of video. Every 4 seconds the video image changes to one of 16 "views". We will then sort the images into the individual 16 "views". Each of the 16 "views" shows 7 developing seedlings. We will then analyze each of the 16 "views" (or sets of seedlings) for growth changes through time. Oh, yes after each set of 16 "views" the tape is "paused" for 10 mins, and then repeats the 16 view capture sequence again. So basically it is time lapse imaging, of 16 different subjects, all interleaved on one video stream.
And why collect data this way you ask? Because the data collection is all done automatically and remotely . . . From a minimum of 355 miles up in orbit on the International Space Station, on an automated growth and imaging system, which does not have room for 16 video cameras and video capture systems.
Any suggestions would be greatly appreciated.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 16, 24 -- From edelmare-at-muohio.edu Tue Aug 22 15:12:24 2006 16, 24 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k7MKCNVa013909 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 15:12:24 -0500 16, 24 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 16, 24 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7MKCBHt014555 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 16:12:12 -0400 16, 24 -- Received: from emf03 ([134.53.14.97]) 16, 24 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k7MKC9EJ014949 16, 24 -- for {microscopy-at-Microscopy.com} ; Tue, 22 Aug 2006 16:12:09 -0400 16, 24 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 16, 24 -- To: microscopy-at-Microscopy.com 16, 24 -- Date: Tue, 22 Aug 2006 16:14:11 -0400 16, 24 -- MIME-Version: 1.0 16, 24 -- Content-type: text/plain; charset=ISO-8859-1 16, 24 -- Subject: Looking for some video capture recommendations 16, 24 -- Message-ID: {44EB2D53.3411.1471D88D-at-localhost} 16, 24 -- Priority: normal 16, 24 -- X-mailer: Pegasus Mail for Win32 (v3.12c) 16, 24 -- X-Real-ConnectIP: 134.53.14.97 16, 24 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.67 16, 24 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 16, 24 -- Content-Transfer-Encoding: 8bit 16, 24 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k7MKCNVa013909 ==============================End of - Headers==============================
==============================Original Headers============================== 37, 27 -- From keith.morris-at-ucl.ac.uk Tue Sep 5 04:23:01 2006 37, 27 -- Received: from vscani-a.ucl.ac.uk (vscani-a.ucl.ac.uk [144.82.108.29]) 37, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k859N1IH001685 37, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Sep 2006 04:23:01 -0500 37, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 37, 27 -- by vscani-a.ucl.ac.uk with esmtp (Exim 4.60) 37, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 37, 27 -- id 1GKX8y-0001Cc-F2; Tue, 05 Sep 2006 10:22:56 +0100 37, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 37, 27 -- To: {edelmare-at-muohio.edu} 37, 27 -- Cc: {Microscopy-at-microscopy.com} 37, 27 -- Subject: RE: [Microscopy] Looking for some video capture recommendations 37, 27 -- Date: Tue, 5 Sep 2006 10:22:56 +0100 37, 27 -- Message-ID: {001801c6d0cc$df5a4a20$7b865290-at-keithhigrade} 37, 27 -- MIME-Version: 1.0 37, 27 -- Content-Type: text/plain; 37, 27 -- charset="iso-8859-1" 37, 27 -- X-Mailer: Microsoft Office Outlook 11 37, 27 -- Thread-Index: AcbGJ6PBZ9kiWw2FQpaHHhgCHiyF0AAm8PIQ 37, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 37, 27 -- In-Reply-To: {200608222014.k7MKErv0020837-at-ns.microscopy.com} 37, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 37, 27 -- X-UCL-MailScanner: Found to be clean 37, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 37, 27 -- X-Spam-Status: No 37, 27 -- Content-Transfer-Encoding: 8bit 37, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k859N1IH001685 ==============================End of - Headers==============================
==============================Original Headers============================== 27, 23 -- From thomas-moninger-at-uiowa.edu Tue Sep 5 09:18:59 2006 27, 23 -- Received: from hc-smtp1.healthcare.uiowa.edu (hc-smtp1.healthcare.uiowa.edu [129.255.210.32]) 27, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k85EIxtO027457 27, 23 -- for {Microscopy-at-MSA.microscopy.com} ; Tue, 5 Sep 2006 09:18:59 -0500 27, 23 -- Received: from medicine-exch1.medicine.uiowa.edu ([129.255.105.161]) by hc-smtp1.healthcare.uiowa.edu with Microsoft SMTPSVC(6.0.3790.1830); 27, 23 -- Tue, 5 Sep 2006 09:18:58 -0500 27, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 27, 23 -- content-class: urn:content-classes:message 27, 23 -- MIME-Version: 1.0 27, 23 -- Content-Type: text/plain; 27, 23 -- charset="US-ASCII" 27, 23 -- Subject: FW: [Microscopy] RE: Looking for some video capture recommendations 27, 23 -- Date: Tue, 5 Sep 2006 09:18:57 -0500 27, 23 -- Message-ID: {A4AA05CE92DACC43886D133DB94A926E16F36F33-at-medicine-exch1.medicine.uiowa.edu} 27, 23 -- X-MS-Has-Attach: 27, 23 -- X-MS-TNEF-Correlator: 27, 23 -- Thread-Topic: [Microscopy] RE: Looking for some video capture recommendations 27, 23 -- Thread-Index: AcbQzRaxkCt4USNgRM6peHYURhI02wAJ21Mg 27, 23 -- From: "Moninger, Thomas" {thomas-moninger-at-uiowa.edu} 27, 23 -- To: {edelmare-at-muohio.edu} , {Microscopy-at-MSA.microscopy.com} 27, 23 -- X-OriginalArrivalTime: 05 Sep 2006 14:18:58.0818 (UTC) FILETIME=[3A790620:01C6D0F6] 27, 23 -- Content-Transfer-Encoding: 8bit 27, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k85EIxtO027457 ==============================End of - Headers==============================
Yes this RGB filter method works well (in the old B&W tube camera days we used to have filter wheels to move the red, green and blue filter in automatically during image capture). You just put the filter in a suitable place in the light path (i.e. where there's a filter holder) or just rest the filter on the 'light in' point below the condensor (on an inverted system). Capture a red, green & blue B&W 8-bit transmission image and use imageJ or photoshop (in the 'channels' window) to combine the three 8-bit B&W images back into full 24-bit RGB colour. I'll email you a little pdf on how to do it.
The only problem is that you need decent R,G & B filters, and these cost £100 each from Zeiss (not far off a digital camera system). I have the Zeiss filter numbers somewhere if you are interested. I did try a few cheap (sub £20) glass filters from Agar Scientific but filters weren't spectrally good enough for a quality colour picture. Kodak used to make suitable gelatin filters :
But I have never actually tried them (probably it’s the deep blue tri-colour type ones you want).
It also takes a while to swop filters over if you change filters manually (hence the advantage of a filter wheel). Naturally you must not jog anything when collecting the sequence. You can get reasonable colour images resting a compact digital camera against the eyepiece (but its naturally not as good as this RGB filter method with a half decent B&W camera).
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: dainis-at-red5wood.com [mailto:dainis-at-red5wood.com] Sent: 05 September 2006 14:06 To: keith.morris-at-ucl.ac.uk
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Title-Subject: [Filtered] Using RGB filters with monochrome camera
Question: Dear Listers
I intend using a high resolution (Basler) monochrome camera on a Leica DMLM materials microscope with epi illumination. If I take successive images using red, green & blue filters in turn (mounted in the filter magazine between lamp & cube), can I then assemble good quality colour images using something like ImageJ? I notice that there are a few papers on the technique but I'm not clear where it's possible to use the colour filters.
Here is the September 2006 Microscopy Today table of contents. I will close the subscription list for this issue on Friday September 7th, 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 ============================== When Did Agriculture Begin? Stephen W. Carmichael, Mayo Clinic
Microscopy and Microanalysis of Nano-Scale Materials J. R. Michael, L. N. Brewer, D. C. Miller, K. R. Zavadil, S. V. Prasad and P. G. Kotula, Sandia National Lab., Albuquerque, NM
Imaging Gas-Solid Interactions in an Atomic Resolution Environmental TEM Xiao Feng Zhang* and Takeo Kamino,** *Hitachi High Technologies America, Pleasanton, CA, **Hitachi High Technologies Corp., Ibaraki, Japan
Improved Sectioning of Polymers Using an Oscillating Diamond Knife for Transmission Electron Microscopy J.D. Harris* and J.S. Vastenhout,** * The Dow Chemical Company, Midland, MI, ** Dow Benelux B.V., Terneuzen, Netherlands
Applications for Automated Particle Analysis Robert Anderhalt and Lara Swenson, EDAX Inc., Mahwah, NJ
Electromagnetic Simulation Optimizes Design of a Sub-20 nm Resolution Optical Microscope Erik J. Sanchez, Portland State University, Portland, OR
JECP—a Java Electron Crystallography Project X.Z. Li, University of Nebraska, Lincoln, NE
Preparation of the Yeast Pichia pastoris for Transmission Electron Microscopy Benjamin A. Yount, Joan Lin-Cereghino, Geoff P. Lin-Cereghino, and Marcia M. Fox, U. of the Pacific, Stockton, CA
Fly Microdroplets Viewed Big: a Cryo-SEM Approach Stanislav N. Gorb, Max Planck Institute for Metals Research, Stuttgart, Germany
Quantification of Virus Suspensions by Direct Particle Counting Paul R. Hazelton, University of Manitoba, Winnipeg, Canada
Olympus E330 DSLR for Photomicrography with Older Design Microscopes Theodore M. Clarke, Metallurgical Failure Analysis Consultant
Multi-Axial Stage for a Stereo Dissecting Microscope Zhaojie Zhang, University of Wyoming
A Simplified Method for Formulation of Epoxy Resin Embedding Media E. Ann Ellis, Texas A&M University, College Station, TX
A Simple Cleaning Method for Penning Gauges Valery Ray, PBS&T, MEO Engineering Co., Methuen, MA
Negative Stains/Staining 2.5 mM Phosphotungstic Acid, 25µg/mL Bacitracin, pH 7.0 Paul R. Hazelton, University of Manitoba, Winnipeg, Canada
Agar Diffusion Paul R. Hazelton, University of Manitoba, Winnipeg, Canada
New and Interesting at M&M-2007
Industry News
NetNotes IMMUNOCYTOCHEMISTRY – Colloidal gold IMMUNOCYTOCHEMISTRY - Hydrofluoric acid and LR White SAMPLE PREPARATION - Flat embedding SAMPLE PREPARATION - glow discharge SAMPLE PREPARATION - microwave processing SPECIMEN PREPARATION – high vacuum issues with carbon evaporator TEM - Image diffraction pattern rotation TEM - Darkfield imaging TEM - Wehnelt assembly TEM – Powder sample preparation SEM - Critical point drying insects SEM - Observation of polymers morphology SEM - Electromagnetic fields SEM - Critical Point Drier solvents MICROANALYSIS - X-ray emission table
A tip of the hat and my thanks to everyone who contacted me or contributed to the discussion on pixels and image analysis.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333
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==============================Original Headers============================== 7, 18 -- From frank.karl-at-degussa.com Tue Sep 5 14:45:41 2006 7, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k85JjeEj019684 7, 18 -- for {microscopy-at-msa.microscopy.com} ; Tue, 5 Sep 2006 14:45:40 -0500 7, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 7, 18 -- by framailout1.rz.itson.com (8.13.3/8.13.3/Debian-6) with SMTP id k85Jj7cm032637 7, 18 -- for {microscopy-at-msa.microscopy.com} ; Tue, 5 Sep 2006 21:45:38 +0200 7, 18 -- In-Reply-To: {200609011531.k81FV0ws026025-at-ns.microscopy.com} 7, 18 -- Subject: Thanks for responce on: pixels, pixels, little pixels.... 7, 18 -- To: microscopy-at-msa.microscopy.com 7, 18 -- X-Mailer: Lotus Notes Release 6.5.2 June 01, 2004 7, 18 -- Message-ID: {OF61762318.FF416C9A-ON852571E0.006C4A40-852571E0.006C8407-at-degussa.com} 7, 18 -- From: frank.karl-at-degussa.com 7, 18 -- Date: Tue, 5 Sep 2006 15:45:16 -0400 7, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 7, 18 -- 09/05/2006 02:45:39 PM 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
We have been given a small mussel lava preserved in 70% ethanol and salt. Not sure about the salt, maybe from seawater, maybe part of a molecular protocol these researchers are doing.
They want to see it in the SEM. At first I was thinking just run it back to water and do a conventional fix/dehyd/CPD and coat. But then someone asked me about OsO4 in ethanol and skipping the rehydrating steps.
So any ideas? Just do the Os in the 70% or do something different. Not sure what to do about the salt, maybe we can get rid of that another way.
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============================== 8, 21 -- From jmkrupp-at-ucsc.edu Wed Sep 6 13:11:22 2006 8, 21 -- Received: from smtp-prod-mx2.ucsc.edu (smtp-prod-mx2.ucsc.edu [128.114.125.44]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k86IBLZU018209 8, 21 -- for {microscopy-at-microscopy.com} ; Wed, 6 Sep 2006 13:11:21 -0500 8, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 8, 21 -- by smtp-prod-mx2.ucsc.edu (8.13.7/8.13.7) with ESMTP id k86IBGk0027997 8, 21 -- for {microscopy-at-microscopy.com} ; Wed, 6 Sep 2006 11:11:16 -0700 (PDT) 8, 21 -- Received: from [128.114.25.186] (account jmkrupp-at-ucsc.edu [128.114.25.186] verified) 8, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 8, 21 -- with ESMTPA id 69068190 for microscopy-at-microscopy.com; Wed, 06 Sep 2006 11:11:10 -0700 8, 21 -- Mime-Version: 1.0 8, 21 -- Message-Id: {p06230901c124be5afb93-at-[128.114.25.186]} 8, 21 -- Date: Wed, 6 Sep 2006 11:11:13 -0700 8, 21 -- To: microscopy-at-microscopy.com 8, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 8, 21 -- Subject: SEM prep from 70% EtOH 8, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 8, 21 -- X-UCSC-CATS-Information: This message was scanned by the ITS MailScanner 8, 21 -- X-UCSC-CATS-MailScanner: Found to be clean 8, 21 -- X-UCSC-CATS-MailScanner-SpamCheck: 8, 21 -- X-UCSC-CATS-MailScanner-From: jmkrupp-at-ucsc.edu ==============================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 antropos-at-misis.ru as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: antropos-at-misis.ru Name: Ilya Kushneryov
Organization: Moscow Institute of Steel and Alloys
Title-Subject: [Filtered] Where can buy a high temperature microscope?
Question: Hello! Please give me an advice corresponding with Subject! Our Institute need to improve a labor base in part of high temperature electronic microscopy. It is important for us varios high temperature processes to investigate: cristallisation of steel, alloys, non-ferrous metalls, formation of non-metallic inclusions, segregation and others. Where can we find some technical information? We need a list of firms or producer, which can us a such microscope to sell.
Thanks in advance, Ilya Kushneryov Moscow Institute of Steel and Alloys Chair of steel metallurgy
I have a Oxford made cold/hot stage installed in LEO SEM. It is working really good from +100C to -100C. I guess you only need a hot stage that could reach much higher temperature. Gatan has taken over the cold/hot stage business from OXford and the temperature could reach +1250¡ãC. The website is below: http://www.gatan.com/sem/heating_stages.html
Wish this helps. Xiang
----- Original Message ----- X-from: {antropos-at-misis.ru} To: {xyang-at-SMU.CA} Sent: Wednesday, September 06, 2006 8:13 PM
Dear All,
I hope someone can help me out here. I read that alcohol will dissolve the argentaffin granules. How does the chemistry work here? Is it only because of beta-carboline formation or are there other reactions taking place as well? Is the disappearance of argentaffin granules under alcohol fixation confirmatory of its presence if compared to a parallel formaldehyde staining showing its presence?
Also, if I use alcohol fixation, the cells tend to shrink and shrivel. Is there any way I can preserve the morphology with alcohol fixation (ie any other components I can add to alcohol)?
Thanks in advance.
Randy
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We are hiring an Imaging Specialist to work especially on two confocal microscopes, the new one-of-a-kind multiphoton/confocal microscope, calcium imaging and electrophysiological instrumentation. The James C Hogg iCAPTURE Centre of the University of British Columbia is located downtown of Vancouver, in the beautiful province of British Columbia in Canada. Please see the job posting below for more details.
Job Title: Core 3 Imaging Technical Specialist Division: The iCAPTURE Centre, UBC Salary: $48,425 - 51,944
Job summary The primary purpose of this position is to coordinate the iCAPTURE Centre Imaging CORE. This includes supporting all of the iCAPTURE investigators, staff, and trainees in their imaging and biophysical needs using a wide variety of instruments, as well as any external users of the Core. This support includes research and development of new methods, experimental design, sample preparation, instrument optimization for acquisition and analysis, data interpretation, and training of all personnel.
Work performed * Proficient use of all imaging and biophysical instrumentation located in this specialized laboratory. This instrumentation includes two confocal microscopes, the new one-of-a-kind multiphoton/confocal microscope, calcium imaging and electrophysiological instrumentation, all vessel function equipment including myogenic tone instrumentation or other technology as it becomes available and installed in the Core. * Responsible for day-to-day operation of the facility including scheduling, data acquisition, and analysis, data management, instrument maintenance and troubleshooting and providing general support for scientists using the facility. * Train and supervise all other users. * Provide consultation and direct involvement in diverse research projects including data interpretation, trouble shooting for sample preparation, and experimental design. * Remain current on instrumentation, data acquisition and analysis as well as imaging and vessel function techniques and transfer new technology to the Centre. This is done by attending regional user group meetings, workshops, symposia and networking within the flow community. * Experience with and knowledge of many techniques for sample preparation including multi-color imaging, stem cell enumeration, rare event analysis and functional studies of cells, vessels, or organs. * Create new protocols or programs to recognize any particular cell type in an image in consultation with the researcher * Assist or responsible for running large studies which may include organization with other hospital departments as well as recruiting subjects and troubleshooting problems with sample collection.
* Updates on the progress of the study are presented regularly at research group meetings. * Participation in the preparation of manuscripts, grants, and material presented at International Conferences.
Qualifications University Degree (BSc preferred) in Science Minimum 5 years related experience in a laboratory setting Experience with independently conducting specialized experiments related to imaging, particularly with confocal microscopy Computer experience required Accuracy and attention to detail required Effective oral and written communication, interpersonal, multi-tasking and organizational skills Sense of humor a must
Please send your resume to: Kelly Ceron, Human Resources Manager kceron-at-mrl.ubc.ca Only those applicants short listed will be contacted Deadline for application - Sept 15, 2006
==============================Original Headers============================== 9, 17 -- From FChu-at-mrl.ubc.ca Fri Sep 8 19:23:36 2006 9, 17 -- Received: from mail.mrl.ubc.ca (mrl.ubc.ca [137.82.67.155]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k890Na3m028333 9, 17 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Sep 2006 19:23:36 -0500 9, 17 -- Received: from mrlmail-MTA by mail.mrl.ubc.ca 9, 17 -- with Novell_GroupWise; Fri, 08 Sep 2006 17:23:15 -0700 9, 17 -- Message-Id: {4501A6FC020000BC0000B0EE-at-mail.mrl.ubc.ca} 9, 17 -- X-Mailer: Novell GroupWise Internet Agent 7.0.1 9, 17 -- Date: Fri, 08 Sep 2006 17:23:08 -0700 9, 17 -- From: "Fanny Chu" {FChu-at-mrl.ubc.ca} 9, 17 -- To: {Microscopy-at-microscopy.com} 9, 17 -- Subject: Position: Imaging Specialist - UBC iCAPTURE Centre, Vancouver, 9, 17 -- BC 9, 17 -- Mime-Version: 1.0 9, 17 -- Content-Type: text/plain; charset=ISO-8859-1 9, 17 -- Content-Transfer-Encoding: 8bit 9, 17 -- Content-Disposition: inline ==============================End of - Headers==============================
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Email: llmccorm-at-yahoo.com Name: Lindsey
Organization: Henry Ford Hospital
Title-Subject: [Filtered] Digital imaging and analysis
Question: We are working with the Nikon DXM 1200C scope and camera and I saved images in the "fine" setting but we need to do our analysis in the "quick" mode, is there any way to convert between the settings without losing resolution and iamge size? Any suggestions or adivce would be greatly appreciated.
If you have a photograph of any of these, I would be most grateful. Photos of the microscopes alone would be fantastic and preferable, but any photograph would be great.
Thanks so much for your help,
Mike Ganger Weill Cornell Medical College New York NY
==============================Original Headers============================== 8, 20 -- From mganger-at-optonline.net Sun Sep 10 19:05:56 2006 8, 20 -- Received: from mta10.srv.hcvlny.cv.net (mta10.srv.hcvlny.cv.net [167.206.4.205]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8B05uBE012220 8, 20 -- for {Microscopy-at-microscopy.com} ; Sun, 10 Sep 2006 19:05:56 -0500 8, 20 -- Received: from [127.0.0.1] (ool-43514b04.dyn.optonline.net [67.81.75.4]) 8, 20 -- by mta10.srv.hcvlny.cv.net 8, 20 -- (Sun Java System Messaging Server 6.2-6.01 (built Apr 3 2006)) 8, 20 -- with ESMTP id {0J5E00M5KIVZ0A00-at-mta10.srv.hcvlny.cv.net} for 8, 20 -- Microscopy-at-microscopy.com; Sun, 10 Sep 2006 20:04:48 -0400 (EDT) 8, 20 -- Date: Sun, 10 Sep 2006 20:05:54 -0400 8, 20 -- From: Michael Ganger {mganger-at-optonline.net} 8, 20 -- Subject: Old EM Images 8, 20 -- To: Microscopy List Server {Microscopy-at-microscopy.com} 8, 20 -- Message-id: {4504A862.8090806-at-optonline.net} 8, 20 -- MIME-version: 1.0 8, 20 -- Content-type: text/plain; charset=us-ascii; format=flowed 8, 20 -- Content-transfer-encoding: 7BIT 8, 20 -- X-Accept-Language: en-us, en 8, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 8, 20 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mzeldin-at-richmond.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, September 11, 2006 at 14:43:05 ---------------------------------------------------------------------------
Email: mzeldin-at-richmond.edu Name: marty zeldin
Education: K-8 Grade Grammar School
Location: richmond, VA
Question: I'm interested in buying a "started" microscope for a grammar school grandchild. I'd like to keep the cost below $100. Is that possible? If yes, what are your recommendations -- make, model, supplier?
I have a question about magnification and manipulation of pictures in digital format. I am a little bit confused because until now I used photo films, and when they were printed on paper I knew the magnification, which makes the calculation of the "real" magnification of printed pictures taken in TEM easy to know. But now I am using the Megaview III with Analysis and when I save the picture in picture format (to insert in Word reports) I don't know the real magnification. All pictures are 23.3 cm by 17.48 cm. So if I print without rescaling, there is a factor of approx. 2,5x magnification compared to the mag. given by the microscope (a picture taken at 55 000x, when printed on paper, will result in a magnification 130 000x).
Please help me on this issue. Is there a way to easily calculate the real magnification when printing digital images on paper (other than measuring)?
Stephane
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We get asked this fairly regularly by schools and parents (so we have stock replies). Often professional microscopists aren't the best source of information for cheap microscope's as all our optical systems are always over £10,000, and most lab mainstays like our three confocal microscopes come in at nearer £200,000 each (and users still complain about image quality). I assume you are asking about home use with your own kids/grandkids. I would first check out the fun Digital Blue QX-5 I mentioned below - but you need a Windows computer to run it and it's not got fantastic resolution. Generally kids do get very bored with microscopes very quickly when they run out of things to view and soon get fed up with the poor quality of the images (rather unlike their Playstation PS2s, GameCubes & Nintendo DS Lites).
Cheap microscopes under £100 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 and you may make an expensive mistake. Look for old branded 'laboratory' microscopes e.g. Bausch & Lomb, Prior, Leitz, Reichert, Baker, but probably not Tasco toys. Famous existing brands like Zeiss and Olympus attract a high premium. The sellers are often not microscopists though, and many are sold as collector's items and not for 'scientific' use. Also try any local microscope enthusiast clubs - they aren't as common as the many excellent astronomy [telescope] clubs but they are about and have knowledgeable enthusiasts with an eye for low cost quality systems.
There are suppliers geared up to providing cheaper microscopes for schools, so you can ask around at school's science departments, but expect to pay nearer £500 each for a quality setup (although with those like bottom end Meade [www.meade.com] at around £100 you can see something at low mag (~20x objective i.e. around 180x mag) with a quality stained section.
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 large magnifying glass (not the really small hi-mag cheap folding lens ones, try before you buy) - I have a few excellent ones at home for £1 and a good low mag Osram one that includes an illuminating halogen bulb at £8. In general I would say a good stereo dissecting microscope is a good choice (if not the best) for kids as it's great for viewing living things and enlarges what you can see already - look for 40x rather than 4x though. They are a bit expensive though so you would probably have to buy secondhand (look out for branded ones like Bausch & Lomb, and Prior).
Generally prepared slides can rapidly get very boring for under 14s, but living or unusual things (even hamster fur) always attract an audience. Also try your flatbed film scanner, (not the LiDe types that have a very limited depth of focus, and any from around £60 upwards) that will be good for looking at soft static things: leaves, fruit, nuts, household objects (scan at max resolution and try reflected and 'film' mode). In the UK there are sites like http://www.wedgwood-group.com/microscopes_digital.htm that cater for schools and colleges, providing standard compound and stereo microscopes as well as cheap PC video based microscope solutions where the whole class 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 via ebay, but they tend to be expensive and are easily broken by any age-group. Mounted slides keep well though, so 'vintage' ones even from 50 years ago can still look OK - most schools will have some specimen slides knocking about.
At home and our Primary school (under 11) we use the Digital Blue QX-5 (£70) - it's fun but pretty useless for serious microscope work as it's so low res (but at 640 x 480 better than the old Intel QX-3 it replaced). It puts the image on a PC screen. I have one at home for my kids (boy 10 and girl 12) but it only gets occasional use now the novelty has worn off. See http://micro.magnet.fsu.edu/optics/intelplay/index.html (not updated for the QX-5 but all applies - the site even discusses ways of contrast enhancement etc..). Once on the PC the 640x480 images can be manipulated and pasted etc, and the QX-5 does time-lapse for things like crystal growth. Living plants growing and small animals. The similar but far better built Olympus MIC-D was great but being over £500 it was just too expensive for most schools and is now discontinued - there are other similar budget systems about though.
The macro on a good digital camera (like the image stabilised 5MP Canon S2IS going cheap at £200 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. Plus you can the camera for normal photography when you get bored with microscopy.
By the way, if you get a microscope, 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 for books and even microscopes - I have a detailed review on the Digital Blue QX-5 at www.amazon.co.uk). 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: mzeldin-at-richmond.edu [mailto:mzeldin-at-richmond.edu] Sent: 11 September 2006 23:38 To: keith.morris-at-ucl.ac.uk
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mzeldin-at-richmond.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, September 11, 2006 at 14:43:05 ---------------------------------------------------------------------------
Email: mzeldin-at-richmond.edu Name: marty zeldin
Education: K-8 Grade Grammar School
Location: richmond, VA
Question: I'm interested in buying a "started" microscope for a grammar school grandchild. I'd like to keep the cost below $100. Is that possible? If yes, what are your recommendations -- make, model, supplier?
The magnification should be relative to the original microscope's print (or negative) size, if and only if the Megaview captures 100% of the original field of view. If so, the EM's magnification will be correct if you print at the original print size (e.g., 4x5). Any other print size will scale the magnification linearally (i.e., not by area). The math is simple, but entirely dependent on what field size the Megaview captures (again use a scale that is linear, e.g., the horizontal field percentage). Another method would be to image a magnification standard, but I am not a TEM'er and cannot suggest one.
Once you have the mag figured out calculate a relationship that will provide the size (pixels) of a variety of microbars for your microscope's choice of magnifications, e.g., a spreadsheet.
HTH, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
} -----Original Message----- } From: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: September 12, 2006 6:52 AM } To: michael-at-shaffer.net } Subject: [Microscopy] real magnification } } } } } -------------------------------------------------------------- } -------------- } 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 question about magnification and manipulation of } pictures in digital format. } I am a little bit confused because until now I used photo } films, and when they were printed on paper I knew the } magnification, which makes the calculation of the "real" } magnification of printed pictures taken in TEM easy to know. } But now I am using the Megaview III with Analysis and when I } save the picture in picture format (to insert in Word } reports) I don't know the real magnification. } All pictures are 23.3 cm by 17.48 cm. So if I print without } rescaling, there is a factor of approx. 2,5x magnification } compared to the mag. given by the microscope (a picture taken } at 55 000x, when printed on paper, will result in a } magnification 130 000x). } } Please help me on this issue. Is there a way to easily } calculate the real magnification when printing digital images } on paper (other than measuring)? } } Stephane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection } around http://mail.yahoo.com } } ==============================Original } Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Tue Sep 12 04:19:55 2006 6, } 18 -- Received: from web37410.mail.mud.yahoo.com } (web37410.mail.mud.yahoo.com [209.191.91.142]) } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with SMTP id k8C9JtkA030118 } 6, 18 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep } 2006 04:19:55 -0500 } 6, 18 -- Received: (qmail 93133 invoked by uid 60001); 12 Sep } 2006 09:19:54 -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:Conten } t-Type:Content-Transfer-Encoding; } 6, 18 -- } b=H1LGgxOYjFIEW+npxO+UttTVEX88b5cxc1/2GaEfyCyxU3mVjM0rucj1j3KX } LETWoHeNZ/NjKL69dS96Zh44bBhAL6yUDHZA1HT1K+LzHfzfJJp1t8ule9vWaJ } dLZfvxh75xhQI5i+Ug+WTbMdEg09ZnjsCUpfgZpHMh1utW5x8= ; } 6, 18 -- Message-ID: } {20060912091954.93131.qmail-at-web37410.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by } web37410.mail.mud.yahoo.com via HTTP; Tue, 12 Sep 2006 } 02:19:54 PDT 6, 18 -- Date: Tue, 12 Sep 2006 02:19:54 -0700 } (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 } -- Subject: real magnification 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============================== } } __________ NOD32 1.1751 (20060912) Information __________ } } This message was checked by NOD32 antivirus system. } http://www.eset.com } }
==============================Original Headers============================== 7, 21 -- From michael-at-shaffer.net Tue Sep 12 04:47:36 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 k8C9laWR018612 7, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 04:47:36 -0500 7, 21 -- Received: (qmail 3359 invoked from network); 12 Sep 2006 09:52:08 -0000 7, 21 -- Received: from unknown (HELO rarewolf) (michael-at-shaffer.net-at-205.251.84.119) 7, 21 -- by ws6-3.us4.outblaze.com with SMTP; 12 Sep 2006 09:52:08 -0000 7, 21 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 21 -- To: {nizets2-at-yahoo.com} 7, 21 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 21 -- Subject: RE: [Microscopy] real magnification 7, 21 -- Date: Tue, 12 Sep 2006 07:19:35 -0230 7, 21 -- Message-ID: {005401c6d650$c22bb590$4701a8c0-at-rarewolf} 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: Microsoft Office Outlook 11 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 7, 21 -- Thread-Index: AcbWTUcUdJ0sbWy9RhyyLRSA8p7+lAAAe+HA 7, 21 -- In-Reply-To: {200609120921.k8C9Leks030866-at-ns.microscopy.com} ==============================End of - Headers==============================
The easiest solution to this, if it would work for you, is to use the report function in Analysis. Any images printed can have the magnification displayed and calculated automatically. Another option, since you print your images at the same size each time, would be to print an image from all of your common mags from Analysis and use it as a reference when importing them into Word.
Hope this helps, Steve
Steven Lee Chief Technologist Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 75246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Tuesday, September 12, 2006 4:31 AM To: Lee, Steven
Dear listers,
I have a question about magnification and manipulation of pictures in digital format. I am a little bit confused because until now I used photo films, and when they were printed on paper I knew the magnification, which makes the calculation of the "real" magnification of printed pictures taken in TEM easy to know. But now I am using the Megaview III with Analysis and when I save the picture in picture format (to insert in Word reports) I don't know the real magnification. All pictures are 23.3 cm by 17.48 cm. So if I print without rescaling, there is a factor of approx. 2,5x magnification compared to the mag. given by the microscope (a picture taken at 55 000x, when printed on paper, will result in a magnification 130 000x).
Please help me on this issue. Is there a way to easily calculate the real magnification when printing digital images on paper (other than measuring)?
Stephane
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==============================Original Headers============================== 17, 25 -- From StevenLe-at-BaylorHealth.edu Tue Sep 12 06:46:07 2006 17, 25 -- Received: from BHDAEXIMS02.bhcs.pvt (mailhost2.bhcs.com [4.22.141.163]) 17, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8CBk6nD030221 17, 25 -- for {Microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 06:46:07 -0500 17, 25 -- Received: from BHDAEXFE01.bhcs.pvt ([10.5.3.72]) by BHDAEXIMS02.bhcs.pvt with InterScan Messaging Security Suite; Tue, 12 Sep 2006 06:46:06 -0500 17, 25 -- Received: from BHDAEXCH11.bhcs.pvt ([10.5.3.67]) by BHDAEXFE01.bhcs.pvt with Microsoft SMTPSVC(6.0.3790.211); 17, 25 -- Tue, 12 Sep 2006 06:47:17 -0500 17, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 17, 25 -- Content-class: urn:content-classes:message 17, 25 -- MIME-Version: 1.0 17, 25 -- Content-Type: text/plain; 17, 25 -- charset="US-ASCII" 17, 25 -- Subject: RE: [Microscopy] real magnification 17, 25 -- Date: Tue, 12 Sep 2006 06:46:05 -0500 17, 25 -- Message-ID: {74A8B7A904152141BD5AA2761F5D223401C95BC3-at-BHDAEXCH11.bhcs.pvt} 17, 25 -- In-Reply-To: {200609120931.k8C9VNCA017603-at-ns.microscopy.com} 17, 25 -- X-MS-Has-Attach: 17, 25 -- X-MS-TNEF-Correlator: 17, 25 -- Thread-Topic: [Microscopy] real magnification 17, 25 -- Thread-Index: AcbWTkTb0lISWuE2RuWHSEhWFyMWnQAEYUNQ 17, 25 -- From: "Lee, Steven" {StevenLe-at-BaylorHealth.edu} 17, 25 -- To: {Microscopy-at-microscopy.com} 17, 25 -- X-OriginalArrivalTime: 12 Sep 2006 11:47:17.0216 (UTC) FILETIME=[32630600:01C6D661] 17, 25 -- Content-Transfer-Encoding: 8bit 17, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8CBk6nD030221 ==============================End of - Headers==============================
would it not be simpler in the long run to incorporate a scale bar in all of your originally scanned or captured images? Then when they are manipulated the scale bar will still be accurate. It's also useful for publications and general measurement of detail.
Our TEM has a fixed number of magnification steps and it's a relatively trivial task to keep stock images of the scale bars in a folder on the computer then superimpose them. I scan images from negatives at fixed resolutions and have made up scale bars for the most common scan settings. If the scale bar is 10um, 1um, 01.um then measure it in mms and multiply by 100, 1000, 10000 respectively to give you your magnification.
Malcolm
Malcolm Haswell 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
Dear Stephane,
I think that if the accuracy of your digital micrograph magnification is important, there is no substitute for recording images of a standard certified stage micrometer (usually oriented along the major image dimension) under exactly the same optical settings (e.g. zoom!) as your specimens. From these calibration images you can easily calculate the length of each pixel, or the number of pixels per measurement unit (e.g. pixels per micrometer). I once wrote a small program using these data to create a small "micron bar" as a TIFF file for insertion (pasting) into the appropriate digital images. This has the advantage over some commercial systems in that the bar can be placed wherever you like, so as not to obscure any important specimen details. Best wishes,
Jim
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: 12 September 2006 11:30 To: James Chalcroft
Dear listers,
I have a question about magnification and manipulation of pictures in digital format. I am a little bit confused because until now I used photo films, and when they were printed on paper I knew the magnification, which makes the calculation of the "real" magnification of printed pictures taken in TEM easy to know. But now I am using the Megaview III with Analysis and when I save the picture in picture format (to insert in Word reports) I don't know the real magnification. All pictures are 23.3 cm by 17.48 cm. So if I print without rescaling, there is a factor of approx. 2,5x magnification compared to the mag. given by the microscope (a picture taken at 55 000x, when printed on paper, will result in a magnification 130 000x).
Please help me on this issue. Is there a way to easily calculate the real magnification when printing digital images on paper (other than measuring)?
Stephane
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The correct approach is to use a scale bar on each image, in this way there is no abiguity of the dimensions of a micrograph regardless of how it is printed. Using magnification will just lead to errors when documents are printed or copied. Think of your report just like a paper in a Journal. It has to survive any number of reproduction processes from simple printing, to photocopying, scanning and/or faxing. You have no control over the size of the image once it has left your computer, so an absolute scale bar should be embedded permananetly in your image.
Just as you don't trust the nominal magnification of your microscope & film negative , you will need to calibrate the magnification of your microscope & digital negative.
Calibrating the magnification of your digital image would be done the same way you do a negative. Put in a calibrated object, record the "digital" image at a preset pixel resolution at each magnification setting and then calculate the true scale/magnification. The units simply become the scaling dimension (cm, mm, um, nm, pm)/pixel based upon your instruments indicated magnification setting.
Once you have this done you can put any "scale" bar you want on an image since you have a calibration in units/pixel at each magnification. Remember if your camera allows several different resolutions (1K x1K vs 4Kx4K) you will have to compensate for this in your calibration/pixel.
Some digital image capture systems have an option to interface to the microscope and read an apparent magnfication and from this it can calculate and embedd a scale marker on each image. Beware that you should check such calibrations here too, don't blindly trust marker which the imaging system places on your image, it should be close, but there may be errors.
Nestor Your Friendly Neighborhood SysOp
==============================Original Headers============================== 9, 13 -- From zaluzec-at-microscopy.com Tue Sep 12 08:08:12 2006 9, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 9, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8CD8BSk029744 9, 13 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 08:08:12 -0500 9, 13 -- Mime-Version: 1.0 9, 13 -- Message-Id: {p06110402c12c5cefe4dd-at-[206.69.208.22]} 9, 13 -- In-Reply-To: {200609121223.k8CCN3CM008454-at-ns.microscopy.com} 9, 13 -- References: {200609121223.k8CCN3CM008454-at-ns.microscopy.com} 9, 13 -- Date: Tue, 12 Sep 2006 08:08:10 -0500 9, 13 -- To: microscopy-at-microscopy.com 9, 13 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 9, 13 -- Subject: Re: real magnification - Use scale bars! 9, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
In addition to Nestor's advice, one can add a second help to the indispensable scale bar, in the form of a text file, which has the same name than the image, and which encloses all the parameters about the sample, the microscope, the acquisition conditions, and among these, the original magnification, in form of the original microscope magnification (x?0000, calibrated or uncalibrated, film or CCD size, etc), and of the pixel size from the original numerical image (?nm/pixel). The first indicates the conditions in which the image was taken, the second helps for all image processings. Of coarse, it must be corrected if one perform a rescale of the image !
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
nizets2-at-yahoo.com a écrit : } ---------------------------------------------------------------------------- } 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 question about magnification and manipulation } of pictures in digital format. } I am a little bit confused because until now I used } photo films, and when they were printed on paper I } knew the magnification, which makes the calculation of } the "real" magnification of printed pictures taken in } TEM easy to know. But now I am using the Megaview III } with Analysis and when I save the picture in picture } format (to insert in Word reports) I don't know the } real magnification. } All pictures are 23.3 cm by 17.48 cm. So if I print } without rescaling, there is a factor of approx. 2,5x } magnification compared to the mag. given by the } microscope (a picture taken at 55 000x, when printed } on paper, will result in a magnification 130 000x). } } Please help me on this issue. Is there a way to easily } calculate the real magnification when printing digital } images on paper (other than measuring)? } } Stephane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Tue Sep 12 04:19:55 2006 } 6, 18 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.91.142]) } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k8C9JtkA030118 } 6, 18 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 04:19:55 -0500 } 6, 18 -- Received: (qmail 93133 invoked by uid 60001); 12 Sep 2006 09:19:54 -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=H1LGgxOYjFIEW+npxO+UttTVEX88b5cxc1/2GaEfyCyxU3mVjM0rucj1j3KXLETWoHeNZ/NjKL69dS96Zh44bBhAL6yUDHZA1HT1K+LzHfzfJJp1t8ule9vWaJdLZfvxh75xhQI5i+Ug+WTbMdEg09ZnjsCUpfgZpHMh1utW5x8= ; } 6, 18 -- Message-ID: {20060912091954.93131.qmail-at-web37410.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Tue, 12 Sep 2006 02:19:54 PDT } 6, 18 -- Date: Tue, 12 Sep 2006 02:19:54 -0700 (PDT) } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 6, 18 -- Subject: real magnification } 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============================== 6, 29 -- From jacques.faerber-at-ipcms.u-strasbg.fr Tue Sep 12 09:09:58 2006 6, 29 -- Received: from mailhost.u-strasbg.fr (mailhost.u-strasbg.fr [130.79.200.151]) 6, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8CE9wok008747 6, 29 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 09:09:58 -0500 6, 29 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr [130.79.210.2]) 6, 29 -- by mailhost.u-strasbg.fr (8.13.8/jtpda-5.5pre1) with ESMTP id k8CE9sVV085783 6, 29 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 16:09:54 +0200 (CEST) 6, 29 -- Received: from [130.79.54.3] (odhinn.u-strasbg.fr [130.79.54.3]) 6, 29 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id 0AC5810001DF 6, 29 -- for {Microscopy-at-Microscopy.Com} ; Tue, 12 Sep 2006 16:09:32 +0200 (CEST) 6, 29 -- Message-ID: {4506BFA4.3080109-at-ipcms.u-strasbg.fr} 6, 29 -- Date: Tue, 12 Sep 2006 16:09:40 +0200 6, 29 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} 6, 29 -- User-Agent: Thunderbird 1.5.0.5 (X11/20060728) 6, 29 -- MIME-Version: 1.0 6, 29 -- To: Microscopy-at-microscopy.com 6, 29 -- Subject: Re: [Microscopy] real magnification 6, 29 -- References: {200609120931.k8C9VUnJ018015-at-ns.microscopy.com} 6, 29 -- In-Reply-To: {200609120931.k8C9VUnJ018015-at-ns.microscopy.com} 6, 29 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 6, 29 -- Content-Transfer-Encoding: 8bit 6, 29 -- X-IPCMS-MailScanner: Found to be clean 6, 29 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr 6, 29 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-2.0.2 (mailhost.u-strasbg.fr [130.79.200.151]); Tue, 12 Sep 2006 16:09:54 +0200 (CEST) 6, 29 -- X-Virus-Scanned: ClamAV 0.88.4/1871/Tue Sep 12 15:28:18 2006 on mr1.u-strasbg.fr 6, 29 -- X-Virus-Status: Clean 6, 29 -- X-Spam-Status: No, score=-0.5 required=5.0 tests=AWL autolearn=disabled 6, 29 -- version=3.1.4 6, 29 -- X-Spam-Checker-Version: SpamAssassin 3.1.4 (2006-07-25) on mr1.u-strasbg.fr ==============================End of - Headers==============================
Since the original poster mentioned that he is using our camera and therefore also our software, let me weigh in on the issue.
Everyone who has answered to the original posting is correct. The magnification is in fact dependent on what size the images have that you look at (be it on print or on the viewing screen, and also if you are looking at the whole image, or perhaps only a part of it. In the old days of film, this was typically taken care of by printing the entire negative at a fixed image size. It was then just a single conversion factor that had to be taken into account to measure something.
In the digital age, nobody wants to be limited that way anymore. Users want to zoom in on certain areas, they may want a small image or a large print, and it is very easy to do. Calibration then becomes an issue that one has to deal with.
In our software we implemented it the following way:
1) You go through a calibration routine to calibrate a number of magnification settings of the microscope. This is easily done. You put a calibration sample into the scope, take a picture at a given configuration of the camera (full sensor, partial sensor, etc.), and then point out on the screen what the calibration measurement is. From that the software can calculate the pixel calibration (for example 10 nm/pixel) and from then on the software can accurately keep track of the calibration. You can do that for several magnification settings and the software will inter/extrapolate for other magnification settings. We have thus created a fixed link between magnification and calibration. Note: this does not require the magnification of the microscope to be calibrated, only to be reproducible.
2) Once you have an image acquired, you can enter the magnification (either manually or automatically) and the software automatically calibrates the image.
3) You can now display calibration bars on the image or print, and for a print you can also print the magnification. As one of the posters mentioned, you do that with "report" function. A magnification on the screen is not useful, as the computer does not really know what size monitor you are using, and the magnification depends on that. Also, you can use the software directly to make measurements, and don't need to resort to conversion factors and such.
4) We think that it is best to display a scale bar on the image, as that changes with the image when you manipulate the image. For example, if you print an image from the software, both scale bar and magnification will be correct. If you then somehow copy the image and make it, say, smaller, the magnification will be wrong, but the scale bar is still correct.
5) If you want to get information about the acquisition parameters, open the image in our software and press "Alt-Enter". Click on "channel" and it will show you what magnification the images was acquired at, what the exposure time was, etc. This information is stored in the TIF header and thus stays with the image. Unfortunately there are no public TIF tags that can be used for this, so it will only be visible in our software, unless another software writes an import filter for that.
Stephane, look at the scale bar options and the report functionality to set up the software the way you need it. If you have trouble, please call our support. They can help you.
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: Tuesday, September 12, 2006 03:30 To: Mike Bode
Dear listers,
I have a question about magnification and manipulation of pictures in digital format. I am a little bit confused because until now I used photo films, and when they were printed on paper I knew the magnification, which makes the calculation of the "real" magnification of printed pictures taken in TEM easy to know. But now I am using the Megaview III with Analysis and when I save the picture in picture format (to insert in Word reports) I don't know the real magnification. All pictures are 23.3 cm by 17.48 cm. So if I print without rescaling, there is a factor of approx. 2,5x magnification compared to the mag. given by the microscope (a picture taken at 55 000x, when printed on paper, will result in a magnification 130 000x).
Please help me on this issue. Is there a way to easily calculate the real magnification when printing digital images on paper (other than measuring)?
Stephane
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I agree with J. Faerber. It is important to save all the parameters in a readily accessible file. I suggest that this file should be both easily readible in any text editor and reasonably easily parsed on any computer platform. The 'non-standard' TIF headers in analySIS (mentioned by Mike Bode) and the tag groups in DigitalMicrograph are only really useful in those programs. The two formats that come to mind are the .ini format and XML. Because of the easy readability of '.ini' files and built in or freely available parsers for every platform / software package I use, I write my parameters to an '.ini' file with the same name as the image. I use this to process images in DigitalMicrograh, analySIS, SPIDER, EMAN, and EMAN2.
John Minter
J. Faerber wrote (exerpted):
} In addition to Nestor's advice, one can add a second help to the } indispensable scale bar, in the form of a text file, which has the same } name than the image, and which encloses all the parameters about the } sample, the microscope,
The easiest thing is a scale bar instead of magnification, as if you use a map. No magnification there, and no problem. Scale bar is part of the image. It always provides correct reference, regardless of print or display size.
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: {michael-at-shaffer.net} To: {vitalylazar-at-att.net} Sent: Tuesday, September 12, 2006 5:48 AM
I found that taping a magnifying lens to the front of a digital camera (Sony DSC-P32) and setting the focus fixed (if there is the option) yields good pics. Not quite micro, but certainly as good as many dissecting scopes. Some example images may be found at http://flickr.com/photos/mcammer/tags/macro/ Pictures of things like mold and bugs mostly.
} The macro on a good digital camera (like the image stabilised 5MP Canon S2IS } going cheap at £200 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. Plus you can the camera for normal photography when you get } bored with microscopy.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
==============================Original Headers============================== 6, 31 -- From cammer-at-aecom.yu.edu Tue Sep 12 12:42:29 2006 6, 31 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 6, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8CHgT3t020670 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 12:42:29 -0500 6, 31 -- Received: from draco.aecom.yu.edu (draco.aecom.yu.edu [129.98.1.160]) 6, 31 -- by mailgw.aecom.yu.edu (8.12.11.20060308/8.12.11) with ESMTP id k8CHgP95022743 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 13:42:25 -0400 6, 31 -- Received: from draco.aecom.yu.edu (unknown [127.0.0.1]) 6, 31 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id 0E8D03299D0 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 13:40:35 -0400 (EDT) 6, 31 -- X-AuditID: 816201a0-aeecabb000007fbb-46-4506f112dd17 6, 31 -- Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.100]) 6, 31 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id CA8852E5725 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 13:40:34 -0400 (EDT) 6, 31 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.30.137]) 6, 31 -- by post.aecom.yu.edu (Postfix) with ESMTP id BC56126 6, 31 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 13:42:24 -0400 (EDT) 6, 31 -- Message-Id: {5.2.1.1.2.20060912133705.01205b28-at-mailserver.aecom.yu.edu} 6, 31 -- X-Sender: cammer-at-mailserver.aecom.yu.edu 6, 31 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 6, 31 -- Date: Tue, 12 Sep 2006 13:42:25 -0400 6, 31 -- To: microscopy-at-microscopy.com 6, 31 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 6, 31 -- Subject: Re: [Microscopy] RE: AskAMicroscopist: starter microscope for 6, 31 -- a grammar school 6, 31 -- In-Reply-To: {200609120923.k8C9NgWG003309-at-ns.microscopy.com} 6, 31 -- Mime-Version: 1.0 6, 31 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 6, 31 -- X-Brightmail-Tracker: AAAAAA== 6, 31 -- Content-Transfer-Encoding: 8bit 6, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8CHgT3t020670 ==============================End of - Headers==============================
I would suggest that in the age of digital imaging, 'magnification' is a useless concept. An image has only to be displayed on, for example, a 21" screen instead of a 19" screen and the magnification changes.
As Nestor says, a scale bar is much more useful, since it is always correct (provided it has been calibrated) however the image is displayed. From my time as Technical Editor at Microscopy & Analysis, I remember the continual frustration with many contributors sending in images without scale bars and refering to the 'magnification' in the figure caption. Considering not only the processing the image might have gone through before it reached me and since the author didn't know what size the image would be printed at, any reference to 'magnification' was meaningless.
It is interesting to consider scanning probe microscopes. Since they only ever generated digital images, from the very begining, almost nobody involved in SPM uses the term 'magnification'. Images are generally described in terms of the 'field of view' and, as far as I'm aware, all commercial SPM diplay images with either scale bars or 'field of view' data. -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
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==============================Original Headers============================== 4, 14 -- From larry-at-cymru.freewire.co.uk Tue Sep 12 14:30:54 2006 4, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 4, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8CJUrbh000323 4, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 12 Sep 2006 14:30:54 -0500 4, 14 -- Received: from [217.154.248.65] (th1dc-217-154-248-65.dial.mistral.co.uk [217.154.248.65]) 4, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k8CJUfXS001268 4, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 12 Sep 2006 20:30:49 +0100 4, 14 -- Mime-Version: 1.0 4, 14 -- Message-Id: {p06210201c12cb80b25d8-at-[217.154.248.169]} 4, 14 -- Date: Tue, 12 Sep 2006 20:27:49 +0100 4, 14 -- To: Microscopy-at-MSA.Microscopy.Com 4, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 4, 14 -- Subject: Real Magnification 4, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
The Photometrics Applications Group is seeking creative and experienced candidates for the position of Applications Scientist. Among a variety of activities, the Applications Group provides bio-imaging applications oriented support internally and externally, helps to guide product development, produces and refines technical literature, tracks and communicates technology trends and new product opportunities in bio-imaging and opens/maintains lines of communication with the academic sector.
The successful candidate will serve as an essential part of a close-knit, dynamic team of biological imaging experts for cameras, systems, and related products that Photometrics and sister companies supply to the life science community. Duties include identification and monitoring of technology and applications trends, identification of customer requirements and opportunities for new product development. Interface with sales and other marketing personnel to provide on-going product sales training and customer support and understand customer needs and product acceptance. Act as an industry spokesperson for customers, press, and analysts. Identify, establish and manage strategic partnerships, including relationships with customers, suppliers and industry opinion leaders. Candidates should be capable of global travel up to 25% of the time.
Qualifications include skills and knowledge consistent with that usually obtained through an advanced degree, advanced technical knowledge of biological imaging hardware and/or software, experience with bio-chemical and bio-physical methods with a particular emphasis on imaging technologies, and excellent written and verbal communications skills. An in-depth knowledge and understanding of advanced image processing algorithms such as those used in spectral un-mixing, 3-D spatial deconvolution, fluorescence lifetime analysis, fluorescence correlation spectroscopy, FRAP, optical tomography etc. is a plus. Practical knowledge in one or more areas of classical microscopy and/or laser scanning optics, electrical engineering, live cell imaging, whole-animal imaging and/or high-throughput imaging or high-content screening are also a plus.
Interested persons are encouraged to send thier CV/resume and cover letter to:
Karl Garsha Head Applications Scientist Photometrics 3440 E. Brittania Drive Tucson, AZ 85706 kgarsha-at-roperscientific.com
I’m happy to answer informal questions related to the position as well. Please feel free to contact me offlist.
Best, Karl
-- Karl Garsha Applications Scientist Roper Scientific 3440 E. Brittania Drive Tucson, AZ 85706 Office: 520-547-2704
==============================Original Headers============================== 10, 24 -- From garsha-at-itg.uiuc.edu Tue Sep 12 17:19:38 2006 10, 24 -- Received: from outmail128166.authsmtp.net (outmail128166.authsmtp.net [62.13.128.166]) 10, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8CMJcKE013074 10, 24 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 17:19:38 -0500 10, 24 -- Received: from [144.22.2.171] (66-162-43-45.static.twtelecom.net [66.162.43.45]) 10, 24 -- (authenticated bits=0) 10, 24 -- by squirrel.dmpriest.net.uk (8.13.6/8.13.6/Kp) with ESMTP id k8CMJZ6q040989 10, 24 -- for {microscopy-at-microscopy.com} ; Tue, 12 Sep 2006 23:19:36 +0100 (BST) 10, 24 -- Message-ID: {45073274.3080802-at-itg.uiuc.edu} 10, 24 -- Date: Tue, 12 Sep 2006 15:19:32 -0700 10, 24 -- From: Karl Garsha {garsha-at-itg.uiuc.edu} 10, 24 -- Reply-To: kgarsha-at-roperscientific.com 10, 24 -- Organization: Roper Scientific 10, 24 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 10, 24 -- MIME-Version: 1.0 10, 24 -- To: microscopy-at-microscopy.com 10, 24 -- Subject: commercial posting: **Position Announcement** 10, 24 -- Content-Type: text/plain; charset=windows-1252; format=flowed 10, 24 -- Content-Transfer-Encoding: 8bit 10, 24 -- X-Server-Quench: c6ab36b5-42ac-11db-b770-001185d377ca 10, 24 -- X-AuthRoute: OCdyaAgTClZaRR4B CiosDTNPDxgkOxYK DBMeOw5bK0AOTg9W KldyK1tYKloHTlZB SnhYBAkaUV9uOjY1 dAhRbQNNYElEXgxg U08HRFRMFQNqHxgC GBobTRt3dQBZeDAr bTUfJBpdHUR+dU91 SwBXFG8FKzRmOTIc VhJdagdWIVBXfx4R OE0tU3QNfGUHZ3to QgNoYG86NCNULyFQ TxsGLFsWCV4MBSI9 QR9QVXdtJUoeRjky KBpuA1gaG1sXOUg3 PF09GxBw 10, 24 -- X-Authentic-SMTP: 61633138303639.squirrel.dmpriest.net.uk:199/Kp 10, 24 -- X-Report-SPAM: If SPAM / abuse - report it at: http://www.authsmtp.com/abuse 10, 24 -- X-Virus-Status: No virus detected - but ensure you scan with your own anti-virus system! ==============================End of - Headers==============================
On Sep 12, 2006, at 2:20 AM, nizets2-at-yahoo.com wrote:
} Please help me on this issue. Is there a way to easily } calculate the real magnification when printing digital } images on paper (other than measuring)? } Dear Stephane, I agree with those who suggest using a scale bar--in fact that is required for publication in some journals. To calculate the proper size of the scale bar in pixels, you need to take images of a standard at each mag, and the Mag*I*Cal is the best standard I've used. I have no connection with this standard except as a satisfied user. 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 Sep 12 19:07:12 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 k8D07CVZ024800 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 12 Sep 2006 19:07:12 -0500 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id CA6CD2F0A4 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 12 Sep 2006 17:07:11 -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 88B342EFE4 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 12 Sep 2006 17:07:10 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200609120920.k8C9K9t8030285-at-ns.microscopy.com} 4, 22 -- References: {200609120920.k8C9K9t8030285-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {b10344f47bc42b3adeaccb80bbedf27f-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] real magnification 4, 22 -- Date: Tue, 12 Sep 2006 17:10:29 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (batchelder-at-wi.mit.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, September 12, 2006 at 12:48:19 ---------------------------------------------------------------------------
Organization: Keck Microscope Facility, Whitehead Institute
Education: Graduate College
Location: Cambridge, MA, USA
Question: What effect on immunolabeling will dehydrating of a tissue have? Is it possible to immunolabel a tissue then dehydrate it for embedding in paraffin?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (patrick.fairley-at-csauh.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, September 12, 2006 at 20:38:46 ---------------------------------------------------------------------------
Email: patrick.fairley-at-csauh.com Name: patrick fairley
Organization: ohio university college of medicine
Education: Graduate College
Location: St. John Westshore Hospital, Westlake, OH
Question: I am trying to use/salvage a B&L KHS teaching binocular microscope. The optics are in quite good shape but the fine focus adjustment and the variable illumination don't work. Do you know of any resource for details on repairing the fine focus and a wiring/troubleshooting/replacement parts for the electrical? thank you very much pf
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 lamiller-at-uiuc.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: lamiller-at-uiuc.edu Name: Lou Ann Miller
Organization: UIUC
Title-Subject: [Filtered] Micron Bar Tool -- RE: Real Magnification
Question: Just a side note on a useful web tool.
For those who have their calibrations done at the scope...
And who scan negatives and have scanned in a ruler with the identical specs of the negative scanning, or actually still printing...
Below is a calculator to help you figure the magnification part with micron bars.
http://treefrog.cvm.uiuc.edu/cgi-bin/Microna.pl
For example, you scan in a mm ruler and a 15000x negative, plug in 15000, and the calculator will tell you how many mm on the scanned in ruler to make your micron bar for various micron values. Make the micron bar in a layer on top of the ruler image, then pull the layer over and onto your final image. ***** All dpi, scan area, any size change etc must be identical between the 2 files however.
This does not solve your calibration issues, nor is it helpful with many instruments..... but it does help if your scanning negs or printing prints and to keep a lot of clients happy for not having to re-recalculate how to do this over and over and over.
Lou Ann
~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 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
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Email: rstiger-at-ppg.com Name: Rebecca M. Stiger
Organization: PPG
Title-Subject: [Filtered] Relocating TEM
Question: Hello Microscopists:
We are interested in moving our Phillips CM-12 from one facility to another. Having the vendor assist is, of course, one option, but we're interested in exploring other resources as well. Do you know of companies or consultants that manage taking the instrument down, packing, and bringing it back up?
Please feel free to contact me off-list if you'd like.
Your questions pertains to the so-called "pre-embedding" techniques. You will probably find informations related to your exact problem on the web using these key-words. One major disadvantage though is the lack of accessibility of tissues or cells by antibodies. If your target is extracellular your chances are better, but there may still be steric problems. If your antigen is intracellular you have to permeabilize the cells before reacting with the antibodies. Permeabilization has always an effect on the cell/tissue morphology, you have to take this into account. Another point is the label you use. If your secondary antibody is coupled to an enzyme (as it is usually the case in histology) there is a chance that it won't survive the post-labeling treatment. Actually (IMHO) post-embedding techniques are usually preferred because of these heavy drawbacks.
Now I notice that I did not answer your question :-D Yes it is possible, but please consider the above remarks.
Regards,
Stephane
--- batchelder-at-wi.mit.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 } ---------------------------------------------------------------------------- } } Below is the result of your feedback form } (NJZFM-ultra-55). It was submitted by } (batchelder-at-wi.mit.edu) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Tuesday, September 12, 2006 at 12:48:19 } --------------------------------------------------------------------------- } } Email: batchelder-at-wi.mit.edu } Name: Erika Batchelder } } Organization: Keck Microscope Facility, Whitehead } Institute } } Education: Graduate College } } Location: Cambridge, MA, USA } } Question: What effect on immunolabeling will } dehydrating of a tissue have? Is it possible to } immunolabel a tissue then dehydrate it for embedding } in paraffin? } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 7, 12 -- From zaluzec-at-ultra5.microscopy.com Tue Sep } 12 21:29:37 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 } k8D2TZlP004491 } 7, 12 -- for {microscopy-at-microscopy.com} ; Tue, 12 } Sep 2006 21:29:36 -0500 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } (Unverified) } 7, 12 -- Message-Id: } {p06110400c12d1d710424-at-[206.69.208.22]} } 7, 12 -- Date: Tue, 12 Sep 2006 21:29:34 -0500 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: batchelder-at-wi.mit.edu (by way of } Ask-A-Microscopist) } 7, 12 -- Subject: AskAMicroscopist: immunolabeling } & dehydrating of a tissue } 7, 12 -- Content-Type: text/plain; } charset="us-ascii" } ==============================End of - } Headers============================== }
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Hello, I was wondering if anyone was familiar with using the Carbon-flash attachments to the Emitech and Denton desktop sputter coaters. Are you able to get a nice even film of carbon, as well as you can get from using a vacuum evaporator?
Any preference of Emitech versus Denton? We are looking at the turbo version of both units. 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============================== 3, 24 -- From gvrdolja-at-nature.berkeley.edu Wed Sep 13 07:32:15 2006 3, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU [128.32.253.219]) 3, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8DCWFUH031358 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 07:32:15 -0500 3, 24 -- Received: from localhost (localhost [127.0.0.1]) 3, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 5A2B1D977 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 05:32:15 -0700 (PDT) 3, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) 3, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) (amavisd-new, port 10024) 3, 24 -- with ESMTP id 19220-03 for {microscopy-at-microscopy.com} ; 3, 24 -- Wed, 13 Sep 2006 05:32:12 -0700 (PDT) 3, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) 3, 24 -- id C1C98D978; Wed, 13 Sep 2006 05:32:12 -0700 (PDT) 3, 24 -- Received: from localhost (localhost [127.0.0.1]) 3, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id BB3E4D977 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 05:32:12 -0700 (PDT) 3, 24 -- Date: Wed, 13 Sep 2006 05:32:12 -0700 (PDT) 3, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} 3, 24 -- To: microscopy-at-microscopy.com 3, 24 -- Subject: c-flash attachments for sputter coaters 3, 24 -- Message-ID: {Pine.SOC.4.64.0609101808440.29727-at-nature.Berkeley.EDU} 3, 24 -- MIME-Version: 1.0 3, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 3, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 ==============================End of - Headers==============================
I was wondering what temperature a sample may reach under the electron beam of a TEM. I think that for ediffractometry analysis one have to pull the diffraction aperture out of the way, which means no protection for your sample. But crystals melt, and then they lose their crystallinity!
Lets say I use a LAB6 at 200 kV and on a crystal approx. 500 nm in diameter (usually observed around a MAG of 50kx). I know it all depends on the spot size and I don't know if the spot size is standardized between the different microscopes. Lets say I use the smallest spot size. What temperature could reach the crystal if it was deposited on a formavar-coated copper grid (200 mesh)? On a formvar/carbon copper grid? On a Nickel grid? Would the sample be better protected if we sputter coat carbon over the sample instead of over the formvar film?
I wondered how you TEM specialists who work on minerals and crystals, how you know if your material is still crystalline or armorphous under the conditions of observation?
Stephane
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==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Wed Sep 13 07:33:15 2006 7, 18 -- Received: from web37402.mail.mud.yahoo.com (web37402.mail.mud.yahoo.com [209.191.91.134]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k8DCXEd1001197 7, 18 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 07:33:14 -0500 7, 18 -- Received: (qmail 29417 invoked by uid 60001); 13 Sep 2006 12:33:14 -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=uvLpAw0iV2l5lR9fS9kCtMYV4fjPSE0pFmKm5lrdzweiQ0pOQCUD/xWK++Urum9p3A4OFzsrZ3qDR8/hOLbhDbYet135g5p805K15JSYFEXTwRE7JP9OCpZEDrWesRLvOu6eQRh/As5EPgN4ht0ZEvnL1SfaJ5y+nwDjJBHzgf8= ; 7, 18 -- Message-ID: {20060913123314.29415.qmail-at-web37402.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37402.mail.mud.yahoo.com via HTTP; Wed, 13 Sep 2006 05:33:14 PDT 7, 18 -- Date: Wed, 13 Sep 2006 05:33:14 -0700 (PDT) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: local temperature in TEM 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==============================
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 lon.nelson-at-leica-microsystems.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: lon.nelson-at-leica-microsystems.com Name: Lon Nelson
Question: Patrick, if memory serves, this microscope was originally manufactured by Olympus for Bausch & Lomb. Some time ago, Leica Microsystems, Inc. purchased Bausch & Lomb's stereomicroscope line, but did not purchase rights to their compound microscopes (which would include the KHS). This entire compound microscope line has since been discontinued.
Currently, Reichert, Inc. in Buffalo, NY handles service / repair of discontinued B&L stereomicroscopes and they MIGHT be able to help you with the KHS...no guarantees. A contact phone number there is 716 686-3143.
Additionally, you might try I. Miller Precision Optical Instruments in Philadelphia at 215 925-2285.
As a last resort, Google may lead you to someone with repair parts.
Best regards,
Lon M. Nelson Marketing Manager - Life Sciences Leica Microsystems, Inc.
Email: patrick.fairley-at-csauh.com Name: patrick fairley
Organization: ohio university college of medicine
Education: Graduate College
Location: St. John Westshore Hospital, Westlake, OH
Question: I am trying to use/salvage a B&L KHS teaching binocular microscope. The optics are in quite good shape but the fine focus adjustment and the variable illumination don't work. Do you know of any resource for details on repairing the fine focus and a wiring/troubleshooting/replacement parts for the electrical? thank you very much pf
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 swtkeller-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: swtkeller-at-yahoo.com Name: Sandra Keller
Organization: TA/SICCO
Title-Subject: [Filtered] TEM: Looking for a TEM/HRTEM lab in Dallas Fort Worth metroplex
Question: Hi: I am urgently looking for any lab with a TEM/HRTEM that rents to outside users in the Dallas Fort Worth area. Best Regards, Sandra Keller TA/SICCO
The Department of Molecular, Cellular, and Developmental Biology and the Neuroscience Research Institute are sponsoring an advance course on light microscopy. This 4 ½ day workshop will be offered from November 13 through November 17, 2006 and will consist of lectures and laboratory exercises that will run from 9 am to approximately 5 pm each day. The seminar/workshop will be intensive, enabling participants to develop theoretical and hands-on expertise with light microscopes. Attendees will interact closely with the instructors while using modern research grade microscopes, cameras, and computers. The seminars and laboratories will cover basic optical theory and how it pertains to increasing contrast (signal to noise ratio) in biological samples. Fundamental techniques such as fluorescence, phase contrast, Nomarski Differential Interference Contrast, and darkfield imaging will be taught and attendees will use microscopes equipped with these optical enhancement accessories. In addition, the theory and practice of electronic image acquisition (analog and digital) will be presented and attendees will work with low-light cameras, digital image processing computers, and morphometric programs. There are five research grade microscopes, five electronic imaging cameras, two computer workstations, and one confocal microscope. With a maximum enrollment of 10 students, there will be ample opportunity to work with all of the microscopes and cameras. For those so interested, intensive hands-on instruction and guidance on the confocal microscope will be provided. For a fuller description of the workshop please check the web address below. Enrollment forms can be completed online and this workshop provides an opportunity to have a working-vacation in Santa Barbara, California.
For further information on the course please go to the URL address:
On Sep 13, 2006, at 5:33 AM, nizets2-at-yahoo.com wrote:
} I was wondering what temperature a sample may reach } under the electron beam of a TEM. } I think that for ediffractometry analysis one have to } pull the diffraction aperture out of the way, which } means no protection for your sample. But crystals } melt, and then they lose their crystallinity! } } Lets say I use a LAB6 at 200 kV and on a crystal } approx. 500 nm in diameter (usually observed around a } MAG of 50kx). I know it all depends on the spot size } and I don't know if the spot size is standardized } between the different microscopes. Lets say I use the } smallest spot size. What temperature could reach the } crystal if it was deposited on a formavar-coated } copper grid (200 mesh)? On a formvar/carbon copper } grid? On a Nickel grid? Would the sample be better } protected if we sputter coat carbon over the sample } instead of over the formvar film? } } I wondered how you TEM specialists who work on } minerals and crystals, how you know if your material } is still crystalline or armorphous under the } conditions of observation? } Dear Stephane, The final temperature of the sample can be calculated by balancing the energy input to the sample by the beam against the energy that leaves the sample through heat conduction and radiation. Energy is also removed from the sample by secondary electrons and x-rays, but I think that this can be ignored. The energy deposited is equal to the beam current density times the stopping power times the thickness of the sample times the area of the sample. The stopping powers for various substances have been determined and tabulated, but I don't have that table with me. The amount of heat radiated is equal to the Stephan-Boltzman constant times the surface area of the sample times the 4th power of the temperature (in Kelvins), and the amount of heat conducted is equal to the difference in temperature between the sample and the substrate times the thermal conductivity times the effective area of contact, which is somewhere between the projected area of the sample and the perimeter times a "depth of contact", depending on just how well the heat can be carried away from the sample through the substrate. So if you set the energy in (which is not a function of the temperature) equal to the energy out, you can solve for the temperature at steady state, which will be the maximum temperature of the sample. The diffraction aperture is located below the sample, so the sample is exposed to the same amount of beam whether it is in or out. The best way to protect your sample is to use a small beam that can be turned off when images or diffraction patterns are not being taken; a low dose set-up is designed to do precisely that, so use that feature if you have it. (The reason for using a small beam is so that parts of the sample that are not being examined are not being irradiated.) A LaB6 filament can be operated in tip mode, which greatly reduces the intensity of the beam, so you can try this if low dose operation with the largest spot size number (= smallest beam current) still causes your sample to melt. Insert the selected area aperture and spread the beam to a size just larger than the area of the sample defined by that aperture. Carbon coated formvar is a much better conductor than plain formvar, and it is a small benefit to coat with more carbon after the sample is applied to the grid, but the presence of the extra carbon can add background to either the image or diffraction pattern, so it may not be worthwhile, depending on the nature of the sample. Copper is a better conductor than nickel (but not too much better), so there is no advantage to using a nickel grid. I have successfully obtained ED patterns from a variety of crystals without any problems due to melting, including anthracene, which will sublime in the microscope column at room temperature and was examined in a LN2-cooled stage. You should be successful obtaining ED patterns and images from your sample with the proper beam conditions, and you can tell if the sample is still crystalline from the ED pattern or by the FT of an image. 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 Wed Sep 13 12:30:18 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 k8DHUI2h022149 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 13 Sep 2006 12:30:18 -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 8D1C210A170 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 13 Sep 2006 10:30:17 -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 1339F3586B 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 13 Sep 2006 10:30:16 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200609131233.k8DCXLm7001536-at-ns.microscopy.com} 4, 22 -- References: {200609131233.k8DCXLm7001536-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {55df1a240ebae5caad52b3816e3e2cb9-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] local temperature in TEM 4, 22 -- Date: Wed, 13 Sep 2006 10:33:34 -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==============================
We are looking to relocate our EM Facility (SEM´s, TEM´s, Confocal, LM´s) to elsewhere on campus, but before any remodeling or scope moving occurs we need an environmental survey performed. Any suggestions or recommendations as to service providers for doing this in southwest Ohio?
Please, commercial vendors feel free to contact me directly.
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 10, 25 -- From edelmare-at-muohio.edu Wed Sep 13 12:48:11 2006 10, 25 -- Received: from mulnx11.mcs.muohio.edu (mulnx11.mcs.muohio.edu [134.53.6.66]) 10, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8DHmAKj032592 10, 25 -- for {microscopy-at-Microscopy.com} ; Wed, 13 Sep 2006 12:48:11 -0500 10, 25 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 10, 25 -- by mulnx11.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k8DHm3Q5027940 10, 25 -- for {microscopy-at-Microscopy.com} ; Wed, 13 Sep 2006 13:48:06 -0400 10, 25 -- Received: from [192.168.1.23] ([134.53.14.97]) 10, 25 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k8DHm29F002176 10, 25 -- for {microscopy-at-Microscopy.com} ; Wed, 13 Sep 2006 13:48:02 -0400 10, 25 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 10, 25 -- To: microscopy-at-Microscopy.com 10, 25 -- Date: Wed, 13 Sep 2006 13:47:53 -0400 10, 25 -- MIME-Version: 1.0 10, 25 -- Subject: Looking for site survey providers. 10, 25 -- Message-ID: {45080C09.4874.18FF7F55-at-edelmare.muohio.edu} 10, 25 -- Priority: normal 10, 25 -- X-mailer: Pegasus Mail for Windows (4.41) 10, 25 -- Content-type: text/plain; charset=ISO-8859-1 10, 25 -- Content-description: Mail message body 10, 25 -- X-Real-ConnectIP: 134.53.14.97 10, 25 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.66 10, 25 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 10, 25 -- Content-Transfer-Encoding: 8bit 10, 25 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k8DHmAKj032592 ==============================End of - Headers==============================
You need to look at the schmeatic diagrams of the TEM and where the sample sits relative to the objective and diffraction apertures. These are below the sample and as such do not protect your sample. The only things that can protect your sample from heating is minimizing the current density on your sample or minimizing the amount of energy dumped into your sample through inelastic scattering. Your options for the first are several: 1) spread your beam out, 2) use a smaller spot size, 3) use a smaller condenser aperture, or 4) desaturate the filament a little. For minimizing the heating of your sample, you have several options there too: 1) use a cold stage, 2) use a thinner sample, or 3) use a higher accelerating voltage.
I know that when I used 120 kV on glass samples before I had access to a 200 kV machine, coating them with a thin coating of carbon on both sides did help them from softening in the microscope. So I would say yes, carbon coating helps, but it is not the best solution.
You can tell if your sample changes under the beam. While in diffraction mode, simply move the sample to a previously unexposed area and watch the diffraction pattern. If the pattern goes crystalline from amorphous or vice versa, you have a problem and you should call Houston.
-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: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Wednesday, September 13, 2006 5:36 AM To: Walck-at-SouthBayTech.com
Dear listers,
I was wondering what temperature a sample may reach under the electron beam of a TEM. I think that for ediffractometry analysis one have to pull the diffraction aperture out of the way, which means no protection for your sample. But crystals melt, and then they lose their crystallinity!
Lets say I use a LAB6 at 200 kV and on a crystal approx. 500 nm in diameter (usually observed around a MAG of 50kx). I know it all depends on the spot size and I don't know if the spot size is standardized between the different microscopes. Lets say I use the smallest spot size. What temperature could reach the crystal if it was deposited on a formavar-coated copper grid (200 mesh)? On a formvar/carbon copper grid? On a Nickel grid? Would the sample be better protected if we sputter coat carbon over the sample instead of over the formvar film?
I wondered how you TEM specialists who work on minerals and crystals, how you know if your material is still crystalline or armorphous under the conditions of observation?
Stephane
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==============================Original Headers============================== 5, 18 -- From joe.p.neilly-at-abbott.com Wed Sep 13 14:04:25 2006 5, 18 -- Received: from abtmx2.abbott.com (abtmx2.abbott.com [130.36.44.92]) 5, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8DJ4PhH014594 5, 18 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 14:04:25 -0500 5, 18 -- Received: from abtapn561.northamerica.intra.abbott.com (abtapn561.northamerica.intra.abbott.com [10.236.188.103]) 5, 18 -- by abtmx2.abbott.com (Switch-3.1.8/Switch-3.1.7) with ESMTP id k8DJ4OrK001395 5, 18 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 14:04:25 -0500 (CDT) 5, 18 -- To: microscopy-at-microscopy.com 5, 18 -- Subject: Analytical Microscopist Position Available 5, 18 -- MIME-Version: 1.0 5, 18 -- X-Mailer: Lotus Notes Release 6.5.4 HF972 April 26, 2006 5, 18 -- Message-ID: {OFE5558806.D261AE04-ON862571E8.004C79EF-862571E8.0068C590-at-abbott.com} 5, 18 -- From: Joseph P Neilly {joe.p.neilly-at-abbott.com} 5, 18 -- Date: Wed, 13 Sep 2006 14:03:45 -0500 5, 18 -- X-MIMETrack: Serialize by Router on ABTAPN561/ESVR/ABBOTT(Release 6.5.4FP2|September 12, 2005) at 5, 18 -- 09/13/2006 14:04:18, 5, 18 -- Serialize complete at 09/13/2006 14:04:18 5, 18 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
From sinosteeloffice37-at-rediffmail.com Wed Sep 13 14:40:27 2006 Return-Path: {sinosteeloffice37-at-rediffmail.com} Received: from rediffmail.com (webmail39.rediffmail.com [203.199.83.200] (may be forged)) by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k8DJeQX5032601 for {MicroscopyListserverArchive-at-microscopy.com} ; Wed, 13 Sep 2006 14:40:26 -0500 Received: (qmail 1388 invoked from network); 13 Sep 2006 19:37:38 -0000 Received: from unknown (HELO 9d89402a) (80.88.141.72) by mailserver with SMTP; 13 Sep 2006 19:37:38 -0000 Message-ID: {020c01c6d7af$9c4739c0$2900a8c0-at-9d89402a} Reply-To: "Sinosteel Corporation" {info_sinosteel23-at-yahoo.com.cn}
Some of the answer depends on the characteristics of the coating you need. Why would you lean towards a turbo unit? There are reasons.
I use a Denton Desk II with C fiber attachment. Works great. No turbo. I have used Emitech units before and got no/zero support for them in huge contrast to big support from Denton. My current metal coater is a Denton Desk IV TSC with Au/Pd, Pt, Ir or Pd targets. I asked for and got an Edwards XDS5 dry scroll pump so that the whole metal coating system is without oil. Excellent. This is not such a problem with C coating.
gary g.
At 05:33 AM 9/13/2006, you wrote:
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Could someone tell me what year the Proceedings of M&M changed from fully paper printed, to having a CD attached with most of the abstracts? I know it was sometime between 2000 and 2003...
thanks.
John -- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
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==============================Original Headers============================== 5, 24 -- From johnf-at-geology.wisc.edu Thu Sep 14 14:20:06 2006 5, 24 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 5, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8EJK6LY031510 5, 24 -- for {Microscopy-at-Microscopy.Com} ; Thu, 14 Sep 2006 14:20:06 -0500 5, 24 -- Received: from localhost (localhost [127.0.0.1]) 5, 24 -- by localhost (Postfix) with ESMTP id BE43D20D0C 5, 24 -- for {Microscopy-at-Microscopy.Com} ; Thu, 14 Sep 2006 14:20:05 -0500 (CDT) 5, 24 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 5, 24 -- by localhost (geology.wisc.edu [127.0.0.1]) (amavisd-new, port 10024) 5, 24 -- with ESMTP id 02403-06 for {Microscopy-at-Microscopy.Com} ; 5, 24 -- Thu, 14 Sep 2006 14:19:51 -0500 (CDT) 5, 24 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 5, 24 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 5, 24 -- (No client certificate requested) 5, 24 -- by ice.geology.wisc.edu (Postfix) with ESMTP id B40D320D01 5, 24 -- for {Microscopy-at-Microscopy.Com} ; Thu, 14 Sep 2006 14:19:51 -0500 (CDT) 5, 24 -- Mime-Version: 1.0 5, 24 -- Message-Id: {p06230910c12f5bb8bb0a-at-[144.92.206.57]} 5, 24 -- Date: Thu, 14 Sep 2006 14:20:51 -0500 5, 24 -- To: Microscopy-at-Microscopy.Com 5, 24 -- From: John Fournelle {johnf-at-geology.wisc.edu} 5, 24 -- Subject: Proceedings M&M volumes 5, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 24 -- X-Virus-Scanned: amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
Gordon: I use a Denton Desk II with carbon rod, no turbo. I've also used the Emitech K675 which is one of their 8" wafer tools set up for carbon rod. It has a turbo. Both give good coverage; I usually coat for FIB work so I don't have the same criteria as SEM or TEM work would. As Gary says, it will depend on what want and what you're doing with the coating. Ask your sales reps who in your area has coaters you can demo and then you can see if the coatings are what you want.
By the way, I've had the opposite service experience compared with Gary's. (I'm in Texas and I think Gary's on the west coast.)
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 } ---------------------------------------------------------------------------- } } Some of the answer depends on the characteristics of } the coating you need. Why would you lean towards } a turbo unit? There are reasons. } } I use a Denton Desk II with C fiber attachment. } Works great. No turbo. I have used Emitech units } before and got no/zero support for them in huge } contrast to big support from Denton. My current } metal coater is a Denton Desk IV TSC with Au/Pd, } Pt, Ir or Pd targets. I asked for and got an } Edwards XDS5 dry scroll pump so that the whole } metal coating system is without oil. Excellent. } This is not such a problem with C coating. } } gary g. } } } At 05:33 AM 9/13/2006, 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 } } ---------------------------------------------------------------------------- } } } } Hello, } } I was wondering if anyone was familiar with using the Carbon-flash } } attachments to the Emitech and Denton desktop sputter coaters. Are you } } able to get a nice even film of carbon, as well as you can get from using } } a vacuum evaporator? } } } } Any preference of Emitech versus Denton? We are looking at the turbo } } version of both units. } } 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============================== } } 3, 24 -- From gvrdolja-at-nature.berkeley.edu Wed Sep 13 07:32:15 2006 } } 3, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU } } [128.32.253.219]) } } 3, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k8DCWFUH031358 } } 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 } } 07:32:15 -0500 } } 3, 24 -- Received: from localhost (localhost [127.0.0.1]) } } 3, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 5A2B1D977 } } 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 } } 05:32:15 -0700 (PDT) } } 3, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) } } 3, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) } } (amavisd-new, port 10024) } } 3, 24 -- with ESMTP id 19220-03 for {microscopy-at-microscopy.com} ; } } 3, 24 -- Wed, 13 Sep 2006 05:32:12 -0700 (PDT) } } 3, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) } } 3, 24 -- id C1C98D978; Wed, 13 Sep 2006 05:32:12 -0700 (PDT) } } 3, 24 -- Received: from localhost (localhost [127.0.0.1]) } } 3, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id BB3E4D977 } } 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 } } 05:32:12 -0700 (PDT) } } 3, 24 -- Date: Wed, 13 Sep 2006 05:32:12 -0700 (PDT) } } 3, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} } } 3, 24 -- To: microscopy-at-microscopy.com } } 3, 24 -- Subject: c-flash attachments for sputter coaters } } 3, 24 -- Message-ID: {Pine.SOC.4.64.0609101808440.29727-at-nature.Berkeley.EDU} } } 3, 24 -- MIME-Version: 1.0 } } 3, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed } } 3, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 } } ==============================End of - Headers============================== } } } } } ==============================Original Headers============================== } 9, 20 -- From gary-at-gaugler.com Thu Sep 14 10:42:56 2006 } 9, 20 -- Received: from qsmtp4.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 k8EFgtfK016693 } 9, 20 -- for {microscopy-at-microscopy.com} ; Thu, 14 Sep 2006 10:42:55 -0500 } 9, 20 -- Received: (qmail 10177 invoked from network); 14 Sep 2006 08:40:32 -0700 } 9, 20 -- Received: by simscan 1.1.0 ppid: 10168, pid: 10169, t: 0.3232s } 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 qsmtp4 with SMTP; 14 Sep 2006 08:40:32 -0700 } 9, 20 -- Message-Id: {7.0.1.0.2.20060914083326.0267b3f8-at-gaugler.com} } 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } 9, 20 -- Date: Thu, 14 Sep 2006 08:42:57 -0700 } 9, 20 -- To: gvrdolja-at-nature.berkeley.edu } 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} } 9, 20 -- Subject: Re: [Microscopy] c-flash attachments for sputter coaters } 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} } 9, 20 -- In-Reply-To: {200609131233.k8DCXrJf003576-at-ns.microscopy.com} } 9, 20 -- References: {200609131233.k8DCXrJf003576-at-ns.microscopy.com} } 9, 20 -- Mime-Version: 1.0 } 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } ==============================End of - Headers============================== } }
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==============================Original Headers============================== 5, 24 -- From r-holdford-at-ti.com Thu Sep 14 14:29:08 2006 5, 24 -- Received: from arroyo.ext.ti.com (arroyo.ext.ti.com [192.94.94.40]) 5, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8EJT8Kh009436 5, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 14 Sep 2006 14:29:08 -0500 5, 24 -- Received: from dlep35.itg.ti.com ([157.170.170.118]) 5, 24 -- by arroyo.ext.ti.com (8.13.7/8.13.7) with ESMTP id k8EJT202005810 5, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO); 5, 24 -- Thu, 14 Sep 2006 14:29:07 -0500 5, 24 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 5, 24 -- by dlep35.itg.ti.com (8.13.7/8.13.7) with ESMTP id k8EJT2uU026986; 5, 24 -- Thu, 14 Sep 2006 14:29:02 -0500 (CDT) 5, 24 -- Message-ID: {4509AD7E.2000102-at-ti.com} 5, 24 -- Date: Thu, 14 Sep 2006 14:29:02 -0500 5, 24 -- From: Becky Holdford {r-holdford-at-ti.com} 5, 24 -- Organization: SC Packaging Development -- FA Development 5, 24 -- User-Agent: Thunderbird 1.5.0.5 (Windows/20060719) 5, 24 -- MIME-Version: 1.0 5, 24 -- To: gvrdolja-at-nature.berkeley.edu, 5, 24 -- MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 24 -- Subject: Re: [Microscopy] Re: c-flash attachments for sputter coaters 5, 24 -- References: {200609141543.k8EFhLCS017008-at-ns.microscopy.com} 5, 24 -- In-Reply-To: {200609141543.k8EFhLCS017008-at-ns.microscopy.com} 5, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 24 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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Edge effects are common in secondary electron images of samples having the appropriate topography. However, on a flat sample, do you think there may be something equivalent to edge effects in a back scattered electron image? In this case there would be no topography effects but could areas of concentrated mineral give an enhanced signal due to not just the additional high atomic number atoms but also due to the particle distribution?
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============================== 5, 21 -- From dsherman-at-purdue.edu Thu Sep 14 16:02:05 2006 5, 21 -- Received: from 1061exfe04.adpc.purdue.edu (1061exfe04.adpc.purdue.edu [128.210.63.227]) 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8EL25wR031291 5, 21 -- for {microscopy-at-microscopy.com} ; Thu, 14 Sep 2006 16:02:05 -0500 5, 21 -- Received: from exchange.purdue.edu ([172.21.6.23]) by 1061exfe04.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 5, 21 -- Thu, 14 Sep 2006 17:02:04 -0400 5, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 5, 21 -- Thu, 14 Sep 2006 21:02:04 +0000 5, 21 -- User-Agent: Microsoft-Entourage/11.2.5.060620 5, 21 -- Date: Thu, 14 Sep 2006 17:01:58 -0400 5, 21 -- Subject: BS electrons and edge effects 5, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 5, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 5, 21 -- Message-ID: {C12F3B86.143D8%dsherman-at-purdue.edu} 5, 21 -- Thread-Topic: BS electrons and edge effects 5, 21 -- Thread-Index: AcbYQQQbQseNVEQ0EduvwAARJN08Mg== 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: 14 Sep 2006 21:02:04.0878 (UTC) FILETIME=[0834A6E0:01C6D841] ==============================End of - Headers==============================
Yes. BSE generation is a bulk property of the atoms within the interaction volume, therefore variations in local element distribution will cause variation in BSE. I have seen this for instance in crustacean cuticle, where the edges of pores were brighter in BSE imaging because of an increase in Ca concentration at the pore edges. One way to detect this is to compare compositional and topographic images. Which can be confusing if there is both a topographic feature (like, say, a pore) and a compositional feature (like, say, an increase in Ca concentration at the edge of the pore). Then EDS mapping comes in handy. Not to mention changes in BSE images caused by elements migrating due to beam-specimen interactions ...
Phil P.S. "BSE generation is a bulk property of the atoms within the interaction volume." Makes for a great exam question: how can it be truthful to say that a BSE detector can resolve 0.1 Z (atomic number), if Z is always an integer?
} Edge effects are common in secondary electron images of samples having the } appropriate topography. However, on a flat sample, do you think there may } be something equivalent to edge effects in a back scattered electron image? } In this case there would be no topography effects but could areas of } concentrated mineral give an enhanced signal due to not just the additional } high atomic number atoms but also due to the particle distribution? } } 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 -- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
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Question: University of Connecticut Institute of Materials Science
Position in Scanning Electron Microscopy and Optical Microscopy
The Institute of Materials Science (IMS) at UConn is an interdisciplinary center with the threefold mission of fostering education, research and outreach in all areas of the materials sciences. The Microscopy Laboratory is a user facility which houses the main research microscopes (optical, SEM and TEM) for the IMS. There is an opening in the Laboratory for a Scanning Electron Microscopy / Optical Microscopy specialist. The appointee will be involved in a range of academic and industrial projects, and will assist in the operation of the Laboratory including performing routine maintenance and training/assisting users of the microscopes.
Candidates should hold a higher degree (MS or PhD) in Materials Science or a related discipline and must have extensive hands-on SEM and optical microscopy experience. Experience in maintenance of electron microscopes, use of transmission electron microscopy, microscopy of soft materials and/or microtomy would also be beneficial. This is a fixed-term appointment and is available from October 1st. Screening of the applications will begin immediately and will continue until the post is filled. Applications from under-represented groups, including minorities, women and persons with disabilities are encouraged.
Interested candidates should send a curriculum vitae and the names of at least three referees with postal addresses, telephone numbers and Email addresses to: Prof. Mark Aindow, Institute of Materials Science, University of Connecticut, 97 North Eagleville Road, Unit 3136, Storrs, CT 06269-3136 USA. Email: m.aindow-at-uconn.edu
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Title-Subject: [Filtered] Confocal--Chameleon XR service contract
Question: Dear All,
I'm hoping to poll those of you out there who have a 2p laser, specifically Chameleon XR users...do you recommend the service contract?? Our warranty is expiring in a few months and we're weighing our financial options around this issue. Unfortunately, we have already had to have our laser replaced...so we are wary of being without coverage. But of course, the service contract is so expensive, it's hard to swallow the idea of spending the money and not needing any service for the year as well.
So I'm hoping 2p users out there might be able to tell me their experiences with and without a service contract to help us better make this decision.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (zfr_sn-at-yahoo.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, September 14, 2006 at 13:00:00 ---------------------------------------------------------------------------
Email: zfr_sn-at-yahoo.com Name: Sana Ullah
Organization: Govt. College
Education: Graduate College
Location: Lahore,Pakistan
Question: How to prepare a bulk ceramic sample for TEM analysis?
} Edge effects are common in secondary electron images of } samples having the appropriate topography. However, on a } flat sample, do you think there may be something equivalent } to edge effects in a back scattered electron image? } In this case there would be no topography effects but could } areas of concentrated mineral give an enhanced signal due to } not just the additional high atomic number atoms but also due } to the particle distribution?
This is indeed possible, but you should also be careful that your BEI detector isn't also the problem. Both (or all 4) BSED segments need to be perfectly balanced, and I have often observed that putting the sample too near the BSED (short WD) can also enhance edges. In this same regard, I believe that the scintillator type BSED have a large enough acceptance angle to enhance edges at short working distances.
HTH, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
==============================Original Headers============================== 7, 21 -- From michael-at-shaffer.net Fri Sep 15 04:37:26 2006 7, 21 -- Received: from ws6-2.us4.outblaze.com (ws6-2.us4.outblaze.com [205.158.62.197]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k8F9bPIp028593 7, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 15 Sep 2006 04:37:25 -0500 7, 21 -- Received: (qmail 29802 invoked from network); 15 Sep 2006 09:42:52 -0000 7, 21 -- Received: from unknown (HELO rarewolf) (michael-at-shaffer.net-at-205.251.84.119) 7, 21 -- by ws6-2.us4.outblaze.com with SMTP; 15 Sep 2006 09:42:52 -0000 7, 21 -- From: "michael shaffer" {michael-at-shaffer.net} 7, 21 -- To: {dsherman-at-purdue.edu} 7, 21 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 7, 21 -- Subject: RE: [Microscopy] BS electrons and edge effects 7, 21 -- Date: Fri, 15 Sep 2006 07:09:24 -0230 7, 21 -- Message-ID: {001801c6d8aa$d54cadd0$4701a8c0-at-rarewolf} 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: Microsoft Office Outlook 11 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 7, 21 -- Thread-Index: AcbYQdXqqA12qOlKSXKvYdEWIv6saQAaDT8g 7, 21 -- In-reply-to: {200609142102.k8EL2WqX032029-at-ns.microscopy.com} ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (alvarobq-at-fcien.edu.uy) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, September 15, 2006 at 08:40:57 ---------------------------------------------------------------------------
Email: alvarobq-at-fcien.edu.uy Name: Alvaro D. Olivera
Organization: Science University
Education: Undergraduate College
Location: Montevideo - Uruguay
Question:
I'm TEM technician and advanced undergraduate in Biochemistry. I'm trying negative staining whith a small lipo protein HDL like. I'm using CARBON-FORMVAR COATED Cu GRIDS and PTA 2% and 3%. I can't see it. What do you suggest?
If there is a significant difference in the hardness of phases of a specimen, then "flat" specimen could be not really flat, especially if it is prepared by polishing. In cases like this I did observed some "edge effect" in BSE, which is due to the curvature of the edges of phase boundaries.
Also I have observed strong edge effect on bone specimens embedded in a resin (as for TEM) and cut with diamond knife.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 371 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: dsherman-at-purdue.edu [mailto:dsherman-at-purdue.edu] } Sent: Thursday, September 14, 2006 4:03 PM } To: Dusevich, Vladimir } Subject: [Microscopy] BS electrons and edge effects } } } } } -------------------------------------------------------------- } -------------- } 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 } -------------------------------------------------------------- } -------------- } } Edge effects are common in secondary electron images of } samples having the appropriate topography. However, on a } flat sample, do you think there may be something equivalent } to edge effects in a back scattered electron image? } In this case there would be no topography effects but could } areas of concentrated mineral give an enhanced signal due to } not just the additional high atomic number atoms but also due } to the particle distribution? } } 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============================== } 5, 21 -- From dsherman-at-purdue.edu Thu Sep 14 16:02:05 2006 5, } 21 -- Received: from 1061exfe04.adpc.purdue.edu } (1061exfe04.adpc.purdue.edu [128.210.63.227]) } 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with ESMTP id k8EL25wR031291 } 5, 21 -- for {microscopy-at-microscopy.com} ; Thu, 14 Sep } 2006 16:02:05 -0500 } 5, 21 -- Received: from exchange.purdue.edu ([172.21.6.23]) } by 1061exfe04.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); } 5, 21 -- Thu, 14 Sep 2006 17:02:04 -0400 } 5, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by } EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End } Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft } Exchange Server HTTP-DAV ; 5, 21 -- Thu, 14 Sep 2006 } 21:02:04 +0000 5, 21 -- User-Agent: } Microsoft-Entourage/11.2.5.060620 5, 21 -- Date: Thu, 14 Sep } 2006 17:01:58 -0400 5, 21 -- Subject: BS electrons and edge } effects 5, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 5, } 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} } 5, 21 -- Message-ID: {C12F3B86.143D8%dsherman-at-purdue.edu} } 5, 21 -- Thread-Topic: BS electrons and edge effects 5, 21 -- } Thread-Index: AcbYQQQbQseNVEQ0EduvwAARJN08Mg== 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: 14 Sep 2006 21:02:04.0878 (UTC) } FILETIME=[0834A6E0:01C6D841] } ==============================End of - } Headers============================== }
==============================Original Headers============================== 9, 24 -- From DusevichV-at-umkc.edu Fri Sep 15 10:31:25 2006 9, 24 -- Received: from KC-MSXPROTO2.kc.umkc.edu (imap4.exchange.umkc.edu [134.193.143.155]) 9, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8FFVPkT021732 9, 24 -- for {microscopy-at-msa.microscopy.com} ; Fri, 15 Sep 2006 10:31:25 -0500 9, 24 -- Received: from KC-MSX1.kc.umkc.edu ([134.193.32.11]) by KC-MSXPROTO2.kc.umkc.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 24 -- Fri, 15 Sep 2006 10:31:25 -0500 9, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 24 -- Content-class: urn:content-classes:message 9, 24 -- MIME-Version: 1.0 9, 24 -- Content-Type: text/plain; 9, 24 -- charset="us-ascii" 9, 24 -- Subject: RE: [Microscopy] BS electrons and edge effects 9, 24 -- Date: Fri, 15 Sep 2006 10:31:24 -0500 9, 24 -- Message-ID: {032EC4F75A527A4FA58C5B1B5DECFBB33ADC4A-at-KC-MSX1.kc.umkc.edu} 9, 24 -- In-Reply-To: {200609142102.k8EL2hLp032421-at-ns.microscopy.com} 9, 24 -- X-MS-Has-Attach: 9, 24 -- X-MS-TNEF-Correlator: 9, 24 -- Thread-Topic: [Microscopy] BS electrons and edge effects 9, 24 -- Thread-Index: AcbYQxhxuEO00bXyQ022zXZ6m5ESIgAmGVew 9, 24 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} 9, 24 -- To: {microscopy-at-msa.microscopy.com} 9, 24 -- X-OriginalArrivalTime: 15 Sep 2006 15:31:25.0202 (UTC) FILETIME=[01401720:01C6D8DC] 9, 24 -- Content-Transfer-Encoding: 8bit 9, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8FFVPkT021732 ==============================End of - Headers==============================
Dear List Members, } } Let us start by saying that we truly hope this email isn't perceived } as us using this medium as a vehicle for promotion. We also wish to } state that this is in no way an admonishment of Gary for his comments. } He is a valuable contributor to the List and we are in no way } disputing the validity of his claim. However, his innocent comment } about Emitech support has the potential to inflict significant damage } to our businesses, thus we are compelled to respond. } The circumstances have recently changed relative to the ownership of } Emitech as well as their representation here in the US. In March 2005, } Quorum Technologies purchased the ongoing business and assets of } Emitech Ltd. Quorum is the UK manufacturer of Polaron, and both } businesses are located in Southern England. Quorum/Polaron is and } always has been a very Customer Service oriented Company. Since the } acquisition, they have worked very hard and invested heavily in the } Emitech Business, both in the product development and Customer facing } areas. Energy Beam Sciences has been the US Distributor of the } Quorum/Polaron product line for many years and in April 2006, were } appointed the US master distributor for Emitech products as well. This } happened as a result of their commitment to Customer Service as well } as their Customer support capabilities. In January 2005, EBSciences } was purchased by a group of its employees. Since that acquisition, } They also have dedicated much effort and significant resources to } making Customer Service a top priority. EBS has a dedicated Service } Department with top notch technicians and in-house engineering support } to diagnose and repair any problem at their Connecticut facility, or } on-site for larger systems like Cryo-SEM stages. For Applications and } Sales, EBS has a Product Manager who is dedicated exclusively to the } needs of our mutual EM Customers. } } We at Quorum and EBS are united in our desire to serve our Customers } and in our determination to become their first choice for sample } preparation equipment. In fact, we offer any Customer that purchases } or has purchased an instrument from us free telephone/email support } for as long as they own that instument. While we cannot atone for the } sins of the past, we can assure our Customers.of our commitment to } providing the very best in service and support going forward. Sincerely,
Bob Kenhard Manging Director Quorum Technologies
Mike Nesta Managing Director Energy Beam Sciences
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 } ---------------------------------------------------------------------------- } } Some of the answer depends on the characteristics of } the coating you need. Why would you lean towards } a turbo unit? There are reasons. } } I use a Denton Desk II with C fiber attachment. } Works great. No turbo. I have used Emitech units } before and got no/zero support for them in huge } contrast to big support from Denton. My current } metal coater is a Denton Desk IV TSC with Au/Pd, } Pt, Ir or Pd targets. I asked for and got an } Edwards XDS5 dry scroll pump so that the whole } metal coating system is without oil. Excellent. } This is not such a problem with C coating. } } gary g. } } } At 05:33 AM 9/13/2006, 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 } } ---------------------------------------------------------------------------- } } } } Hello, } } I was wondering if anyone was familiar with using the Carbon-flash } } attachments to the Emitech and Denton desktop sputter coaters. Are you } } able to get a nice even film of carbon, as well as you can get from using } } a vacuum evaporator? } } } } Any preference of Emitech versus Denton? We are looking at the turbo } } version of both units. } } 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============================== } } 3, 24 -- From gvrdolja-at-nature.berkeley.edu Wed Sep 13 07:32:15 2006 } } 3, 24 -- Received: from nature.Berkeley.EDU (nature.Berkeley.EDU } } [128.32.253.219]) } } 3, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k8DCWFUH031358 } } 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 } } 07:32:15 -0500 } } 3, 24 -- Received: from localhost (localhost [127.0.0.1]) } } 3, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id 5A2B1D977 } } 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 } } 05:32:15 -0700 (PDT) } } 3, 24 -- Received: from nature.Berkeley.EDU ([127.0.0.1]) } } 3, 24 -- by localhost (nature.berkeley.edu [127.0.0.1]) } } (amavisd-new, port 10024) } } 3, 24 -- with ESMTP id 19220-03 for {microscopy-at-microscopy.com} ; } } 3, 24 -- Wed, 13 Sep 2006 05:32:12 -0700 (PDT) } } 3, 24 -- Received: by nature.Berkeley.EDU (Postfix, from userid 7458) } } 3, 24 -- id C1C98D978; Wed, 13 Sep 2006 05:32:12 -0700 (PDT) } } 3, 24 -- Received: from localhost (localhost [127.0.0.1]) } } 3, 24 -- by nature.Berkeley.EDU (Postfix) with ESMTP id BB3E4D977 } } 3, 24 -- for {microscopy-at-microscopy.com} ; Wed, 13 Sep 2006 } } 05:32:12 -0700 (PDT) } } 3, 24 -- Date: Wed, 13 Sep 2006 05:32:12 -0700 (PDT) } } 3, 24 -- From: Gordon Vrololjak {gvrdolja-at-nature.berkeley.edu} } } 3, 24 -- To: microscopy-at-microscopy.com } } 3, 24 -- Subject: c-flash attachments for sputter coaters } } 3, 24 -- Message-ID: {Pine.SOC.4.64.0609101808440.29727-at-nature.Berkeley.EDU} } } 3, 24 -- MIME-Version: 1.0 } } 3, 24 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed } } 3, 24 -- X-Virus-Scanned: Maia Mailguard 1.0.1 } } ==============================End of - Headers============================== } } } } } ==============================Original Headers============================== } 9, 20 -- From gary-at-gaugler.com Thu Sep 14 10:42:56 2006 } 9, 20 -- Received: from qsmtp4.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 k8EFgtfK016693 } 9, 20 -- for {microscopy-at-microscopy.com} ; Thu, 14 Sep 2006 10:42:55 -0500 } 9, 20 -- Received: (qmail 10177 invoked from network); 14 Sep 2006 08:40:32 -0700 } 9, 20 -- Received: by simscan 1.1.0 ppid: 10168, pid: 10169, t: 0.3232s } 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 qsmtp4 with SMTP; 14 Sep 2006 08:40:32 -0700 } 9, 20 -- Message-Id: {7.0.1.0.2.20060914083326.0267b3f8-at-gaugler.com} } 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } 9, 20 -- Date: Thu, 14 Sep 2006 08:42:57 -0700 } 9, 20 -- To: gvrdolja-at-nature.berkeley.edu } 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} } 9, 20 -- Subject: Re: [Microscopy] c-flash attachments for sputter coaters } 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} } 9, 20 -- In-Reply-To: {200609131233.k8DCXrJf003576-at-ns.microscopy.com} } 9, 20 -- References: {200609131233.k8DCXrJf003576-at-ns.microscopy.com} } 9, 20 -- Mime-Version: 1.0 } 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } ==============================End of - Headers============================== }
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Personally, I would regard an 'edge effect' as a change in contrast due to sample topography which I don't think occurs in BSE imaging.
However, you can get, in many types of sample, 'diffusion' of elements towards morphological and topographical features - such as edges, grain boundaries, etc. Such segregation is 'real' and is revealed by BSE. The 'edge effects' observed in SE imaging are purely a consequence of sample topography on the physics of the imaging method.
As has already been mentioned, preparing a truly flat sample, is difficult. In this case, SE imaging can reveal differences in sample height but BSE imaging will tend to indicate compositional variations.
You should also keep in mind channeling effects, arising from sample crystallography, which give rise to constrast variations unrelated to to composition or topography. And while these are generally 'bulk', that is the whole grain has a contrast determined by orientation and crystallography, it is possible for crystal orientation to be distroted at grain boundaries, leading to constrast changes which could be interpreted as elemental segregation. To separate such effect, you need BSE images plus EDS mapping. -- Larry Stoter
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==============================Original Headers============================== 6, 16 -- From larry-at-cymru.freewire.co.uk Fri Sep 15 14:55:40 2006 6, 16 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 6, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8FJte5S014690 6, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 15 Sep 2006 14:55:40 -0500 6, 16 -- Received: from [217.154.248.230] (th1dc-217-154-248-230.dial.mistral.co.uk [217.154.248.230]) 6, 16 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k8FJt169021192; 6, 16 -- Fri, 15 Sep 2006 20:55:03 +0100 6, 16 -- Mime-Version: 1.0 6, 16 -- Message-Id: {p06210200c130b151023d-at-[217.154.249.234]} 6, 16 -- In-Reply-To: {200609142127.k8ELRoOX011602-at-ns.microscopy.com} 6, 16 -- References: {200609142127.k8ELRoOX011602-at-ns.microscopy.com} 6, 16 -- Date: Fri, 15 Sep 2006 20:53:29 +0100 6, 16 -- To: oshel1pe-at-cmich.edu, Microscopy-at-MSA.Microscopy.Com 6, 16 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 6, 16 -- Subject: [Microscopy] Re: BS electrons and edge effects 6, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
I'm at a quandary to respond to your msg. I see your basis but disagree with it.
The original post was specifically about Emitech and Denton.
} } Hello, } } I was wondering if anyone was familiar with using the Carbon-flash } } attachments to the Emitech and Denton desktop sputter coaters. Are you } } able to get a nice even film of carbon, as well as you can get from using } } a vacuum evaporator? } } } } Any preference of Emitech versus Denton? We are looking at the turbo } } version of both units. } } Thanks.
I have experience with both brands. I think that he is asking for and deserves first-hand feedback about purchase options. But I need more application info before making a more pointed suggestion.
Gordon's question did not, IMO include enough detailed info to make a definitive answer. Thus, all that I could do was to say what I know first-hand at this time. I think that I did so.
I have had pleasure with Denton and lack of pleasure with Emitech. This is based on real experience with both brands. Now, based on my posting, I find that EBS is handling the Emitech line. Having dealt with Mike Dufrane from EBS, this is a very positive action. I think that w/o my posting, many of us would still be in the dark about this new engagement. Blah, blah, EMS is a good supplier...disclaimers, blah, blah.
I'm totally in favor of EBS going in competition with Denton and others. EBS has treated me very well. No complaints. And now I/we know that they represent Emitech. Great. Denton has done great. Thus, no easy binary answer.
The fact is that these units represent a significant monetary investment, like other SEM aspects and begs, if not requires, users to provide positive and negative feedback about products to those organizations and individuals in the process of purchasing capital equipment. Do you advocate that users make purchase decisions based on vague data? I presume not. Thus, you should be able to put my reply in context.
If this reply is inappropriate for the list, I'm sure that Nestor will kill it. Fine. Nevertheless, you have a direct reply.
gary g.
At 09:47 AM 9/14/2006, you wrote: } Gary: } } Your response does not seem to answer the question that Gordon } asked. It seems you have used the question to express your great } pleasure with Denton and your displeasure with Emitech. I do not } think this is the proper forum for such comments. If you have } personal opinions about a particular vendor, it would be more } appropriate to make them off line to someone who asks. I have no } financial interest in either Denton or Emitech. } } David } } {mailto:gary-at-gaugler.com} gary-at-gaugler.com wrote: } } } } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both durainel-at-bsci.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: durainel-at-bsci.com Name: Lita DuRaine
Organization: Boston Scientific
Title-Subject: [Filtered] Gordon / Emitech Sputter Coaters
Question: This is addressed to Gordon Ante Vrdoljak: We have an Emitech K550X Sputter Coater with the carbon flash attachment and have been very satisfied with the results. The coatings so far have appeared evenly distributed on our samples. Customer service for our location in Fremont, CA has been great. I like the unit because we can sputter various metals as well.
Hope that helps, Lita DuRaine Boston Scientific Manager Microscopy Lab
Dear Alvaro, interesting question....& a } lazy sunday {.....
Normally (e.g. as I use it sometimes for negative staining of virus particles) you will have at least two negative staining solutions: i) hydrous PTA (1, 2 or other percentage) ii) hydrous uranylacetate 1-4%.......and sometimes also you will be informed also about a certain pH your solution should for such preparations....
For (small) lipoproteins (esp. for HDL-like) I am not quite sure about wether } simply saying { so or } giving advice { in that way is helpful.
As I understand negative staing, (a) positive result(s) will also depend on a given pH (and therefore [with WHAT ingredients?] buffering the solution eventually to WHICH "special" pH), and (absorption/adhesion) technique of specimen preparation. I'm not an expert on that field, but i don't think your carbon-coated formvar-filmed copper grids are the biggest problem....but,. if I remember correctly, there are several (also older)papers/articles on negative staining techniques for e.g. liposomes (for an imagination on such, try the search phrase } } "negative staining","liporotein" { {) on google, resulting in a lot of papers and statements about some very specific caudeles for adhesion of lipoprotein material....(for instance: see*) below.
Some prefer also prefixation of mixed particles with hydrous Osmium tetroxide before negative staining with the heavy metal solution, sometimes only hydrous, somtetimes with an additive (like sucrose)..... so it may depend also on the type of material you'd like to have demonstrated and wether you are able to get the (macro-)molecule adhering in a stabilized form to the formvar-carbon-surface of the grid or not (cf. *)
Once determined, [by trial&error (perhaps, unluckily)] how to proceed with the grids' properties = hydrophilization and immediate use or storage for months...see *), adhesion time/technique, eventually prestabilization of the specimen by additional fixation, the right negative staining solution (also in terms of i) element [PTA, UO2, AmmMolybdat,Uranylformate, etc.], ii)concentration, iii) pH of the solution, iv) specific additives necessary?)
the technique itself certainly yields reliable and rapid results, even within 5-10 minutes (as I have read in some papers) ==} THAT's what I would like to wish you in overcoming your problem.
I am sure you will get more specific advice via this forum from experts on the matter....
Best regards Wolfgang Muss, Salzburg, AUSTRIA
cf: *) Original paper in: J Lipid Res. 1980 Nov;21(8):981-92. Unilamellar liposomes made with the French pressure cell: a simple preparative and semiquantitative technique. Hamilton RL Jr, Goerke J, Guo LS, Williams MC,Havel RJ. Ex Material & Methods-section: Negative staining of liposomes often causes artifactual images resulting from disruption of small liposomes that subsequently form larger multilamellar structures. Although the mechanism of this process is not understood, it may be caused in part by the intense hydrophobicity of freshly evaporated carbon on grid surfaces. Our attempts to modify the surface film of freshly prepared carbon-coated grids (200-300 mesh copper grids, Fullum, Schenectady,NY) by glow discharge, polylysine, or addition of albumin to sample or grid did not prove satisfactory. However, we have learned empirically that parlodioncovered grids with carbon films made with a Siemens evaporator (VI3 G 500) permit even spreading of liposomes without apparent structural changes, provided that the grids have been aged 6- 12 months on the shelf. Carbon rods for filming were obtained from Ringsdorff-Herke (Spektral Kohlen, Bonn-Bad Godesberg, West Germany). With such “matured” carbon-coated grids, liposomes were prepared for electron microscopy with 2% potassium phosphotungstate at pH 6.45-6.5 by the drop procedure described previously (1 1). We found that the lipid concentration in the sample was also important for preparing uniform spreads. Optimal results were more consistently obtained with samples containing 1- 10 mg/ml PC. Improved results can be obtained for more dilute samples by increasing contact time of the sample on the grid to 2-3 min, and by using a tighter 400 mesh grid.......
308 results with a Google search phrase: } } "negative staining" "of HDL" { { : some of them can be retrieved without charges (esp. older ones out from Journal Lipid Res),
Cf. also (free download) at: http://www.jlr.org/cgi/reprint/41/2/285 : J. Lipid Res. Chung and Dashti, 41 (2): 285. Lipolytic remnants of human VLDL produced in vitro: effect of HDL levels in the lipolysis mixtures on the apoCs to apoE ratio and metabolic properties of VLDL core remnants
or another paper at: http://www.pubmedcentral.gov/picrender.fcgi?artid=292037&blobtype=pdf
also see: http://em-outreach.ucsd.edu/web-course/Sec-III.F/Sec-III.F.html, section c. Contrast enhancement....
NB/PS: Performing an original search, } "negative staining" "of HDL" { in PubMed Central will retrieve 55 citations.
Zitat von alvarobq-at-fcien.edu.uy:
} } } } ---------------------------------------------------------------------------- } 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 (alvarobq-at-fcien.edu.uy) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on } Friday, September 15, 2006 at 08:40:57 } --------------------------------------------------------------------------- } } Email: alvarobq-at-fcien.edu.uy } Name: Alvaro D. Olivera } } Organization: Science University } } Education: Undergraduate College } } Location: Montevideo - Uruguay } } Question: } } I'm TEM technician and advanced undergraduate in Biochemistry. } I'm trying negative staining whith a small lipo protein HDL like. I'm } using CARBON-FORMVAR COATED Cu GRIDS and PTA 2% and 3%. I can't see it. } What do you suggest? } } Many thanks, Alvaro. } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 9, 12 -- From zaluzec-at-ultra5.microscopy.com Fri Sep 15 08:47:52 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 } k8FDlpKs009958 } 9, 12 -- for {microscopy-at-microscopy.com} ; Fri, 15 Sep 2006 08:47:51 -0500 } 9, 12 -- Mime-Version: 1.0 } 9, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) } 9, 12 -- Message-Id: {p06110406c1305f547915-at-[206.69.208.22]} } 9, 12 -- Date: Fri, 15 Sep 2006 08:47:50 -0500 } 9, 12 -- To: microscopy-at-microscopy.com } 9, 12 -- From: alvarobq-at-fcien.edu.uy (by way of Ask-A-Microscopist) } 9, 12 -- Subject: AskAMicroscopist: negative staining whith a small lipo } protein } 9, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== }
==============================Original Headers============================== 24, 37 -- From W.Muss-at-salk.at Sun Sep 17 08:22:57 2006 24, 37 -- Received: from hermes.salk.at (hermes.lks.at [193.170.167.9] (may be forged)) 24, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8HDMuF0000736 24, 37 -- for {microscopy-at-microscopy.com} ; Sun, 17 Sep 2006 08:22:57 -0500 24, 37 -- Received: from localhost (localhost [127.0.0.1]) 24, 37 -- by hermes.salk.at (Postfix) with ESMTP id 3630BC3875; 24, 37 -- Sun, 17 Sep 2006 15:22:55 +0200 (CEST) 24, 37 -- X-Virii-Scanned: Kaspersky Antivirus at salk.at 24, 37 -- Received: from hermes.salk.at ([127.0.0.1]) 24, 37 -- by localhost (n1ex218.lks.local [127.0.0.1]) (amavisd-new, port 10024) 24, 37 -- with ESMTP id 9x3X7vfafMoc; Sun, 17 Sep 2006 15:22:54 +0200 (CEST) 24, 37 -- Received: from hermes.lks.at (hermes.salk.at [192.168.13.210]) 24, 37 -- by hermes.salk.at (Postfix) with ESMTP id AAF05C3874; 24, 37 -- Sun, 17 Sep 2006 15:22:54 +0200 (CEST) 24, 37 -- Received: from localhost (localhost [127.0.0.1]) 24, 37 -- by hermes.lks.at (Postfix) with ESMTP id 74D905A900A; 24, 37 -- Sun, 17 Sep 2006 15:22:54 +0200 (CEST) 24, 37 -- Received: from hermes.lks.at ([127.0.0.1]) 24, 37 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 24, 37 -- with ESMTP id 06221-01; Sun, 17 Sep 2006 15:22:54 +0200 (CEST) 24, 37 -- Received: from salk.at (webmail.salk.at [192.168.13.209]) 24, 37 -- by hermes.lks.at (Postfix) with ESMTP id 03DDF5A901F; 24, 37 -- Sun, 17 Sep 2006 15:22:53 +0200 (CEST) 24, 37 -- Received: from m109p013.adsl.highway.telekom.at (m109p013.adsl.highway.telekom.at [62.47.181.141]) 24, 37 -- by webmail.salk.at (IMP) with HTTP 24, 37 -- for {pawma-at-192.168.13.210} ; Sun, 17 Sep 2006 15:22:53 +0200 24, 37 -- Message-ID: {1158499373.450d4c2de4be4-at-webmail.salk.at} 24, 37 -- Date: Sun, 17 Sep 2006 15:22:53 +0200 24, 37 -- From: Wolfgang Muss {W.Muss-at-salk.at} 24, 37 -- To: alvarobq-at-fcien.edu.uy, microscopy-at-microscopy.com 24, 37 -- Subject: [Microscopy]Re:AskAMicroscopist: negative staining whith a small lipo protein 24, 37 -- MIME-Version: 1.0 24, 37 -- Content-Type: text/plain; charset=ISO-8859-1 24, 37 -- Content-Transfer-Encoding: 8bit 24, 37 -- User-Agent: Internet Messaging Program (IMP) 3.2.8 24, 37 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at 24, 37 -- X-Scanned-By: SALK-Content-Filter ==============================End of - Headers==============================
I am choosing between two Zeiss 5x objectives [Plan-Apochromat (420630-9900); EC Plan-Neofluar (420330-9901)] to be used for both diagnostic pathology and research. Most of the specimens I will be examining are H&E or horse radish peroxidase (diaminobenzidine) stained. The numerical aperatures (0.16) and fields of view (25 mm) are identical for both objectives. Zeiss claims better color correction and flatness of field for the Plan-Apochromat versus the EC Plan-Neofluar.
The Plan-Apochromat has a flatter % transmission/ wavelength curve than the EC Plan-Neofluart and extends into the IR range (80% transmission -at- 1100 nm). The transmission of the EC Plan-Neofluar extends a bit further into the near UV (about 325 nM) than that of the Plan-Apochromat. For some reason, DIC is not listed as a potential application of the Plan-Apochromat, but not the EC Plan-Neofluar.
I'm sure that both are great objectives, but which one would be the better choice for my applications? Also, does anyone have experience using the Zeiss Plan-Apochromat 40X/0.95 Corr (42066-9970) objective?
Thanks in advance for your responses,
Bill Howell, M.D., Ph.D.
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Email: twigg-at-estd.nrl.navy.mil Name: Mark Twigg
Organization: Naval Research Laboratory
Title-Subject: [Filtered] Ghosted Image Window in Digital Micrograph
Question: The Dell Dimension 8400 (running Windows XP) on which I am running Digital Micrograph recently crashed. After the crash the Image widow in Digital Micrograph has been ghosted, so that I cannot insert my Ultrascan CCD camera or attempt to record an image. I have rebooted the PC a number of times, but to no avail. Perhaps the PC-savy people among you have some suggestions for me here. I have mostly used Macs and am therefore not familiar with PC ailments. Also, would it help to restart or reset the First Light Digital Camera controller?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sheila.blouin-at-umb.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, September 17, 2006 at 21:25:08 ---------------------------------------------------------------------------
Email: sheila.blouin-at-umb.edu Name: Sheila
Organization: University of Massachusetts, Boston
Education: Undergraduate College
Location: Boston, MA, USA
Question: What does a microscopist do all day? What kind of education do you have to have?
Dear all, is there anybody out there who has experience with removing a small drop of diffusion pump (non silicone) oil from an EDS light element detector window? Window is made of 4um Be... (SLEW window from Roentec). I tried to remove the oil from the colliminator surface with multiple washing in 100% alcohol and it seems that I succeeded but I don`t know if I should use this "protocoll" on the more delicate Be-window...
We have a LEO 912 Omega TEM and would like to obtain a second-hand double tilt holder (in any condition) for electron diffraction analysis of crystalline specimens. Please contact me should you be able to help.
Thanks in advance.
Mohamed
************************************ Mohamed Jaffer Electron Microscope Unit University of Cape Town Private Bag Rondebosch, 7701 South Africa
Tel: +27 21 6503354 Fax: + 27 21 6891528
Email: mjaffer-at-science.uct.ac.za
************************************
==============================Original Headers============================== 13, 20 -- From mjaffer-at-science.uct.ac.za Mon Sep 18 07:58:37 2006 13, 20 -- Received: from mail.uct.ac.za (mail.uct.ac.za [137.158.128.3]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8ICwaN3009190 13, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 18 Sep 2006 07:58:36 -0500 13, 20 -- Received: from [137.158.37.13] (helo=science.uct.ac.za) 13, 20 -- by mail.uct.ac.za with esmtp (Exim 4.44 (FreeBSD)) 13, 20 -- id 1GPIhk-0000ck-VF 13, 20 -- for Microscopy-at-microscopy.com; Mon, 18 Sep 2006 14:58:33 +0200 13, 20 -- Message-ID: {450E97F9.F8C01F31-at-science.uct.ac.za} 13, 20 -- Date: Mon, 18 Sep 2006 14:58:33 +0200 13, 20 -- From: Mohamed A Jaffer {mjaffer-at-science.uct.ac.za} 13, 20 -- Reply-To: mjaffer-at-science.uct.ac.za 13, 20 -- Organization: University of Cape Town 13, 20 -- X-Mailer: Mozilla 4.75 [en] (Windows NT 5.0; U) 13, 20 -- X-Accept-Language: en 13, 20 -- MIME-Version: 1.0 13, 20 -- To: Microscopy-at-microscopy.com 13, 20 -- Subject: double tilt holder 13, 20 -- Content-Type: text/plain; charset=us-ascii 13, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Microscopist are such a diverse group I suspect it is not possible to give a clear cut answer.
Most of us spend time with a computer/notebook recording our results. Many of us write reports documenting our results and in many cases interpreting how the results dove-tail to provide a consistent, self-logical portrait of our work.
For me, currently, it means isolating materials, sometimes particles, sometime making big things into smaller "microscope-sized " specimens. I typically use Energy Dispersion Spectroscopy (EDS) as well as SEM and TEM examination. Other problems require the polarized light microscope (both for determination of morphology as well as optical crystallography) and microchemical test. These test run from the simple (cold dilute mineral acid makes limestone bubble) to the more complex (test for sulfate with barium chloride and sodium rhodizonate). In a former position it also meant microtoming polymers, testing fillers for asbestos and alpha quartz as well as running the IR microscope and interpreting IR spectra and entry use of the GC/MS (I set-up runs with caned programs and matched results to libraries).
Education? First get a degree in a physical science. While your doing this take courses and avail yourself of options to learn more about microscopy related skills.
In geology you could learn about optical mineralogy, optical crystallography, SEM/EDS and WDS. Biology: pollen, diatoms, "pond water microscopy", TEM, hair, feathers and other "biologicals" in the environment Material sciences: SEM, TEM, AFM, EDS Chemistry: chemistry, reactions, semiquantative test Forensic science: all that "CSI" stuff, but without the glamor
We learn to think logical and self consistently. We use the scientific method to gather and then challenge our data and results.
The list continues. Many of us are self-taught. That is, we study a book or test procedure and worked at it until we understand it. Most of us still take courses, many of us have either a scope at home, or a personal project at work that we're fooling around with. We spend some of our free time reading the journals and literature, both current and older classical works. We develop some amount of computer skill, if not with the ubiquitous computer that seems to run all our modern electron scopes, word, powerpoint and excel as well as imaging software. We learn to compose photomicrographs as to rival art works (I'm still working on this!!).
Can you program? Do you like sales? Most of the great optical salesmen and women I know are excellent light microscopist. How do you feel about field-work?
Life maybe a highway, but microscopy can be a road atlas. Where do you want to go?
stay safe.........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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sheila.blouin-at-umb .edu To: frank.karl-at-degussa.com cc: 09/18/2006 08:29 Subject: [Microscopy] AskAMicroscopist: What does a microscopist do all day? AM Please respond to sheila.blouin
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sheila.blouin-at-umb.edu) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, September 17, 2006 at 21:25:08 ---------------------------------------------------------------------------
Email: sheila.blouin-at-umb.edu Name: Sheila
Organization: University of Massachusetts, Boston
Education: Undergraduate College
Location: Boston, MA, USA
Question: What does a microscopist do all day? What kind of education do you have to have?
I am trying to understand what is happening with a set of BSE images. Your comments will be welcome!
Below are links to two images. The first (1.5 Mb) shows two BSE images of a nickel based super alloy (Ni-Cr-Fe-Ti). Both were acquired using a 4-diode detector, 5 kV. beam, and as close to zero degrees tilt as I could set the stage. The top of the first image is in the "as polished" condition, the lower portion of the image is after a very light electro-etch. Notice the difference in channeling contrast. Z-contrast seems largely unaffected (e.g. Ti and Cr carbide inclusions). Perhaps the difference is from my inability to set EXACTLY the same tilt, but they should be within a few degrees (or better) of the same value.
Why the dramatic reversal of contrast for some grains????
The second image is simply a 60 degree tilt SE image of the same general area to show relief of the carbides due to both polishing and the etch. ...Not much.
I would be willing to bet that an oxide film may have formed during etching and the thickness of the film varies with grain orientation. I would try collecting the BSE images at a higher voltage (20 KV) to minimize the effect of the film.
On 9/18/06 9:49 AM, "NWWhite-at-bwxt.com" {NWWhite-at-bwxt.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 All, } } I am trying to understand what is happening with a set of BSE images. } Your comments will be welcome! } } Below are links to two images. The first (1.5 Mb) shows two BSE images } of a nickel based super alloy (Ni-Cr-Fe-Ti). Both were acquired using a } 4-diode detector, 5 kV. beam, and as close to zero degrees tilt as I } could set the stage. The top of the first image is in the "as polished" } condition, the lower portion of the image is after a very light } electro-etch. Notice the difference in channeling contrast. Z-contrast } seems largely unaffected (e.g. Ti and Cr carbide inclusions). Perhaps } the difference is from my inability to set EXACTLY the same tilt, but } they should be within a few degrees (or better) of the same value. } } Why the dramatic reversal of contrast for some grains???? } } The second image is simply a 60 degree tilt SE image of the same general } area to show relief of the carbides due to both polishing and the etch. } ...Not much. } } } http://www.bwxt.com/operations/images/sem/126867_859.jpg } } } http://www.bwxt.com/operations/images/sem/126866.jpg } } } Thanks, } Woody White } BWXT Services } Lynchburg, VA } } } } } ==============================Original Headers============================== } 14, 26 -- From nwwhite-at-bwxt.com Mon Sep 18 08:48:46 2006 } 14, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) } 14, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k8IDmkt4030869 } 14, 26 -- for {microscopy-at-msa.microscopy.com} ; Mon, 18 Sep 2006 08:48:46 } -0500 } 14, 26 -- Received: from ([131.184.13.224]) } 14, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.2215552; } 14, 26 -- Mon, 18 Sep 2006 09:48:37 -0400 } 14, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by } bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); } 14, 26 -- Mon, 18 Sep 2006 09:48:37 -0400 } 14, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 14, 26 -- Content-class: urn:content-classes:message } 14, 26 -- MIME-Version: 1.0 } 14, 26 -- Content-Type: text/plain; } 14, 26 -- charset="US-ASCII" } 14, 26 -- Subject: BSE image response question } 14, 26 -- Date: Mon, 18 Sep 2006 09:48:37 -0400 } 14, 26 -- Message-ID: } {360DDAABED436B49BA397357CCEE8BB703D063-at-BWXSPO01.BWXS.BWXTECH.NET} } 14, 26 -- X-MS-Has-Attach: } 14, 26 -- X-MS-TNEF-Correlator: } 14, 26 -- Thread-Topic: BSE image response question } 14, 26 -- Thread-Index: AcbbKSRJZ3PRklUEQNiswlKEqWhmhg== } 14, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} } 14, 26 -- To: "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} } 14, 26 -- X-OriginalArrivalTime: 18 Sep 2006 13:48:37.0909 (UTC) } FILETIME=[247F1C50:01C6DB29] } 14, 26 -- Content-Transfer-Encoding: 8bit } 14, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k8IDmkt4030869 } ==============================End of - Headers==============================
==============================Original Headers============================== 8, 23 -- From david.r.hull-at-nasa.gov Mon Sep 18 09:18:08 2006 8, 23 -- Received: from NDJSVWS01.ndc.nasa.gov (ndjsvws01.ndc.nasa.gov [198.120.25.81]) 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8IEI7XC009286 8, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 18 Sep 2006 09:18:07 -0500 8, 23 -- Received: from ndjsxgw01.ndc.nasa.gov ([129.166.32.111]) by NDJSVWS01.ndc.nasa.gov with InterScan Messaging Security Suite; Mon, 18 Sep 2006 09:18:05 -0500 8, 23 -- Received: from NDJSEVS16.ndc.nasa.gov ([129.166.32.127]) by ndjsxgw01.ndc.nasa.gov with Microsoft SMTPSVC(6.0.3790.1830); 8, 23 -- Mon, 18 Sep 2006 09:17:54 -0500 8, 23 -- Received: from 129.166.32.15 ([129.166.32.15]) by NDJSEVS16.ndc.nasa.gov ([129.166.32.136]) via Exchange Front-End Server mail01.ndc.nasa.gov ([129.166.32.105]) with Microsoft Exchange Server HTTP-DAV ; 8, 23 -- Mon, 18 Sep 2006 14:17:54 +0000 8, 23 -- User-Agent: Microsoft-Entourage/11.2.5.060620 8, 23 -- Date: Mon, 18 Sep 2006 10:38:34 -0400 8, 23 -- Subject: Re: [Microscopy] BSE image response question 8, 23 -- From: "Hull, David R" {David.R.Hull-at-nasa.gov} 8, 23 -- To: {NWWhite-at-bwxt.com} , {Microscopy-at-microscopy.com} 8, 23 -- Message-ID: {C13427AA.9B8%David.R.Hull-at-nasa.gov} 8, 23 -- Thread-Topic: [Microscopy] BSE image response question 8, 23 -- Thread-Index: AcbbMB5OXNw9YEcjEduzOwAKldSx3A== 8, 23 -- In-Reply-To: {200609181349.k8IDnl0f032128-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: 18 Sep 2006 14:17:54.0654 (UTC) FILETIME=[3B991FE0:01C6DB2D] ==============================End of - Headers==============================
Orientation is the same. No raster rotate used. In general, I tired to use identical operating parameters.
Woody
-----Original Message----- X-from: Carl Henderson [mailto:chender-at-umich.edu] Sent: Monday, September 18, 2006 10:04 AM To: White, Woody N.
Mostly we spent our time figuring out how to appease administrators who make us spend gross amounts of time justifying our budget at the expense of actually doing microscopy.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. URL: http://www.aecom.yu.edu/aif/
==============================Original Headers============================== 3, 29 -- From cammer-at-aecom.yu.edu Mon Sep 18 10:14:33 2006 3, 29 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 3, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8IFEX0g030720 3, 29 -- for {microscopy-at-microscopy.com} ; Mon, 18 Sep 2006 10:14:33 -0500 3, 29 -- Received: from draco.aecom.yu.edu (draco.aecom.yu.edu [129.98.1.160]) 3, 29 -- by mailgw.aecom.yu.edu (8.12.11.20060308/8.12.11) with ESMTP id k8IFEXED008749 3, 29 -- for {microscopy-at-microscopy.com} ; Mon, 18 Sep 2006 11:14:33 -0400 3, 29 -- Received: from draco.aecom.yu.edu (unknown [127.0.0.1]) 3, 29 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id B2213329A00 3, 29 -- for {microscopy-at-microscopy.com} ; Mon, 18 Sep 2006 11:12:16 -0400 (EDT) 3, 29 -- X-AuditID: 816201a0-a9252bb00000551c-df-450eb750a62b 3, 29 -- Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.100]) 3, 29 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id 812D62E5724 3, 29 -- for {microscopy-at-microscopy.com} ; Mon, 18 Sep 2006 11:12:16 -0400 (EDT) 3, 29 -- Received: from AIF3.aecom.yu.edu (aif3.aif.aecom.yu.edu [129.98.80.70]) 3, 29 -- by post.aecom.yu.edu (Postfix) with ESMTP id A173E25 3, 29 -- for {microscopy-at-microscopy.com} ; Mon, 18 Sep 2006 11:14:32 -0400 (EDT) 3, 29 -- Message-Id: {5.2.1.1.2.20060918111319.03083220-at-mailserver.aecom.yu.edu} 3, 29 -- X-Sender: cammer-at-mailserver.aecom.yu.edu 3, 29 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 3, 29 -- Date: Mon, 18 Sep 2006 11:14:32 -0400 3, 29 -- To: microscopy-at-microscopy.com 3, 29 -- From: Michael Cammer {cammer-at-aecom.yu.edu} 3, 29 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: What does a 3, 29 -- microscopist do all day? 3, 29 -- In-Reply-To: {200609181323.k8IDNKea022495-at-ns.microscopy.com} 3, 29 -- Mime-Version: 1.0 3, 29 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 3, 29 -- X-Brightmail-Tracker: AAAAAA== ==============================End of - Headers==============================
The opinion of the writer does not necessarily reflect the opinion of Intel Corporation.
-----Original Message----- X-from: twigg-at-estd.nrl.navy.mil [mailto:twigg-at-estd.nrl.navy.mil] Sent: Monday, September 18, 2006 5:27 AM To: Mardinly, John
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 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] Ghosted Image Window in Digital Micrograph
Question: The Dell Dimension 8400 (running Windows XP) on which I am running Digital Micrograph recently crashed. After the crash the Image widow in Digital Micrograph has been ghosted, so that I cannot insert my Ultrascan CCD camera or attempt to record an image. I have rebooted the PC a number of times, but to no avail. Perhaps the PC-savy people among you have some suggestions for me here. I have mostly used Macs and am therefore not familiar with PC ailments. Also, would it help to restart or reset the First Light Digital Camera controller?
To wash ultrathin windows (much more fragile than Be) I use Vertel XF from Du Pont. It will not dissolve adhesive. Using pipette put several drops of Vertel on the metal rim of window, so it will flow across window's surface; do not submerge or spray it, windows like gentle handling.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 371 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
It is a great question and it is tempting to be funny in the answer, but I will resist.
I agree that it would be difficult to give a general picture since microscopists come from so many different fields of study.
However I would like to say this about myself - What I do every day (amongst other things) is marvel at how incredible nature is when magnified to reveal otherwise hidden detail. I have been involved in electron microscopy since 1958 and I still feel a thrill of excitement every time I pop a specimen in the scope and turn on the beam.
Makes the many frustrations worthwhile.
Ted Dunn The EMscope Company Thailand
--- sheila.blouin-at-umb.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 } ---------------------------------------------------------------------------- } } Below is the result of your feedback form } (NJZFM-ultra-55). It was submitted by } (sheila.blouin-at-umb.edu) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } on Sunday, September 17, 2006 at 21:25:08 } --------------------------------------------------------------------------- } } Email: sheila.blouin-at-umb.edu } Name: Sheila } } Organization: University of Massachusetts, Boston } } Education: Undergraduate College } } Location: Boston, MA, USA } } Question: What does a microscopist do all day? What } kind of education do you have to have? } } --------------------------------------------------------------------------- } } ==============================Original } Headers============================== } 7, 12 -- From zaluzec-at-ultra5.microscopy.com Mon Sep } 18 07:27:51 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 } k8ICRo1f010629 } 7, 12 -- for {microscopy-at-microscopy.com} ; Mon, 18 } Sep 2006 07:27:50 -0500 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } (Unverified) } 7, 12 -- Message-Id: } {p06110401c134412a522c-at-[206.69.208.22]} } 7, 12 -- Date: Mon, 18 Sep 2006 07:27:49 -0500 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: sheila.blouin-at-umb.edu (by way of } Ask-A-Microscopist) } 7, 12 -- Subject: AskAMicroscopist: What does a } microscopist do all day? } 7, 12 -- Content-Type: text/plain; } charset="us-ascii" } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 9, 20 -- From drteddunne-at-yahoo.com Mon Sep 18 13:38:02 2006 9, 20 -- Received: from web33409.mail.mud.yahoo.com (web33409.mail.mud.yahoo.com [68.142.206.141]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k8IIc1ld001193 9, 20 -- for {microscopy-at-microscopy.com} ; Mon, 18 Sep 2006 13:38:01 -0500 9, 20 -- Received: (qmail 38107 invoked by uid 60001); 18 Sep 2006 18:38:01 -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=CfsMMRWxqS6NEPjXQcB6mycN+ijivvtaYM2cJSBGAcUujHFzVkYZO6wu+brhzjB2exDiGEcICzZFT86QzH9PFrSS/VkkMVmLjggbCEztwz1+O5kAtL/QUYNJLhNhKrYfq3oClrSk78/pNEuaubu56p6Jbj9krVGPQAZN0RA88O0= ; 9, 20 -- Message-ID: {20060918183801.38105.qmail-at-web33409.mail.mud.yahoo.com} 9, 20 -- Received: from [203.190.250.104] by web33409.mail.mud.yahoo.com via HTTP; Mon, 18 Sep 2006 11:38:01 PDT 9, 20 -- Date: Mon, 18 Sep 2006 11:38:01 -0700 (PDT) 9, 20 -- From: ted dunn {drteddunne-at-yahoo.com} 9, 20 -- Subject: AskAMicroscopist: What does a microscopist do all day? 9, 20 -- To: sheila.blouin-at-umb.edu 9, 20 -- Cc: microscopy-at-microscopy.com 9, 20 -- In-Reply-To: {200609181234.k8ICY1Go026479-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 would echo the other comment and wonder if an oxide has formed or if you have otherwise differentially affected the sample. Do you have an x-ray map for the area? Are the areas of the same composition so that contrast is only due to orientation? Are the samples taken with the same amplifier settings (i.e., contrast and brightness)?
Warren
-----Original Message----- X-from: NWWhite-at-bwxt.com [mailto:NWWhite-at-bwxt.com] Sent: Monday, September 18, 2006 8:50 AM To: wesaia-at-iastate.edu
Hello All,
I am trying to understand what is happening with a set of BSE images. Your comments will be welcome!
Below are links to two images. The first (1.5 Mb) shows two BSE images of a nickel based super alloy (Ni-Cr-Fe-Ti). Both were acquired using a 4-diode detector, 5 kV. beam, and as close to zero degrees tilt as I could set the stage. The top of the first image is in the "as polished" condition, the lower portion of the image is after a very light electro-etch. Notice the difference in channeling contrast. Z-contrast seems largely unaffected (e.g. Ti and Cr carbide inclusions). Perhaps the difference is from my inability to set EXACTLY the same tilt, but they should be within a few degrees (or better) of the same value.
Why the dramatic reversal of contrast for some grains????
The second image is simply a 60 degree tilt SE image of the same general area to show relief of the carbides due to both polishing and the etch. ...Not much.
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 expectedend-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: expectedend-at-yahoo.com Name: Jireh
Organization: SBMC
Title-Subject: [Filtered] EM from a paraffin slide
Question: We are doing clinical EM. Does anybody have a protocol for specimen prepraration for TEM from a paraffin slide?
What a great puzzler. Have you tried tilting on purpose? Perhaps going through a tilt series would be informative. One degree increments, or even half a degree could show significant changes in grey level of some grains. Just a thought.
--John
John Chandler jpchandl-at-mines.edu
-------------- Original message ---------------------- X-from: NWWhite-at-bwxt.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 } ---------------------------------------------------------------------------- } } Hello All, } } I am trying to understand what is happening with a set of BSE images. } Your comments will be welcome! } } Below are links to two images. The first (1.5 Mb) shows two BSE images } of a nickel based super alloy (Ni-Cr-Fe-Ti). Both were acquired using a } 4-diode detector, 5 kV. beam, and as close to zero degrees tilt as I } could set the stage. The top of the first image is in the "as polished" } condition, the lower portion of the image is after a very light } electro-etch. Notice the difference in channeling contrast. Z-contrast } seems largely unaffected (e.g. Ti and Cr carbide inclusions). Perhaps } the difference is from my inability to set EXACTLY the same tilt, but } they should be within a few degrees (or better) of the same value. } } Why the dramatic reversal of contrast for some grains???? } } The second image is simply a 60 degree tilt SE image of the same general } area to show relief of the carbides due to both polishing and the etch. } ...Not much. } } } http://www.bwxt.com/operations/images/sem/126867_859.jpg } } } http://www.bwxt.com/operations/images/sem/126866.jpg } } } Thanks, } Woody White } BWXT Services } Lynchburg, VA
==============================Original Headers============================== 7, 17 -- From johnpchandler-at-comcast.net Mon Sep 18 18:33:48 2006 7, 17 -- Received: from rwcrmhc15.comcast.net (rwcrmhc15.comcast.net [204.127.192.85]) 7, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8INXmt8005680 7, 17 -- for {Microscopy-at-msa.microscopy.com} ; Mon, 18 Sep 2006 18:33:48 -0500 7, 17 -- Received: from rmailcenter98.comcast.net ([204.127.197.198]) 7, 17 -- by comcast.net (rwcrmhc15) with SMTP 7, 17 -- id {20060918233347m1500nq2u2e} ; Mon, 18 Sep 2006 23:33:47 +0000 7, 17 -- Received: from [138.67.28.173] by rmailcenter98.comcast.net; 7, 17 -- Mon, 18 Sep 2006 23:33:47 +0000 7, 17 -- From: johnpchandler-at-comcast.net 7, 17 -- To: NWWhite-at-bwxt.com 7, 17 -- Cc: Microscopy-at-msa.microscopy.com 7, 17 -- Subject: Re: [Microscopy] BSE image response question 7, 17 -- Date: Mon, 18 Sep 2006 23:33:47 +0000 7, 17 -- Message-Id: {091820062333.12390.450F2CDB0000D1C80000306622135753339D0A040B020E080C9F02080106-at-comcast.net} 7, 17 -- X-Mailer: AT&T Message Center Version 1 (Apr 11 2006) 7, 17 -- X-Authenticated-Sender: am9obnBjaGFuZGxlckBjb21jYXN0Lm5ldA== ==============================End of - Headers==============================
X-from: germaine_g_boucher-at-groton.pfizer.com (by way of MicroscopyListserver)
Woody
It looks as the cristallographic contrast would dominate on chemical contrast. As John proposed, try with tilting. Channeling is very sensitif to smale angle tilting, half a degree to a few degree. If the contrast changes with so smale angles, it's channelling. Than try with higer energy.
An other question : I've never worked with a 4 sector BSE detector, but people from FEI talked me from artifacts arasing on these. Can you work in two sector mode, combining the four sectors in two pairs ? Try with different pairs. Maybe it helps to understand what happens.
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
johnpchandler-at-comcast.net a écrit : } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } Woody, } } What a great puzzler. Have you tried tilting on purpose? Perhaps going through a tilt series would be informative. One degree increments, or even half a degree could show significant changes in grey level of some grains. Just a thought. } } --John } } John Chandler } jpchandl-at-mines.edu } } } } -------------- Original message ---------------------- } X-from: NWWhite-at-bwxt.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 } } ---------------------------------------------------------------------------- } } } } Hello All, } } } } I am trying to understand what is happening with a set of BSE images. } } Your comments will be welcome! } } } } Below are links to two images. The first (1.5 Mb) shows two BSE images } } of a nickel based super alloy (Ni-Cr-Fe-Ti). Both were acquired using a } } 4-diode detector, 5 kV. beam, and as close to zero degrees tilt as I } } could set the stage. The top of the first image is in the "as polished" } } condition, the lower portion of the image is after a very light } } electro-etch. Notice the difference in channeling contrast. Z-contrast } } seems largely unaffected (e.g. Ti and Cr carbide inclusions). Perhaps } } the difference is from my inability to set EXACTLY the same tilt, but } } they should be within a few degrees (or better) of the same value. } } } } Why the dramatic reversal of contrast for some grains???? } } } } The second image is simply a 60 degree tilt SE image of the same general } } area to show relief of the carbides due to both polishing and the etch. } } ...Not much. } } } } } } http://www.bwxt.com/operations/images/sem/126867_859.jpg } } } } } } http://www.bwxt.com/operations/images/sem/126866.jpg } } } } } } Thanks, } } Woody White } } BWXT Services } } Lynchburg, VA } } } } ==============================Original Headers============================== } 7, 17 -- From johnpchandler-at-comcast.net Mon Sep 18 18:33:48 2006 } 7, 17 -- Received: from rwcrmhc15.comcast.net (rwcrmhc15.comcast.net [204.127.192.85]) } 7, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8INXmt8005680 } 7, 17 -- for {Microscopy-at-msa.microscopy.com} ; Mon, 18 Sep 2006 18:33:48 -0500 } 7, 17 -- Received: from rmailcenter98.comcast.net ([204.127.197.198]) } 7, 17 -- by comcast.net (rwcrmhc15) with SMTP } 7, 17 -- id {20060918233347m1500nq2u2e} ; Mon, 18 Sep 2006 23:33:47 +0000 } 7, 17 -- Received: from [138.67.28.173] by rmailcenter98.comcast.net; } 7, 17 -- Mon, 18 Sep 2006 23:33:47 +0000 } 7, 17 -- From: johnpchandler-at-comcast.net } 7, 17 -- To: NWWhite-at-bwxt.com } 7, 17 -- Cc: Microscopy-at-msa.microscopy.com } 7, 17 -- Subject: Re: [Microscopy] BSE image response question } 7, 17 -- Date: Mon, 18 Sep 2006 23:33:47 +0000 } 7, 17 -- Message-Id: {091820062333.12390.450F2CDB0000D1C80000306622135753339D0A040B020E080C9F02080106-at-comcast.net} } 7, 17 -- X-Mailer: AT&T Message Center Version 1 (Apr 11 2006) } 7, 17 -- X-Authenticated-Sender: am9obnBjaGFuZGxlckBjb21jYXN0Lm5ldA== } ==============================End of - Headers============================== }
==============================Original Headers============================== 10, 29 -- From jacques.faerber-at-ipcms.u-strasbg.fr Tue Sep 19 03:03:30 2006 10, 29 -- Received: from mailhost.u-strasbg.fr (mailhost.u-strasbg.fr [130.79.200.155]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8J83TpR003570 10, 29 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 03:03:29 -0500 10, 29 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr [130.79.210.2]) 10, 29 -- by mailhost.u-strasbg.fr (8.13.6/jtpda-5.5pre1) with ESMTP id k8J83SHe035808 10, 29 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 10:03:28 +0200 (CEST) 10, 29 -- Received: from [130.79.54.3] (odhinn.u-strasbg.fr [130.79.54.3]) 10, 29 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id 4E3351000121 10, 29 -- for {Microscopy-at-Microscopy.Com} ; Tue, 19 Sep 2006 10:03:08 +0200 (CEST) 10, 29 -- Message-ID: {450FA442.8090606-at-ipcms.u-strasbg.fr} 10, 29 -- Date: Tue, 19 Sep 2006 10:03:14 +0200 10, 29 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} 10, 29 -- User-Agent: Thunderbird 1.5.0.5 (X11/20060728) 10, 29 -- MIME-Version: 1.0 10, 29 -- To: Microscopy-at-microscopy.com 10, 29 -- Subject: Re: BSE image response question 10, 29 -- References: {200609182339.k8INd2dM015810-at-ns.microscopy.com} 10, 29 -- In-Reply-To: {200609182339.k8INd2dM015810-at-ns.microscopy.com} 10, 29 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-IPCMS-MailScanner: Found to be clean 10, 29 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr 10, 29 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-2.0.2 (mailhost.u-strasbg.fr [130.79.200.155]); Tue, 19 Sep 2006 10:03:28 +0200 (CEST) 10, 29 -- X-Virus-Scanned: ClamAV 0.88.4/1895/Tue Sep 19 04:49:45 2006 on mr5.u-strasbg.fr 10, 29 -- X-Virus-Status: Clean 10, 29 -- X-Spam-Status: No, score=0.2 required=5.0 tests=AWL,URI_NOVOWEL 10, 29 -- autolearn=disabled version=3.1.4 10, 29 -- X-Spam-Checker-Version: SpamAssassin 3.1.4 (2006-07-25) on mr5.u-strasbg.fr ==============================End of - Headers==============================
Can you repeat these 2 images? If so, I'd suggest duplicating this, while being particularly careful of the conditions. That is, I have seen a BSED flip its BEI contrast for different beam currents. Which is still a question in my mind why it happened, but it did happen with a Cameca multichannel (5-pair) BSED, and I watched the BEI response flip in going from 15 to ~20nA. I thought at the time it must have been a fluke with the BEI video amplifier.
On another note, can you can play with the effect of tilt by rotating the stage?
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
} I am trying to understand what is happening with a set of BSE images. } Your comments will be welcome! } } Below are links to two images. The first (1.5 Mb) shows two } BSE images of a nickel based super alloy (Ni-Cr-Fe-Ti). Both } were acquired using a 4-diode detector, 5 kV. beam, and as } close to zero degrees tilt as I could set the stage. The top } of the first image is in the "as polished" } condition, the lower portion of the image is after a very } light electro-etch. Notice the difference in channeling } contrast. Z-contrast seems largely unaffected (e.g. Ti and } Cr carbide inclusions). Perhaps the difference is from my } inability to set EXACTLY the same tilt, but they should be } within a few degrees (or better) of the same value. } } Why the dramatic reversal of contrast for some grains???? } } The second image is simply a 60 degree tilt SE image of the } same general area to show relief of the carbides due to both } polishing and the etch. } ...Not much. } } } http://www.bwxt.com/operations/images/sem/126867_859.jpg } } } http://www.bwxt.com/operations/images/sem/126866.jpg } } } Thanks, } Woody White } BWXT Services } Lynchburg, VA } } } } } ==============================Original } Headers============================== } 14, 26 -- From nwwhite-at-bwxt.com Mon Sep 18 08:48:46 2006 } 14, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) } 14, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) } with ESMTP id k8IDmkt4030869 } 14, 26 -- for {microscopy-at-msa.microscopy.com} ; Mon, 18 } Sep 2006 08:48:46 -0500 } 14, 26 -- Received: from ([131.184.13.224]) } 14, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.2215552; } 14, 26 -- Mon, 18 Sep 2006 09:48:37 -0400 } 14, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET } ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with } Microsoft SMTPSVC(6.0.3790.1830); } 14, 26 -- Mon, 18 Sep 2006 09:48:37 -0400 } 14, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 14, 26 -- Content-class: urn:content-classes:message } 14, 26 -- MIME-Version: 1.0 } 14, 26 -- Content-Type: text/plain; } 14, 26 -- charset="US-ASCII" } 14, 26 -- Subject: BSE image response question } 14, 26 -- Date: Mon, 18 Sep 2006 09:48:37 -0400 } 14, 26 -- Message-ID: } {360DDAABED436B49BA397357CCEE8BB703D063-at-BWXSPO01.BWXS.BWXTECH.NET} } 14, 26 -- X-MS-Has-Attach: } 14, 26 -- X-MS-TNEF-Correlator: } 14, 26 -- Thread-Topic: BSE image response question } 14, 26 -- Thread-Index: AcbbKSRJZ3PRklUEQNiswlKEqWhmhg== } 14, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} } 14, 26 -- To: "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} } 14, 26 -- X-OriginalArrivalTime: 18 Sep 2006 13:48:37.0909 } (UTC) FILETIME=[247F1C50:01C6DB29] } 14, 26 -- Content-Transfer-Encoding: 8bit } 14, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit } by ns.microscopy.com id k8IDmkt4030869 } ==============================End of - } Headers============================== }
Has anyone got any opinions on the Fullam, Deben (Gatan) and Kammrath and Weiss strainign stages for the SEM, which is the best? Which is the best bang for the buck? Are there any other manufacturers? Any help welcome. Thanks.
-- John Mansfield PhD Cphys CSci MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 USA Phone: (734) 936-3352 FAX (734) 763-2282 Cell. Phone: (734) 834-3913 (Leaving a phone message at 936-3352 is preferable to 834-3913) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42° 16' 48" Long. 83° 43' 48" AIM: thejfmjfm Yahoo: thejfmjfm Skype: thejfmjfm
Home address: 4304 Spring Lake Boulevard Ann Arbor MI 48108-9657 Phone (734) 994-3096
Please note: Electronic Mail is not secure, but should be read several times every day, and should definiely be used for urgent or sensitive issues.
==============================Original Headers============================== 8, 22 -- From jfmjfm-at-umich.edu Tue Sep 19 09:41:53 2006 8, 22 -- Received: from srvr20.engin.umich.edu (srvr20.engin.umich.edu [141.213.75.22]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8JEfrmU032664 8, 22 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 09:41:53 -0500 8, 22 -- Received: from smtp.engin.umich.edu (root-at-smtp.engin.umich.edu [141.213.75.24]) 8, 22 -- by srvr20.engin.umich.edu (8.13.6/8.13.6) with ESMTP id k8JEfngV022783 8, 22 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 10:41:53 -0400 (EDT) 8, 22 -- Received: from [192.168.2.4] (c-68-40-243-101.hsd1.mi.comcast.net [68.40.243.101]) 8, 22 -- (authenticated bits=0) 8, 22 -- by smtp.engin.umich.edu (8.13.6/8.13.6) with ESMTP id k8JEfm1d010635 8, 22 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NO) 8, 22 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 10:41:48 -0400 (EDT) 8, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 22 -- Message-Id: {573681CC-0E43-4B17-BD0A-9D387FE49F92-at-umich.edu} 8, 22 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 8, 22 -- To: microscopy-at-microscopy.com 8, 22 -- From: John Mansfield {jfmjfm-at-umich.edu} 8, 22 -- Subject: Straining stages for the SEM 8, 22 -- Date: Tue, 19 Sep 2006 10:41:56 -0400 8, 22 -- X-Mailer: Apple Mail (2.752.2) 8, 22 -- Content-Transfer-Encoding: 8bit 8, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8JEfrmU032664 ==============================End of - Headers==============================
A feedback of any experience with the Hitachi TM1000 low vacuum desktop scanning electron microscope regarding resolution, performance, reliability, costs, etc. will be appreciated.
Thank you,
Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
tel 951 827 2998 fax 951 827 2489 bozhilov-at-ucr.edu
==============================Original Headers============================== 7, 19 -- From bozhilov-at-ucr.edu Tue Sep 19 10:01:12 2006 7, 19 -- Received: from sentry.ucr.edu (sentry.ucr.edu [138.23.226.224]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8JF1Acd011010 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 10:01:11 -0500 7, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 7, 19 -- by sentry.ucr.edu (MOS 3.5.9-GR) 7, 19 -- with ESMTP id DZU42628 (AUTH via LOGINBEFORESMTP) 7, 19 -- for {microscopy-at-microscopy.com} ; 7, 19 -- Tue, 19 Sep 2006 08:01:04 -0700 (PDT) 7, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 7, 19 -- Content-Transfer-Encoding: 7bit 7, 19 -- Message-Id: {85F81D99-9C1B-4C52-A0F0-96C3A80051D4-at-ucr.edu} 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 7, 19 -- Subject: Hitachi TM1000 7, 19 -- Date: Tue, 19 Sep 2006 08:01:04 -0700 7, 19 -- X-Mailer: Apple Mail (2.752.2) 7, 19 -- X-Junkmail-Status: score=0/65, host=sentry.ucr.edu ==============================End of - Headers==============================
Krassimir: I've done a little work on this fun little SEM which bridges the gap between light microscopy and high-resolution SEM. It's very easy to use as it has a fixed working distance, fixed apertures, and a fixed kV (15kV). It's a tungsten gun with a maximum magnification of 10kX. I'm not sure what the resolution spec actually is but it seems to be adequate for at least 0.5 um. The only facilities it needs is 110-115V in a regular 3-prong (USA) wall outlet. It needs no compressed air/nitrogen or water. It connects to any computer with a USB socket and the user interface and imaging software is very easy to use. I understand from the rep that it's a big hit with elementary kids as they can image their own samples with minimal instruction. I don't have any feel for the reliability because it hasn't been on the market that long but there's not a lot to go wrong with it, other than filament changes. The price seems to be around $US 65K. If you supply your own computer it's a bit less. Of course it has no EDX detector port or any other detector ports. I have no financial interest in this tool or Hitachi. I just think it's way cool and wish I had one on my desk.
bozhilov-at-ucr.edu wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } A feedback of any experience with the Hitachi TM1000 low vacuum } desktop scanning electron microscope regarding resolution, } performance, reliability, costs, etc. will be appreciated. } } Thank you, } } Krassimir N. Bozhilov } Central Facility for Advanced Microscopy and Microanalysis } University of California } Riverside, CA 92521 } } tel 951 827 2998 } fax 951 827 2489 } bozhilov-at-ucr.edu } } } } } ==============================Original Headers============================== } 7, 19 -- From bozhilov-at-ucr.edu Tue Sep 19 10:01:12 2006 } 7, 19 -- Received: from sentry.ucr.edu (sentry.ucr.edu [138.23.226.224]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8JF1Acd011010 } 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 10:01:11 -0500 } 7, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) } 7, 19 -- by sentry.ucr.edu (MOS 3.5.9-GR) } 7, 19 -- with ESMTP id DZU42628 (AUTH via LOGINBEFORESMTP) } 7, 19 -- for {microscopy-at-microscopy.com} ; } 7, 19 -- Tue, 19 Sep 2006 08:01:04 -0700 (PDT) } 7, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 7, 19 -- Content-Transfer-Encoding: 7bit } 7, 19 -- Message-Id: {85F81D99-9C1B-4C52-A0F0-96C3A80051D4-at-ucr.edu} } 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 7, 19 -- To: microscopy-at-microscopy.com } 7, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} } 7, 19 -- Subject: Hitachi TM1000 } 7, 19 -- Date: Tue, 19 Sep 2006 08:01:04 -0700 } 7, 19 -- X-Mailer: Apple Mail (2.752.2) } 7, 19 -- X-Junkmail-Status: score=0/65, host=sentry.ucr.edu } ==============================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, 23 -- From r-holdford-at-ti.com Tue Sep 19 10:56:02 2006 5, 23 -- Received: from bear.ext.ti.com (bear.ext.ti.com [192.94.94.41]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8JFu2dk022392 5, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 19 Sep 2006 10:56:02 -0500 5, 23 -- Received: from dlep32.itg.ti.com ([157.170.170.70]) 5, 23 -- by bear.ext.ti.com (8.13.7/8.13.7) with ESMTP id k8JFttrx006886 5, 23 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO); 5, 23 -- Tue, 19 Sep 2006 10:56:00 -0500 5, 23 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 5, 23 -- by dlep32.itg.ti.com (8.13.7/8.13.7) with ESMTP id k8JFtsum029506; 5, 23 -- Tue, 19 Sep 2006 10:55:54 -0500 (CDT) 5, 23 -- Message-ID: {4510130B.6070805-at-ti.com} 5, 23 -- Date: Tue, 19 Sep 2006 10:55:55 -0500 5, 23 -- From: Becky Holdford {r-holdford-at-ti.com} 5, 23 -- Organization: SC Packaging Development -- FA Development 5, 23 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 5, 23 -- MIME-Version: 1.0 5, 23 -- To: bozhilov-at-ucr.edu, MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 23 -- Subject: Re: [Microscopy] Hitachi TM1000 5, 23 -- References: {200609191501.k8JF1Qbq011327-at-ns.microscopy.com} 5, 23 -- In-Reply-To: {200609191501.k8JF1Qbq011327-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==============================
We just had one installed two weeks ago in our lab. We haven't really played with it much. But it is indeed a nice tool, doesn't take up much space, pumps down very quickly and fairly easy to use. You can insert fresh biological samples and image it. The max magnification you can go to is 10k. We placed it on a heavy microtome table to minimize any vibration problem. You can contact Marine Reef International (Paul DeGeorge) in LA area and get some more info. I believe they are the Hitachi distributor for this instrument.
No financial interest in Hitachi or Marine Reef.
Soumitra
Quoting bozhilov-at-ucr.edu:
} } } } ---------------------------------------------------------------------------- } 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 feedback of any experience with the Hitachi TM1000 low vacuum } desktop scanning electron microscope regarding resolution, } performance, reliability, costs, etc. will be appreciated. } } Thank you, } } Krassimir N. Bozhilov } Central Facility for Advanced Microscopy and Microanalysis } University of California } Riverside, CA 92521 } } tel 951 827 2998 } fax 951 827 2489 } bozhilov-at-ucr.edu } } } } } ==============================Original } Headers============================== } 7, 19 -- From bozhilov-at-ucr.edu Tue Sep 19 10:01:12 2006 } 7, 19 -- Received: from sentry.ucr.edu (sentry.ucr.edu [138.23.226.224]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k8JF1Acd011010 } 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 10:01:11 -0500 } 7, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu } [138.23.185.162]) } 7, 19 -- by sentry.ucr.edu (MOS 3.5.9-GR) } 7, 19 -- with ESMTP id DZU42628 (AUTH via LOGINBEFORESMTP) } 7, 19 -- for {microscopy-at-microscopy.com} ; } 7, 19 -- Tue, 19 Sep 2006 08:01:04 -0700 (PDT) } 7, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 7, 19 -- Content-Transfer-Encoding: 7bit } 7, 19 -- Message-Id: {85F81D99-9C1B-4C52-A0F0-96C3A80051D4-at-ucr.edu} } 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; } format=flowed } 7, 19 -- To: microscopy-at-microscopy.com } 7, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} } 7, 19 -- Subject: Hitachi TM1000 } 7, 19 -- Date: Tue, 19 Sep 2006 08:01:04 -0700 } 7, 19 -- X-Mailer: Apple Mail (2.752.2) } 7, 19 -- X-Junkmail-Status: score=0/65, host=sentry.ucr.edu } ==============================End of - } Headers============================== }
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
I was working as a research assistant in the electron microscopy lab of Edinburgh University Zoology Department in Scotland. This was Scotland's first TEM, a beautiful space-age looking Siemens Elmiskop 1b.(It was actually installed in 1956 not 58). The pumping system was all controlled with manual valves (had a mercury pump and an oil diff pump if I remember correctly) and the lens and gun alignments were all manual. It was quite a task to align after a filament change.
This TEM produced excellent images and we could routinely get about 12A resolution though it was capable of better.
We had frequent visits from German engineers to begin with but gradually tamed the instrument. The electronics of course were all glass tubes which I liked because once the circuits were understood it was comparatively easy to locate faults.
Many of the specimen techniques we had to develop ourselves since there wasn't much in the literature. I was very fortunate to have David Bradley work in our department for a while since he pioneered carbon support films which revolutionized support film technology.
For embedding we initially had no choice but methacrylate - not all that good but it was still exciting to get those pictures of cellular structure and you could get published anywhere just because the technology was so new!
It was fun and satisfying.
Ted
--- "David L. Jones" {dljones-at-bestweb.net} wrote:
} Ted, } } In 1958, electron microscopy was very difficult to } do...Where were you } working at that time? What microscope did you work } on? Now those were early } machines... I'd love to hear about working with such } an early instrument. } My first SEM experience was on an SEM that required } manual alignment of } the lenses... } } dj } } On Mon, 18 Sep 2006, 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 } } } ---------------------------------------------------------------------------- } } } } It is a great question and it is tempting to be } funny } } in the answer, but I will resist. } } } } I agree that it would be difficult to give a } general } } picture since microscopists come from so many } } different fields of study. } } } } However I would like to say this about myself - } What I } } do every day (amongst other things) is marvel at } how } } incredible nature is when magnified to reveal } } otherwise hidden detail. I have been involved in } } electron microscopy since 1958 and I still feel a } } thrill of excitement every time I pop a specimen } in } } the scope and turn on the beam. } } } } Makes the many frustrations worthwhile. } } } } Ted Dunn } } The EMscope Company } } Thailand } } } } --- sheila.blouin-at-umb.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 } } } } } } ---------------------------------------------------------------------------- } } } } } } Below is the result of your feedback form } } } (NJZFM-ultra-55). It was submitted by } } } (sheila.blouin-at-umb.edu) from } } } } } } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } } } on Sunday, September 17, 2006 at 21:25:08 } } } } } } --------------------------------------------------------------------------- } } } } } } Email: sheila.blouin-at-umb.edu } } } Name: Sheila } } } } } } Organization: University of Massachusetts, Boston } } } } } } Education: Undergraduate College } } } } } } Location: Boston, MA, USA } } } } } } Question: What does a microscopist do all day? } What } } } kind of education do you have to have? } } } } } } } } } --------------------------------------------------------------------------- } } } } } } ==============================Original } } } Headers============================== } } } 7, 12 -- From zaluzec-at-ultra5.microscopy.com Mon } Sep } } } 18 07:27:51 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 } } } k8ICRo1f010629 } } } 7, 12 -- for {microscopy-at-microscopy.com} ; Mon, } 18 } } } Sep 2006 07:27:50 -0500 } } } 7, 12 -- Mime-Version: 1.0 } } } 7, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com } } } (Unverified) } } } 7, 12 -- Message-Id: } } } {p06110401c134412a522c-at-[206.69.208.22]} } } } 7, 12 -- Date: Mon, 18 Sep 2006 07:27:49 -0500 } } } 7, 12 -- To: microscopy-at-microscopy.com } } } 7, 12 -- From: sheila.blouin-at-umb.edu (by way of } } } Ask-A-Microscopist) } } } 7, 12 -- Subject: AskAMicroscopist: What does a } } } microscopist do all day? } } } 7, 12 -- 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 } } } } ==============================Original } Headers============================== } } 9, 20 -- From drteddunne-at-yahoo.com Mon Sep 18 } 13:38:02 2006 } } 9, 20 -- Received: from } web33409.mail.mud.yahoo.com } (web33409.mail.mud.yahoo.com [68.142.206.141]) } } 9, 20 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } k8IIc1ld001193 } } 9, 20 -- for {microscopy-at-microscopy.com} ; Mon, 18 } Sep 2006 13:38:01 -0500 } } 9, 20 -- Received: (qmail 38107 invoked by uid } 60001); 18 Sep 2006 18:38:01 -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=CfsMMRWxqS6NEPjXQcB6mycN+ijivvtaYM2cJSBGAcUujHFzVkYZO6wu+brhzjB2exDiGEcICzZFT86QzH9PFrSS/VkkMVmLjggbCEztwz1+O5kAtL/QUYNJLhNhKrYfq3oClrSk78/pNEuaubu56p6Jbj9krVGPQAZN0RA88O0= } ; } } 9, 20 -- Message-ID: } {20060918183801.38105.qmail-at-web33409.mail.mud.yahoo.com} } } 9, 20 -- Received: from [203.190.250.104] by } web33409.mail.mud.yahoo.com via HTTP; Mon, 18 Sep } 2006 11:38:01 PDT } } 9, 20 -- Date: Mon, 18 Sep 2006 11:38:01 -0700 } (PDT) } } 9, 20 -- From: ted dunn {drteddunne-at-yahoo.com} } } 9, 20 -- Subject: AskAMicroscopist: What does a } microscopist do all day? } } 9, 20 -- To: sheila.blouin-at-umb.edu } } 9, 20 -- Cc: microscopy-at-microscopy.com } } 9, 20 -- In-Reply-To: } {200609181234.k8ICY1Go026479-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============================== } } } }
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==============================Original Headers============================== 12, 20 -- From drteddunne-at-yahoo.com Tue Sep 19 11:28:49 2006 12, 20 -- Received: from web33402.mail.mud.yahoo.com (web33402.mail.mud.yahoo.com [68.142.206.134]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k8JGSluE011177 12, 20 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 11:28:48 -0500 12, 20 -- Received: (qmail 24443 invoked by uid 60001); 19 Sep 2006 16:28:45 -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=nqFB/XyCsBhorx5FPC/qNw9B84695ae762eX1iERJmHI7SZKe/vNh6C4s0vkJAA9rBl50rdRAgGxEpZmynVgE3BAsJFtyiNIQfu2+xQODZk6gFfgp/URNmOqPyDDjqotcciiWWYvr7MDhD8yU5DFzxzhEXLlVntToG5oZI5yFWs= ; 12, 20 -- Message-ID: {20060919162845.24441.qmail-at-web33402.mail.mud.yahoo.com} 12, 20 -- Received: from [203.190.250.104] by web33402.mail.mud.yahoo.com via HTTP; Tue, 19 Sep 2006 09:28:45 PDT 12, 20 -- Date: Tue, 19 Sep 2006 09:28:45 -0700 (PDT) 12, 20 -- From: ted dunn {drteddunne-at-yahoo.com} 12, 20 -- Subject: AskAMicroscopist: What does a microscopist do all day? 12, 20 -- To: "David L. Jones" {dljones-at-bestweb.net} 12, 20 -- Cc: microscopy-at-microscopy.com 12, 20 -- In-Reply-To: {Pine.WNT.4.64.0609181522561.3000-at-H-F1} 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==============================
Dear Woody, I would suspect that the reason for the difference has more to do with the removal of the thin, amorphous layer left on the as-polished sample, but I must admit that the contrast reversal is dramatic. BSE can be very strange that way and I never get the same image contrast twice on the same sample. Try tilting slightly and watch it change, particularly when you are viewing channeling contrast on a homogenous, single-phase sample. 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: {NWWhite-at-bwxt.com} To: {mager-at-interchange.ubc.ca} Sent: Monday, September 18, 2006 6:55 AM
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 martimor-at-nmsu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: martimor-at-nmsu.edu Name: Marti
Organization: NMSU
Title-Subject: [Filtered] RE: Spurrs's
Question: Just a quick question to the listserv please about Spurrís embedding media for anyone who may know. What was update (if there was one) of the Spurr's embedding media for TEM? I believe there were some changes with the new Spurrís mixture due to a new safer component added in and switched with one of the previous hazardous ingredients which caused some noticeable changes in peopleís research. Are researchers still using their old stashes and waiting still for word on the new Spurrís resin to be corrected?
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 CHudson-at-slb.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: CHudson-at-slb.com Name: Candi
Organization: Schlumberger
Title-Subject: [Filtered] Hitachi TM1000
Question: I have the same question. Would also like to know any information/feedback of any experience with the Hitachi TM1000 low vacuum desktop scanning electron microscope regarding resolution, performance, reliability, costs, etc. will be appreciated.
We had one of these in the lab for a week, have never had such a stream of visitors wanting to use the SEM! For the price, it was great. I'd echo earlier comments - it's very easy to use - no need to gold coat, quite good images (we were pushing the limits of its low vacuum capacity), not bad resolution, esp. for low mag work which is where many of our taxonomists work, compact, no need for water cooling or nitrogen venting, etc. It survived a whole stream of inexperienced users. If we'd had the spare cash, we would have been very tempted. It'd be nice with a cooled stage because the low vacuum wasn't quite low enough to prevent desiccation of the most watery specimens. Good value!
cheers, Rosemary
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre ph. 61-2-6246 5475 CSIRO Plant Industry mob. 61-0402 835 973 GPO Box 1600 fax. 61-2-6246 5334 Canberra, ACT 2601 Australia
I suppose if one could collect a secondary electron image this could be even more useful. As far as a low vacuum, backscattered imaging microscope representing a "bridge" between optical and a conventional SEM, I would say a section of the bridge is missing; namely the SEI.
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
Email: CHudson-at-slb.com Name: Candi
Organization: Schlumberger
Title-Subject: [Filtered] Hitachi TM1000
Question: I have the same question. Would also like to know any information/feedback of any experience with the Hitachi TM1000 low vacuum desktop scanning electron microscope regarding resolution, performance, reliability, costs, etc. will be appreciated.
==============================Original Headers============================== 10, 27 -- From emlabservices-at-cox.net Tue Sep 19 20:14:27 2006 10, 27 -- Received: from eastrmmtao05.cox.net (eastrmmtao05.cox.net [68.230.240.34]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8K1EQJt007362 10, 27 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 20:14:26 -0500 10, 27 -- Received: from eastrmimpo02.cox.net ([68.1.16.120]) by eastrmmtao05.cox.net 10, 27 -- (InterMail vM.6.01.06.01 201-2131-130-101-20060113) with ESMTP 10, 27 -- id {20060920011415.OVSG7951.eastrmmtao05.cox.net-at-eastrmimpo02.cox.net} 10, 27 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 21:14:15 -0400 10, 27 -- Received: from EMLabServices ([24.255.213.54]) 10, 27 -- by eastrmimpo02.cox.net with bizsmtp 10, 27 -- id QRDz1V00U1AzDvc0000000 10, 27 -- Tue, 19 Sep 2006 21:14:08 -0400 10, 27 -- Message-ID: {02ce01c6dc52$1ba2f050$6500a8c0-at-EMLabServices} 10, 27 -- From: "EM Lab Services" {emlabservices-at-cox.net} 10, 27 -- To: {Microscopy-at-microscopy.com} 10, 27 -- Subject: Hitachi TM1000 10, 27 -- Date: Tue, 19 Sep 2006 19:14:18 -0600 10, 27 -- MIME-Version: 1.0 10, 27 -- Content-Type: text/plain; 10, 27 -- format=flowed; 10, 27 -- charset="iso-8859-1"; 10, 27 -- reply-type=original 10, 27 -- Content-Transfer-Encoding: 7bit 10, 27 -- X-Priority: 3 10, 27 -- X-MSMail-Priority: Normal 10, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 10, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
Had the computer processor or camera control box been moved at all just before the problem? We had a similar experience of 'greyed-out' controls after a user moved the computer processor to find a USB point (there was an extension lead in place but they didn't notice it!) and in the process stressed the cable from the camera to its PC board - a plastic clip holding the board in place had snapped and the board was loose. On securing the board in place with a screw all was OK. We had valuable help from Gatan with the problem.
Hope this helps Regards Ursula -------------
Ursula J. Potter Centre for Electron Optical Studies (CEOS) Building 3 West 2.15 The University of Bath Claverton Down Bath BA2 7AY UK Tel: 01225 385651 Email: U.J.Potter-at-bath.ac.uk
==============================Original Headers============================== 5, 22 -- From U.J.Potter-at-bath.ac.uk Wed Sep 20 03:25:56 2006 5, 22 -- Received: from kelly.bath.ac.uk (kelly.bath.ac.uk [138.38.32.20]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8K8PtUI010013 5, 22 -- for {microscopy-at-microscopy.com} ; Wed, 20 Sep 2006 03:25:55 -0500 5, 22 -- Received: from amos.bath.ac.uk ([138.38.32.36] ident=mmdf) 5, 22 -- by kelly.bath.ac.uk with smtp id 1GPxP1-0007FX-3r 5, 22 -- (return-path {U.J.Potter-at-bath.ac.uk} ); Wed, 20 Sep 2006 09:25:55 +0100 5, 22 -- Received: from eapc-03.campus.bath.ac.uk 5, 22 -- ( eapc-03.campus.bath.ac.uk [138.38.136.63] ) by bath.ac.uk 5, 22 -- id aa15989 ; 20 Sep 2006 09:25 +0100 5, 22 -- Date: Wed, 20 Sep 2006 09:25:55 +0100 5, 22 -- From: Ursula Potter {U.J.Potter-at-bath.ac.uk} 5, 22 -- To: twigg-at-estd.nrl.navy.mil, microscopy-at-microscopy.com 5, 22 -- Subject: Ghosted Image Window in Digital Micrograph 5, 22 -- Message-ID: {1317015.1158744355-at-eapc-03.campus.bath.ac.uk} 5, 22 -- Originator-Info: login-id=mssujp; server=imaphost.bath.ac.uk 5, 22 -- X-Mailer: Mulberry/3.1.0 (Win32) 5, 22 -- MIME-Version: 1.0 5, 22 -- Content-Type: text/plain; charset=us-ascii; format=flowed 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- Content-Disposition: inline 5, 22 -- X-Scanner: 1e6ba4d679d16f1753d9cf2e37791b53f54e02e5 ==============================End of - Headers==============================
I forgot to mention when I sent the article citation: we are using up our current stock of ERL 4206 (VCD), and will change to 4221 when the current stock is gone. 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 martimor-at-nmsu.edu as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: martimor-at-nmsu.edu } Name: Marti } } Organization: NMSU } } Title-Subject: [Filtered] RE: Spurrs's } } Question: Just a quick question to the listserv } please about SpurrÌs embedding media for anyone } who may know. What was update (if there was one) } of the Spurr's embedding media for TEM? I } believe there were some changes with the new } SpurrÌs mixture due to a new safer component } added in and switched with one of the previous } hazardous ingredients which caused some } noticeable changes in peopleÌs research. Are } researchers still using their old stashes and } waiting still for word on the new SpurrÌs resin } to be corrected? } } Thanks in advance! } MM -- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 3, 25 -- From oshel1pe-at-cmich.edu Wed Sep 20 07:14:06 2006 3, 25 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8KCE5HZ025173 3, 25 -- for {Microscopy-at-microscopy.com} ; Wed, 20 Sep 2006 07:14:05 -0500 3, 25 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 25 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k8KCk3lk019006; 3, 25 -- Wed, 20 Sep 2006 08:46:15 -0400 3, 25 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 25 -- Wed, 20 Sep 2006 08:13:53 -0400 3, 25 -- Mime-Version: 1.0 3, 25 -- Message-Id: {f06230904c136e0450e3f-at-[141.209.160.249]} 3, 25 -- In-Reply-To: {200609192342.k8JNgVBu022078-at-ns.microscopy.com} 3, 25 -- References: {200609192342.k8JNgVBu022078-at-ns.microscopy.com} 3, 25 -- Date: Wed, 20 Sep 2006 08:13:53 -0400 3, 25 -- To: martimor-at-nmsu.edu 3, 25 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 25 -- Subject: Re: [Microscopy] viaWWW: Spurrs's 3, 25 -- Cc: Microscopy-at-microscopy.com 3, 25 -- Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" 3, 25 -- X-OriginalArrivalTime: 20 Sep 2006 12:13:53.0956 (UTC) FILETIME=[3D6C3640:01C6DCAE] 3, 25 -- X-CanItPRO-Stream: default 3, 25 -- X-Spam-Score: -4 () L_EXCH_MF 3, 25 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 3, 25 -- Content-Transfer-Encoding: 8bit 3, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8KCE5HZ025173 ==============================End of - Headers==============================
See the July, 2006 edition of Microscopy Today http://www.microscopy-today.com/cgi-bin/MTWWWListingSQL.pl
E. Ann Ellis, "Solutions to the Problem of=20 Substitution of ERL 4221 for Vinyl Cyclohexene=20 Dioxide in Spurr Low Viscosity Embedding=20 =46ormulations"
Phil
} --|This Question/Comment was submitted to the Microscopy Listserver } --|using the WWW based Form at=20 } --|http://microscopy.com/MicroscopyListserver/MLFormMail.html } --|--------------------------------------------------------------------------- } --|Remember this posting is most likely not from a Subscriber, so } when replyin= } g } --|please copy both martimor-at-nmsu.edu as well as the MIcroscopy Listserver } --|--------------------------------------------------------------------------- } --| } --|Email: martimor-at-nmsu.edu } --|Name: Marti } --| } --|Organization: NMSU } --| } --|Title-Subject: [Filtered] RE: Spurrs's } --| } --|Question: Just a quick question to the listserv=20 } --|please about Spurr=CCs embedding media for anyone=20 } --|who may know. What was update (if there was one)=20 } --|of the Spurr's embedding media for TEM? I=20 } --|believe there were some changes with the new=20 } --|Spurr=CCs mixture due to a new safer component=20 } --|added in and switched with one of the previous=20 } --|hazardous ingredients which caused some=20 } --|noticeable changes in people=CCs research. Are=20 } --|researchers still using their old stashes and=20 } --|waiting still for word on the new Spurr=CCs resin=20 } --|to be corrected? } --| } --|Thanks in advance! } --|MM -- Philip Oshel Technical Editor, Microscopy Today Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 fax: (989) 774-3462
==============================Original Headers============================== 5, 20 -- From oshel1pe-at-cmich.edu Wed Sep 20 07:35:47 2006 5, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8KCZks6003377 5, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 20 Sep 2006 07:35:46 -0500 5, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 5, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k8KD7qlg022158 5, 20 -- for {Microscopy-at-microscopy.com} ; Wed, 20 Sep 2006 09:07:56 -0400 5, 20 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 5, 20 -- Wed, 20 Sep 2006 08:35:41 -0400 5, 20 -- Mime-Version: 1.0 5, 20 -- Message-Id: {f0623090ac136e5d75c34-at-[141.209.160.249]} 5, 20 -- Date: Wed, 20 Sep 2006 08:35:39 -0400 5, 20 -- To: Microscopy-at-microscopy.com 5, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 5, 20 -- Subject: RE: Spurrs's 5, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 20 -- X-OriginalArrivalTime: 20 Sep 2006 12:35:43.0844 (UTC) FILETIME=[4A2D4240:01C6DCB1] 5, 20 -- X-CanItPRO-Stream: default 5, 20 -- X-Spam-Score: -2.8 () J_CHICKENPOX_53,J_CHICKENPOX_63,L_EXCH_MF 5, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
I have not read Ann's article, but after having trouble with the plastic being very brittle after switching to 4221 with the standard "firm" formulation. I experimented and found the following formulation to provide firm blocks, very reliably, that are not brittle:
ERL 4221 - 20g DER 736 - 16g NSA - 50g DMAE - .6g
The blocks are cured overnight at 70C.
Hope this helps, Steve
Steven Lee Chief Technologist Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 75246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
-----Original Message----- X-from: martimor-at-nmsu.edu [mailto:martimor-at-nmsu.edu] Sent: Tuesday, September 19, 2006 6:44 PM To: Lee, Steven
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 martimor-at-nmsu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: martimor-at-nmsu.edu Name: Marti
Organization: NMSU
Title-Subject: [Filtered] RE: Spurrs's
Question: Just a quick question to the listserv please about Spurrís embedding media for anyone who may know. What was update (if there was one) of the Spurr's embedding media for TEM? I believe there were some changes with the new Spurrís mixture due to a new safer component added in and switched with one of the previous hazardous ingredients which caused some noticeable changes in peopleís research. Are researchers still using their old stashes and waiting still for word on the new Spurrís resin to be corrected?
==============================Original Headers============================== 9, 14 -- From zaluzec-at-microscopy.com Tue Sep 19 18:31:54 2006 9, 14 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 9, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8JNVrqV007330 9, 14 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 18:31:54 -0500 9, 14 -- Mime-Version: 1.0 9, 14 -- X-Sender: (Unverified) 9, 14 -- Message-Id: {p06110400c1362e5bec01-at-[206.69.208.22]} 9, 14 -- Date: Tue, 19 Sep 2006 18:31:52 -0500 9, 14 -- To: microscopy-at-microscopy.com 9, 14 -- From: martimor-at-nmsu.edu (by way of MicroscopyListserver) 9, 14 -- Subject: viaWWW: Spurrs's 9, 14 -- Content-Type: text/plain; charset="iso-8859-1" 9, 14 -- Content-Transfer-Encoding: 8bit 9, 14 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8JNVrqV007330 ==============================End of - Headers==============================
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==============================Original Headers============================== 22, 26 -- From StevenLe-at-BaylorHealth.edu Wed Sep 20 08:19:57 2006 22, 26 -- Received: from BHDAEXIMS02.bhcs.pvt (mailhost2.baylorhealth.edu [4.22.141.163]) 22, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8KDJukR014483 22, 26 -- for {Microscopy-at-microscopy.com} ; Wed, 20 Sep 2006 08:19:56 -0500 22, 26 -- Received: from BHDAEXFE01.bhcs.pvt ([10.5.3.72]) by BHDAEXIMS02.bhcs.pvt with InterScan Messaging Security Suite; Wed, 20 Sep 2006 08:19:51 -0500 22, 26 -- Received: from BHDAEXCH11.bhcs.pvt ([10.5.3.67]) by BHDAEXFE01.bhcs.pvt with Microsoft SMTPSVC(6.0.3790.211); 22, 26 -- Wed, 20 Sep 2006 08:19:51 -0500 22, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 22, 26 -- Content-class: urn:content-classes:message 22, 26 -- MIME-Version: 1.0 22, 26 -- Content-Type: text/plain; 22, 26 -- charset="iso-8859-1" 22, 26 -- Subject: RE: [Microscopy] viaWWW: Spurrs's 22, 26 -- Date: Wed, 20 Sep 2006 08:19:50 -0500 22, 26 -- Message-ID: {74A8B7A904152141BD5AA2761F5D223401C96026-at-BHDAEXCH11.bhcs.pvt} 22, 26 -- In-Reply-To: {200609192344.k8JNiKo9027169-at-ns.microscopy.com} 22, 26 -- X-MS-Has-Attach: 22, 26 -- X-MS-TNEF-Correlator: 22, 26 -- Thread-Topic: [Microscopy] viaWWW: Spurrs's 22, 26 -- Thread-Index: AcbcRZQ902NuxxGoTXy1++xjykzFmAAcHlxQ 22, 26 -- From: "Lee, Steven" {StevenLe-at-BaylorHealth.edu} 22, 26 -- To: {martimor-at-nmsu.edu} 22, 26 -- Cc: {Microscopy-at-microscopy.com} 22, 26 -- X-OriginalArrivalTime: 20 Sep 2006 13:19:51.0339 (UTC) FILETIME=[7434EFB0:01C6DCB7] 22, 26 -- Content-Transfer-Encoding: 8bit 22, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8KDJukR014483 ==============================End of - Headers==============================
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Email: joseph_woolman-at-raytheon.com Name: Joe
Organization: Raytheon
Title-Subject: [Filtered] Used FE-SEM's
Question: Does anyone know where to purchase a "good" used FE-SEM or FE-ESEM?
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Email: miza069-at-yahoo.com Name: Christoph Mitterbauer
Organization: UC Davis/CHMS
Title-Subject: [Filtered] Assistant Professor in (S)TEM, UC-Davis
Question: The Department of Chemical Engineering and Materials Science invites applications for a tenure-track position at the Assistant Professor level in the area of advanced (scanning) transmission electron microscopy ((S)TEM). In the last two years, UC Davis has made a significant investment in electron microscopy facilities for both the engineering and biological sciences (including three new field-emission (S)TEMs). The current position seeks a candidate to work within this environment who has expertise in the development and application of advanced methods of imaging and analysis in (S)TEM for the engineering sciences, and who has a strong commitment to applying these methods to soft/biological materials. The successful applicant will be expected to develop an externally funded research program, assist in the routine operation and management of the microscopy facilities, and have a commitment to cross-disciplinary education at both the undergraduate and graduate level. Candidates are expected to have a PhD in materials science, physics, chemistry or related engineering discipline.
Applicants should submit a letter of application, curriculum vitae (including list of publications), description of research and teaching plans, and names and contact information of at least three references online through the following link ( http://www.chms.ucdavis.edu/employment/recruitment/). The position is open until filled; but to assure full consideration, online applications should be submitted no later than November 30, 2006, for a targeted start date of July 1, 2007.
In all seriousness Hitachi has really done a good job with this little machine. We were invited to host a 2 day demo and I agreed, though with reservation considering the lack of options, control and detectors. From crate to first image was about 1.5-2 hours. This happened to be during our "bring your kids to work" day. I opened the lab up to the researchers and their families and the traffic was non-stop. After 10 minutes of instruction, my 9yr old son ran all the demos the first day on an astounding array of samples. It has extremely limited choice (often none) in terms of the usual Kv, spotsize gas pressure etc. but it produced publishable images of nearly every sample we threw at it from single celled dinoflagellates to crystal skulls, to bellybutton lint, almost all of it uncoated. At the end of the demo the only real negative thing anyone could say was that the stage did not tilt. I understand that has been rectified and that EDS capability has been added. While it currently cannot fully replace a dedicated research tool I estimate it could adequately image 75% or more of the biologic material our taxonomists study with virtually guaranteed results, no steep learning curve and minimal training. It required a simple wall outlet, and minimal bench space. I actually considered putting it out on the exhibit floor. We only had it two days but I was duly impressed. At the conclusion, my son was speaking to the engineers and called it the "easiest microscope in the world". Guess it's time to start looking for a new profession...
Scott Whittaker NMNH Imaging Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-633-0891
-----Original Message----- X-from: bozhilov-at-ucr.edu [mailto:bozhilov-at-ucr.edu] Sent: Tuesday, September 19, 2006 11:03 AM To: Whittaker, Scott
A feedback of any experience with the Hitachi TM1000 low vacuum desktop scanning electron microscope regarding resolution, performance, reliability, costs, etc. will be appreciated.
Thank you,
Krassimir N. Bozhilov Central Facility for Advanced Microscopy and Microanalysis University of California Riverside, CA 92521
tel 951 827 2998 fax 951 827 2489 bozhilov-at-ucr.edu
==============================Original Headers============================== 7, 19 -- From bozhilov-at-ucr.edu Tue Sep 19 10:01:12 2006 7, 19 -- Received: from sentry.ucr.edu (sentry.ucr.edu [138.23.226.224]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8JF1Acd011010 7, 19 -- for {microscopy-at-microscopy.com} ; Tue, 19 Sep 2006 10:01:11 -0500 7, 19 -- Received: from [138.23.185.162] (igppb116g.ucr.edu [138.23.185.162]) 7, 19 -- by sentry.ucr.edu (MOS 3.5.9-GR) 7, 19 -- with ESMTP id DZU42628 (AUTH via LOGINBEFORESMTP) 7, 19 -- for {microscopy-at-microscopy.com} ; 7, 19 -- Tue, 19 Sep 2006 08:01:04 -0700 (PDT) 7, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 7, 19 -- Content-Transfer-Encoding: 7bit 7, 19 -- Message-Id: {85F81D99-9C1B-4C52-A0F0-96C3A80051D4-at-ucr.edu} 7, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- From: KN Bozhilov {bozhilov-at-ucr.edu} 7, 19 -- Subject: Hitachi TM1000 7, 19 -- Date: Tue, 19 Sep 2006 08:01:04 -0700 7, 19 -- X-Mailer: Apple Mail (2.752.2) 7, 19 -- X-Junkmail-Status: score=0/65, host=sentry.ucr.edu ==============================End of - Headers==============================
==============================Original Headers============================== 16, 27 -- From WHITTAKS-at-si.edu Wed Sep 20 08:42:46 2006 16, 27 -- Received: from si-mailout02.si.edu (si-mailout02.si.edu [160.111.103.154]) 16, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8KDgjME013559 16, 27 -- for {microscopy-at-microscopy.com} ; Wed, 20 Sep 2006 08:42:46 -0500 16, 27 -- Received: from SI-MSESMTPO-02.US.SINET.SI.EDU (SI-MSESMTP-N2.us.sinet.si.edu [160.111.49.76]) 16, 27 -- by si-mailout02.si.edu (SI-Mailer) with ESMTP 16, 27 -- id A93C5D9E0; Wed, 20 Sep 2006 09:42:45 -0400 (EDT) 16, 27 -- Received: from SI-ECL02.US.SINET.SI.EDU ([160.111.49.26]) by SI-MSESMTPO-02.US.SINET.SI.EDU with Microsoft SMTPSVC(6.0.3790.1830); 16, 27 -- Wed, 20 Sep 2006 09:42:32 -0400 16, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 27 -- Content-class: urn:content-classes:message 16, 27 -- MIME-Version: 1.0 16, 27 -- Content-Type: text/plain; 16, 27 -- charset="US-ASCII" 16, 27 -- Subject: RE: [Microscopy] Hitachi TM1000 16, 27 -- Date: Wed, 20 Sep 2006 09:42:31 -0400 16, 27 -- Message-ID: {20CDD1CED2E76541A4FA119C6C806BA381F18B-at-SI-ECL02.US.SINET.SI.EDU} 16, 27 -- In-Reply-To: {200609191503.k8JF3Oic013878-at-ns.microscopy.com} 16, 27 -- X-MS-Has-Attach: 16, 27 -- X-MS-TNEF-Correlator: 16, 27 -- Thread-Topic: [Microscopy] Hitachi TM1000 16, 27 -- Thread-Index: Acbb/NDL2LCrlUgZTnCfJflcXcYi6wACECsw 16, 27 -- From: "Whittaker, Scott" {WHITTAKS-at-si.edu} 16, 27 -- To: {bozhilov-at-ucr.edu} 16, 27 -- X-OriginalArrivalTime: 20 Sep 2006 13:42:32.0573 (UTC) FILETIME=[9F90C2D0:01C6DCBA] 16, 27 -- Content-Transfer-Encoding: 8bit 16, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8KDgjME013559 ==============================End of - Headers==============================
Try angstrom.us (various brands) or semtech solutions (Amray).
gary g.
At 06:22 AM 9/20/2006, you wrote:
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Thanks All, for the multitude of responses to my query. The majority vote for a slight difference in tilt angle. It is no surprise to me that tilt/incident beam angle makes quite a difference. ...Ever try to assemble a low magnification mosaic from BSE grain channeling images? Impossible to match/stitch! In this case, I *thought* I was close enough to zero to minimize the problem, I guess not.
Apparently I cannot (easily) reproduce exactly zero degrees. I am very busy here, but if time permits will try again, making minute tilt changes and observing the result to confirm the effect. This will, however, not be easy with my crude stage tilt mechanism. The relatively new digital SEM stage tilt is not very precise or controllable; much worse than my good old ETEC was. :(
Woody White BWXT Services
==============================Original Headers============================== 5, 26 -- From nwwhite-at-bwxt.com Wed Sep 20 15:20:09 2006 5, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 5, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8KKK9RY009516 5, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 20 Sep 2006 15:20:09 -0500 5, 26 -- Received: from ([131.184.13.224]) 5, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.2241081; 5, 26 -- Wed, 20 Sep 2006 16:19:58 -0400 5, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 5, 26 -- Wed, 20 Sep 2006 16:19:58 -0400 5, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 26 -- Content-class: urn:content-classes:message 5, 26 -- MIME-Version: 1.0 5, 26 -- Content-Type: text/plain; 5, 26 -- charset="US-ASCII" 5, 26 -- Subject: BSE Image Response - Thanks! 5, 26 -- Date: Wed, 20 Sep 2006 16:19:58 -0400 5, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D070-at-BWXSPO01.BWXS.BWXTECH.NET} 5, 26 -- X-MS-Has-Attach: 5, 26 -- X-MS-TNEF-Correlator: 5, 26 -- Thread-Topic: BSE Image Response - Thanks! 5, 26 -- Thread-Index: Acbc8iSR3+lCxuc1TM2PDUzi6QiDEg== 5, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 5, 26 -- To: "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 5, 26 -- X-OriginalArrivalTime: 20 Sep 2006 20:19:58.0442 (UTC) FILETIME=[24CF9CA0:01C6DCF2] 5, 26 -- Content-Transfer-Encoding: 8bit 5, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8KKK9RY009516 ==============================End of - Headers==============================
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Title-Subject: [Filtered] Dispersion of polystyrene spheres
Question: I am trying to disperse a mixture of 5, 1, .5 micron polystyrene spheres to be used for calibration. I have tried several liquids, of which none have worked. Any help would be greatly appreciated. Thank you in advance.
Dear Listers, Please, reply me if you ever succeeded to realize Nanoindentation (especially local hardness evaluation) with Veeco SPM DI D3100. Thank you in advance, Best, Inna
Dr. Inna Popov Head of the Unit for Nanoscopic Characterization The Center for Nanoscience and Nanotechnology Faculty of Natural Science The Hebrew University of Jerusalem E. Safra Campus Givat Ram Jerusalem 91904 ISRAEL Phone: +972 2 6584808 Fax: +972 2 6584809 Email: innap-at-savion.huji.ac.il Web: www.nanoscience.huji.ac.il/unit
==============================Original Headers============================== 5, 26 -- From innap-at-savion.huji.ac.il Thu Sep 21 04:37:05 2006 5, 26 -- Received: from mail3.cc.huji.ac.il (real-outmail.cc.huji.ac.il [132.64.1.21]) 5, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8L9b4kq012023 5, 26 -- for {Microscopy-at-microscopy.com} ; Thu, 21 Sep 2006 04:37:04 -0500 5, 26 -- Received: from mango2.hustaff.huji.local (mango2.cc.huji.ac.il [132.64.16.16]) 5, 26 -- by mail3.cc.huji.ac.il (8.12.11.20060308/8.12.11) with ESMTP id k8L9b2D9009070 5, 26 -- for {Microscopy-at-microscopy.com} ; Thu, 21 Sep 2006 12:37:02 +0300 5, 26 -- Received: from MANGO1.hustaff.huji.local ([132.64.16.23]) by mango2.hustaff.huji.local with Microsoft SMTPSVC(6.0.3790.1830); 5, 26 -- Thu, 21 Sep 2006 12:37:02 +0300 5, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 26 -- Content-class: urn:content-classes:message 5, 26 -- MIME-Version: 1.0 5, 26 -- Content-Type: text/plain; 5, 26 -- charset="windows-1255" 5, 26 -- Subject: Nanoindentation on Veeco D3100 5, 26 -- Date: Thu, 21 Sep 2006 12:36:46 +0300 5, 26 -- Message-ID: {FCAF0955B591E644BEDEE450C0B58F4101155040-at-MANGO1.hustaff.huji.local} 5, 26 -- X-MS-Has-Attach: 5, 26 -- X-MS-TNEF-Correlator: 5, 26 -- Thread-Topic: Nanoindentation on Veeco D3100 5, 26 -- Thread-Index: AcbdYXSbwhvLo0qlSoyXuHhLos7iaA== 5, 26 -- From: "Inna Popov" {innap-at-savion.huji.ac.il} 5, 26 -- To: {Microscopy-at-microscopy.com} 5, 26 -- X-OriginalArrivalTime: 21 Sep 2006 09:37:02.0542 (UTC) FILETIME=[7E3226E0:01C6DD61] 5, 26 -- Content-Transfer-Encoding: 8bit 5, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8L9b4kq012023 ==============================End of - Headers==============================
Inna Popov asked about NanoIndentation using the NanoScope Dimension 3100 AFM sold by Veeco. We have done this and include this in our AFM analysis services. What would you like to know? 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: innap-at-savion.huji.ac.il To: donc-at-asmicro.com Sent: Thursday, September 21, 2006 5:40 AM Subject: [a] [Microscopy] Nanoindentation on Veeco D3100
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Dear Listers, Please, reply me if you ever succeeded to realize Nanoindentation (especially local hardness evaluation) with Veeco SPM DI D3100. Thank you in advance, Best, Inna
Dr. Inna Popov Head of the Unit for Nanoscopic Characterization The Center for Nanoscience and Nanotechnology Faculty of Natural Science The Hebrew University of Jerusalem E. Safra Campus Givat Ram Jerusalem 91904 ISRAEL Phone: +972 2 6584808 Fax: +972 2 6584809 Email: innap-at-savion.huji.ac.il Web: www.nanoscience.huji.ac.il/unit
==============================Original Headers============================== 16, 21 -- From donc-at-asmicro.com Thu Sep 21 17:01:21 2006 16, 21 -- Received: from smtp113.sbc.mail.re2.yahoo.com (smtp113.sbc.mail.re2.yahoo.com [68.142.229.92]) 16, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k8LM1LJM004199 16, 21 -- for {microscopy-at-microscopy.com} ; Thu, 21 Sep 2006 17:01:21 -0500 16, 21 -- Received: (qmail 26757 invoked from network); 21 Sep 2006 22:01:20 -0000 16, 21 -- Received: from unknown (HELO ASM11) (asmicro-at-sbcglobal.net-at-68.251.180.43 with login) 16, 21 -- by smtp113.sbc.mail.re2.yahoo.com with SMTP; 21 Sep 2006 22:01:20 -0000 16, 21 -- Message-ID: {000e01c6ddc9$3b613870$6501a8c0-at-ASM11} 16, 21 -- From: "Don Chernoff at ASM" {donc-at-asmicro.com} 16, 21 -- To: {innap-at-savion.huji.ac.il} , "Microscopy List" {microscopy-at-microscopy.com} 16, 21 -- References: {200609210940.k8L9ednC017479-at-ns.microscopy.com} 16, 21 -- Subject: [Microscopy] Nanoindentation on Veeco D3100 16, 21 -- Date: Thu, 21 Sep 2006 17:42:11 -0400 16, 21 -- MIME-Version: 1.0 16, 21 -- Content-Type: text/plain; 16, 21 -- charset="iso-8859-1" 16, 21 -- Content-Transfer-Encoding: 7bit 16, 21 -- X-Priority: 3 16, 21 -- X-MSMail-Priority: Normal 16, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1807 16, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 ==============================End of - Headers==============================
My question is not about microscopy, but I put my question on the list as a bottle in the sea !
I'm looking after the instructions manuel for an old pyrometer I've descoverd here, a KT4 from Heimann GmbH (Germany, now Heitronic). It's a nice infrared pyrometer, with a Cassegrain telescope as measuring head, which cover the range from -100 to +300°C.
Does some know this tool, and could send me a copie of the instructions manual ?
-- 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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Email: kca-at-soton.ac.uk Name: KC ANG
Organization: Postgraduate Student,University of Southampton
Title-Subject: [Filtered] Softwares to solve 3D measurements of SEM
Question: Hi,
I wish to know about the softwares that can solve and present 3D images for SEM. Currently, I only manage to have 2D images captured. Is it possible to measure angles of features using SEM?
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Email: andel-at-cperi.certh.gr Name: Andreas Delimitis
Organization: Centre for Research & Technology Hellas - CPERI
Title-Subject: [Filtered] CCD cameras for TEM
Question: Dear Listers,
I am looking for suggestions (vendors are also welcome - off list) for a bottom mounted CCD camera to be installed on a JEOL 2010 TEM. The microscope is primarily used for characterisation of electron beam sensitive materials, such as zeolites, catalysts and biological samples, so low-dose imaging conditions are required.
I am aware of two models from two major competitors in the market -but any other suggestion is more than welcome: the first provides a full-frame, 16 bit, 4.2 Mpx slow scan camera with a pixel size of 13.5x13.5 microns, while the second gives an interline, 14 bit, 11Mpx fast rate camera with a pixel size of 9x9 microns. However, there is no information about low-dose imaging, along with SAD, microdiffraction and CBED pattern aquisition for these cameras.
User suggestions are especially welcome.
Thanks in advance, Andreas
Dr. Andreas Delimitis Centre for Research & TEchnology Hellas (CERTH) Chemical Process Engineering Research Institute (CPERI) Thermi, Thessaloniki Greece
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Email: bpletc-at-ufl.edu Name: Ben Pletcher
Organization: University of Florida
Title-Subject: [Filtered] Tenupol-2 Sample Holder
Question: I have just relocated to the University of Florida and am repairing a Tenupol-2 electro-jet polisher. There are no sample holders. Struers does have a few left, but wanted to know if anyone had an old system lying around with spare sample holders and/or parts. I would greatly appreciate any help as I have a very limited budget. The parts of greatest help would be the light and sensor unit for automatically shut-off.
Anyone have an idea about how I can help a researcher interested in measuring microvolumes in his sample?
Here is the set up. Geologist laser drills holes into zircons and captures the 'smoke'. He does some fancy analysis and gets the info he needs, except, he wants to know the volume of the hole.
Holes are up to 40um in dia., between 5 and 20 um deep. Material is zircon, ZrSiO2.
He has used something called a vertical scanning interferometer, but the one he has used is broken or not longer available. He was looking for some other solution.
He came asking about confocal microscopy. I'm not sure our confocal will do anything for him, but we will give it a try. I suggested SEM, but he doesn't want to coat the sample. I thought FIB type machine, but we don't have one.
He says if he finds the right technique, he may be doing tens to hundreds of holes, so some automation or at least quick and easy procedure is what he is looking for.
Any ideas?
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 Sep 22 18:02:05 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 k8MN25WX032572 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 22 Sep 2006 18:02:05 -0500 12, 21 -- Received: from ucsc.edu (cruzmail-fe2.ucsc.edu [128.114.125.2]) 12, 21 -- by smtp-prod-mx1.ucsc.edu (8.13.7/8.13.7) with ESMTP id k8MN1Rke009217 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 22 Sep 2006 16:01:32 -0700 (PDT) 12, 21 -- Received: from [128.114.25.186] (account jmkrupp-at-ucsc.edu [128.114.25.186] verified) 12, 21 -- by silver.ucsc.edu (CommuniGate Pro SMTP 4.3.7) 12, 21 -- with ESMTPA id 71277760 for microscopy-at-microscopy.com; Fri, 22 Sep 2006 16:01:24 -0700 12, 21 -- Mime-Version: 1.0 12, 21 -- Message-Id: {p06230908c13a19609206-at-[128.114.25.186]} 12, 21 -- Date: Fri, 22 Sep 2006 16:01:43 -0700 12, 21 -- To: microscopy-at-microscopy.com 12, 21 -- From: Jon Krupp {jmkrupp-at-ucsc.edu} 12, 21 -- Subject: microvolume measuring 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==============================
This is easy with the Alicona Mex software. It allows "stereo" or even tri SEM images to be mathematically combined to produce a DEM. We use a VPSEM and there is no need to coat, one only needs a tilt stage.
john
At 04:06 PM 9/22/2006, you wrote:
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On Sep 22, 2006, at 4:02 PM, jmkrupp-at-ucsc.edu wrote:
} Anyone have an idea about how I can help a researcher interested in } measuring microvolumes in his sample? } } Here is the set up. Geologist laser drills holes into zircons and } captures the 'smoke'. He does some fancy analysis and gets the info } he needs, except, he wants to know the volume of the hole. } } Holes are up to 40um in dia., between 5 and 20 um deep. Material is } zircon, ZrSiO2. } } He has used something called a vertical scanning interferometer, but } the one he has used is broken or not longer available. He was looking } for some other solution. } } He came asking about confocal microscopy. I'm not sure our confocal } will do anything for him, but we will give it a try. I suggested SEM, } but he doesn't want to coat the sample. I thought FIB type machine, } but we don't have one. } } He says if he finds the right technique, he may be doing tens to } hundreds of holes, so some automation or at least quick and easy } procedure is what he is looking for. } } Any ideas? } Dear Jon, If the confocal can generate a reflection mode signal from the surface of the zircon, that should enable the estimation of the volumes of the holes. If, further, you know that the walls are not undercut, then the volumes will be measured pretty accurately. If the walls are undercut--the hole is wider at the bottom than at the top--you will have to tilt the specimen until the wall is visible all the way down, and it will take more than one tilt direction to see the whole wall, but the volume can be accurately measured. I have no good idea how to automate the process except in the case where the walls are either straight or converging, and where the holes are regularly spaced or can be found automatically, then a program such as Leginon could be adapted to collect confocal data. 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 Sep 22 18:50:57 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 (8.12.11.20060308/8.12.8) with ESMTP id k8MNov3L016394 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 22 Sep 2006 18:50:57 -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 DE700109FAF 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 22 Sep 2006 16:50:51 -0700 (PDT) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id CF336353A5 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 22 Sep 2006 16:50:50 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200609222302.k8MN2CPB032735-at-ns.microscopy.com} 4, 22 -- References: {200609222302.k8MN2CPB032735-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {d833e5b4c0e5f6dbe994b0629f98a522-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] microvolume measuring 4, 22 -- Date: Fri, 22 Sep 2006 16:54:26 -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 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 (equine.scientist-at-yahoo.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, September 24, 2006 at 12:32:49 ---------------------------------------------------------------------------
Email: equine.scientist-at-yahoo.com Name: Ryelei K. Sejavka
Organization: Kangaroo Valley Primary School
Education: K-8 Grade Grammar School
Location: Kangaroo Valley, New South Wales, Australia
Question: How do I find the manual for a really old microscope? On the front it says Sears 49-24364 400-1200 Power Z-O-O-M Microscope.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (keith-jackson-at-tesco.net) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, September 24, 2006 at 07:51:14 ---------------------------------------------------------------------------
Email: keith-jackson-at-tesco.net Name: Keith Jackson
Organization: Sandown Hiogh School
Education: 9-12th Grade High School
Location: Sandown, Isle of Wight, Hampshire, England
Question: How is the raw data picked up from the electrons processed to form the image that we see from a scanning electron microscope?
Diffusion processes in Mo-Si based alloys Research project: ----------------------- Better efficiency and low emissions of the future aircraft engines will require very high service temperature that the presently used nickel-based single crystal superalloys cannot sustain. Refractory metal silicide materials based on the Mo-Si system exhibit good properties in a temperature range up to 1300°C. A current research effort to enhance their oxidation resistance by alloying with diverse elements (B, Ti, Nb, Ge,…) and by developing alumina coatings. The coatings are complex in compositions and involve precious metals (Pt, Ru, etc...). In this context a deep knowledge of phase diagrams and diffusion mechanisms are required in order to optimize the different processes. The proposed work is included in a global effort to experimentally characterize phase diagrams and thermodynamical properties. A theoretical research focused at modeling the phase diagrams is also conducted in parallel. The post-doc researcher will be in charge of managing and analyzing diffusion couple experiments in order to obtain isothermal sections of ternary and quaternary diagrams of interest. This work will include sophisticated data analysis techniques and characterizations by analytical techniques (EPMA, SEM and TEM). A fundamental knowledge of diffusion mechanisms in these compounds at microscopic and intermediate scale is foreseen. Candidate The candidate has strong solid state physics background and must be well trained in computerized data analysis. He/She will work in strong collaboration with other experimentalists (thermodynamics measurements, crystallography, etc…) and theorists (ab initio and Calphad-Dictra). This project is part of a 3 years project funded by the ANR (French Research National Agency). Application: Application (letter of motivation + short CV with a list of publications + names and contact information for two references) should be sent to: Gilles.Hug-at-onera.fr
Dr. Gilles Hug LEM ONERA-CNRS BP 72 92322 Châtillon France +33 1 46 73 45 42 gilles.hug-at-onera.fr
==============================Original Headers============================== 6, 20 -- From gilles.hug-at-onera.fr Mon Sep 25 03:07:16 2006 6, 20 -- Received: from onera.onera.fr (onera.onera.fr [144.204.65.4]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8P87FrY029501 6, 20 -- for {microscopy-at-microscopy.com} ; Mon, 25 Sep 2006 03:07:16 -0500 6, 20 -- Received: from zig.onera.fr (localhost [127.0.0.1]) 6, 20 -- by onera.onera.fr with ESMTP id k8P87FDn026989 6, 20 -- for {microscopy-at-microscopy.com} ; Mon, 25 Sep 2006 10:07:15 +0200 (MEST) 6, 20 -- Received: from [195.221.211.30] (onthestep.onera.fr [195.221.211.30]) 6, 20 -- by zig.onera.fr (8.13.6+Sun/8.13.6/1.1) with ESMTP id k8P87E4l028089 6, 20 -- for {microscopy-at-microscopy.com} ; Mon, 25 Sep 2006 10:07:15 +0200 (CEST) 6, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 6, 20 -- Message-Id: {C6143A4F-1F6B-43F6-A383-2BC98AAE31D1-at-onera.fr} 6, 20 -- Content-Type: text/plain; charset=WINDOWS-1252; delsp=yes; format=flowed 6, 20 -- To: microscopy-at-microscopy.com 6, 20 -- From: Gilles Hug {gilles.hug-at-onera.fr} 6, 20 -- Subject: Post-doctoral fellowship at Onera (France) 6, 20 -- Date: Mon, 25 Sep 2006 10:07:14 +0200 6, 20 -- X-Mailer: Apple Mail (2.752.2) 6, 20 -- Content-Transfer-Encoding: 8bit 6, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8P87FrY029501 ==============================End of - Headers==============================
} Does anybody know about the electron speed of Kodak SO163 at 200 KV - } development in D19 full strength for 12 min.
The last time I measured this was back in 1996. I use a SS-CCD camera now... There have been formulation adjustments...
I measured:
Quantity KV 100 120 200 Unit density exposure [e/sq um] 0.67 0.81 1.10 (no correction for backscattering) Speed (sq um/e) 1.50 1.23 0.91 (no correction for backscattering)
Unit density exposure [e/sq um] 0.77 0.92 1.24 (with correction for backscattering) Speed (sq um/e) 1.30 1.08 0.81 (with correction for backscattering)
Note that my measured speed at 100 kV was 21% below that reported by Downing and Grano (Ultramicroscopy, 7, 381-404 1982). There had been formulation changes between in the interim...
To do careful work you really should measure it yourself...
Dear Keith, I have not seen an answer to your question, so I will attempt to explain. The electrons given off from the sample when it is imaged in the SEM are collected by the Secondary Electron (SE) Detector, because it has a positive charge that attracts the electrons. These SEs strike a fluorescent material on the inside end of the detector, which turns them into light, much like the material that coats the inside of the television glass. This light is amplified and turned into an electrical signal that modifies the viewing screen. As the electron beam scans over the sample surface, the viewing screen shows the variation in signal that is picked up by the SE detector at the same time. This builds up an image that looks a bit like what we see when we look at a sample with our own eyes, so we can understand it. There are lots of good Web sites that can illustrate this with nice pictures and animation, if you Google SEM. Good luck, Mary Mager Electron Microscopist Materials Eng. UBC #319 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA Phone: 604-822-5648 Fax: 604-822-3619 email: mager-at-interchange.ubc.ca -----Original Message----- X-from: keith-jackson-at-tesco.net [mailto:keith-jackson-at-tesco.net] Sent: Sunday, September 24, 2006 3:15 PM To: mager-at-interchange.ubc.ca
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (keith-jackson-at-tesco.net) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Sunday, September 24, 2006 at 07:51:14 ---------------------------------------------------------------------------
Email: keith-jackson-at-tesco.net Name: Keith Jackson
Organization: Sandown Hiogh School
Education: 9-12th Grade High School
Location: Sandown, Isle of Wight, Hampshire, England
Question: How is the raw data picked up from the electrons processed to form the image that we see from a scanning electron microscope?
I have a rather unusual request. I'm sending this out on the microscopy list server hoping it will result in useful feedback and knowing that it is not a subject of interest to the group as a whole. Therefore, I ask that any replies be sent directly to me.
If anyone would like the information garnered, I would be happy to share it.
Basically, I am looking for ideas, experiences and suggestions.
I would like to help set up a microscopy lab to be used primarily by my local high school as a teaching tool.
I envision a lab to also be used by a wider area than just one high school, so it would include making it available to other area high schools, technical schools and non-profit research groups. Those are what come to mind at the moment anyway.
Any and all thoughts, experiences and suggestions as to how to set up something like this, how to fund it, how to schedule it, what kinds of instrumentation would be best to have in it, where pitfalls may lie, etc. etc. are welcome. I would very much like to hear what you have to say.
Thank you,
dj
==============================Original Headers============================== 10, 16 -- From dljones-at-bestweb.net Tue Sep 26 09:49:10 2006 10, 16 -- Received: from vms044pub.verizon.net (vms044pub.verizon.net [206.46.252.44]) 10, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8QEnA1w020346 10, 16 -- for {Microscopy-at-microscopy.com} ; Tue, 26 Sep 2006 09:49:10 -0500 10, 16 -- Received: from localhost ([71.247.114.109]) 10, 16 -- by vms044.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep 10, 16 -- 9 2005)) with ESMTPA id {0J6700BRGFP0HL94-at-vms044.mailsrvcs.net} for 10, 16 -- Microscopy-at-microscopy.com; Tue, 26 Sep 2006 09:46:18 -0500 (CDT) 10, 16 -- Date: Tue, 26 Sep 2006 10:46:12 -0400 (Eastern Standard Time) 10, 16 -- From: "David L. Jones" {dljones-at-bestweb.net} 10, 16 -- Subject: Brain Hash 10, 16 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 10, 16 -- To: Microscopy-at-microscopy.com 10, 16 -- Message-id: {Pine.WNT.4.64.0609261044420.3164-at-H-F1} 10, 16 -- MIME-version: 1.0 10, 16 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed ==============================End of - Headers==============================
I am going to get my Robinson detector block recoated at a reputable coater here in Chicago. I need to know the thickness of the required coating in angstroms, which I understand is thinner than typical. If any of you have that info or any other special instructions I would appreciate it.
Thanks!
Tom Kaye
==============================Original Headers============================== 7, 20 -- From tom-at-tomkaye.com Tue Sep 26 15:35:50 2006 7, 20 -- Received: from tomkaye.com (mxgateway.techpro.com [64.4.207.8]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8QKZnE5000429 7, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 26 Sep 2006 15:35:49 -0500 7, 20 -- Received: from tkdell [24.15.58.103] by tomkaye.com with ESMTP 7, 20 -- (SMTPD32-8.14) id AF23610400F8; Tue, 26 Sep 2006 15:35:47 -0500 7, 20 -- From: "Tom Kaye" {tom-at-tomkaye.com} 7, 20 -- To: {Microscopy-at-microscopy.com} 7, 20 -- Subject: Robinson Detector coating thickness 7, 20 -- Date: Tue, 26 Sep 2006 15:35:45 -0500 7, 20 -- Message-ID: {003401c6e1ab$58412780$030a1aac-at-tkdell} 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="iso-8859-1" 7, 20 -- Content-Transfer-Encoding: 7bit 7, 20 -- X-Priority: 3 (Normal) 7, 20 -- X-MSMail-Priority: Normal 7, 20 -- X-Mailer: Microsoft Outlook CWS, Build 9.0.2416 (9.0.2910.0) 7, 20 -- Importance: Normal 7, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both john.green-at-uvm.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: john.green-at-uvm.edu Name: John Green
Organization: University of Vermont
Title-Subject: [Filtered] ventricle embedding problem
Question: I'm using glycol methacrylate (Technovit 7100) to embed rat forebrains for the purpose of stereology. However, sometimes, the ventricles fail to embed properly, even though the rest of the block cures perfectly. Thus, I sometimes get holes in the resin block, but only where the ventricles are. Does anyone have a solution to this problem? The brain tissue is fixed with 10% buffered formalin and then postfixed for several weeks at room temperature in the same solution. We then dehydrate it over the course of a day, gradually infiltrate it with the embedding solution over the course of a week, and then polymerize, all at room temperature. Infiltrations take place in light-sensitive vials rotated on a shaker.
} Date: Wed, 27 Sep 2006 09:52:55 +0100 } To: john.green-at-uvm.edu } From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} } Subject: Re: [Microscopy] viaWWW: ventricle embedding problem } Cc: Microscopy-at-microscopy.com } } Hi John, } } This seems like an infiltration problem. } } Check the block surface using a stereo microscope - if the problem area is } soft (prod with the points of forceps) that pretty well confirms it. } } You may find that incubating the affected blocks in a 50 - 60'C oven may } improve this - start with an incubation of 1hr and let the block cool down } before sectioning. If you think progress is being made increase the time } to overnight incubation and try again. } } To avoid this recurring you could maybe try out the following: } } 1) Trim the tissue to as thin a slice as possible. } } 2) Give increasing concentrations of EtOH/resin mixes of several hours or } overnight in say, 3:1, 1:1 and 1:3 dilutions prior to going to pure resin. } } 3) Slow down the polymerisation rate by using a crushed ice slush heat } sink surrounding the molds during polymerisation and/or reducing the } ammount of accelerator. } } Hope this helps, but get back to me if you want to discuss further. } } Alastair } At 17:54 26/09/2006 -0500, you wrote: } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Alastair McKinnon Histology & EM Facility Manager University of Aberdeen, Institute of Medical Science Foresterhill, Aberdeen, AB25 2ZD tel: +44(0) 1224 552923 (office); +44(0) 1224 555911 (lab) fax: +44(0) 1224 559533; email: a.d.mckinnon-at-abdn.ac.uk www.abdn.ac.uk/ims/h-em
==============================Original Headers============================== 5, 17 -- From a.d.mckinnon-at-abdn.ac.uk Wed Sep 27 03:54:58 2006 5, 17 -- Received: from mailhub3.abdn.ac.uk (mailhub3.abdn.ac.uk [139.133.7.13]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8R8swHW020348 5, 17 -- for {Microscopy-at-microscopy.com} ; Wed, 27 Sep 2006 03:54:58 -0500 5, 17 -- Received: from med-0069.ims.abdn.ac.uk ([139.133.159.90] helo=med-0069.abdn.ac.uk) 5, 17 -- by mailhub3.abdn.ac.uk with esmtp (Exim 4.52) 5, 17 -- id 1GSVBt-00080P-Nq; Wed, 27 Sep 2006 09:54:53 +0100 5, 17 -- Message-Id: {5.2.1.1.0.20060927095355.0110fb18-at-mailms.abdn.ac.uk} 5, 17 -- X-Sender: pat081-at-mailms.abdn.ac.uk 5, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 5, 17 -- Date: Wed, 27 Sep 2006 09:54:38 +0100 5, 17 -- To: john.green-at-uvm.edu 5, 17 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 5, 17 -- Subject: Fwd: Re: [Microscopy] viaWWW: ventricle embedding problem 5, 17 -- Cc: Microscopy-at-microscopy.com 5, 17 -- Mime-Version: 1.0 5, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
The FEI-Philips corporation has a great booklet explaining electron microscopy (TEM, SEM, (S)TEM): "Everything you wanted to know about EM but but didn't dare to ask". You can download the free PDF file at the following URL: http://www.fei.com/Resources/StudentLearning/tabid/91/Default.aspx
They also have a great listing of web resources for Teachers & Students at the following URL: http://www.fei.com/Resources/WebResourcesLinks/tabid/95/Default.aspx?s=1
Best regards,
Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"The ultimate human freedom is the ability to choose one's attitude in a given set of circumstances." -Viktor Frankl
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==============================Original Headers============================== 7, 29 -- From Walter.Bobrowski-at-pfizer.com Wed Sep 27 06:39:37 2006 7, 29 -- Received: from gromsgom01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 7, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8RBdYTA010238 7, 29 -- for {Microscopy-at-microscopy.com} ; Wed, 27 Sep 2006 06:39:37 -0500 7, 29 -- Received: from mopamrexc02.amer.pfizer.com (mopamrexc02.pfizer.com [170.116.30.68]) 7, 29 -- by gromsgom01.pfizer.com (8.13.4/8.13.4) with ESMTP id k8RBdKlW011670; 7, 29 -- Wed, 27 Sep 2006 07:39:20 -0400 7, 29 -- Received: from mopamrexc03.amer.pfizer.com ([170.116.30.69]) by mopamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 7, 29 -- Wed, 27 Sep 2006 07:39:34 -0400 7, 29 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc03.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 7, 29 -- Wed, 27 Sep 2006 07:39:34 -0400 7, 29 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.7226.0 7, 29 -- Content-class: urn:content-classes:message 7, 29 -- MIME-Version: 1.0 7, 29 -- Content-Type: text/plain; 7, 29 -- charset="US-ASCII" 7, 29 -- Subject: RE: AskAMicroscopist: SEM raw data & Imaging 7, 29 -- Date: Wed, 27 Sep 2006 07:39:32 -0400 7, 29 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D07ABF354-at-anaamrexm01.amer.pfizer.com} 7, 29 -- X-MS-Has-Attach: 7, 29 -- X-MS-TNEF-Correlator: 7, 29 -- Thread-Topic: RE: AskAMicroscopist: SEM raw data & Imaging 7, 29 -- Thread-Index: AcbhY6rxwVd/jUVYS1G+UcHofoRlyAAxb11g 7, 29 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 7, 29 -- To: {keith-jackson-at-tesco.net} , {Microscopy-at-microscopy.com} 7, 29 -- X-OriginalArrivalTime: 27 Sep 2006 11:39:34.0153 (UTC) FILETIME=[9A937790:01C6E229] 7, 29 -- X-Proofpoint-Spam-Reason: safe 7, 29 -- Content-Transfer-Encoding: 8bit 7, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8RBdYTA010238 ==============================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 pmoeck-at-pdx.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: pmoeck-at-pdx.edu Name: Peter Moeck
Organization: Portland State University
Title-Subject: [Filtered] Postdoc for up to 4 years, $35,500
Question: see www.physics.pdx.edu/~pmoeck/research group for links
Postdoc ñ (USA West coast based) ñ for a collaboration between Portland State University, the University of California at Davis, the University of Washington at Seattle, and the Pacific NorthWest National Laboratory
Initially for one year at $35,500 plus fringe benefits (health insurance, retirement benefits, etc.), available from January 2nd, 2007, onwards, extendable up to 4 years by mutual agreement.
A group of collaborators from Portland State University and the University of California at Davis, and the University of Washington at Seattle is seeking a (male/female) postdoc for a project on Crystallographic and spectroscopic analyses of ferromagnetic semiconducting nanoparticle aggregates with Curie temperatures well above room temperature.
A background in materials physics, materials chemistry, crystallography, or materials science and engineering is required. Familiarity with Z-contrast (HAADF) imaging in scanning transmission electron microscopes and associated electron energy loss spectroscopy are essential. Skills in high resolution phase-contrast transmission electron microscopy, electron diffraction and crystallography, and crystallographic image processing will be appreciated.
The search will be open until the position has been filled. Applications (CV, list of referees, reasons for coming to the US for applicants from aboard, list of publications if applicable, etc.) should be sent to any or all of the following collaborators:
Prof. Peter Moeck
Department of Physics
Portland State University
P.O. Box 751
Portland, Oregon 97207-0751
Tel.: 503 725 4227
Fax: 503 725 2815
e-mail: pmoeck-at-pdx.edu
research group: Nanocrystallography Group
free crystallographic on-line databases with approximately 10,000 structures
Prof. Nigel D. Browning
Department of Chemical Engineering and Materials Science University of California-Davis One Shields Ave Davis, CA 95616
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Email: pmoeck-at-pdx.edu Name: Peter Moeck
Organization: Portland State University
Title-Subject: [Filtered] postdoc or professional programmer with scientific background
Question: see www.physics.pdx.edu/~pmoeck/research group for links
Postdoc (or professional programmer with scientific background) ñ Portland State University (Portland, Oregon/USA)
Initially for one year at $35,000 (to $38,000 depending on experience and background) plus 50 % fringe benefits (health insurance, retirement benefits, etc.), available from January 2nd, 2007, onwards, extendable up to 3 years by mutual agreement and if continuing funding over ONAMI earmarks (or alternative funding agencies) can be secured.
The Nanocrystallography Group at the Department of Physics of Portland State University (PSU) is seeking a (male/female) postdoc (or professional programmer with scientific background) for a project on Image-based nanocrystallography with database support: Lattice-fringe fingerprinting to identify unknown nanocrystal phases & Nanocrystal morphology from tilt protocols.
Excellent computer skills (e.g. C++, JScript, Java, Python, Perl, PHP, and MySQL) are essential. A background in crystallography, mineralogy, or materials science and engineering at the Masters (or PhD level) is desirable. Advanced degrees in computer science may substitute.
PSU is with an estimated enrollment of about 30,000 students Oregonís largest and only urban university. It is located in downtown Portland/Oregon, i.e. the metropolis of the so called ìsilicon forestî. Since high tech industries blend in with abundant natural beauty in this region and because of its moderate climate, Portland has consistently been ranked in the top ten of ìAmericaís most livable citiesî.
The search will be open until the position has been filled. Applications (CV, list of referees, reasons for coming to the US for applicants from aboard, list of publications if applicable, etc.) should be sent to:
Prof. Peter Moeck
Department of Physics
Portland State University
P.O. Box 751
Portland, Oregon 97207-0751
http://www.physics.pdx.edu/~pmoeck
Applications by email to pmoeck-at-pdx.edu with attachments as *.pdf files are acceptable. Since there is a hyperlink at the title of the project above, prospective candidates are advised to deliver a statement on how they might contribute to this multi-institutional collaborative effort.
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Email: pmoeck-at-pdx.edu Name: Peter Moeck
Organization: Portland State University
Title-Subject: [Filtered] opportunity for graduate students
Question: see http://www.physics.pdx.edu/~pmoeck/research%20group/ for links
Graduate Students ñ Department of Physics, Portland State University (Oregon/USA)
MSc/PhD candidate positions in crystallography/applied physics
The Nanocrystallography Group at the Department of Physics of Portland State University is seeking candidates (m/f) for MSc and PhD projects in the field of transmission electron microscopy (TEM) based nanocrystallography. The graduate student projects focus on aspects of the development of novel nanocrystal characterization methods. These methods will utilize a combination of high-resolution phase contrast imaging in TEM and goniometry of direct lattice vectors.
The Nanocrystallography group is a user of Portland State Universityís electron microscopy center. Occasional travel to and work at the National Center for Electron Microscopy, Lawrence Berkeley National Laboratory is anticipated to access some of the most advanced TEM instrumentation in the country. Summer research at the Technical University Chemnitz, Germany, may also become part of some projects.
The ideal candidate has a diploma of BSc. in crystallography, mineralogy, materials science and engineering, physics, or chemistry, and is interested in both geometrical-structural crystallography and transmission electron microscopy. Computer- and experimental skills are essential for the success of the projects. While some projects will require more experimental skills, some other projects will require more computer skills (notably JScript, Java, C++, Perl, PHP).
Portland State University is with an estimated enrollment of about 30,000 students Oregonís largest and only urban university. Portland/Oregon is the major city in the so called ìsilicon forestî, i.e. a region where high tech industries blend in with abundant natural beauty, and has consistently been ranked in the top ten of ìAmericaís most livable citiesî. Information on student life in Portland can be found on our departmental web page. The search will be open until successful candidates have taken up their respective positions.
Applications (CV, references, reasons for coming to the USA, papers if applicable, etc.) should be sent to:
I was wondering if anyone on the board had any advice for venting electron microscope columns/sample chambers. In particular, we want a system that allows the column or sample chamber to reach atmospheric pressure by venting with dry N2 but does not allow the chamber to over pressurize above atmosphere to protect seals, thin window on EDS detectors, etc.
I’m aware that there are on-demand gas regulators that can control this sort of thing. Does anyone have any suggestions for a good vendor or model number for this sort of regulator?
Alternatively, I’ve also heard that demand valves that divers use are good for this sort of application as it only supplies gas when the diver breathes or sucks through the mouthpiece (or in this case when the chamber is still under vacuum). Does anyone know a particular brand or type of diver demand valve that works well for this application and is easy to modify (i.e. relatively easy to install standard fittings on the ends like NPT, etc.)?
Any comments or suggestions would be greatly appreciated. Thanks in advance,
preston
-- Preston Larson Research Scientist Samuel Roberts Noble Electron Microscopy Laboratory 770 Van Vleet Oval Phone: (405) 325-4391
==============================Original Headers============================== 9, 17 -- From plarson-at-ou.edu Wed Sep 27 10:29:15 2006 9, 17 -- Received: from c3p0.ou.edu (mail.zero.ou.edu [129.15.0.75]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8RFTFwv029604 9, 17 -- for {Microscopy-at-Microscopy.Com} ; Wed, 27 Sep 2006 10:29:15 -0500 9, 17 -- Received: from [127.0.0.1] ([129.15.39.56]) 9, 17 -- by c3p0.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 9, 17 -- 2005)) with ESMTP id {0J690035JCCPEI80-at-c3p0.ou.edu} for 9, 17 -- Microscopy-at-Microscopy.Com; Wed, 27 Sep 2006 10:29:13 -0500 (CDT) 9, 17 -- Date: Wed, 27 Sep 2006 10:29:11 -0500 9, 17 -- From: Preston Larson {plarson-at-ou.edu} 9, 17 -- Subject: Venting EM Chambers 9, 17 -- To: Microscopy-at-Microscopy.Com 9, 17 -- Message-id: {451A98C7.6000009-at-ou.edu} 9, 17 -- MIME-version: 1.0 9, 17 -- Content-type: text/plain; charset=windows-1252; format=flowed 9, 17 -- Content-transfer-encoding: 8BIT 9, 17 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) ==============================End of - Headers==============================
We use helium with our Hitachi VP-SEM for both the residual atmosphere and venting. We set the regulator to less that 1 psi which is enough to vent the chamber in about a minute. Of course, someone could inadvertently change the regulator setting.
Therefore, we also have a commercial (Circle Seal), spring-loaded pop-off valve in the line from the tank. It releases at 2 psi, if I recall correctly. It effectively guarantees we won't over-pressure any more than that. We have a similar valve tapped into the EDS mounting plate on our JEOL. I don't remember if that was installed with our EDS system by the EDS people or if it was an afterthought. The pop-off valves look like they should cost only about $5. They probably cost more, but they have got to be a lot cheaper than a on-demand regulator.
Also, we have unbolted the front of both microscope chambers. The Hitachi only loads through the front of the chamber. The JEOL has a load-lock, but we sometimes have to open the chamber for big samples. That way, any extra pressure simply leaks out through the front of the chamber.
Warren
-----Original Message----- X-from: plarson-at-ou.edu [mailto:plarson-at-ou.edu] Sent: Wednesday, September 27, 2006 10:30 AM To: wesaia-at-iastate.edu
A simple single stage regulator will do the job. I use cylinder N2 the regulator to drop tank pressure from 1800psi to 8psi. This then goes through a molecular sieve to dry the N2, then into the SEM chamber through the vent valve. Depending on the size of your chamber, just wait until the door slides open. If the SEM is set up correctly, it takes very little force to move the door open when completely vented.
If you have any ion pumps, be sure to only use N2, or air if no N2.
gary g.
At 08:31 AM 9/27/2006, you wrote:
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I made a demand regulator from a scuba regulator and it works well.
On the high pressure side of the scuba regulator, I cut the pressure hose and used a 1/4MPT - 1/4 brass hose barb adapter to attach the scuba regulator to the standard gas regulator on the nitrogen cylinder.
I got some relatively thick wall PVC tubing and slipped it over the mouthpiece using a hoseclamp as a retainer. To reduce the tube diameter for the microscope, I went to my local hardware store and got copper plumbing fittings. I just soldered together a number of pieces of copper pipe and reducing unions until I could switch to a brass pipe thread. The largest copper pipe fits snugly in the PVC tubing and I again used a hose clamp as a retainer.
I've been using several of these for backfilling TEM and SEM chambers for over 10 years. No launching TEM windows into the users' laps, no blown EDS detectors, etc.
Cheers, Henk
At 11:31 AM 09/27/06, plarson-at-ou.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.
==============================Original Headers============================== 12, 26 -- From colijn.1-at-osu.edu Wed Sep 27 12:49:51 2006 12, 26 -- Received: from ER6S1.ENG.OHIO-STATE.EDU (er6s1.ecr6.ohio-state.edu [164.107.76.2]) 12, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8RHnoNs031214 12, 26 -- for {microscopy-at-microscopy.com} ; Wed, 27 Sep 2006 12:49:51 -0500 12, 26 -- Received: from CONVERSION-DAEMON.er6s1.eng.ohio-state.edu by 12, 26 -- er6s1.eng.ohio-state.edu (PMDF V6.2-1x11 #31056) 12, 26 -- id {01M7OX1UBEOWA7BUKV-at-er6s1.eng.ohio-state.edu} for 12, 26 -- microscopy-at-microscopy.com; Wed, 27 Sep 2006 13:49:49 -0400 (EDT) 12, 26 -- Received: from HOC1.ecr6.ohio-state.edu 12, 26 -- (hoc1.ceof.ohio-state.edu [164.107.76.179]) by er6s1.eng.ohio-state.edu 12, 26 -- (PMDF V6.2-1x11 #31056) 12, 26 -- with ESMTPA id {01M7OX1T7JPMA8KKWP-at-er6s1.eng.ohio-state.edu} ; Wed, 12, 26 -- 27 Sep 2006 13:49:47 -0400 (EDT) 12, 26 -- Date: Wed, 27 Sep 2006 13:51:35 -0400 12, 26 -- From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} 12, 26 -- Subject: Re: [Microscopy] Venting EM Chambers 12, 26 -- In-reply-to: {200609271531.k8RFVWuc032433-at-ns.microscopy.com} 12, 26 -- Sender: colijn-at-er6s1.ecr6.ohio-state.edu 12, 26 -- To: plarson-at-ou.edu 12, 26 -- Cc: microscopy-at-microscopy.com 12, 26 -- Message-id: {7.0.1.0.2.20060927133902.03727660-at-osu.edu} 12, 26 -- MIME-version: 1.0 12, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 26 -- Content-type: text/plain; charset=us-ascii; format=flowed 12, 26 -- X-Env-From: auth/colijn.1-at-osu.edu 12, 26 -- References: {200609271531.k8RFVWuc032433-at-ns.microscopy.com} ==============================End of - Headers==============================
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Email: kunli218-at-yahoo.com.sg Name: Simon Lee
Organization: Chartered Semiconductor
Title-Subject: [Filtered] Software for accurate diffraction spot location identification
Question: Dear Listers,
We are doing some strain/stress analysis work using electron nano diffraction technique. But accurate identification of diffraction spot location is the challenge we are facing. Is there any software available for this purpose?
Imaging analyst position is available in the Molecular Imaging Unit (MIU, www.miu.helsinki.fi) at Biomedicum Helsinki, University of Helsinki.
MIU:
MIU is located at the dynamic and international environment of Biomedicum Helsinki, University of Helsinki. At the moment, the unit has approximately 150 users. The unit contains state-of-the-art instruments including Cellomics ArrayScan high-content screening platform, Zeiss LSM510 META laser scanning confocal microscope, Zeiss/3i Stallion live-cell imaging system, PerkinElmer UltraView spinning-disk confocal microscope, and epifluorescence research microscopes. Imaging software includes MediaCybernetics ImageProPlus and AutoQuant deconvolution software.
Job description:
You will work as a member of our imaging team. Your task will be to take care of microscope instruments, including the computers and imaging software, maintenance of MIU web pages, including an online reservation system, as well as user training, communication, troubleshooting, and documentation. You will also participate in teaching. Training for the duties will be provided.
Requirements:
M.Sc.Tech./M.Sc./B.Sc., laboratory engineer/engineer degree or equivalent. Ability to communicate in English is essential in our multinational environment. Computer skills are highly desired. Experience and training in microscopy and/or programming is also appreciated. The position will be available in November 2006. For further information, please contact MIU Imaging Coordinator Anne Vaahtokari, tel. +358-9-191-25579, email anne.vaahtokari-at-helsinki.fi.
The universities are implementing a new salary system, where the salaries will have both job-based and personal work performance-based components. The job-specific component of the salary is based on general staff level 7, which corresponds to 1843,45 € per month. Personal work performance component will be, at most, 46% of the job-specific component.
Applications should include: a letter stating your interests; possible previous experience in the use of microscopes and imaging software; computer skills; possible salary request; CV. Send your application to Dr. Anne Vaahtokari, either by email, or mail it to: Molecular Imaging Unit, Biomedicum Helsinki, University of Helsinki, PO BOX 63, FIN-00014 Helsinki, Finland. The deadline for applications is October 20, 2006.
-- Ville-Veikko Oksa, M.Sc. | Molecular Imaging Unit Imaging Analyst | Biomedicum Helsinki, Room B505b ville-veikko.oksa-at-helsinki.fi | PO Box 63 (Haartmaninkatu 8) Tel: +358-9-191-25579 (work) | FI-00014 University of Helsinki +358-40-744-3232 (mobile) | http://www.miu.helsinki.fi/
==============================Original Headers============================== 15, 28 -- From vvoksa-at-mappi.helsinki.fi Thu Sep 28 02:12:27 2006 15, 28 -- Received: from sender-02.it.helsinki.fi (sender-02.it.helsinki.fi [128.214.205.137]) 15, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8S7CPxQ000669 15, 28 -- for {Microscopy-at-microscopy.com} ; Thu, 28 Sep 2006 02:12:26 -0500 15, 28 -- Received: from quas.it.helsinki.fi (quas.it.helsinki.fi [128.214.205.29]) 15, 28 -- by sender-02.it.helsinki.fi (8.13.7/8.13.7) with ESMTP id k8S7CMQA000820 15, 28 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT); 15, 28 -- Thu, 28 Sep 2006 10:12:23 +0300 15, 28 -- Received: from quas.it.helsinki.fi (localhost.localdomain [127.0.0.1]) 15, 28 -- by quas.it.helsinki.fi (8.13.1/8.13.1) with ESMTP id k8S7B7Y5030577; 15, 28 -- Thu, 28 Sep 2006 10:11:07 +0300 15, 28 -- Received: (from apache-at-localhost) 15, 28 -- by quas.it.helsinki.fi (8.13.1/8.13.1/Submit) id k8S7B7KH030574; 15, 28 -- Thu, 28 Sep 2006 10:11:07 +0300 15, 28 -- Received: from 128.214.167.37 ( [128.214.167.37]) 15, 28 -- as user vvoksa-at-localhost by www1.helsinki.fi with HTTP; 15, 28 -- Thu, 28 Sep 2006 10:11:07 +0300 15, 28 -- Message-ID: {1159427467.451b758b3c840-at-www1.helsinki.fi} 15, 28 -- Date: Thu, 28 Sep 2006 10:11:07 +0300 15, 28 -- From: Ville-Veikko Oksa {ville-veikko.oksa-at-helsinki.fi} 15, 28 -- To: Microscopy-at-microscopy.com, CONFOCAL-at-LISTSERV.BUFFALO.EDU 15, 28 -- Subject: Position available - Imaging Analyst 15, 28 -- MIME-Version: 1.0 15, 28 -- Content-Type: text/plain; charset=ISO-8859-1 15, 28 -- Content-Transfer-Encoding: 8bit 15, 28 -- User-Agent: Internet Messaging Program (IMP) 3.1 15, 28 -- X-Originating-IP: 128.214.167.37 15, 28 -- Sender: vvoksa-at-mappi.helsinki.fi ==============================End of - Headers==============================
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Title-Subject: [Filtered] Looking for Used Ultramicrotome
Question: Greetings to all ~
My LKB-Huxley ultramicrotome is giving up the ghost; twice it has quit on me for no apparent reason, then starts up again (thankfully!) just as mysteriously. Spare or replacement parts are no longer available. The budget committee has already denied my request for a new ultramicrotome for the coming fiscal year, so I really need to find a quality used machine. Any ideas?
Thanks in advance - Rita Kenner HTL(ASCP) Electron Microscopist Marshfield Clinic Laboratories Marshfield, Wisconsin kenner.rita-at-marshfieldclinic.org (715)387-9159 -or- 1-800-782-8581, ext.79159
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Organization: Department of Physics, University of Calabria, Italy
Title-Subject: [Filtered] PHD position
Question: PhD position on nanostructured carbon materials at the University of Calabria, Italy
Applications are invited for a PhD student position in the Research Group "Surface electronic spectroscopies" (SPES) at the Department of Physics, University of Calabria, Arcavacata di Rende, Cosenza, Italy, (http://www.fis.unical.it/index.php?lingua=_eng), to begin December 2006.
The interests of the SPES group are in condensed matter and surface physics. In those fields the group leads his research in the determination of the chemical-physical properties of surfaces and solids mainly through the study of their electronic properties. The determination of the interaction between light molecules and surfaces and their peculiar electronic aspects is carried out by electronic spectroscopies. Many subjects are developed in the SPES group and, in particular, recently we focus on the characterization of nanostructured carbon materials for application in fuel cells, biosensors, supports for catalysis etc. The addition of metal clusters in the carbon matrix will allow a better control of the nano- and meso-structures and of the overall reactivity. The available experimental techniques are the EELS-TEM spectro-microscopy, SEM-EDS, photoemission spectroscopies (XPS, UPS), Auger spectroscopy, micro-Raman and UV-VIS-IR spectroscopy. Complementary results on the nano- and meso-structure can be obtained by TEM and SEM images. Finally the water sensitivity of functionalized C-materials will be investigated analysing the valence and conduction bands (by home-based and Synchrotron measurements, respectively) while the morphology changes will be monitored by SEM microscopy. Experience with vacuum system and a basic knowledge of solid state physics would be an asset for applicants. SPES provides an open and stimulating atmosphere and many opportunities to interact with other researchers are provided for by the PhD funds of the Physics Department at the University of Calabria. Interested applicants should contact Prof. Raffaele G. Agostino at agostino-at-fis.unical.it, tel. +39 0984 496162 / 496175 or Dr. Enrico Maccallini at maccallini-at-fis.unical.it, tel +39 0984 496107 / 496175.
I've been asked several times the following question:
How many high schools in the world actually have equipment such as SEM's or other fairly sophisticated scientific equipment for use in the classroom?
If you are a high school, or know of a high school that has this kind of capability, can you please post that information. I would like to know where you are, contact information, and what equipment you use.
I think it will be of interest for our whole scientific community to know the answer to that question, so I suggest that you post the answer to the group as a whole.
If you prefer to answer me directly, please indicate if you prefer information to be shared or not as the case may be.
Thank you,
dj
==============================Original Headers============================== 10, 16 -- From dljones-at-bestweb.net Thu Sep 28 09:15:24 2006 10, 16 -- Received: from vms044pub.verizon.net (vms044pub.verizon.net [206.46.252.44]) 10, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8SEFONA006752 10, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 28 Sep 2006 09:15:24 -0500 10, 16 -- Received: from localhost ([71.247.114.109]) 10, 16 -- by vms044.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep 10, 16 -- 9 2005)) with ESMTPA id {0J6B00G7F3LEFSQ0-at-vms044.mailsrvcs.net} for 10, 16 -- Microscopy-at-microscopy.com; Thu, 28 Sep 2006 09:15:18 -0500 (CDT) 10, 16 -- Date: Thu, 28 Sep 2006 10:27:44 -0400 (Eastern Standard Time) 10, 16 -- From: "David L. Jones" {dljones-at-bestweb.net} 10, 16 -- Subject: Perspectives: High School Microscopy 10, 16 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 10, 16 -- To: Microscopy-at-microscopy.com 10, 16 -- Message-id: {Pine.WNT.4.64.0609281008410.3292-at-H-F1} 10, 16 -- MIME-version: 1.0 10, 16 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed ==============================End of - Headers==============================
I'm going to perform my first immunogold labeling experiment. Could anybody provide an effective protocol for SEM immunogold labeling of proteins immobilized on surfaces? I don't have an histological slice, but only proteins adsorbed on a surface: do I need pre-fixation? Wich procedure is the best for detecting Human Serum ALBUMIN and not having any interference with the blocking solution?
Thanks for any suggestions Mariangela
Mariangela Fedel Dept of Materials Engineering and Industrial Technologies University of Trento Via Mesiano 77, 38100 Trento, Italy E-mail: mariangela.fedel-at-ing.unitn.it
==============================Original Headers============================== 6, 27 -- From Mariangela.Fedel-at-apss.tn.it Thu Sep 28 10:44:49 2006 6, 27 -- Received: from UITASMCAFEE2.apss.tn.it (uitasmcafee2.apss.tn.it [194.105.50.199]) 6, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k8SFimgf019118 6, 27 -- for {Microscopy-at-Microscopy.com} ; Thu, 28 Sep 2006 10:44:49 -0500 6, 27 -- Received: from (mail.apss.tn.it [194.105.50.212]) by UITASMCAFEE2.apss.tn.it with smtp 6, 27 -- id 3ef5_4519c462_4f08_11db_9b78_001143d31585; 6, 27 -- Thu, 28 Sep 2006 17:44:47 +0200 6, 27 -- Received: from w3gen06.apss.tn.it ([172.21.200.11]) by W3ITMAIL.apss.tn.it with Microsoft SMTPSVC(6.0.3790.1830); 6, 27 -- Thu, 28 Sep 2006 17:44:47 +0200 6, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 27 -- Content-class: urn:content-classes:message 6, 27 -- MIME-Version: 1.0 6, 27 -- Content-Type: text/plain; 6, 27 -- charset="us-ascii" 6, 27 -- Disposition-Notification-To: "Fedel Mariangela" {Mariangela.Fedel-at-apss.tn.it} 6, 27 -- Subject: SEM Immunogold labeling & Albumin 6, 27 -- Date: Thu, 28 Sep 2006 17:44:46 +0200 6, 27 -- Message-ID: {6633FFCE630D694D8FD6C310A5857A9F01C90C10-at-w3gen06.apss.tn.it} 6, 27 -- X-MS-Has-Attach: 6, 27 -- X-MS-TNEF-Correlator: 6, 27 -- Thread-Topic: SEM Immunogold labeling & Albumin 6, 27 -- Thread-Index: AcbjFQZER9sa3atTScudNMBKZjsQYw== 6, 27 -- From: "Fedel Mariangela" {Mariangela.Fedel-at-apss.tn.it} 6, 27 -- To: {Microscopy-at-Microscopy.com} 6, 27 -- X-OriginalArrivalTime: 28 Sep 2006 15:44:47.0278 (UTC) FILETIME=[06B1ECE0:01C6E315] 6, 27 -- Content-Transfer-Encoding: 8bit 6, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8SFimgf019118 ==============================End of - Headers==============================
Dear All, concerning your question: I organized and maintain SEMs and TEMs at two high-schools in Germany:
- at the Wilhelm-Loehe-Schule Nuerberg a JEOL 35C and a ZEISS EM9 S2 completely with ISI sputtercoater and LKB ultramicrotome
- at the Armin-Knab-Gymnasium Kitzingen a JEOL 840, which is still work in progress because the turbo failed start-up :-(( but hopefully will work end of October... Here I am looking for some lab equipment (coater...)
I am doing electron microcopy as a hobby since more than twenty years and I find it very rewarding to open up a new sight on structures (and life) with these instruments for pupils (and me).
So if you are thinking to give a SEM to a high-school: do it!
----- Original Message ----- X-from: {dljones-at-bestweb.net} To: {stefan.diller-at-t-online.de} Sent: Thursday, September 28, 2006 4:20 PM
Hi everybody,
I just wanted to thank everyone for the comments and suggestions regarding methods for not over pressurizing EM chambers.
From the responses, it sounds like on demand gas regulators might be a bit of overkill for this application and it looks like there are a number of much cheaper alternatives out there that I will begin to explore.
Once again, thanks to everyone who responded. It was greatly appreciated. preston
-- Preston Larson Research Scientist Samuel Roberts Noble Electron Microscopy Laboratory 770 Van Vleet Oval Phone: (405) 325-4391
==============================Original Headers============================== 7, 19 -- From plarson-at-ou.edu Thu Sep 28 12:31:24 2006 7, 19 -- Received: from r2d2.ou.edu (mail.zero.ou.edu [129.15.0.75]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8SHVOZn009734 7, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 28 Sep 2006 12:31:24 -0500 7, 19 -- Received: from [127.0.0.1] ([129.15.39.56]) 7, 19 -- by r2d2.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 7, 19 -- 2005)) with ESMTP id {0J6B00DM9CO8VV10-at-r2d2.ou.edu} for 7, 19 -- Microscopy-at-microscopy.com; Thu, 28 Sep 2006 12:31:22 -0500 (CDT) 7, 19 -- Date: Thu, 28 Sep 2006 12:31:22 -0500 7, 19 -- From: Preston Larson {plarson-at-ou.edu} 7, 19 -- Subject: Re: [Microscopy] Venting EM Chambers 7, 19 -- In-reply-to: {D17B4C65AAF187419DD2196BAC41C2AD01A6F8-at-exchange04.oxynet.oxy.edu} 7, 19 -- To: Microscopy-at-microscopy.com 7, 19 -- Message-id: {451C06EA.2000802-at-ou.edu} 7, 19 -- MIME-version: 1.0 7, 19 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 7, 19 -- Content-transfer-encoding: 7BIT 7, 19 -- References: {D17B4C65AAF187419DD2196BAC41C2AD01A6F8-at-exchange04.oxynet.oxy.edu} 7, 19 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) ==============================End of - Headers==============================
} I just wanted to thank everyone for the comments and suggestions } regarding methods for not over pressurizing EM chambers. } } From the responses, it sounds like on demand gas regulators might be a } bit of overkill for this application and it looks like there are a } number of much cheaper alternatives out there that I will begin to explore. } } Once again, thanks to everyone who responded. It was greatly appreciated. } preston
A final comment, If you have an EDS detector on the instrument or plan to install one in the future, you really need to get an on-demand regulator. An ultra-thin window will not survive much over pressure and the risk is too high. You risk a many thousand dollar repair bill by saving a hundred or so dollars. For reference check into a Matheson Model 3491, about $250 retail, ebay has one for $119.
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
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Perhaps I didn't note in my initial post about simple pop-off valves that both of our SEMs have light-element EDS detectors. They have been mounted for 10 years or more without any over-pressure incidents. I think a stray, flyaway particle may have punctured one of the panes on one detector necessitating a repair. However, it was a pinhole defect, not a big rip.
Detectors are a big investment and you want to make sure some protection system is in place and that it is not going to fail. Still, cheap solutions are available and might be more fail-safe than more expensive and complicated systems.
Warren Straszheim Materials Analysis and Research Laboratory Iowa State University
X-from: davilla-at-4pi.com [mailto:davilla-at-4pi.com] Sent: Thu 9/28/2006 1:04 PM To: wesaia-at-iastate.edu
Dear Mariangela,
Your setup is a little bit comparable to labelling of viruses on a grid film surface. But it is not an easy one since your target is so much smaller. However there are a few simple tests that may at least help you get started. Is pre-fixation necessary? Maybe not, provided the target protein will stick to the surface during the specimen handling (immuno incubations). You could check this in a 'dry run'. It is tempting to suggest a poly-L-lysine coat on the surface to potentially make the proteins stick more tightly, but this may cause some background with immunogold, especially when you are limited in the choice of blocking and incubation media, as you mentioned. So I would start of with a clean surface and protein adsorbed onto it and check whether it will stick during the incubations.
How to proceed for the detection of human albumin and achieve acceptable signal to noise ratios? Using bovine albumin in a blocking solution may be a problem if there are epitopes common to both BSA and HSA. The same holds for using any normal sera, as they contain albumins as well. If that info is not available that can be checked e.g. using a dot spot test. I do not have a reference at hand, but acetylated BSA (BSAc™) seemed not to interfere with the detection of albumins. I can find that info if you like.
Is using a protein block and incubation medium additive going to interfere with what you will see on the specimen carrier surface? It is very likely that some areas that were not previously covered with HSA will be so by components from a protein blocking solution or incubation medium. Will that be a problem in terms of visualizing the HSA as isolated molecules?
Is there an alternative? An incubation based on a detergent compound at a concentration slightly above the critical micelle concentration (in practice Tween-20 at 0.1% or alternatively Triton X-100 at 0.05%) might theoretically do the trick, but there is a fair chance that the adsorbed proteins will get washed away. Again this is something that could be checked in a dry run. Washing off can probably be largely prevented by using a pre-fixation after all, but again, this has its own limitations with risk of loss of recognition, increased background as well as irregular, cloggy surfaces.
Your questions raise many others, but with the indicated simple tests you should start getting some ideas.
Please feel free to get in touch if you would like to discuss the matter further.
Good luck
Jan Leunissen
Aurion - President Electron Microscopy Research Advisor Costerweg 5 Dept Anatomy and Structural Biology 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4795465 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz -----------------------------------------------------------------
==============================Original Headers============================== 13, 21 -- From leunissen-at-aurion.nl Thu Sep 28 16:16:15 2006 13, 21 -- Received: from fep04.xtra.co.nz (fep04.xtra.co.nz [210.54.141.242]) 13, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8SLGEJW019859 13, 21 -- for {Microscopy-at-microscopy.com} ; Thu, 28 Sep 2006 16:16:14 -0500 13, 21 -- Received: from [192.168.1.50] (really [222.152.168.157]) 13, 21 -- by fep04.xtra.co.nz with ESMTP 13, 21 -- id {20060928211609.PXQD11710.fep04.xtra.co.nz-at-[192.168.1.50]} ; 13, 21 -- Fri, 29 Sep 2006 09:16:09 +1200 13, 21 -- In-Reply-To: {200609281545.k8SFjHcg019585-at-ns.microscopy.com} 13, 21 -- References: {200609281545.k8SFjHcg019585-at-ns.microscopy.com} 13, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 13, 21 -- Content-Type: text/plain; charset=WINDOWS-1252; delsp=yes; format=flowed 13, 21 -- Message-Id: {CD8DDB18-43BD-4683-8639-51D0AF1ED33E-at-aurion.nl} 13, 21 -- Cc: Microscopy-at-microscopy.com 13, 21 -- From: Jan Leunissen {leunissen-at-aurion.nl} 13, 21 -- Subject: Re: [Microscopy] SEM Immunogold labeling & Albumin 13, 21 -- Date: Fri, 29 Sep 2006 09:16:08 +1200 13, 21 -- To: Mariangela.Fedel-at-apss.tn.it 13, 21 -- X-Mailer: Apple Mail (2.752.2) 13, 21 -- Content-Transfer-Encoding: 8bit 13, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8SLGEJW019859 ==============================End of - Headers==============================
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Title-Subject: [Filtered] Leica 1268 Ultrostainer spare parts
Question: I am in need of spare parts for the Leica 1268 Ultrostainer, the pump specifically, or even a used machine. Does anyone know of a resource I can contact? I would appreciate any leads.
Kathy Troughton University of Tennessee, Memphis ktrou-at-nb.utmem.edu 901-448-5976
A colleague wants to look at the microfibrillar structure of mature cotton fibres in the TEM and has asked us to look at the possibilities. While a literature search is being done, I was wondering if anyone in the microscopy community had done this previously. If so, what sample preparation techniques were used, what, if any staining was done and what are some of the pitfalls.....
Any help would be greatly 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
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============================== 10, 28 -- From Colin.Veitch-at-csiro.au Fri Sep 29 00:16:10 2006 10, 28 -- Received: from act-MTAout2.csiro.au (act-MTAout2.csiro.au [150.229.7.38]) 10, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8T5G9wZ015764 10, 28 -- for {microscopy-at-msa.microscopy.com} ; Fri, 29 Sep 2006 00:16:10 -0500 10, 28 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=t5kJTc0jpD2KFf/ftT6z3okwl0V2PbC8Ayie749J2Bp0osW8yZLg+US0GfX9PJ/lEPr9elqdLeq39ip4qKy4HSzMUa4zEqawIziyh9I3EpOUArwXJjN110nsH0tQ2sWT; 10, 28 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 10, 28 -- by act-MTAout2.csiro.au with ESMTP; 29 Sep 2006 15:15:28 +1000 10, 28 -- X-IronPort-AV: i="4.09,233,1157292000"; 10, 28 -- d="scan'208"; a="112889986:sNHT23080852" 10, 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); 10, 28 -- Fri, 29 Sep 2006 15:16:00 +1000 10, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 10, 28 -- content-class: urn:content-classes:message 10, 28 -- MIME-Version: 1.0 10, 28 -- Content-Type: text/plain; 10, 28 -- charset="us-ascii" 10, 28 -- Subject: Thin sectioning of cotton fibres 10, 28 -- Date: Fri, 29 Sep 2006 15:16:00 +1000 10, 28 -- Message-ID: {32CDDDAA7161394599F00254949157490248DF-at-exvic5-gex.nexus.csiro.au} 10, 28 -- X-MS-Has-Attach: 10, 28 -- X-MS-TNEF-Correlator: 10, 28 -- Thread-Topic: Thin sectioning of cotton fibres 10, 28 -- Thread-Index: AcbjhlgmbTjq9E8sQ0WdvH4K+K0Ecg== 10, 28 -- From: {Colin.Veitch-at-csiro.au} 10, 28 -- To: {microscopy-at-msa.microscopy.com} 10, 28 -- X-OriginalArrivalTime: 29 Sep 2006 05:16:00.0760 (UTC) FILETIME=[5A59DF80:01C6E386] 10, 28 -- Content-Transfer-Encoding: 8bit 10, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k8T5G9wZ015764 ==============================End of - Headers==============================
The Molecular Imaging Unit (MIU) and Cellular Imaging Unit (CIU) are organizing an international scientific symposium Advanced Technologies in Biological Imaging (October 26-27, 2006) in Biomedicum Helsinki, Finland. There is no registration fee or advance registration needed for the symposium.
For more information, visit MIU web page: http://www.miu.helsinki.fi/seminars.htm
Welcome to Helsinki!
-- Ville-Veikko Oksa, M.Sc. | Molecular Imaging Unit Imaging Analyst | Biomedicum Helsinki, Room B505b ville-veikko.oksa-at-helsinki.fi | PO Box 63 (Haartmaninkatu 8) Tel: +358-9-191-25579 (work) | FI-00014 University of Helsinki +358-40-744-3232 (mobile) | http://www.miu.helsinki.fi/
==============================Original Headers============================== 8, 28 -- From vvoksa-at-mappi.helsinki.fi Fri Sep 29 01:48:49 2006 8, 28 -- Received: from sender-02.it.helsinki.fi (sender-02.it.helsinki.fi [128.214.205.137]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8T6ml5P027506 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 29 Sep 2006 01:48:49 -0500 8, 28 -- Received: from quas.it.helsinki.fi (quas.it.helsinki.fi [128.214.205.29]) 8, 28 -- by sender-02.it.helsinki.fi (8.13.7/8.13.7) with ESMTP id k8T6mi2q011848 8, 28 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT); 8, 28 -- Fri, 29 Sep 2006 09:48:44 +0300 8, 28 -- Received: from quas.it.helsinki.fi (localhost.localdomain [127.0.0.1]) 8, 28 -- by quas.it.helsinki.fi (8.13.1/8.13.1) with ESMTP id k8T6lRqC015323; 8, 28 -- Fri, 29 Sep 2006 09:47:27 +0300 8, 28 -- Received: (from apache-at-localhost) 8, 28 -- by quas.it.helsinki.fi (8.13.1/8.13.1/Submit) id k8T6lRsF015319; 8, 28 -- Fri, 29 Sep 2006 09:47:27 +0300 8, 28 -- Received: from 128.214.167.37 ( [128.214.167.37]) 8, 28 -- as user vvoksa-at-localhost by www1.helsinki.fi with HTTP; 8, 28 -- Fri, 29 Sep 2006 09:47:27 +0300 8, 28 -- Message-ID: {1159512447.451cc17f6dc67-at-www1.helsinki.fi} 8, 28 -- Date: Fri, 29 Sep 2006 09:47:27 +0300 8, 28 -- From: Ville-Veikko Oksa {ville-veikko.oksa-at-helsinki.fi} 8, 28 -- To: Microscopy-at-microscopy.com, CONFOCAL-at-LISTSERV.BUFFALO.EDU 8, 28 -- Subject: Scientific Symposium: Advanced Technologies in Biological Imaging, October 8, 28 -- MIME-Version: 1.0 8, 28 -- Content-Type: text/plain; charset=us-ascii 8, 28 -- Content-Transfer-Encoding: 7bit 8, 28 -- User-Agent: Internet Messaging Program (IMP) 3.1 8, 28 -- X-Originating-IP: 128.214.167.37 8, 28 -- Sender: vvoksa-at-mappi.helsinki.fi ==============================End of - Headers==============================
Hello, We have a following trouble with LKB knifemaker 7801A. We cannot remove clamping head from the knifemaker body, even if we exactly follow the procedure in operation manual. Something is blocking the clamping head on the bracket. Did anybody have similar problem and found the resolution? Thanking you in advance for any suggestion and advices. Best regards from Prague Oldrich
-- Oldrich Benada Institute of Microbiology Acad. Sci. CR Videnska 1083 CZ-142 00 Prague 4 Czech Republic
==============================Original Headers============================== 3, 22 -- From benada-at-biomed.cas.cz Fri Sep 29 03:54:05 2006 3, 22 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 3, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8T8s48B007573 3, 22 -- for {microscopy-at-microscopy.com} ; Fri, 29 Sep 2006 03:54:05 -0500 3, 22 -- Received: from mail2.biomed.cas.cz (localhost [127.0.0.1]) 3, 22 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id k8T8o4dI021459 3, 22 -- for {microscopy-at-microscopy.com} ; Fri, 29 Sep 2006 10:50:05 +0200 (CEST) 3, 22 -- Received: from 147.231.44.104 3, 22 -- (SquirrelMail authenticated user benada) 3, 22 -- by mail2.biomed.cas.cz with HTTP; 3, 22 -- Fri, 29 Sep 2006 10:50:05 +0200 (CEST) 3, 22 -- Message-ID: {1512.147.231.44.104.1159519805.squirrel-at-mail2.biomed.cas.cz} 3, 22 -- Date: Fri, 29 Sep 2006 10:50:05 +0200 (CEST) 3, 22 -- Subject: LKB knifemaker 7801A problem 3, 22 -- From: "Oldrich Benada" {benada-at-biomed.cas.cz} 3, 22 -- To: microscopy-at-microscopy.com 3, 22 -- User-Agent: SquirrelMail/1.4.6 3, 22 -- MIME-Version: 1.0 3, 22 -- Content-Type: text/plain;charset=iso-8859-2 3, 22 -- Content-Transfer-Encoding: 8bit 3, 22 -- X-Priority: 3 (Normal) 3, 22 -- Importance: Normal ==============================End of - Headers==============================
The Analytical Imaging Facility, a comprehensive microscopy core facility at the Albert Einstein College of Medicine in New York, has an opening for a Biological EM Technician.
Please see the MSA Placement Office/Job Postings for details of the job description. http://www.microscopy.org/
**************************************************************************** Frank Macaluso, Director phone: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
==============================Original Headers============================== 6, 28 -- From macaluso-at-aecom.yu.edu Fri Sep 29 12:44:09 2006 6, 28 -- Received: from mailgw.aecom.yu.edu (mailgw.aecom.yu.edu [129.98.1.16]) 6, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8THi87d027206 6, 28 -- for {microscopy-at-microscopy.com} ; Fri, 29 Sep 2006 12:44:08 -0500 6, 28 -- Received: from draco.aecom.yu.edu (draco.aecom.yu.edu [129.98.1.160]) 6, 28 -- by mailgw.aecom.yu.edu (8.12.11.20060308/8.12.11) with ESMTP id k8THi8Ox011627 6, 28 -- for {microscopy-at-microscopy.com} ; Fri, 29 Sep 2006 13:44:08 -0400 6, 28 -- Received: from draco.aecom.yu.edu (unknown [127.0.0.1]) 6, 28 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id DE123329962 6, 28 -- for {microscopy-at-microscopy.com} ; Fri, 29 Sep 2006 13:41:02 -0400 (EDT) 6, 28 -- X-AuditID: 816201a0-ad769bb000004903-d9-451d5aaeac38 6, 28 -- Received: from post.aecom.yu.edu (post.aecom.yu.edu [129.98.1.100]) 6, 28 -- by draco.aecom.yu.edu (Symantec Mail Security) with ESMTP id A38242E5726 6, 28 -- for {microscopy-at-microscopy.com} ; Fri, 29 Sep 2006 13:41:02 -0400 (EDT) 6, 28 -- Received: from aif1.aecom.yu.edu (aif1.aif.aecom.yu.edu [129.98.80.55]) 6, 28 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 6, 28 -- (No client certificate requested) 6, 28 -- by post.aecom.yu.edu (Postfix) with ESMTP id 1568329; 6, 28 -- Fri, 29 Sep 2006 13:44:08 -0400 (EDT) 6, 28 -- Message-Id: {7.0.1.0.0.20060929132149.01b51a60-at-aecom.yu.edu} 6, 28 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 6, 28 -- Date: Fri, 29 Sep 2006 13:42:06 -0400 6, 28 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 6, 28 -- From: Frank Macaluso {macaluso-at-aecom.yu.edu} 6, 28 -- Subject: Job Opening EM Technician 6, 28 -- Mime-Version: 1.0 6, 28 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 6, 28 -- X-Brightmail-Tracker: AAAAAA== ==============================End of - Headers==============================
Colin, The essence of the problem is that heavy metals do not stain polysaccharides reliably. Cell walls in TEM often look textured and these textures are often ascribed to microfibrils, but given that UA and Lead don't stain cellulose some authors have argued that the textures arise from precipitation artifacts, not actual cell wall texture. I find these arguments persuasive and feel that one must be cautious and circumspect in interpreting typically ultrathin sections wrt microfibril structure and orientation.
Some of the ways to enhance contrast involve various pre embedding or post embedding extractions, some of which definitely change microfibril structure. Some authors have etched the resin and then shadowed. The shadowing allows microfibrils to show up directly but etching spurs or epon is pretty tough treatment and could well expand and muck up the microfibrils. Just for fun, I'll suggest that my good old friend butyl methyl methacrylate might be interesting here, because it is etchable with only a 10 min acetone treatment, and hence unlikely to derange the microfibrils. That could then be shadowed and examined. I have no idea at all if that would work so go ahead and laugh. Its OK.
If you have access to an FESEM or an AFM, you might think about cutting the fibers in the fresh state obliquely and removing the cytoplasm and then prepping them for fesem or afm and looking at the innermost surface. That would only allow a small region to be examined but I think with reasonable confidence about what one was seeing.
If the only issue is fiber angle, then a lot can be done with good old xrays.
Hope this helps.
Tobias
} } Hi, } } A colleague wants to look at the microfibrillar structure of mature } cotton fibres in the TEM and has asked us to look at the possibilities. } While a literature search is being done, I was wondering if anyone in } the microscopy community had done this previously. If so, what sample } preparation techniques were used, what, if any staining was done and } what are some of the pitfalls..... } } Any help would be greatly 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. }
Colorado School of Mines Metallurgical and Materials Engineering Professor, Electron Microscopy
The Colorado School of Mines invites applications for an anticipated Professor position in the Metallurgical and Materials Engineering Department in the area of materials characterization. The successful candidate will serve as the Director of the Electron Microscopy Laboratory. A Ph.D. in materials science, metallurgy, ceramics or closely related field is required. Appointment at the Associate Professor level may be considered.
For a complete job announcement, more information about the position and the university, and instructions on how to apply, please visit our web site at:
http://csmis15.is.mines.edu/hr/Faculty_Jobs.shtm
CSM is an EEO/AA employer and is committed to enhancing the diversity of its campus community. Women, minorities, veterans, and individuals with disabilities are encouraged to apply.
Please send all inquiries to the address listed in the online announcement.
John Chandler johnpchandler-at-comcast.net
==============================Original Headers============================== 9, 16 -- From johnpchandler-at-comcast.net Fri Sep 29 14:33:49 2006 9, 16 -- Received: from rwcrmhc15.comcast.net (rwcrmhc15.comcast.net [216.148.227.155]) 9, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8TJXmda018201 9, 16 -- for {Microscopy-at-microscopy.com} ; Fri, 29 Sep 2006 14:33:49 -0500 9, 16 -- Received: from rmailcenter03.comcast.net ([204.127.197.113]) 9, 16 -- by comcast.net (rwcrmhc15) with SMTP 9, 16 -- id {20060929193347m1500lh4q4e} ; Fri, 29 Sep 2006 19:33:47 +0000 9, 16 -- Received: from [138.67.28.173] by rmailcenter03.comcast.net; 9, 16 -- Fri, 29 Sep 2006 19:33:47 +0000 9, 16 -- From: johnpchandler-at-comcast.net 9, 16 -- To: Microscopy-at-microscopy.com (MicroscopyList) 9, 16 -- Subject: Faculty position in EM at Colorado School of Mines 9, 16 -- Date: Fri, 29 Sep 2006 19:33:47 +0000 9, 16 -- Message-Id: {092920061933.24463.451D751B000008CD00005F8F22007456729D0A040B020E080C9F02080106-at-comcast.net} 9, 16 -- X-Mailer: AT&T Message Center Version 1 (Apr 11 2006) 9, 16 -- X-Authenticated-Sender: am9obnBjaGFuZGxlckBjb21jYXN0Lm5ldA== ==============================End of - Headers==============================
Good question, and it does seem unlikely that many High Schools have their own Electron Microsocpes. However, in the Dayton Ohio area, Wright Patterson Air Force Base (WPAFB) Educational Outreach Office does run a program called TECH Trek Mobile Research laboratory. It is basically a mobile home with an SEM and other high tech microscopes complete with all the accessories (including a generator) that they bring to K- 12 schools and leave (for a week I think). They train the teachers to run the scopes and leave them to it!
http://edoutreach.wpafb.af.mil/
On 28 Sep 2006 at 9:15, 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 } ---------------------------------------------------------------------------- } } I've been asked several times the following question: } } How many high schools in the world actually have equipment such as SEM's } or other fairly sophisticated scientific equipment for use in the } classroom? } } If you are a high school, or know of a high school that has this kind of } capability, can you please post that information. I would like to know } where you are, contact information, and what equipment you use. } } I think it will be of interest for our whole scientific community to know } the answer to that question, so I suggest that you post the answer to the } group as a whole. } } If you prefer to answer me directly, please indicate if you prefer } information to be shared or not as the case may be. } } Thank you, } } dj } } } } } ==============================Original Headers============================== } 10, 16 -- From dljones-at-bestweb.net Thu Sep 28 09:15:24 2006 } 10, 16 -- Received: from vms044pub.verizon.net (vms044pub.verizon.net [206.46.252.44]) } 10, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8SEFONA006752 } 10, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 28 Sep 2006 09:15:24 -0500 } 10, 16 -- Received: from localhost ([71.247.114.109]) } 10, 16 -- by vms044.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep } 10, 16 -- 9 2005)) with ESMTPA id {0J6B00G7F3LEFSQ0-at-vms044.mailsrvcs.net} for } 10, 16 -- Microscopy-at-microscopy.com; Thu, 28 Sep 2006 09:15:18 -0500 (CDT) } 10, 16 -- Date: Thu, 28 Sep 2006 10:27:44 -0400 (Eastern Standard Time) } 10, 16 -- From: "David L. Jones" {dljones-at-bestweb.net} } 10, 16 -- Subject: Perspectives: High School Microscopy } 10, 16 -- X-X-Sender: dljones-at-pop-croton.bestweb.net } 10, 16 -- To: Microscopy-at-microscopy.com } 10, 16 -- Message-id: {Pine.WNT.4.64.0609281008410.3292-at-H-F1} } 10, 16 -- MIME-version: 1.0 } 10, 16 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed } ==============================End of - Headers==============================
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 9, 27 -- From edelmare-at-muohio.edu Fri Sep 29 15:42:28 2006 9, 27 -- Received: from mulnx11.mcs.muohio.edu (mulnx11.mcs.muohio.edu [134.53.6.66]) 9, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8TKgSpZ029837 9, 27 -- for {microscopy-at-Microscopy.com} ; Fri, 29 Sep 2006 15:42:28 -0500 9, 27 -- Received: from mulnx23.mcs.muohio.edu (mulnx23.mcs.muohio.edu [134.53.6.10]) 9, 27 -- by mulnx11.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k8TKgPFp009780; 9, 27 -- Fri, 29 Sep 2006 16:42:25 -0400 9, 27 -- Received: from [192.168.1.23] ([134.53.14.97]) 9, 27 -- by mulnx23.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k8TKgPoW016718; 9, 27 -- Fri, 29 Sep 2006 16:42:25 -0400 9, 27 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 9, 27 -- To: dljones-at-bestweb.net 9, 27 -- Date: Fri, 29 Sep 2006 16:42:26 -0400 9, 27 -- MIME-Version: 1.0 9, 27 -- Subject: Re: [Microscopy] Perspectives: High School Microscopy 9, 27 -- CC: microscopy-at-Microscopy.com 9, 27 -- Message-ID: {451D4CF2.29330.23EF6EE7-at-edelmare.muohio.edu} 9, 27 -- Priority: normal 9, 27 -- In-reply-to: {200609281415.k8SEFxri008054-at-ns.microscopy.com} 9, 27 -- References: {200609281415.k8SEFxri008054-at-ns.microscopy.com} 9, 27 -- X-mailer: Pegasus Mail for Windows (4.41) 9, 27 -- Content-type: text/plain; charset=US-ASCII 9, 27 -- Content-transfer-encoding: 7BIT 9, 27 -- Content-description: Mail message body 9, 27 -- X-Real-ConnectIP: 134.53.14.97 9, 27 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.66 9, 27 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.10 ==============================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 klm-at-berkeley.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: klm-at-berkeley.edu Name: Kent McDonald
Organization: University of California Berkeley
Title-Subject: [Filtered] Biological Cryo EM Course at UC Berkeley
Question: The Elecron Microscope Laboratory at the University of California, Berkeley, will be holding a Cryo EM Course from January 7-12, 2007. The course is for biological research scientists who already have EM experience but who want to learn more about advanced cryotechniques for EM of cells and tissues. We will focus on high pressure freezing (all three commercial machines will be available), vitreous cryosectioning, Tokuyasu cryosectioning and immunolabeling, and an Introduction to Cellular Tomography.
For more information including the Course Schedule and Registration forms, go to: http://em-lab.berkeley.edu/EML/class/cryoem-course.php
Registration will be limited to 12 because of the hands-on nature of the course. There are currently 4 openings remaining.
We plan to buy a cryo-ultramicrotome for polymer TEM sample preparation. Is anybody familiar with RMC MT-X cryo-ultramicrotome? Any suggestion or any comments for other cryo-ultramicrotome would be greatly appreciated.
Thanks,
Zhiqiang Chen Institute of Advance Materials and New Energy University of Louiville Louisville, KY 40292
==============================Original Headers============================== 4, 26 -- From z0chen09-at-louisville.edu Sun Oct 1 08:33:07 2006 4, 26 -- Received: from erouter2.it-datacntr.louisville.edu (erouter2.it-datacntr.louisville.edu [136.165.237.35]) 4, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k91DX794006592 4, 26 -- for {Microscopy-at-microscopy.com} ; Sun, 1 Oct 2006 08:33:07 -0500 4, 26 -- Received: from PostX2 (postx2.it-servers.louisville.edu [136.165.233.129]) 4, 26 -- by erouter2.it-datacntr.louisville.edu (Postfix) with ESMTP id 8EE957244D 4, 26 -- for {Microscopy-at-microscopy.com} ; Sun, 1 Oct 2006 09:30:25 -0400 (EDT) 4, 26 -- Received: from savgw-out.louisville.edu ([136.165.234.42]) 4, 26 -- by PostX2 (PostX Enterprise 6.0.1 SMTP Adapter) with SMTP ID 421 4, 26 -- for {Microscopy-at-microscopy.com} ; 4, 26 -- Sun, 1 Oct 2006 09:18:18 -0400 (EDT) 4, 26 -- Received: from gwise.louisville.edu ([136.165.232.131]) 4, 26 -- by savgw-out.louisville.edu (SAVSMTP 3.1.7.47) with SMTP id M2006100109294129692 4, 26 -- for {Microscopy-at-microscopy.com} ; Sun, 01 Oct 2006 09:29:41 -0400 4, 26 -- Received: from sgate-MTA by gwise.louisville.edu 4, 26 -- with Novell_GroupWise; Sun, 01 Oct 2006 09:30:25 -0400 4, 26 -- Message-Id: {451F8AA90200005C0000270E-at-gwise.louisville.edu} 4, 26 -- X-Mailer: Novell GroupWise Internet Agent 7.0.1 4, 26 -- Date: Sun, 01 Oct 2006 09:30:17 -0400 4, 26 -- From: "Zhiqiang Chen" {z0chen09-at-louisville.edu} 4, 26 -- To: {Microscopy-at-microscopy.com} 4, 26 -- Subject: cryo-ultramicrotome 4, 26 -- Mime-Version: 1.0 4, 26 -- Content-Type: text/plain; charset=US-ASCII 4, 26 -- Content-Transfer-Encoding: 7bit 4, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
X-from: Ken Converse [mailto:kenconverse-at-qualityimages.biz] Sent: Monday, October 02, 2006 11:47 AM To: 'plarson-at-ou.edu'
Hi everyone,
I was wondering if anyone on the board had any advice for venting electron microscope columns/sample chambers. In particular, we want a system that allows the column or sample chamber to reach atmospheric pressure by venting with dry N2 but does not allow the chamber to over pressurize above atmosphere to protect seals, thin window on EDS detectors, etc.
I'm aware that there are on-demand gas regulators that can control this sort of thing. Does anyone have any suggestions for a good vendor or model number for this sort of regulator?
Alternatively, I've also heard that demand valves that divers use are good for this sort of application as it only supplies gas when the diver
breathes or sucks through the mouthpiece (or in this case when the chamber is still under vacuum). Does anyone know a particular brand or type of diver demand valve that works well for this application and is easy to modify (i.e. relatively easy to install standard fittings on the
ends like NPT, etc.)?
Any comments or suggestions would be greatly appreciated. Thanks in advance,
preston
-- Preston Larson Research Scientist Samuel Roberts Noble Electron Microscopy Laboratory 770 Van Vleet Oval Phone: (405) 325-4391
==============================Original Headers============================== 9, 17 -- From plarson-at-ou.edu Wed Sep 27 10:29:15 2006 9, 17 -- Received: from c3p0.ou.edu (mail.zero.ou.edu [129.15.0.75]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8RFTFwv029604 9, 17 -- for {Microscopy-at-Microscopy.Com} ; Wed, 27 Sep 2006 10:29:15 -0500 9, 17 -- Received: from [127.0.0.1] ([129.15.39.56]) 9, 17 -- by c3p0.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 9, 17 -- 2005)) with ESMTP id {0J690035JCCPEI80-at-c3p0.ou.edu} for 9, 17 -- Microscopy-at-Microscopy.Com; Wed, 27 Sep 2006 10:29:13 -0500 (CDT) 9, 17 -- Date: Wed, 27 Sep 2006 10:29:11 -0500 9, 17 -- From: Preston Larson {plarson-at-ou.edu} 9, 17 --
DJ, In November 2000 I donated an ETEC Autoscan to the Red Lion Area Sr. High School in Red Lion, PA along with a very old (vacuum tube logic) Varian vacuum evaporator. They have been using it ever since and I have been providing free service.
This was the school system that my kids went to and the deal was that I would treat them as a full service contract for at least as long as my kids were in the school system. They're all graduated and I've since moved to Maine, but continue to service the scope, just not promising as prompt service due to the current distance involved.
IMHO the problem isn't getting the equipment. It's really not all that unusual for this stuff to come up free. The problem is service. I would encourage anyone who is familiar with servicing SEMs, TEMs and other "high tech" equipment to adopt a school, find them some free equipment and give them free service. If you want to get kids interested in the sciences then they need to have the opportunity to play with the toys that scientists play with. After all, many of us were drawn to our areas of interest by the cool toys.
Ken Converse owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: dljones-at-bestweb.net [mailto:dljones-at-bestweb.net] Sent: Thursday, September 28, 2006 10:18 AM To: kenconverse-at-qualityimages.biz
I've been asked several times the following question:
How many high schools in the world actually have equipment such as SEM's
or other fairly sophisticated scientific equipment for use in the classroom?
If you are a high school, or know of a high school that has this kind of
capability, can you please post that information. I would like to know where you are, contact information, and what equipment you use.
I think it will be of interest for our whole scientific community to know the answer to that question, so I suggest that you post the answer to the group as a whole.
If you prefer to answer me directly, please indicate if you prefer information to be shared or not as the case may be.
Thank you,
dj
==============================Original Headers============================== 10, 16 -- From dljones-at-bestweb.net Thu Sep 28 09:15:24 2006 10, 16 -- Received: from vms044pub.verizon.net (vms044pub.verizon.net [206.46.252.44]) 10, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k8SEFONA006752 10, 16 -- for {Microscopy-at-microscopy.com} ; Thu, 28 Sep 2006 09:15:24 -0500 10, 16 -- Received: from localhost ([71.247.114.109]) 10, 16 -- by vms044.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep 10, 16 -- 9 2005)) with ESMTPA id {0J6B00G7F3LEFSQ0-at-vms044.mailsrvcs.net} for 10, 16 -- Microscopy-at-microscopy.com; Thu, 28 Sep 2006 09:15:18 -0500 (CDT) 10, 16 -- Date: Thu, 28 Sep 2006 10:27:44 -0400 (Eastern Standard Time) 10, 16 -- From: "David L. Jones" {dljones-at-bestweb.net} 10, 16 -- Subject: Perspectives: High School Microscopy 10, 16 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 10, 16 -- To: Microscopy-at-microscopy.com 10, 16 -- Message-id: {Pine.WNT.4.64.0609281008410.3292-at-H-F1} 10, 16 -- MIME-version: 1.0 10, 16 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed ==============================End of - Headers==============================
==============================Original Headers============================== 26, 23 -- From kenconverse-at-qualityimages.biz Mon Oct 2 11:42:16 2006 26, 23 -- Received: from mta9.adelphia.net (mta9.adelphia.net [68.168.78.199]) 26, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k92GgG6l023112 26, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 2 Oct 2006 11:42:16 -0500 26, 23 -- Received: from Ken ([24.53.5.33]) by mta9.adelphia.net 26, 23 -- (InterMail vM.6.01.05.02 201-2131-123-102-20050715) with ESMTP 26, 23 -- id {20061002164216.XZTT8494.mta9.adelphia.net-at-Ken} ; 26, 23 -- Mon, 2 Oct 2006 12:42:16 -0400 26, 23 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 26, 23 -- To: {dljones-at-bestweb.net} , "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 26, 23 -- Subject: RE: [Microscopy] Perspectives: High School Microscopy 26, 23 -- Date: Mon, 2 Oct 2006 12:42:03 -0400 26, 23 -- Message-ID: {003801c6e641$b0b59230$6401a8c0-at-Ken} 26, 23 -- MIME-Version: 1.0 26, 23 -- Content-Type: text/plain; 26, 23 -- charset="us-ascii" 26, 23 -- Content-Transfer-Encoding: 7bit 26, 23 -- X-Priority: 3 (Normal) 26, 23 -- X-MSMail-Priority: Normal 26, 23 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 26, 23 -- In-Reply-To: {200609281417.k8SEHkNk011616-at-ns.microscopy.com} 26, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 26, 23 -- Importance: Normal ==============================End of - Headers==============================
a: find an exhaustive listing of the Smithsonian reference materials (and their reference compositions)
b: find out exactly how I can lay my hands, no, my tweezers, on some of them?
I have searched the Smithsonian website but to no avail.
all the best
Ritchie
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 10, 26 -- From r.sims-at-auckland.ac.nz Mon Oct 2 16:30:31 2006 10, 26 -- Received: from groucho.itss.auckland.ac.nz (mailhost.auckland.ac.nz [130.216.190.11]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k92LUUJN006602 10, 26 -- for {Microscopy-at-microscopy.com} ; Mon, 2 Oct 2006 16:30:31 -0500 10, 26 -- Received: from localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) 10, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 6076F352AB 10, 26 -- for {Microscopy-at-microscopy.com} ; Tue, 3 Oct 2006 10:30:29 +1300 (NZDT) 10, 26 -- Received: from groucho.itss.auckland.ac.nz ([127.0.0.1]) 10, 26 -- by localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 10, 26 -- with ESMTP id 13659-22 for {Microscopy-at-microscopy.com} ; 10, 26 -- Tue, 3 Oct 2006 10:30:29 +1300 (NZDT) 10, 26 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 10, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 325CC350CA 10, 26 -- for {Microscopy-at-microscopy.com} ; Tue, 3 Oct 2006 10:30:28 +1300 (NZDT) 10, 26 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 10, 26 -- To: Microscopy-at-microscopy.com 10, 26 -- Date: Tue, 03 Oct 2006 10:35:08 +1300 10, 26 -- MIME-Version: 1.0 10, 26 -- Subject: Smithsonian Microbeam Reference Materials 10, 26 -- Message-ID: {45223CDC.30160.A3CF8C-at-localhost} 10, 26 -- Priority: normal 10, 26 -- X-mailer: Pegasus Mail for Windows (4.21c) 10, 26 -- Content-type: text/plain; charset=US-ASCII 10, 26 -- Content-transfer-encoding: 7BIT 10, 26 -- Content-description: Mail message body 10, 26 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
Gene Jarosowich wrote a short article on the history of the Smithsonian standards in the NIST Journal of Research (proceedings of a NIST-MAS topical workshop that is well worth looking at, about accuracy in epma) in 2002 -- that has an appendix with a list of the standards. You can download that article and others from my probe class web site www.geology.wisc.edu/~johnf/g777/777NISTarticles.html the fifth article down.
John
-- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
"The first rule of all intelligent tinkering is to save every cog and wheel." -- Aldo Leopold
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." -- Richard P. Feynman
==============================Original Headers============================== 7, 25 -- From johnf-at-geology.wisc.edu Mon Oct 2 16:57:51 2006 7, 25 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k92Lvobj017480 7, 25 -- for {microscopy-at-microscopy.com} ; Mon, 2 Oct 2006 16:57:50 -0500 7, 25 -- Received: from localhost (localhost [127.0.0.1]) 7, 25 -- by localhost (Postfix) with ESMTP id 7A99B20D08; 7, 25 -- Mon, 2 Oct 2006 16:57:50 -0500 (CDT) 7, 25 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 7, 25 -- by localhost (geology.wisc.edu [127.0.0.1]) (amavisd-new, port 10024) 7, 25 -- with ESMTP id 05770-09; Mon, 2 Oct 2006 16:57:47 -0500 (CDT) 7, 25 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 7, 25 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 7, 25 -- (No client certificate requested) 7, 25 -- by ice.geology.wisc.edu (Postfix) with ESMTP id 149A320D1C; 7, 25 -- Mon, 2 Oct 2006 16:57:47 -0500 (CDT) 7, 25 -- Mime-Version: 1.0 7, 25 -- Message-Id: {p0623090ac1473a74bfd4-at-[144.92.206.57]} 7, 25 -- In-Reply-To: {200610022136.k92LaZWI014959-at-ns.microscopy.com} 7, 25 -- References: {200610022136.k92LaZWI014959-at-ns.microscopy.com} 7, 25 -- Date: Mon, 2 Oct 2006 16:56:42 -0500 7, 25 -- To: r.sims-at-auckland.ac.nz, microscopy-at-microscopy.com 7, 25 -- From: John Fournelle {johnf-at-geology.wisc.edu} 7, 25 -- Subject: Re: [Microscopy] Smithsonian Microbeam Reference Materials 7, 25 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 25 -- X-Virus-Scanned: amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
To Owners/Users of Sorvall MT2B or RMC MT2C ultramicrotomes:
Does anyone have any current information regarding third-party repair services in the St. Louis/Kansas City/Chicago area? We need someone that will do the service onsite as we have three or four machines requiring maintenance.
Thank you so much,
Jaclynn Lett Senior Research Technician Research Center for Auditory and Vestibular Studies Department of Otolaryngology Washington University School of Medicine 660 S. Euclid Ave., Campus Box 8115 St. Louis, MO 63110
Voice: 314-747-7257 Fax: 314-747-7230 Email: lettj-at-ent.wustl.edu http://www.otocore.org/ {br/} The materials in this message are private and may contain Protected Healthcare Information. If you are not the intended recipient, be advised that any unauthorized use, disclosure, copying or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this email in error, please immediately notify the sender via telephone or return mail.
==============================Original Headers============================== 7, 23 -- From LettJ-at-wusm-pcf.wustl.edu Tue Oct 3 11:43:16 2006 7, 23 -- Received: from MAIL4.wusm-pcf.wustl.edu (ip-17-171.wustl.edu [128.252.17.171]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k93GhGPY020654 7, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 3 Oct 2006 11:43:16 -0500 7, 23 -- Received: from EX02.wusm-pcf.wustl.edu ([10.39.162.137]) by MAIL4.wusm-pcf.wustl.edu with Microsoft SMTPSVC(6.0.3790.1830); 7, 23 -- Tue, 3 Oct 2006 11:43:16 -0500 7, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 23 -- Content-class: urn:content-classes:message 7, 23 -- MIME-Version: 1.0 7, 23 -- Content-Type: text/plain; 7, 23 -- charset="us-ascii" 7, 23 -- Subject: EQUIP: ultramicrotome repair 7, 23 -- Date: Tue, 3 Oct 2006 11:43:15 -0500 7, 23 -- Message-ID: {8AADB33205E9CE49BB22275C309FABDA0B5428-at-EX02.wusm-pcf.wustl.edu} 7, 23 -- X-MS-Has-Attach: 7, 23 -- X-MS-TNEF-Correlator: 7, 23 -- Thread-Topic: EQUIP: ultramicrotome repair 7, 23 -- Thread-Index: AcbnCwWoAXNN06/PQuWGMhqvIi2ygQ== 7, 23 -- From: "Lett, Jaclynn" {LettJ-at-ent.wustl.edu} 7, 23 -- To: {Microscopy-at-MSA.Microscopy.Com} 7, 23 -- X-OriginalArrivalTime: 03 Oct 2006 16:43:16.0273 (UTC) FILETIME=[0648D210:01C6E70B] 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k93GhGPY020654 ==============================End of - Headers==============================
Colin, I suspect you can get strong thin-section staining of cellulose and other polysaccharides with our favorite, KMnO4 followed by Pb (citrate ok, but we prefer Sato's Lead formula; Essential to avoid NMA (MNA?) in your epoxy mixture; best to use Araldite; my favorite is Araldite 506). I know this Mn-Pb sequence strongly stains the spiral chitin rib in micron and submicron tracheoles of insect muscle, and also stains the dextran (MW 500,000; lovely little wormy threads excluded to the extra-fibrillar space) we've used at 3% to osmotically squeeze the myofilament lattice in the glycerol-detergent-permeabilized muscle fibers. During aqueous fixation, the dextran is lost, rinsed away, but is nicely preserved in situ by freeze-substitution. I think it may even stain, though with less contrast, using the lead stain alone, come to think of it. I seem to recall that lead citrate alone stains glycogen, something I never see any more when working with permeabilized muscle.
Part of our staining success (and certainly our ultimate high contrast) might also be due to our preferred 2-step fixation (works for both aqueous and in -80 acetone for freeze-substitution) using tannic acid followed by uranyl acetate. No additional fix is needed for extracellular structures or well-anchored internal components of permeabilized cells; but you can use OsO4 after TA instead of UrAc. However, OsO4 toxicity and vapor makes it less popular with us than the equally effective UrAc.
I tried Googling got 60 seconds on permanganate staining of cellulose and it looks promising; hit #7 includes "becomes visible as permanganate is a stain for cellulose fibrils ... ". this 1979 reference attributes a 1975 paper DESHPANDE, B. P. (1976). Observations on the fine structure of plant cell walls. I. Use of permanganate staining. Ann. Bot. 40, 433-437.
I leave you to sift through the rest.
-mike reedy-
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
I'm sorry I didn't reply earlier, but if your model of LKB 7801A works like mine then I'm sure this will solve it.
A1. I assume that you have the black handled locking lever to 45 deg forward from vertical and the score selector to || //.
If this doesn't work then it's probably because the clamping head needs loosening off: I hope you have the same diagrams as me so (if not - get back to me): B1. Undo the silver cover (domed flat blade screw head) screw - labelled 7 in Fig 1. B2. using the correct hexagonal allen wrench loosen the screw inside the hole. B3. Now you can loosen the barrel of the locking head by putting something in one of 4 holes around the cylinder at the base of the shaft for the black handled locking lever. Turn it one or two rotations clockwise and you should see it loosen a bit.
Now if you go back to A1 above it should be possible to remove the head easily because it is not clamped so tightly.
If this works remember to re-adjust the barrel tension (B3 above) so that the black handled locking lever is clamped at about 90 to 100 deg from vertical. Effectively do B3, B2 and B1 in reverse.
Please come back to me if my instructions are too vague or vary from your model.
I have copied this to the microscopy list just in case anyone else has this problem.
Malcolm
Malcolm Haswell e.m. unit School of Health, Natural and Social Sciences Fleming Building University of Sunderland Tyne & Wear SR1 3SD UK e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- X-from: benada-at-biomed.cas.cz
Hi, All: Please let me know if any body has a second TEM with CCD for sale. Thank you for your attention.
Zhiqiang Chen Institute for Advance Materials and Renewable Energy University of Louisville Louisville, KY 40292
==============================Original Headers============================== 2, 26 -- From z0chen09-at-louisville.edu Wed Oct 4 16:01:44 2006 2, 26 -- Received: from erouter2.it-datacntr.louisville.edu (erouter2.it-datacntr.louisville.edu [136.165.237.35]) 2, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k94L1iVY017989 2, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 4 Oct 2006 16:01:44 -0500 2, 26 -- Received: from PostX2 (postx2.it-servers.louisville.edu [136.165.233.129]) 2, 26 -- by erouter2.it-datacntr.louisville.edu (Postfix) with ESMTP id 3E8BA71057 2, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 4 Oct 2006 16:57:22 -0400 (EDT) 2, 26 -- Received: from savgw-out.louisville.edu ([136.165.234.42]) 2, 26 -- by PostX2 (PostX Enterprise 6.0.1 SMTP Adapter) with SMTP ID 840 2, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; 2, 26 -- Wed, 4 Oct 2006 16:45:06 -0400 (EDT) 2, 26 -- Received: from gwise.louisville.edu ([136.165.232.131]) 2, 26 -- by savgw-out.louisville.edu (SAVSMTP 3.1.7.47) with SMTP id M2006100416564801610 2, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 04 Oct 2006 16:56:48 -0400 2, 26 -- Received: from sgate-MTA by gwise.louisville.edu 2, 26 -- with Novell_GroupWise; Wed, 04 Oct 2006 16:57:22 -0400 2, 26 -- Message-Id: {4523E7F5.B28C.005C.0-at-gwise.louisville.edu} 2, 26 -- X-Mailer: Novell GroupWise Internet Agent 7.0.1 2, 26 -- Date: Wed, 04 Oct 2006 16:57:08 -0400 2, 26 -- From: "Zhiqiang Chen" {z0chen09-at-louisville.edu} 2, 26 -- To: {Microscopy-at-MSA.Microscopy.Com} 2, 26 -- Subject: looking for second-hand TEM 2, 26 -- Mime-Version: 1.0 2, 26 -- Content-Type: text/plain; charset=US-ASCII 2, 26 -- Content-Transfer-Encoding: 7bit 2, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
I would like to look at actin structure in fish skin epithelial cells. I usually use 2%GA-2%PFA fixative and spurr's resin, but I think actin structure is not clear enough. If anyone knows or experienced well preserved morphology, please give me any advice.
Thank you so much.
--------------------- Fumi KATOH post doctoral research fellow St. Francis Xavier University fkatoh-at-stfx.ca
==============================Original Headers============================== 5, 20 -- From fkatoh-at-stfx.ca Wed Oct 4 18:34:47 2006 5, 20 -- Received: from xmail1.stfx.ca (xmail1.stfx.ca [141.109.221.24]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k94NYjUB031545 5, 20 -- for {Microscopy-at-Microscopy.Com} ; Wed, 4 Oct 2006 18:34:46 -0500 5, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 20 -- Content-class: urn:content-classes:message 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; 5, 20 -- charset="iso-8859-1" 5, 20 -- Subject: TEM-fixation for actin 5, 20 -- Date: Wed, 4 Oct 2006 20:32:43 -0300 5, 20 -- Message-ID: {D1DA484CA5D7D5499AB6BBB4BCA429175CA4EC-at-demetrius.ad.stfx.ca} 5, 20 -- X-MS-Has-Attach: 5, 20 -- X-MS-TNEF-Correlator: 5, 20 -- Thread-Topic: TEM-fixation for actin 5, 20 -- Thread-Index: AcboDWPzpDtFkkVpQOiZMlr4+eES/g== 5, 20 -- From: "Fumi Katoh" {fkatoh-at-stfx.ca} 5, 20 -- To: {Microscopy-at-Microscopy.Com} 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k94NYjUB031545 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both marie.johnson1-at-att.net as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: marie.johnson1-at-att.net Name: Marie
Organization: Continental Carbon Company
Title-Subject: [Filtered] CamScan Model Cs44
Question: Does anyone have info on this SEM? Specs/year of manufacture? Thank you, Marie
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 burrmich-at-msu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: burrmich-at-msu.edu Name: Mike
Title-Subject: [Filtered] SEM (filament/image issues)
Question: Please help a newbie out a little...
Our regular SEM operator isn't here anymore (can't contact them) so i'm now in charge of running it. (used it sparingly in college)
We have a Hitachi S-570 (yeah a very old one). When i first started it up after about a month downtime, the filament was burnt. So i replaced the filament according to the settings in the manual. Well after starting it up again, i used the filament image mode to find the saturation point and everything. my settings were 20KV and WD of 30mm. I have a fairly circular filament image when the current gets up to only 60 micro amps... then there is a totally seperate half circle pertruding out from the side of the screen. When i turn the fillament knob to get a current above about 80, the image stays the same shape but starts to dissapear.
So when i go back to the best filament image (only 50 micro amps) and switch to normal mode to see an image, i can only see a very faint outline of my specimen. Focusing and gun allignments do nothing.
I'm pretty sure my problem has to do with when i set the new filament, but it was done exactly by the manual.
Electron/Ion Beam Instrument Engineer The University of Oregon's Center for Advanced Materials Characterization in Oregon (CAMCOR) is seeking applications for a full time staff position to begin January 2007. A strong background in maintaining, trouble shooting and upgrading electron/ion beam instruments and associated high voltage, vacuum, mechanical and electrical systems is required. Experience x-ray diffraction instrumentation is also desirable.
This position will be located in the new Lorry Lokey Integrated Science Laboratory, a state of the art nano and micro science analytical instrument facility designed specifically for exceptional nano-science performance. It will house the latest electron, ion and x-ray beam instrumentation available including a Zeiss Ultra TFEM, FEI Quanta 200 E-SEM, Cameca SX50 and SX100 microprobes, Ion-TOF SIMS, Philips Auger Spectrometer, Philips TEM, Kratos XPS and various assorted coaters, etchers, and other vacuum deposition systems.
The successful candidate with have a BS in a beam microscopy related field and an extensive background in instrument field service with significant practical experience troubleshooting high vacuum electron and ion beam instrumentation at both the system and PC board levels. Must be able to read and understand schematics for electronic circuits and systems. The successful applicant will be involved in modifying/improving instrumentation capabilities to enable the equipment to more fully support unique research needs and will be expected to work intimately with the scientific staff and research faculty. We seek candidates with a demonstrated commitment to working effectively with students, faculty and staff from diverse backgrounds.
Interested persons should send a resume with a detailed description of work experience and skills, and arrange for two letters of recommendation to be sent to: CAMCOR Instrument Engineer Search Committee, 1253 University of Oregon, Eugene, OR, 97403-1253. To be assured of full consideration, application materials must be received by November 1, 2006, but the search will remain open until the position is filled. For further information, contact John Donovan (donovan-at-uoregon.edu).
University of Oregon is an AA/EEO employer committed to cultural diversity.
==============================Original Headers============================== 6, 21 -- From donovan-at-uoregon.edu Thu Oct 5 12:31:47 2006 6, 21 -- Received: from smtp.uoregon.edu (mserv3.uoregon.edu [128.223.142.101]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95HVkqZ016427 6, 21 -- for {microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 12:31:47 -0500 6, 21 -- Received: from Probev.uoregon.edu (probev.uoregon.edu [128.223.92.75]) 6, 21 -- (authenticated bits=0) 6, 21 -- by smtp.uoregon.edu (8.13.7/8.13.7) with ESMTP id k95HVkVn032595 6, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 6, 21 -- for {microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 10:31:46 -0700 6, 21 -- Message-Id: {7.0.1.0.0.20061005103033.00f89840-at-uoregon.edu} 6, 21 -- Message-Id: {7.0.1.0.0.20061004112624.039222b0-at-uoregon.edu} 6, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 6, 21 -- Date: Thu, 05 Oct 2006 10:30:39 -0700 6, 21 -- To: microscopy-at-microscopy.com 6, 21 -- From: John Donovan {donovan-at-uoregon.edu} 6, 21 -- Subject: Full Time Instrument Engineer Position at the University of 6, 21 -- Oregon 6, 21 -- Mime-Version: 1.0 6, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 6, 21 -- X-Virus-Scanned: ClamAV 0.88.4/1998/Thu Oct 5 07:55:28 2006 on mserv3 6, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
I am happy to announce that DVDs of the 2006 MSA tutorials are now available. John Mansfield and his crew have done a great job once again. For full information about purchasing these DVDs or any others in the collection, please go to the MSA web site (www.microscopy.org) and look under REFERENCE for the video library.
Greg Erdos
==============================Original Headers============================== 3, 21 -- From gwe-at-ufl.edu Thu Oct 5 13:31:00 2006 3, 21 -- Received: from smtp.ufl.edu (smtp01.osg.ufl.edu [128.227.74.149]) 3, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95IUx4d027920 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 13:31:00 -0500 3, 21 -- Received: from [10.228.0.31] (ssrb-vpn1-0-31.vpn.ufl.edu [10.228.0.31]) 3, 21 -- (authenticated bits=0) 3, 21 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k95IUvCq3047440 3, 21 -- for {microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 14:30:58 -0400 3, 21 -- Message-ID: {4525798A.8090607-at-ufl.edu} 3, 21 -- Date: Thu, 05 Oct 2006 14:30:50 -0700 3, 21 -- From: greg Erdos {gwe-at-ufl.edu} 3, 21 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 3, 21 -- MIME-Version: 1.0 3, 21 -- To: microscopy-at-microscopy.com 3, 21 -- Subject: 2006 Tutorials Available 3, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 21 -- Content-Transfer-Encoding: 7bit 3, 21 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 21 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 3, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 3, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
I am interested in gathering advice from members of the list serve about lighting for our new FE TEM. We currently have a 3X3 grid of fluorescent lights for general lighting, and servicing the instrument etc. We've installed clean electrical service - back to the building ground with good wiring and checked for fields. In particular, I'd be interested in hearing from members about their experience with spot lighting installations to illuminate the TEM and workstations while in operation. In the past we have just installed track lighting with high intensity halogen spotlights mounted in movable heads to direct the light where we need the illumination. The spotlights were controlled with a dimmer switch to adjust the intensity to the desired level. I understand that mounting the dimmer switch to the microscope can lead to ground loop problems. What I'd like to hear is if anybody has experience with new installations, things to watch out for (ground loops, fields that effect the microscope etc.), and also any recomendations for lighting systems and or components. Thanks in advance,
Greg
-- -- ================================================================== Greg Strout Research Scientist, University of Oklahoma Past President, Oklahoma Microscopy Society (OMS) e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
==============================Original Headers============================== 6, 19 -- From gstrout-at-ou.edu Thu Oct 5 13:53:43 2006 6, 19 -- Received: from fourlom.ou.edu (mail.zero.ou.edu [129.15.0.75]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95IrhfC006506 6, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 13:53:43 -0500 6, 19 -- Received: from [127.0.0.1] ([129.15.38.202]) 6, 19 -- by fourlom.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 6, 19 -- 28 2005)) with ESMTP id {0J6O0050OF5G6W10-at-fourlom.ou.edu} for 6, 19 -- Microscopy-at-microscopy.com; Thu, 05 Oct 2006 13:53:41 -0500 (CDT) 6, 19 -- Date: Thu, 05 Oct 2006 13:53:37 -0500 6, 19 -- From: Greg Strout {gstrout-at-ou.edu} 6, 19 -- Subject: Room lighting and new microscope installation 6, 19 -- To: Microscopy-at-microscopy.com 6, 19 -- Message-id: {452554B1.40509-at-ou.edu} 6, 19 -- MIME-version: 1.0 6, 19 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-transfer-encoding: 7BIT 6, 19 -- X-Accept-Language: en-us, en 6, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 19 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
On Oct 5, 2006, at 11:53 AM, gstrout-at-ou.edu wrote:
} I am interested in gathering advice from members of the list serve } about } lighting for our new FE TEM. } We currently have a 3X3 grid of fluorescent lights for general } lighting, } and servicing the instrument etc. We've installed clean electrical } service - back to the building ground with good wiring and checked for } fields. } In particular, I'd be interested in hearing from members about their } experience with spot lighting installations to illuminate the TEM and } workstations while in operation. In the past we have just installed } track lighting with high intensity halogen spotlights mounted in } movable } heads to direct the light where we need the illumination. The } spotlights were controlled with a dimmer switch to adjust the intensity } to the desired level. } I understand that mounting the dimmer switch to the microscope can lead } to ground loop problems. } What I'd like to hear is if anybody has experience with new } installations, things to watch out for (ground loops, fields that } effect } the microscope etc.), and also any recomendations for lighting systems } and or components. } Dear Greg, We have two fixtures each with two 4' fluorescent lights and six track lighting fixtures with 50 W halogen flood lights. The fluorescents are controlled by either a wall switch or a switch mounted on the scope, and the halogens are controlled by a dimmer mounted on the scope--the wires to the switches are run through conduit along the scope frame. We have not seen any adverse effect from the scope-mounted switches on either our FEI T12 or FEI TF30H. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Thu Oct 5 15:05:13 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95K5DJn018514 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 5 Oct 2006 15:05:13 -0500 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 835D835456 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 5 Oct 2006 13:05:12 -0700 (PDT) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 420D210A0D4 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 5 Oct 2006 13:05:11 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200610051853.k95IrnNq006665-at-ns.microscopy.com} 4, 22 -- References: {200610051853.k95IrnNq006665-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {37d161280f0a3ff39290d89266cf039a-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Room lighting and new microscope installation 4, 22 -- Date: Thu, 5 Oct 2006 13:09:08 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
My lab is setup somewhat like yours, with fluorescents and incandescent track lighting.
I don't have a dimmer, but the track lighting is low brightness. When I need to see something under high light conditions, I use the fluorescents. To make this easy, I have a switch at the operating position (RF/wireless) to control the fluorescents. The track lights are on all the time.
Dimmers should not generate ground loops. However, most are pretty good at generating electrical noise that can be radiated, conveyed over the power line, or both. This may or may not be a problem depending on the dimmer quality, the lab layout and the susceptibility of the TEM. A noise free dimmer is a variable autotransformer (Variac TM). Nothing is perfect... They can generate magnetic fields. Fields tend to be small, however, due to the steel shielding and the torrodial core.
Another caveat... New high efficiency fluorescents use solid state switching devices instead of the old saturatable transformer "ballasts". If poorly designed, these can also generate electrical noise. FWIW, The ones I have (brand unknown) do not affect my SEM.
Regards, Woody BWXT Services
-----Original Message----- X-from: gstrout-at-ou.edu [mailto:gstrout-at-ou.edu] Sent: Thursday, October 05, 2006 2:54 PM To: White, Woody N.
Hi everyone,
I am interested in gathering advice from members of the list serve about
lighting for our new FE TEM. We currently have a 3X3 grid of fluorescent lights for general lighting,
and servicing the instrument etc. We've installed clean electrical service - back to the building ground with good wiring and checked for fields. In particular, I'd be interested in hearing from members about their experience with spot lighting installations to illuminate the TEM and workstations while in operation. In the past we have just installed track lighting with high intensity halogen spotlights mounted in movable
heads to direct the light where we need the illumination. The spotlights were controlled with a dimmer switch to adjust the intensity to the desired level. I understand that mounting the dimmer switch to the microscope can lead to ground loop problems. What I'd like to hear is if anybody has experience with new installations, things to watch out for (ground loops, fields that effect
the microscope etc.), and also any recomendations for lighting systems and or components. Thanks in advance,
Greg
-- -- ================================================================== Greg Strout Research Scientist, University of Oklahoma Past President, Oklahoma Microscopy Society (OMS) e-mail: gstrout-at-ou.edu Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
==============================Original Headers============================== 6, 19 -- From gstrout-at-ou.edu Thu Oct 5 13:53:43 2006 6, 19 -- Received: from fourlom.ou.edu (mail.zero.ou.edu [129.15.0.75]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95IrhfC006506 6, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 5 Oct 2006 13:53:43 -0500 6, 19 -- Received: from [127.0.0.1] ([129.15.38.202]) 6, 19 -- by fourlom.ou.edu (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 6, 19 -- 28 2005)) with ESMTP id {0J6O0050OF5G6W10-at-fourlom.ou.edu} for 6, 19 -- Microscopy-at-microscopy.com; Thu, 05 Oct 2006 13:53:41 -0500 (CDT) 6, 19 -- Date: Thu, 05 Oct 2006 13:53:37 -0500 6, 19 -- From: Greg Strout {gstrout-at-ou.edu} 6, 19 -- Subject: Room lighting and new microscope installation 6, 19 -- To: Microscopy-at-microscopy.com 6, 19 -- Message-id: {452554B1.40509-at-ou.edu} 6, 19 -- MIME-version: 1.0 6, 19 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 6, 19 -- Content-transfer-encoding: 7BIT 6, 19 -- X-Accept-Language: en-us, en 6, 19 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 6, 19 -- Gecko/20040804 Netscape/7.2 (ax) ==============================End of - Headers==============================
==============================Original Headers============================== 24, 26 -- From nwwhite-at-bwxt.com Thu Oct 5 15:15:51 2006 24, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 24, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k95KFo7O029126 24, 26 -- for {microscopy-at-msa.microscopy.com} ; Thu, 5 Oct 2006 15:15:51 -0500 24, 26 -- Received: from ([131.184.13.224]) 24, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.2375910; 24, 26 -- Thu, 05 Oct 2006 16:15:32 -0400 24, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 24, 26 -- Thu, 5 Oct 2006 16:15:32 -0400 24, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 24, 26 -- Content-class: urn:content-classes:message 24, 26 -- MIME-Version: 1.0 24, 26 -- Content-Type: text/plain; 24, 26 -- charset="US-ASCII" 24, 26 -- Subject: RE: [Microscopy] Room lighting and new microscope installation 24, 26 -- Date: Thu, 5 Oct 2006 16:15:32 -0400 24, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D098-at-BWXSPO01.BWXS.BWXTECH.NET} 24, 26 -- X-MS-Has-Attach: 24, 26 -- X-MS-TNEF-Correlator: 24, 26 -- Thread-Topic: [Microscopy] Room lighting and new microscope installation 24, 26 -- Thread-Index: Acbor7FTkYpsjxQGQq6Azv3EOIESJgAB+2ow 24, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 24, 26 -- To: {gstrout-at-ou.edu} , "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 24, 26 -- X-OriginalArrivalTime: 05 Oct 2006 20:15:32.0570 (UTC) FILETIME=[028903A0:01C6E8BB] 24, 26 -- Content-Transfer-Encoding: 8bit 24, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k95KFo7O029126 ==============================End of - Headers==============================
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 ngohad-at-clemson.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: ngohad-at-clemson.edu Name: Neeraj V. Gohad
Organization: Dept. Biological Sciences, Clemson University
Title-Subject: [Filtered] Confocal Microscopy
Question: Dear All,
I am going to purchase some DVDs and Tapes from MSA on various topics in microscopy. Could any one please recommend a DVD or tape from MSAs list on Confocal microscopy. There are a buch on confocal microscopy. I am getting to be an advanced user so I want something that will lecture on advanced techniques.
Please let me know,
Thank you in advance,
Neeraj V. Gohad Graduate Research Assistant, Dept. Biological Sciences, Clemson University.
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 dang-at-oakland.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: dang-at-oakland.edu Name: Loan Dang
Organization: Oakland University
Title-Subject: [Filtered] The protocol for fixing the plant sample
Question: I would like to have the standard procedure for fixing the plant material. I need to fix the maple leaf sample. Thanks LD
Hello, I am so very grateful to all who send me advices and hints how to solve our trouble with the knifemaker. Our LKB 7801A is now working fine. Thanking you, again.
My best regards from Prague Oldrich
==============================Original Headers============================== 2, 20 -- From benada-at-biomed.cas.cz Fri Oct 6 01:43:19 2006 2, 20 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 2, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k966hI6o007213 2, 20 -- for {microscopy-at-microscopy.com} ; Fri, 6 Oct 2006 01:43:18 -0500 2, 20 -- Received: from [147.231.44.101] (u117ob.mbu.cas.cz [147.231.44.101]) 2, 20 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id k966dBI0009934 2, 20 -- for {microscopy-at-microscopy.com} ; Fri, 6 Oct 2006 08:39:11 +0200 (CEST) 2, 20 -- From: benada-at-biomed.cas.cz 2, 20 -- To: microscopy-at-microscopy.com 2, 20 -- Date: Fri, 06 Oct 2006 08:43:14 +0200 2, 20 -- MIME-Version: 1.0 2, 20 -- Subject: LKB knifemaker 7801A problem - Thanks 2, 20 -- Message-ID: {45261722.18601.161A14-at-benada.biomed.cas.cz} 2, 20 -- Priority: normal 2, 20 -- X-mailer: Pegasus Mail for Windows (4.41) 2, 20 -- Content-type: text/plain; charset=US-ASCII 2, 20 -- Content-transfer-encoding: 7BIT 2, 20 -- Content-description: Mail message body 2, 20 -- X-Antivirus: avast! (VPS 0640-3, 05.10.2006), Outbound message 2, 20 -- X-Antivirus-Status: Clean ==============================End of - Headers==============================
Thanks to all those who replied to my query regarding the sectioning of cotton fibres for TEM. As usual the replies were exceptionally useful and have given us a number of leads to follow.
Cheers and have a good weekend.
Colin Veitch
Electron Microscopist CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia. E-mail: colin.veitch-at-csiro.au Web: http://www.tft.csiro.au
The information contained in this e-mail message may be privileged or confidential information. If you are not an intended recipient, you may not copy, distribute or take any action in reliance on it. If you have received this message in error, please telephone CSIRO Textile and Fibre Technology on +61 3 5246 4000.
==============================Original Headers============================== 8, 28 -- From Colin.Veitch-at-csiro.au Fri Oct 6 01:52:44 2006 8, 28 -- Received: from vic-MTAout2.csiro.au (vic-MTAout2.csiro.au [150.229.64.38]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k966qhA9017786 8, 28 -- for {microscopy-at-msa.microscopy.com} ; Fri, 6 Oct 2006 01:52:44 -0500 8, 28 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=MJfZXFYeG9il5Dear5zfD95j+OvuR3g38phqv+vcGJkMUpCIOxR6YB1ge+vn9V59t7fqCU9/UciOFHugqzo8ip/BSkUOsGZNniCYxEoQIUB47oIZ1BUrI/YiwCUVqquE; 8, 28 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 8, 28 -- by vic-MTAout2.csiro.au with ESMTP; 06 Oct 2006 16:52:43 +1000 8, 28 -- X-IronPort-AV: i="4.09,268,1157292000"; 8, 28 -- d="scan'208"; a="97313713:sNHT25809548" 8, 28 -- Received: from exvic5-gex.nexus.csiro.au ([138.194.194.6]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 8, 28 -- Fri, 6 Oct 2006 16:52:36 +1000 8, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 28 -- content-class: urn:content-classes:message 8, 28 -- MIME-Version: 1.0 8, 28 -- Content-Type: text/plain; 8, 28 -- charset="us-ascii" 8, 28 -- Subject: Cotton sectioning thanks 8, 28 -- Date: Fri, 6 Oct 2006 16:52:35 +1000 8, 28 -- Message-ID: {32CDDDAA7161394599F00254949157490248E3-at-exvic5-gex.nexus.csiro.au} 8, 28 -- X-MS-Has-Attach: 8, 28 -- X-MS-TNEF-Correlator: 8, 28 -- Thread-Topic: Cotton sectioning thanks 8, 28 -- Thread-Index: AcbpE/3rE6/8kEF8QQWwfZ6SLEINnw== 8, 28 -- From: {Colin.Veitch-at-csiro.au} 8, 28 -- To: {microscopy-at-msa.microscopy.com} 8, 28 -- X-OriginalArrivalTime: 06 Oct 2006 06:52:36.0202 (UTC) FILETIME=[019864A0:01C6E914] 8, 28 -- Content-Transfer-Encoding: 8bit 8, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k966qhA9017786 ==============================End of - Headers==============================
Hi Dang, Your question is impossible to answer without more information. What do you want to observe in the plant, and with what type of microscopy? The choice of protocol is guided by the answers to those questions.
By the way, to find a lot of information about this in a small space, search your library catalog for "microtechnique". That word sounds like it means delicate surgery or something but in fact it is what books about fixation protocols are often titled. There probably are one or two at your library (try Plant or Botanical Microtechnique).
Good luck, Tobias
} Organization: Oakland University } } Title-Subject: [Filtered] The protocol for fixing the plant sample } } Question: I would like to have the standard procedure for fixing the } plant material. } I need to fix the maple leaf sample. } Thanks } LD } } -
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (brookie_cookie0-at-hotmai.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Thursday, October 5, 2006 at 09:19:36 ---------------------------------------------------------------------------
Looking for methods or experiences in measuring the total surface area of 1 gram of mineral (diopside) powder with grain sizes between 150 and 250 microns. I have heard of SEM techniques but not sure how that is done. Any Ideas?
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-smu.edu
==============================Original Headers============================== 6, 23 -- From rbeavers-at-mail.smu.edu Fri Oct 6 15:06:07 2006 6, 23 -- Received: from s31xua.systems.smu.edu (s31xua.systems.smu.edu [129.119.70.72]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k96K66Gd007881 6, 23 -- for {Microscopy-at-microscopy.com} ; Fri, 6 Oct 2006 15:06:07 -0500 6, 23 -- Received: from s31xe7.systems.smu.edu ([129.119.70.140]) by s31xua.systems.smu.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Fri, 6 Oct 2006 15:06:04 -0500 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="us-ascii" 6, 23 -- Subject: Measurement of surface area 6, 23 -- Date: Fri, 6 Oct 2006 15:06:04 -0500 6, 23 -- Message-ID: {F8AF6BF6CD1CC040AF35B0C2D1680BBB235E26-at-s31xe7.systems.smu.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: Measurement of surface area 6, 23 -- Thread-Index: AcbpgtoCt0jPMxFLTnOwa3b2evFx2g== 6, 23 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} 6, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 06 Oct 2006 20:06:05.0119 (UTC) FILETIME=[DAB89CF0:01C6E982] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k96K66Gd007881 ==============================End of - Headers==============================
The University of Wisconsin Milwaukee is searching for a full time probationary senior instrumentation specialist to train and assist faculty, research and technical staff and students in use of electron and atomic force microscopes, vacuum systems, and specimen preparation instruments; to install, operate, update, maintain and repair scientific instruments; to administer instrument use and enforce lab and chemical safety protocols. Candidates must have a Bachelor's degree with course work in physical or biological sciences, a post-bachelors certificate or degree in electron microscopy; at least four years of experience in microscopy and /or surface science; ability to effectively operate research instruments and computers; strong written and verbal communication skills to work with lab users from diverse backgrounds. Well qualified applicants will have training and/or experience with HRTEM, AFM, SQUID, vacuum leak checkers, preparation of surface and TEM specimen. Minimum salary of $46,191, commensurate with qualifications. UWM is an AA/EEO employer
To apply, send letter of intent and resume with three references, postmarked by Nov. 8, 2006, to: Search and Screen Chair - Microscopy, UWM L&S Holton 249, P. O. Box 413, Milwaukee, WI 53201-0413, For more information please see www.uwm.edu/letsci/jobs
Marija Gajdardziska-Josifovska Associate Dean for Natural Sciences University of Wisconsin Milwaukee tel: 414 229 2925 fax 414 229 6827
==============================Original Headers============================== 4, 24 -- From marijagj-at-gmail.com Fri Oct 6 16:49:53 2006 4, 24 -- Received: from nf-out-0910.google.com (nf-out-0910.google.com [64.233.182.188]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k96Lnq7j019806 4, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 6 Oct 2006 16:49:53 -0500 4, 24 -- Received: by nf-out-0910.google.com with SMTP id c2so848197nfe 4, 24 -- for {Microscopy-at-Microscopy.Com} ; Fri, 06 Oct 2006 14:49:51 -0700 (PDT) 4, 24 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 24 -- s=beta; d=gmail.com; 4, 24 -- h=received:message-id:date:from:to:subject:cc:mime-version:content-type:content-transfer-encoding:content-disposition; 4, 24 -- b=rs8qh+vc/Pr73QSxeHHY9YaCfsiYJHuwU0K+8cHSLdjrdl1I6kCv15/14qIPaKlYHzPVnIi9tPTs2wiZGkL3wiu6WInnpSXY5Umpbz9cPBlF57zN55x61zkoLykCpo4UugIymVRRqCX9dFwPvTNMUT2wTaTIVyRIrPzD1lihg4M= 4, 24 -- Received: by 10.49.8.1 with SMTP id l1mr6060288nfi; 4, 24 -- Fri, 06 Oct 2006 14:49:51 -0700 (PDT) 4, 24 -- Received: by 10.49.87.12 with HTTP; Fri, 6 Oct 2006 14:49:51 -0700 (PDT) 4, 24 -- Message-ID: {ee4af5560610061449x27fa2469x25e6a2d6292e058b-at-mail.gmail.com} 4, 24 -- Date: Fri, 6 Oct 2006 16:49:51 -0500 4, 24 -- From: "Marija Gajdardziska-Josifovska" {marijagj-at-gmail.com} 4, 24 -- To: Microscopy-at-Microscopy.Com 4, 24 -- Subject: Senior Instrumentation Specialist Position 4, 24 -- Cc: "Linda Daley" {lindaley-at-uwm.edu} , 4, 24 -- "Marija Gajdardziska-Josifovska" {mgj-at-uwm.edu} 4, 24 -- MIME-Version: 1.0 4, 24 -- Content-Type: text/plain; charset=UTF-8; format=flowed 4, 24 -- Content-Transfer-Encoding: 7bit 4, 24 -- Content-Disposition: inline ==============================End of - Headers==============================
Version 2.0.0 of the 2dx software for image-processing for 2D crystals is now available here: http://2dx.org/download/2dx-software
This version contains numerous new features, and has included all the bugfixes, suggestions, and better algorithms that were brought to us from several users.
A 13-minute movie that demonstrates the basic usage of 2dx is available here: http://2dx.org/documentation/2dx-software/tutorial/2dx_image-primer
A description of 2dx in JSB can be found here: http://dx.doi.org/10.1016/j.jsb.2006.07.020
Bryant, Xiangyan and Henning.
Henning Stahlberg, Molecular & Cellular Biology, Briggs Hall 5, University of California at Davis, 1 Shields Ave., Davis, CA 95616, USA Tel: +1-530-752 8282 (office), +1-530-754 8285 (lab) Fax: +1-530-752 3085 mailto:HStahlberg-at-ucdavis.edu http://stahlberglab.ucdavis.edu
==============================Original Headers============================== 11, 18 -- From HStahlberg-at-ucdavis.edu Fri Oct 6 19:15:05 2006 11, 18 -- Received: from pop23.ucdavis.edu (pop23.ucdavis.edu [169.237.105.13]) 11, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k970F4kc032124 11, 18 -- for {Microscopy-at-microscopy.com} ; Fri, 6 Oct 2006 19:15:05 -0500 11, 18 -- Received: from [169.237.214.65] ([169.237.214.65]) 11, 18 -- by pop23.ucdavis.edu (8.13.7/8.13.1/it-std-5.2.0) with ESMTP id k970Dsxk000955; 11, 18 -- Fri, 6 Oct 2006 17:15:04 -0700 (PDT) 11, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) 11, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 11, 18 -- Message-Id: {D71030DD-B067-4DBE-8B42-B998585F0F94-at-ucdavis.edu} 11, 18 -- Cc: "Bryant R. Gipson" {BRGipson-at-ucdavis.edu} , 11, 18 -- Xiangyan Zeng {XZeng-at-ucdavis.edu} 11, 18 -- Content-Transfer-Encoding: 7bit 11, 18 -- From: Henning Stahlberg {HStahlberg-at-ucdavis.edu} 11, 18 -- Subject: 2dx - Image Processing for 2D Crystal Images: Version 2.0.0 now available 11, 18 -- Date: Fri, 6 Oct 2006 17:15:04 -0700 11, 18 -- To: Microscopy-at-microscopy.com 11, 18 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (JawaharMKD-at-aol.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, October 6, 2006 at 17:33:58 ---------------------------------------------------------------------------
Email: JawaharMKD-at-aol.com Name: Sonia
Organization: Queen Mary
Education: Undergraduate College
Location: London, UK
Question: What is the mechanism for simple staining? How does Gram staining actually work? What is the principle behind streaking on a plate? Why do we use selective or differential media?
} Just wanted to ask for your recommendation of a good } brand/model of UPS for an SEM system. } } We have a Phillips/FEI Quanta 200 ESEM and we are thinking of } buying an uninterrupted power supply for voltage surge protection.
Our Quanta 400 is getting along well with a Powerware 9125 6000VA. I sometimes wish it offered a little more headroom because the initial pumpdown sequence will infrequently trigger an overload warning, but under normal operating conditions, 24/7, it is fine. Powerware support is excellent. They replaced a UPS that was frequently giving me overload warnings with a UPS that subsequently hasn't issued a warning yet. Ours also supports 2 computers along with an EDX system, and if your installation is like mine, there's no need to support a chiller, so the same KVA may be all you need.
It is also the case that our UPS, with only a pair of batteries, is only intended to save power for relatively short 15 minute outages, which covers ~99% of our problems. If you want extended operation, then Powerware offers additional battery modules for this same model.
A UPS this size can be noisy, and possibly emit detrimental EM radiation. Ours is in a different room along with the 208-to-230V step-up transformer. 230V is the input voltage for the UPS.
See: http://www.powerware.com/UPS/9125RM_UPS.asp
HTH, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre c/o Memorial University St. John's, NL A1C 5S7
==============================Original Headers============================== 10, 20 -- From michael-at-shaffer.net Mon Oct 9 08:48:56 2006 10, 20 -- Received: from n007.sc1.cp.net (smtpout1453.sc1.he.tucows.com [64.97.157.153]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k99DmtTb027557 10, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 9 Oct 2006 08:48:55 -0500 10, 20 -- Received: from rarewolf (205.251.84.119) by n007.sc1.cp.net (7.2.069.1) (authenticated as michael-at-shaffer.net) 10, 20 -- id 452428650012157A; Mon, 9 Oct 2006 13:48:50 +0000 10, 20 -- From: "michael shaffer" {michael-at-shaffer.net} 10, 20 -- To: {mmiralles-at-pi.ac.ae} 10, 20 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 10, 20 -- Subject: RE: [Microscopy] SEM : UPS model for Fei E-SEM Quanta 200 10, 20 -- Date: Mon, 9 Oct 2006 11:18:52 -0230 10, 20 -- Message-ID: {009201c6eba9$a8b8c6c0$4701a8c0-at-rarewolf} 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; 10, 20 -- charset="US-ASCII" 10, 20 -- Content-Transfer-Encoding: 7bit 10, 20 -- X-Mailer: Microsoft Office Outlook 11 10, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 10, 20 -- Thread-Index: AcbrhcqqoOqJZ/dISOGHCQh0ezXlKAAIRzQQ 10, 20 -- In-Reply-To: {200610090932.k999Wind013147-at-ns.microscopy.com} ==============================End of - Headers==============================
If your concern is surges or sags in power supply, you don't need an UPS. What you would need is a stabilizer like Topaz makes. This is a magnetic resonance system that adjusts output voltage to be at specified output. It also includes input spike protection and EMI/RFI removal.
If you do want an UPS, and these are very nice to have, there are several makers. I use two Liebert Nfinity 8KVA units with redundant power and control modules. SEM and its PC are on one UPS while EDS, EBSD and other PCs and chiller and compressor are on another one. Each UPS runs at about 25% total capacity and will run the whole system for about 250 minutes if power fails--which it usually does in winter and summer here in California. These units regulate output voltage to within about 2% of specified value. They are double conversion in that they take in AC, convert it to DC and invert the DC to AC output. When input AC fails, DC from the batteries is inverted to AC output. Along with this is spike protection, sag protection and EMI/RFI protection.
These units work on 208-240VAC input and output similar voltage. I take 240VAC in and output 240VAC and 120VAC.
Hope this helps.
gary g.
At 02:35 AM 10/9/2006, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 12, 20 -- From gary-at-gaugler.com Mon Oct 9 12:35:58 2006 12, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k99HZwTn009964 12, 20 -- for {microscopy-at-microscopy.com} ; Mon, 9 Oct 2006 12:35:58 -0500 12, 20 -- Received: (qmail 9350 invoked from network); 9 Oct 2006 10:35:57 -0700 12, 20 -- Received: by simscan 1.1.0 ppid: 9347, pid: 9348, t: 0.1485s 12, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 20 -- by qsmtp2 with SMTP; 9 Oct 2006 10:35:57 -0700 12, 20 -- Message-Id: {7.0.1.0.2.20061009102243.025c3fd8-at-gaugler.com} 12, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 20 -- Date: Mon, 09 Oct 2006 10:35:59 -0700 12, 20 -- To: mmiralles-at-pi.ac.ae 12, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 20 -- Subject: Re: [Microscopy] SEM : UPS model for Fei E-SEM Quanta 200 12, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 20 -- In-Reply-To: {200610090935.k999ZeJY015446-at-ns.microscopy.com} 12, 20 -- References: {200610090935.k999ZeJY015446-at-ns.microscopy.com} 12, 20 -- Mime-Version: 1.0 12, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
David, sorry for my late answer; I had been on holidays in Italy :-)))
The DSMs are very nice instruments but from what I know a little complicated in trouble-shooting. If you need some original service manuals I can try and copy these from a friend of mine.
Yes, definitely there should be more high-tech available at high-schools or some joint-ventures between high-schools and universities to share possibilities...
In a kind of way I am running out of "service-time" adopting more than the existing two schools, but - thinking of all those field-engineers out there from different manufacturers someone at FEI, HITACHI, JEOL or ZEISS NTS should think about doing some adoption of schools...
Modells for integration within the school system very largely depend on the teachers there (my experience) and your time available for coaching kids and teachers and doing updates sometimes a year. Get the machines known throughout the school, do some nice image and put it on floor walls....
----- Original Message ----- X-from: "David L. Jones" {dljones-at-bestweb.net} To: {stefan.diller-at-t-online.de} Sent: Thursday, September 28, 2006 6:45 PM
I was very interested in the several different ways people have developed for back-filling SEM specimen chambers with dry nitrogen gas. I think, however, that years ago we developed a method that is even simpler and less expensive than any of those described so far. This method is described in detail on page 65 of my book Vacuum Methods in Electron Microscopy (available from SPI Supplies, M. E. Taylor, Ladd, etc.)
What we did was to run a flexible hose (ordinary polyethylene tubing does just fine) from a stopper in the mouth of the Dewar flask of an EDS detector (you could alternatively connect to the bleed-off line on a liquid nitrogen storage tank) to the gas inlet on the SEM's specimen chamber. A large flexible plastic container, such as an inflatable plastic toy or beach ball, was connected to a T-junction in this line by means of a piece of soft, flexible surgical rubber tubing, and a straight, clean slit about 100 mm long was made in this tubing with a sharp razor blade or scalpel. This slit will normally stay closed tightly enough so that the nitrogen boiling off from the liquid nitrogen container will flow into the plastic ball. When the ball becomes full, the slit will open enough to prevent the system from becoming over pressurized, thus serving as a primitive pressure relief valve. When the inlet valve to the specimen chamber is opened, the plastic ball will collapse and the nitrogen will flow into the specimen chamber, but only under atmospheric pressure so there will be no danger of over pressurization. A small weight can be placed on the plastic ball to maintain a small flow of nitrogen after the system reaches atmospheric pressure, if desired. A beach ball about 0.5 m in diameter will hold enough nitrogen to fill most vacuum chambers several times, and it turns out to be lots of fun explaining to visitors why you have the ball connected to your electron microscope.
-- Wilbur C. Bigelow, Professor Emeritus Materials Sci. & Engr., Univ. of Michigan Ann Arbor, Michigan 48109-2136 e-mail: bigelow-at-engin.umich.edu; Fx:734-763-4788; Ph:734-975-0858 Address mail to: 2911 Whittier Court Ann Arbor, MI 48104-6731
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Email: jonas-baltrusaitis-at-uiowa.edu Name: Jonas Baltrusaitis
Organization: Central Microscopy Research Facility, Univ. of Iowa
Title-Subject: [Filtered] SEC from FESEM
Question: All,
Where can I purchase an old SEM specimen exchange chamber? I want to modify it and use as a quick load chamber on my existing Kratos AXIS Ultra XPS system.
RESEARCH ASSOCIATE 3 OR 4 (Microscopy Imaging Specialist) Department of Biological Sciences
The Socolofsky Microscopy Facility at LSU (http://www.biology.lsu.edu/facilities/micro_fac/) has state of the art light microscopy equipment, including a Leica TCS SP2 confocal system, deconvolution imaging system based on a Leica DM RXA2 upright, and a newly acquired Zeiss Lumar.V12 fluorescence stereomicroscope. Required Qualifications: (Research Associate 3): Bachelor's or equivalent degree in Biological Science or related field with three years relevant experience OR a Master's degree with one year experience. (Research Associate 4): Master's degree with two years relevant experience OR Ph.D. Experience in confocal light microscopy is REQUIRED for either level (RA3 or RA4). NOTE: This announcement is for ONE position at either RA3 or RA4 level, not for two positions.
Responsibilities: images acquisition and analysis, user training, and oversight of light microscopes and multi-user digital imaging equipment. An offer of employment is contingent on a satisfactory pre-employment background check. Application deadline is November 10, 2006, or until candidate is selected.
Submit letter of application and resume (including email address), and two letters of recommendation to:
Ms. Charyl Thompson Dept. of Biological Sciences 202 Life Sciences Bldg. Louisiana State University Ref: #014225 Baton Rouge, LA 70803. E-mail: cthomps-at-lsu.edu
LSU IS AN EQUAL OPPORTUNITY/EQUAL ACCESS EMPLOYER
_______________________________________________
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==============================Original Headers============================== 10, 20 -- From dburk-at-lsu.edu Tue Oct 10 09:20:52 2006 10, 20 -- Received: from gate011.lsu.edu (gate011.ocs.lsu.edu [130.39.184.213]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9AEKpiG006232 10, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 10 Oct 2006 09:20:51 -0500 10, 20 -- Received: from smc30 ([130.39.115.181]) 10, 20 -- by gate011.lsu.edu (Lotus Domino Release 6.0.3) 10, 20 -- with ESMTP id 2006101009205099-1197 ; 10, 20 -- Tue, 10 Oct 2006 09:20:50 -0500 10, 20 -- From: "David Burk" {dburk-at-lsu.edu} 10, 20 -- To: {Microscopy-at-microscopy.com} 10, 20 -- Subject: Position Available - Microscopy Imaging Specialist 10, 20 -- Date: Tue, 10 Oct 2006 09:20:48 -0500 10, 20 -- Message-ID: {000c01c6ec77$483d0620$b5732782-at-biosc.lsu.edu} 10, 20 -- MIME-Version: 1.0 10, 20 -- X-Mailer: Microsoft Office Outlook 11 10, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 10, 20 -- Thread-Index: Acbsd0guMJVzZ0wWSPKoCUaPciQA8g== 10, 20 -- Content-Transfer-Encoding: 7bit 10, 20 -- Content-Type: text/plain; 10, 20 -- charset="us-ascii" ==============================End of - Headers==============================
I don't see Topaz much either. I replaced the Topaz unit several years ago and do not have the model number here any longer. It probably got replaced by a new one or the line shut down. What you need is a ferroresonant transformer like:
These are USA links but probably TSI or Sola have international outlets.
If you check the data on the links, these are probably what you seek rather than an UPS. If power is on all the time but dirty, the ferroresonant units are the ticket. They can be much cheaper than an UPS of same capacity. Plus, they easily can handle three phase input power if you have it.
gary g.
At 02:07 AM 10/10/2006, you wrote:
} Hi gary, } } Can you give me a more specific model of this Topaz stabilizer? I only } saw a single model at the web and it didn't post the technical details. } } Thanks again, } Melina } } } -----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] } Sent: Monday, October 09, 2006 9:36 PM } To: Melina Miralles } Cc: MSA listserver } Subject: Re: [Microscopy] SEM : UPS model for Fei E-SEM Quanta 200 } } If your concern is surges or sags in power supply, you } don't need an UPS. What you would need is a stabilizer } like Topaz makes. This is a magnetic resonance system } that adjusts output voltage to be at specified output. } It also includes input spike protection and EMI/RFI } removal. } } If you do want an UPS, and these are very nice to have, } there are several makers. I use two Liebert Nfinity } 8KVA units with redundant power and control modules. } SEM and its PC are on one UPS while EDS, EBSD and other } PCs and chiller and compressor are on another one. Each } UPS runs at about 25% total capacity and will run the } whole system for about 250 minutes if power fails--which } it usually does in winter and summer here in California. } These units regulate output voltage to within about 2% of } specified value. They are double conversion in that they } take in AC, convert it to DC and invert the DC to AC output. } When input AC fails, DC from the batteries is inverted to AC } output. Along with this is spike protection, sag protection } and EMI/RFI protection. } } These units work on 208-240VAC input and output similar } voltage. I take 240VAC in and output 240VAC and 120VAC. } } Hope this helps. } } gary g.
==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Tue Oct 10 18:20:24 2006 11, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9ANKOjQ028072 11, 20 -- for {microscopy-at-microscopy.com} ; Tue, 10 Oct 2006 18:20:24 -0500 11, 20 -- Received: (qmail 14657 invoked from network); 10 Oct 2006 16:17:22 -0700 11, 20 -- Received: by simscan 1.1.0 ppid: 14652, pid: 14654, t: 0.1668s 11, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 20 -- by qsmtp4 with SMTP; 10 Oct 2006 16:17:22 -0700 11, 20 -- Message-Id: {7.0.1.0.2.20061010161328.0257cd40-at-gaugler.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 20 -- Date: Tue, 10 Oct 2006 16:20:25 -0700 11, 20 -- To: "Melina Miralles" {mmiralles-at-pi.ac.ae} 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: RE: [Microscopy] SEM : UPS model for Fei E-SEM Quanta 200 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {D5603421C6303A46883F87E7968F285B01DEAA28-at-pi-exm.PI.AC.AE} 11, 20 -- References: {D5603421C6303A46883F87E7968F285B01DEAA28-at-pi-exm.PI.AC.AE} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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Email: robertoclark-at-gmail.com Name: Rob Clark
Organization: Sharp HealthCare
Title-Subject: [Filtered] Decacifying teeth
Question: Hi All,
I am debating whether or not to decalcify a root of a tooth in a formic acid/formaldehyde decal such as CalRite, or to go the slow route with EDTA/glut. The root will then be processed for TEM studies in hopes to identify microorganisms. No other studies are planned. From your experience will formic acid be the way to go or will it compromise my results?
Thank you so much in advance. Rob Clark E.M. Technologist Sharp HealthCare San Diego CA
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Email: ppq3k-at-yahoo.com Name: Peng Li
Organization: University of New Mecico
Title-Subject: [Filtered] Electron Microscopist position at Univ. of New Mexico
Question: Research Scientist 1/Electron Microscopist Department of Earth and Planetary Sciences, University of New Mexico
The University of New Mexico at UNM is seeking applicants for a Research Scientist 1/Electron Microscopist for the Transmission Electron Microscope Laboratory in the Department of Earth and Planetary Sciences. The laboratory is a state of the art multiuser facility that includes JEOL 2010 high resolution TEM and JEOL 2010F FASTEM FEG TEM/STEM with an Oxford ISIS EDS and GATAN GIF system, and sample preparation facilities including ion mills, ultramicrotome and carbon coaters. The Research Technologist will be responsible for the day to day running of the laboratory in close collaboration with the laboratory manager, including training and assisting student, staff and faculty users, purchasing of laboratory consumables, sample preparation, laboratory safety, billing and record keeping. The successful candidate will also be expected to maintain and troubleshoot laboratory and TEM equipment and supervise service engineers. Full details of the position and application procedure can be obtained at the website address: http://jobs.unm.edu/jobopenings.cfm. Closing date for the position is October 30, 2006. For additional information contact Professor Adrian Brearley (Tel: 505 277 4163, e-mail :brearley-at-unm.edu) or Dr Peng Li (Tel: 505 277 7536; e-mail: pengli-at-unm.edu).
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Title-Subject: [Filtered] An Free Kevex Light Element Detector
Question: We have an 15 year old Kevex detector (light elements, Be window) we are pitching. Is anyone interested? I still have the shipping box. You might have to pay for the shipping???? I'd have to ask TPTB.
Perhaps we could exchange our views about the optimal buffers to use during fixations for TEM. Cacodylate is a traditional buffer, but it is not really healthy and perhaps it is time to update our knowledge. I have read that PIPES offered the lowest extraction but on the downside a less effecient buffering. Phosphate buffer does not allow the use of Ca and Mg, which could help stabilize some structures. HEPES buffer...well I don't know the downside, perhaps it offers only advantages ;-)but it is used pretty scarcely by the electron microscopists. Some other parameters can be taken into account: the storage, bench (or fridge) life, morphology {} immunology, plants {} animals cells and so on.
Lets talk about salts!
Personally I use Cacodylate for classical fixation of cells for morphology because it has also been used in the laboratory but I am eager to change. There is certainly some room for improvements in the morphology (especially of the membranes). HEPES seems to be a good candidate.
Stephane
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This Question was submitted to Ask-A-Microscopist by (winnie.wino-at-gmail.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, October 11, 2006 at 04:28:32 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both winnie.wino-at-gmail.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: winnie.wino-at-gmail.com Name: Winnie
Organization: NUS
Education: Undergraduate College
Location: Singapore
Title: Sample preparation
Question: Dear Microscopist, I have a few specimens (Charcoal and virgin wood) of which the sizes are 100x100x50(mm) and 30x30x100(mm). What should I prepare for those specimens before bringing them to the Lab for SEM scanning? Can the SEM machine's chamber accomodate such specimens? Should I reduce the size? Thank you and Best Regards. Winnie
Buffers have been on my mind lately too. In this laboratory, we have switched our buffer system over to Dulbecco's DMEM cell culture medium. In large part this is a matter of convenience. We do quite a few immunocytochemistry experiments, labeling culture cell membranes with a variety of antibodies, and we dilute these antibodies and subsequent colloidal gold conjugates with DMEM. The cells remain viable during incubations in antibody and secondary conjugates. It seemed reasonable to us to continue with DMEM during fixation, and we therefore dilute our cacodylate buffered 3%glut/3%paraformaldehyde stock solution with DMEM and also our OsO4 in DMEM. We have been pleased with the quality of the fixation and now we also use DMEM for buffering fixatives of tissues with equally pleasing results. But I have to admit to a nagging feeling that I am breaking some kind of microscopy law and would appreciate comments.
Thanks,
Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Wednesday, October 11, 2006 1:15 AM To: drk-at-shcc.org
Wealthy colleagues (yes: knowledge is a wealth!),
Perhaps we could exchange our views about the optimal buffers to use during fixations for TEM. Cacodylate is a traditional buffer, but it is not really healthy and perhaps it is time to update our knowledge. I have read that PIPES offered the lowest extraction but on the downside a less effecient buffering. Phosphate buffer does not allow the use of Ca and Mg, which could help stabilize some structures. HEPES buffer...well I don't know the downside, perhaps it offers only advantages ;-)but it is used pretty scarcely by the electron microscopists. Some other parameters can be taken into account: the storage, bench (or fridge) life, morphology {} immunology, plants {} animals cells and so on.
Lets talk about salts!
Personally I use Cacodylate for classical fixation of cells for morphology because it has also been used in the laboratory but I am eager to change. There is certainly some room for improvements in the morphology (especially of the membranes). HEPES seems to be a good candidate.
Stephane
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==============================Original Headers============================== 17, 21 -- From drk-at-shcc.org Wed Oct 11 10:19:56 2006 17, 21 -- Received: from SUNNIE.shcc.org (mail.shcc.org [64.213.211.202]) 17, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BFJtsx013068 17, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 10:19:55 -0500 17, 21 -- Received: from DRK2 ([64.213.211.62]) by shcc.org (PMDF V6.3 #31278) 17, 21 -- with ESMTP id {0J6Z00E0V8V2X5-at-shcc.org} for Microscopy-at-Microscopy.Com; Wed, 17, 21 -- 11 Oct 2006 08:11:26 -0700 (PDT) 17, 21 -- Date: Wed, 11 Oct 2006 08:19:49 -0700 17, 21 -- From: Doug Keene {drk-at-shcc.org} 17, 21 -- Subject: RE: [Microscopy] debating about buffers for TEM fixation 17, 21 -- In-reply-to: {200610110814.k9B8EqC0024591-at-ns.microscopy.com} 17, 21 -- To: Microscopy-at-Microscopy.Com 17, 21 -- Reply-to: drk-at-shcc.org 17, 21 -- Message-id: {0J6Z00E0W8V2X5-at-shcc.org} 17, 21 -- Organization: Shriners Hospitals for Children 17, 21 -- MIME-version: 1.0 17, 21 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 17, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 17, 21 -- Content-type: text/plain; charset=us-ascii 17, 21 -- Content-transfer-encoding: 7bit 17, 21 -- Thread-Index: AcbtDChpSOQbSAYeQd2UYXPEktNGrQAOl+9g ==============================End of - Headers==============================
For 40 years we have used the physiological or experimental buffer solution, often with MOPS as the pH buffer, as our main vehicle for primary fixatives composed with aldehydes, aldehyde-tannic-acid, or tannic acid alone, because we believe this offers the best hope for preserving native structure of various muscle structural states (but also effective with other cells and tissues). "Physiological" mostly implies proper ions: for extracellular salines (Na+ at ~100-145 mM; K+ absent or below 2 mM; Mg++ and Ca++ at 1-4 mM); or intracellular rest-state salines (EGTA and no inadvertent Ca++; Mg at ~5-7 mM; K+ (or Na+) up to 150 mM if desired; chloride, propionate, acetate, or methane-sulfonate as anion).
So for intact frog leg muscle (or insect flight muscle!) we used frog Ringer solution, for mammalian muscle we used mammalian Ringer, with either phosphate or MOPS buffer, adjusting pH with a "dirty" (fixative only) pH electrode to discover the amount of alkali or acid needed to restore original pH after adding various fixative at various concentrations. The presence of up to 3 mM calcium and up to 10 mm magnesium, as dictated by various buffers, was essential in Ringer recipes, and presented no problem. We were happy because the fiber x-ray diffraction pattern and the EM itself showed these fixatives to give good preservation of various distinct pre-fixation native structural states, and i think this was not so for our brief long-ago trials of phosphate or s-collidine buffer. We usually ignored adjustments to osmolarity, but if tried, was always done by adding the osmolytge to the formulation, sucrose top Ringer, 500,000 MW dextran to intracellular buffer.
For secondary fixation, buffer choice need not be physiological. For aldehyde-only or aldehyde-TA primary fixation, post-fixing in OsO4 has routinely employed a Maupin-Pollard good-for-actin buffer (0.1M phosphate, pH 6, with 10 mM MgCl2, used ice cold).
Most of our work has been with chemically demembranated ("skinned"; by detergent-glycerol) muscle of rabbit, frog or insect, so we chose various composition of intracellular buffer to keep the organelles happy and in the desired functional state of the muscle machinery (rigor, Mg-ATP relaxed, Mg-ATP-Ca activating, or modified with nucleotide analogs (like AMPPNP) phosphate analogs (like vanadate). We have for 30 years used nothing but 20 mM MOPS buffer (relatively inexpensive, and good for pH 6.5-7.5; we prefer 6.8 for insect) for both the physiological and fixative versions of our solutions, typically with 5 mM each of additives like MgCl2, EGTA, Ca-EGTA, ATP, Na azide (stops microbial growth, arrest ATPase consumption by mitochondria). For years we strictly used potassium salts in preference to sodium to honor the cell's intracellular habit, but lately have slipped away from this dogma without bad results, drifting to more frequent use of sodium (as hydroxide to adjust pH of EGTA, ATP, MOPS, etc. Sometimes we use salt to approximate intracellular ionic strength, typically using KCl (now NaCl) at 100 or 150 ml, but we often use no extra salt at all, because its presence or absence has so far made little or no difference to the fiber x-ray diffraction patterns which are our gold standard for native and preserved structure.
We have always steered clear of Tris because glutaraldehyde reacts with it, changing pH (?? and what else?), and because Tris-buffered pH alters with changing temperature.
0.2% tannic acid alone in the physiological (intracellular type) buffer works well on skinned cells to fix all but soluble components. It is blocked or complexed so it becomes unavailable for fixation when Triton X-100 or PVP are included in the same solution. TA-fix should be followed after rinse-out by uranyl acetate (typically in DI water) or OsO4 (in Maupin-Pollard good-for-actin buffer) as a secondary fixative.
-mike reedy-
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************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
This discussion takes me back to my doctoral research, in which I was concerned about the morphology of dissociated chick retina cells as they reassociated to form tissue-like aggregates. (Develop. Biol. 23:36-61, 1970(!)).
As part of that study, we fixed suspensions of cells in various concentrations of phosphate buffer, and measured the cell size distribution in the population with a Coulter Counter. We then chose to fix in 0.08 molar phosphate with 2.5% glutaraldehyde since that distribution matched the result with living cells.
It is clear that one can't just calculate osmotic strength for these systems, since the fixative alters the membrane behavior in a non- linear way during the fixation process.
Joel
-- Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 0486 http://astro.temple.edu/~jbs
==============================Original Headers============================== 9, 27 -- From jbs-at-temple.edu Wed Oct 11 11:03:44 2006 9, 27 -- Received: from po-smtp1.temple.edu (po-smtp1.temple.edu [155.247.166.195]) 9, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BG3gbR002357 9, 27 -- for {microscopy-at-microscopy.com} ; Wed, 11 Oct 2006 11:03:43 -0500 9, 27 -- Received: from [155.247.98.40] (jbs.bio.temple.edu [155.247.98.40]) 9, 27 -- by po-smtp1.temple.edu (MOS 3.8.2-GA) 9, 27 -- with ESMTP id FXB69399 (AUTH jbs); 9, 27 -- Wed, 11 Oct 2006 12:03:40 -0400 (EDT) 9, 27 -- From: "Joel Sheffield" {jbs-at-temple.edu} 9, 27 -- To: microscopy-at-microscopy.com 9, 27 -- Date: Wed, 11 Oct 2006 12:01:47 -0400 9, 27 -- MIME-Version: 1.0 9, 27 -- Subject: Re: [Microscopy] Re: debating about buffers for TEM fixation 9, 27 -- Reply-to: jbs-at-temple.edu 9, 27 -- Message-ID: {452CDD2B.17834.904B15E2-at-jbs.temple.edu} 9, 27 -- Priority: normal 9, 27 -- In-reply-to: {200610111534.k9BFYeEf023954-at-ns.microscopy.com} 9, 27 -- X-mailer: Pegasus Mail for Windows (4.30 public beta 1) 9, 27 -- Content-type: text/plain; charset=US-ASCII 9, 27 -- Content-transfer-encoding: 7BIT 9, 27 -- Content-description: Mail message body 9, 27 -- X-Junkmail-Status: score=10/50, host=po-smtp1.temple.edu 9, 27 -- X-Junkmail-SD-Raw: score=unknown, 9, 27 -- refid=str=0001.0A09020A.452D1597.00CD,ss=1,fgs=0, 9, 27 -- ip=155.247.98.40, 9, 27 -- so=2006-09-22 03:48:54, 9, 27 -- dmn=5.1.5/2006-01-31 ==============================End of - Headers==============================
Dear Winnie, The biggest problem with wood and, to a lesser extent charcoal, is that it is very porous, has water in its structure and will out-gas a lot of water vapour. Fresh concrete has the same problem. I have had to cut my wood samples down to less than 10 mm. cube in order to get the SEM to pump down to high vacuum. If you are looking at the wood samples in a variable-pressure SEM it should not be such a big problem. Most modern SEMs, unless they are very high resolution, will look at a 100 mm square sample, it is the pumping capability that might be a problem. Regards,
Mary Mager Electron Microscopist Materials Eng. UBC #419 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA Phone: 604-822-5648 Fax: 604-822-3619 email: mager-at-interchange.ubc.ca -----Original Message----- X-from: winnie.wino-at-gmail.com [mailto:winnie.wino-at-gmail.com] Sent: Wednesday, October 11, 2006 6:20 AM To: mager-at-interchange.ubc.ca
This Question was submitted to Ask-A-Microscopist by (winnie.wino-at-gmail.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, October 11, 2006 at 04:28:32 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both winnie.wino-at-gmail.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: winnie.wino-at-gmail.com Name: Winnie
Organization: NUS
Education: Undergraduate College
Location: Singapore
Title: Sample preparation
Question: Dear Microscopist, I have a few specimens (Charcoal and virgin wood) of which the sizes are 100x100x50(mm) and 30x30x100(mm). What should I prepare for those specimens before bringing them to the Lab for SEM scanning? Can the SEM machine's chamber accomodate such specimens? Should I reduce the size? Thank you and Best Regards. Winnie
I have used serum free media for many cell culture fixations. I think it works fine. After fix I go into CaCo so that I do not have any interactions with my TC media and any post-fix steps. David
On Oct 11, 2006, at 8:25 AM, drk-at-shcc.org wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Fellow Microscopists, } } Buffers have been on my mind lately too. In this laboratory, we have } switched our buffer system over to Dulbecco's DMEM cell culture } medium. In } large part this is a matter of convenience. We do quite a few } immunocytochemistry experiments, labeling culture cell membranes } with a } variety of antibodies, and we dilute these antibodies and subsequent } colloidal gold conjugates with DMEM. The cells remain viable during } incubations in antibody and secondary conjugates. It seemed } reasonable to } us to continue with DMEM during fixation, and we therefore dilute our } cacodylate buffered 3%glut/3%paraformaldehyde stock solution with } DMEM and } also our OsO4 in DMEM. We have been pleased with the quality of the } fixation } and now we also use DMEM for buffering fixatives of tissues with } equally } pleasing results. But I have to admit to a nagging feeling that I am } breaking some kind of microscopy law and would appreciate comments. } } Thanks, } } Doug } } Douglas R. Keene } Assistant Investigator } Micro-Imaging Center } Shriners Hospital for Children } 3102 S.W. Sam Jackson Park Road } Portland, Oregon 97239 } 503-221-3434 } drk-at-shcc.org } } -----Original Message----- } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: Wednesday, October 11, 2006 1:15 AM } To: drk-at-shcc.org } Subject: [Microscopy] debating about buffers for TEM fixation } } } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Wealthy colleagues (yes: knowledge is a wealth!), } } Perhaps we could exchange our views about the optimal } buffers to use during fixations for TEM. } Cacodylate is a traditional buffer, but it is not } really healthy and perhaps it is time to update our } knowledge. I have read that PIPES offered the lowest } extraction but on the downside a less effecient } buffering. Phosphate buffer does not allow the use of } Ca and Mg, which could help stabilize some structures. } HEPES buffer...well I don't know the downside, perhaps } it offers only advantages ;-)but it is used pretty } scarcely by the electron microscopists. } Some other parameters can be taken into account: the } storage, bench (or fridge) life, } morphology {} immunology, plants {} animals cells and so } on. } } Lets talk about salts! } } Personally I use Cacodylate for classical fixation of } cells for morphology because it has also been used in } the laboratory but I am eager to change. There is } certainly some room for improvements in the morphology } (especially of the membranes). HEPES seems to be a } good candidate. } } Stephane } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Wed Oct 11 03:09:36 2006 } 6, 18 -- Received: from web37409.mail.mud.yahoo.com } (web37409.mail.mud.yahoo.com [209.191.91.141]) } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } k9B89aiA016895 } 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 11 Oct 2006 03:09:36 } -0500 } 6, 18 -- Received: (qmail 28241 invoked by uid 60001); 11 Oct 2006 } 08:09:36 } -0000 } 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 6, 18 -- s=s1024; d=yahoo.com; } 6, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content- } Type:Content } -Transfer-Encoding; } 6, 18 -- } b=ChPAJbr1a2yJNpAafId35uxgkFOweiyNadUClf3kn/3vCzkXkmVSq83kMSU39wM/ } SA9OE32fVd } Mpo7cVyn14pAUg5FAqfmoj/NjpDIBgezrja3Sg7yKHXl4a7DvEOWxRqWDD+BDfBGD/ } GIFHMAZw8Q } 9GrzBsnlVdSy1r3b2snjk= ; } 6, 18 -- Message-ID: } {20061011080936.28239.qmail-at-web37409.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by } web37409.mail.mud.yahoo.com via } HTTP; Wed, 11 Oct 2006 01:09:36 PDT } 6, 18 -- Date: Wed, 11 Oct 2006 01:09:36 -0700 (PDT) } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 6, 18 -- Subject: debating about buffers for TEM fixation } 6, 18 -- To: microscopy-at-microscopy.com } 6, 18 -- MIME-Version: 1.0 } 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 6, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== } } } ==============================Original } Headers============================== } 17, 21 -- From drk-at-shcc.org Wed Oct 11 10:19:56 2006 } 17, 21 -- Received: from SUNNIE.shcc.org (mail.shcc.org } [64.213.211.202]) } 17, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k9BFJtsx013068 } 17, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 } 10:19:55 -0500 } 17, 21 -- Received: from DRK2 ([64.213.211.62]) by shcc.org (PMDF } V6.3 #31278) } 17, 21 -- with ESMTP id {0J6Z00E0V8V2X5-at-shcc.org} for } Microscopy-at-Microscopy.Com; Wed, } 17, 21 -- 11 Oct 2006 08:11:26 -0700 (PDT) } 17, 21 -- Date: Wed, 11 Oct 2006 08:19:49 -0700 } 17, 21 -- From: Doug Keene {drk-at-shcc.org} } 17, 21 -- Subject: RE: [Microscopy] debating about buffers for TEM } fixation } 17, 21 -- In-reply-to: {200610110814.k9B8EqC0024591-at-ns.microscopy.com} } 17, 21 -- To: Microscopy-at-Microscopy.Com } 17, 21 -- Reply-to: drk-at-shcc.org } 17, 21 -- Message-id: {0J6Z00E0W8V2X5-at-shcc.org} } 17, 21 -- Organization: Shriners Hospitals for Children } 17, 21 -- MIME-version: 1.0 } 17, 21 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 } 17, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 } 17, 21 -- Content-type: text/plain; charset=us-ascii } 17, 21 -- Content-transfer-encoding: 7bit } 17, 21 -- Thread-Index: AcbtDChpSOQbSAYeQd2UYXPEktNGrQAOl+9g } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-arizona.edu Wed Oct 11 11:47:04 2006 5, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BGl47S024189 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Oct 2006 11:47:04 -0500 5, 22 -- Received: from localhost (legolas.email.arizona.edu [10.0.0.224]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 05AB411299C2 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Oct 2006 09:46:48 -0700 (MST) 5, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id C9F4E1129080 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Oct 2006 09:40:16 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 22 -- In-Reply-To: {200610111525.k9BFPInw018425-at-ns.microscopy.com} 5, 22 -- References: {200610111525.k9BFPInw018425-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {699624AD-10B7-4A90-BC11-A49FBBD99E1D-at-arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-arizona.edu} 5, 22 -- Subject: Re: [Microscopy] RE: debating about buffers for TEM fixation 5, 22 -- Date: Wed, 11 Oct 2006 09:40:14 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.752.2) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Greetings, Mike Reedy mentioned that he uses MOPS for a buffer at pH 7 for fixations. I have always used PIPES. Has anyone ever compared the performance of these two Good buffers around neutral pH?? Are there reasons to choose one vs the other?
On Oct 11, 2006, at 10:04 AM, baskin-at-bio.umass.edu wrote:
} Mike Reedy mentioned that he uses MOPS for a buffer at pH 7 } for fixations. I have always used PIPES. Has anyone ever compared the } performance of these two Good buffers around neutral pH?? Are there } reasons to choose one vs the other? } Dear Tobias, The ability of a buffer to maintain pH depends on how many groups there are on the molecule that can take up or release H+ and the pK of each of these groups. For most buffers there are one or a few of these groups that have different pKs; e.g. PO4--- with pKs of ~12, ~7, and ~3. I don't know offhand what pKs are available for either MOPS or PIPES, but the closer a pK is to the pH you want, the better the buffer will be at maintaining that pH. The reason for this is easily seen from the mass action law, which can be written
K = [A][H]/[AH]
from which it can be seen that for [H] = K, [A] = [AH], and if [H] = K, then pH = pK. When the protonated and unprotonated species are at equal concentration, the addition of a small amount of acid or base will change the [A]/[AH] ratio less than if they are at unequal concentration. Therefore, the performance of either Good buffer will equal that of the other one if the desired pH is the same amount away from either pK. Another reason to choose one over another is if one of them affects the process you are trying to perform. It may be that one will react with your fixative--which I do not think is the case for MOPS or PIPES and either glut or OsO4. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 6, 22 -- From tivol-at-caltech.edu Wed Oct 11 14:52:43 2006 6, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BJqh5u016670 6, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 14:52:43 -0500 6, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 6, 22 -- by water-ox-postvirus (Postfix) with ESMTP id CEFFF2EF2E 6, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 12:52:42 -0700 (PDT) 6, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 6, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 64B9C109AED 6, 22 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 12:52:41 -0700 (PDT) 6, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 6, 22 -- In-Reply-To: {200610111704.k9BH4Ypv002703-at-ns.microscopy.com} 6, 22 -- References: {200610111704.k9BH4Ypv002703-at-ns.microscopy.com} 6, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 6, 22 -- Message-Id: {4cb53a98ada174cee4b63e632eab0ca7-at-caltech.edu} 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 6, 22 -- Subject: Re: [Microscopy] MOPS vs PIPES 6, 22 -- Date: Wed, 11 Oct 2006 12:56:56 -0700 6, 22 -- To: microscopy-at-msa.microscopy.com 6, 22 -- X-Mailer: Apple Mail (2.624) 6, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
What is your experience with the film camera? How many successful film cassettes do you average between camera jams? Please quote for each TEM model (100CX, 2000FX, 3100, etc).
Thanks, Roseann
Roseann Csencsits, PhD. Donner TEM Facility Manager Lawrence Berkeley Lab 510-486-4548
==============================Original Headers============================== 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} 7, 21 -- Subject: JEOL camera question 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 7, 21 -- To: Microscopy-at-Microscopy.Com 7, 21 -- X-Mailer: Apple Mail (2.752.3) 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 7, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
I have been reading this thread with interest in the hope of learning something new. The theory of buffers as explained by Bill Tivol, doesn't seem to be useful when applied to electron microscopy. The reason is that we judge the effects of different buffers on their effect on morphology (as Mike Reedy subtly puts it in his message), not on the final pH of the specimens.
Judged on buffering capacity alone, sodium cacodylate would fail miserably as a buffer. The Good buffers (PIPES, HEPES etc) appear to be excellent buffers for keeping solutions at a stable pH. However, even if PIPES or HEPES is used to buffer aldehyde, when the solution encounters biological material there is a sudden drop in pH that is not affected at all by the buffer in use. Anyone who has fixed cell cultures in the presence of medium containing pH indicator can attest to the rapid color change that occurs.
It is worth noting that the pH to which buffers are adjusted, and the pH at which they are most effective, is not always 7.2 or 7.4. For example, the use of PIPES at a pH higher than 6.8 will greatly reduce its buffering capacity.
I liked the idea of comparing the sizes of fixed cells with living cells to determine which concentration of buffer is "best" to use. There was no mention of what the intracellular morphology looked like and if it was acceptable. I am sure that the best concentration will differ between cell types. The great disadvantage of this method for determining "correct' buffer concentrations is that most of our work involves tissues composed of many different cell types that are all attached to each other.
Another issue touched on by Mike Reedy is the state of aldehyde-fixed biological material. Is is really osmotically inactive as assumed (e.g. "...buffer choice need not be physiological.")? Could aqueous solutions of osmium tetroxide be used in post-fixation steps? Are aldehyde-fixed membranes osmotically inactive?
Our final evaluation of buffers will depend on what we are looking at, or looking for, in our specimens. Sometimes we need to extract lots of cytoplasmic components to give high-contrast images. Other times we need to retain as many proteins as possible so that we can perform immunocytochemical labeling. Other important considerations for immunolabeling are the need to retain all antigens at their "natural" site, yet enable complete antibody accessibility to antigens.
For the record, our routine fixation protocols involve glutaraldehyde and sodium cacodylate for morphology, and double-strength aldehyde (formaldehyde or glutaraldehyde, or mixtures of both) in double strength HEPES buffer for immunocytochemistry. The double-strength fixative is added to culture medium, ringer or other physiological medium. There routine fixatives work well for us, but we do not work with sharks, molluscs or plants.
Any EM laboratory involved in many projects may find that one fixation protocol is insufficient for all users. Instead, different protocols involving different buffers gradually evolve in the lab to cope with each specimen as needed. What we need is a good understanding of how buffers and cross-linking chemicals affect the specimens we apply them to.
Regards,
Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 (213) 273 8026 pwebster-at-hei.org
The ability of a buffer to maintain pH depends on how many groups there are on the molecule that can take up or release H+ and the pK of each of these groups. For most buffers there are one or a few of these groups that have different pKs; e.g. PO4--- with pKs of ~12, ~7, and ~3. I don't know offhand what pKs are available for either MOPS or PIPES, but the closer a pK is to the pH you want, the better the buffer will be at maintaining that pH. The reason for this is easily seen from the mass action law, which can be written
K = [A][H]/[AH]
from which it can be seen that for [H] = K, [A] = [AH], and if [H] = K, then pH = pK. When the protonated and unprotonated species are at equal concentration, the addition of a small amount of acid or base will change the [A]/[AH] ratio less than if they are at unequal concentration. Therefore, the performance of either Good buffer will equal that of the other one if the desired pH is the same amount away from either pK. Another reason to choose one over another is if one of them affects the process you are trying to perform. It may be that one will react with your fixative--which I do not think is the case for MOPS or PIPES and either glut or OsO4. Yours, Bill Tivol, PhD
For 40 years we have used the physiological or experimental buffer solution, often with MOPS as the pH buffer, as our main vehicle for primary fixatives composed with aldehydes, aldehyde-tannic-acid, or tannic acid alone, because we believe this offers the best hope for preserving native structure of various muscle structural states (but also effective with other cells and tissues). "Physiological" mostly implies proper ions: for extracellular salines (Na+ at ~100-145 mM; K+ absent or below 2 mM; Mg++ and Ca++ at 1-4 mM); or intracellular rest-state salines (EGTA and no inadvertent Ca++; Mg at ~5-7 mM; K+ (or Na+) up to 150 mM if desired; chloride, propionate, acetate, or methane-sulfonate as anion).
So for intact frog leg muscle (or insect flight muscle!) we used frog Ringer solution, for mammalian muscle we used mammalian Ringer, with either phosphate or MOPS buffer, adjusting pH with a "dirty" (fixative only) pH electrode to discover the amount of alkali or acid needed to restore original pH after adding various fixative at various concentrations. The presence of up to 3 mM calcium and up to 10 mm magnesium, as dictated by various buffers, was essential in Ringer recipes, and presented no problem. We were happy because the fiber x-ray diffraction pattern and the EM itself showed these fixatives to give good preservation of various distinct pre-fixation native structural states, and i think this was not so for our brief long-ago trials of phosphate or s-collidine buffer. We usually ignored adjustments to osmolarity, but if tried, was always done by adding the osmolytge to the formulation, sucrose top Ringer, 500,000 MW dextran to intracellular buffer.
For secondary fixation, buffer choice need not be physiological. For aldehyde-only or aldehyde-TA primary fixation, post-fixing in OsO4 has routinely employed a Maupin-Pollard good-for-actin buffer (0.1M phosphate, pH 6, with 10 mM MgCl2, used ice cold).
Most of our work has been with chemically demembranated ("skinned"; by detergent-glycerol) muscle of rabbit, frog or insect, so we chose various composition of intracellular buffer to keep the organelles happy and in the desired functional state of the muscle machinery (rigor, Mg-ATP relaxed, Mg-ATP-Ca activating, or modified with nucleotide analogs (like AMPPNP) phosphate analogs (like vanadate). We have for 30 years used nothing but 20 mM MOPS buffer (relatively inexpensive, and good for pH 6.5-7.5; we prefer 6.8 for insect) for both the physiological and fixative versions of our solutions, typically with 5 mM each of additives like MgCl2, EGTA, Ca-EGTA, ATP, Na azide (stops microbial growth, arrest ATPase consumption by mitochondria). For years we strictly used potassium salts in preference to sodium to honor the cell's intracellular habit, but lately have slipped away from this dogma without bad results, drifting to more frequent use of sodium (as hydroxide to adjust pH of EGTA, ATP, MOPS, etc. Sometimes we use salt to approximate intracellular ionic strength, typically using KCl (now NaCl) at 100 or 150 ml, but we often use no extra salt at all, because its presence or absence has so far made little or no difference to the fiber x-ray diffraction patterns which are our gold standard for native and preserved structure.
We have always steered clear of Tris because glutaraldehyde reacts with it, changing pH (?? and what else?), and because Tris-buffered pH alters with changing temperature.
0.2% tannic acid alone in the physiological (intracellular type) buffer works well on skinned cells to fix all but soluble components. It is blocked or complexed so it becomes unavailable for fixation when Triton X-100 or PVP are included in the same solution. TA-fix should be followed after rinse-out by uranyl acetate (typically in DI water) or OsO4 (in Maupin-Pollard good-for-actin buffer) as a secondary fixative.
-mike reedy-
==============================Original Headers============================== 31, 18 -- From PWebster-at-hei.org Wed Oct 11 16:18:24 2006 31, 18 -- Received: from hi0sml1.hei.org (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) 31, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BLIOUn006955 31, 18 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 16:18:24 -0500 31, 18 -- Received: from 10.10.42.113 ([10.10.42.113]) by hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server HTTP-DAV ; 31, 18 -- Wed, 11 Oct 2006 21:18:23 +0000 31, 18 -- User-Agent: Microsoft-Entourage/11.2.5.060620 31, 18 -- Date: Wed, 11 Oct 2006 14:18:23 -0700 31, 18 -- Subject: Re: debating about buffers for TEM fixation 31, 18 -- From: "Webster, Paul" {PWebster-at-hei.org} 31, 18 -- To: {microscopy-at-msa.microscopy.com} 31, 18 -- Message-ID: {C152ADAF.C5ED%PWebster-at-hei.org} 31, 18 -- Thread-Topic: debating about buffers for TEM fixation 31, 18 -- Thread-Index: AcbteshdByXfOlluEdua3wANk7Zh7g== 31, 18 -- Mime-version: 1.0 31, 18 -- Content-type: text/plain; 31, 18 -- charset="US-ASCII" 31, 18 -- Content-transfer-encoding: 7bit ==============================End of - Headers==============================
Roseann; My experience is that it is more a function of care and attention to detail of loading film into the cassettes than anything else.
John Mardinly Intel Corporation
Disclaimer: This is the opinion of this individual, not the opinion of Intel Corporation.
-----Original Message----- X-from: rcsencsits-at-lbl.gov [mailto:rcsencsits-at-lbl.gov] Sent: Wednesday, October 11, 2006 2:07 PM To: Mardinly, John
Dear JEOL film camera users:
What is your experience with the film camera? How many successful film cassettes do you average between camera jams? Please quote for each TEM model (100CX, 2000FX, 3100, etc).
Thanks, Roseann
Roseann Csencsits, PhD. Donner TEM Facility Manager Lawrence Berkeley Lab 510-486-4548
==============================Original Headers============================== 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} 7, 21 -- Subject: JEOL camera question 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 7, 21 -- To: Microscopy-at-Microscopy.Com 7, 21 -- X-Mailer: Apple Mail (2.752.3) 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 7, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
==============================Original Headers============================== 16, 31 -- From john.mardinly-at-intel.com Wed Oct 11 19:48:10 2006 16, 31 -- Received: from mga03.intel.com (mga03.intel.com [143.182.124.21]) 16, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9C0m9Ma021539 16, 31 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 19:48:10 -0500 16, 31 -- Received: from azsmga001.ch.intel.com ([10.2.17.19]) 16, 31 -- by mga03.intel.com with ESMTP; 11 Oct 2006 17:48:07 -0700 16, 31 -- Received: from scsmsx332.sc.intel.com (HELO scsmsx332.amr.corp.intel.com) ([10.3.90.6]) 16, 31 -- by azsmga001.ch.intel.com with ESMTP; 11 Oct 2006 17:48:07 -0700 16, 31 -- X-ExtLoop1: 1 16, 31 -- X-IronPort-AV: i="4.09,297,1157353200"; 16, 31 -- d="scan'208"; a="129889456:sNHT24905965" 16, 31 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.211); 16, 31 -- Wed, 11 Oct 2006 17:48:06 -0700 16, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 16, 31 -- Content-class: urn:content-classes:message 16, 31 -- MIME-Version: 1.0 16, 31 -- Content-Type: text/plain; 16, 31 -- charset="us-ascii" 16, 31 -- Subject: RE: [Microscopy] JEOL camera question 16, 31 -- Date: Wed, 11 Oct 2006 17:48:05 -0700 16, 31 -- Message-ID: {1DF4C4D62339DB4C9C98DF04213995B60170A5A2-at-scsmsx413.amr.corp.intel.com} 16, 31 -- X-MS-Has-Attach: 16, 31 -- X-MS-TNEF-Correlator: 16, 31 -- Thread-Topic: [Microscopy] JEOL camera question 16, 31 -- Thread-Index: AcbteTQlou9AYV+TSiKWxqGREJ+P3AAHfgrA 16, 31 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 16, 31 -- To: {rcsencsits-at-lbl.gov} 16, 31 -- Cc: {Microscopy-at-msa.microscopy.com} 16, 31 -- X-OriginalArrivalTime: 12 Oct 2006 00:48:06.0868 (UTC) FILETIME=[14EF5940:01C6ED98] 16, 31 -- Content-Transfer-Encoding: 8bit 16, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9C0m9Ma021539 ==============================End of - Headers==============================
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Email: kychung2-at-wisc.edu Name: Ka Young Chung
Organization: University of Wisconsin-Madison
Title-Subject: [Filtered] Fixation problem
Question: I am working with cardiac myocytes. To perform immunofluorescence, I have permeabilized cells with saponin without fixation, and could get nice signal. The signal was blocked when I used blocking peptide (original immunogen for primary antibody). To look at the localization pattern more in detail, I want to do electron microscopy. In this case, I need to fix the cells. To check whether fixation affects antibody interaction, I fixed cells with 4% paraformaldehyde and 0.1% glutaraldehyde, permeabilized cells with saponin, and then performed immunofluorecence. I saw a permeabilization problem with this experiment so that I tried 0.1% triton instead of saponin. Still, I am not getting the same staining pattern that I got from unfixed cells. How can I solve this problem? Can I just try EM?
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Question: I am searching for info.relating to an antique bausch&lomb metallograph. If anyone has related info. it would be so very much appreciated. I can email photos of the metallogragh to anyone who might have the knowledge to enlighten me about this. It is in good working condition,with many accesories ,assorted lenses and interchangable cameras. It is a large free standind, two pedestal, piece of equiptment, with camera like devices at each end. Stamp into it is the following info.
BAUSCH & LOMB OPTICAL CO. ROCHESTER, NY 01348
one div. =0.002mm Serial No.-6097 US Pat. 1798634 2074106
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Email: mdelann1-at-jhmi.edu Name: Michael Delannoy
Organization: Assistant Director Imaging Facility JHSM
Title-Subject: [Filtered] chillers
Question: Hello I need a few companies who would sell used/ refurbished water chillers/recirculators. I need a supply for my LEO 1530 FESEM Does anyone know the heat capaciy required? Does anyone know the Neslabs website or contact info? Thank You M Delannoy 410 955-1365
We have a Hitachi H-600 TEM, 1983. For the past year we've been operating with a faulty CRT data display unit. Each morning the display would come on for about ten minutes, then it would become unstable and disappear for the rest of the day. Finally it appears to have died completely which makes operating the TEM tricky. Our in-house electrician has had several attempts at fixing the problem but without success. The microscope itself is completely functional so operations such as column alignment, resetting number of unexposed negatives, etc are still possible but have to be done blind. As you can imagine this is frustrating and there is great potential for a mix-up with regard to matching specimens to negative numbers.
I have some questions for you...
Are these CRT data display units unique for this Hitachi model or are units from other models compatible with the H-600? Can CRT units from other electrical equipment be sourced, eg radiology machines? Can new CRT units be bought today that would be compatible? Does anyone have an old working unit that they would be willing to part with? Has anyone had similar experiences with CRT units? Does our unit sound fixable or are we wasting our time?
Thanks,
John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia Australia
john.brealey-at-imvs.sa.gov.au
(08) 8222 6612
==============================Original Headers============================== 3, 33 -- From john.brealey-at-imvs.sa.gov.au Wed Oct 11 21:02:16 2006 3, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 3, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9C22FMx032484 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 11 Oct 2006 21:02:16 -0500 3, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 3, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k9C22FJZ024600 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 11:32:15 +0930 (CST)' 3, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 3, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k9C22FmJ024594 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 11:32:15 +0930 (CST)' 3, 33 -- Received: from localhost (scalix1.imvs.sa.gov.au [10.20.98.38]) 3, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id F350934F22 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 11:32:14 +0930 (CST) 3, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 3, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 3, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.38]) (amavisd-new, port 10024) 3, 33 -- with LMTP id Yz015DgQqpt4 for {Microscopy-at-MSA.Microscopy.Com} ; 3, 33 -- Thu, 12 Oct 2006 11:32:04 +0930 (CST) 3, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 3, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 89C8F34F19 3, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 11:31:28 +0930 (CST) 3, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 3, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 3, 33 -- Subject: Hitachi H-600 CRT Problem 3, 33 -- Date: Thu, 12 Oct 2006 11:31:27 +0930 3, 33 -- Message-ID: {000001c6eda2$549789a0$c88a140a-at-iqe36042} 3, 33 -- MIME-Version: 1.0 3, 33 -- Content-Type: text/plain; 3, 33 -- charset="us-ascii" 3, 33 -- Content-Transfer-Encoding: 7bit 3, 33 -- X-Mailer: Microsoft Office Outlook 11 3, 33 -- Thread-Index: AcbtokEn2wZh7yMrRRGmgQ4kZRU8jA== 3, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 ==============================End of - Headers==============================
I run several JEOL TEMs with film cameras (200CX, 2000FX, 2010, 4000) with a large contingent of new users every year. We do get the occasional jam due to misloaded film cassettes by one of the new users. Apart from that we rarely experience camera jams and I would estimate that at the height of our film use the number of films between jams was well in the 10,000's. Film use has declined over the past few years as we have some digital cameras. As far as I can see the camera mechanism has not changed since our 100B so there is no reason to have a different failure rate on any instrument you listed.
If you are experiencing problems with camera jams then check that your cassettes and boxes are not damaged. Check that the films are firmly held in all four corners when they are loaded, very occasionally we get undersized film that will not fit properly. If these are OK then there may be a problem with your camera mechanism, possibly damaged when freeing off a jammed cassette. If you know how to remove the mechanism check that the leaf springs on the pushers are OK and that the tracks are not damaged.
Ron
-----Original Message----- X-from: rcsencsits-at-lbl.gov [mailto:rcsencsits-at-lbl.gov] Sent: 11 October 2006 22:15 To: ron.doole-at-materials.ox.ac.uk
Dear JEOL film camera users:
What is your experience with the film camera? How many successful film cassettes do you average between camera jams? Please quote for each TEM model (100CX, 2000FX, 3100, etc).
Thanks, Roseann
Roseann Csencsits, PhD. Donner TEM Facility Manager Lawrence Berkeley Lab 510-486-4548
==============================Original Headers============================== 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} 7, 21 -- Subject: JEOL camera question 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 7, 21 -- To: Microscopy-at-Microscopy.Com 7, 21 -- X-Mailer: Apple Mail (2.752.3) 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 7, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
==============================Original Headers============================== 20, 26 -- From ron.doole-at-materials.ox.ac.uk Thu Oct 12 02:35:25 2006 20, 26 -- Received: from relay5.mail.ox.ac.uk (relay5.mail.ox.ac.uk [163.1.2.163]) 20, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9C7ZO0k016145 20, 26 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 02:35:25 -0500 20, 26 -- Received: from smtp1.mail.ox.ac.uk ([129.67.1.207]) 20, 26 -- by relay5.mail.ox.ac.uk with esmtp (Exim 4.62) 20, 26 -- (envelope-from {ron.doole-at-materials.ox.ac.uk} ) 20, 26 -- id 1GXv6C-0000nq-Gx; Thu, 12 Oct 2006 08:35:24 +0100 20, 26 -- Received: from oums-doole.materials.ox.ac.uk ([129.67.85.58] helo=oumsdoole) 20, 26 -- by smtp1.mail.ox.ac.uk with esmtp (Exim 4.62) 20, 26 -- (envelope-from {ron.doole-at-materials.ox.ac.uk} ) 20, 26 -- id 1GXv6B-0004IX-6L; Thu, 12 Oct 2006 08:35:24 +0100 20, 26 -- From: "Ron Doole" {ron.doole-at-materials.ox.ac.uk} 20, 26 -- To: {rcsencsits-at-lbl.gov} 20, 26 -- Cc: "'Microscopy Society of America'" {Microscopy-at-MSA.Microscopy.Com} 20, 26 -- Subject: RE: [Microscopy] JEOL camera question 20, 26 -- Date: Thu, 12 Oct 2006 08:35:22 +0100 20, 26 -- Message-ID: {000101c6edd0$f9c31510$3a554381-at-oumsdoole} 20, 26 -- MIME-Version: 1.0 20, 26 -- Content-Type: text/plain; 20, 26 -- charset="us-ascii" 20, 26 -- Content-Transfer-Encoding: 7bit 20, 26 -- X-Mailer: Microsoft Office Outlook 11 20, 26 -- Thread-Index: AcbtelS7vQ0g8W45S6mwwOiH18xAxwAUmEuw 20, 26 -- In-Reply-To: {200610112115.k9BLF6PR006189-at-ns.microscopy.com} 20, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
It is not really surprising that you see a difference between the 2 protocols. Please keep in mind that without fixation and with mild permeabilization the antibodies probable have access only to a few compartments of the cell. Using other treatments perhaps allows the antibodies to have better access to the antigens. On the contrary, it is possible that fixation has hidden some antigenic sites. Or perhaps a combination of both effects! Also, be careful using glutaraldehyde in fluorescence since it gives background. I would suggest you try directly in immunoEM and don't lose too much time playing around with protocols because it can be that your antibody simply doesn't work in EM. If you get a signal in EM, you can always discuss its pattern and compare it with the pattern obtained in IF.
Good luck
Stephane
----- Original Message ---- X-from: "kychung2-at-wisc.edu" {kychung2-at-wisc.edu} To: nizets2-at-yahoo.com Sent: Thursday, October 12, 2006 3:43:12 AM
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 kychung2-at-wisc.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: kychung2-at-wisc.edu Name: Ka Young Chung
Organization: University of Wisconsin-Madison
Title-Subject: [Filtered] Fixation problem
Question: I am working with cardiac myocytes. To perform immunofluorescence, I have permeabilized cells with saponin without fixation, and could get nice signal. The signal was blocked when I used blocking peptide (original immunogen for primary antibody). To look at the localization pattern more in detail, I want to do electron microscopy. In this case, I need to fix the cells. To check whether fixation affects antibody interaction, I fixed cells with 4% paraformaldehyde and 0.1% glutaraldehyde, permeabilized cells with saponin, and then performed immunofluorecence. I saw a permeabilization problem with this experiment so that I tried 0.1% triton instead of saponin. Still, I am not getting the same staining pattern that I got from unfixed cells. How can I solve this problem? Can I just try EM?
An important topic. I'd like to step in with a reply to just the point about "Could aqueous solutions of osmium tetroxide be used in post-fixation steps?" Yes. Not only have I known folks who make their OsO4 in distilled water for TEM, I have studied cultured cells in high-resolution SEM using OsO4 in distilled water, not buffer, for post-fixation. Usually with 1% monomeric tannic acid (for plasma membrane preservation), but not always. Works fine. The colligative properties of the plasma membrane seem to be pretty much eliminate by fixation.
pH isn't the only confusing issue here -- osmotic properties are, too. 3% glut is about 300 mOsM itself, more if there are impurities and after adding buffer. Yet, this hypertonicity doesn't seem to cause osmotic problems.
Worse, something I've never seen discussed: what are the osmotic effects of different pHs? Changing the pH will change the flow of ions and water in and out of cells. If a fixative is present as the pH changes, then the changes in the membrane proteins affecting the osmotic properties will be fixed. So potentionally, a correct pH may in fact not be the physiological pH of the living cell, but the pH that maintains the correct osmotic pressure(s) during fixation.
Phil
} Dear all, } } I have been reading this thread with interest in the hope of learning } something new. The theory of buffers as explained by Bill Tivol, doesn't } seem to be useful when applied to electron microscopy. The reason is that we } judge the effects of different buffers on their effect on morphology (as } Mike Reedy subtly puts it in his message), not on the final pH of the } specimens. } } Judged on buffering capacity alone, sodium cacodylate would fail miserably } as a buffer. The Good buffers (PIPES, HEPES etc) appear to be excellent } buffers for keeping solutions at a stable pH. However, even if PIPES or } HEPES is used to buffer aldehyde, when the solution encounters biological } material there is a sudden drop in pH that is not affected at all by the } buffer in use. Anyone who has fixed cell cultures in the presence of medium } containing pH indicator can attest to the rapid color change that occurs. } } It is worth noting that the pH to which buffers are adjusted, and the pH at } which they are most effective, is not always 7.2 or 7.4. For example, the } use of PIPES at a pH higher than 6.8 will greatly reduce its buffering } capacity. } } I liked the idea of comparing the sizes of fixed cells with living cells to } determine which concentration of buffer is "best" to use. There was no } mention of what the intracellular morphology looked like and if it was } acceptable. I am sure that the best concentration will differ between cell } types. The great disadvantage of this method for determining "correct' } buffer concentrations is that most of our work involves tissues composed of } many different cell types that are all attached to each other. } } Another issue touched on by Mike Reedy is the state of aldehyde-fixed } biological material. Is is really osmotically inactive as assumed (e.g. } "...buffer choice need not be physiological.")? Could aqueous solutions of } osmium tetroxide be used in post-fixation steps? Are aldehyde-fixed } membranes osmotically inactive? } } Our final evaluation of buffers will depend on what we are looking at, or } looking for, in our specimens. Sometimes we need to extract lots of } cytoplasmic components to give high-contrast images. Other times we need to } retain as many proteins as possible so that we can perform } immunocytochemical labeling. Other important considerations for } immunolabeling are the need to retain all antigens at their "natural" site, } yet enable complete antibody accessibility to antigens. } } For the record, our routine fixation protocols involve glutaraldehyde and } sodium cacodylate for morphology, and double-strength aldehyde (formaldehyde } or glutaraldehyde, or mixtures of both) in double strength HEPES buffer for } immunocytochemistry. The double-strength fixative is added to culture } medium, ringer or other physiological medium. There routine fixatives work } well for us, but we do not work with sharks, molluscs or plants. } } Any EM laboratory involved in many projects may find that one fixation } protocol is insufficient for all users. Instead, different protocols } involving different buffers gradually evolve in the lab to cope with each } specimen as needed. What we need is a good understanding of how buffers and } cross-linking chemicals affect the specimens we apply them to. } } Regards, } } Paul Webster } } } Paul Webster, Ph.D } House Ear Institute } 2100 West Third Street } Los Angeles, CA 90057 } (213) 273 8026 } pwebster-at-hei.org } } } } } } } ____________________________________________________________________ } } The ability of a buffer to maintain pH depends on how many groups } there are on the molecule that can take up or release H+ and the pK of } each of these groups. For most buffers there are one or a few of these } groups that have different pKs; e.g. PO4--- with pKs of ~12, ~7, and } ~3. I don't know offhand what pKs are available for either MOPS or } PIPES, but the closer a pK is to the pH you want, the better the buffer } will be at maintaining that pH. The reason for this is easily seen } from the mass action law, which can be written } } K = [A][H]/[AH] } } from which it can be seen that for [H] = K, [A] = [AH], and if [H] = K, } then pH = pK. When the protonated and unprotonated species are at } equal concentration, the addition of a small amount of acid or base } will change the [A]/[AH] ratio less than if they are at unequal } concentration. Therefore, the performance of either Good buffer will } equal that of the other one if the desired pH is the same amount away } from either pK. Another reason to choose one over another is if one of } them affects the process you are trying to perform. It may be that one } will react with your fixative--which I do not think is the case for } MOPS or PIPES and either glut or OsO4. } Yours, } Bill Tivol, PhD } } ___________________________________________________________________ } } For 40 years we have used the physiological or experimental buffer } solution, often with MOPS as the pH buffer, as our main vehicle for } primary fixatives composed with aldehydes, aldehyde-tannic-acid, or } tannic acid alone, because we believe this offers the best hope for } preserving native structure of various muscle structural states (but } also effective with other cells and tissues). "Physiological" mostly } implies proper ions: for extracellular salines (Na+ at ~100-145 mM; } K+ absent or below 2 mM; Mg++ and Ca++ at 1-4 mM); or intracellular } rest-state salines (EGTA and no inadvertent Ca++; Mg at ~5-7 mM; K+ } (or Na+) up to 150 mM if desired; chloride, propionate, acetate, or } methane-sulfonate as anion). } } So for intact frog leg muscle (or insect flight muscle!) we used frog } Ringer solution, for mammalian muscle we used mammalian Ringer, with } either phosphate or MOPS buffer, adjusting pH with a "dirty" } (fixative only) pH electrode to discover the amount of alkali or acid } needed to restore original pH after adding various fixative at } various concentrations. The presence of up to 3 mM calcium and up to } 10 mm magnesium, as dictated by various buffers, was essential in } Ringer recipes, and presented no problem. We were happy because the } fiber x-ray diffraction pattern and the EM itself showed these } fixatives to give good preservation of various distinct pre-fixation } native structural states, and i think this was not so for our brief } long-ago trials of phosphate or s-collidine buffer. We usually } ignored adjustments to osmolarity, but if tried, was always done by } adding the osmolytge to the formulation, sucrose top Ringer, 500,000 } MW dextran to intracellular buffer. } } For secondary fixation, buffer choice need not be physiological. For } aldehyde-only or aldehyde-TA primary fixation, post-fixing in OsO4 } has routinely employed a Maupin-Pollard good-for-actin buffer (0.1M } phosphate, pH 6, with 10 mM MgCl2, used ice cold). } } Most of our work has been with chemically demembranated ("skinned"; } by detergent-glycerol) muscle of rabbit, frog or insect, so we chose } various composition of intracellular buffer to keep the organelles } happy and in the desired functional state of the muscle machinery } (rigor, Mg-ATP relaxed, Mg-ATP-Ca activating, or modified with } nucleotide analogs (like AMPPNP) phosphate analogs (like vanadate). } We have for 30 years used nothing but 20 mM MOPS buffer (relatively } inexpensive, and good for pH 6.5-7.5; we prefer 6.8 for insect) for } both the physiological and fixative versions of our solutions, } typically with 5 mM each of additives like MgCl2, EGTA, Ca-EGTA, ATP, } Na azide (stops microbial growth, arrest ATPase consumption by } mitochondria). For years we strictly used potassium salts in } preference to sodium to honor the cell's intracellular habit, but } lately have slipped away from this dogma without bad results, } drifting to more frequent use of sodium (as hydroxide to adjust pH of } EGTA, ATP, MOPS, etc. Sometimes we use salt to approximate } intracellular ionic strength, typically using KCl (now NaCl) at 100 } or 150 ml, but we often use no extra salt at all, because its } presence or absence has so far made little or no difference to the } fiber x-ray diffraction patterns which are our gold standard for } native and preserved structure. } } We have always steered clear of Tris because glutaraldehyde reacts } with it, changing pH (?? and what else?), and because Tris-buffered } pH alters with changing temperature. } } 0.2% tannic acid alone in the physiological (intracellular type) } buffer works well on skinned cells to fix all but soluble components. } It is blocked or complexed so it becomes unavailable for fixation } when Triton X-100 or PVP are included in the same solution. TA-fix } should be followed after rinse-out by uranyl acetate (typically in DI } water) or OsO4 (in Maupin-Pollard good-for-actin buffer) as a } secondary fixative. } } -mike reedy- } } } ==============================Original Headers============================== } 31, 18 -- From PWebster-at-hei.org Wed Oct 11 16:18:24 2006 } 31, 18 -- Received: from hi0sml1.hei.org } (House-Ear-Institute-1114861.cust-rtr.pacbell.net [71.136.31.242]) } 31, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k9BLIOUn006955 } 31, 18 -- for {microscopy-at-msa.microscopy.com} ; Wed, 11 Oct 2006 } 16:18:24 -0500 } 31, 18 -- Received: from 10.10.42.113 ([10.10.42.113]) by } hi0sml1.hei.org ([10.10.40.106]) with Microsoft Exchange Server } HTTP-DAV ; } 31, 18 -- Wed, 11 Oct 2006 21:18:23 +0000 } 31, 18 -- User-Agent: Microsoft-Entourage/11.2.5.060620 } 31, 18 -- Date: Wed, 11 Oct 2006 14:18:23 -0700 } 31, 18 -- Subject: Re: debating about buffers for TEM fixation } 31, 18 -- From: "Webster, Paul" {PWebster-at-hei.org} } 31, 18 -- To: {microscopy-at-msa.microscopy.com} } 31, 18 -- Message-ID: {C152ADAF.C5ED%PWebster-at-hei.org} } 31, 18 -- Thread-Topic: debating about buffers for TEM fixation } 31, 18 -- Thread-Index: AcbteshdByXfOlluEdua3wANk7Zh7g== } 31, 18 -- Mime-version: 1.0 } 31, 18 -- Content-type: text/plain; } 31, 18 -- charset="US-ASCII" } 31, 18 -- Content-transfer-encoding: 7bit } ==============================End of - Headers==============================
-- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 6, 22 -- From oshel1pe-at-cmich.edu Thu Oct 12 07:19:22 2006 6, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CCJM6W009892 6, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 07:19:22 -0500 6, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 6, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k9CCoWNA010683 6, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 08:50:41 -0400 6, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 6, 22 -- Thu, 12 Oct 2006 08:19:12 -0400 6, 22 -- Mime-Version: 1.0 6, 22 -- Message-Id: {f06230900c153e0a29915-at-[141.209.160.249]} 6, 22 -- In-Reply-To: {200610112122.k9BLM7xQ014251-at-ns.microscopy.com} 6, 22 -- References: {200610112122.k9BLM7xQ014251-at-ns.microscopy.com} 6, 22 -- Date: Thu, 12 Oct 2006 08:19:10 -0400 6, 22 -- To: Microscopy-at-microscopy.com 6, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 6, 22 -- Subject: [Microscopy] Re: debating about buffers for TEM fixation 6, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 22 -- X-OriginalArrivalTime: 12 Oct 2006 12:19:12.0354 (UTC) FILETIME=[A04A6420:01C6EDF8] 6, 22 -- X-CanItPRO-Stream: default 6, 22 -- X-Spam-Score: -3.9 () L_EXCH_MF,TW_YT 6, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
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Email: U.J.Potter Name: Ursula Potter
Organization: University of Bath
Title-Subject: [Filtered] Neg carriers jamming in JEOL EM
Question: Dear All,
I managed to erase Rosannes message about negative cassettes jamming in JEOL TEM film cameras before I had replied - so hope she sees it here.
We have had this problem and found it was due to warped cassettes. We scratched a very fine number on each cassette and then found it was the same cassette or couple of cassettes causing the problem every time. Since numbering the cassettes and weeding out the troublesome ones we have had very few jamming problems.
Hope this helps.
Ursula ------------------------- CEOS University of Bath UK
I cannot recall having any camera jams with our JEOL 2011. Our older Philips EM420 has periodic jams- usually due to bent film plates or poorly loaded film.
Cheers Roger A. Ristau, PhD Electron Microscopy Specialist Institute of Materials Science 97 North Eagleville Road University of Connecticut Storrs, CT 06269
} } Dear JEOL film camera users: } } What is your experience with the film camera? } How many successful film cassettes do you average between camera jams? } Please quote for each TEM model (100CX, 2000FX, 3100, etc). } } Thanks, } Roseann } } Roseann Csencsits, PhD. } Donner TEM Facility Manager } Lawrence Berkeley Lab } 510-486-4548
==============================Original Headers============================== 6, 20 -- From raristau-at-ims.uconn.edu Thu Oct 12 08:21:16 2006 6, 20 -- Received: from mail2.uits.uconn.edu (mail2.uits.uconn.edu [137.99.25.204]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CDLGk3032294 6, 20 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 08:21:16 -0500 6, 20 -- Received: from [137.99.20.118] (d20h118.public.uconn.edu [137.99.20.118]) 6, 20 -- by mail2.uits.uconn.edu (8.13.6/8.11.6) with ESMTP id k9CDL9PQ019634 6, 20 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 09:21:09 -0400 6, 20 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 6, 20 -- Date: Thu, 12 Oct 2006 09:20:03 -0400 6, 20 -- Subject: Re: [Microscopy] JEOL camera question 6, 20 -- From: Roger Ristau {raristau-at-ims.uconn.edu} 6, 20 -- To: {microscopy-at-microscopy.com} 6, 20 -- Message-ID: {C153B943.17B7%raristau-at-ims.uconn.edu} 6, 20 -- In-Reply-To: {200610112114.k9BLEOoS004677-at-ns.microscopy.com} 6, 20 -- Mime-version: 1.0 6, 20 -- Content-type: text/plain; charset="US-ASCII" 6, 20 -- Content-transfer-encoding: 7bit 6, 20 -- X-UConn-MailScanner-Information: Contact UConn Help Desk 860-486-4357 for more information. 6, 20 -- X-UConn-MailScanner: Found to be clean 6, 20 -- X-UConn-MailScanner-SpamCheck: ==============================End of - Headers==============================
Roseann I have a JEOL 100 CX II, that was installed in 1982. I've been its guardian since 1988. The only time I experience camera jams is when one of my users decides to be "helpful" and reload the cassettes for me. Inevitably, one plate in the pile will have a loose corner that catches in the mechanism. As a rule, I just don't let anyone other than me and my technician load the film, and we are fine. A little care in the darkroom goes a long way. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 1, 23 -- From lcgould-at-med.cornell.edu Thu Oct 12 08:23:12 2006 1, 23 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CDNBwr003031 1, 23 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 08:23:11 -0500 1, 23 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 23 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9CDN0gA013861 1, 23 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 09:23:10 -0400 (EDT) 1, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 23 -- by mpx1.med.cornell.edu 1, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 23 -- with ESMTPA id {0J7000HS4YI1G070-at-mpx1.med.cornell.edu} for 1, 23 -- microscopy-at-microscopy.com; Thu, 12 Oct 2006 09:23:00 -0400 (EDT) 1, 23 -- Date: Thu, 12 Oct 2006 09:22:47 -0400 1, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 23 -- Subject: Re: [Microscopy] JEOL camera question 1, 23 -- In-reply-to: {200610112107.k9BL7bSs029708-at-ns.microscopy.com} 1, 23 -- Sender: lcgould-at-med.cornell.edu 1, 23 -- To: rcsencsits-at-lbl.gov, Microscopy Listserver {microscopy-at-microscopy.com} 1, 23 -- Message-id: {p06230902c153f112eaef-at-[140.251.48.23]} 1, 23 -- MIME-version: 1.0 1, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 23 -- References: {200610112107.k9BL7bSs029708-at-ns.microscopy.com} 1, 23 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.10.12.54942 ==============================End of - Headers==============================
Hi Roseann, I also have an ancient 120CX (installed 1979) as well as a newer 2011. I did have problems with the camera on the old machine occasionally (perhaps once every 10,000 plates or so), and the problem was invariably caused by my putting the film carriers in the wrong way round or not making sure the film was in the carrier properly. I find concentration does tend to drift after the first few thousand... We ran two film boxes with 50 plates, but just filling the empty one with the plates we had just taken, typically between 10 and 20 at a time. So I guess we had a problem maybe once every 500 cassette swaps. However getting the thing unstuck did also cause damage to one of the 'nails' once, and I took the camera unit out of the microscope to fix it. (Also, I have to admit, I was curious as to how it worked!) If you are having a lot of problems it may be worth it for you - it is not difficult and can be done with the machine running. It has been a while but I can give you some instructions if you want. When you have the unit out of the microscope you can feed through all of your carriers one at a time, if you like, and see if any particular ones are causing a problem. I have no problems with the 2011 camera, but I have taken less than 20 negatives in 3 years (all digital)!
Cheers
Richard
} } Dear JEOL film camera users: } } What is your experience with the film camera? } How many successful film cassettes do you average between camera jams? } Please quote for each TEM model (100CX, 2000FX, 3100, etc). } } Thanks, } Roseann } } Roseann Csencsits, PhD. } Donner TEM Facility Manager } Lawrence Berkeley Lab } 510-486-4548
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==============================Original Headers============================== 8, 31 -- From richard.beanland-at-bookham.com Thu Oct 12 08:52:47 2006 8, 31 -- Received: from mail115.messagelabs.com (mail115.messagelabs.com [195.245.231.179]) 8, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9CDqjFa021518 8, 31 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 08:52:46 -0500 8, 31 -- X-VirusChecked: Checked 8, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 8, 31 -- X-Msg-Ref: server-5.tower-115.messagelabs.com!1160660918!12701429!17 8, 31 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 8, 31 -- X-Originating-IP: [213.249.209.179] 8, 31 -- Received: (qmail 29680 invoked from network); 12 Oct 2006 13:51:00 -0000 8, 31 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 8, 31 -- by server-5.tower-115.messagelabs.com with SMTP; 12 Oct 2006 13:51:00 -0000 8, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 8, 31 -- Thu, 12 Oct 2006 14:54:06 +0100 8, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 31 -- Content-class: urn:content-classes:message 8, 31 -- MIME-Version: 1.0 8, 31 -- Content-Type: text/plain; 8, 31 -- charset="us-ascii" 8, 31 -- Subject: [Microscopy] Re: JEOL camera question 8, 31 -- Date: Thu, 12 Oct 2006 14:54:09 +0100 8, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E24316A-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 8, 31 -- X-MS-Has-Attach: 8, 31 -- X-MS-TNEF-Correlator: 8, 31 -- Thread-Topic: [Microscopy] Re: JEOL camera question 8, 31 -- Thread-Index: AcbuBePVT7HyYiPWTHe1COvrq2B6iw== 8, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 8, 31 -- To: {microscopy-at-microscopy.com} 8, 31 -- X-OriginalArrivalTime: 12 Oct 2006 13:54:06.0549 (UTC) FILETIME=[E24B9850:01C6EE05] 8, 31 -- Content-Transfer-Encoding: 8bit 8, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CDqjFa021518 ==============================End of - Headers==============================
I have been trying to help a student do Cryo-SEM on cultured T-cells - he extracts human blood, collects T-cells, cultures them in a medium containing serum and then exposes them for varying lengths of time to chemokines. After culturing the cells were washed briefly in PBS (to remove serum)then fixed in 1% glutaraldehyde in PBS for 15mins then applied to a filter where as much liquid as possible is removed immediately before freezing in nitrogen slush and carrying out Cryo-SEM. We have the problem that the cells appear in the SEM round, smooth & without psuedopodia!
We have previously done TEM on the same cells (different extraction) and had good results showing cells with interesting psuedopodia. The difference with the TEM prep was that the fixative was added to the culture medium and the cells fixed for 2hrs at room temp then overnight in the fridge.
We intend to try conventional SEM on the cells next using the initial TEM prep method then Critical Point Drying after dehydration. However I am concerned that serum will be fixed to the surface of the cells if they are not washed prior to fixation.
I would appreciate any advice on the above.
Regards Ursula ------------------------
Ursula J. Potter Centre for Electron Optical Studies (CEOS) Building 3 West 2.15 The University of Bath Claverton Down Bath BA2 7AY UK Tel: 01225 385651 Email: U.J.Potter-at-bath.ac.uk
==============================Original Headers============================== 8, 23 -- From U.J.Potter-at-bath.ac.uk Thu Oct 12 09:57:06 2006 8, 23 -- Received: from kelly.bath.ac.uk (kelly.bath.ac.uk [138.38.32.20]) 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CEv5hY004739 8, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 09:57:05 -0500 8, 23 -- Received: from mary.bath.ac.uk ([138.38.32.14] ident=mmdf) 8, 23 -- by kelly.bath.ac.uk with smtp id 1GY1zc-0006UW-4i 8, 23 -- for Microscopy-at-microscopy.com 8, 23 -- (return-path {U.J.Potter-at-bath.ac.uk} ); Thu, 12 Oct 2006 15:57:04 +0100 8, 23 -- Received: from eapc-03.campus.bath.ac.uk 8, 23 -- ( eapc-03.campus.bath.ac.uk [138.38.136.63] ) by bath.ac.uk 8, 23 -- id aa17613 for {Microscopy-at-microscopy.com} ; 12 Oct 2006 15:57 +0100 8, 23 -- Date: Thu, 12 Oct 2006 15:57:02 +0100 8, 23 -- From: Ursula Potter {U.J.Potter-at-bath.ac.uk} 8, 23 -- To: Microscopy-at-microscopy.com 8, 23 -- Subject: SEM of lymphocytes 8, 23 -- Message-ID: {24575875.1160668622-at-eapc-03.campus.bath.ac.uk} 8, 23 -- Originator-Info: login-id=mssujp; server=imaphost.bath.ac.uk 8, 23 -- X-Mailer: Mulberry/3.1.0 (Win32) 8, 23 -- MIME-Version: 1.0 8, 23 -- Content-Type: text/plain; charset=us-ascii; format=flowed 8, 23 -- Content-Transfer-Encoding: 7bit 8, 23 -- Content-Disposition: inline 8, 23 -- X-Scanner: f2dba285a45b48b55d24015519db9286d4c05056 ==============================End of - Headers==============================
I have learnt a lot about buffers used in fixative and in general about buffers in sample preparation. This is very helpful discussion. We work with mollusks (Oysters) and our oysters are reared at 860 mOSM of sea water, and a pH of near about 8.2, the cells I look at in SEM/TEM have very fine pseudopodia and the preservation of this structure in my primary concern. Could any one please suggest a buffer that works best at a pH higher than 8? After making up the buffers we measure the osmolality and adjust it, but now from the discussion I feel the need to know a buffer that can work at a higher pH.
Thank you,
Neeraj V. Gohad.
Graduate Research Assistant Department of Biological Sciences, 132 Long Hall, Clemson University, Clemson, SC-29634
==============================Original Headers============================== 7, 21 -- From ngohad-at-CLEMSON.EDU Thu Oct 12 10:43:01 2006 7, 21 -- Received: from CLEMSON.EDU (mail.clemson.edu [130.127.28.87]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CFh08I017917 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Thu, 12 Oct 2006 10:43:01 -0500 7, 21 -- Received: from X ([130.127.120.198]) 7, 21 -- by CLEMSON.EDU (8.13.6/8.13.1) with ESMTP id k9CFguVl005242 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Thu, 12 Oct 2006 11:42:56 -0400 (EDT) 7, 21 -- Message-Id: {200610121542.k9CFguVl005242-at-CLEMSON.EDU} 7, 21 -- From: "Neeraj Gohad" {ngohad-at-CLEMSON.EDU} 7, 21 -- To: {Microscopy-at-Microscopy.Com} 7, 21 -- Subject: SEM/TEM-Buffer for a higher pH 7, 21 -- Date: Thu, 12 Oct 2006 11:42:53 -0400 7, 21 -- MIME-Version: 1.0 7, 21 -- Content-Type: text/plain; 7, 21 -- charset="us-ascii" 7, 21 -- Content-Transfer-Encoding: 7bit 7, 21 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 7, 21 -- Thread-Index: AcbuFRR3p35jtat0TL27mVpV2pBoxw== 7, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 7, 21 -- X-Spam-Level: 7, 21 -- X-Scanned-By: mail mimedefang 2.56 on 130.127.28.87 ==============================End of - Headers==============================
Neeraj, In the late 1970's I worked with Mitilus and Thyone sperm in the lab of Lew Tilney. At that time the primary fixative that I used was 1% glutaraldehyde, if I remember correctly, in filtered sea water. The samples were then washed in Millonig's Phosphate Buffer and post fixed in 1% OsO4 in Phosphate Buffer -at- a pH 6.2. on ice and in the dark.
My initial reaction when I received the protocol was very strong but it worked well in this system. It may work for you also.
Pat Connelly EM Core Facility NHLBI/NIH Bethesda, MD 20892 301-496-3491
-----Original Message----- X-from: ngohad-at-CLEMSON.EDU [mailto:ngohad-at-CLEMSON.EDU] Sent: Thursday, October 12, 2006 11:52 AM To: Connelly, Patricia (NIH/NHLBI) [E]
Roseann, I just had a jam on a 100CX II caused by someone loading a plate in backwards and subsequent damage to a spring. This was the first in at least a year with 40--50 film loads and a couple thousand films exposed. I would rate the system as very reliable. Larry
rcsencsits-at-lbl.gov wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear JEOL film camera users: } } What is your experience with the film camera? } How many successful film cassettes do you average between camera jams? } Please quote for each TEM model (100CX, 2000FX, 3100, etc). } } Thanks, } Roseann } } Roseann Csencsits, PhD. } Donner TEM Facility Manager } Lawrence Berkeley Lab } 510-486-4548 } } } } } ==============================Original Headers============================== } 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 } 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) } 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 } 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 } 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) } 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 } 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) } 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) } 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; } 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) } 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) } 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} } 7, 21 -- Content-Transfer-Encoding: 7bit } 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} } 7, 21 -- Subject: JEOL camera question } 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 } 7, 21 -- To: Microscopy-at-Microscopy.Com } 7, 21 -- X-Mailer: Apple Mail (2.752.3) } 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 } 7, 21 -- X-Virus-Status: Clean } ==============================End of - Headers============================== }
-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
larry.ackerman-at-ucsf.edu
==============================Original Headers============================== 5, 30 -- From Larry.Ackerman-at-ucsf.edu Thu Oct 12 12:15:28 2006 5, 30 -- Received: from emfmcb02.ucsfmedicalcenter.org (emfmcb02.ucsfmedicalcenter.org [64.54.46.98]) 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CHFQ3A008651 5, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 12:15:27 -0500 5, 30 -- Received: from 64.54.128.152 by emfmcb02.ucsfmedicalcenter.org with 5, 30 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 5, 30 -- Thu, 12 Oct 2006 10:27:03 -0700 5, 30 -- X-Server-Uuid: 44B63953-633F-4500-B2DA-A3E478718104 5, 30 -- Received: from [128.218.123.88] ([128.218.123.88]) by 5, 30 -- exvs06.net.ucsf.edu with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 Oct 5, 30 -- 2006 10:15:19 -0700 5, 30 -- Message-ID: {452E7820.6010100-at-ucsf.edu} 5, 30 -- Date: Thu, 12 Oct 2006 10:15:12 -0700 5, 30 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 5, 30 -- Reply-to: larry.ackerman-at-ucsf.edu 5, 30 -- Organization: UCSF, NeuroAnatomy 5, 30 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 5, 30 -- X-Accept-Language: en-us, en 5, 30 -- MIME-Version: 1.0 5, 30 -- To: Microscopy-at-microscopy.com 5, 30 -- Subject: Re: [Microscopy] JEOL camera question 5, 30 -- References: {200610112111.k9BLBTYB004083-at-ns.microscopy.com} 5, 30 -- In-Reply-To: {200610112111.k9BLBTYB004083-at-ns.microscopy.com} 5, 30 -- X-OriginalArrivalTime: 12 Oct 2006 17:15:19.0319 (UTC) 5, 30 -- FILETIME=[FE3A1A70:01C6EE21] 5, 30 -- X-WSS-ID: 6930A56D1IO192507-01-01 5, 30 -- Content-Type: text/plain; 5, 30 -- charset=iso-8859-1; 5, 30 -- format=flowed 5, 30 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
On Oct 12, 2006, at 8:43 AM, ngohad-at-CLEMSON.EDU wrote:
} I have learnt a lot about buffers used in fixative and in general about } buffers in sample preparation. This is very helpful discussion. We } work } with mollusks (Oysters) and our oysters are reared at 860 mOSM of sea } water, } and a pH of near about 8.2, the cells I look at in SEM/TEM have very } fine } pseudopodia and the preservation of this structure in my primary } concern. } Could any one please suggest a buffer that works best at a pH higher } than 8? } After making up the buffers we measure the osmolality and adjust it, } but now } from the discussion I feel the need to know a buffer that can work at a } higher pH. } Dear Neeraj, I don't have access to a table of the pKs of all the Good buffers, but I seem to remember that there are some with pKs in the range you want. Failing that, the Handbook of Chemistry and Physics gives the pH for a .1N solution of NaCO3 as 8.4--of course, many cations will precipitate bicarbonate--morpholine has a pK of 8.33, several dipeptides have pKs in this range with leucylglycine at 8.28, and many alkaloids have pKs near 8.2, but explaining why you want morphine (8.21), strychnine (8.26), or codeine (8.21) may not be worth it. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Thu Oct 12 12:32:28 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CHWSdd019399 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 12:32:28 -0500 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id A3DD310A160 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 10:32:27 -0700 (PDT) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id C2F60109DCC 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 10:32:25 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200610121543.k9CFhGta018085-at-ns.microscopy.com} 4, 22 -- References: {200610121543.k9CFhGta018085-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {08661d9a0eb4733694944758640e4291-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] SEM/TEM-Buffer for a higher pH 4, 22 -- Date: Thu, 12 Oct 2006 10:36:32 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
Try skipping the glut fix. You're cryofixing the cells, so a chemical fixation should not also be needed. I assume the cells are cultured on something nice and thin and heat conductive, yes? Little metal coupons, formvar-coated TEM grids, bits of coverslip sized to the SEM stage before culturing, or the like. Rinse the cells briefly with serum-free buffer, best is the same buffer they're cultured in, just without serum. Make sure the temperature of the wash buffer is the same as the temperature of the incubation medium. Remove from the buffer and immediatey plunge into the slush nitrogen. Don't blot off the excess buffer too much -- just quick touch of the edge of the grid, etc.. There won't be much, and rapid plunging into slush nitrogen freezes fast enough that a thin layer of buffer won't interfer with good freezing. The buffer water gets vacuum sublimated during cryocoating, and if you're looking at the cells uncoated, they can still be sublimated. The conventional fixation/CPD method also works well, but I strongly suggest adding 1% monomeric tannic acid to the 1% glutaraldehyde as well as to the osmium (although OsO4 is not necessarily needed for high-resolution low-voltage SEM of cells). You are correct, the serum must be washed away however you fix the cells. Otherwise, you get these wonderful, stringy strands of serum proteins obscuring everything. Especially with chemical fixation.
Phil
} Dear All, } } I have been trying to help a student do Cryo-SEM on cultured T-cells - he } extracts human blood, collects T-cells, cultures them in a medium } containing serum and then exposes them for varying lengths of time to } chemokines. After culturing the cells were washed briefly in PBS (to } remove serum)then fixed in 1% glutaraldehyde in PBS for 15mins then applied } to a filter where as much liquid as possible is removed immediately before } freezing in nitrogen slush and carrying out Cryo-SEM. We have the problem } that the cells appear in the SEM round, smooth & without psuedopodia! } } We have previously done TEM on the same cells (different extraction) and } had good results showing cells with interesting psuedopodia. The difference } with the TEM prep was that the fixative was added to the culture medium and } the cells fixed for 2hrs at room temp then overnight in the fridge. } } We intend to try conventional SEM on the cells next using the initial TEM } prep method then Critical Point Drying after dehydration. However I am } concerned that serum will be fixed to the surface of the cells if they are } not washed prior to fixation. } } I would appreciate any advice on the above. } } Regards } Ursula } ------------------------ } } } Ursula J. Potter } Centre for Electron Optical Studies (CEOS) } Building 3 West 2.15 } The University of Bath } Claverton Down } Bath BA2 7AY } UK } Tel: 01225 385651 } Email: U.J.Potter-at-bath.ac.uk -- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Thu Oct 12 12:49:15 2006 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CHnFVO030148 4, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 12:49:15 -0500 4, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k9CIKRNC014291 4, 22 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:20:35 -0400 4, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 22 -- Thu, 12 Oct 2006 13:49:09 -0400 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f0623090ac1542d4c90e1-at-[141.209.160.249]} 4, 22 -- In-Reply-To: {200610121502.k9CF2P6F012736-at-ns.microscopy.com} 4, 22 -- References: {200610121502.k9CF2P6F012736-at-ns.microscopy.com} 4, 22 -- Date: Thu, 12 Oct 2006 13:49:07 -0400 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: Re: [Microscopy] SEM of lymphocytes 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 12 Oct 2006 17:49:09.0764 (UTC) FILETIME=[B8776C40:01C6EE26] 4, 22 -- X-CanItPRO-Stream: default 4, 22 -- X-Spam-Score: -3.4 () J_CHICKENPOX_54,L_EXCH_MF 4, 22 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Dear All, I got a mimosa as a present and I am very fascinated by this plant. I would like to study the function of the movement of the leaves with SEM and TEM. Is anybody out there who has a fixation protocoll or who did work on this plant? Any images available in the net?
Terrific little article on buffers, Good (= Good's) buffers with table or working range and pKs in wikipediat at http://stanxterm.aecom.yu.edu/wiki/index.php?page=About_buffers -mike-
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************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
Dear TEM users, We are a pathology/hospital based TEM facility equipped with a Jeol 1010 electron microscope. We would like to change to a digital camera system, since we find that wet-film technology is slow and expensive. I am hoping that there are users out there that can provide me with opinions about digital camera systems to help educate me about what they like, and what they think is important in a TEM digital camera and its associated computer [hardware/software] system.
I have never used a TEM digital camera, so I have no experience as such with any digital camera system, other than consumer digital cameras, so it's hard for me to judge the merits of a digital camera without "getting my feet wet" so to speak, and trying it out.
I'm open to suggestions, thoughts, and opinions of experienced users or any comments on this topic.
Garry Burgess
Charge Technologist Electron Microscopy Department of Pathology Winnipeg, Canada
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==============================Original Headers============================== 10, 29 -- From GBurgess-at-exchange.hsc.mb.ca Thu Oct 12 14:41:35 2006 10, 29 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJfNDI031507 10, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:41:29 -0500 10, 29 -- Received: from hscxmsmx0012.ad.wrha.mb.ca (unverified [172.16.6.178]) by 10, 29 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 10, 29 -- {B0025514934-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 10, 29 -- 12 Oct 2006 14:41:44 -0500 10, 29 -- Received: from hscxmsmx0011.ad.wrha.mb.ca ([172.16.6.192]) by 10, 29 -- hscxmsmx0012.ad.wrha.mb.ca with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 10, 29 -- Oct 2006 14:41:22 -0500 10, 29 -- Message-ID: 10, 29 -- {CEC6924DC512E24FA19F6D142677B33665B981-at-hscxmsmx0011.ad.wrha.mb.ca} 10, 29 -- Date: Thu, 12 Oct 2006 14:41:13 -0500 10, 29 -- From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} 10, 29 -- Subject: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- To: {Microscopy-at-microscopy.com} 10, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 29 -- Content-class: urn:content-classes:message 10, 29 -- X-MS-Has-Attach: 10, 29 -- X-MS-TNEF-Correlator: 10, 29 -- Thread-Topic: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- Thread-Index: AcbuNgj3H5J/RmFHRIuSUQI1vi57Cw== 10, 29 -- X-OriginalArrivalTime: 12 Oct 2006 19:41:22.0140 (UTC) 10, 29 -- FILETIME=[654679C0:01C6EE36] 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CJfNDI031507 ==============================End of - Headers==============================
We are about to scrap an Akashi 002A TEM and a 1970's vintage Balzers BAF300 freeze fracture apparatus. Before we do however, is there anyone out there who would like any specific parts or components from either of these instruments. Perhaps to keep one you have going.
You pay the cost of freight.
Regards
Allan
Allan Mitchell Otago Centre for Electron Microscopy Department of Anatomy and Structural Biology School of Medical Sciences P.O. Box 913 Dunedin New Zealand
==============================Original Headers============================== 8, 20 -- From allan.mitchell-at-stonebow.otago.ac.nz Thu Oct 12 14:54:58 2006 8, 20 -- Received: from mailhub1.otago.ac.nz (mailhub1.otago.ac.nz [139.80.64.218]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJsvuI009906 8, 20 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 14:54:58 -0500 8, 20 -- Received: from galadriel.otago.ac.nz (galadriel.otago.ac.nz [139.80.64.213]) 8, 20 -- by mailhub1.otago.ac.nz (8.13.6/8.13.6) with ESMTP id k9CJsuWS025781 8, 20 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 08:54:56 +1300 8, 20 -- Received: from allan.otago.ac.nz ([139.80.40.92]) 8, 20 -- by galadriel.otago.ac.nz with esmtp (Exim 4.50) 8, 20 -- id 1GY6ds-0007pR-Bf 8, 20 -- for microscopy-at-msa.microscopy.com; Fri, 13 Oct 2006 08:54:56 +1300 8, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 20 -- Content-Transfer-Encoding: 7bit 8, 20 -- Message-Id: {CDB10E09-AE2C-475A-8D71-A6D99D5291F6-at-stonebow.otago.ac.nz} 8, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 20 -- To: microscopy-at-msa.microscopy.com 8, 20 -- From: Allan Mitchell {allan.mitchell-at-stonebow.otago.ac.nz} 8, 20 -- Subject: Equipment being disposed of. 8, 20 -- Date: Fri, 13 Oct 2006 08:54:55 +1300 8, 20 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
Our facility is used both for research and pathology/hospital work. We have a digital camera with telescope (!) software for image collection which I find truly frustrating. You should get the highest resolution camera that you can (the pathologists here want 2K by 2K), but in my opinion you should also give the software interface high priority. I used an AMT camera at my last position and was very happy with the simplicity of use as well as the (virtually) live imaging capabilities. Most people hardly used the fluorescent screen once set up with the camera. Taking a picture was a matter of pushing an onscreen "button" and what you saw is what you got. The software was also calibrated with the microscope and didn't require manually annotating each image with the scale bar. My big complaint with that system was it was only 1K by 1K but you can easily do much better now. Kim
GBurgess-at-exchange.hsc.mb.ca wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } } Dear TEM users, } We are a pathology/hospital based TEM facility equipped with a Jeol 1010 } electron microscope. We would like to change to a digital camera system, } since we find that wet-film technology is slow and expensive. I am } hoping that there are users out there that can provide me with opinions } about digital camera systems to help educate me about what they like, } and what they think is important in a TEM digital camera and its } associated computer [hardware/software] system. } } I have never used a TEM digital camera, so I have no experience as such } with any digital camera system, other than consumer digital cameras, so } it's hard for me to judge the merits of a digital camera without } "getting my feet wet" so to speak, and trying it out. } } I'm open to suggestions, thoughts, and opinions of experienced users or } any comments on this topic. } } } Garry Burgess } } Charge Technologist } Electron Microscopy } Department of Pathology } Winnipeg, Canada } }
-- Kim Rensing Ph.D. Manager, Microscopy and Imaging Facility, University of Calgary, Health Sciences Centre B129 - 3330 Hospital Drive NW Calgary, AB, Canada T2N 4N1 O: 403-220-3488 F: 403-270-8928
==============================Original Headers============================== 4, 28 -- From krensing-at-ucalgary.ca Thu Oct 12 15:30:32 2006 4, 28 -- Received: from mr2.ucalgary.ca (mr2.ucalgary.ca [136.159.34.166]) 4, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CKUVDg021084 4, 28 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 15:30:32 -0500 4, 28 -- Received: from smtp2.ucalgary.ca (smtp2.ucalgary.ca [136.159.34.223]) 4, 28 -- by mr2.ucalgary.ca (Postfix) with ESMTP id 5A8413667D; 4, 28 -- Thu, 12 Oct 2006 14:30:31 -0600 (MDT) 4, 28 -- Received: from [192.168.0.100] ([136.159.164.171]) 4, 28 -- (authenticated (0 bits)) 4, 28 -- by smtp2.ucalgary.ca (8.11.7/8.11.6) with ESMTP id k9CKUR028214 4, 28 -- (using TLSv1/SSLv3 with cipher DHE-RSA-AES256-SHA (256 bits) verified NO); 4, 28 -- Thu, 12 Oct 2006 14:30:27 -0600 4, 28 -- Message-ID: {452EA5E0.9020107-at-ucalgary.ca} 4, 28 -- Date: Thu, 12 Oct 2006 14:30:24 -0600 4, 28 -- From: Kim Rensing {krensing-at-ucalgary.ca} 4, 28 -- Organization: Microscopy and Imaging, U of C 4, 28 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 4, 28 -- MIME-Version: 1.0 4, 28 -- To: GBurgess-at-exchange.hsc.mb.ca, microscopy-at-microscopy.com 4, 28 -- Subject: Re: [Microscopy] TEM Digital Camera - Opinions of Important Features 4, 28 -- Sought 4, 28 -- References: {200610121954.k9CJs3qs009368-at-ns.microscopy.com} 4, 28 -- In-Reply-To: {200610121954.k9CJs3qs009368-at-ns.microscopy.com} 4, 28 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 28 -- Content-Transfer-Encoding: 7bit 4, 28 -- X-UCalgary-MailScanner-Information: Please contact IT Help Desk at (403) 220-5555 for more information 4, 28 -- X-UCalgary-MailScanner: Found to be clean 4, 28 -- X-UCalgary-MailScanner-From: krensing-at-ucalgary.ca ==============================End of - Headers==============================
I'm sure you are going to get a number of answers to your question. As a manufacturer of these cameras, let me point out a few things that I think might help you. I don't want to make this into a "commercial" so I will stay away from specifics and try to provide some generic information that applies to all manufacturers. If some of this sounds too commercial, I apologize. It is not my intention.
1) TEM cameras come in two flavors: Bottom mounted and Side-mounted. The bottom-mount cameras are in general better for high-resolution imaging, while the side-mount cameras give you a better field of view. However, I would not automatically dismiss the bottom-mount cameras, as there are ways to overcome the smaller field of view (automatic image montaging), and you might find that a bottom-mount camera is better for your needs. Bottom-mount cameras are typically more expensive, so your budget will play a significant role.
2) Throughput. Most pathology departments we have talked to put a premium on efficiency so that they can work on as many cases as possible per day. If that is the case for you also, the live speed of the camera is important. It is a lot faster if you can work with the camera in a live mode, then simply push a button to snap a picture, than to work on the microscope in the traditional way (binoculars), then switch to the camera (insert in beam or lift screen), then wait for a few seconds to take a picture. If the picture didn't work out, you would have to go back to the binoculars, adjust the microscope, and start over again. A reasonable fast camera (10 frames per second) let you work on the screen computer screen directly.
3) Organization. Since digital images come cheap, users tend to take more pictures. If you have multiple users it can get quite busy on the hard disk, and you will have to implement some structure to keep users, cases, tissues, etc. apart. Here it might be advantageous to have a way of organizing your images in a database. If that is important to you, make sure that you are not missing out on that later.
4) Other instruments: Most labs have other instruments that are used for the same sample. For example, a grossing station for taking pictures of the tissue and a light microscope for color pictures. If you have that, it might be advantageous to select a system that allows information sharing and the same user interface between electron microscope and other instruments.
5) Hospital. Recently, there has been a push by pathologists to use workflow oriented software for their work, similar to what the radiologists have done for years. If this applies to you, make sure that you have an option to interface with HIS or PACS systems.
6) Reports: In most pathology labs, a technician acquires the images and hands them to the pathologist who then makes a diagnosis based on the images. This can be a critical step. Some pathologists are very particulate how the images need to look like (contrast, brightness, magnification, even paper), others work with multiple formats. Whatever is the case, you need to anticipate this. If the pathologist insists on paper prints in a certain format, the printer becomes an important part of the system. It needs to be fast, and it has to have the right tint. That's not easy to find and you need to talk to the system manufacturers for help. You might also ask the vendors about report capabilities of their software. Predefined and customized reports make it a lot easier to have the "right" format for the pathologist. A digital system, however, allows to get rid of the paper altogether. If you use a database for your images, you can probably install some software on the pathologists desktop computer, they can quickly find the images and make a diagnosis on the computer. If you think that this might be a possibility, ask the vendors about this option.
7) Support: Make sure that you can get good support for your system. Nothing is more frustrating than having to go back to film because you can't get the support you need.
8) Microscope tuning: For newer instruments, there is often an option for tuning the microscope automatically. This can help save time and make imaging easier for new staff. If that is important to you, find out if the software supports tuning the microscope.
9) Advanced features: If you require advanced features such as Tomography, make sure the system you buy supports it, or at least supports a file format that can be accepted by the third party software you use for the advanced features. In most pathology situations this is not of prime importance, though.
Ok, that's all I want to say right now. Sorry for rambling...
Michael Bode, Ph.D.
General Manager
OLYMPUS SOFT IMAGING SOLUTIONS 12596 West Bayaud Ave #300 Lakewood, CO 80228 USA Tel.: +1 (303) 234-9270 Fax.: +1 (303) 234-9271 E-mail: Mike.Bode-at-olympus-sis.com www.olympus-sis.com
-----Original Message----- X-from: GBurgess-at-exchange.hsc.mb.ca [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Thursday, October 12, 2006 13:53 To: Mike Bode
Dear TEM users, We are a pathology/hospital based TEM facility equipped with a Jeol 1010 electron microscope. We would like to change to a digital camera system, since we find that wet-film technology is slow and expensive. I am hoping that there are users out there that can provide me with opinions about digital camera systems to help educate me about what they like, and what they think is important in a TEM digital camera and its associated computer [hardware/software] system.
I have never used a TEM digital camera, so I have no experience as such with any digital camera system, other than consumer digital cameras, so it's hard for me to judge the merits of a digital camera without "getting my feet wet" so to speak, and trying it out.
I'm open to suggestions, thoughts, and opinions of experienced users or any comments on this topic.
Garry Burgess
Charge Technologist Electron Microscopy Department of Pathology Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
==============================Original Headers============================== 10, 29 -- From GBurgess-at-exchange.hsc.mb.ca Thu Oct 12 14:41:35 2006 10, 29 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJfNDI031507 10, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:41:29 -0500 10, 29 -- Received: from hscxmsmx0012.ad.wrha.mb.ca (unverified [172.16.6.178]) by 10, 29 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 10, 29 -- {B0025514934-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 10, 29 -- 12 Oct 2006 14:41:44 -0500 10, 29 -- Received: from hscxmsmx0011.ad.wrha.mb.ca ([172.16.6.192]) by 10, 29 -- hscxmsmx0012.ad.wrha.mb.ca with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 10, 29 -- Oct 2006 14:41:22 -0500 10, 29 -- Message-ID: 10, 29 -- {CEC6924DC512E24FA19F6D142677B33665B981-at-hscxmsmx0011.ad.wrha.mb.ca} 10, 29 -- Date: Thu, 12 Oct 2006 14:41:13 -0500 10, 29 -- From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} 10, 29 -- Subject: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- To: {Microscopy-at-microscopy.com} 10, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 29 -- Content-class: urn:content-classes:message 10, 29 -- X-MS-Has-Attach: 10, 29 -- X-MS-TNEF-Correlator: 10, 29 -- Thread-Topic: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- Thread-Index: AcbuNgj3H5J/RmFHRIuSUQI1vi57Cw== 10, 29 -- X-OriginalArrivalTime: 12 Oct 2006 19:41:22.0140 (UTC) 10, 29 -- FILETIME=[654679C0:01C6EE36] 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CJfNDI031507 ==============================End of - Headers==============================
==============================Original Headers============================== 32, 24 -- From Mike.Bode-at-olympus-sis.com Thu Oct 12 15:40:07 2006 32, 24 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) 32, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CKe6wK031719 32, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 15:40:06 -0500 32, 24 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 32, 24 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id k9CKl3qZ019917 32, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 22:47:06 +0200 32, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 32, 24 -- Content-class: urn:content-classes:message 32, 24 -- MIME-Version: 1.0 32, 24 -- Content-Type: text/plain; 32, 24 -- charset="us-ascii" 32, 24 -- Subject: RE: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 32, 24 -- Date: Thu, 12 Oct 2006 22:37:29 +0200 32, 24 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC9454DBA6-at-ms-s-gws.soft-imaging.net} 32, 24 -- In-Reply-To: {200610121953.k9CJrRjQ008086-at-ns.microscopy.com} 32, 24 -- X-MS-Has-Attach: 32, 24 -- X-MS-TNEF-Correlator: 32, 24 -- Thread-Topic: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 32, 24 -- Thread-Index: AcbuOBeAXmKp2X+iQ5ulvAiUn8EiPgAASjIA 32, 24 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 32, 24 -- To: {Microscopy-at-microscopy.com} 32, 24 -- Content-Transfer-Encoding: 8bit 32, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CKe6wK031719 ==============================End of - Headers==============================
Hi All, By early next year, we will have a surplus Cambridge 250 Mk3 SEM with a Link model E5431 EDS detector [all purchased in 1986] available very cheap to a good home! This SEM is still in operating condition and may be of interest to anyone who is still using an SEM of this model for spare parts, etc. Contact me if you are interested in this equipment. You would have to pay the cost of freight. Regards,
Doug Hopcroft Electron Microscope Unit, Institute of Molecular BioSciences, Massey University, Private Bag 11 222, Palmerston North, New Zealand. Phone 06 356 9099 ext.81098
==============================Original Headers============================== 4, 17 -- From d.hopcroft-at-massey.ac.nz Thu Oct 12 15:45:14 2006 4, 17 -- Received: from its-mail1.massey.ac.nz (its-mail1.massey.ac.nz [130.123.128.11]) 4, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CKjDVB006529 4, 17 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 15:45:13 -0500 4, 17 -- Received: from TUR-MM3.massey.ac.nz (tur-mm3.massey.ac.nz [130.123.128.140]) 4, 17 -- by its-mail1.massey.ac.nz (8.9.3/8.9.3) with ESMTP id JAA03815 4, 17 -- for {microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 09:45:11 +1300 (NZDT) 4, 17 -- Received: from IT041057.massey.ac.nz (Not Verified[130.123.143.35]) by TUR-MM3.massey.ac.nz with NetIQ MailMarshal 4, 17 -- id {B452ea9570001} ; Fri, 13 Oct 2006 09:45:11 +1300 4, 17 -- Message-Id: {6.2.1.2.0.20061013092853.031abb80-at-pop3.massey.ac.nz} 4, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.1.2 4, 17 -- Date: Fri, 13 Oct 2006 09:45:11 +1300 4, 17 -- To: microscopy-at-microscopy.com 4, 17 -- From: Doug Hopcroft {d.hopcroft-at-massey.ac.nz} 4, 17 -- Subject: Surplus Cambridge 250 Mk3 SEM. 4, 17 -- Mime-Version: 1.0 4, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Can someone suggest help with sections falling off of Ni grids during Immuno gold staining? Grids are clean and sections are dried overnight to 3 days prior to staining. I have not yet tried a pap pen to adhere the sections...but I'm looking at my "Coat-Quick "G" grid coating pen and beginning to ask.... Why not? Any reasons not to use it?
Thanks, Linda
Linda M. Fox Loyola University Stritch School of Medicine Core Imaging Facility 2160 S. First Ave. Maywood, Il 60153 Bld. 102 Room 0617 1-708-216-3395 lfox1-at-lumc.edu
==============================Original Headers============================== 7, 17 -- From lfox1-at-lumc.edu Thu Oct 12 16:14:08 2006 7, 17 -- Received: from GWPRIMARY.luhs.org (gw5gate.lumc.edu [147.126.200.222]) 7, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CLE8IU021017 7, 17 -- for {Microscopy-at-Microscopy.Com} ; Thu, 12 Oct 2006 16:14:08 -0500 7, 17 -- Received: from Primary-MTA by GWPRIMARY.luhs.org 7, 17 -- with Novell_GroupWise; Thu, 12 Oct 2006 16:14:14 -0500 7, 17 -- Message-Id: {s52e69d6.030-at-GWPRIMARY.luhs.org} 7, 17 -- X-Mailer: Novell GroupWise Internet Agent 6.5.4 7, 17 -- Date: Thu, 12 Oct 2006 16:13:36 -0500 7, 17 -- From: "Linda Fox" {lfox1-at-lumc.edu} 7, 17 -- To: {Microscopy-at-Microscopy.Com} 7, 17 -- Subject: ICC gold and pap pens? 7, 17 -- Mime-Version: 1.0 7, 17 -- Content-Type: text/plain; charset=US-ASCII 7, 17 -- Content-Disposition: inline 7, 17 -- Content-Transfer-Encoding: 8bit 7, 17 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CLE8IU021017 ==============================End of - Headers==============================
There's also a useful booklet from Calbiochem called (you guessed it) Buffers: A guide for the preparation and use of buffers in biological systems. I have the "new" 1995 edition, but it has all the Good buffers, recipes, comments about other issues such as cation chelation, notes on protein pI, etc. Rosemary
} From: mike.reedy-at-cellbio.duke.edu } Reply-To: mike.reedy-at-cellbio.duke.edu } Date: Thu, 12 Oct 2006 13:32:18 -0500 } To: rosemary.white-at-csiro.au } Subject: [Microscopy] Re: SEM/TEM-Buffer for a higher pH } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Terrific little article on buffers, Good (= Good's) buffers with } table or working range and pKs in wikipediat at } http://stanxterm.aecom.yu.edu/wiki/index.php?page=About_buffers } -mike- } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } } } On Oct 12, 2006, at 8:43 AM, ngohad-at-CLEMSON.EDU wrote: } } } } } I have learnt a lot about buffers used in fixative and in general about } } } buffers in sample preparation. This is very helpful discussion. We } } } work } } } with mollusks (Oysters) and our oysters are reared at 860 mOSM of sea } } } water, } } } and a pH of near about 8.2, the cells I look at in SEM/TEM have very } } } fine } } } pseudopodia and the preservation of this structure in my primary } } } concern. } } } Could any one please suggest a buffer that works best at a pH higher } } } than 8? } } } After making up the buffers we measure the osmolality and adjust it, } } } but now } } } from the discussion I feel the need to know a buffer that can work at a } } } higher pH. } } } } } Dear Neeraj, } } I don't have access to a table of the pKs of all the Good buffers, but } } I seem to remember that there are some with pKs in the range you want. } } Failing that, the Handbook of Chemistry and Physics gives the pH for a } } .1N solution of NaCO3 as 8.4--of course, many cations will precipitate } } bicarbonate--morpholine has a pK of 8.33, several dipeptides have pKs } } in this range with leucylglycine at 8.28, and many alkaloids have pKs } } near 8.2, but explaining why you want morphine (8.21), strychnine } } (8.26), or codeine (8.21) may not be worth it. } } Yours, } } Bill Tivol, PhD } } EM Scientist and Manager } } Cryo-Electron Microscopy Facility } } Broad Center, Mail Code 114-96 } } California Institute of Technology } } Pasadena CA 91125 } } (626) 395-8833 } } tivol-at-caltech.edu } } } } } } ==============================Original Headers============================== } } 4, 22 -- From tivol-at-caltech.edu Thu Oct 12 12:32:28 2006 } } 4, 22 -- Received: from outgoing-mail.its.caltech.edu } } (outgoing-mail.its.caltech.edu [131.215.239.19]) } } 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k9CHWSdd019399 } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 } } 12:32:28 -0500 } } 4, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) } } 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id A3DD310A160 } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 } } 10:32:27 -0700 (PDT) } } 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu } } [131.215.2.133]) } } 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id C2F60109DCC } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 12 Oct 2006 } } 10:32:25 -0700 (PDT) } } 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) } } 4, 22 -- In-Reply-To: {200610121543.k9CFhGta018085-at-ns.microscopy.com} } } 4, 22 -- References: {200610121543.k9CFhGta018085-at-ns.microscopy.com} } } 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } } 4, 22 -- Message-Id: {08661d9a0eb4733694944758640e4291-at-caltech.edu} } } 4, 22 -- Content-Transfer-Encoding: 7bit } } 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} } } 4, 22 -- Subject: Re: [Microscopy] SEM/TEM-Buffer for a higher pH } } 4, 22 -- Date: Thu, 12 Oct 2006 10:36:32 -0700 } } 4, 22 -- To: microscopy-at-msa.microscopy.com } } 4, 22 -- X-Mailer: Apple Mail (2.624) } } 4, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 } } ==============================End of - Headers============================== } } } -- } -mike reedy- } } ************************ } Michael K. Reedy, M.D. } Duke Univ. Med. Center } Dept. Cell Biology, Box 3011 (for U.S. Mail) } 458 Alex Sands Bldg, Research Dr (courier) } Durham, NC 27710 } } Office 919-668-2534 } Lab 919-684-5674 } Fax 919-681-9929 } mike.reedy-at-cellbio.duke.edu } http://note.cellbio.duke.edu/Faculty/Research/Reedy.html } } ==============================Original Headers============================== } 6, 22 -- From mike.reedy-at-cellbio.duke.edu Thu Oct 12 13:29:29 2006 } 6, 22 -- Received: from porthos.duhs.duke.edu (porthos.duhs.duke.edu } [152.16.199.201]) } 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k9CITTBg019570 } 6, 22 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 13:29:29 -0500 } 6, 22 -- Received: from cellbio.duke.edu (voice.cellbio.duke.edu } [152.16.14.127]) } 6, 22 -- by porthos.duhs.duke.edu (8.13.4/8.13.4) with ESMTP id } k9CIT6E91249430 } 6, 22 -- (version=TLSv1/SSLv3 cipher=DES-CBC3-SHA bits=168 verify=NO); } 6, 22 -- Thu, 12 Oct 2006 14:29:06 -0400 } 6, 22 -- Received: from [152.16.192.164] (HELO [152.16.6.135]) } 6, 22 -- by cellbio.duke.edu (CommuniGate Pro SMTP 4.1.8) } 6, 22 -- with ESMTP id 4263121; Thu, 12 Oct 2006 14:29:05 -0400 } 6, 22 -- Mime-Version: 1.0 } 6, 22 -- Message-Id: {p06210218c15439705d76-at-[152.16.6.135]} } 6, 22 -- In-Reply-To: {200610121736.k9CHarYO025302-at-ns.microscopy.com} } 6, 22 -- References: {200610121736.k9CHarYO025302-at-ns.microscopy.com} } 6, 22 -- Date: Thu, 12 Oct 2006 14:29:00 -0400 } 6, 22 -- To: tivol-at-caltech.edu } 6, 22 -- From: Mike Reedy {mike.reedy-at-cellbio.duke.edu} } 6, 22 -- Subject: [Microscopy] Re: SEM/TEM-Buffer for a higher pH } 6, 22 -- Cc: "Microscopy Listserver" {microscopy-at-microscopy.com} } 6, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 6, 22 -- X-Scanned-By: MIMEDefang 2.51 on 152.16.199.201 } ==============================End of - Headers============================== }
==============================Original Headers============================== 5, 22 -- From Rosemary.White-at-csiro.au Thu Oct 12 16:45:35 2006 5, 22 -- Received: from act-MTAout2.csiro.au (act-MTAout2.csiro.au [150.229.7.38]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CLjX99032008 5, 22 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 16:45:34 -0500 5, 22 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=TNdkGIbxJWsUaiiJzdToDEbWNEpXyxE/knlrwvDnwxsnjF9Rrvx0u+LQVGUXNx7VCnlObtIeSpSsvtb/YeLNPmi5d0Su+/3VudyufDe/ffRD6axuQ93xMWNr/0et2Aia; 5, 22 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 5, 22 -- by act-MTAout2.csiro.au with ESMTP; 13 Oct 2006 07:44:40 +1000 5, 22 -- X-IronPort-AV: i="4.09,301,1157292000"; 5, 22 -- d="scan'208"; a="115407943:sNHT29599632" 5, 22 -- Received: from [152.83.167.45] ([152.83.167.45]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 5, 22 -- Fri, 13 Oct 2006 07:45:22 +1000 5, 22 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 5, 22 -- Date: Fri, 13 Oct 2006 07:47:21 +1000 5, 22 -- Subject: Re: [Microscopy] Re: SEM/TEM-Buffer for a higher pH 5, 22 -- From: Rosemary White {Rosemary.White-at-csiro.au} 5, 22 -- To: {microscopy-at-microscopy.com} 5, 22 -- Message-ID: {C154F509.197E7%Rosemary.White-at-csiro.au} 5, 22 -- In-Reply-To: {200610121832.k9CIWIgZ024019-at-ns.microscopy.com} 5, 22 -- Mime-version: 1.0 5, 22 -- Content-type: text/plain; charset="US-ASCII" 5, 22 -- Content-transfer-encoding: 7bit 5, 22 -- X-OriginalArrivalTime: 12 Oct 2006 21:45:22.0676 (UTC) FILETIME=[B82E2B40:01C6EE47] ==============================End of - Headers==============================
Considering that plants have 'rigid' cell walls, its amazing how they achieve sometimes very rapid movement!
I came across a paper on Mimosa pudica a couple of years ago, in relation to the anatomy of the motor organs at the bases of the prtiole, rachis and pinnule. The paper focuses on the vascular tissue, but its a start:
Fleurat-Lessard and Bonnemain (1978) Structural and ultrastructural charactersitics of the vascular apparatus of the sensitive plant (Mimosa pudica L.) Protoplasma 94: 127-143
Hope this helps,
Mark
Mark Talbot CSIRO Plant Industry GPO Box 1600 Canberra, ACT 2601 mark.talbot-at-csiro.au ph: 02-6246 5256 fax: 02-6246 5334
==============================Original Headers============================== 8, 30 -- From mark.talbot-at-csiro.au Thu Oct 12 17:48:11 2006 8, 30 -- Received: from act-MTAout6.csiro.au (act-MTAout6.csiro.au [150.229.7.43]) 8, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CMm9Rq020090 8, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 17:48:10 -0500 8, 30 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=n2eEEmLInUTTCKsjzABbDUEd2aEiZMnTnOoMssiF2gFPo2LzK8F7safJNb+TdJVHWVhCM3Rcv8KCrohyKgsb0JX2YbyOoKSwWklBFmQKIqz+9NGwULOdxPlCWR9DGI1X; 8, 30 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 8, 30 -- by act-MTAout6.csiro.au with ESMTP; 13 Oct 2006 08:47:16 +1000 8, 30 -- X-IronPort-AV: i="4.09,301,1157292000"; 8, 30 -- d="scan'208"; a="115417095:sNHT24606672" 8, 30 -- Received: from exvicn1-mel.nexus.csiro.au ([138.194.3.60]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 8, 30 -- Fri, 13 Oct 2006 08:47:59 +1000 8, 30 -- Received: from exnswn1-syd.nexus.csiro.au ([130.155.3.31]) by exvicn1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 8, 30 -- Fri, 13 Oct 2006 08:47:59 +1000 8, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 30 -- content-class: urn:content-classes:message 8, 30 -- MIME-Version: 1.0 8, 30 -- Content-Type: text/plain; 8, 30 -- charset="Windows-1252" 8, 30 -- Subject: Re: Mimosa questions... 8, 30 -- Date: Fri, 13 Oct 2006 08:47:58 +1000 8, 30 -- Message-ID: {BF94750921492E4AA5B825026FCE6A2401489FCE-at-exnswn1-syd.nexus.csiro.au} 8, 30 -- X-MS-Has-Attach: 8, 30 -- X-MS-TNEF-Correlator: 8, 30 -- Thread-Topic: Re: Mimosa questions... 8, 30 -- Thread-Index: AcbuTzeBN4bi+CD3Q9SYs6jcdEe4Tw== 8, 30 -- From: {mark.talbot-at-csiro.au} 8, 30 -- To: {Microscopy-at-microscopy.com} 8, 30 -- X-OriginalArrivalTime: 12 Oct 2006 22:47:59.0394 (UTC) FILETIME=[775BD820:01C6EE50] 8, 30 -- Content-Transfer-Encoding: 8bit 8, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CMm9Rq020090 ==============================End of - Headers==============================
Michael, I googled neslab and got www.neslab.com . They're apparently part of the Thermo family now (as is nearly everyone in the EM accessories industry). There were also a huge number of used Neslab chillers listed. My customers have generally had good luck with their products and I've found their service to be quite responsive and helpful (along with Haskris).
No stake, just like their stuff.
Ken Converse owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: mdelann1-at-jhmi.edu [mailto:mdelann1-at-jhmi.edu] Sent: Wednesday, October 11, 2006 9:39 PM To: kenconverse-at-qualityimages.biz
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Email: mdelann1-at-jhmi.edu Name: Michael Delannoy
Organization: Assistant Director Imaging Facility JHSM
Title-Subject: [Filtered] chillers
Question: Hello I need a few companies who would sell used/ refurbished water chillers/recirculators. I need a supply for my LEO 1530 FESEM Does anyone know the heat capaciy required? Does anyone know the Neslabs website or contact info? Thank You M Delannoy 410 955-1365
==============================Original Headers============================== 6, 12 -- From zaluzec-at-microscopy.com Wed Oct 11 20:35:33 2006 6, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 6, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9C1ZVoF003898 6, 12 -- for {microscopy-at-microscopy.com} ; Wed, 11 Oct 2006 20:35:33 -0500 6, 12 -- Mime-Version: 1.0 6, 12 -- X-Sender: (Unverified) 6, 12 -- Message-Id: {p06110405c1534c51302b-at-[206.69.208.22]} 6, 12 -- Date: Wed, 11 Oct 2006 20:35:30 -0500 6, 12 -- To: microscopy-at-microscopy.com 6, 12 -- From: mdelann1-at-jhmi.edu (by way of MicroscopyListserver) 6, 12 --
Chillers....a really interesting area and quite surprising.
The Thermo units I think across the board have 0.5 gallon reservoirs. If your flow rate is really low, this should not be an issue. I think like .5GPH or so which is what the FEI systems use. They can do this because their systems dump heat into the area where the SEM and expansion racks are located. Zeiss takes a different approach to cooling. They use the Haskris chillers with 5 gallon reservoirs and add some additional interesting options. A key one is the hot gas bypass.
Myself and others have second-guessed the Zeiss folks and come out with the short end of the stick. The hot gas bypass is a feature that dumps hot refrigerant back into the reservoir thus causing the compressor to not shut off. The net results of this are that the compressor rarely shuts off and water temperature at high (30GPM or H, not sure) flow is constant. Also, the chiller takes most all of the SEM plinth heat out to where the chiller is. So, if located in different rooms, the SEM room is easy to cool.
Another factor is that the compressor does not short cycle. This is fatal to the compressor if it does short cycle--on and off in a short period of time. The start relay will eventually burn out and require replacement of the whole compressor unit if this happens. Short cycling happens when the temperature setting has hysterisis. E. g., 65F - 67F. When the water reaches 67F, the compressor starts and cools the water to 65F then shuts off. Depending on the heat load, this can happen very frequently. With low demand systems like FEI, the compressors see the same thing but seem to take it in stride. The Tecumseh compressors in the Haskris chillers apparently do not like this.
Anyway, Haskris has excellent customer service from my experience and the Thermo units are also good for their applications. So, I guess that it just "depends."
Disclaimer: blah, blah.
gary g.
At 04:32 PM 10/12/2006, you wrote:
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==============================Original Headers============================== 14, 20 -- From gary-at-gaugler.com Thu Oct 12 19:16:37 2006 14, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9D0GbVl026305 14, 20 -- for {microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 19:16:37 -0500 14, 20 -- Received: (qmail 14988 invoked from network); 12 Oct 2006 17:16:32 -0700 14, 20 -- Received: by simscan 1.1.0 ppid: 14976, pid: 14986, t: 0.2404s 14, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 14, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 14, 20 -- by qsmtp1 with SMTP; 12 Oct 2006 17:16:32 -0700 14, 20 -- Message-Id: {7.0.1.0.2.20061012165435.0259ae00-at-gaugler.com} 14, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 20 -- Date: Thu, 12 Oct 2006 17:16:39 -0700 14, 20 -- To: kenconverse-at-qualityimages.biz 14, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 14, 20 -- Subject: Re: [Microscopy] RE: viaWWW:water chillers 14, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 14, 20 -- In-Reply-To: {200610122332.k9CNWUnj010065-at-ns.microscopy.com} 14, 20 -- References: {200610122332.k9CNWUnj010065-at-ns.microscopy.com} 14, 20 -- Mime-Version: 1.0 14, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Our newer EFTEM has been dead for a few weeks, so I got our venerable old Zeiss 10/A going. This meant loading in old film from 2001, cleaning out the darkroom, making up old (and black) D-19, and generally stepping back into the past. Amazingly, the images are great! We've been very happy with them; giddy, even! However, our stash of old film is nearly gone, and I remember the rant on this list a couple of years ago about the new, "improved" Kodak 4489. Indeed, one user bought new film (no instruction sheet included!) and new D-19 (new plastic packaging), and we are terribly unhappy with the results. I went back through the List's archives, and we tried all the tricks and patiently tried all kinds of dilutions of developer, development times, exposures, etc., all with poor results. We're not getting uneven development; our agitation seems to be fine. It seems that the response curve of the new film is more linear instead of the traditional S-shape, and we're not getting good detail in the darker blacks and whiter whites. And the film seems to have a brown cast!
I would be interested in hearing how any of you have been dealing with this, especially within the past year.
Mahalo, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
==============================Original Headers============================== 7, 19 -- From tina-at-pbrc.hawaii.edu Thu Oct 12 19:30:41 2006 7, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9D0UfVV007093 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 19:30:41 -0500 7, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 7, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k9D0UbAk013635 7, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 14:30:38 -1000 (HST) 7, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k9D0Ua2g013632 7, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 12 Oct 2006 14:30:37 -1000 (HST) 7, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 7, 19 -- Date: Thu, 12 Oct 2006 14:30:36 -1000 (HST) 7, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 7, 19 -- X-Sender: tina-at-halia 7, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 7, 19 -- Subject: Kodak 4489 film 7, 19 -- Message-ID: {Pine.GSO.4.21.0610121418500.13499-100000-at-halia} 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
I am doing fundamental research with a Megaview III digital camera from Olympus-SIS imaging (side-mounted camera). I find this material just fantastic. No need to observe on the fluorescent screen, with several advantages: - You can take a picture immediately: find a field, take a picture. 1 second later you have your results and can adjust if needed. - the camera being much more sensitive that your eyes on the fluorescent screen, you can reduce the dose of the beam, meaning less damage to your material and better contrast. - Your can organize your results immediately: no need to take notes, numerate your negative, calculate the magnification...
In addition to the camera comes a great software which allows you to organize very efficiently your pictures in databases and reports. The software also allows all sorts of analysis and calculations from your pictures. You want more? Well actually you get more, since the technical assistance is really efficient too!
Important things to consider are the resolution of the camera (although I wonder why a pathologist would need the highest resolution possible - it is an open question, i am not pathologist) and the horizontal transfer of the informations between the camera software and other software used in your hospital
Regards,
Stephane
----- Original Message ---- X-from: "GBurgess-at-exchange.hsc.mb.ca" {GBurgess-at-exchange.hsc.mb.ca} To: nizets2-at-yahoo.com Sent: Thursday, October 12, 2006 9:53:09 PM
Dear TEM users, We are a pathology/hospital based TEM facility equipped with a Jeol 1010 electron microscope. We would like to change to a digital camera system, since we find that wet-film technology is slow and expensive. I am hoping that there are users out there that can provide me with opinions about digital camera systems to help educate me about what they like, and what they think is important in a TEM digital camera and its associated computer [hardware/software] system.
I have never used a TEM digital camera, so I have no experience as such with any digital camera system, other than consumer digital cameras, so it's hard for me to judge the merits of a digital camera without "getting my feet wet" so to speak, and trying it out.
I'm open to suggestions, thoughts, and opinions of experienced users or any comments on this topic.
Garry Burgess
Charge Technologist Electron Microscopy Department of Pathology Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
==============================Original Headers============================== 10, 29 -- From GBurgess-at-exchange.hsc.mb.ca Thu Oct 12 14:41:35 2006 10, 29 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJfNDI031507 10, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:41:29 -0500 10, 29 -- Received: from hscxmsmx0012.ad.wrha.mb.ca (unverified [172.16.6.178]) by 10, 29 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 10, 29 -- {B0025514934-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 10, 29 -- 12 Oct 2006 14:41:44 -0500 10, 29 -- Received: from hscxmsmx0011.ad.wrha.mb.ca ([172.16.6.192]) by 10, 29 -- hscxmsmx0012.ad.wrha.mb.ca with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 10, 29 -- Oct 2006 14:41:22 -0500 10, 29 -- Message-ID: 10, 29 -- {CEC6924DC512E24FA19F6D142677B33665B981-at-hscxmsmx0011.ad.wrha.mb.ca} 10, 29 -- Date: Thu, 12 Oct 2006 14:41:13 -0500 10, 29 -- From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} 10, 29 -- Subject: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- To: {Microscopy-at-microscopy.com} 10, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 29 -- Content-class: urn:content-classes:message 10, 29 -- X-MS-Has-Attach: 10, 29 -- X-MS-TNEF-Correlator: 10, 29 -- Thread-Topic: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- Thread-Index: AcbuNgj3H5J/RmFHRIuSUQI1vi57Cw== 10, 29 -- X-OriginalArrivalTime: 12 Oct 2006 19:41:22.0140 (UTC) 10, 29 -- FILETIME=[654679C0:01C6EE36] 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CJfNDI031507 ==============================End of - Headers==============================
==============================Original Headers============================== 24, 20 -- From nizets2-at-yahoo.com Fri Oct 13 03:17:53 2006 24, 20 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.91.145]) 24, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9D8Hrxn015799 24, 20 -- for {microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 03:17:53 -0500 24, 20 -- Received: (qmail 80657 invoked by uid 60001); 13 Oct 2006 08:17:53 -0000 24, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 24, 20 -- s=s1024; d=yahoo.com; 24, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:MIME-Version:Content-Type:Content-Transfer-Encoding; 24, 20 -- b=JchCkauFp84T1QQQjnrkI0BbEttrgkjOKqR/kR1Xjd72lz31qZD0EBCzIXiHEs7nqHw7WPipEGRllnCs6F0Fbii2a6BEl8xrQVxr7+hwlluXAcUPtutOzEGcoqtrJgC77da90Z+PdMfwkNuyNkKc77q0aWN4ueZLQOSzbXUKNuo= ; 24, 20 -- Message-ID: {20061013081753.80655.qmail-at-web37413.mail.mud.yahoo.com} 24, 20 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Fri, 13 Oct 2006 01:17:53 PDT 24, 20 -- Date: Fri, 13 Oct 2006 01:17:53 -0700 (PDT) 24, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 24, 20 -- Subject: Re: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 24, 20 -- To: GBurgess-at-exchange.hsc.mb.ca 24, 20 -- Cc: microscopy-at-microscopy.com 24, 20 -- MIME-Version: 1.0 24, 20 -- Content-Type: text/plain; charset=ascii 24, 20 -- Content-Transfer-Encoding: 8bit 24, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9D8Hrxn015799 ==============================End of - Headers==============================
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Email: mark.grimson-at-ttu.edu Name: Mark Grimson
Organization: Texas Tech University
Title-Subject: [Filtered] Horizontal lines on a Gatan slow-scan TEM camera
Question: Hello. We recently recieved an H-8100 with a bottom mount Gatan slow-scan CCD cooled camera Model 794 multi-scan camera). Before it left its old location the camera was functioning normally, but when it arrived here, it developed some problems, in particular, horizontal lines running across the acquisition screen in all 3 modes. The image that is projected on the camera can be observed between the lines. Has anyone had experience with this problem before. I would appreciate any suggestions before sending the camera back to Gatan for repairs. Thanks for any advice or comments. Mark
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Email: jvtaylo-at-emory.edu Name: Jeannette Taylor
Organization: IM&MF, Emory University
Title-Subject: [Filtered] Surplus DS130 LaB6 SEM
Question: Dear All, we have a Topcon DS130 SEM, with LaB6 filament, in operating condition, available very cheap to interested parties. This instrument has in-lens imaging as well as conventional below-the-lens imaging. Other features as well. Please inquire.
Thanks, Jeannette Taylor
Jeannette Taylor, Technologist II IM&MF, Emory University (404) 712-8674 jvtaylo-at-emory.edu
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Title-Subject: [Filtered] Electron Microscopist Salary
Question: I am trying to hire an electron microscopist who has 3-5 years of research and technical experience using TEM. I am having difficulties getting past HR who want to pay this techmician far less than I think reasonable for the position. Can anyone provide me with salary ranges suggested for RA-I type positions, in a university setting, for electron microspists.
I never had any problems with the new 4489 film. I developed it in D-19, diluted 1:2, for 4 min 15 seconds at 70 F. Gentle agitation for the first 30 seconds, then for 5 seconds every 30 seconds. Rinse, fix and wash as usual. They only difference is that I make my D-19 (and all of my other B&W developers) from scratch. Not from the Kodak package, I have the formula, each of the raw ingredients and I weigh them out and dissolve them in the proper order in distilled water. I am surprised you got good results from old, black D-19!
Geoff
tina-at-pbrc.hawaii.edu wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 9, 34 -- From mcauliff-at-umdnj.edu Fri Oct 13 09:03:10 2006 9, 34 -- Received: from zix01.umdnj.edu (zix01.UMDNJ.EDU [130.219.34.124]) 9, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DE39Ls020998 9, 34 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 09:03:09 -0500 9, 34 -- Received: from zix01.umdnj.edu (ZixVPM [127.0.0.1]) 9, 34 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id AF22A1C328C 9, 34 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 10:03:08 -0400 (EDT) 9, 34 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 9, 34 -- by zix01.umdnj.edu (Proprietary) with ESMTP id 29B05A7B4A 9, 34 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 10:03:07 -0400 (EDT) 9, 34 -- Received: from ([130.219.34.131]) 9, 34 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.38457316; 9, 34 -- Fri, 13 Oct 2006 09:53:54 -0400 9, 34 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 9, 34 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 9, 34 -- id {0J7200B01U9ESQ-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 9, 34 -- for microscopy-at-msa.microscopy.com; Fri, 13 Oct 2006 09:53:54 -0400 (EDT) 9, 34 -- Received: from [127.0.0.1] ([10.138.2.123]) 9, 34 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 9, 34 -- 2004)) with ESMTP id {0J72008BDUKN2C-at-Polaris.umdnj.edu} ; Fri, 9, 34 -- 13 Oct 2006 09:53:12 -0400 (EDT) 9, 34 -- Date: Fri, 13 Oct 2006 09:54:22 -0400 9, 34 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 9, 34 -- Subject: Re: [Microscopy] Kodak 4489 film 9, 34 -- In-reply-to: {200610130032.k9D0WRhh009323-at-ns.microscopy.com} 9, 34 -- To: tina-at-pbrc.hawaii.edu, MicroscopyListserver {microscopy-at-msa.microscopy.com} 9, 34 -- Message-id: {452F9A8E.7050603-at-umdnj.edu} 9, 34 -- MIME-version: 1.0 9, 34 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 9, 34 -- Content-transfer-encoding: 7BIT 9, 34 -- X-Accept-Language: en-us, en 9, 34 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 9, 34 -- Gecko/20040804 Netscape/7.2 (ax) 9, 34 -- References: {200610130032.k9D0WRhh009323-at-ns.microscopy.com} ==============================End of - Headers==============================
Hi Tina, I use new improved Kodak Estar 4489. I had to change safety light in my dark room from sodium light to a red filter because this film is apparently sensitive to sodium light (according to the manufacturer insert). Dorota
==============================Original Headers============================== 1, 22 -- From wadowska-at-avcn1.novell.upei.ca Fri Oct 13 09:43:18 2006 1, 22 -- Received: from mx2.upei.ca (magellanic.cs.upei.ca [137.149.3.22]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DEhI1t010869 1, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 09:43:18 -0500 1, 22 -- Received: from [137.149.64.250] (helo=acad1.cs.upei.ca) 1, 22 -- by mx2.upei.ca with esmtp (Exim 4.50 #1 (Debian)) 1, 22 -- id 1GYOFq-0005EI-Se 1, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 13 Oct 2006 11:43:19 -0300 1, 22 -- Received: from AVCN1/SpoolDir by acad1.cs.upei.ca (Mercury 1.48); 1, 22 -- 13 Oct 06 11:43:18 -0300 1, 22 -- Received: from SpoolDir by AVCN1 (Mercury 1.48); 13 Oct 06 11:42:45 -0300 1, 22 -- From: "Dorota Wadowska" {wadowska-at-upei.ca} 1, 22 -- Organization: University of P.E.I. 1, 22 -- To: microscopy-at-msa.microscopy.com 1, 22 -- Date: Fri, 13 Oct 2006 11:36:08 -0400 1, 22 -- MIME-Version: 1.0 1, 22 -- Content-type: text/plain; charset=US-ASCII 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- Subject: Re: Kodak 4489 film 1, 22 -- Message-ID: {452F7A27.15586.6A703A-at-localhost} 1, 22 -- Priority: normal 1, 22 -- X-mailer: Pegasus Mail for Win32 (v3.12c) ==============================End of - Headers==============================
Microscopy Today magazine published a salary survey in January 2005, attached.* I would add a couple of percent to reflect salary increases between late 2004, when this data was submitted, and now. We'll be doing another survey in about six months.
Ron Anderson, Editor Microscopy Today
Subscribe at http://www.microscopy-today.com, free in North America. *Attached in response to Karen
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==============================Original Headers============================== 5, 19 -- From microscopytoday-at-tampabay.rr.com Fri Oct 13 10:00:30 2006 5, 19 -- Received: from ms-smtp-06.tampabay.rr.com (ms-smtp-06.tampabay.rr.com [65.32.5.136]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DF0Ubw021753 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Fri, 13 Oct 2006 10:00:30 -0500 5, 19 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 5, 19 -- by ms-smtp-06.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k9DF0RBf011246 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Fri, 13 Oct 2006 11:00:29 -0400 (EDT) 5, 19 -- Message-ID: {452FAA09.5050305-at-tampabay.rr.com} 5, 19 -- Date: Fri, 13 Oct 2006 11:00:25 -0400 5, 19 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 5, 19 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: Listserver {Microscopy-at-Microscopy.Com} 5, 19 -- Subject: Re: [Microscopy] viaWWW: Electron Microscopist Salary 5, 19 -- References: {200610131354.k9DDslTx002930-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200610131354.k9DDslTx002930-at-ns.microscopy.com} 5, 19 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 19 -- Content-Transfer-Encoding: 7bit 5, 19 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
You may use any composite video monitor (9" B&W security monitor), given that H-600 takes composite video- I am not 100% certain about H-600, but most data monitors like that use standard composite video 1V p-p; 75 Ohm. Worth checking out IMHO.
Vitaly Feingold SIA 2773 Heath Line Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com ----- Original Message ----- X-from: {john.brealey-at-imvs.sa.gov.au} To: {vitalylazar-at-att.net} Sent: Wednesday, October 11, 2006 10:03 PM
Dear Colleagues...
We're trying to process skin tissue samples for TEM. Despite several attempts with different protocols, we couldn't achieve fully satisfactory results.Has anybody experienced skin tissue processing? Are there "tips and tricks" for processing skin tissue? Thank you in advance...
Dr. Nejat Yilmaz
==============================Original Headers============================== 8, 28 -- From nyilmaz-at-mersin.edu.tr Fri Oct 13 14:17:22 2006 8, 28 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DJHLFG015879 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 14:17:22 -0500 8, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 8, 28 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 25FF94508A 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 22:32:59 +0300 (EEST) 8, 28 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr 8, 28 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 8, 28 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 8, 28 -- with ESMTP id K66xSD0aheDH for {Microscopy-at-microscopy.com} ; 8, 28 -- Fri, 13 Oct 2006 22:32:56 +0300 (EEST) 8, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 8, 28 -- by mail.mersin.edu.tr (Postfix) with SMTP id 21AA6450AE 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 22:32:56 +0300 (EEST) 8, 28 -- To: {Microscopy-at-microscopy.com} 8, 28 -- Subject: Skin processing 8, 28 -- From: {nyilmaz-at-mersin.edu.tr} 8, 28 -- Date: Fri, 13 Oct 2006 22:32:56 EEST 8, 28 -- Reply-To: {nyilmaz-at-mersin.edu.tr} 8, 28 -- Errors-To: {nyilmaz-at-mersin.edu.tr} 8, 28 -- X-Priority: 3 (Normal) 8, 28 -- X-Originating-Ip: [85.104.168.63] 8, 28 -- X-Mailer: NOCC v0.9.5 8, 28 -- Content-Type: text/plain; 8, 28 -- charset="ISO-8859-9" 8, 28 -- Content-Transfer-Encoding: 8bit 8, 28 -- Message-Id: {20061013193256.21AA6450AE-at-mail.mersin.edu.tr} ==============================End of - Headers==============================
What kind of skin are you processing? We do mouse skin routinely and successfully for TEM. I usually embed in Spurrs resin and do long infiltration steps.
Lesley
Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322 -----Original Message----- X-from: nyilmaz-at-mersin.edu.tr [mailto:nyilmaz-at-mersin.edu.tr] Sent: Friday, October 13, 2006 3:25 PM To: lesley.bechtold-at-jax.org
Dear Colleagues...
We're trying to process skin tissue samples for TEM. Despite several attempts with different protocols, we couldn't achieve fully satisfactory results.Has anybody experienced skin tissue processing? Are there "tips and tricks" for processing skin tissue? Thank you in advance...
Dr. Nejat Yilmaz
==============================Original Headers============================== 8, 28 -- From nyilmaz-at-mersin.edu.tr Fri Oct 13 14:17:22 2006 8, 28 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DJHLFG015879 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 14:17:22 -0500 8, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 8, 28 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 25FF94508A 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 22:32:59 +0300 (EEST) 8, 28 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr 8, 28 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 8, 28 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 8, 28 -- with ESMTP id K66xSD0aheDH for {Microscopy-at-microscopy.com} ; 8, 28 -- Fri, 13 Oct 2006 22:32:56 +0300 (EEST) 8, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 8, 28 -- by mail.mersin.edu.tr (Postfix) with SMTP id 21AA6450AE 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 22:32:56 +0300 (EEST) 8, 28 -- To: {Microscopy-at-microscopy.com} 8, 28 -- Subject: Skin processing 8, 28 -- From: {nyilmaz-at-mersin.edu.tr} 8, 28 -- Date: Fri, 13 Oct 2006 22:32:56 EEST 8, 28 -- Reply-To: {nyilmaz-at-mersin.edu.tr} 8, 28 -- Errors-To: {nyilmaz-at-mersin.edu.tr} 8, 28 -- X-Priority: 3 (Normal) 8, 28 -- X-Originating-Ip: [85.104.168.63] 8, 28 -- X-Mailer: NOCC v0.9.5 8, 28 -- Content-Type: text/plain; 8, 28 -- charset="ISO-8859-9" 8, 28 -- Content-Transfer-Encoding: 8bit 8, 28 -- Message-Id: {20061013193256.21AA6450AE-at-mail.mersin.edu.tr} ==============================End of - Headers==============================
==============================Original Headers============================== 18, 23 -- From lesley.bechtold-at-jax.org Fri Oct 13 14:36:20 2006 18, 23 -- Received: from carmen.jax.org (carmen.jax.org [192.43.249.16]) 18, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DJaKC4026581 18, 23 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 13 Oct 2006 14:36:20 -0500 18, 23 -- Received: from jcs-mid-prod.jax.org (jcs-mid-prod.jax.org [192.43.249.134]) 18, 23 -- by carmen.jax.org (8.12.11/8.12.11) with ESMTP id k9DIUA01026242; 18, 23 -- Fri, 13 Oct 2006 15:36:14 -0400 (EDT) 18, 23 -- (envelope-from lesley.bechtold-at-jax.org) 18, 23 -- Received: from spikey.jax.org by jcs-mid-prod.jax.org 18, 23 -- with ESMTP id 129656101160768136; Fri, 13 Oct 2006 15:35:36 -0400 18, 23 -- From: "Lesley Bechtold" {lesley.bechtold-at-jax.org} 18, 23 -- To: "nyilmaz-at-mersin.edu.tr" {nyilmaz-at-mersin.edu.tr} 18, 23 -- CC: "Microscopy Network" {microscopy-at-ns.microscopy.com} 18, 23 -- Subject: RE: [Microscopy] Skin processing 18, 23 -- Date: Fri, 13 Oct 2006 15:35:35 -0400 18, 23 -- Message-ID: {20061013153535960.00000006260-at-spikey} 18, 23 -- In-Reply-To: {200610131925.k9DJPOxc023765-at-ns.microscopy.com} 18, 23 -- X-Mailer: Oracle Connector for Outlook 10.1.1.0.5 71011 (11.0.8010) 18, 23 -- X-Accept-Language: en-us, en 18, 23 -- MIME-Version: 1.0 18, 23 -- Content-Type: text/plain; charset=us-ascii 18, 23 -- Content-Transfer-Encoding: 8bit 18, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9DJaKC4026581 ==============================End of - Headers==============================
Hi Nejat I do not have a protocol for TEM of skin, but Pierre Coulombe is the master. Contact him at coulombe-at-jhmi.edu. He can get you what you need. David
On Oct 13, 2006, at 12:21 PM, nyilmaz-at-mersin.edu.tr wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Dear Colleagues... } } We're trying to process skin tissue samples for TEM. Despite several } attempts with different protocols, we couldn't } achieve fully satisfactory results.Has anybody experienced skin } tissue processing? } Are there "tips and tricks" for } processing skin tissue? } Thank you in advance... } } Dr. Nejat Yilmaz } } } } } } } ==============================Original } Headers============================== } 8, 28 -- From nyilmaz-at-mersin.edu.tr Fri Oct 13 14:17:22 2006 } 8, 28 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr } [193.255.128.3]) } 8, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k9DJHLFG015879 } 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 } 14:17:22 -0500 } 8, 28 -- Received: from localhost (localhost.mersin.edu.tr } [127.0.0.1]) } 8, 28 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 25FF94508A } 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 } 22:32:59 +0300 (EEST) } 8, 28 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr } 8, 28 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) } 8, 28 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd- } new, port 10024) } 8, 28 -- with ESMTP id K66xSD0aheDH for {Microscopy-at-microscopy.com} ; } 8, 28 -- Fri, 13 Oct 2006 22:32:56 +0300 (EEST) } 8, 28 -- Received: from localhost (localhost.mersin.edu.tr } [127.0.0.1]) } 8, 28 -- by mail.mersin.edu.tr (Postfix) with SMTP id 21AA6450AE } 8, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 } 22:32:56 +0300 (EEST) } 8, 28 -- To: {Microscopy-at-microscopy.com} } 8, 28 -- Subject: Skin processing } 8, 28 -- From: {nyilmaz-at-mersin.edu.tr} } 8, 28 -- Date: Fri, 13 Oct 2006 22:32:56 EEST } 8, 28 -- Reply-To: {nyilmaz-at-mersin.edu.tr} } 8, 28 -- Errors-To: {nyilmaz-at-mersin.edu.tr} } 8, 28 -- X-Priority: 3 (Normal) } 8, 28 -- X-Originating-Ip: [85.104.168.63] } 8, 28 -- X-Mailer: NOCC v0.9.5 } 8, 28 -- Content-Type: text/plain; } 8, 28 -- charset="ISO-8859-9" } 8, 28 -- Content-Transfer-Encoding: 8bit } 8, 28 -- Message-Id: {20061013193256.21AA6450AE-at-mail.mersin.edu.tr} } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-arizona.edu Fri Oct 13 15:19:52 2006 5, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DKJo2b005573 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Oct 2006 15:19:51 -0500 5, 22 -- Received: from localhost (boromir.email.arizona.edu [10.0.0.217]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 8498911447A2 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Oct 2006 13:19:46 -0700 (MST) 5, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id D8574114B450 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 13 Oct 2006 13:19:37 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 22 -- In-Reply-To: {200610131921.k9DJLjng021171-at-ns.microscopy.com} 5, 22 -- References: {200610131921.k9DJLjng021171-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {2623A477-CC5A-4437-B692-39EA95A4157E-at-arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-arizona.edu} 5, 22 -- Subject: Re: [Microscopy] Skin processing 5, 22 -- Date: Fri, 13 Oct 2006 13:19:33 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.752.2) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
anybody out there believing /considering, that there are differences between human and animal tissue ??....if not, I could send my protocol for human, diagnostic skin tissue (embedding in epoxid resin...Glycidether100/DMSA/MNA/DMP-30)...the differences for embedding method perhaps are not that big......
best regards, Wolfgang Muss SALZBURG, Austria
---------- Von: nyilmaz-at-mersin.edu.tr[SMTP:nyilmaz-at-mersin.edu.tr] Antwort an: nyilmaz-at-mersin.edu.tr Gesendet: Samstag, 14. Oktober 2006 00:12 An: W.Muss-at-salk.at Betreff: [Microscopy] Addition for skin processing
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Dear colleagues...
We are using Spurrs resin or Araldit resin and we are working on rat skins.
Thanks again
Dr. Nejat Yilmaz
==============================Original Headers============================== 9, 28 -- From nyilmaz-at-mersin.edu.tr Fri Oct 13 17:07:02 2006 9, 28 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 9, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9DM71BN017947 9, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 13 Oct 2006 17:07:01 -0500 9, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 9, 28 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 1BEF145111 9, 28 -- for {Microscopy-at-microscopy.com} ; Sat, 14 Oct 2006 01:22:50 +0300 (EEST) 9, 28 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr 9, 28 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 9, 28 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 9, 28 -- with ESMTP id HEN7dc7jBGXE for {Microscopy-at-microscopy.com} ; 9, 28 -- Sat, 14 Oct 2006 01:22:47 +0300 (EEST) 9, 28 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 9, 28 -- by mail.mersin.edu.tr (Postfix) with SMTP id 1C0A84510B 9, 28 -- for {Microscopy-at-microscopy.com} ; Sat, 14 Oct 2006 01:22:47 +0300 (EEST) 9, 28 -- To: {Microscopy-at-microscopy.com} 9, 28 -- Subject: Addition for skin processing 9, 28 -- From: {nyilmaz-at-mersin.edu.tr} 9, 28 -- Date: Sat, 14 Oct 2006 01:22:47 EEST 9, 28 -- Reply-To: {nyilmaz-at-mersin.edu.tr} 9, 28 -- Errors-To: {nyilmaz-at-mersin.edu.tr} 9, 28 -- X-Priority: 3 (Normal) 9, 28 -- X-Originating-Ip: [85.104.168.63] 9, 28 -- X-Mailer: NOCC v0.9.5 9, 28 -- Content-Type: text/plain; 9, 28 -- charset="ISO-8859-9" 9, 28 -- Content-Transfer-Encoding: 8bit 9, 28 -- Message-Id: {20061013222247.1C0A84510B-at-mail.mersin.edu.tr} ==============================End of - Headers==============================
==============================Original Headers============================== 19, 39 -- From W.Muss-at-salk.at Sat Oct 14 06:16:03 2006 19, 39 -- Received: from hermes.salk.at (hermes.lks.at [193.170.167.9] (may be forged)) 19, 39 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9EBG2uW008724 19, 39 -- for {Microscopy-at-microscopy.com} ; Sat, 14 Oct 2006 06:16:03 -0500 19, 39 -- Received: from localhost (localhost [127.0.0.1]) 19, 39 -- by hermes.salk.at (Postfix) with ESMTP id D4B38C38A0; 19, 39 -- Sat, 14 Oct 2006 13:16:00 +0200 (CEST) 19, 39 -- X-Virii-Scanned: Kaspersky Antivirus at salk.at 19, 39 -- Received: from hermes.salk.at ([127.0.0.1]) 19, 39 -- by localhost (n1ex218.lks.local [127.0.0.1]) (amavisd-new, port 10024) 19, 39 -- with ESMTP id 3uecVCrpXpyB; Sat, 14 Oct 2006 13:16:00 +0200 (CEST) 19, 39 -- Received: from hermes.lks.at (hermes.salk.at [192.168.13.210]) 19, 39 -- by hermes.salk.at (Postfix) with ESMTP id 59134C3886; 19, 39 -- Sat, 14 Oct 2006 13:16:00 +0200 (CEST) 19, 39 -- Received: from localhost (localhost [127.0.0.1]) 19, 39 -- by hermes.lks.at (Postfix) with ESMTP id 36CAC5A9020; 19, 39 -- Sat, 14 Oct 2006 13:16:00 +0200 (CEST) 19, 39 -- Received: from hermes.lks.at ([127.0.0.1]) 19, 39 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 19, 39 -- with ESMTP id 23523-07; Sat, 14 Oct 2006 13:15:59 +0200 (CEST) 19, 39 -- Received: from c1pa003 (unknown [192.168.42.3]) 19, 39 -- by hermes.lks.at (Postfix) with SMTP id D9A715A900A; 19, 39 -- Sat, 14 Oct 2006 13:15:59 +0200 (CEST) 19, 39 -- Received: by localhost with Microsoft MAPI; Sat, 14 Oct 2006 13:15:59 +0200 19, 39 -- Message-ID: {01C6EF92.E40C1AA0.W.Muss-at-salk.at} 19, 39 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 19, 39 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 19, 39 -- To: "'nyilmaz-at-mersin.edu.tr'" {nyilmaz-at-mersin.edu.tr} 19, 39 -- Cc: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 19, 39 -- Subject: [Microscopy] RE: Addition for skin processing 19, 39 -- Date: Sat, 14 Oct 2006 13:15:58 +0200 19, 39 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 19, 39 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 19, 39 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 19, 39 -- MIME-Version: 1.0 19, 39 -- Content-Type: text/plain; charset="us-ascii" 19, 39 -- Content-Transfer-Encoding: 7bit 19, 39 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at 19, 39 -- X-Scanned-By: SALK-Content-Filter ==============================End of - Headers==============================
Thanks to all who replied and gave advice. I'll forward the info to our electrician. BTW, our CRT screen came on for 10 minutes this morning (Monday - the TEM is still relatively cold).
John Brealey
Quote... "I think we have the same scope of the same vintage. Likewise, over the past 2 yrs the CRT has had problems. Unusually long warm up times are the norm. It was about 10min...now it's up to 30 min before the CRT kicks in. After it warms up however..it has not been a problem at all. What I don't understand is how you operate blindly to set up numbers etc. It seems that if the CRT is not on, we cannot even get a beam. I've tried to run blind with it only a few times...maybe I'll have to do it again one day. I hope not. We are still under the Hitachi service contract, and it seems that every time they come over they try something new. They have swapped out the CRT and some electrical component boards. Nothing has worked thus far. I'll be interested in any real solutions that you can share as to using a stand alone monitor. Good luck. Linda"
Linda, we have the opposite problem to you - our CRT comes on when cold then disappears after about 10 minutes. I suspect we have a faulty wire somewhere or a fine crack on a circuit board that expands as it heats up. Despite the lack of illumination on the CRT screen, all functions performed through the CRT keypad are working. Your problem sounds different - maybe a power supply problem.
Quote... "You may use any composite video monitor (9" B&W security monitor), given that H-600 takes composite video- I am not 100% certain about H-600, but most data monitors like that use standard composite video 1V p-p; 75 Ohm. Worth checking out IMHO. Vitaly Feingold"
Thanks for the info Vitaly.
Quote... "Try to replace all the capacitors in the circuit board. It is possible the high voltage is not working for the crt."
Thanks Ray.
Quote... "It sounds like a heat related issue. I had to remember an old trick for a problem we recently had. I turned a duster can upside down and sprayed liquid around at some suspect components. When I sprayed the bad one, the unit snapped back to life momentarily. I could then replace the bad component on the first attempt. I would be careful around a CRT and wouldn't be chilling the glass lest something break, but I suspect something in the driver electronics has gone bad. My next recourse would be to look for a replacement CRT from an old scope. I've had to do that with two different EDS units. Warren"
Thanks Warren.
==============================Original Headers============================== 12, 33 -- From john.brealey-at-imvs.sa.gov.au Sun Oct 15 21:54:59 2006 12, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 12, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9G2swsr006822 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Sun, 15 Oct 2006 21:54:59 -0500 12, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 12, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k9G2svJZ005767 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 12:24:57 +0930 (CST)' 12, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 12, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k9G2svmJ005764 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 12:24:57 +0930 (CST)' 12, 33 -- Received: from localhost (scalix1.imvs.sa.gov.au [10.20.98.38]) 12, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id 25DD62EC28F 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 12:24:57 +0930 (CST) 12, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 12, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 12, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.38]) (amavisd-new, port 10024) 12, 33 -- with LMTP id 3fD+Ds9W6LTn for {Microscopy-at-MSA.Microscopy.Com} ; 12, 33 -- Mon, 16 Oct 2006 12:24:42 +0930 (CST) 12, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 12, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 6DC882EC294 12, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 12:24:42 +0930 (CST) 12, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 12, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 12, 33 -- Subject: Hitachi H-600 CRT Problem Summary 12, 33 -- Date: Mon, 16 Oct 2006 12:24:41 +0930 12, 33 -- Message-ID: {000601c6f0ce$6ded1120$c88a140a-at-iqe36042} 12, 33 -- MIME-Version: 1.0 12, 33 -- Content-Type: text/plain; 12, 33 -- charset="us-ascii" 12, 33 -- Content-Transfer-Encoding: 7bit 12, 33 -- X-Mailer: Microsoft Office Outlook 11 12, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 12, 33 -- Thread-Index: AcbwzlUpp5RNZK5yS5m/ESEwLpfxWw== ==============================End of - Headers==============================
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I am wondering what other multi-user centers use as benchmarks when justifying hiring additional staff. I can provide total hours of equipment usage and how many extra hours and weekends I am working but I am thinking there might be something more concrete I could use. Does any one have suggestions or gone through this recently?
Best Regards,
Kirk
Kirk J. Czymmek, Ph.D. Associate Professor Department of Biological Sciences Delaware Biotechnology Institute Bio-Imaging Center, Director University of Delaware Newark, DE 19711
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Email: annamaria.pisi Name: annamaria pisi
Organization: University
Title-Subject: [Filtered] TEM and SEM fixation
Question: I have to treat human stomach biopsies that had been frozen in liquid nitrogen and kept at -85ƒC without any protectant device. I have to fix and embedd some of them for TEM and some for SEM. What should I do? I never had samples like these? I have a traditional ultramicrotome and no cryo devices. Please help thank you annamaria
Annamaria, If these were simply plunged into liquid nitrogen to freeze them, the ultrastructure will be virtually destroyed by ice crystal growth. You might check how they were frozen before you proceed to see if it's worth it. That said you can freeze-substitute them then bring them to room temperature and embed as normal without cryodevices. Get a small Styrofoam cooler (the kind chemicals are shipped in) and some dry ice and you are in business.
A good substitution medium is 1% Osmium tetroxide in dry acetone (dissolve from the crystals). Add about 1 ml of substitution medium to 2 ml cryovials, cap the vials and freeze them upright in liquid nitrogen. Transfer your samples to the top of the frozen substitution medium in a liquid nitrogen bath and loosely cap the vials, making sure there is no liquid nitrogen inside. For the substitution chamber, line a small Styrofoam cooler with some foil then put 2 beakers in, one inside the other. Add acetone to each beaker to about 1/3 full then chill everything by surrounding the beakers with dry ice. Put a few chunks in the acetone, too. Once it's all cooled add your samples with substitution medium then top up the cooler with dry ice and put the lid on. Usually a small cooler holds about 5 pounds of dry ice which will last 2 to 5 days depending on the outside temperature, but this should be enough to complete the substitution. Once the samples warm up to room temperature they are already fixed, dehydrated and ready to embed in the resin of your choice (or to CPD and put in the SEM). Don't forget to rinse a few times in pure acetone to remove the excess Osmium.
Let me know if you need more information. Kim
annamaria.pisi-at-ns.microscopy.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both annamaria.pisi as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: annamaria.pisi } Name: annamaria pisi } } Organization: University } } Title-Subject: [Filtered] TEM and SEM fixation } } Question: I have to treat human stomach biopsies that had been frozen in liquid nitrogen and kept at -85ƒC without any protectant device. I have to fix and embedd some of them for TEM and some for SEM. What should I do? I never had samples like these? I have a traditional ultramicrotome and no cryo devices. Please help } thank you annamaria }
-- Kim Rensing Ph.D. Manager, Microscopy and Imaging Facility, University of Calgary, Health Sciences Centre B129 - 3330 Hospital Drive NW Calgary, AB, Canada T2N 4N1 O: 403-220-3488 F: 403-270-8928
==============================Original Headers============================== 6, 27 -- From krensing-at-ucalgary.ca Mon Oct 16 09:47:19 2006 6, 27 -- Received: from mr2.ucalgary.ca (mr2.ucalgary.ca [136.159.34.166]) 6, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GElIPa026874 6, 27 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 09:47:18 -0500 6, 27 -- Received: from smtp2.ucalgary.ca (smtp2.ucalgary.ca [136.159.34.223]) 6, 27 -- by mr2.ucalgary.ca (Postfix) with ESMTP id 4D115366A5; 6, 27 -- Mon, 16 Oct 2006 08:47:12 -0600 (MDT) 6, 27 -- Received: from [192.168.0.100] ([136.159.164.171]) 6, 27 -- (authenticated (0 bits)) 6, 27 -- by smtp2.ucalgary.ca (8.11.7/8.11.6) with ESMTP id k9GEl8013753 6, 27 -- (using TLSv1/SSLv3 with cipher DHE-RSA-AES256-SHA (256 bits) verified NO); 6, 27 -- Mon, 16 Oct 2006 08:47:08 -0600 6, 27 -- Message-ID: {45339B66.5070806-at-ucalgary.ca} 6, 27 -- Date: Mon, 16 Oct 2006 08:47:02 -0600 6, 27 -- From: Kim Rensing {krensing-at-ucalgary.ca} 6, 27 -- Organization: Microscopy and Imaging Facility, U. of Calgary 6, 27 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 6, 27 -- MIME-Version: 1.0 6, 27 -- To: annamaria.pisi-at-ns.microscopy.com, microscopy-at-microscopy.com 6, 27 -- Subject: Re: [Microscopy] viaWWW: TEM and SEM fixation human stomach 6, 27 -- References: {200610161420.k9GEKwOR025761-at-ns.microscopy.com} 6, 27 -- In-Reply-To: {200610161420.k9GEKwOR025761-at-ns.microscopy.com} 6, 27 -- Content-Type: text/plain; charset=windows-1252; format=flowed 6, 27 -- Content-Transfer-Encoding: 8bit 6, 27 -- X-UCalgary-MailScanner-Information: Please contact IT Help Desk at (403) 220-5555 for more information 6, 27 -- X-UCalgary-MailScanner: Found to be clean 6, 27 -- X-UCalgary-MailScanner-From: krensing-at-ucalgary.ca ==============================End of - Headers==============================
I am sure this is an easy one - I need to trace DNA (or RNA) molecules on a digital micrograph to measure their length. Any particular program recommended? NIH Image should do it? Or something maybe better (preferentially free/cheap)?
Thanks,
Michal
==============================Original Headers============================== 5, 17 -- From Michal.Jarnik-at-fccc.edu Mon Oct 16 10:15:06 2006 5, 17 -- Received: from azah.fccc.edu (azah.fccc.edu [131.249.4.237]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GFF6Qn005514 5, 17 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 10:15:06 -0500 5, 17 -- Received: from [131.249.6.85] (emf1.fccc.edu [131.249.6.85]) 5, 17 -- (authenticated bits=0) 5, 17 -- by azah.fccc.edu (8.13.6/8.13.6) with ESMTP id k9GFF5oP020664 5, 17 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 11:15:05 -0400 (EDT) 5, 17 -- Message-ID: {4533A1F3.90404-at-fccc.edu} 5, 17 -- Date: Mon, 16 Oct 2006 11:14:59 -0400 5, 17 -- From: Michal Jarnik {Michal.Jarnik-at-fccc.edu} 5, 17 -- User-Agent: Thunderbird 1.5.0.5 (Macintosh/20060719) 5, 17 -- MIME-Version: 1.0 5, 17 -- To: microscopy-at-microscopy.com 5, 17 -- Subject: Measuring length of DNA/RNA 5, 17 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 17 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I am posting this question for a friend who is working on yeast using the following (very odd) protocol:
"These yeast are chemically fixed in PB containing 3% paraformaldehyde and 0.5% gluteraldehyde 2 h room temp. Washed twice in PB, placed in 1% sodium meta periodate, rinsed PB, quenched for 30 min in 50 mM NH4Cl in PB, rinsed PB, dehydrated in ETOH 30%, 70%, 95%, and 3x 100%, microwaved 40s, 250W. Allowed to drain dry between each 100% ETOH. Placed in LR White and microwaved 3 min, 250W, placed 4 C overnight, with two more LR White changes, then placed in LE white in embedding capsules at 60 C overnight."
Go to http://www.emc.missouri.edu/lookatthis.htm to see these yeasty beasties.
She is looking for opinions on why the cell membranes are bumpy (I said drying out between 100% ETOH changes), why the vacuoles look so strange, and why the mottled appearance in the background (water in the formvar, she asks?). When reading the above protocol again, note that these samples are NOT destined for immunolabeling.
I can't wait to see the replies to this one!
Rainy day in Columbia. How's Hawaii, Tina?
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Mon Oct 16 13:01:51 2006 10, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GI1ohB019704 10, 23 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 13:01:51 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Mon, 16 Oct 2006 13:01:49 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="US-ASCII" 10, 23 -- Subject: Evil protocol? 10, 23 -- Date: Mon, 16 Oct 2006 13:01:49 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68AEE-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: Evil protocol? 10, 23 -- Thread-Index: AcbxTSdHxrhXluLORD+Z8CZucccncA== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 16 Oct 2006 18:01:49.0844 (UTC) FILETIME=[27296D40:01C6F14D] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9GI1ohB019704 ==============================End of - Headers==============================
1. I would recommend she use glutaraldehyde instead of gluteraldehyde :) 2. Why is she doing periodate? try skipping this step to see if it impacts the morphology. 3. Why NH4Cl if she isn't going to do immuno? 4. It is hard for me to judge a microwave dehydration protocol but try a conventional non-microwave approach. I would be worried about shrinkage. I agree the drying out step is highly suspect. 5. Why no osmium? 6. Why not use your fancy cryofixation device? - Studer has published gorgeous pictures of cryo-fixed yeast. 7. If no immuno, why not a better resin (epon-substitute or epon-araldite)? 8. Tina probably would need a battery operated computer to get your message!
At 01:03 PM 10/16/06, you wrote:
} ---------------------------------------------------------------------------- } 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
My weather comment to Tina was made while having a total blankout on the recent earthquake in Hawaii. It was definitely not meant as a bad joke at the expense of the people who went though it! My apologies if it came across like that. My bad all the way.
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 5, 23 -- From TindallR-at-missouri.edu Mon Oct 16 13:40:47 2006 5, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GIekmw009224 5, 23 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 13:40:47 -0500 5, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 5, 23 -- Mon, 16 Oct 2006 13:40:43 -0500 5, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 5, 23 -- Content-class: urn:content-classes:message 5, 23 -- MIME-Version: 1.0 5, 23 -- Content-Type: text/plain; 5, 23 -- charset="US-ASCII" 5, 23 -- Subject: Evil protocol oops 5, 23 -- Date: Mon, 16 Oct 2006 13:40:43 -0500 5, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68AF3-at-UM-XMAIL08.um.umsystem.edu} 5, 23 -- X-MS-Has-Attach: 5, 23 -- X-MS-TNEF-Correlator: 5, 23 -- Thread-Topic: Evil protocol oops 5, 23 -- Thread-Index: AcbxUpYLx8afvMr3S+mX6Hj+L8ADQA== 5, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 5, 23 -- To: {microscopy-at-microscopy.com} 5, 23 -- X-OriginalArrivalTime: 16 Oct 2006 18:40:43.0432 (UTC) FILETIME=[9616B280:01C6F152] 5, 23 -- Content-Transfer-Encoding: 8bit 5, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9GIekmw009224 ==============================End of - Headers==============================
Dear Listers, I would appreciate an advice on processing Drosophila mutant brain for TEM. I cannot use OsO4 or DMSO, and not sure which cryofixation and cryosubstitution approach would work the best. Thank you, Albina
-- MIKHAYLOVA,ALBINA, PhD
==============================Original Headers============================== 4, 21 -- From amich-at-ufl.edu Mon Oct 16 15:45:41 2006 4, 21 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GKjfTb022358 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 15:45:41 -0500 4, 21 -- Received: from osgjas03.cns.ufl.edu (osgjas03.cns.ufl.edu [128.227.74.133]) 4, 21 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k9GKjZLI5255366 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 16:45:35 -0400 4, 21 -- Message-ID: {49084854.20371161031535193.JavaMail.osg-at-osgjas03.cns.ufl.edu} 4, 21 -- Date: Mon, 16 Oct 2006 16:45:35 -0400 (EDT) 4, 21 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 4, 21 -- To: microscopy-at-microscopy.com 4, 21 -- Subject: Drosophila brain for TEM 4, 21 -- MIME-Version: 1.0 4, 21 -- Content-Type: text/plain; format=flowed; charset=us-ascii 4, 21 -- Content-Transfer-Encoding: 7bit 4, 21 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 4, 21 -- X-Originating-IP: 72.155.91.89 [72.155.91.89] 4, 21 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 21 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 21 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 21 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
1. The protocol is odd, if used for ultrastructure purpose. It is NOT odd at all if it is used for immunolabeling.
2. The "bumpy" cell membranes (looks to me the cell walls) may actually have something to do with the cell culture, rather than EM prep - some cells (the ones with bumpy cell walls) are mostly likely dying. It also explains, in part, the 'strangeness' of the vacuoles.
3. The mottled appearance in the background, if is from the formvar, could be easily checked out by examining the formvar on the grid (no sections).
4. The following is simple method for yeast TEM ultrastructure (especially if you want to avoid osmium). It works really well -
1). Harvested yeast cells are fixed first with 2.5% (v/v) glutaraldehyde in PBS for 40 min at room temperature. 2). Rinse with PBS 3X 3). Cells are further fixed with 2% potassium permanganate in water for 1 h at room temperature. 4). Rinse with water 5). Dehydration as usual 6). Use resin other than LRW for embedding
You can check some images at http://www.uwyo.edu/Microscopy/yatlas/default.htm
Zhaojie Zhang Director, Microscopy Core Facility University of Wyoming Laramie, WY 82072 http://www.uwyo.edu/microscopy
-----Original Message----- X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] Sent: Monday, October 16, 2006 11:12 AM To: Z.J. Zhang
Dear Listers,
I am posting this question for a friend who is working on yeast using the following (very odd) protocol:
"These yeast are chemically fixed in PB containing 3% paraformaldehyde and 0.5% gluteraldehyde 2 h room temp. Washed twice in PB, placed in 1% sodium meta periodate, rinsed PB, quenched for 30 min in 50 mM NH4Cl in PB, rinsed PB, dehydrated in ETOH 30%, 70%, 95%, and 3x 100%, microwaved 40s, 250W. Allowed to drain dry between each 100% ETOH. Placed in LR White and microwaved 3 min, 250W, placed 4 C overnight, with two more LR White changes, then placed in LE white in embedding capsules at 60 C overnight."
Go to http://www.emc.missouri.edu/lookatthis.htm to see these yeasty beasties.
She is looking for opinions on why the cell membranes are bumpy (I said drying out between 100% ETOH changes), why the vacuoles look so strange, and why the mottled appearance in the background (water in the formvar, she asks?). When reading the above protocol again, note that these samples are NOT destined for immunolabeling.
I can't wait to see the replies to this one!
Rainy day in Columbia. How's Hawaii, Tina?
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Mon Oct 16 13:01:51 2006 10, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GI1ohB019704 10, 23 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 13:01:51 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Mon, 16 Oct 2006 13:01:49 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="US-ASCII" 10, 23 -- Subject: Evil protocol? 10, 23 -- Date: Mon, 16 Oct 2006 13:01:49 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68AEE-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: Evil protocol? 10, 23 -- Thread-Index: AcbxTSdHxrhXluLORD+Z8CZucccncA== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 16 Oct 2006 18:01:49.0844 (UTC) FILETIME=[27296D40:01C6F14D] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9GI1ohB019704 ==============================End of - Headers==============================
==============================Original Headers============================== 26, 28 -- From ZZhang-at-uwyo.edu Mon Oct 16 17:52:22 2006 26, 28 -- Received: from ninemile.uwyo.edu (ninemile.uwyo.edu [129.72.10.29]) 26, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9GMqMJJ002908 26, 28 -- for {Microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 17:52:22 -0500 26, 28 -- Received: from UWMAIL.uwyo.edu (uwmail.uwyo.edu [172.26.4.76]) 26, 28 -- by ninemile.uwyo.edu (8.12.11.20060614/8.12.11) with ESMTP id k9GMqDI2003710; 26, 28 -- Mon, 16 Oct 2006 16:52:20 -0600 (MDT) 26, 28 -- (envelope-from ZZhang-at-uwyo.edu) 26, 28 -- Received: from TELEGRAPH5.uwyo.edu ([10.84.60.120]) by UWMAIL.uwyo.edu with Microsoft SMTPSVC(6.0.3790.211); 26, 28 -- Mon, 16 Oct 2006 16:52:05 -0600 26, 28 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 26, 28 -- Content-class: urn:content-classes:message 26, 28 -- MIME-Version: 1.0 26, 28 -- Content-Type: text/plain; 26, 28 -- charset="us-ascii" 26, 28 -- Subject: RE: [Microscopy] Evil protocol? 26, 28 -- Date: Mon, 16 Oct 2006 16:52:04 -0600 26, 28 -- Message-ID: {C9C1AF307F12AF4087DEF87CB0E6A4ADF65D33-at-TELEGRAPH5.uwyo.edu} 26, 28 -- In-Reply-To: {200610161811.k9GIBloX027956-at-ns.microscopy.com} 26, 28 -- X-MS-Has-Attach: 26, 28 -- X-MS-TNEF-Correlator: 26, 28 -- Thread-Topic: [Microscopy] Evil protocol? 26, 28 -- Thread-Index: AcbxTptO9VHFTH6mQDezomn3LaTRFAAEBQ8Q 26, 28 -- From: "Z.J. Zhang" {ZZhang-at-uwyo.edu} 26, 28 -- To: {TindallR-at-missouri.edu} , {Microscopy-at-microscopy.com} 26, 28 -- X-OriginalArrivalTime: 16 Oct 2006 22:52:05.0137 (UTC) FILETIME=[B37C4010:01C6F175] 26, 28 -- Content-Transfer-Encoding: 8bit 26, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9GMqMJJ002908 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both robert.clark-at-sharp.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: robert.clark-at-sharp.com Name: Rob
Organization: Sharp HealthCare
Title-Subject: [Filtered] TEM-Propylene Oxide v. Acetone
Question: Hi, What are the opions on using propylene oxide v. acetone mixed with resin in the final steps of resin infiltration of TEM processing? We use P.O. and have for years. The question came up as I am processing a piece of rubber tubing and am concerned the P.O. may eat away at it?
Used to be a service engineer for Hitachi now full time at U of Kentucky. I also refurbish electron microscopes for resale. I always loved the 600. I have been accumulating 600s for parts and probably have a crt around. I know I have the CRT I/O circuit board.
Couple thoughts on this if you are interested. Not certain on your problem but I suspect something with the CRT circuit unit board housed with the CRT itself. May be something with the CRT I/O board that is with all the other boards in the rack, lower left.
If you have access to a competent electronics guy especially with an oscilloscope you might try some things. On the CRT I/O board there are a couple test points; HD (horizontal drive) and VD (vertical drive). You can look at these. The schematics should tell you the frequencies. I doubt these are the problem however. Best place to look next is where the plug is to the board behind the CRT. Pain to remove the covers to get there. Take the grey large cover off, then the copper colored shroud off over the CRT. Need a long plus screwdriver for this. At the plug you will have HD pin 6, VD pin 9, Video pin 8, and +15 VDC pin 7. Pin 10 is ground. That suggestion of spraying duster upside down is a good idea once the trouble occurs. I would try and look at these when working fine and then again when it quits. I guess you have ten minutes. If you lose the +15 that will definitely cause this. If all these look OK you may be losing the +10KV to the CRT tube? Be careful around the plug and wire as it will wake you up. I would make sure the display power is off when messing with this plug. Even then the CRT will hold a charge. Once unplugged make sure and ground the tube. Usually a lot of carbon build up around the plug. Possible it is finding a way to ground. I would clean this area and the rubber cover. If your electronics guy has a high voltage probe and knows how not to get killed or short things out you should get ~8 to 10KV there when powered up. Don't think it will be right at 10KV if I remember right. Sometimes the wire to this pug and CRT may break down.
One clue to eliminate HD and/or VD is while running, visually inspect the plug end of the tube. Dark room you should some illumination on the grids in the glass.
Good luck. If I can help let me know.
Joel McClintock"
Thanks Joel, great info here. I'll forward this to our electrician - he's very good with oscilloscopes and stuff.
Quote...
"Hi John,
If you cant find a solution I would talk to these guys. SMH ELECTRONICS CO 508-291-7447 I had a monitor on an old Kevex unit that had a rolling line in it. They fixed it for a flat fee and it has been fine ever since. They specialize in component level repair of ancient electronics. They keep all the stuff from the 60's going for the FAA. They can probably diagnose and fix whatever you have.
Tom Kaye"
Thanks Tom.
==============================Original Headers============================== 17, 33 -- From john.brealey-at-imvs.sa.gov.au Mon Oct 16 19:07:01 2006 17, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 17, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9H06x5T003028 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 16 Oct 2006 19:07:00 -0500 17, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 17, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k9H06xJZ027770 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Oct 2006 09:36:59 +0930 (CST)' 17, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 17, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k9H06xmJ027767 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Oct 2006 09:36:59 +0930 (CST)' 17, 33 -- Received: from localhost (scalix1.imvs.sa.gov.au [10.20.98.38]) 17, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id E5927304501 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Oct 2006 09:36:58 +0930 (CST) 17, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 17, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 17, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.38]) (amavisd-new, port 10024) 17, 33 -- with LMTP id YL04FuXLCNih for {Microscopy-at-MSA.Microscopy.Com} ; 17, 33 -- Tue, 17 Oct 2006 09:36:41 +0930 (CST) 17, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 17, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id CF52C3044FE 17, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 17 Oct 2006 09:35:15 +0930 (CST) 17, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 17, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 17, 33 -- Subject: Hitachi H-600 CRT 17, 33 -- Date: Tue, 17 Oct 2006 09:35:14 +0930 17, 33 -- Message-ID: {000c01c6f17f$ec79be60$c88a140a-at-iqe36042} 17, 33 -- MIME-Version: 1.0 17, 33 -- Content-Type: text/plain; 17, 33 -- charset="us-ascii" 17, 33 -- Content-Transfer-Encoding: 7bit 17, 33 -- X-Mailer: Microsoft Office Outlook 11 17, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 17, 33 -- Thread-Index: Acbxf9OxNYQW5AZXRfGvJCmDebNFoQ== ==============================End of - Headers==============================
On 16 Oct 2006 at 19:00, bozhilov-at-ucr.edu wrote:
} } } } ---------------------------------------------------------------------- } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society } of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Can a photomultiplier operate directly in the vacuum of the specimen } chamber of a regular SEM, without being enclosed in a vacuum tube? } } } Krassimir Bozhilov } }
My guess, for the little that it may be worth, is that if the regular SEM used a diffusion pump, the PMT photocathode and the dynodes would soon become too oily to emit photoelectrons.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 7, 27 -- From r.sims-at-auckland.ac.nz Mon Oct 16 19:11:13 2006 7, 27 -- Received: from harpo.itss.auckland.ac.nz (mailhost.auckland.ac.nz [130.216.190.13]) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9H0BCCK012941 7, 27 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 19:11:13 -0500 7, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 7, 27 -- by harpo.itss.auckland.ac.nz (Postfix) with ESMTP id 21E7034A1B; 7, 27 -- Tue, 17 Oct 2006 13:11:12 +1300 (NZDT) 7, 27 -- Received: from harpo.itss.auckland.ac.nz ([127.0.0.1]) 7, 27 -- by localhost (smtpc.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 7, 27 -- with ESMTP id 03955-11; Tue, 17 Oct 2006 13:11:12 +1300 (NZDT) 7, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 7, 27 -- by harpo.itss.auckland.ac.nz (Postfix) with ESMTP id 020AB3453E; 7, 27 -- Tue, 17 Oct 2006 13:11:11 +1300 (NZDT) 7, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 7, 27 -- To: bozhilov-at-ucr.edu 7, 27 -- Date: Tue, 17 Oct 2006 13:15:56 +1300 7, 27 -- MIME-Version: 1.0 7, 27 -- Subject: Re: [Microscopy] PMT 7, 27 -- Cc: microscopy-at-microscopy.com 7, 27 -- Message-ID: {4534D78C.1885.11DAE85-at-localhost} 7, 27 -- Priority: normal 7, 27 -- In-reply-to: {200610170000.k9H007KA026517-at-ns.microscopy.com} 7, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 7, 27 -- Content-type: text/plain; charset=US-ASCII 7, 27 -- Content-transfer-encoding: 7BIT 7, 27 -- Content-description: Mail message body 7, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
A channel PMT might work in your situation. It eliminates the dynodes and voltage dropping resistors.
What type of application would you have in mind?
gary g.
At 05:00 PM 10/16/2006, you wrote:
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==============================Original Headers============================== 13, 20 -- From gary-at-gaugler.com Mon Oct 16 19:17:19 2006 13, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9H0HJG4024184 13, 20 -- for {microscopy-at-microscopy.com} ; Mon, 16 Oct 2006 19:17:19 -0500 13, 20 -- Received: (qmail 18999 invoked from network); 16 Oct 2006 17:17:18 -0700 13, 20 -- Received: by simscan 1.1.0 ppid: 18991, pid: 18993, t: 0.1493s 13, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 13, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 13, 20 -- by qsmtp2 with SMTP; 16 Oct 2006 17:17:18 -0700 13, 20 -- Message-Id: {7.0.1.0.2.20061016170953.02585ea8-at-gaugler.com} 13, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 20 -- Date: Mon, 16 Oct 2006 17:17:59 -0700 13, 20 -- To: bozhilov-at-ucr.edu 13, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 20 -- Subject: Re: [Microscopy] PMT 13, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 20 -- In-Reply-To: {200610170000.k9H00igN027809-at-ns.microscopy.com} 13, 20 -- References: {200610170000.k9H00igN027809-at-ns.microscopy.com} 13, 20 -- Mime-Version: 1.0 13, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I’m passing this on to the microscopy list with some comments.
I forgot to mention that we have a CM120 TWIN, which has a Cs of 2.2 mm, whereas your Biotwin has a Cs around 6 mm. That might explain the difference since Cs is one of the parameters that influence the envelope of the CTF.
On the other hand I don’t think an information limit of 20 Å is good enough even for a biotwin. My suggestion is to make the convergence angle as small as possible (sacrificing illumination intensity) by spreading the beam over a large specimen area. Maybe you have to use a larger spot to compensate for the loss of intensity.
I checked changing only Cs (between 2 and 6 mm) in CTF-explorer (http://clik.to/ctfexplorer) and it didn’t seem to affect the envelope very much, whereas the convergence angle certainly does.
I wonder if we’ll get any comments on this from the rest of the list.
Philip
Philip Koeck Karolinska Institute and Södertörns Högskola Dept. of Bioscience at Novum tel: +46 8 608 9186 fax: +46 8 608 9290 web: http://www.csb.ki.se/users/philip/philown.html ________________________________________ X-from: Pan Xijiang [mailto:panxijiang-at-gmail.com] Sent: 16 October 2006 18:16 To: Philip.Koeck
Hi Rob,
We have switched almost entirely to acetone for our dehydrating and infiltration steps. It works just fine with our variious permutations of Epon (EMBed), Spuur's, and Araldite and allows us to completely avoid the carcinogenic and hard-to-handle PO steps. We also use it occasionally as a transition between ethanol and resin in the rare cases when we want most of the dehydration done in the somewhat less-extractive ethanol. We use microwave processing for the vast majority of our samples, but it also works with longer procedures.
We have no experience with embedding rubber tubing, however.
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
-----Original Message----- X-from: robert.clark-at-sharp.com [mailto:robert.clark-at-sharp.com] Sent: Monday, October 16, 2006 6:48 PM To: Tindall, Randy D.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.msa.microscopy.org/MicroscopyListserver/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both robert.clark-at-sharp.com as well as the MIcroscopy Listserver ------------------------------------------------------------------------ ---
Email: robert.clark-at-sharp.com Name: Rob
Organization: Sharp HealthCare
Title-Subject: [Filtered] TEM-Propylene Oxide v. Acetone
Question: Hi, What are the opions on using propylene oxide v. acetone mixed with resin in the final steps of resin infiltration of TEM processing? We use P.O. and have for years. The question came up as I am processing a piece of rubber tubing and am concerned the P.O. may eat away at it?
Since you are asking it sounds like you have a problem. Having worked with a 100CX and 100S (23 yrs old and still going strong), the answer would have to be well in excess of 3000 plates for either one.
Two things seem to cause plate jams (in JEOL's): (1) plates loaded backwards in the film supply box (i.e. the notch on the wrong side, the boxes have just enough play in them that the weight of ~ 10 plate is enough to force a backwards plate at the bottom into the box. (2) bent plates.
Good luck.
On 11 Oct 2006 at 16:07, rcsencsits-at-lbl.gov wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Dear JEOL film camera users: } } What is your experience with the film camera? } How many successful film cassettes do you average between camera jams? } Please quote for each TEM model (100CX, 2000FX, 3100, etc). } } Thanks, } Roseann } } Roseann Csencsits, PhD. } Donner TEM Facility Manager } Lawrence Berkeley Lab } 510-486-4548 } } } } } ==============================Original Headers============================== } 7, 21 -- From rcsencsits-at-lbl.gov Wed Oct 11 16:06:48 2006 } 7, 21 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) } 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9BL6m4G028510 } 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 16:06:48 -0500 } 7, 21 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) } 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6laP008810 } 7, 21 -- for {Microscopy-at-Microscopy.Com} ; Wed, 11 Oct 2006 14:06:47 -0700 (PDT) } 7, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) } 7, 21 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id k9BL6kSY008805; } 7, 21 -- Wed, 11 Oct 2006 14:06:46 -0700 (PDT) } 7, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) } 7, 21 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } 7, 21 -- Message-Id: {DD626A78-3A2A-41AD-A7C9-81E173C72833-at-lbl.gov} } 7, 21 -- Content-Transfer-Encoding: 7bit } 7, 21 -- From: Roseann Csencsits {rcsencsits-at-lbl.gov} } 7, 21 -- Subject: JEOL camera question } 7, 21 -- Date: Wed, 11 Oct 2006 14:06:19 -0700 } 7, 21 -- To: Microscopy-at-Microscopy.Com } 7, 21 -- X-Mailer: Apple Mail (2.752.3) } 7, 21 -- X-Virus-Scanned: ClamAV 0.88.4/2025/Wed Oct 11 12:27:46 2006 on mta2 } 7, 21 -- X-Virus-Status: Clean } ==============================End of - Headers==============================
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 10, 27 -- From edelmare-at-muohio.edu Tue Oct 17 12:23:22 2006 10, 27 -- Received: from mulnx12.mcs.muohio.edu (mulnx12.mcs.muohio.edu [134.53.6.67]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9HHNMtk009258 10, 27 -- for {microscopy-at-Microscopy.com} ; Tue, 17 Oct 2006 12:23:22 -0500 10, 27 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 10, 27 -- by mulnx12.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9HHNJcO014117; 10, 27 -- Tue, 17 Oct 2006 13:23:20 -0400 10, 27 -- Received: from [192.168.1.23] ([134.53.14.97]) 10, 27 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9HHNI9X019832; 10, 27 -- Tue, 17 Oct 2006 13:23:19 -0400 10, 27 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 10, 27 -- To: rcsencsits-at-lbl.gov 10, 27 -- Date: Tue, 17 Oct 2006 13:23:31 -0400 10, 27 -- MIME-Version: 1.0 10, 27 -- Subject: Re: [Microscopy] JEOL camera question 10, 27 -- CC: microscopy-at-Microscopy.com 10, 27 -- Message-ID: {4534D953.29753.193DDC48-at-edelmare.muohio.edu} 10, 27 -- Priority: normal 10, 27 -- In-reply-to: {200610112107.k9BL7fRi029824-at-ns.microscopy.com} 10, 27 -- References: {200610112107.k9BL7fRi029824-at-ns.microscopy.com} 10, 27 -- X-mailer: Pegasus Mail for Windows (4.41) 10, 27 -- Content-type: text/plain; charset=US-ASCII 10, 27 -- Content-transfer-encoding: 7BIT 10, 27 -- Content-description: Mail message body 10, 27 -- X-Real-ConnectIP: 134.53.14.97 10, 27 -- X-Scanned-By: MIMEDefang 2.57 on 134.53.6.67 10, 27 -- X-Scanned-By: MIMEDefang 2.52 on 134.53.6.11 ==============================End of - Headers==============================
For a long time at CMU we tried to get a Digital System for our TEM. But out limitation was the lack of a 35mm port on the CM10. I didn't see that as a starting point for analysis.
#1 do you have a 35mm port? If so that's great. If not, open the wallet. As Mike mentioned the bottom mount camera systems typically are much more money.
(before more points another perspective)
Here in our facility we have a digital camera on our Philips 410. It has a 1k x 1k AMT (and no I have no financial interest in any vendors microscope or camera). The contrast and sensitivity means fast screening of images with intensities too low to visualize on the old 'analog' screen. The down side for us is the pixel size. Fewer pixels can mean increased sensitivity and decreased exposure times (faster live rates), but it means you are limited to on screen magnification. That has been the biggest challenge for me switching from film.
Film is great - take the image, and you have plenty of magnification overhead (enlarging). With the 1024x1024 camera, the image that you collect needs to be at the ultimate magnification. The sensitivity of the camera is such that the lower intensities work fine, but, goes counter the old adage of: take the image at the lowest magnification possible with the beam as far from cross over as possible.
With free software like autostich (google it) doing a montage is quite simple and direct (if you work with 8 bit files), but not quite the same as having a 2k or 4k array to work with. Even an automated montage system (assuming you have an automated stage that the camera software can integrate with) can overcome the limitations of few pixels, but potentially can add cost or exposure time, an still is not (in my limited opinion) quite as good as having the image collected on one array (film or CCD).
Last few thoughts to think about...
#2 How do you deal with the negatives: Routinely enlarge?
#3 Final print size (is a 1k x 1k image enough pixels)
#4 Speed and exposure times (goes to what Gary mentioned)
Good luck, and I hope that all makes sense (and I hope I'm not drastically repeating what others have already mentioned).
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: GBurgess-at-exchange.hsc.mb.ca [mailto:GBurgess-at-exchange.hsc.mb.ca] Sent: Thursday, October 12, 2006 3:50 PM To: Williams, Geoffrey
Dear TEM users, We are a pathology/hospital based TEM facility equipped with a Jeol 1010 electron microscope. We would like to change to a digital camera system, since we find that wet-film technology is slow and expensive. I am hoping that there are users out there that can provide me with opinions about digital camera systems to help educate me about what they like, and what they think is important in a TEM digital camera and its associated computer [hardware/software] system.
I have never used a TEM digital camera, so I have no experience as such with any digital camera system, other than consumer digital cameras, so it's hard for me to judge the merits of a digital camera without "getting my feet wet" so to speak, and trying it out.
I'm open to suggestions, thoughts, and opinions of experienced users or any comments on this topic.
Garry Burgess
Charge Technologist Electron Microscopy Department of Pathology Winnipeg, Canada
This e-mail and/or any documents in this transmission is intended for the address(s) only and may contain legally privileged or confidential information. Any unauthorized use, disclosure, distribution, copying or dissemination is strictly prohibited. If you receive this transmission in error, please notify the sender immediately and return the original.
==============================Original Headers============================== 10, 29 -- From GBurgess-at-exchange.hsc.mb.ca Thu Oct 12 14:41:35 2006 10, 29 -- Received: from hscxntmx0003.hsc.mb.ca (hscxntmx0003.hsc.mb.ca [142.233.100.122]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9CJfNDI031507 10, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 12 Oct 2006 14:41:29 -0500 10, 29 -- Received: from hscxmsmx0012.ad.wrha.mb.ca (unverified [172.16.6.178]) by 10, 29 -- hscxntmx0003.hsc.mb.ca(Vircom SMTPRS 4.0.346.0) with ESMTP id 10, 29 -- {B0025514934-at-hscxntmx0003.hsc.mb.ca} for {Microscopy-at-microscopy.com} ;Thu, 10, 29 -- 12 Oct 2006 14:41:44 -0500 10, 29 -- Received: from hscxmsmx0011.ad.wrha.mb.ca ([172.16.6.192]) by 10, 29 -- hscxmsmx0012.ad.wrha.mb.ca with Microsoft SMTPSVC(6.0.3790.1830); Thu, 12 10, 29 -- Oct 2006 14:41:22 -0500 10, 29 -- Message-ID: 10, 29 -- {CEC6924DC512E24FA19F6D142677B33665B981-at-hscxmsmx0011.ad.wrha.mb.ca} 10, 29 -- Date: Thu, 12 Oct 2006 14:41:13 -0500 10, 29 -- From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} 10, 29 -- Subject: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- To: {Microscopy-at-microscopy.com} 10, 29 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 29 -- Content-class: urn:content-classes:message 10, 29 -- X-MS-Has-Attach: 10, 29 -- X-MS-TNEF-Correlator: 10, 29 -- Thread-Topic: TEM Digital Camera - Opinions of Important Features Sought 10, 29 -- Thread-Index: AcbuNgj3H5J/RmFHRIuSUQI1vi57Cw== 10, 29 -- X-OriginalArrivalTime: 12 Oct 2006 19:41:22.0140 (UTC) 10, 29 -- FILETIME=[654679C0:01C6EE36] 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain 10, 29 -- Content-Transfer-Encoding: 8bit 10, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9CJfNDI031507 ==============================End of - Headers==============================
==============================Original Headers============================== 29, 30 -- From Geoffrey_Williams-at-brown.edu Tue Oct 17 14:43:44 2006 29, 30 -- Received: from draco.services.brown.edu (draco.services.brown.edu [128.148.106.172]) 29, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9HJhiR6022478 29, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 14:43:44 -0500 29, 30 -- Received: from ex-gateway2-out.AD.Brown.Edu (ex-gateway2.ad.brown.edu [128.148.21.53]) 29, 30 -- by draco.services.brown.edu (Switch-3.1.10/Switch-3.1.7/) with ESMTP id k9HJhhiR015046 29, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 15:43:43 -0400 (EDT) 29, 30 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway2-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 29, 30 -- Tue, 17 Oct 2006 15:43:43 -0400 29, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 29, 30 -- Content-class: urn:content-classes:message 29, 30 -- MIME-Version: 1.0 29, 30 -- Content-Type: text/plain; 29, 30 -- charset="us-ascii" 29, 30 -- Subject: RE: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 29, 30 -- Date: Tue, 17 Oct 2006 15:43:42 -0400 29, 30 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F04ACAF61-at-MAIL1.AD.Brown.Edu} 29, 30 -- In-Reply-To: {200610121950.k9CJoHMl005823-at-ns.microscopy.com} 29, 30 -- X-MS-Has-Attach: 29, 30 -- X-MS-TNEF-Correlator: 29, 30 -- Thread-Topic: [Microscopy] TEM Digital Camera - Opinions of Important Features Sought 29, 30 -- Thread-Index: AcbuN6X4PVQoLM5RQP25bN9/NOU+/gD6knpA 29, 30 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 29, 30 -- To: {Microscopy-at-microscopy.com} 29, 30 -- X-OriginalArrivalTime: 17 Oct 2006 19:43:43.0588 (UTC) FILETIME=[8DA67A40:01C6F224] 29, 30 -- X-Brown-Proofpoint: Not Infected 29, 30 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0609290000 definitions=main-0610170010 29, 30 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0609290000 definitions=main-0610170010 29, 30 -- Content-Transfer-Encoding: 8bit 29, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9HJhiR6022478 ==============================End of - Headers==============================
Our lab is disposing of a JEOL 2000FX TEM/STEM (see specs below). Please contact me off-line if you are interested in this instrument.
Specifications for JEOL 2000FX TEM Model # JEOL 2000FX TEM/STEM Serial # EM133042-32
Capabilities and features Purchased in 1987 Excellent condition; maintained by JEOL personnel; no serious outstanding or recurring problems 40 kV to 200 kV LaB6 gun Diffusion and ion pumped column and camera Specimen goniometer with 60 travel STEM capability using Polaroid film or 35-mm image acquisition Cold finger for sample EDS compatible TEM is not RS232 capable Air compressor to run pneumatic valves is not available (runs on house nitrogen in our facility) Gatan digital image acquisition system (see below) Equipment for plate camera (see below) Haskris R100 water-cooled chiller (1 year old; uses R134a coolant) Stand-alone film desiccator (without mechanical pump) JEOL speciality tools, includes hoist for gun assembly Documentation
Cameras and detectors: Gatan MSC-794 digital camera, 1024 x 1024 pixel CCD, lower mount (below plate camera); computer, monitor and software not included Polaroid film/CRT imaging system for STEM Plate camera for glass or film plates (3.25 x 4†plates) with 60 plate carriers and two sets of sender/receiver boxes 35-mm camera (for mounting on side port) STEM detectors Secondary Back-scattered Bright-field Annular dark-field
Specimen holders Standard two-grid holder EM-STH10: two-axis rotating holder EM-SCSH10: low-background holder EM-SRH10: specimen tilt-rotation holder EM-SCH: cryogenic holder and controller Gatan heating holder and controller
Gary M. Brown Microscopy Specialist ExxonMobil Chemical Company Baytown Technology & Engineering - West 5200 Bayway Drive Baytown, Texas 77520-2101 phone: 281 834 2387 fax: 281 834 2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
==============================Original Headers============================== 8, 20 -- From gary.m.brown-at-exxonmobil.com Tue Oct 17 14:44:57 2006 8, 20 -- Received: from hoespc01.exxonmobil.com (hoespc01.exxonmobil.com [192.67.48.38]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9HJivTp023383 8, 20 -- for {microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 14:44:57 -0500 8, 20 -- Received: from hounmg03.NA.XOM.COM (hounmg03.na.xom.com [158.35.101.41]) 8, 20 -- by hoespc01.exxonmobil.com (Switch-3.1.10/Switch-3.1.7) with ESMTP id k9HJiutp002521 8, 20 -- for {microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 14:44:56 -0500 (CDT) 8, 20 -- Subject: Disposal of JEOL 2000FX TEM 8, 20 -- Importance: 8, 20 -- To: microscopy-at-microscopy.com 8, 20 -- X-Mailer: Lotus Notes 652HF109 January 14, 2005 8, 20 -- Message-ID: {OFBA38CA8C.4AEE5F37-ON8625720A.006BCD58-8625720A.006C7B13-at-exxonmobil.com} 8, 20 -- From: gary.m.brown-at-exxonmobil.com 8, 20 -- Date: Tue, 17 Oct 2006 14:44:52 -0500 8, 20 -- X-MIMETrack: Serialize by Router on Hounmg03.na.xom.com/S/ExxonMobil(652FP1HF193|March 8, 20 -- 02, 2006) at 10/17/2006 02:44:55 PM 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-type: text/plain; charset=UTF-8 8, 20 -- Content-Transfer-Encoding: 8bit 8, 20 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id k9HJivTp023383 ==============================End of - Headers==============================
1) low energy secondary electrons collection will require an additional electrode at some +300V, same as standard SED has;
2) poor vacuum of specimen chamber will likely poison high resistance layer of the channel electron multiplier. These require much better vacuum.
Vitaly Feingold SIA 2773 Heath Line Duluth GA 30096 Ph. 770-232-7785 Fax 770-232-1791 www.sia-cam.com ----- Original Message ----- X-from: {gary-at-gaugler.com} To: {vitalylazar-at-att.net} Sent: Monday, October 16, 2006 8:18 PM
==============================Original Headers============================== 7, 23 -- From vitalylazar-at-att.net Tue Oct 17 14:50:19 2006 7, 23 -- Received: from mtiwmhc11.worldnet.att.net (mtiwmhc11.worldnet.att.net [204.127.131.115]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9HJoJkJ003771 7, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 17 Oct 2006 14:50:19 -0500 7, 23 -- Received: from siap6segs20pa8 (burnham.ljcrf.edu[192.231.106.2]) 7, 23 -- by worldnet.att.net (mtiwmhc11) with SMTP 7, 23 -- id {20061017195018111009m86ue} ; Tue, 17 Oct 2006 19:50:18 +0000 7, 23 -- Message-ID: {005b01c6f225$755711b0$0cf910ac-at-siap6segs20pa8} 7, 23 -- From: "Vitaly Feingold" {vitalylazar-at-att.net} 7, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 7, 23 -- References: {200610170018.k9H0IDkr026649-at-ns.microscopy.com} 7, 23 -- Subject: Re: [Microscopy] Re: PMT 7, 23 -- Date: Tue, 17 Oct 2006 15:50:07 -0400 7, 23 -- MIME-Version: 1.0 7, 23 -- Content-Type: text/plain; 7, 23 -- format=flowed; 7, 23 -- charset="iso-8859-1"; 7, 23 -- reply-type=original 7, 23 -- Content-Transfer-Encoding: 7bit 7, 23 -- X-Priority: 3 7, 23 -- X-MSMail-Priority: Normal 7, 23 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 7, 23 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
Actually you may simply skip this step. You can do the infiltration steps with a mixture of resin and 100% ethanol.
Regards,
Stephane
----- Original Message ---- X-from: "robert.clark-at-sharp.com" {robert.clark-at-sharp.com} To: nizets2-at-yahoo.com Sent: Tuesday, October 17, 2006 1:51:48 AM
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Email: robert.clark-at-sharp.com Name: Rob
Organization: Sharp HealthCare
Title-Subject: [Filtered] TEM-Propylene Oxide v. Acetone
Question: Hi, What are the opions on using propylene oxide v. acetone mixed with resin in the final steps of resin infiltration of TEM processing? We use P.O. and have for years. The question came up as I am processing a piece of rubber tubing and am concerned the P.O. may eat away at it?
I would be careful about this - I have found that traces of alcohol can adversely affect polymerisation. I do have to use ethanol/resin mix when processing cells that have been cultured on plastic dishes, but always give extra changes in pure resin in a vacuum incubator to ensure removal of trace ethanol.
For info - it is our normal practice to use acetone as a link reagent, having changed from using PO some years ago.
Alastair
At 04:12 18/10/2006 -0500, you wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Alastair McKinnon Histology & EM Facility Manager University of Aberdeen, Institute of Medical Science Foresterhill, Aberdeen, AB25 2ZD tel: +44(0) 1224 552923 (office); +44(0) 1224 555911 (lab) fax: +44(0) 1224 559533; email: a.d.mckinnon-at-abdn.ac.uk www.abdn.ac.uk/ims/h-em
==============================Original Headers============================== 11, 18 -- From a.d.mckinnon-at-abdn.ac.uk Wed Oct 18 05:07:14 2006 11, 18 -- Received: from mailhub1.abdn.ac.uk (mailhub1.abdn.ac.uk [139.133.7.28]) 11, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IA7DgV015183 11, 18 -- for {Microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 05:07:13 -0500 11, 18 -- Received: from med-0069.ims.abdn.ac.uk ([139.133.159.90] helo=med-0069.abdn.ac.uk) 11, 18 -- by mailhub1.abdn.ac.uk with esmtp (Exim 4.52) 11, 18 -- id 1Ga8KO-0003JN-En; Wed, 18 Oct 2006 11:07:12 +0100 11, 18 -- Message-Id: {5.2.1.1.0.20061018105834.01137098-at-mailms.abdn.ac.uk} 11, 18 -- X-Sender: pat081-at-mailms.abdn.ac.uk 11, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 11, 18 -- Date: Wed, 18 Oct 2006 11:07:10 +0100 11, 18 -- To: nizets2-at-yahoo.com 11, 18 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 11, 18 -- Subject: Re: [Microscopy] Re: viaWWW: TEM-Propylene Oxide v. Acetone 11, 18 -- Cc: Microscopy-at-microscopy.com 11, 18 -- In-Reply-To: {200610180912.k9I9CHJa012381-at-ns.microscopy.com} 11, 18 -- Mime-Version: 1.0 11, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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Email: hx213-at-cam.ac.uk Name: Huixin Xiu
Organization: University of Cambridge
Title-Subject: [Filtered] About the defect contrast in HAADF image
Question: Dear Microscopists,
I observed a strong defect ( possibly inversion domains) contrast in STEM-HAADF image at edge-on condition in GaN. The contrast also exists along {11-20} zone axis, but it is weaker than edge-on condition. I tried to use EDX to see whether any chemical change across the defect, but didn't find obvious change. Could anybody have similar observations or have any idea of the contrast from? Thanks a lot.
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 rfklie-at-uic.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rfklie-at-uic.edu Name: Robert Klie
Organization: University of Illinois - Chicago
Title-Subject: [Filtered] Postdoctoral Position available
Question: Dear all,
I am currently looking for a postdoctoral research associate in the area of STEM/EELS of oxide materials at the University of Illinois - Chicago. Would you please bring the advertisement (see below) to the attention of possible candidates?
Thanks!
Robert Klie -----------------------------------
POSTDOCTORAL POSITION AVAILABLE STARTING JANUARY 2007 DEPARTMENT OF PHYSICS UNIVERSITY OF ILLINOIS - CHICAGO, IL, USA
The University of Illinois ñ Chicago (UIC), Department of Physics, is seeking candidates for a postdoctoral position in transmission electron microscopy of strongly correlated oxides and heterogeneous catalysts systems. Applicants should have a Ph.D. degree in Materials Physics or Materials Chemistry, extensive and demonstrated experience in Z-contrast imaging in combination with electron energy-loss spectroscopy (EELS), and a strong background and interest in materials problem solving. Familiarity with aberration correction and analysis of structural defects and interfaces using electron-microscopy imaging is an advantage.
Facilities at UIC include a JEOL 2010F STEM/TEM, an aberration-corrected VG HB601UX, both with HAADF detectors and EELS spectrometers, a JEOL3010, as well as state-of-the-art sample preparation (for more details see http://www.rrc.uic.edu/).
The position is available starting January, 2007 for a period one year with a possible extension for a second year. The salary is commensurate with experience. Interested candidates should send a curriculum vitae, publication list, and the names of at least three references with their contact addresses to: Professor Robert F. Klie at rfklie-at-uic.edu
-------------------------------------------------------------------------------- Robert F. Klie, PhD Center for Functional Nanomaterials (Bldg.480) Brookhaven National Laboratory Upton, NY 11973 Tel. (631) 344-7709 Fax. (631)344-4071
AND
Assistant Professor University of Illinois at Chicago Department of Physics Chicago, IL 60607
There is a postdoctoral vacancy at the Universidad Técnica Federico Santa María. This is in the Vina del Mar/Valparaiso region of Chile. It is a good university and a great place to live. I recommend it for your consideration. Details below.
Alwyn Eades
Universidad Técnica Federico Santa María Condensed Mater Physics Group
3-years Postdoc position on Nanoscale Materials
Programa Bicentenario de Ciencia y Tecnología.
Our Condensed Mater Physics group at USM is focusing its research in both theoretical and experimental nanoscale materials. The current topics of research are: CVD synthesis of carbon nanotubes and other carbon nanostructures, synthesis of nanoparticle-nanotube hybrids, STM- AFM studies on the growth of metal thin films on crystalline surfaces in systems such as Ag/Al(100), Fe/CuAl(100), Fe/ZnO(100); Inverse Photoemission Spectroscopy of thin films and nanostructures; Low energy ion scattering studies of nanostructured materials; Theoretical studies of electronic structure of nanostructures under external electromagnetic fields.
We have one post-doc position open for somebody who fits with our research motivations. We are looking for a dynamic individual with expertise in either theory or experimental areas.
Essential requirements for candidates are: 1) A PhD in experimental or theoretical Physics, Chemistry or Materials Science. 2) A track record of quality publications 3) For experimentalists, experience with UHV and electron spectroscopies is desirable but not necessary. 4) For theorists, previous work on nanoscale systems and materials is required. 5) Excellent writing and communication skills are desired.
The position is initially opened for one year, and available immediately. Renewal up to 3-years upon mutual agreement.
Please send your CV and list of publications to Dr. Patricio Häberle (patricio.haberle-at-usm.cl)
-- ........... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
==============================Original Headers============================== 13, 24 -- From jae5-at-lehigh.edu Wed Oct 18 09:30:09 2006 13, 24 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IEU97O019280 13, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 09:30:09 -0500 13, 24 -- Received: from [127.0.0.1] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 13, 24 -- (authenticated bits=0) 13, 24 -- by rain.CC.Lehigh.EDU (8.13.8/8.13.8) with ESMTP id k9IEU73K028265 13, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 13, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 10:30:07 -0400 13, 24 -- Message-ID: {45363A6F.8000904-at-lehigh.edu} 13, 24 -- Date: Wed, 18 Oct 2006 10:30:07 -0400 13, 24 -- From: Alwyn Eades {jae5-at-lehigh.edu} 13, 24 -- Organization: Lehigh University 13, 24 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 13, 24 -- MIME-Version: 1.0 13, 24 -- To: MSA listserver {Microscopy-at-MSA.Microscopy.Com} 13, 24 -- Subject: Postdoctoral vacancy 13, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 13, 24 -- Content-Transfer-Encoding: 8bit 13, 24 -- X-Virus-Scanned: ClamAV version 0.88.5, clamav-milter version 0.88.5 on rain.CC.Lehigh.EDU 13, 24 -- X-Virus-Status: Clean 13, 24 -- X-Spam-Status: No, score=-1.4 required=5.0 tests=ALL_TRUSTED 13, 24 -- autolearn=disabled version=3.1.7 13, 24 -- X-Spam-Checker-Version: SpamAssassin 3.1.7 (2006-10-05) on rain.CC.Lehigh.EDU ==============================End of - Headers==============================
Today, I shot some images of histological sections in a conventional light microscope (Leica Orthoplan 2) using several different digital camera systems (Pixera, Qimaging). None of the images were uniformly in focus (i.e., they were not flat field) even though the microscope image itself was.
Is this a common problem with digital camera optical couplers? If anyone has a system that would work with this microscope setup, I would like to hear about it.
Frustrated in Carbondale,
J Bozzola -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
==============================Original Headers============================== 4, 18 -- From bozzola-at-siu.edu Wed Oct 18 15:40:25 2006 4, 18 -- Received: from abbmta1.siu.edu (abbmta1.siu.edu [131.230.254.205]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IKeL3d004713 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 18 Oct 2006 15:40:23 -0500 4, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 4, 18 -- by abbmta1.siu.edu (Switch-3.1.10/Switch-3.1.10) with ESMTP id k9IKeFuL021132 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 18 Oct 2006 15:40:17 -0500 (CDT) 4, 18 -- Mime-Version: 1.0 4, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 4, 18 -- Message-Id: {p06110427c15c3dccf719-at-[131.230.177.142]} 4, 18 -- Date: Wed, 18 Oct 2006 15:40:27 -0500 4, 18 -- To: Microscopy-at-msa.microscopy.com 4, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 4, 18 -- Subject: LM: digital camera lacking flat field 4, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 18 -- X-Spam-Score: 0.00% 4, 18 -- X-MASF: 0.00% 4, 18 -- X-Whitelist: 0.00% ==============================End of - Headers==============================
The sharpness of EBSD patterns decreases as the SEM kV is reduced.
Does anybody know why?
The only explanation I can come up with is stray fields. The lower kV electrons are going to be more affected by stray fields.
Regards, -- Larry Stoter
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==============================Original Headers============================== 5, 14 -- From larry-at-cymru.freewire.co.uk Wed Oct 18 15:46:12 2006 5, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 5, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IKkBWi009630 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 15:46:12 -0500 5, 14 -- Received: from [217.154.251.81] (th6dc-217-154-251-81.dial.mistral.co.uk [217.154.251.81] (may be forged)) 5, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9IKk9DI027032 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 21:46:10 +0100 5, 14 -- Mime-Version: 1.0 5, 14 -- Message-Id: {p06210200c15c4186a7b4-at-[217.154.250.196]} 5, 14 -- Date: Wed, 18 Oct 2006 21:43:02 +0100 5, 14 -- To: Microscopy-at-MSA.Microscopy.Com 5, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 5, 14 -- Subject: EBSD - kV dependence of pattern sharpness 5, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Dear Larry, I would suspect that either you are getting more contribution from the amorphous surface oxide layer as the beam penetrates less at lower kV, or that you are getting more contribution from the thin, deformed layer left over from polishing. The EBSP electrons come from very shallow in the specimen, because the ones from deeper in the sample don't come out with enough energy to contribute to the pattern, so the pattern is very sensitive to the condition of the surface and top micron of the sample. IMHO. Regards,
Mary Mager Electron Microscopist Materials Eng. UBC #419 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA Phone: 604-822-5648 Fax: 604-822-3619 email: mager-at-interchange.ubc.ca -----Original Message----- X-from: larry-at-cymru.freewire.co.uk [mailto:larry-at-cymru.freewire.co.uk] Sent: Wednesday, October 18, 2006 1:51 PM To: mager-at-interchange.ubc.ca
The sharpness of EBSD patterns decreases as the SEM kV is reduced.
Does anybody know why?
The only explanation I can come up with is stray fields. The lower kV electrons are going to be more affected by stray fields.
Regards, -- Larry Stoter
PLEASE NOTE - Agressive SPAM filtering is employed. If you don't get a reply to e-mails, try again, avoiding anything in the subject or body which might trigger filtering. If you are in the address book, you should get through. If you aren't, then there's a chance your e-mail will never be seen.
==============================Original Headers============================== 5, 14 -- From larry-at-cymru.freewire.co.uk Wed Oct 18 15:46:12 2006 5, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 5, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IKkBWi009630 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 15:46:12 -0500 5, 14 -- Received: from [217.154.251.81] (th6dc-217-154-251-81.dial.mistral.co.uk [217.154.251.81] (may be forged)) 5, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9IKk9DI027032 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 21:46:10 +0100 5, 14 -- Mime-Version: 1.0 5, 14 -- Message-Id: {p06210200c15c4186a7b4-at-[217.154.250.196]} 5, 14 -- Date: Wed, 18 Oct 2006 21:43:02 +0100 5, 14 -- To: Microscopy-at-MSA.Microscopy.Com 5, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 5, 14 -- Subject: EBSD - kV dependence of pattern sharpness 5, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 13, 33 -- From mager-at-interchange.ubc.ca Wed Oct 18 16:19:11 2006 13, 33 -- Received: from mta6.mail-relay.ubc.ca (mta6.mail-relay.ubc.ca [137.82.45.12]) 13, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9ILJBre026521 13, 33 -- for {microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 16:19:11 -0500 13, 33 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 13, 33 -- by mta6.mail-relay.ubc.ca (8.12.11.20060308/8.12.11) with ESMTP id k9ILJ4CV024400 13, 33 -- for {microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 14:19:04 -0700 (PDT) 13, 33 -- (envelope-from mager-at-interchange.ubc.ca) 13, 33 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 13, 33 -- by smtp.interchange.ubc.ca 13, 33 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 13, 33 -- with ESMTPA id {0J7C00JM8OJSC4-at-smtp.interchange.ubc.ca} for 13, 33 -- microscopy-at-microscopy.com; Wed, 18 Oct 2006 14:19:04 -0700 (PDT) 13, 33 -- Date: Wed, 18 Oct 2006 14:19:34 -0700 13, 33 -- From: Mary Mager {mager-at-interchange.ubc.ca} 13, 33 -- Subject: RE: [Microscopy] EBSD - kV dependence of pattern sharpness 13, 33 -- In-reply-to: {200610182050.k9IKokZ5023599-at-ns.microscopy.com} 13, 33 -- To: larry-at-cymru.freewire.co.uk 13, 33 -- Cc: Microscopy {microscopy-at-microscopy.com} 13, 33 -- Reply-to: mager-at-interchange.ubc.ca 13, 33 -- Message-id: {0J7C00JM9OJSC4-at-smtp.interchange.ubc.ca} 13, 33 -- Organization: Materials Eng. UBC 13, 33 -- MIME-version: 1.0 13, 33 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1478 13, 33 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 13, 33 -- Content-type: text/plain; charset=us-ascii 13, 33 -- Content-transfer-encoding: 7bit 13, 33 -- Thread-index: Acby9xbMWkhIiYRETgmPcFnvh3tPMQAAkPvg 13, 33 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.10.18.135442 13, 33 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 13, 33 -- X-PerlMx-Spam: Probability=7%, Report=__C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __USER_AGENT_MS_GENERIC 0 13, 33 -- X-Spam-Level: 13, 33 -- X-Spam-Flag: No ==============================End of - Headers==============================
Need some info on the fluid used in the water chilled lens cooling loop of a Jeol 733.
System developed a leak and dumped what at first looked like pump oil on the floor of the lab. After some clean up the fluid evaporated over the next few days which led me to think it was some kind of water based liquid.
Any 733 owners with info please contact me directly.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-smu.edu
==============================Original Headers============================== 9, 23 -- From rbeavers-at-mail.smu.edu Wed Oct 18 16:41:20 2006 9, 23 -- Received: from s31xub.systems.smu.edu (s31xub.systems.smu.edu [129.119.70.73]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9ILfJGL004997 9, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- Received: from s31xe7.systems.smu.edu ([129.119.70.140]) by s31xub.systems.smu.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 -- Content-Type: text/plain; 9, 23 -- charset="us-ascii" 9, 23 -- Subject: Jeol 733 Microprobe 9, 23 -- Date: Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- Message-ID: {F8AF6BF6CD1CC040AF35B0C2D1680BBB235E4A-at-s31xe7.systems.smu.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Jeol 733 Microprobe 9, 23 -- Thread-Index: Acby/iWDxPLNpHquTV69e0YFSFCLwA== 9, 23 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} 9, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 18 Oct 2006 21:41:19.0350 (UTC) FILETIME=[25A01160:01C6F2FE] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9ILfJGL004997 ==============================End of - Headers==============================
Roy, I've not done a whole lot of work on the JXA-733, but the objective lens is definitely OIL cooled. Don't go putting water in there! I'm just not sure what kind of oil. Maybe someone out there can help out with this.
Ken Converse owner
QUALITY IMAGES Servicing Scanning Electron Microscopes Since 1981 474 So. Bridgton Rd. Bridgton, ME 04009 207-647-4348 Fax 207-647-2688 kenconverse-at-qualityimages.biz qualityimages.biz
-----Original Message----- X-from: rbeavers-at-mail.smu.edu [mailto:rbeavers-at-mail.smu.edu] Sent: Wednesday, October 18, 2006 5:44 PM To: kenconverse-at-qualityimages.biz
Group,
Need some info on the fluid used in the water chilled lens cooling loop of a Jeol 733.
System developed a leak and dumped what at first looked like pump oil on the floor of the lab. After some clean up the fluid evaporated over the next few days which led me to think it was some kind of water based liquid.
Any 733 owners with info please contact me directly.
Thanks
Roy Beavers
Southern Methodist University Department of Geological Sciences P.O. Box 750395 Dallas, TX 75275 Voice: 214-768-2756 Fax: 214-768-2701 Email: rbeavers-at-smu.edu
==============================Original Headers============================== 9, 23 -- From rbeavers-at-mail.smu.edu Wed Oct 18 16:41:20 2006 9, 23 -- Received: from s31xub.systems.smu.edu (s31xub.systems.smu.edu [129.119.70.73]) 9, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9ILfJGL004997 9, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- Received: from s31xe7.systems.smu.edu ([129.119.70.140]) by s31xub.systems.smu.edu with Microsoft SMTPSVC(6.0.3790.1830); 9, 23 -- Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 23 -- Content-class: urn:content-classes:message 9, 23 -- MIME-Version: 1.0 9, 23 -- Content-Type: text/plain; 9, 23 -- charset="us-ascii" 9, 23 -- Subject: Jeol 733 Microprobe 9, 23 -- Date: Wed, 18 Oct 2006 16:41:19 -0500 9, 23 -- Message-ID: {F8AF6BF6CD1CC040AF35B0C2D1680BBB235E4A-at-s31xe7.systems.smu.edu} 9, 23 -- X-MS-Has-Attach: 9, 23 -- X-MS-TNEF-Correlator: 9, 23 -- Thread-Topic: Jeol 733 Microprobe 9, 23 -- Thread-Index: Acby/iWDxPLNpHquTV69e0YFSFCLwA== 9, 23 -- From: "Beavers, Roy" {rbeavers-at-mail.smu.edu} 9, 23 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com} 9, 23 -- X-OriginalArrivalTime: 18 Oct 2006 21:41:19.0350 (UTC) FILETIME=[25A01160:01C6F2FE] 9, 23 -- Content-Transfer-Encoding: 8bit 9, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9ILfJGL004997 ==============================End of - Headers==============================
==============================Original Headers============================== 22, 24 -- From kenconverse-at-qualityimages.biz Wed Oct 18 18:33:22 2006 22, 24 -- Received: from mta10.adelphia.net (mta10.adelphia.net [68.168.78.202]) 22, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9INXMNY017190 22, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 18:33:22 -0500 22, 24 -- Received: from Ken ([24.53.5.33]) by mta10.adelphia.net 22, 24 -- (InterMail vM.6.01.05.02 201-2131-123-102-20050715) with ESMTP 22, 24 -- id {20061018233318.LZLK14484.mta10.adelphia.net-at-Ken} ; 22, 24 -- Wed, 18 Oct 2006 19:33:18 -0400 22, 24 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 22, 24 -- To: {rbeavers-at-mail.smu.edu} , "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 22, 24 -- Subject: RE: [Microscopy] Jeol 733 Microprobe 22, 24 -- Date: Wed, 18 Oct 2006 19:33:14 -0400 22, 24 -- Message-ID: {002501c6f30d$c88ebd40$6401a8c0-at-Ken} 22, 24 -- MIME-Version: 1.0 22, 24 -- Content-Type: text/plain; 22, 24 -- charset="us-ascii" 22, 24 -- X-Priority: 3 (Normal) 22, 24 -- X-MSMail-Priority: Normal 22, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 22, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 22, 24 -- Importance: Normal 22, 24 -- In-Reply-To: {200610182144.k9ILiHfh009872-at-ns.microscopy.com} 22, 24 -- Content-Transfer-Encoding: 8bit 22, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9INXMNY017190 ==============================End of - Headers==============================
We are using two different couplers made by OPTEM that were designed for the specific digital camera. The couplers (containing lenses) were not inexpensive. For ports, this is the standard trinocular (camera) port on top of the oculars. Previously, it was used with 35 mm cameras and worked beautifully with film....... sigh.....
JB
} I've coupled my digital cameras to several optical microscopes in my } lab without the problem you have expressed. It sounds to me like } whatever port you are using to couple the digital camera to the } microscope has a problem. May I ask how you made the coupling? What } port are you using? What magnification are you working at? } } I actually have a coupler I use in the field when going to outside } labs that I can use to couple my digital camera to their microscopes } if the need arises. I've only had the kind of problem you are } describing when faced with a port that has some kind of problem ... } I will say at high mags, like 1000X and higher, I have run into } problems, but I have felt it is due to the pixel resolution of my } camera.... } } dj } } On Wed, 18 Oct 2006, bozzola-at-siu.edu wrote: } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
==============================Original Headers============================== 5, 21 -- From bozzola-at-siu.edu Wed Oct 18 18:39:41 2006 5, 21 -- Received: from abbmta1.siu.edu (abbmta1.siu.edu [131.230.254.205]) 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9INddWd001310 5, 21 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 18 Oct 2006 18:39:41 -0500 5, 21 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 5, 21 -- by abbmta1.siu.edu (Switch-3.1.10/Switch-3.1.10) with ESMTP id k9INdcFR026474 5, 21 -- for {Microscopy-at-msa.microscopy.com} ; Wed, 18 Oct 2006 18:39:39 -0500 (CDT) 5, 21 -- Mime-Version: 1.0 5, 21 -- X-Sender: bozzola-at-saluki-mail.siu.edu 5, 21 -- Message-Id: {p0611042ac15c6b219726-at-[131.230.177.142]} 5, 21 -- In-Reply-To: {Pine.WNT.4.64.0610181702210.3148-at-H-F1} 5, 21 -- References: {200610182047.k9IKloI7015067-at-ns.microscopy.com} 5, 21 -- {Pine.WNT.4.64.0610181702210.3148-at-H-F1} 5, 21 -- Date: Wed, 18 Oct 2006 18:39:49 -0500 5, 21 -- To: Microscopy-at-msa.microscopy.com 5, 21 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 5, 21 -- Subject: Re: [Microscopy] LM: digital camera lacking flat field 5, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 21 -- X-Spam-Score: 0.00% 5, 21 -- X-MASF: 0.00% 5, 21 -- X-Whitelist: 0.00% ==============================End of - Headers==============================
Just an update on our faulty Hitachi H-600 CRT. Our electrician, Tony Morgan, has fixed the problem. We didn't have 15V entering the CRT unit. He traced the fault back to circuit board PC-1 in the power box. The volatage regultor for fuse F6 had a "dry joint". He removed PC-1 and resoldered the pins on the voltage regulator and the fuse clips. He also replaced the fuse.
This would explain why the CRT only lasted 10 minutes each morning - as the power box heated, the slightest amount of expansion in the "dry joint" was causing the loss of continuity in the circuit.
Thanks to everyone who provided advice on this issue.
John Brealey Queen Elizabeth Hospital EM Unit Adelaide South Australia
==============================Original Headers============================== 6, 33 -- From john.brealey-at-imvs.sa.gov.au Wed Oct 18 19:01:27 2006 6, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 6, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9J01QmC016733 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 19:01:26 -0500 6, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 6, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id k9J01OJZ001259 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 09:31:24 +0930 (CST)' 6, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 6, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id k9J01O3Y001256 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 09:31:24 +0930 (CST)' 6, 33 -- Received: from localhost (scalix1.imvs.sa.gov.au [10.20.98.38]) 6, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id BC98134D3E 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 09:31:24 +0930 (CST) 6, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 6, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 6, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.38]) (amavisd-new, port 10024) 6, 33 -- with LMTP id YjK3ws2u5hLc for {Microscopy-at-MSA.Microscopy.Com} ; 6, 33 -- Thu, 19 Oct 2006 09:31:16 +0930 (CST) 6, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 6, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 8212634D63 6, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 09:30:32 +0930 (CST) 6, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 6, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 6, 33 -- Subject: Hitachi H-600 CRT Fixed 6, 33 -- Date: Thu, 19 Oct 2006 09:30:30 +0930 6, 33 -- Message-ID: {000001c6f311$9822f820$c88a140a-at-iqe36042} 6, 33 -- MIME-Version: 1.0 6, 33 -- Content-Type: text/plain; 6, 33 -- charset="us-ascii" 6, 33 -- Content-Transfer-Encoding: 7bit 6, 33 -- X-Mailer: Microsoft Office Outlook 11 6, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 6, 33 -- Thread-Index: AcbzEXNlC41UFSJlSR22Qp7NrxzA1g== ==============================End of - Headers==============================
Isabell and Dravid have considered the problem in a paper on "Resolution and sensitivity of electron backscattered diffraction in a cold field emission gun SEM" in Ultramicroscopy 67 (1997) 59-68.
Concerning the stray fields that you mention as a possible explanation they write: "Some high-resolution SEMs have a strong objective lens field which "spills" onto the specimen. This results in bending of Kikuchi lines due to the excessive magnetic field of the objective lens. This problem, however, can be easily solved by placing a small aperture of permalloy above the specimen, thus trapping the magnetic flux from the lens."
More generally, they conclude that you get a lousy signal-to-noise ratio at low voltages because low energy backscattered electrons are much less efficient in exciting the phosphor than high energy BSEs. Hence the pattern quality is degraded at low voltages.
-----Original Message----- X-from: larry-at-cymru.freewire.co.uk [mailto:larry-at-cymru.freewire.co.uk] Sent: Wednesday, October 18, 2006 10:51 PM To: j.bilde-at-risoe.dk
The sharpness of EBSD patterns decreases as the SEM kV is reduced.
Does anybody know why?
The only explanation I can come up with is stray fields. The lower kV electrons are going to be more affected by stray fields.
Regards, -- Larry Stoter
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==============================Original Headers============================== 5, 14 -- From larry-at-cymru.freewire.co.uk Wed Oct 18 15:46:12 2006 5, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 5, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9IKkBWi009630 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 15:46:12 -0500 5, 14 -- Received: from [217.154.251.81] (th6dc-217-154-251-81.dial.mistral.co.uk [217.154.251.81] (may be forged)) 5, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9IKk9DI027032 5, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 18 Oct 2006 21:46:10 +0100 5, 14 -- Mime-Version: 1.0 5, 14 -- Message-Id: {p06210200c15c4186a7b4-at-[217.154.250.196]} 5, 14 -- Date: Wed, 18 Oct 2006 21:43:02 +0100 5, 14 -- To: Microscopy-at-MSA.Microscopy.Com 5, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 5, 14 -- Subject: EBSD - kV dependence of pattern sharpness 5, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 20, 25 -- From j.bilde-at-risoe.dk Thu Oct 19 04:27:50 2006 20, 25 -- Received: from mail.risoe.dk (mail.risoe.dk [130.226.48.21]) 20, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9J9RnNC004833 20, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 19 Oct 2006 04:27:50 -0500 20, 25 -- Received: from EXCHG-VS1.risoe.dk (exchg-01.risoe.dk [10.0.0.26]) 20, 25 -- by risoe.dk (PMDF V6.2-X26 #31238) with ESMTP id {IKYU08R1H6CS84029C-at-risoe.dk} 20, 25 -- for Microscopy-at-microscopy.com; Thu, 20, 25 -- 19 Oct 2006 11:27:34 +0200 (Romance Daylight Time) 20, 25 -- Date: Thu, 19 Oct 2006 11:27:33 +0200 20, 25 -- From: j.bilde-at-risoe.dk 20, 25 -- Subject: RE: [Microscopy] EBSD - kV dependence of pattern sharpness 20, 25 -- In-reply-to: {200610182051.k9IKpMLa025119-at-ns.microscopy.com} 20, 25 -- To: larry-at-cymru.freewire.co.uk 20, 25 -- Cc: Microscopy-at-microscopy.com 20, 25 -- Message-id: {6463F256A85DC14CAB07F087A2479972AFEA69-at-EXCHG-VS1.risoe.dk} 20, 25 -- MIME-version: 1.0 20, 25 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5 20, 25 -- Content-type: text/plain; charset=iso-8859-1 20, 25 -- Thread-Topic: [Microscopy] EBSD - kV dependence of pattern sharpness 20, 25 -- Thread-Index: Acby9z96W3zLxO4HTHKxG6OpMnJ+1wAZ6fJg 20, 25 -- Content-class: urn:content-classes:message 20, 25 -- X-MS-Has-Attach: 20, 25 -- X-MS-TNEF-Correlator: 20, 25 -- Content-Transfer-Encoding: 8bit 20, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9J9RnNC004833 ==============================End of - Headers==============================
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==============================Original Headers============================== 10, 29 -- From przybylowicz-at-tlabs.ac.za Thu Oct 19 06:11:14 2006 10, 29 -- Received: from silence.tlabs.ac.za (silence.tlabs.ac.za [196.21.124.19]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9JBBB95016949 10, 29 -- for {microscopy-at-microscopy.com} ; Thu, 19 Oct 2006 06:11:12 -0500 10, 29 -- Received: from segonyane.tlabs.ac.za ([196.21.125.83]:3132 helo=przybylo) 10, 29 -- by silence.tlabs.ac.za with smtp (Exim 4.43) 10, 29 -- id 1GaVnZ-0002dw-JE 10, 29 -- for microscopy-at-microscopy.com; Thu, 19 Oct 2006 13:10:53 +0200 10, 29 -- Message-ID: {036d01c6f36f$0bb57a80$537d15c4-at-przybylo} 10, 29 -- From: "Przybylowicz Wojciech" {przybylowicz-at-tlabs.ac.za} 10, 29 -- To: {microscopy-at-microscopy.com} 10, 29 -- Subject: Post-doctoral fellow: Nuclear microprobe of thin frozen-hydrated biological specimens 10, 29 -- Date: Thu, 19 Oct 2006 13:09:23 +0200 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-Type: text/plain; 10, 29 -- charset="iso-8859-1" 10, 29 -- Content-Transfer-Encoding: 7bit 10, 29 -- X-Priority: 3 10, 29 -- X-MSMail-Priority: Normal 10, 29 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1106 10, 29 -- Disposition-Notification-To: "Przybylowicz Wojciech" {przybylowicz-at-tlabs.ac.za} 10, 29 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1106 10, 29 -- X-tlabs-Information: Please contact iThemba LABS, South Africa 10, 29 -- X-tlabs-MailScanner: Found to be clean 10, 29 -- X-tlabs-SpamCheck: not spam (whitelisted), SpamAssassin (not cached, 10, 29 -- score=-4.568, required 6, autolearn=not spam, ALL_TRUSTED -3.30, 10, 29 -- BAYES_00 -2.60, CELL_PHONE_IMPROVE 1.03, SARE_WEOFFER 0.30) 10, 29 -- X-tlabs-MailScanner-From: przybylowicz-at-tlabs.ac.za 10, 29 -- X-tlabs-MailScanner-To: microscopy-at-microscopy.com ==============================End of - Headers==============================
Hi, I'm not sure what JEOL uses on the JXA 733, but on the JXA 8600 they use an oil in that cooling circuit. Is this in part due to the fact that on the 8600 the voltage lines to the lens run inside the cooling lines? Was the 733 designed the same way? If it is the same, JEOL has the oil as:
Catalog Number Description Unit Price 423003 Lens cooling oil (per gallon) $13.96
Mike
} Date: Wed, 18 Oct 2006 18:40:35 -0500 } To: mmcheath-at-mailbox.syr.edu } From: kenconverse-at-qualityimages.biz } Reply-to: kenconverse-at-qualityimages.biz } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] RE: Jeol 733 Microprobe } X-lewp: MicroscopyListSpam NAGS } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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==============================Original Headers============================== 11, 12 -- From mmcheath-at-mailbox.syr.edu Thu Oct 19 06:16:42 2006 11, 12 -- Received: from mailer.syr.edu (mailer.syr.edu [128.230.18.29]) 11, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9JBGgVY026947 11, 12 -- for {microscopy-at-microscopy.com} ; Thu, 19 Oct 2006 06:16:42 -0500 11, 12 -- Received: from [128.230.24.90] (www.geochemistry.syr.edu) by mailer.syr.edu (LSMTP for Windows NT v1.1b) with SMTP id {0.15978AAD-at-mailer.syr.edu} ; Thu, 19 Oct 2006 7:16:41 -0400 11, 12 -- Mime-Version: 1.0 11, 12 -- Message-Id: {a06230907c15d0e5934de-at-[128.230.24.90]} 11, 12 -- Date: Thu, 19 Oct 2006 07:16:38 -0400 11, 12 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 11, 12 -- From: Michael Cheatham {mmcheath-at-mailbox.syr.edu} 11, 12 -- Subject: Fwd: [Microscopy] RE: Jeol 733 Microprobe 11, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Thanks for all the comments. The question was a general one, not related to a specific experiment/instrument, although it does seem to be a widespread observation.
A few more thoughts:
1. The reduction of interation volume at lower kVs leading to a larger proportion of the signal coming from a less than perfect surface (or contaminating surface) seems the most likely explanation. Would electrochemical polishing or glancing-incidence argon ion milling provide a better finish than colloidal silica?
2. I don't think it is related to energy spread in the electron beam - there doesn't seem to be a difference between pattern sharpness on cold FEGs compared with schottkey FEGs or W instruments.
3. While there might be distortion problems with SEMs which don't have fully contained fields, the loss of sharpness at lower kVs still occurs even on SEMs with fully contained fields and at longer working distances.
4. While I agree that the S/N ratio does deteriorate at the lowest kVs, the loss of sharpness is still apparent at kVs where this shouldn't be an issue. For example, the pattern sharpness does deteriorate in going from 30 kV to 10 kV.
5. A customer in the UK has a system where field cancellation is required for image resolution. He's going to check the patterns with and without field cancellation to see the effect - this should clarify whether stray fields have a significant effect on EBSP sharpness - I'll let the list know of the result.
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==============================Original Headers============================== 10, 14 -- From larry-at-cymru.freewire.co.uk Thu Oct 19 14:24:39 2006 10, 14 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 10, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9JJOcUu028558 10, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 14:24:38 -0500 10, 14 -- Received: from [217.154.248.147] (th1dc-217-154-248-147.dial.mistral.co.uk [217.154.248.147]) 10, 14 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9JJO5s4003860 10, 14 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 19 Oct 2006 20:24:18 +0100 10, 14 -- Mime-Version: 1.0 10, 14 -- Message-Id: {p06210200c15d79b53467-at-[217.154.251.81]} 10, 14 -- Date: Thu, 19 Oct 2006 20:18:42 +0100 10, 14 -- To: Microscopy-at-MSA.Microscopy.Com 10, 14 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 10, 14 -- Subject: Re: [Microscopy] EBSD - kV dependence of pattern sharpness 10, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Reminder! The USCB Advanced Microscopy and Digital Imaging Workshop starts on 13th of November.
Twice a year University of California at Santa Barbara runs a four and a half day workshop on modern techniques in microscopy and electronic imaging for biological research. The event is held at UC Santa Barbara's Integrated Microscopy Facility. For the second time this year the course is co-organized by Purdue University Cytometry Laboratories.
Participants will be using state-of-the art equipment from Olympus, including confocal microscopes, and modern wide-field fluorescence microscopes. Other materials include electronic cameras provided by Q-Imaging, image processing and 3D reconstruction software from Media Cybernetics, an innovative dark-field microscopy system from CytoViva, reagents from Jackson Immunoresearch, and micromanipulators/microinjectors from Eppendorf.
The Advanced Microscopy and Digital Imaging Workshop will take place at UCSB campus, in Biological Sciences II building, from 13th to 17th of November, covering everything from the basics (parts of a microscope), and specimen preparation in fluorescence microscopy to operation of confocal and two-photon systems.
You still have a chance to register at USCB course web-site.
Contact Brian Matsumoto at (805) 893-8702 for additional information.
Regards,
Bartek Rajwa
-- Bartlomiej Rajwa, PhD Purdue University Cytometry Laboratories Bindley Bioscience Center (BIND) 1203 W. State Street Purdue University, West Lafayette, IN 47907-2057 tel. (765) 494 0757
==============================Original Headers============================== 10, 20 -- From rajwa-at-flowcyt.cyto.purdue.edu Thu Oct 19 16:42:58 2006 10, 20 -- Received: from flowcyt.cyto.purdue.edu (flowcyt.cyto.purdue.edu [128.210.61.51]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9JLgwXE011135 10, 20 -- for {microscopy-at-MSA.Microscopy.com} ; Thu, 19 Oct 2006 16:42:58 -0500 10, 20 -- Received: from [128.210.61.74] (spengler.cyto.purdue.edu [128.210.61.74]) 10, 20 -- (authenticated bits=0) 10, 20 -- by flowcyt.cyto.purdue.edu (8.12.10/8.12.10) with ESMTP id k9JLgjXM038292; 10, 20 -- Thu, 19 Oct 2006 17:42:45 -0400 (EDT) 10, 20 -- (envelope-from rajwa-at-flowcyt.cyto.purdue.edu) 10, 20 -- Message-ID: {4537F155.3090200-at-flowcyt.cyto.purdue.edu} 10, 20 -- Date: Thu, 19 Oct 2006 17:42:45 -0400 10, 20 -- From: Bartek Rajwa {rajwa-at-flowcyt.cyto.purdue.edu} 10, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.8.0.7) Gecko/20060909 Thunderbird/1.5.0.7 Mnenhy/0.7.4.0 10, 20 -- MIME-Version: 1.0 10, 20 -- To: CONFOCAL-at-LISTSERV.BUFFALO.EDU, microscopy-at-MSA.Microscopy.com 10, 20 -- Subject: Microscopy course at Santa Barbara - last announcement 10, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 20 -- Content-Transfer-Encoding: 7bit 10, 20 -- X-Spam-Score: -4.9 () BAYES_00 10, 20 -- X-Scanned-By: MIMEDefang 2.39 ==============================End of - Headers==============================
You need to define "sharpness." What I think you are seeing is reduction of S/N ratio. The Kikuchi patterns are generated by a point source. Thus, moving the phosphor screen of the camera towards or away from the specimen will change the width of the patterns. Likewise, changing WD will also have a similar effect. Higher KV will provide deeper penetration of the specimen (~50nm) and will produce good S/N. So it seems that what you are talking about is contrast or definition of lines. It is not "sharpness."
Ideally, the PC should be about 1/3 down from the top of the scintillator disc. If you are off, that will degrade the pattern quality. Also be sure to set your analysis WD for the WD actually used.
gary g.
At 01:47 PM 10/18/2006, you wrote:
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==============================Original Headers============================== 9, 20 -- From gary-at-gaugler.com Thu Oct 19 19:44:01 2006 9, 20 -- Received: from qsmtp4.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 k9K0i03P024610 9, 20 -- for {microscopy-at-microscopy.com} ; Thu, 19 Oct 2006 19:44:01 -0500 9, 20 -- Received: (qmail 11825 invoked from network); 19 Oct 2006 17:40:46 -0700 9, 20 -- Received: by simscan 1.1.0 ppid: 11822, pid: 11823, t: 0.1686s 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 qsmtp4 with SMTP; 19 Oct 2006 17:40:45 -0700 9, 20 -- Message-Id: {7.0.1.0.2.20061019173857.024fc870-at-gaugler.com} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 20 -- Date: Thu, 19 Oct 2006 17:44:43 -0700 9, 20 -- To: larry-at-cymru.freewire.co.uk 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] EBSD - kV dependence of pattern sharpness 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200610182047.k9IKlUr3014034-at-ns.microscopy.com} 9, 20 -- References: {200610182047.k9IKlUr3014034-at-ns.microscopy.com} 9, 20 -- Mime-Version: 1.0 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Helen, No-one seems to have picked up your query, so I'll have a go (even though I'm not a STEM expert..) First, there's not really enough information in your post to give an answer, it would help a lot if you could find a way to let others see the images (as was done recently for a question on backscattered imaging). Second, I believe that the contrast in HAADF-STEM is dominated by differences in atomic number, although strain contrast can be seen as well. There are differences depending upon detector geometry, I have heard that many HAADF detectors are not really 'high angle' (where you detect incoherent scattering and the image is dominated by atomic number); collecting electrons scattered at lower angles gives a stronger signal but at the expense of some coherent scattering effects appearing in your image too (e.g. strain contrast, stacking fault fringes etc.). So your idea of looking for a compositional difference at the boundary is a good one. However, the signal-to-noise ratio for EDX is several orders of magnitude worse for EDX than for HAADF, so there are lots of things you can see in HAADF which you can't distinguish in EDX maps. EELS has a much better S-N ratio and is worth a try. Alternatively you can look at the same defect in HREM. Third, why should a defect which is visible edge-on away from a zone axis (you don't say which crystallographic plane) become less visible when viewed along a zone axis (presumably still seen edge-on?)? The short answer is I don't know, I guess you are picking up coherent scattering effects. It does seem that people generally ignore dynamical electron diffraction in HAADF images (it makes interpretation so much easier), but I have a feeling that you can't get away from those Bloch waves so easily..
Hope this helps
Richard.
P.S. For anyone who isn't a materials TEM person and wants to know, the acronyms are: TEM: transmission electron microscopy STEM: scanning transmission electron microscopy HAADF: High angle annular dark field EDX: Energy dispersive X-ray spectroscopy EELS: Electron energy loss spectroscopy TLA: three letter acronym
-----Original Message----- X-from: hx213-at-cam.ac.uk [mailto:hx213-at-cam.ac.uk] Sent: 18 October 2006 15:05 To: Richard Beanland
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Email: hx213-at-cam.ac.uk Name: Huixin Xiu
Organization: University of Cambridge
Title-Subject: [Filtered] About the defect contrast in HAADF image
Question: Dear Microscopists,
I observed a strong defect ( possibly inversion domains) contrast in STEM-HAADF image at edge-on condition in GaN. The contrast also exists along {11-20} zone axis, but it is weaker than edge-on condition. I tried to use EDX to see whether any chemical change across the defect, but didn't find obvious change. Could anybody have similar observations or have any idea of the contrast from? Thanks a lot.
==============================Original Headers============================== 8, 12 -- From zaluzec-at-microscopy.com Wed Oct 18 09:03:08 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 k9IE36eN030232 8, 12 -- for {microscopy-at-microscopy.com} ; Wed, 18 Oct 2006 09:03:07 -0500 8, 12 -- Mime-Version: 1.0 8, 12 -- X-Sender: (Unverified) 8, 12 -- Message-Id: {p06110404c15be4886d2d-at-[206.69.208.22]} 8, 12 -- Date: Wed, 18 Oct 2006 09:03:05 -0500 8, 12 -- To: microscopy-at-microscopy.com 8, 12 -- From: hx213-at-cam.ac.uk (by way of MicroscopyListserver) 8, 12 -- Subject: viaWWW: defect contrast in HAADF image 8, 12 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
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==============================Original Headers============================== 21, 32 -- From richard.beanland-at-bookham.com Fri Oct 20 03:44:55 2006 21, 32 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 21, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9K8is1p012642 21, 32 -- for {microscopy-at-microscopy.com} ; Fri, 20 Oct 2006 03:44:54 -0500 21, 32 -- X-VirusChecked: Checked 21, 32 -- X-Env-Sender: richard.beanland-at-bookham.com 21, 32 -- X-Msg-Ref: server-3.tower-78.messagelabs.com!1161333893!43195126!1 21, 32 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 21, 32 -- X-Originating-IP: [213.249.209.179] 21, 32 -- Received: (qmail 20201 invoked from network); 20 Oct 2006 08:44:53 -0000 21, 32 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 21, 32 -- by server-3.tower-78.messagelabs.com with SMTP; 20 Oct 2006 08:44:53 -0000 21, 32 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 21, 32 -- Fri, 20 Oct 2006 09:48:29 +0100 21, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 21, 32 -- Content-class: urn:content-classes:message 21, 32 -- MIME-Version: 1.0 21, 32 -- Content-Type: text/plain; 21, 32 -- charset="us-ascii" 21, 32 -- Subject: RE: [Microscopy] viaWWW: defect contrast in HAADF image 21, 32 -- Date: Fri, 20 Oct 2006 09:47:54 +0100 21, 32 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E243484-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 21, 32 -- X-MS-Has-Attach: 21, 32 -- X-MS-TNEF-Correlator: 21, 32 -- Thread-Topic: [Microscopy] viaWWW: defect contrast in HAADF image 21, 32 -- Thread-Index: Acbyv2hfQAJHJ0U8RtuU1/0JXVon/QBYOYrg 21, 32 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 21, 32 -- To: {hx213-at-cam.ac.uk} 21, 32 -- Cc: {microscopy-at-microscopy.com} 21, 32 -- X-OriginalArrivalTime: 20 Oct 2006 08:48:29.0259 (UTC) FILETIME=[83B931B0:01C6F424] 21, 32 -- Content-Transfer-Encoding: 8bit 21, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9K8is1p012642 ==============================End of - Headers==============================
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If anyone uses this software and wishes to aid in some trouble shooting, i'd greatly appreciate it.
I'm not as familiar with the software as i would like to be, and i'm pretty sure my problems are software based. Atleast i'm hoping and praying its not the detector!
I can get in to more detail upon request, but for now... Our EDS hasn't been used in over a month and i've never been the primary operator. Upon getting it back up and running, i try to aquire a spectra. It goes straight to 100% deadtime and 0 counts then stops attempting to continue. I had it set for a 180 second run, but yet the finish time stays at 180, and where it says the elapsed time, there is some really big number.
I don't think its the detector, because it's always been well kept and full of LN.
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Email: martin.roe-at-nottingham.ac.uk Name: Martin Roe
Organization: Nottingham University
Title-Subject: [Filtered] Kevex DeltaPlus EDX system -FREE to collect
Question: I was about to put a Kevex DeltaPlus (Fisons Instruments) EDX system in the skip but then thought it may be useful to someone. The serial no of the system is 5500067-0135 (made in USA). There is a computer with 3.5 and 5 inch floppy drives (model PE202F-2; serial no 38151); a keyboard; Okidata Microline-320 9 pin printer and a Fisons Instruments schematics + wiring diagrams manual.Please note there is NO EDX detector. The above items are free to collect but you must arrange shipping with an appropriate courier. First come first served. I am in Nottingham in the UK. By the way, it was never actually used by anyone at this University as it came from elsewhere along with other equipment, but as far as I know it was in a working order when last used. Could be useful for spares for anyone with the same system. Martin Roe
1. turn off the chamberscope if on. fix the problem? if not 2. check the time constant (Acquire--} Setup Pulse processing --} if high (100, 50), set to low (e.g. 3) microsec. fix the problem? if not 3. turn down the beam current ("spot size").
call me that none of that works. 608 265-4798
john
-- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
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==============================Original Headers============================== 6, 25 -- From johnf-at-geology.wisc.edu Fri Oct 20 09:14:18 2006 6, 25 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 6, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9KEEIsZ018195 6, 25 -- for {microscopy-at-microscopy.com} ; Fri, 20 Oct 2006 09:14:18 -0500 6, 25 -- Received: from localhost (localhost [127.0.0.1]) 6, 25 -- by localhost (Postfix) with ESMTP id DD95420D02; 6, 25 -- Fri, 20 Oct 2006 09:14:13 -0500 (CDT) 6, 25 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 6, 25 -- by localhost (geology.wisc.edu [127.0.0.1]) (amavisd-new, port 10024) 6, 25 -- with ESMTP id 11929-03; Fri, 20 Oct 2006 09:14:10 -0500 (CDT) 6, 25 -- Received: from [144.92.206.57] (beamer.geology.wisc.edu [144.92.206.57]) 6, 25 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 6, 25 -- (No client certificate requested) 6, 25 -- by ice.geology.wisc.edu (Postfix) with ESMTP id 8E54220D26; 6, 25 -- Fri, 20 Oct 2006 09:14:10 -0500 (CDT) 6, 25 -- Mime-Version: 1.0 6, 25 -- Message-Id: {p06230905c15e897a452f-at-[144.92.206.57]} 6, 25 -- In-Reply-To: {200610201329.k9KDTg4Y012533-at-ns.microscopy.com} 6, 25 -- References: {200610201329.k9KDTg4Y012533-at-ns.microscopy.com} 6, 25 -- Date: Fri, 20 Oct 2006 09:14:28 -0500 6, 25 -- To: burrmich-at-msu.edu, microscopy-at-microscopy.com 6, 25 -- From: John Fournelle {johnf-at-geology.wisc.edu} 6, 25 -- Subject: Re: [Microscopy] viaWWW: EDS software question (WINEDS) 6, 25 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 25 -- X-Virus-Scanned: amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
HI, I wonder any body here has experience on using the two materials Hexamethyldisilizane (HMDS) and tetramethylsilane (TMS) as alternatives to the conventional CO2 CPD? Jun He SEM lab UNMC, Omaha, NE
==============================Original Headers============================== 1, 21 -- From junhe-at-unmc.edu Fri Oct 20 09:38:04 2006 1, 21 -- Received: from zixvpm01.unmc.edu (zixvpm01.unmc.edu [192.198.54.126]) 1, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9KEc4Xw029044 1, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Oct 2006 09:38:04 -0500 1, 21 -- Received: from zixvpm01.unmc.edu (ZixVPM [127.0.0.1]) 1, 21 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 196DD4C0D9 1, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Oct 2006 09:38:04 -0500 (CDT) 1, 21 -- Received: from unmcnotes05.unmc.edu (unknown [10.9.2.163]) 1, 21 -- by zixvpm01.unmc.edu (Proprietary) with ESMTP id 523BA4C116 1, 21 -- for {Microscopy-at-Microscopy.Com} ; Fri, 20 Oct 2006 09:38:02 -0500 (CDT) 1, 21 -- To: Microscopy-at-Microscopy.Com 1, 21 -- Subject: Using HMDS and TMS to replace CO2 CPD 1, 21 -- MIME-Version: 1.0 1, 21 -- X-Mailer: Lotus Notes Release 7.0.1 January 17, 2006 1, 21 -- Message-ID: {OF0B3D7122.0AB7A864-ON8625720D.00504BD0-8625720D.005060A1-at-unmc.edu} 1, 21 -- From: Jun He {junhe-at-unmc.edu} 1, 21 -- Date: Fri, 20 Oct 2006 09:37:56 -0500 1, 21 -- X-MIMETrack: Serialize by Router on UNMCNOTES05.UNMC.EDU/Servers/UNEBR at 10/20/2006 09:38:02 1, 21 -- AM, 1, 21 -- Serialize complete at 10/20/2006 09:38:02 AM 1, 21 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
I'm using WINEDS software version 4.0 every day more than 8 years without any problems. We using this system with two EDX detector Edax Si(Li) Be window and now with Gresham Sirius SUTW window. One question, was detector over this time filled with the LN2.
Henrik
burrmich-at-msu.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://microscopy.com/MicroscopyListserver/MLFormMail.html } --------------------------------------------------------------------------- } Remember this posting is most likely not from a Subscriber, so when replying } please copy both burrmich-at-msu.edu as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: burrmich-at-msu.edu } Name: Mike } } Title-Subject: [Filtered] EDS software question (WINEDS) } } Question: Hello all, } } If anyone uses this software and wishes to aid in some trouble shooting, i'd greatly appreciate it. } } I'm not as familiar with the software as i would like to be, and i'm pretty sure my problems are software based. Atleast i'm hoping and praying its not the detector! } } I can get in to more detail upon request, but for now... Our EDS hasn't been used in over a month and i've never been the primary operator. Upon getting it back up and running, i try to aquire a spectra. It goes straight to 100% deadtime and 0 counts then stops attempting to continue. I had it set for a 180 second run, but yet the finish time stays at 180, and where it says the elapsed time, there is some really big number. } } I don't think its the detector, because it's always been well kept and full of LN. } } Any help would be greatly appreciated. } } Thanks, } Mike } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 11, 12 -- From zaluzec-at-microscopy.com Fri Oct 20 08:21:47 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 k9KDLkDp028486 } 11, 12 -- for {microscopy-at-microscopy.com} ; Fri, 20 Oct 2006 08:21:46 -0500 } 11, 12 -- Mime-Version: 1.0 } 11, 12 -- X-Sender: (Unverified) } 11, 12 -- Message-Id: {p06110400c15e7dde27d1-at-[206.69.208.22]} } 11, 12 -- Date: Fri, 20 Oct 2006 08:21:45 -0500 } 11, 12 -- To: microscopy-at-microscopy.com } 11, 12 -- From: burrmich-at-msu.edu (by way of MicroscopyListserver) } 11, 12 -- Subject: viaWWW: EDS software question (WINEDS) } 11, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== } } } __________ NOD32 1.1819 (20061020) Information __________ } } This message was checked by NOD32 antivirus system. } http://www.eset.com } } } }
-- Henrik Kaker Ph.D. Metal Ravne d.o.o. SEM-EDS Laboratory Koroska cesta 14 SI-2390 Ravne Slovenia Phone: +386 2 870 7076 GSM: +386 31 380 875 http://www2.arnes.si/~sgszmera1/index.html
==============================Original Headers============================== 7, 27 -- From Henrik.Kaker-at-guest.arnes.si Fri Oct 20 11:40:17 2006 7, 27 -- Received: from avs1.arnes.si (avs1.arnes.si [193.2.1.74]) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9KGeGnR009961 7, 27 -- for {microscopy-at-microscopy.com} ; Fri, 20 Oct 2006 11:40:17 -0500 7, 27 -- Received: from localhost (avs1.arnes.si [193.2.1.74]) 7, 27 -- by avs1.arnes.si (Postfix) with ESMTP id 268A636A676; 7, 27 -- Fri, 20 Oct 2006 18:40:16 +0200 (CEST) 7, 27 -- Received: from avs1.arnes.si ([193.2.1.74]) 7, 27 -- by localhost (avs1.arnes.si [193.2.1.74]) (amavisd-new, port 10024) 7, 27 -- with ESMTP id 05585-17; Fri, 20 Oct 2006 18:40:15 +0200 (CEST) 7, 27 -- Received: from [89.212.22.21] (89-212-22-21.static.dsl.t-2.net [89.212.22.21]) 7, 27 -- by avs1.arnes.si (Postfix) with ESMTP id D9F8936A671; 7, 27 -- Fri, 20 Oct 2006 18:40:15 +0200 (CEST) 7, 27 -- Message-ID: {4538FBFA.8020503-at-guest.arnes.si} 7, 27 -- Date: Fri, 20 Oct 2006 18:40:26 +0200 7, 27 -- From: Henrik Kaker {Henrik.Kaker-at-guest.arnes.si} 7, 27 -- Reply-To: Henrik.Kaker-at-guest.arnes.si 7, 27 -- Organization: SEM-EDS Lab 7, 27 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 7, 27 -- MIME-Version: 1.0 7, 27 -- To: burrmich-at-msu.edu, "Microscopy Listserver" {microscopy-at-microscopy.com} 7, 27 -- Subject: Re: [Microscopy] viaWWW: EDS software question (WINEDS) 7, 27 -- References: {200610201322.k9KDMA22028909-at-ns.microscopy.com} 7, 27 -- In-Reply-To: {200610201322.k9KDMA22028909-at-ns.microscopy.com} 7, 27 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 27 -- Content-Transfer-Encoding: 7bit 7, 27 -- X-Virus-Scanned: by amavisd-new at arnes.si ==============================End of - Headers==============================
A postdoctoral position in advanced materials characterization using TEM/SEM/XPS is available at the University of Louisville, Institute for Advanced Materials & Renewable Energy (IAM-RE) Louisville KY 40292. Applicants should have a Ph.D. degree in Engineering or sciences in a related field with extensive and demonstrated hands-on experience using either transmission electron microscopy or surface science tools.
Facilities at IAM-RE include a TECNAI F20 XTWIN STEM/TEM with GIF 2002, a Nova 600 FEG SEM, and a VG Scientific MultiLab for XPS, Auger, STM, Raman/PL, FTIR, XRD etc.
The position is available immediately for one year and can be extended to second year. Interested candidates should send a curriculum vitae, publication list, and the names of three references with their contact addresses to: mark.shreck-at-louisville.edu
University of Louisville is an Equal Opportunity/Affirmative Action Employer and is strongly committed to building diversity within its community
Thank you for your attention
==============================Original Headers============================== 6, 26 -- From z0chen09-at-louisville.edu Fri Oct 20 12:05:13 2006 6, 26 -- Received: from erouter2.it-datacntr.louisville.edu (erouter2.it-datacntr.louisville.edu [136.165.237.35]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9KH5Cru021026 6, 26 -- for {microscopy-at-microscopy.com} ; Fri, 20 Oct 2006 12:05:13 -0500 6, 26 -- Received: from PostX2 (postx2.it-servers.louisville.edu [136.165.233.129]) 6, 26 -- by erouter2.it-datacntr.louisville.edu (Postfix) with ESMTP id 03E496C26A 6, 26 -- for {microscopy-at-microscopy.com} ; Fri, 20 Oct 2006 13:05:12 -0400 (EDT) 6, 26 -- Received: from savgw-out.louisville.edu ([136.165.234.42]) 6, 26 -- by PostX2 (PostX Enterprise 6.0.1 SMTP Adapter) with SMTP ID 787 6, 26 -- for {microscopy-at-microscopy.com} ; 6, 26 -- Fri, 20 Oct 2006 12:52:13 -0400 (EDT) 6, 26 -- Received: from gwise.louisville.edu ([136.165.232.131]) 6, 26 -- by savgw-out.louisville.edu (SAVSMTP 3.1.7.47) with SMTP id M2006102013030426847 6, 26 -- for {microscopy-at-microscopy.com} ; Fri, 20 Oct 2006 13:03:04 -0400 6, 26 -- Received: from sgate-MTA by gwise.louisville.edu 6, 26 -- with Novell_GroupWise; Fri, 20 Oct 2006 13:05:10 -0400 6, 26 -- Message-Id: {4538C980.B28C.005C.0-at-gwise.louisville.edu} 6, 26 -- X-Mailer: Novell GroupWise Internet Agent 7.0.1 6, 26 -- Date: Fri, 20 Oct 2006 13:05:01 -0400 6, 26 -- From: "Zhiqiang Chen" {z0chen09-at-louisville.edu} 6, 26 -- To: {microscopy-at-microscopy.com} 6, 26 -- Subject: Postdoctoral Position Available 6, 26 -- Mime-Version: 1.0 6, 26 -- Content-Type: text/plain; charset=US-ASCII 6, 26 -- Content-Transfer-Encoding: 7bit 6, 26 -- Content-Disposition: inline ==============================End of - Headers==============================
} You need to define "sharpness." What I think you are } seeing is reduction of S/N ratio. The Kikuchi patterns } are generated by a point source. Thus, moving the phosphor } screen of the camera towards or away from the specimen will } change the width of the patterns. Likewise, changing WD } will also have a similar effect. Higher KV will provide } deeper penetration of the specimen (~50nm) and will produce } good S/N. So it seems that what you are talking about is } contrast or definition of lines. It is not "sharpness." } } Ideally, the PC should be about 1/3 down from the top of } the scintillator disc. If you are off, that will degrade } the pattern quality. Also be sure to set your analysis } WD for the WD actually used. } } gary g.
I agree that there is a need to separate 'sharpness' from 'contrast'. And 'contrast' does relate to S/N and probe current.
However, there does still seem to be a loss of sharpness at lower kV, as opposed to S/N.
I think the likely explanation relates to surface damage and the smaller interaction volume at lower kV.
Clearly, sample preparation is critical for good EBSPs - and this means a sequence of steps. Any individual process will generate its own surface 'damage'. The key is that each step should remove the damage from the preceeding step while generating a significantly thinner damage layer.
It is also necessary to understand the mechanical properties of the materials being prepared. My personal experience is with austenitic stainless steels, where mechanical processing can easily generate dense dislocation tangles (work hardening) several 100 ums deep. Polishing with colloidal silica or ion milling won't remove even half of the damage layer from the initial mechanical preparation - which is why I spent a lot of time with spark cutter systems .... -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
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==============================Original Headers============================== 7, 17 -- From larry-at-cymru.freewire.co.uk Fri Oct 20 14:48:16 2006 7, 17 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 7, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9KJmEUp002496 7, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 20 Oct 2006 14:48:15 -0500 7, 17 -- Received: from [217.154.249.175] (th2dc-217-154-249-175.dial.mistral.co.uk [217.154.249.175]) 7, 17 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9KJm5gb018704; 7, 17 -- Fri, 20 Oct 2006 20:48:10 +0100 7, 17 -- Mime-Version: 1.0 7, 17 -- Message-Id: {p06210201c15ed3de437f-at-[217.154.248.147]} 7, 17 -- In-Reply-To: {7.0.1.0.2.20061019173857.024fc870-at-gaugler.com} 7, 17 -- References: {200610182047.k9IKlUr3014034-at-ns.microscopy.com} 7, 17 -- {7.0.1.0.2.20061019173857.024fc870-at-gaugler.com} 7, 17 -- Date: Fri, 20 Oct 2006 20:43:17 +0100 7, 17 -- To: Gary Gaugler {gary-at-gaugler.com} , Microscopy-at-MSA.Microscopy.Com 7, 17 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 7, 17 -- Subject: Re: [Microscopy] EBSD - kV dependence of pattern sharpness 7, 17 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
A couple of other things came to mind after my first posting.
A real killer of patterns is oxidation. With EBSD data coming from only about 50nm deep, it does not take much to reach a point of no patterns at all. I baselined Al on Silicon wafers and after polishing (mechanical down to .02u colloidal silica) the patterns slowly diminished over three days after exposure to air. Then, in about an additional four days, no patterns.
No doubt at all that specimen prep is critical to good patterns and even getting patterns at all. And indeed, different types of specimens require different prep methods. This is learned the hard way.
Another factor is the binning value of the CCD. This works against lower KV in that the patterns can be very sharp without binning but the fps are very low. Like 3-5fps is not uncommon, but the images are great. This is mostly useful for collecting a single nice looking pattern for publication or reporting.
Also, at lower KV you will need to increase camera gain (contrast). This tends to make the final pattern un-even from top to bottom. Doing an initial good adjustment of gain and brightness level along with background subtraction helps a lot in this respect. In some cases, adjusting WD will even out the overall video signal.
Another issue is whether the camera/phosphor is tilted or horizontal. Phosphor should be tilted up, camera down. This way, the incidence angle of the pattern for each point collected is symmetrical. I'm sure that the EBSD gurus can explain it better than I can.
If you can put a link to a pix of what you are working with or one that shows your concern, that would help.
gary g.
At 12:50 PM 10/20/2006, you wrote:
} I agree that there is a need to separate 'sharpness' from 'contrast'. } And 'contrast' does relate to S/N and probe current. } } However, there does still seem to be a loss of sharpness at lower kV, } as opposed to S/N. } } I think the likely explanation relates to surface damage and the } smaller interaction volume at lower kV. } } Clearly, sample preparation is critical for good EBSPs - and this } means a sequence of steps. Any individual process will generate its } own surface 'damage'. The key is that each step should remove the } damage from the preceeding step while generating a significantly } thinner damage layer. } } It is also necessary to understand the mechanical properties of the } materials being prepared. My personal experience is with austenitic } stainless steels, where mechanical processing can easily generate } dense dislocation tangles (work hardening) several 100 ums deep. } Polishing with colloidal silica or ion milling won't remove even half } of the damage layer from the initial mechanical preparation - which } is why I spent a lot of time with spark cutter systems .... } -- } Larry Stoter } JEOL (UK) Ltd } tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoleuro.com
==============================Original Headers============================== 13, 20 -- From gary-at-gaugler.com Fri Oct 20 16:35:56 2006 13, 20 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k9KLZuDM015406 13, 20 -- for {microscopy-at-microscopy.com} ; Fri, 20 Oct 2006 16:35:56 -0500 13, 20 -- Received: (qmail 24392 invoked from network); 20 Oct 2006 14:35:56 -0700 13, 20 -- Received: by simscan 1.1.0 ppid: 24389, pid: 24390, t: 0.1685s 13, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 13, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 13, 20 -- by qsmtp2 with SMTP; 20 Oct 2006 14:35:55 -0700 13, 20 -- Message-Id: {7.0.1.0.2.20061020141415.025cd798-at-gaugler.com} 13, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 20 -- Date: Fri, 20 Oct 2006 14:35:35 -0700 13, 20 -- To: larry-at-cymru.freewire.co.uk 13, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 20 -- Subject: Re: [Microscopy] Re: EBSD - kV dependence of pattern sharpness 13, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 20 -- In-Reply-To: {200610201950.k9KJoThL005525-at-ns.microscopy.com} 13, 20 -- References: {200610201950.k9KJoThL005525-at-ns.microscopy.com} 13, 20 -- Mime-Version: 1.0 13, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Just a test to see if this gets past the spam filter. -- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 1, 20 -- From oshel1pe-at-cmich.edu Mon Oct 23 07:03:14 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 k9NC3EG1020405 1, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 23 Oct 2006 07:03:14 -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 k9NCXxNC009403 1, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 23 Oct 2006 08:34:10 -0400 1, 20 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 1, 20 -- Mon, 23 Oct 2006 08:03:12 -0400 1, 20 -- Mime-Version: 1.0 1, 20 -- Message-Id: {f06230900c1625fe0899b-at-[141.209.160.249]} 1, 20 -- Date: Mon, 23 Oct 2006 08:03:11 -0400 1, 20 -- To: Microscopy-at-microscopy.com 1, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 1, 20 -- Subject: test message, do not read 1, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 1, 20 -- X-OriginalArrivalTime: 23 Oct 2006 12:03:13.0136 (UTC) FILETIME=[37189700:01C6F69B] 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==============================
Eleven videos from MAS have now been added to the MSA collection. These are numbers 288-298. The videos are from a 1986 Presidential Symposium on the History of Microanalysis. These were converted to DVD thanks to Paul Carpenter (and NASA). The full titles can be found at the MSA web site (http://www.microscopy.org/) Under Reference and Education. If there is an MAS list server, would someone please forward this information to that list. Greg Erdos gwe-at-ufl.edu 352-466-0843 Micanopy Florida
==============================Original Headers============================== 2, 27 -- From gwe-at-ufl.edu Mon Oct 23 08:44:31 2006 2, 27 -- Received: from imf23aec.mail.bellsouth.net (imf23aec.mail.bellsouth.net [205.152.59.71]) 2, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9NDiOEM000530 2, 27 -- for {microscopy-at-microscopy.org} ; Mon, 23 Oct 2006 08:44:30 -0500 2, 27 -- Received: from ibm66aec.bellsouth.net ([68.220.152.158]) 2, 27 -- by imf23aec.mail.bellsouth.net with ESMTP 2, 27 -- id {20061023134418.HDYN12526.imf23aec.mail.bellsouth.net-at-ibm66aec.bellsouth.net} 2, 27 -- for {microscopy-at-microscopy.org} ; Mon, 23 Oct 2006 09:44:18 -0400 2, 27 -- Received: from ufcdd0b59c8a4d ([68.220.152.158]) by ibm66aec.bellsouth.net 2, 27 -- with SMTP 2, 27 -- id {20061023134418.VSGV6138.ibm66aec.bellsouth.net-at-ufcdd0b59c8a4d} 2, 27 -- for {microscopy-at-microscopy.org} ; Mon, 23 Oct 2006 09:44:18 -0400 2, 27 -- Message-ID: {004101c6f6a9$5195b730$9e98dc44-at-ufcdd0b59c8a4d} 2, 27 -- From: "greg erdos" {gwe-at-ufl.edu} 2, 27 -- To: {microscopy-at-microscopy.org} 2, 27 -- Subject: new videos 2, 27 -- Date: Mon, 23 Oct 2006 09:44:10 -0400 2, 27 -- MIME-Version: 1.0 2, 27 -- Content-Type: text/plain; 2, 27 -- format=flowed; 2, 27 -- charset="iso-8859-1"; 2, 27 -- reply-type=original 2, 27 -- Content-Transfer-Encoding: 7bit 2, 27 -- X-Priority: 3 2, 27 -- X-MSMail-Priority: Normal 2, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 2, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
Greg Erdos gwe-at-ufl.edu 352-466-0843 Micanopy Florida ----- Original Message ----- X-from: "greg erdos" {gwe-at-ufl.edu} To: {microscopy-at-microscopy.org} Sent: Monday, October 23, 2006 9:44 AM
I am posting this for a friend. Facilities coordinator at the University of Alabama, See: http://www.as.ua.edu/biology/opticalcoordinator.htm Greg Erdos gwe-at-ufl.edu 352-466-0843 Micanopy Florida
==============================Original Headers============================== 2, 27 -- From gwe-at-ufl.edu Mon Oct 23 17:05:07 2006 2, 27 -- Received: from imf24aec.mail.bellsouth.net (imf24aec.mail.bellsouth.net [205.152.59.72]) 2, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9NM57a8004212 2, 27 -- for {microscopy-at-microscopy.com} ; Mon, 23 Oct 2006 17:05:07 -0500 2, 27 -- Received: from ibm70aec.bellsouth.net ([68.220.152.158]) 2, 27 -- by imf24aec.mail.bellsouth.net with ESMTP 2, 27 -- id {20061023220507.QFMI11320.imf24aec.mail.bellsouth.net-at-ibm70aec.bellsouth.net} 2, 27 -- for {microscopy-at-microscopy.com} ; Mon, 23 Oct 2006 18:05:07 -0400 2, 27 -- Received: from ufcdd0b59c8a4d ([68.220.152.158]) by ibm70aec.bellsouth.net 2, 27 -- with SMTP 2, 27 -- id {20061023220506.DJRF28704.ibm70aec.bellsouth.net-at-ufcdd0b59c8a4d} 2, 27 -- for {microscopy-at-microscopy.com} ; Mon, 23 Oct 2006 18:05:06 -0400 2, 27 -- Message-ID: {003601c6f6ef$47812360$9e98dc44-at-ufcdd0b59c8a4d} 2, 27 -- From: "greg erdos" {gwe-at-ufl.edu} 2, 27 -- To: {microscopy-at-microscopy.com} 2, 27 -- Subject: Job Opening 2, 27 -- Date: Mon, 23 Oct 2006 18:04:57 -0400 2, 27 -- MIME-Version: 1.0 2, 27 -- Content-Type: text/plain; 2, 27 -- format=flowed; 2, 27 -- charset="iso-8859-1"; 2, 27 -- reply-type=original 2, 27 -- Content-Transfer-Encoding: 7bit 2, 27 -- X-Priority: 3 2, 27 -- X-MSMail-Priority: Normal 2, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 2, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
The Biostructure Core Facility at The Forsyth Institute, a world-renowned biomedical research institution, provides essential histological and structural analysis support for its principal investigators who are engaged in NIH-supported research related to oral biology and craniofacial development. A staff position is currently available within this facility. Candidates with appropriate training and significant biomedical research experience in confocal microscopy, histology, electron microscopy (SEM, TEM) and tissue sample preparation are encouraged to apply. The successful candidate will have the opportunity to play a critical role in the pursuit of research goals. Opportunities to present findings at national meetings and publish results. Forsyth is an academic organization and offers competitive salaries and excellent fringe benefits. Send resume to: Human Resources The Forsyth Institute 140 Fenway, Boston 02115 humanresourcesr-at-forsyth.org Affirmative Action/Equal Oppty Employer M/F/H/V
==============================Original Headers============================== 6, 20 -- From EBeniash-at-forsyth.org Tue Oct 24 08:39:47 2006 6, 20 -- Received: from mail.forsyth.org (mail.forsyth.org [65.196.9.148]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9ODdkZ9031300 6, 20 -- for {microscopy-at-microscopy.com} ; Tue, 24 Oct 2006 08:39:46 -0500 6, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 6, 20 -- Content-class: urn:content-classes:message 6, 20 -- MIME-Version: 1.0 6, 20 -- Content-Type: text/plain; 6, 20 -- charset="iso-8859-1" 6, 20 -- Subject: EM/Histology Research Position 6, 20 -- Date: Tue, 24 Oct 2006 09:39:40 -0400 6, 20 -- Message-ID: {0B76D1616988384AB8F15902BF5234F9CC7AE2-at-kamino.ohsis2.forsyth.org} 6, 20 -- X-MS-Has-Attach: 6, 20 -- X-MS-TNEF-Correlator: 6, 20 -- Thread-Topic: EM/Histology Research Position 6, 20 -- Thread-Index: Acb3cdryv/Axn1FEQQSYfOjhGlai4w== 6, 20 -- From: "Beniash, Elia" {EBeniash-at-forsyth.org} 6, 20 -- To: {microscopy-at-microscopy.com} 6, 20 -- Content-Transfer-Encoding: 8bit 6, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9ODdkZ9031300 ==============================End of - Headers==============================
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Title-Subject: [Filtered] PhD position in Materials Science
Question:
A PhD position is opened in the field of Materials Science in the Laboratory for Materials Behaviour at Paul Scherrer Institut, with the objective of examining the mechanism of oxidation of zirconium based alloys using AFM, SEM, and TEM.
The successful candidate should have a Masters / diploma degree in materials science or related fields and should have a basic knowledge of structural metallurgy. Good computational skills and a basic understanding of crystallography are required. Experience with TEM and SEM and / or AFM will be a great advantage.
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] Emitech K775 and K250 Manuals
Question: I have inherited this unit along with high voltage power supply(K1220VTS), freeze drying unit(K775) and the carbon coater(K250)attachment. The serial number for the K775 is 775-013. I would like to get a copy of manuals for all these units.
Is there any chance of obtaining a manual(s) from anyone?
thanks in advance
Fred Pearson
Fred Pearson Electron Optics Coordinator McMaster University Brockhouse Institute for Materials Research ABB-B145 1280 Main Street West Hamilton ON. Canada L8S 4M1 fax: (905) 521-2773 http://www.brockhouse.mcmaster.ca/
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Question: I am attempting to ultra-microtome polypropylene and high density polyethylene nanocomposites for examination of the clay dispersion with TEM. The samples are plastic pellets. Using a diamond knife I am able to easily prepare thick sections ~ 250 nm, however these are too thick to see the clay dispersions. When I cut in the range of ~ 90 nm it is difficult to get any sections and the sections I am able to obtain are quite wrinkled. Is there a chemical I can use to ëironí the sections? Should I embed the pellets in something instead of just directly sectioning the pellets? If so what? I have found papers that successfully use both room temperature and cryo-microtome; however I must do these at room temp.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gleasonkathryn-at-yahoo.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, October 24, 2006 at 10:25:41 ---------------------------------------------------------------------------
Email: gleasonkathryn-at-yahoo.com Name: Kathryn
Organization: NYC College of Technology
Education: Undergraduate College
Location: Brooklyn, NY, USA
Question: Can a person with liberal arts degrees but no science courses train to become a microscopist? If yes, what kind of institution provides training?
The short answer is "yes, but you're going to have to take some science classes", mostly physics, chemistry, and biology, if you're interesting in biological/biotech microscopy. Off the top of my head, I know of Central Michigan U. (obviously), Madison Area Technical College in Madison, WI, and San Joaquin Delta College in Stockton, CA. Other universities will have courses in various microscopies, but not so many have majors in microscopy like these 3.
Phil
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==============================Original Headers============================== 3, 23 -- From oshel1pe-at-cmich.edu Wed Oct 25 09:14:27 2006 3, 23 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9PEEQBm026398 3, 23 -- for {Microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 09:14:27 -0500 3, 23 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 23 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k9PEjCN6012466; 3, 23 -- Wed, 25 Oct 2006 10:45:15 -0400 3, 23 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 23 -- Wed, 25 Oct 2006 10:14:22 -0400 3, 23 -- Mime-Version: 1.0 3, 23 -- Message-Id: {f06230908c1651f92efea-at-[141.209.160.249]} 3, 23 -- In-Reply-To: {200610251245.k9PCjaBP011273-at-ns.microscopy.com} 3, 23 -- References: {200610251245.k9PCjaBP011273-at-ns.microscopy.com} 3, 23 -- Date: Wed, 25 Oct 2006 10:14:20 -0400 3, 23 -- To: gleasonkathryn-at-yahoo.com 3, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 23 -- Subject: Re: [Microscopy] AskAMicroscopist: train to become a microscopist? 3, 23 -- Cc: Microscopy-at-microscopy.com 3, 23 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 23 -- X-OriginalArrivalTime: 25 Oct 2006 14:14:22.0540 (UTC) FILETIME=[DE73D8C0:01C6F83F] 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==============================
Dear All, I have been having serious problems embedding cells grown on collagen gels. I have tried increasing my dehydration (EtOH) & infiltration steps. I have tried different resins (Spurr's and LX112). I still end up with gumdrops. These are not thin coatings of collagen as you would use to get finicky cells to grow on coverslips or dishes. These are actually cushions of collagen cast within the confines of cloning rings and probably about 30-50 micrometers thick when fully hydrated. Why should these be so much more problematic than tissue? The PI has also given me the same cells grown on fibronectin, and those are fine. Unfortunately, the cells behave a little differently on FN, and the structures the lab is studying aren't as abundant. HELP. thanks, 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, 22 -- From lcgould-at-med.cornell.edu Wed Oct 25 09:17:09 2006 1, 22 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9PEH8Os028417 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 09:17:08 -0500 1, 22 -- Received: from mpx3.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 22 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9PEEN3l013693 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 10:17:05 -0400 (EDT) 1, 22 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 22 -- by mpx3.med.cornell.edu 1, 22 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 22 -- with ESMTPA id {0J7P008HX2WNPD20-at-mpx3.med.cornell.edu} for 1, 22 -- microscopy-at-microscopy.com; Wed, 25 Oct 2006 10:00:24 -0400 (EDT) 1, 22 -- Date: Wed, 25 Oct 2006 10:00:21 -0400 1, 22 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 22 -- Subject: Microscopy: cells grown on collage gels 1, 22 -- Sender: lcgould-at-med.cornell.edu 1, 22 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 1, 22 -- Message-id: {p06230908c1651cf939b0-at-[140.251.48.23]} 1, 22 -- MIME-version: 1.0 1, 22 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.10.25.65933 ==============================End of - Headers==============================
There are (at least) 2 technical colleges which offer 2 year programs in electron microscopy (presumably that is what you refer to, not optical microscopy), Madison Area Technical College (here in Madison, WI), matcmadison.edu/electronmicros/ the other is San Joaquin Delta College in California www.deltacollege.edu/dept/electmicro/whatis.html I do not believe they require any science background per se.
John
-- ======================================================== John Fournelle, Ph.D. office: (608) 262-7964 cell: (608) 438-7480 Cameron Electron Microprobe Lab lab: (608) 265-4798 Dept of Geology & Geophysics fax: (608) 262-0693 University of Wisconsin home: (608) 274-2245 1215 West Dayton St. email: johnf-at-geology.wisc.edu Madison, WI 53706 amateur radio: WA3BTA Personal http://www.geology.wisc.edu/~johnf/ Probe lab http://www.geology.wisc.edu/~johnf/sx51.html Probe Sign Up Calender: http://www.microscopy.wisc.edu/cgi-bin/calendar/microprobe/calendar.cgi
"The first rule of all intelligent tinkering is to save every cog and wheel." -- Aldo Leopold
"For a successful technology, reality must take precedence over public relations, for Nature cannot be fooled." -- Richard P. Feynman
==============================Original Headers============================== 7, 24 -- From johnf-at-geology.wisc.edu Wed Oct 25 09:45:58 2006 7, 24 -- Received: from ice.geology.wisc.edu (ice.geology.wisc.edu [144.92.206.14]) 7, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9PEjw5V015714 7, 24 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 09:45:58 -0500 7, 24 -- Received: from localhost (localhost [127.0.0.1]) 7, 24 -- by localhost (Postfix) with ESMTP id A240320D4E 7, 24 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 09:45:57 -0500 (CDT) 7, 24 -- Received: from ice.geology.wisc.edu ([127.0.0.1]) 7, 24 -- by localhost (geology.wisc.edu [127.0.0.1]) (amavisd-new, port 10024) 7, 24 -- with ESMTP id 03952-02-2 for {microscopy-at-microscopy.com} ; 7, 24 -- Wed, 25 Oct 2006 09:43:50 -0500 (CDT) 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 6E68920D3E 7, 24 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 09:40:43 -0500 (CDT) 7, 24 -- Mime-Version: 1.0 7, 24 -- Message-Id: {p06230902c165260f08b7-at-[144.92.206.57]} 7, 24 -- Date: Wed, 25 Oct 2006 09:33:42 -0500 7, 24 -- To: microscopy-at-microscopy.com 7, 24 -- From: John Fournelle {johnf-at-geology.wisc.edu} 7, 24 -- Subject: Re: [Microscopy] AskAMicroscopist: train to become a microscopist? 7, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 7, 24 -- X-Virus-Scanned: amavisd-new at geology.wisc.edu ==============================End of - Headers==============================
Hi Kathryn, With respect to my fellow microscopist, I have to disagree. It would be like trying to become a race car driver without knowing how to drive.
No, you can not train to be a microscopist without a science background. A background in science is fundamental to any specific scientific field. You must acquire an understanding of how science should be conducted, how to learn outside your chosen field and then how to apply your knowledge to solve or accomplish a task. some of these skills you may have, others need to be developed. That were the science background is needed.
This is not to say you can not learn science while you learn microscopy. It's just a longer road.
Best wishes and good luck with your studies.
Frank Karl
gleasonkathryn-at-ya hoo.com To: frank.karl-at-degussa.com cc: 10/25/2006 07:39 Subject: [Microscopy] AskAMicroscopist: train to become a microscopist? AM Please respond to gleasonkathryn
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Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gleasonkathryn-at-yahoo.com) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, October 24, 2006 at 10:25:41 ---------------------------------------------------------------------------
Email: gleasonkathryn-at-yahoo.com Name: Kathryn
Organization: NYC College of Technology
Education: Undergraduate College
Location: Brooklyn, NY, USA
Question: Can a person with liberal arts degrees but no science courses train to become a microscopist? If yes, what kind of institution provides training?
I have had some success with Collagen gel in the TEM.
Tips: Small size, and dehydrate. Days and Days... lots of changes... collagen likes water, and that's where we've had the problem both in the CPD and with the Spurr's.
We found tt was impossible to dehydrate the collagen while it was attached to a coverslip, it held onto the water very strongly.
I don't know if this helps, but other than doing many many 100% ethanol changes prior to infiltration with the resin the samples weren't treated any differently and we had decent results.
HTH
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: lcgould-at-med.cornell.edu [mailto:lcgould-at-med.cornell.edu] Sent: Wednesday, October 25, 2006 10:22 AM To: Williams, Geoffrey
Dear All, I have been having serious problems embedding cells grown on collagen gels. I have tried increasing my dehydration (EtOH) & infiltration steps. I have tried different resins (Spurr's and LX112). I still end up with gumdrops. These are not thin coatings of collagen as you would use to get finicky cells to grow on coverslips or dishes. These are actually cushions of collagen cast within the confines of cloning rings and probably about 30-50 micrometers thick when fully hydrated. Why should these be so much more problematic than tissue? The PI has also given me the same cells grown on fibronectin, and those are fine. Unfortunately, the cells behave a little differently on FN, and the structures the lab is studying aren't as abundant. HELP. thanks, 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, 22 -- From lcgould-at-med.cornell.edu Wed Oct 25 09:17:09 2006 1, 22 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9PEH8Os028417 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 09:17:08 -0500 1, 22 -- Received: from mpx3.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 22 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9PEEN3l013693 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 10:17:05 -0400 (EDT) 1, 22 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 22 -- by mpx3.med.cornell.edu 1, 22 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 22 -- with ESMTPA id {0J7P008HX2WNPD20-at-mpx3.med.cornell.edu} for 1, 22 -- microscopy-at-microscopy.com; Wed, 25 Oct 2006 10:00:24 -0400 (EDT) 1, 22 -- Date: Wed, 25 Oct 2006 10:00:21 -0400 1, 22 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 1, 22 -- Subject: Microscopy: cells grown on collage gels 1, 22 -- Sender: lcgould-at-med.cornell.edu 1, 22 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 1, 22 -- Message-id: {p06230908c1651cf939b0-at-[140.251.48.23]} 1, 22 -- MIME-version: 1.0 1, 22 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 22 -- Content-transfer-encoding: 7BIT 1, 22 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.10.25.65933 ==============================End of - Headers==============================
==============================Original Headers============================== 13, 30 -- From Geoffrey_Williams-at-brown.edu Wed Oct 25 12:42:55 2006 13, 30 -- Received: from perseus.services.brown.edu (perseus.services.brown.edu [128.148.106.173]) 13, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9PHgtX0007988 13, 30 -- for {Microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 12:42:55 -0500 13, 30 -- Received: from ex-gateway1-out.AD.Brown.Edu (ex-gateway1.ad.brown.edu [128.148.21.51]) 13, 30 -- by perseus.services.brown.edu (Switch-3.1.10/Switch-3.1.7/) with ESMTP id k9PHgppI007231; 13, 30 -- Wed, 25 Oct 2006 13:42:51 -0400 (EDT) 13, 30 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex-gateway1-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); 13, 30 -- Wed, 25 Oct 2006 13:42:51 -0400 13, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 13, 30 -- Content-class: urn:content-classes:message 13, 30 -- MIME-Version: 1.0 13, 30 -- Content-Type: text/plain; 13, 30 -- charset="us-ascii" 13, 30 -- Subject: RE: [Microscopy] Microscopy: cells grown on collage gels 13, 30 -- Date: Wed, 25 Oct 2006 13:42:51 -0400 13, 30 -- Message-ID: {A1A84D541C161C4C988B4E0FB38F5A0F04B90E68-at-MAIL1.AD.Brown.Edu} 13, 30 -- In-Reply-To: {200610251421.k9PELgKW007065-at-ns.microscopy.com} 13, 30 -- X-MS-Has-Attach: 13, 30 -- X-MS-TNEF-Correlator: 13, 30 -- Thread-Topic: [Microscopy] Microscopy: cells grown on collage gels 13, 30 -- Thread-Index: Acb4QOZ+49xnNFNCSiWfjhsbL+fsngAG1sqQ 13, 30 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} 13, 30 -- To: {lcgould-at-med.cornell.edu} , {Microscopy-at-microscopy.com} 13, 30 -- X-OriginalArrivalTime: 25 Oct 2006 17:42:51.0720 (UTC) FILETIME=[FE813C80:01C6F85C] 13, 30 -- X-Brown-Proofpoint: Not Infected 13, 30 -- X-Proofpoint-Spam-Details: rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0609290000 definitions=main-0610250020 13, 30 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe engine=3.1.0-0609290000 definitions=main-0610250020 13, 30 -- Content-Transfer-Encoding: 8bit 13, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k9PHgtX0007988 ==============================End of - Headers==============================
If water is the problem, you might try one of the plastics that do not mind some water. David
On Oct 25, 2006, at 10:46 AM, 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 } ---------------------------------------------------------------------- } ------ } } I have had some success with Collagen gel in the TEM. } } Tips: } Small size, and dehydrate. Days and Days... lots of changes... } collagen likes water, and that's where we've had the problem both } in the } CPD and with the Spurr's. } } We found tt was impossible to dehydrate the collagen while it was } attached to a coverslip, it held onto the water very strongly. } } I don't know if this helps, but other than doing many many 100% } ethanol } changes prior to infiltration with the resin the samples weren't } treated } any differently and we had decent results. } } HTH } } Geoff Williams } Leduc Bioimaging Facility Manager } Brown University } } http://www.brown.edu/Facilities/Leduc_Bioimaging_Facility/ } } -----Original Message----- } X-from: lcgould-at-med.cornell.edu [mailto:lcgould-at-med.cornell.edu] } Sent: Wednesday, October 25, 2006 10:22 AM } To: Williams, Geoffrey } Subject: [Microscopy] Microscopy: cells grown on collage gels } } } } } ---------------------------------------------------------------------- } -- } ---- } 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 been having serious problems embedding cells grown on collagen } gels. I have tried increasing my dehydration (EtOH) & infiltration } steps. I have tried different resins (Spurr's and LX112). I still } end up with gumdrops. } These are not thin coatings of collagen as you would use to get } finicky cells to grow on coverslips or dishes. These are actually } cushions of collagen cast within the confines of cloning rings and } probably about 30-50 micrometers thick when fully hydrated. } Why should these be so much more problematic than tissue? } The PI has also given me the same cells grown on fibronectin, and } those are fine. Unfortunately, the cells behave a little differently } on FN, and the structures the lab is studying aren't as abundant. } HELP. } thanks, } 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, 22 -- From lcgould-at-med.cornell.edu Wed Oct 25 09:17:09 2006 } 1, 22 -- Received: from smtp-gw2.med.cornell.edu } (smtp-gw2.med.cornell.edu [157.139.3.45]) } 1, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k9PEH8Os028417 } 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 } 09:17:08 -0500 } 1, 22 -- Received: from mpx3.med.cornell.edu } (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) } 1, 22 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) } with ESMTP id k9PEEN3l013693 } 1, 22 -- for {microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 } 10:17:05 -0400 (EDT) } 1, 22 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu } [140.251.48.23]) } 1, 22 -- by mpx3.med.cornell.edu } 1, 22 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan } 28 2005)) } 1, 22 -- with ESMTPA id {0J7P008HX2WNPD20-at-mpx3.med.cornell.edu} for } 1, 22 -- microscopy-at-microscopy.com; Wed, 25 Oct 2006 10:00:24 -0400 } (EDT) } 1, 22 -- Date: Wed, 25 Oct 2006 10:00:21 -0400 } 1, 22 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} } 1, 22 -- Subject: Microscopy: cells grown on collage gels } 1, 22 -- Sender: lcgould-at-med.cornell.edu } 1, 22 -- To: Microscopy Listserver {microscopy-at-microscopy.com} } 1, 22 -- Message-id: {p06230908c1651cf939b0-at-[140.251.48.23]} } 1, 22 -- MIME-version: 1.0 } 1, 22 -- Content-type: text/plain; charset=us-ascii; format=flowed } 1, 22 -- Content-transfer-encoding: 7BIT } 1, 22 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, } Antispam-Data: 2006.10.25.65933 } ==============================End of - } Headers============================== } } } ==============================Original } Headers============================== } 13, 30 -- From Geoffrey_Williams-at-brown.edu Wed Oct 25 12:42:55 2006 } 13, 30 -- Received: from perseus.services.brown.edu } (perseus.services.brown.edu [128.148.106.173]) } 13, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k9PHgtX0007988 } 13, 30 -- for {Microscopy-at-microscopy.com} ; Wed, 25 Oct 2006 } 12:42:55 -0500 } 13, 30 -- Received: from ex-gateway1-out.AD.Brown.Edu (ex- } gateway1.ad.brown.edu [128.148.21.51]) } 13, 30 -- by perseus.services.brown.edu (Switch-3.1.10/ } Switch-3.1.7/) with ESMTP id k9PHgppI007231; } 13, 30 -- Wed, 25 Oct 2006 13:42:51 -0400 (EDT) } 13, 30 -- Received: from mail1.AD.Brown.Edu ([128.148.21.30]) by ex- } gateway1-out.AD.Brown.Edu with Microsoft SMTPSVC(6.0.3790.1830); } 13, 30 -- Wed, 25 Oct 2006 13:42:51 -0400 } 13, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 13, 30 -- Content-class: urn:content-classes:message } 13, 30 -- MIME-Version: 1.0 } 13, 30 -- Content-Type: text/plain; } 13, 30 -- charset="us-ascii" } 13, 30 -- Subject: RE: [Microscopy] Microscopy: cells grown on } collage gels } 13, 30 -- Date: Wed, 25 Oct 2006 13:42:51 -0400 } 13, 30 -- Message-ID: } {A1A84D541C161C4C988B4E0FB38F5A0F04B90E68-at-MAIL1.AD.Brown.Edu} } 13, 30 -- In-Reply-To: {200610251421.k9PELgKW007065-at-ns.microscopy.com} } 13, 30 -- X-MS-Has-Attach: } 13, 30 -- X-MS-TNEF-Correlator: } 13, 30 -- Thread-Topic: [Microscopy] Microscopy: cells grown on } collage gels } 13, 30 -- Thread-Index: Acb4QOZ+49xnNFNCSiWfjhsbL+fsngAG1sqQ } 13, 30 -- From: "Williams, Geoffrey" {Geoffrey_Williams-at-brown.edu} } 13, 30 -- To: {lcgould-at-med.cornell.edu} , {Microscopy-at-microscopy.com} } 13, 30 -- X-OriginalArrivalTime: 25 Oct 2006 17:42:51.0720 (UTC) } FILETIME=[FE813C80:01C6F85C] } 13, 30 -- X-Brown-Proofpoint: Not Infected } 13, 30 -- X-Proofpoint-Spam-Details: rule=notspam policy=default } score=0 mlx=-1 adultscore=0 adjust=0 reason=safe } engine=3.1.0-0609290000 definitions=main-0610250020 } 13, 30 -- X-Brown-MailScanner-SpamCheck: not spam, rule=notspam } policy=default score=0 mlx=-1 adultscore=0 adjust=0 reason=safe } engine=3.1.0-0609290000 definitions=main-0610250020 } 13, 30 -- Content-Transfer-Encoding: 8bit } 13, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k9PHgtX0007988 } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-arizona.edu Wed Oct 25 19:35:06 2006 5, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9Q0Z6d0027035 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 25 Oct 2006 19:35:06 -0500 5, 22 -- Received: from localhost (eomer.email.arizona.edu [10.0.0.219]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id C8E48118C805 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 25 Oct 2006 17:35:05 -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 810D311AD7AB 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 25 Oct 2006 17:34:49 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 22 -- In-Reply-To: {200610251746.k9PHkTEn013403-at-ns.microscopy.com} 5, 22 -- References: {200610251746.k9PHkTEn013403-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {1789A975-87FF-48F7-9D3B-D43D8E9F821C-at-arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-arizona.edu} 5, 22 -- Subject: Re: [Microscopy] RE: Microscopy: cells grown on collage gels 5, 22 -- Date: Wed, 25 Oct 2006 17:34:48 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.752.2) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
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Email: ionsourcerer-at-mac.com Name: Rick Becker
Organization: Cluster Sciences, a.k.a Ibadex Research
Title-Subject: [Filtered] Philips EM 301 TEM assembly/service manual
Question: Might anyone out there have a manual I could purchase or copy?
Some wonderful folks at Harvard are bequeathing the machine to me if I can make it go away. To do so requires complete dissassembly of the column, (and everything else) into countless small boxes for transport in my little pick-up truck. I plan to mark, tape, label, and photograph everything to the best of my ability, but it will need to be in storage until I move to a new lab space, so I'll need every available crutch when it's time to put it back together. All of the other documentation is complete, but a good set of mechanical drawings would be priceless. Neighborly advice is _always_ appreciated.
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Email: squinto-at-natural-immunogenics.com Name: Stephen Quinto
Organization: Natural-Immunogenics Corp.
Title-Subject: [Filtered] Position Available
Question: Materials Specialist - TEM Microscopist - [physics, molecular biology, electro-chemistry or similar background - college level education] is sought by our company [located in Pompano Beach Florida].
The laboratory at Natural-Immunogenics Corp {http://www.natural-immunogenics.com/} www.natural-immunogenics.com is responsible for R&D, analytical work and Quality Control. A review of our website will give anyone interested a broader understanding of the prospects than one can put into a short message. Further, an even deeper appreciation of the clinical potential of these 'products' may be had by visiting {http://www.imref.org/} www.imref.org - a foundation established and endowed by Natural-Immunogenics for its own stated purposes.
Our company has a Phillips EM-400T but continues to investigate the possibility of upgrading that capability. It is also increasing the members of its team to respond to the growing demand upon its expertise, among which is also a joint venture in Asia. We have just produced a gold hydrosol that has resulted in near-perfect dispersion of gold particles close to five Angstroms in size - probably an unprecedented accomplishment.
Interested parties must be experienced and self-reliant as whomsoever is chosen will have to learn quickly to replace the 'authority' on pico-scalar mineral hydrosols. So at least several years of experience is desired. Our microscopist will need both to operate and supervise maintenance of the equipment. As we are a small company [only twenty-four of us] other duties in the lab and production will also be required.
Thank you all for your hints, tips and insight. The general consensus is that collagen gels are just mighty sponges that require very rigorous and prolonged dehydrations. Anything that helps accomplish that is good: releasing the gels from the dish, cutting them into smaller pieces, en bloc staining with UA, using multiple changes of PO before the resin, using a resin with more "tolerance" for a little water. The PI was concerned that the integrity of the cell monolayer would be compromised by removing the gels from the dish and/or cutting them into pieces. I think that I've convinced him that slight damage to a monolayer that I can then cut and we can look at is VASTLY preferable to what we have at the moment.
Thank you again. As always, you guys are a font of information. 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, 22 -- From lcgould-at-med.cornell.edu Thu Oct 26 08:43:40 2006 2, 22 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 2, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9QDheXQ009680 2, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 26 Oct 2006 08:43:40 -0500 2, 22 -- Received: from mpx3.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 2, 22 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k9QDhNhY028523 2, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 26 Oct 2006 09:43:37 -0400 (EDT) 2, 22 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 2, 22 -- by mpx3.med.cornell.edu 2, 22 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 2, 22 -- with ESMTPA id {0J7Q0082AWSL9720-at-mpx3.med.cornell.edu} for 2, 22 -- microscopy-at-msa.microscopy.com; Thu, 26 Oct 2006 09:43:34 -0400 (EDT) 2, 22 -- Date: Thu, 26 Oct 2006 09:43:30 -0400 2, 22 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 2, 22 -- Subject: cells on collagen gels 2, 22 -- Sender: lcgould-at-med.cornell.edu 2, 22 -- To: "microscopy-at-msa.microscopy.com" {microscopy-at-msa.microscopy.com} 2, 22 -- Message-id: {p06230906c1666a876729-at-[140.251.48.23]} 2, 22 -- MIME-version: 1.0 2, 22 -- Content-type: text/plain; charset=us-ascii; format=flowed 2, 22 -- Content-transfer-encoding: 7BIT 2, 22 -- X-PMX-Version: 5.2.0.264296, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.10.26.62934 ==============================End of - Headers==============================
I have often used streptavidin conjugated fluorochromes but now find myself in need of a biotin-conjugated fluorochrome (e.g., FITC, Alexa) for a special experiment. Does anyone know a commercial source?
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
I am interested in purchasing a Tecscan Vega II-LMU SEM. Any feedback on or off list is appreciated.
-- Gerald Bourne, Ph.D. Major Analytical Instrumentation Center Department of Materials Science and Engineering University of Florida 107H MAEC P.O. Box 116400 Gainesville, FL 32611 (352) 392-6985 (352) 392-0390 fax
==============================Original Headers============================== 4, 24 -- From grb-at-ufl.edu Thu Oct 26 16:57:49 2006 4, 24 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9QLvm7Q007617 4, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Oct 2006 16:57:49 -0500 4, 24 -- Received: from [10.245.16.5] ([10.245.16.5]) 4, 24 -- (authenticated bits=0) 4, 24 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id k9QLvkYB5353658 4, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 4, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 26 Oct 2006 17:57:47 -0400 4, 24 -- Message-ID: {45412F79.2070107-at-ufl.edu} 4, 24 -- Date: Thu, 26 Oct 2006 17:58:17 -0400 4, 24 -- From: Gerald Bourne {grb-at-ufl.edu} 4, 24 -- User-Agent: Mozilla Thunderbird 1.0.7 (Windows/20050923) 4, 24 -- X-Accept-Language: en-us, en 4, 24 -- MIME-Version: 1.0 4, 24 -- To: Microscopy-at-microscopy.com 4, 24 -- Subject: Tecscan SEM 4, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 24 -- Content-Transfer-Encoding: 7bit 4, 24 -- X-Greylist: Sender succeeded SMTP AUTH authentication, not delayed by milter-greylist-3.0rc3 (smtp.ufl.edu [128.227.74.56]); Thu, 26 Oct 2006 17:57:47 -0400 (EDT) 4, 24 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 24 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 4, 24 -- X-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) 4, 24 -- X-UFL-Scanned-By: CNS Open Systems Group (http://open-systems.ufl.edu/services/smtp-relay/) ==============================End of - Headers==============================
We have a investigator who would like to image an oil-in-water nanoemulsion. We have tried SEM and also applying it directly to carbon-coated grids, fixing with osmium, and viewing by TEM. The problem is that the nanoemulsion coalesces during processing. What would be the best technique to process and image this type of sample?
Thanks for your suggestions.
Dotty Sorenson
Dorothy Sorenson Microscopy and Image-analysis Laboratory Department of Cell and Developmental Biology University Of Michigan Medical School A807 BSRB 109 Zina Pitcher Place Ann Arbor, MI 48109-2200 (734)763-1170 FAX (734)763-1166
==============================Original Headers============================== 7, 16 -- From dsoren-at-umich.edu Fri Oct 27 08:29:10 2006 7, 16 -- Received: from hellskitchen.mr.itd.umich.edu (smtp.mail.umich.edu [141.211.14.82]) 7, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9RDTAi1002445 7, 16 -- for {microscopy-at-msa.microscopy.com} ; Fri, 27 Oct 2006 08:29:10 -0500 7, 16 -- Received: FROM [10.21.129.138] (host-18.subnet-17.med.umich.edu [141.214.17.18]) 7, 16 -- BY hellskitchen.mr.itd.umich.edu ID 45420969.EF6A2.735 ; 7, 16 -- 27 Oct 2006 09:28:10 -0400 7, 16 -- Mime-Version: 1.0 (Apple Message framework v752.2) 7, 16 -- Content-Transfer-Encoding: 7bit 7, 16 -- Message-Id: {AB1C4513-348E-4429-A697-BC66675E83CB-at-umich.edu} 7, 16 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 16 -- To: microscopy-at-msa.microscopy.com 7, 16 -- From: Dorothy Sorenson {dsoren-at-umich.edu} 7, 16 -- Subject: EM nanoemulsion 7, 16 -- Date: Fri, 27 Oct 2006 09:25:23 -0400 7, 16 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
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Email: mgb-at-ansto.gov.au Name: BLACKFORD, Mark
Organization: ANSTO
Title-Subject: [Filtered] Positions Open Professional Officer Electron Microscopy and Research Scientist Transmission Electron Microscopy
Question: Organisation: Institute of Materials and Engineering Science Australian Nuclear Science and Technology Organisation (ANSTO)
We are seeking to fill two new electron microscopy positions in the Materials Characterisation Section at ANSTO. Please see full descriptions at the web addresses below:
1) Professional Officer (Electron Microscopy) http://www.ansto.gov.au/info/vac/vac2006/anstovacancy2006_126.pdf
2) Research Scientist (Transmission Electron Microscopy) http://www.ansto.gov.au/info/vac/vac2006/anstovacancy2006_125.pdf
Thank You,
Mark Blackford Institute of Materials and Engineering Science ANSTO PMB 1, Menai, N.S.W., 2234 Australia
The best way to do this is cryoEM. Place a sample on e.g. a TEM grid, plunge into slush nitrogen (almost frozen LN2), place on a cryostage, cryo-sputter coat, and into a low-voltage FESEM. The Philips XL30ESEM at the U Mich North Campus EMAL lists a cryostage, so you might want to contact them to see if they can do your sample. CryoTEM might also work, but I've only done cryoSEM. Any method other than cryo can easily distort the emulsion droplets, and give inaccurate results.
Phil P.S. Sorry, we don't have cryoEM capability here at CMU, or I'd invite you up.
} Hello all, } } We have a investigator who would like to image an oil-in-water } nanoemulsion. We have tried SEM and also applying it directly to } carbon-coated grids, fixing with osmium, and viewing by TEM. The } problem is that the nanoemulsion coalesces during processing. What } would be the best technique to process and image this type of sample? } } Thanks for your suggestions. } } Dotty Sorenson } } Dorothy Sorenson } Microscopy and Image-analysis Laboratory } Department of Cell and Developmental Biology } University Of Michigan Medical School } A807 BSRB } 109 Zina Pitcher Place } Ann Arbor, MI 48109-2200 } (734)763-1170 } FAX (734)763-1166 -- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 4, 22 -- From oshel1pe-at-cmich.edu Fri Oct 27 09:04:56 2006 4, 22 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9RE4ubJ024373 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 27 Oct 2006 09:04:56 -0500 4, 22 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 22 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k9REZfN2014423 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 27 Oct 2006 10:35:43 -0400 4, 22 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 22 -- Fri, 27 Oct 2006 10:04:54 -0400 4, 22 -- Mime-Version: 1.0 4, 22 -- Message-Id: {f06230909c167c03c7065-at-[141.209.160.249]} 4, 22 -- In-Reply-To: {200610271335.k9RDZpsD009758-at-ns.microscopy.com} 4, 22 -- References: {200610271335.k9RDZpsD009758-at-ns.microscopy.com} 4, 22 -- Date: Fri, 27 Oct 2006 10:04:52 -0400 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 22 -- Subject: Re: [Microscopy] EM nanoemulsion 4, 22 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 22 -- X-OriginalArrivalTime: 27 Oct 2006 14:04:54.0657 (UTC) FILETIME=[E0CB4710:01C6F9D0] 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, When I worked at a rubber company, we looked synthetic latex by treating with osmium and cryomicrotoming. A small plastic capillary tube held the treated sample in the microtome. this was frozen in the cryomicrotome. Thin sections were cut with a diamond knife and floated on icy cold 50/50 DSMO and water. The thin sections were picked up with a "perfect loop" and transfer to a carbon film grid. Of course the latex spheres were stable after hardening, I don't know about your oil.
We were looking for hollow latex spheres,and we found a few. I was never convinced these rings were real and not some cutting artifact which dropped a section of core out of the hardened skin.
Good luck, let me know how it works out.
Frank Karl Degussa Corporation Akron Technical Center 3500 Embassy Parkway Suite 100 Akron, Ohio 44333 330-668-2235 Ext. 238 This e-mail transmission, and any documents, files or previous e-mail messages attached to it may contain information that is confidential or legally privileged. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that you must not read this transmission and that any disclosure, copying, printing, distribution or use of any of the information contained in or attached to this transmission is STRICTLY PROHIBITED. If you have received this transmission in error, please immediately notify the sender by telephone or return e-mail and delete the original transmission and its attachments without reading or saving in any manner. Thank you.
==============================Original Headers============================== 6, 18 -- From frank.karl-at-degussa.com Fri Oct 27 15:07:41 2006 6, 18 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9RK7eGv013904 6, 18 -- for {microscopy-at-msa.microscopy.com} ; Fri, 27 Oct 2006 15:07:40 -0500 6, 18 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 6, 18 -- by framailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id k9RK7aqE001634; 6, 18 -- Fri, 27 Oct 2006 22:07:37 +0200 6, 18 -- In-Reply-To: {200610271331.k9RDVph1004635-at-ns.microscopy.com} 6, 18 -- Subject: Re: [Microscopy] EM nanoemulsion 6, 18 -- To: dsoren-at-umich.edu, microscopy-at-msa.microscopy.com 6, 18 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 6, 18 -- Message-ID: {OFC55AE551.E3B839E5-ON86257214.006D97A5-86257214.006E871F-at-degussa.com} 6, 18 -- From: frank.karl-at-degussa.com 6, 18 -- Date: Fri, 27 Oct 2006 15:07:16 -0500 6, 18 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 6, 18 -- 10/27/2006 03:07:38 PM 6, 18 -- MIME-Version: 1.0 6, 18 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
I read your request for manuals on the various Emitech Instruments, and have requested manuals for each piece of equipment to be sent directly to you. For your reference, Quorum Technologies within the last year, purchased Emitech, and now handles both Emitech and the Polaron line of EM preparation equipment. Energy Beam Sciences is the master distributor here in the US, with Soquelec Ltd distributing equipment in Canada.(514) 482-6427.
If you have any further questions, or require additional information please contact us.
Mike Dufraine Energy Beam Sciences,Inc.
eoptics-at-mcmaster.ca wrote:
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-- Michael F. Dufraine EM - Product Manager TEL 800-992-9037 X340 MDufraine-at-ebsciences.com www.ebsciences.com
==============================Original Headers============================== 10, 26 -- From mdufraine-at-ebsciences.com Fri Oct 27 15:45:30 2006 10, 26 -- Received: from nlpi043.sbcis.sbc.com (nlpi043.sbcis.sbc.com [207.115.36.72]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9RKjU0M025323 10, 26 -- for {microscopy-at-microscopy.com} ; Fri, 27 Oct 2006 15:45:30 -0500 10, 26 -- X-ORBL: [69.182.224.2] 10, 26 -- Received: from mail.ebsciences.com ([69.182.224.2]) 10, 26 -- by nlpi043.sbcis.sbc.com (8.13.8 out.dk.spool/8.13.7) with ESMTP id k9RKj7b4006850; 10, 26 -- Fri, 27 Oct 2006 15:45:07 -0500 10, 26 -- Received: from mdufraine.ebsciences.private ([10.10.0.196]) 10, 26 -- by mail.ebsciences.com with esmtpsa (TLSv1:AES256-SHA:256) 10, 26 -- (Exim 4.62) 10, 26 -- (envelope-from {mdufraine-at-ebsciences.com} ) 10, 26 -- id 1GdYZs-00052N-CP; Fri, 27 Oct 2006 16:45:20 -0400 10, 26 -- Message-ID: {45426FDF.1090201-at-ebsciences.com} 10, 26 -- Date: Fri, 27 Oct 2006 16:45:19 -0400 10, 26 -- From: Mike Dufraine {mdufraine-at-ebsciences.com} 10, 26 -- Organization: Energy Beam Sciences 10, 26 -- User-Agent: Mozilla Thunderbird 1.0.2 (Windows/20050317) 10, 26 -- X-Accept-Language: en-us, en 10, 26 -- MIME-Version: 1.0 10, 26 -- To: eoptics-at-mcmaster.ca, microscopy-at-microscopy.com 10, 26 -- Subject: Re: [Microscopy] viaWWW: Emitech K775 and K250 Manuals 10, 26 -- References: {200610251237.k9PCbSDY018269-at-ns.microscopy.com} 10, 26 -- In-Reply-To: {200610251237.k9PCbSDY018269-at-ns.microscopy.com} 10, 26 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 26 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
MSD is currently searching for an Electron Microscopist to provide support to the National Institutes of Health (NIH) scientific staff. This opportunity is a permanent, full-time position with MSD and it is on-site at the NIH's Rocky Mountain Laboratories in Hamilton, Montana. MSD is an employee-owned company of over 600 professionals who support the diverse scientific, technical, and administrative needs of the Federal government.
Major Duties and Responsibilities: * Complete preparation and imaging of biological specimens for conventional, cryo-, and immuno- SEM and TEM. * Perform TEM sample preparation that includes appropriate fixation, embedding, sectioning and staining of animal tissues and cell culture samples. * Perform SEM sample preparation that includes appropriate fixation, dehydration, critical point drying, and sputter coating. * Perform cryo preparative techniques that include use of a high pressure freezer, freeze substitution equipment, cryo-ultramicrotomy, plunge freezing, and use of cryo-transfer devices and microscope stages. * Perform single or multiple immunolabeling that involves both pre- and post embedding techniques with labeling of unfixed or lightly fixed specimens, resin sections, cryo sections (Tokuyasu), and/or immunocytochemistry. * Additional responsibilities include participation in group meetings to discuss projects, sample and image preparation, data collection, and assistance in micrograph interpretation for scientific staff and database archive.
Position Requirements: * B.A./B.S. in Microbiology, Biology, Biochemistry, or related field required. M.S. or Ph.D. is preferred. * Minimum of two years electron microscopy experience. * Experience with both TEM and SEM. * Experience with conventional, cryo-, and immunolabeling techniques. * Microscopy experience should include sample preparation and image collection. * Experience with a variety of tissue types is preferred. * Expertise in pathology, histology, or biomedical ultrastructural analysis is preferred. * History of research publications preferred. * Excellent communication skills and ability to work in a team environment with all levels of co-workers are essential.
Please send CV and salary requirements to: David Dowling Sr. Scientific Recruiter MSD, Inc. ddowling-at-msdinc.com
------------------------------------- Joseph L. Tasto, MD Director, Biomedical Affairs MSD, Inc. 2677 Prosperity Ave, Suite 700 Fairfax, VA 22031 www.msdinc.com
==============================Original Headers============================== 8, 20 -- From JTasto-at-msdinc.com Fri Oct 27 16:39:26 2006 8, 20 -- Received: from fairfax.msdinc.com (fairfax.msdinc.com [216.88.62.245]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9RLdQUH004356 8, 20 -- for {Microscopy-at-Microscopy.Com} ; Fri, 27 Oct 2006 16:39:26 -0500 8, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 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: Position Announcement: Electron Microscopist 8, 20 -- Date: Fri, 27 Oct 2006 17:39:30 -0400 8, 20 -- Message-ID: {D7B87A42EA865D49907BBB73848A47A502DF18-at-fairfax.inside.msdinc.com} 8, 20 -- X-MS-Has-Attach: 8, 20 -- X-MS-TNEF-Correlator: 8, 20 -- Thread-Topic: Position Announcement: Electron Microscopist 8, 20 -- Thread-Index: Acb6EGJVLUfd9NN1QgOXCc9XZzfFhQ== 8, 20 -- From: "Joseph L. Tasto, MD" {JTasto-at-msdinc.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 k9RLdQUH004356 ==============================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 pbgrover-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: pbgrover-at-yahoo.com Name: Paul Grover
Organization: Purdue University
Title-Subject: [Filtered] new forum on research ethics
Question: Hello fellow microscopists,
Back in April we had a thread relating to ethical practices in performing and reporting research. I had felt virtually alone in my concerns until I saw all the responses. I am thankful to all who replied, both on- and offlist. So by popular demand I have, with rapidity characteristic of a thundering herd of turtles, finally gotten around to setting up a Yahoo Group called Integrity in Scientific Research:
http://tech.groups.yahoo.com/group/honestscience/
If you are interested in this topic, please join and contribute. Also please alert others you think would be interested, and let me know any suggestions you have for the forum, and where I can advertise for new members.
I would like to know how to prepare cross section samples of multilayers on stainless steel substrate. I have the basic knowledge on the sample preparation of layers on silicon wafer using M-bond and ion thinning. However, stainless steel will be difficult to cut and grind. My sample is multilayers (such as Al2O3) on stainless steel substrate, and the thickness of multi-layers is around 3 um. Each layer has several 10 to 1 um in thickness. The other side of the multilayers is also stainless steel. The multi-layers are sandwiched by stainless steel. I will have 5 X 5 X 5 mm cubes. I will try to make TEM samples using ion-thining, and later try to use FIB. I would be appreciate if you could provide me any suggestions.
Thank you,
Hiromi Konishi, Ph.D. Laboratory Manager The S.W. Bailey X-ray Diffraction Laboratory Room A353 Weeks Hall Voice: (608) 262-9784, (608) 262-0915 Fax: (608) 262-0693 Department of Geology and Geophysics University of Wisconsin-Madison 1215 W Dayton St. Madison WI 53706
==============================Original Headers============================== 5, 30 -- From hikonishi-at-gmail.com Mon Oct 30 11:12:44 2006 5, 30 -- Received: from nz-out-0102.google.com (nz-out-0102.google.com [64.233.162.196]) 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9UHCiwK029303 5, 30 -- for {Microscopy-at-microscopy.com} ; Mon, 30 Oct 2006 11:12:44 -0600 5, 30 -- Received: by nz-out-0102.google.com with SMTP id r28so1227347nza 5, 30 -- for {Microscopy-at-microscopy.com} ; Mon, 30 Oct 2006 09:12:43 -0800 (PST) 5, 30 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 30 -- s=beta; d=gmail.com; 5, 30 -- h=received:message-id:from:to:subject:date:mime-version:content-type:content-transfer-encoding:x-priority:x-msmail-priority:x-mailer:x-mimeole; 5, 30 -- b=Jldmol8W9oGLiMNFyz0UELzSB1lm5/VlowaOg3Y0nHr0I7KnVF8GbMlMD29ktnvvbjJSre5bWx+j/e+9vY98gl/MYmxBaC4hjFeRf8JAKkyEy39VCZLSop1NN365FHSLItPUzoUYkdL+Mzrpg+wiiebRPrtaQBMN2772y2WoKyE= 5, 30 -- Received: by 10.35.51.19 with SMTP id d19mr5316975pyk; 5, 30 -- Mon, 30 Oct 2006 09:12:43 -0800 (PST) 5, 30 -- Received: from HKONISHI ( [144.92.207.106]) 5, 30 -- by mx.google.com with ESMTP id 19sm12937602nzp.2006.10.30.09.12.41; 5, 30 -- Mon, 30 Oct 2006 09:12:42 -0800 (PST) 5, 30 -- Message-ID: {003201c6fc46$a2504e40$6acf5c90-at-HKONISHI} 5, 30 -- From: "Hiromi Konishi" {hikonishi-at-gmail.com} 5, 30 -- To: {Microscopy-at-microscopy.com} 5, 30 -- Subject: cross section of multilayers on stainless substrate 5, 30 -- Date: Mon, 30 Oct 2006 11:12:47 -0600 5, 30 -- MIME-Version: 1.0 5, 30 -- Content-Type: text/plain; 5, 30 -- format=flowed; 5, 30 -- charset="iso-2022-jp"; 5, 30 -- reply-type=original 5, 30 -- Content-Transfer-Encoding: 7bit 5, 30 -- X-Priority: 3 5, 30 -- X-MSMail-Priority: Normal 5, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 5, 30 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
We need to upgrade our SEM capabilities to field emission. Most of the imaging would involve nanomaterials (spheres, wires, tubes, etc.) as well as biological macromolecules formed on silicon substrates. Most materials are nonconducting and in need of resolution in the 1-3 nm range. Regarding size of specimens: 3-4 mm to 4-5 inches. In addition, we would be adding ED X-ray microanalytic capability.
I would appreciate input/opinions from users of various brands of field emission SEMs in terms of: reliability (downtime), service support, cost of service contracts, software issues (problems).
Thank you.
John B. -- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
==============================Original Headers============================== 4, 18 -- From bozzola-at-siu.edu Mon Oct 30 12:07:15 2006 4, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9UI7F8m008397 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Mon, 30 Oct 2006 12:07:15 -0600 4, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 4, 18 -- by abbmta2.siu.edu (Switch-3.1.11/Switch-3.1.10) with ESMTP id k9UI7Diu015693 4, 18 -- for {Microscopy-at-msa.microscopy.com} ; Mon, 30 Oct 2006 12:07:14 -0600 (CST) 4, 18 -- Mime-Version: 1.0 4, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 4, 18 -- Message-Id: {p0611042bc16bdd0f8a67-at-[131.230.177.142]} 4, 18 -- Date: Mon, 30 Oct 2006 12:07:10 -0500 4, 18 -- To: Microscopy-at-msa.microscopy.com 4, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 4, 18 -- Subject: FE SEM recommendations 4, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 18 -- X-Spam-Score: 0.00% 4, 18 -- X-MASF: 0.00% 4, 18 -- X-Whitelist: 0.00% ==============================End of - Headers==============================
Thanks for bringing up the topic. I am newly returned to this listserver so missed the earlier discussion. Since I am reluctant to get yet another ID I will post my thoughts here.
I read the Science News article listed below and agree with the conclusions of the authors of the JAMA study. Cheers, Roseann
From Science News, Vol. 169, No. 18, May 6, 2006, p. 285., There is an article titled: Study finds bias in peer review. Abstract: Researchers have found evidence of bias when scientists review data and the researcher's name and affiliation are available to the reviewers.
The synopsis was of: JAMA. 2006;295:1675-1680. The authors of the study concluded:
Our study provides evidence of reviewer bias in the open review of abstracts, favoring authors from the United States, from English-speaking countries outside the United States, and from prestigious academic institutions and likely favoring authors from US government agencies and not from private industry. Also, blinded review at least partially reduces bias.
Our results suggest that adoption of blinded peer review by scientific research meetings is a reasonable, low-cost intervention with substantial benefit.
Roseann Csencsits, PhD TEM Facility Manager Donner Lab Lawrence Berkeley Laboratory 510-486-4548
==============================Original Headers============================== 11, 21 -- From RCsencsits-at-lbl.gov Mon Oct 30 15:12:55 2006 11, 21 -- Received: from mta1.lbl.gov (mta1.lbl.gov [128.3.41.24]) 11, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9ULCta1022662 11, 21 -- for {microscopy-at-microscopy.com} ; Mon, 30 Oct 2006 15:12:55 -0600 11, 21 -- Received: from mta1.lbl.gov (localhost [127.0.0.1]) 11, 21 -- by mta1.lbl.gov (8.13.8/8.13.8) with ESMTP id k9ULCrD7017371 11, 21 -- for {microscopy-at-microscopy.com} ; Mon, 30 Oct 2006 13:12:54 -0800 (PST) 11, 21 -- Received: from [131.243.35.34] (0-17-f2-2d-d1-b7.dhcp.lbl.gov [131.243.35.34]) 11, 21 -- by mta1.lbl.gov (8.13.8/8.13.8) with ESMTP id k9ULCrja017364 11, 21 -- for {microscopy-at-microscopy.com} ; Mon, 30 Oct 2006 13:12:53 -0800 (PST) 11, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 11, 21 -- Content-Transfer-Encoding: 7bit 11, 21 -- Message-Id: {C888EDEC-B9AD-4319-A271-338D6A7EA41D-at-lbl.gov} 11, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 11, 21 -- To: microscopy-at-microscopy.com 11, 21 -- From: Roseann Csencsits {RCsencsits-at-lbl.gov} 11, 21 -- Subject: Scientific integrity discussion 11, 21 -- Date: Mon, 30 Oct 2006 13:12:15 -0800 11, 21 -- X-Mailer: Apple Mail (2.752.3) 11, 21 -- X-Virus-Scanned: ClamAV 0.88.5/2132/Mon Oct 30 11:42:34 2006 on mta1 11, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
Hi Listers I have a user who is embedding tissue in spurs, cutting sections, etching away the plastic, staining with antibodies/fluorophors and cover-slipping. My question is, are there any recommendations for anti-fades that he should use? Thank you David
==============================Original Headers============================== 1, 20 -- From elliott-at-arizona.edu Mon Oct 30 16:54:27 2006 1, 20 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 1, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9UMsRDc006636 1, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 16:54:27 -0600 1, 20 -- Received: from localhost (eowyn.email.arizona.edu [10.0.0.221]) 1, 20 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 1196B11C6E90 1, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 15:54:27 -0700 (MST) 1, 20 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 1, 20 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 4C79511C703B 1, 20 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 15:54:26 -0700 (MST) 1, 20 -- Mime-Version: 1.0 (Apple Message framework v752.2) 1, 20 -- Content-Transfer-Encoding: 7bit 1, 20 -- Message-Id: {FFA25C40-FD8A-4714-B26C-55AC79F17379-at-arizona.edu} 1, 20 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 1, 20 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 1, 20 -- From: David Elliott {elliott-at-arizona.edu} 1, 20 -- Subject: anti-fade for spurs sections 1, 20 -- Date: Mon, 30 Oct 2006 15:54:25 -0700 1, 20 -- X-Mailer: Apple Mail (2.752.2) 1, 20 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
What type of stainless steel are you preparing. If you have a martensitic stainless steel with any substantial hardness, you will have trouble. If you have a austenitic that is annealed, you will have less of a problem. I have a technique for preparing M50 and 440C substrates, but it is rather involved, so let me know. You have to prevent the sample for distorting because of the high internal stresses in the material.
If you don't have the martensitic SS, then there are a number of ways that you can prepare your samples. One would be to make a stack, slice it, core drill the samples, dimple and then ion mill. Another choice would be to use the Tripod Polisher(R) technique followed by ion milling. And a third method would be to use the Technoorg-Linda method for preparing the samples. All of these have been used to prepare samples of coatings on metals.
Disclaimer: South Bay Technology makes and sells much of the equipment that can be used in these techniques and we can help you with any of them and help you select the best one that may be suited for your laboratory. We have a number of technical papers and application notes available on our website for downloading. If you would like to discuss any of these techniques with me, please feel free to email or phone me.
South Bay Technology also represents Technoorg-Linda in the United States.
-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: hikonishi-at-gmail.com [mailto:hikonishi-at-gmail.com] Sent: Monday, October 30, 2006 9:22 AM To: Walck-at-SouthBayTech.com
Dear List members: Oct 30, 2006
I would like to know how to prepare cross section samples of multilayers on stainless steel substrate. I have the basic knowledge on the sample preparation of layers on silicon wafer using M-bond and ion thinning. However, stainless steel will be difficult to cut and grind. My sample is multilayers (such as Al2O3) on stainless steel substrate, and the thickness of multi-layers is around 3 um. Each layer has several 10 to 1 um in thickness. The other side of the multilayers is also stainless steel. The multi-layers are sandwiched by stainless steel. I will have 5 X 5 X 5 mm cubes. I will try to make TEM samples using ion-thining, and later try to use FIB. I would be appreciate if you could provide me any suggestions.
Thank you,
Hiromi Konishi, Ph.D. Laboratory Manager The S.W. Bailey X-ray Diffraction Laboratory Room A353 Weeks Hall Voice: (608) 262-9784, (608) 262-0915 Fax: (608) 262-0693 Department of Geology and Geophysics University of Wisconsin-Madison 1215 W Dayton St. Madison WI 53706
==============================Original Headers============================== 5, 30 -- From hikonishi-at-gmail.com Mon Oct 30 11:12:44 2006 5, 30 -- Received: from nz-out-0102.google.com (nz-out-0102.google.com [64.233.162.196]) 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9UHCiwK029303 5, 30 -- for {Microscopy-at-microscopy.com} ; Mon, 30 Oct 2006 11:12:44 -0600 5, 30 -- Received: by nz-out-0102.google.com with SMTP id r28so1227347nza 5, 30 -- for {Microscopy-at-microscopy.com} ; Mon, 30 Oct 2006 09:12:43 -0800 (PST) 5, 30 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 30 -- s=beta; d=gmail.com; 5, 30 -- h=received:message-id:from:to:subject:date:mime-version:content-type:content -transfer-encoding:x-priority:x-msmail-priority:x-mailer:x-mimeole; 5, 30 -- b=Jldmol8W9oGLiMNFyz0UELzSB1lm5/VlowaOg3Y0nHr0I7KnVF8GbMlMD29ktnvvbjJSre5bWx +j/e+9vY98gl/MYmxBaC4hjFeRf8JAKkyEy39VCZLSop1NN365FHSLItPUzoUYkdL+Mzrpg+wiie bRPrtaQBMN2772y2WoKyE= 5, 30 -- Received: by 10.35.51.19 with SMTP id d19mr5316975pyk; 5, 30 -- Mon, 30 Oct 2006 09:12:43 -0800 (PST) 5, 30 -- Received: from HKONISHI ( [144.92.207.106]) 5, 30 -- by mx.google.com with ESMTP id 19sm12937602nzp.2006.10.30.09.12.41; 5, 30 -- Mon, 30 Oct 2006 09:12:42 -0800 (PST) 5, 30 -- Message-ID: {003201c6fc46$a2504e40$6acf5c90-at-HKONISHI} 5, 30 -- From: "Hiromi Konishi" {hikonishi-at-gmail.com} 5, 30 -- To: {Microscopy-at-microscopy.com} 5, 30 -- Subject: cross section of multilayers on stainless substrate 5, 30 -- Date: Mon, 30 Oct 2006 11:12:47 -0600 5, 30 -- MIME-Version: 1.0 5, 30 -- Content-Type: text/plain; 5, 30 -- format=flowed; 5, 30 -- charset="iso-2022-jp"; 5, 30 -- reply-type=original 5, 30 -- Content-Transfer-Encoding: 7bit 5, 30 -- X-Priority: 3 5, 30 -- X-MSMail-Priority: Normal 5, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 5, 30 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
==============================Original Headers============================== 18, 22 -- From walck-at-southbaytech.com Mon Oct 30 17:12:05 2006 18, 22 -- Received: from ylpvm01.prodigy.net (ylpvm01-ext.prodigy.net [207.115.57.32]) 18, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9UNC4RM017241 18, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 30 Oct 2006 17:12:05 -0600 18, 22 -- X-ORBL: [64.169.193.90] 18, 22 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 18, 22 -- by ylpvm01.prodigy.net (8.13.7 out spool5000 dk/8.13.7) with ESMTP id k9UNAvB5012008; 18, 22 -- Mon, 30 Oct 2006 18:10:58 -0500 18, 22 -- From: "Scott Walck" {walck-at-southbaytech.com} 18, 22 -- To: {Microscopy-at-microscopy.com} 18, 22 -- Cc: {hikonishi-at-gmail.com} 18, 22 -- Subject: RE: [Microscopy] cross section of multilayers on stainless substrate 18, 22 -- Date: Mon, 30 Oct 2006 15:12:30 -0800 18, 22 -- Message-ID: {015b01c6fc78$e003ac50$7801a8c0-at-dynamicbl8uno3} 18, 22 -- MIME-Version: 1.0 18, 22 -- Content-Type: text/plain; 18, 22 -- charset="us-ascii" 18, 22 -- Content-Transfer-Encoding: 7bit 18, 22 -- X-Mailer: Microsoft Office Outlook 11 18, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 18, 22 -- Thread-Index: Acb8R/PPjHCGn3xCSgueB0JhtIHrGwAKDPuw 18, 22 -- In-reply-to: {200610301722.k9UHMIp6002965-at-ns.microscopy.com} ==============================End of - Headers==============================
That may be true, but the person sees quite nice, specific, multi- channel staining (controls in place). I was surprised.
That said, any recommendations for an anti-fade?
David
On Oct 30, 2006, at 4:38 PM, heckman-at-bgnet.bgsu.edu wrote:
} David- } It would be amazing if the person saw any specific staining. Spurr's } is almost impenetrable when it comes to putting a big molecule like an } antibody into the polymerized material. } Carol } }
==============================Original Headers============================== 7, 22 -- From elliott-at-arizona.edu Mon Oct 30 21:17:05 2006 7, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9V3H5Z3000446 7, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 21:17:05 -0600 7, 22 -- Received: from localhost (faramir.email.arizona.edu [10.0.0.218]) 7, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id D2E6211C7F81 7, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 20:17:04 -0700 (MST) 7, 22 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 7, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id C1F6711C81AD 7, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 20:17:03 -0700 (MST) 7, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 7, 22 -- In-Reply-To: {1162251520-26358.041.123-smmsdV2.1.2-at-smtp.bgsu.edu} 7, 22 -- References: {1162251520-26358.041.123-smmsdV2.1.2-at-smtp.bgsu.edu} 7, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 7, 22 -- Message-Id: {59D6BD8D-00AE-4957-A48E-D036C9CD94D5-at-arizona.edu} 7, 22 -- Content-Transfer-Encoding: 7bit 7, 22 -- From: David Elliott {elliott-at-arizona.edu} 7, 22 -- Subject: Re: [Microscopy] anti-fade for spurs sections 7, 22 -- Date: Mon, 30 Oct 2006 20:17:01 -0700 7, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 7, 22 -- X-Mailer: Apple Mail (2.752.2) 7, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
David, Maybe there is a reason for using Spurrs, but if your user would like an alternative that will certainly make the etching etching, he or she might like to investigate the butyl methyl methacrylate system that I have worked on for years. Its easy and works well. If your user is interested, I can send some references and a protocol. Contact me off-line.
Tobias
} } Hi Listers } I have a user who is embedding tissue in spurs, cutting sections, } etching away the plastic, staining with antibodies/fluorophors and } cover-slipping. My question is, are there any recommendations for } anti-fades that he should use? } Thank you } David }
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Email: dmcdaniel-at-usuhs.mil Name: Dennis McDaniel
Title-Subject: [Filtered] LR White flat embedding
Question: I have some fibroblasts which were grown on glass coverslips and fixed, dehydrated and infiltrated with LR White. I put a piece of aclar film over the top and polymerized the resin, but now I am having a very difficult time getting the LR White to separate from the coverslip so that I can glue it to an empty BEEM capsule. Does anyone know any good tricks? Thanks.
Two thoughts; 1) not helpful now, but have the cells grown on aclar rather than glass in the future 2) try dry-ice on the glass. This often helps remove the glass, sometimes it just shatters the glass leaving the plastic behind. I have had mixed results. David
On Oct 31, 2006, at 7:59 AM, dmcdaniel-at-usuhs.mil 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 dmcdaniel-at-usuhs.mil as well as the MIcroscopy } Listserver } ---------------------------------------------------------------------- } ----- } } Email: dmcdaniel-at-usuhs.mil } Name: Dennis McDaniel } } Title-Subject: [Filtered] LR White flat embedding } } Question: I have some fibroblasts which were grown on glass } coverslips and fixed, dehydrated and infiltrated with LR White. I } put a piece of aclar film over the top and polymerized the resin, } but now I am having a very difficult time getting the LR White to } separate from the coverslip so that I can glue it to an empty BEEM } capsule. Does anyone know any good tricks? Thanks. } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== } 5, 12 -- From zaluzec-at-microscopy.com Tue Oct 31 08:55:01 2006 } 5, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com } [206.69.208.22]) } 5, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP } id k9VEt18Q002472 } 5, 12 -- for {microscopy-at-microscopy.com} ; Tue, 31 Oct 2006 } 08:55:01 -0600 } 5, 12 -- Mime-Version: 1.0 } 5, 12 -- X-Sender: (Unverified) } 5, 12 -- Message-Id: {p06110409c16d14313ef7-at-[206.69.208.22]} } 5, 12 -- Date: Tue, 31 Oct 2006 08:54:59 -0600 } 5, 12 -- To: microscopy-at-microscopy.com } 5, 12 -- From: dmcdaniel-at-usuhs.mil (by way of MicroscopyListserver) } 5, 12 -- Subject: viaWWW: LR White flat embedding } 5, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers==============================
==============================Original Headers============================== 5, 22 -- From Elliott-at-arizona.edu Tue Oct 31 09:07:06 2006 5, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9VF76wv013078 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 31 Oct 2006 09:07:06 -0600 5, 22 -- Received: from localhost (gimli.email.arizona.edu [10.0.0.223]) 5, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id DF4E911C93AD 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 31 Oct 2006 08:07:03 -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 2103C11C9482 5, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 31 Oct 2006 08:06:42 -0700 (MST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 22 -- In-Reply-To: {200610311459.k9VEx1fP007748-at-ns.microscopy.com} 5, 22 -- References: {200610311459.k9VEx1fP007748-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 22 -- Message-Id: {FF4005FF-CD59-4641-8C3D-EFA32B704C07-at-arizona.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: David Elliott {Elliott-at-arizona.edu} 5, 22 -- Subject: Re: [Microscopy] viaWWW: LR White flat embedding 5, 22 -- Date: Tue, 31 Oct 2006 08:06:40 -0700 5, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 5, 22 -- X-Mailer: Apple Mail (2.752.2) 5, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
When separating glass from resin I use liquid nitrogen. I dip the coverslip about halfway and hold there for just a few seconds. Then I hold the slip and resin combination between my finger and thumb to begin warming. It seems the warming happens at a different rate and causes the layers to separate. You can then slip the glass away from the resin. Sometimes the glass shatters, but you can still slip it off in pieces.
In future, try growing your fibroblasts on Thermanox coverslips. It works very well and the cells grow just fine. Invert the coverslip over a drop of resin on ACLAR and polymerize. The coverslip and ACLAR are removed easily from the resin. Even if the resin sticks slightly to the coverslip the layer is so thin that you can simply use a razor blade to cut and remove the precise area of the cells that you would like to look at. The rest of the cells can be saved for another time.
Good luck, Jo Dee
Jo Dee Fish Research Technologist III Gladstone Institute of Cardiovascular Disease
Mailing address: The J. David Gladstone Institutes 1650 Owens Street San Francisco, CA 94158
-----Original Message----- X-from: dmcdaniel-at-usuhs.mil [mailto:dmcdaniel-at-usuhs.mil] Sent: Tuesday, October 31, 2006 6:59 AM To: jfish-at-gladstone.ucsf.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 dmcdaniel-at-usuhs.mil as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: dmcdaniel-at-usuhs.mil Name: Dennis McDaniel
Title-Subject: [Filtered] LR White flat embedding
Question: I have some fibroblasts which were grown on glass coverslips and fixed, dehydrated and infiltrated with LR White. I put a piece of aclar film over the top and polymerized the resin, but now I am having a very difficult time getting the LR White to separate from the coverslip so that I can glue it to an empty BEEM capsule. Does anyone know any good tricks? Thanks.
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Don't know why you think stainless steel should be difficult to cut and grind - it's a lot easier than silicon!
Have you considered spark errosion methods? Most (all?) steels can be cut readily by spark errosion. I would suggest that you follow a similar route to the one you are familiar with for preparing cross-sections from layers on Si wafer except:
1. When cutting the cylindrical core, use a similar trepanning tool but in a spark errosion system.
2. Having glued the core in to the Cu cylinder, slice by spark errosion with moving wire.
You then have 3 mm discs which are easily thinned further by mechanical grinding, if necessary, followed by dimple grinding and polishing.
Final perforation can be achieved either by ion beam thinning or electropolishing - if you layers don't conduct, ion beam thinning is probably better.
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==============================Original Headers============================== 9, 17 -- From larry-at-cymru.freewire.co.uk Tue Oct 31 15:13:10 2006 9, 17 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9VLD9nD010322 9, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 31 Oct 2006 15:13:10 -0600 9, 17 -- Received: from [217.154.250.226] (th6dc-217-154-250-226.dial.mistral.co.uk [217.154.250.226] (may be forged)) 9, 17 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id k9VLD6j9003897; 9, 17 -- Tue, 31 Oct 2006 21:13:08 GMT 9, 17 -- Mime-Version: 1.0 9, 17 -- Message-Id: {p06210200c16d6605f1ac-at-[217.154.250.72]} 9, 17 -- In-Reply-To: {200610301716.k9UHGJm4031554-at-ns.microscopy.com} 9, 17 -- References: {200610301716.k9UHGJm4031554-at-ns.microscopy.com} 9, 17 -- Date: Tue, 31 Oct 2006 20:57:31 +0000 9, 17 -- To: hikonishi-at-gmail.com, Microscopy-at-MSA.Microscopy.Com 9, 17 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 9, 17 -- Subject: Re: [Microscopy] cross section of multilayers on stainless 9, 17 -- substrate 9, 17 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
David, I suggest 2% n-propyl gallate in 100% glycerol. I have used it as an anti-fade with many types of samples. It works as well as any commercial product that I have tried. It is easy to make, just stir overnight. It is inexpensive. I use fingernail polish to affix a coverslip to the slide. see the following reference Giloh H, Sedat JW. Fluorescence microscopy: reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propyl gallate. Science. 1982 Sep 24;217(4566):1252-5. Larry
elliott-at-arizona.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 } ---------------------------------------------------------------------------- } } That may be true, but the person sees quite nice, specific, multi- } channel staining (controls in place). I was surprised. } } That said, any recommendations for an anti-fade? } } David } } } On Oct 30, 2006, at 4:38 PM, heckman-at-bgnet.bgsu.edu wrote: } } } } David- } } It would be amazing if the person saw any specific staining. Spurr's } } is almost impenetrable when it comes to putting a big molecule like an } } antibody into the polymerized material. } } Carol } } } } } } } } ==============================Original Headers============================== } 7, 22 -- From elliott-at-arizona.edu Mon Oct 30 21:17:05 2006 } 7, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) } 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9V3H5Z3000446 } 7, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 21:17:05 -0600 } 7, 22 -- Received: from localhost (faramir.email.arizona.edu [10.0.0.218]) } 7, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id D2E6211C7F81 } 7, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 20:17:04 -0700 (MST) } 7, 22 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) } 7, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id C1F6711C81AD } 7, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 30 Oct 2006 20:17:03 -0700 (MST) } 7, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) } 7, 22 -- In-Reply-To: {1162251520-26358.041.123-smmsdV2.1.2-at-smtp.bgsu.edu} } 7, 22 -- References: {1162251520-26358.041.123-smmsdV2.1.2-at-smtp.bgsu.edu} } 7, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 7, 22 -- Message-Id: {59D6BD8D-00AE-4957-A48E-D036C9CD94D5-at-arizona.edu} } 7, 22 -- Content-Transfer-Encoding: 7bit } 7, 22 -- From: David Elliott {elliott-at-arizona.edu} } 7, 22 -- Subject: Re: [Microscopy] anti-fade for spurs sections } 7, 22 -- Date: Mon, 30 Oct 2006 20:17:01 -0700 } 7, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} } 7, 22 -- X-Mailer: Apple Mail (2.752.2) } 7, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu } ==============================End of - Headers============================== }
-- Larry Ackerman, Associate Specialist UCSF, Dept. of Anatomy, Rm S1347 513 Parnassus Ave., Box 0452 San Francisco, CA 94143
larry.ackerman-at-ucsf.edu
==============================Original Headers============================== 5, 30 -- From Larry.Ackerman-at-ucsf.edu Tue Oct 31 16:37:22 2006 5, 30 -- Received: from emfmcb01.ucsfmedicalcenter.org (emfmcb01.ucsfmedicalcenter.org [64.54.46.97]) 5, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k9VMbMQR022501 5, 30 -- for {Microscopy-at-microscopy.com} ; Tue, 31 Oct 2006 16:37:22 -0600 5, 30 -- Received: from 64.54.128.151 by emfmcb02.ucsfmedicalcenter.org with 5, 30 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 5, 30 -- Tue, 31 Oct 2006 14:49:16 -0800 5, 30 -- X-Server-Uuid: 44B63953-633F-4500-B2DA-A3E478718104 5, 30 -- Received: from [128.218.123.88] ([128.218.123.88]) by 5, 30 -- exvs06.net.ucsf.edu with Microsoft SMTPSVC(6.0.3790.1830); Tue, 31 Oct 5, 30 -- 2006 14:37:13 -0800 5, 30 -- Message-ID: {4547D008.8090700-at-ucsf.edu} 5, 30 -- Date: Tue, 31 Oct 2006 14:36:56 -0800 5, 30 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 5, 30 -- Reply-to: larry.ackerman-at-ucsf.edu 5, 30 -- Organization: UCSF, NeuroAnatomy 5, 30 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 5, 30 -- X-Accept-Language: en-us, en 5, 30 -- MIME-Version: 1.0 5, 30 -- To: Microscopy-at-microscopy.com 5, 30 -- Subject: Re: [Microscopy] Re: anti-fade for spurs sections 5, 30 -- References: {200610310322.k9V3MAij008163-at-ns.microscopy.com} 5, 30 -- In-Reply-To: {200610310322.k9V3MAij008163-at-ns.microscopy.com} 5, 30 -- X-OriginalArrivalTime: 31 Oct 2006 22:37:13.0624 (UTC) 5, 30 -- FILETIME=[1C4C6980:01C6FD3D] 5, 30 -- X-WSS-ID: 69590D661IO2901398-01-01 5, 30 -- Content-Type: text/plain; 5, 30 -- charset=iso-8859-1; 5, 30 -- format=flowed 5, 30 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Here is the November 2006 Microscopy Today table of contents. I will close the subscription list for this issue on Monday, Nov 6th, 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 in Art Galleries? Stephen W. Carmichael, Mayo Clinic
Recent Advances in High-Speed Orientation Mapping Matthew M. Nowell1, Martina Chui-Sabourin2, and John O. Carpenter1 1 EDAX-TSL, Draper, UT, 2 EDAX-TSL, Mahwah, NJ
Diffracted Light Contrast: Improving the Resolution of a Basic Light Microscope by an Order of Magnitude W. Barry Piekos, Yale University, New Haven, CT
Nature’s Engineering Marvels: the Structure and Chemistry of a Butterfly Wing V.S. Smentkowski, S.G. Ostrowski, E.J. Olson, J. Cournoyer, K. Dovidenko, R.A. Potyrailo, General Electric Niskayuna, NY
Integrating High Resolution Light Microscopy and Real Time Observation of Fluorescent Labels Thomas A. Hasling, Aetos Technologies, Inc., Auburn, AL
Surface Rippling & Ion Etch Yields of Diamond Using a Focused Ion Beam: With or Without Enhanced-Chemistry, Aspect Ratio Regulates Ion Etching W. J. MoberlyChan, T. E. Felter, & M. A. Wall, Lawrence Livermore National Lab., Livermore, CA
Internet-Based Administration of Shared Instruments with Facility Online Manager Shu-You Li and Vinayak P. Dravid, Northwestern University, Evanston, IL
Andrew Paul Leonard: Capturing the Cover of Time Magazine Lise Millay Stevens, MA
Streamlining the Modern Lab Radhika Subramanian, Cornet Technology, Inc., Springfield, VA
Intermediate Magnification Imaging System for Whole Organs/Organisms Richard W. Cole, Carmen A. Mannella, Christian Renken, and James N. Turner, Wadsworth Center, Albany, NY
The NEST Laboratory: The Art of a Multi-User Facility Scott Streiker and Rachel Smith, University of Dayton, Dayton, OH
Butyl-methyl-methacrylate for Immunocytochemistry Through the Light Microscope Tobias I. Baskin, University of Massachusetts, Amherst, MA
How To Stick Loosely Adherent Cells To Glass Slides Martin Spitaler, Imperial College, London, UK
Just Say NO to Microtoming Silicon! Ron Anderson, Microscopy Today, Largo, FL
Industry News
Netnotes SPECIMEN PREPARATION - ventricle embedding problem SPECIMEN PREPARATION - sample preparation polymer blend SPECIMEN PREPARATION - Staining starch in sections SPECIMEN PREPARATION - staining methods SPECIMEN PREPARATION - Thiocarbohydrazide SPECIMEN PREPARATION - cotton fibers IMMUNOCYTOCHEMISTRY - Immunogold labeling & SEM IMMUNOCYTOCHEMISTRY – testing colloidal gold IMAGE ANALYSIS – object size EM - Venting EM Chambers TEM – electron diffraction TEM – Nickel grids & EDX SEM – Back scattered electrons and edge effects SEM – Backscattered electron images
Index of Advertisers
==============================Original Headers============================== 21, 18 -- From microscopytoday-at-tampabay.rr.com Wed Nov 1 12:34:25 2006 21, 18 -- Received: from ms-smtp-02.tampabay.rr.com (ms-smtp-02.tampabay.rr.com [65.32.5.132]) 21, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA1IYO6A024688 21, 18 -- for {Microscopy-at-Microscopy.Com} ; Wed, 1 Nov 2006 12:34:25 -0600 21, 18 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 21, 18 -- by ms-smtp-02.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id kA1IYK4Y002917; 21, 18 -- Wed, 1 Nov 2006 13:34:22 -0500 (EST) 21, 18 -- Message-ID: {4548E8AB.4000108-at-tampabay.rr.com} 21, 18 -- Date: Wed, 01 Nov 2006 13:34:19 -0500 21, 18 -- From: Microscopy Today {microscopytoday-at-tampabay.rr.com} 21, 18 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 21, 18 -- MIME-Version: 1.0 21, 18 -- To: Listserver {Microscopy-at-Microscopy.Com} , 21, 18 -- Confocal Listserver {confocal-at-listserv.buffalo.edu} 21, 18 -- Subject: Microscopy Today November 2006 Table of Contents 21, 18 -- Content-Type: text/plain; charset=windows-1252; format=flowed 21, 18 -- Content-Transfer-Encoding: 8bit 21, 18 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
I have a 1972 Edwards 306 coater, with the 'SEM Planetary Workholder', which is used to carbon-coat samples for EPMA analysis.
It often works OK, and I monitor the coating thickness by watching the color change of a polished brass surface which is in there with the samples. Sometimes to get a satisfactory coating I have to go through two or three cycles of: new carbon rod, evacuate, apply current to the rod until it burns out, bring up to atmospheric pressure, allow the rodholder to cool, which drives me a bit nuts, but it always gets there in the end.
It will coat up to three 47 x 27mm slides or six 1" diameter plugs, or 3 of each, at the same time.
I have two questions:
1 On the very left-hand side of the circuit diagram, the auxiliary contacts of 8-Amp circuit breaker CB2 supply mains power, through a few interlock switches, to a rectangular box labelled 'ILC'. What is this ILC thing? I have pored over the manual, searching for a clue, but have found none. Does anyone know either what ILC is, where it is located, or what current it draws? My problem is that CB2 has started to trip out from time to time, I would like to replace CB2, but I have no idea of the current rating needed for the auxiliary contacts. Also, what is the rectangular box 'AAV' just to the left of the Pirani gauge?
2 I have a window of opportunity to replace the whole unit, but the window will be open for only a short while, and I am not familiar with the current market in vacuum coaters. I would greatly appreciate any specific recommendations (directly to me, if you wish) for suitable replacements. The window is not open very wide, either, so my budget is limited to around US$35K.
thanks in anticipation
Ritchie Sims
-- 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 Wed Nov 1 13:22:49 2006 16, 26 -- Received: from groucho.itss.auckland.ac.nz (groucho.itss.auckland.ac.nz [130.216.190.11]) 16, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA1JMlAC004287 16, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 1 Nov 2006 13:22:48 -0600 16, 26 -- Received: from localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) 16, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 7FB8137933 16, 26 -- for {microscopy-at-msa.microscopy.com} ; Thu, 2 Nov 2006 08:22:45 +1300 (NZDT) 16, 26 -- Received: from groucho.itss.auckland.ac.nz ([127.0.0.1]) 16, 26 -- by localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 16, 26 -- with ESMTP id 04177-19 for {microscopy-at-msa.microscopy.com} ; 16, 26 -- Thu, 2 Nov 2006 08:22:45 +1300 (NZDT) 16, 26 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 16, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 80F1937AE3 16, 26 -- for {microscopy-at-msa.microscopy.com} ; Thu, 2 Nov 2006 08:21:07 +1300 (NZDT) 16, 26 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 16, 26 -- To: microscopy-at-msa.microscopy.com 16, 26 -- Date: Thu, 02 Nov 2006 08:21:06 +1300 16, 26 -- MIME-Version: 1.0 16, 26 -- Subject: Carbon Coater questions 16, 26 -- Message-ID: {4549AA72.13398.2AA396-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==============================
The problem is potentially due to the carbon rods rather than the evaporator. Brown et al. 1991 (Carbon rod failure during carbon coating, The Microscope, v. 39, pp265-267) describe how the quality/composition of the carbon rod can significantly effect the longevity of carbon rods during carbon evaporation.
Cheers, Bryan Bandli
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 } ---------------------------------------------------------------------------- } } } Hi } } I have a 1972 Edwards 306 coater, with the 'SEM Planetary } Workholder', which is used to carbon-coat samples for EPMA analysis. } } It often works OK, and I monitor the coating thickness by watching the } color change of a polished brass surface which is in there with the } samples. Sometimes to get a satisfactory coating I have to go through } two or three cycles of: new carbon rod, evacuate, apply current to the } rod until it burns out, bring up to atmospheric pressure, allow the } rodholder to cool, which drives me a bit nuts, but it always gets there in } the end. } } It will coat up to three 47 x 27mm slides or six 1" diameter plugs, or 3 } of each, at the same time. } } I have two questions: } } 1 On the very left-hand side of the circuit diagram, the auxiliary } contacts of 8-Amp circuit breaker CB2 supply mains power, through a } few interlock switches, to a rectangular box labelled 'ILC'. What is this } ILC thing? I have pored over the manual, searching for a clue, but have } found none. Does anyone know either what ILC is, where it is located, } or what current it draws? My problem is that CB2 has started to trip out } from time to time, I would like to replace CB2, but I have no idea of the } current rating needed for the auxiliary contacts. Also, what is the } rectangular box 'AAV' just to the left of the Pirani gauge? } } 2 I have a window of opportunity to replace the whole unit, but the } window will be open for only a short while, and I am not familiar with } the current market in vacuum coaters. I would greatly appreciate any } specific recommendations (directly to me, if you wish) for suitable } replacements. The window is not open very wide, either, so my budget } is limited to around US$35K. } } } thanks in anticipation } } } Ritchie Sims } } } } -- } 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 Wed Nov 1 13:22:49 2006 } 16, 26 -- Received: from groucho.itss.auckland.ac.nz (groucho.itss.auckland.ac.nz [130.216.190.11]) } 16, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA1JMlAC004287 } 16, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 1 Nov 2006 13:22:48 -0600 } 16, 26 -- Received: from localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) } 16, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 7FB8137933 } 16, 26 -- for {microscopy-at-msa.microscopy.com} ; Thu, 2 Nov 2006 08:22:45 +1300 (NZDT) } 16, 26 -- Received: from groucho.itss.auckland.ac.nz ([127.0.0.1]) } 16, 26 -- by localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) } 16, 26 -- with ESMTP id 04177-19 for {microscopy-at-msa.microscopy.com} ; } 16, 26 -- Thu, 2 Nov 2006 08:22:45 +1300 (NZDT) } 16, 26 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) } 16, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 80F1937AE3 } 16, 26 -- for {microscopy-at-msa.microscopy.com} ; Thu, 2 Nov 2006 08:21:07 +1300 (NZDT) } 16, 26 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} } 16, 26 -- To: microscopy-at-msa.microscopy.com } 16, 26 -- Date: Thu, 02 Nov 2006 08:21:06 +1300 } 16, 26 -- MIME-Version: 1.0 } 16, 26 -- Subject: Carbon Coater questions } 16, 26 -- Message-ID: {4549AA72.13398.2AA396-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============================== } }
-- ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- Bryan Bandli Research Microscopist MVA Scientific Consultants 3300 Breckinridge Blvd., Suite 400 (770) 662-8509 ----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------- The information in this email is confidential and may be legally privileged. It is intended solely for the addressee. If you are not the intended recipient, please delete the email and notify MVA Scientific Consultants of the transmission error. MVA Scientific Consultants - 3300 Breckinridge Blvd. Suite 400, Duluth, GA 30096 - (770)662-8509 -------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------
==============================Original Headers============================== 7, 20 -- From bbandli-at-mvainc.com Wed Nov 1 13:42:36 2006 7, 20 -- Received: from smtp02.atlngahp.sys.nuvox.net (smtp-out2.atlngahp.sys.nuvox.net [70.43.63.19]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA1JgYjP015107 7, 20 -- for {microscopy-at-msa.microscopy.com} ; Wed, 1 Nov 2006 13:42:35 -0600 7, 20 -- Received: from [192.168.1.95] (216.215.228.34.nw.nuvox.net [216.215.228.34]) 7, 20 -- by smtp02.atlngahp.sys.nuvox.net (8.13.1/8.13.1) with ESMTP id kA1JgOKH016050; 7, 20 -- Wed, 1 Nov 2006 14:42:25 -0500 7, 20 -- Message-ID: {4548F8A6.8040203-at-mvainc.com} 7, 20 -- Date: Wed, 01 Nov 2006 14:42:30 -0500 7, 20 -- From: bbandli {bbandli-at-mvainc.com} 7, 20 -- Reply-To: bbandli-at-mvainc.com 7, 20 -- Organization: MVA Scientific Consultants 7, 20 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 7, 20 -- MIME-Version: 1.0 7, 20 -- To: r.sims-at-auckland.ac.nz, microscopy-at-msa.microscopy.com 7, 20 -- Subject: Re: [Microscopy] Carbon Coater questions 7, 20 -- References: {200611011923.kA1JNMMa005391-at-ns.microscopy.com} 7, 20 -- In-Reply-To: {200611011923.kA1JNMMa005391-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==============================
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Title-Subject: [Filtered] TEM- EDS on vitrified samples
Question: I'm trying to determine whether EDS analysis can be performed successfuly on vitrified cryo TEM samples. I have asked around, and done a few brief literature searches for information on this topic, and haven't got any good answers.
If anyone has any experience in this area, I'd love to hear from you.
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Email: dmcdaniel-at-usuhs.mil Name: Dennis McDaniel
Title-Subject: [Filtered] LR White flat embedding
Question: I have some fibroblasts which were grown on glass coverslips and fixed, dehydrated and infiltrated with LR White. I put a piece of aclar film over the top and polymerized the resin, but now I am having a very difficult time getting the LR White to separate from the coverslip so that I can glue it to an empty BEEM capsule. Does anyone know any good tricks? Thanks.
I have a box here with EM400 analytical pole pieces for +/- 60 degree tilt. At least that's what it says on the box. Is anyone interested in having them? Kim
-- Kim Rensing Ph.D. Manager, Microscopy and Imaging Facility
University of Calgary, Health Sciences Centre B129 - 3330 Hospital Drive NW Calgary, AB, Canada T2N 4N1
403-220-3488 krensing-at-ucalgary.ca
==============================Original Headers============================== 5, 25 -- From krensing-at-ucalgary.ca Thu Nov 2 10:52:48 2006 5, 25 -- Received: from mr1.ucalgary.ca (mr1.ucalgary.ca [136.159.34.165]) 5, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA2Gqj3c007053 5, 25 -- for {Microscopy-at-Microscopy.Com} ; Thu, 2 Nov 2006 10:52:48 -0600 5, 25 -- Received: from smtp2.ucalgary.ca (smtp2.ucalgary.ca [136.159.34.223]) 5, 25 -- by mr1.ucalgary.ca (Postfix) with ESMTP id 8A3157CA8 5, 25 -- for {Microscopy-at-Microscopy.Com} ; Thu, 2 Nov 2006 09:52:44 -0700 (MST) 5, 25 -- Received: from [192.168.0.100] ([136.159.164.171]) 5, 25 -- (authenticated (0 bits)) 5, 25 -- by smtp2.ucalgary.ca (8.11.7/8.11.6) with ESMTP id kA2GqhQ11327 5, 25 -- (using TLSv1/SSLv3 with cipher DHE-RSA-AES256-SHA (256 bits) verified NO) 5, 25 -- for {Microscopy-at-Microscopy.Com} ; Thu, 2 Nov 2006 09:52:43 -0700 5, 25 -- Message-ID: {454A2259.4060909-at-ucalgary.ca} 5, 25 -- Date: Thu, 02 Nov 2006 09:52:41 -0700 5, 25 -- From: Kim Rensing {krensing-at-ucalgary.ca} 5, 25 -- Organization: Microscopy and Imaging Facility, U. of Calgary 5, 25 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 5, 25 -- MIME-Version: 1.0 5, 25 -- To: Microscopy-at-Microscopy.Com 5, 25 -- Subject: EM400 pole pieces 5, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 5, 25 -- Content-Transfer-Encoding: 7bit 5, 25 -- X-UCalgary-MailScanner-Information: Please contact IT Help Desk at (403) 220-5555 for more information 5, 25 -- X-UCalgary-MailScanner: Found to be clean 5, 25 -- X-UCalgary-MailScanner-From: krensing-at-ucalgary.ca ==============================End of - Headers==============================
My samples are indeed particles, which are in an aqueous suspension. The suspensions are vitrified for cryo TEM imaging to observe particles in the pseudo-liquid state. The goal with EDS would be to get elemental mapping of the particles themselves. What I am concerned about is the degradation of the sample upon analysis with EDS, and if enough signal can be detected from particles that may be only 100 nm in diameter.
Our ultimate goal is to characterize differences between the surface of the particle and it's interior, in the solution state. To my limited knowledge, there is not a characterization technique out there that specifically addresses this problem. If there is, I'd be interested in hearing of it.
Thank you, Jessica
-----Original Message----- X-from: Chaoying Ni [mailto:cni-at-UDel.Edu] Sent: Thursday, November 02, 2006 6:12 AM To: Cervantes, Jessica
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Title-Subject: [Filtered] TEM- EDS on vitrified samples
Question: I'm trying to determine whether EDS analysis can be performed successfuly on vitrified cryo TEM samples. I have asked around, and done a few brief literature searches for information on this topic, and haven't got any good answers.
If anyone has any experience in this area, I'd love to hear from you.
Thanks, Jessica Cervantes
==============================Original Headers============================== 21, 14 -- From cervantes-at-bendres.com Thu Nov 2 11:57:34 2006 21, 14 -- Received: from BRIEX04A.bri.local (bc161112.bendcable.com [216.228.161.112]) 21, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA2HvYvp018608 21, 14 -- for {microscopy-at-microscopy.com} ; Thu, 2 Nov 2006 11:57:34 -0600 21, 14 -- MIME-Version: 1.0 21, 14 -- Content-Type: text/plain; 21, 14 -- charset="iso-8859-1" 21, 14 -- Subject: RE: [Microscopy]: TEM- EDS on vitrified samples 21, 14 -- Date: Thu, 2 Nov 2006 09:57:33 -0800 21, 14 -- Message-ID: {8943D65F9AD70E4488AD6DE09F15088768C17F-at-BRIEX04A} 21, 14 -- From: "Cervantes, Jessica" {cervantes-at-bendres.com} 21, 14 -- To: "Chaoying Ni" {cni-at-UDel.Edu} , {microscopy-at-microscopy.com} 21, 14 -- Content-Transfer-Encoding: 8bit 21, 14 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA2HvYvp018608 ==============================End of - Headers==============================
OK. You are right about the possible complication you may run into while you focus a beam onto a particle you want to analyze. I can see the scenario that could happen assuming your particle is non-volatile: amorphous H2O --} crystal H2O + evaporation, left with your particle. I don't see a problem until you have to realize analyzing/mapping the interior and exterior composition of a 100nm solid particle itself sometimes could be a little tricky. If the particle itself is volatile, then good luck and the work should be publishable. -cni
-----Original Message----- X-from: Cervantes, Jessica [mailto:cervantes-at-bendres.com] Sent: Thursday, November 02, 2006 12:58 PM To: Chaoying Ni; microscopy-at-microscopy.com
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Title-Subject: [Filtered] TEM- EDS on vitrified samples
Question: I'm trying to determine whether EDS analysis can be performed successfuly on vitrified cryo TEM samples. I have asked around, and done a few brief literature searches for information on this topic, and haven't got any good answers.
If anyone has any experience in this area, I'd love to hear from you.
Thanks, Jessica Cervantes
==============================Original Headers============================== 25, 26 -- From cni-at-udel.edu Thu Nov 2 12:21:32 2006 25, 26 -- Received: from md3.nss.udel.edu (md3.nss.udel.edu [128.175.1.13]) 25, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA2ILVSE029456 25, 26 -- for {microscopy-at-microscopy.com} ; Thu, 2 Nov 2006 12:21:31 -0600 25, 26 -- Received: from Suba (em076-119.engg.udel.edu [128.175.119.76]) 25, 26 -- by md3.nss.udel.edu (MOS 3.8.2-GA) 25, 26 -- with ESMTP id DLQ55625; 25, 26 -- Thu, 2 Nov 2006 13:21:29 -0500 (EST) 25, 26 -- From: "Chaoying Ni" {cni-at-udel.edu} 25, 26 -- To: "'Cervantes, Jessica'" {cervantes-at-bendres.com} , 25, 26 -- {microscopy-at-microscopy.com} 25, 26 -- Subject: RE: [Microscopy]: TEM- EDS on vitrified samples 25, 26 -- Date: Thu, 2 Nov 2006 13:21:42 -0500 25, 26 -- Organization: University of Delaware 25, 26 -- Message-ID: {000a01c6feab$bf5b6b20$4c77af80-at-Suba} 25, 26 -- MIME-Version: 1.0 25, 26 -- Content-Type: text/plain; 25, 26 -- charset="us-ascii" 25, 26 -- X-Priority: 3 (Normal) 25, 26 -- X-MSMail-Priority: Normal 25, 26 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 25, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 25, 26 -- Importance: Normal 25, 26 -- In-Reply-To: {8943D65F9AD70E4488AD6DE09F15088768C17F-at-BRIEX04A} 25, 26 -- Content-Transfer-Encoding: 8bit 25, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA2ILVSE029456 ==============================End of - Headers==============================
On Nov 1, 2006, at 7:33 PM, cervantes-at-bendres.com wrote:
} Question: I'm trying to determine whether EDS analysis can be } performed successfuly on vitrified cryo TEM samples. I have asked } around, and done a few brief literature searches for information on } this topic, and haven't got any good answers. } } If anyone has any experience in this area, I'd love to hear from you. } Dear Jessica, I did some EDS on the HVEM at Albany on cryo samples, but our set-up was somewhat unusual, so you may have some problems related to the geometry of your instrument. We had our detector mounted so that it looked upward at the specimen at an angle corresponding to a minimum in the brehmsstrahlung distribution, and we tilted the specimen to an angle so that the specimen plane bisected the angle between the beam direction and the detector angle. Our anticontaminator was a blade located between the specimen and the objective aperture--in the HVEM there is a lot of room between the pole pieces. The specimen was a foram with a lot of Si in it, so I got a very good peak and low background. There was essentially no difference between EDS on RT samples and this sample; however, since there is usually a lot of ice and relatively little specimen in a given volume, you may have some S/N problems unless you are looking for an element that is very abundant. 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 Thu Nov 2 13:13:31 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 kA2JDUBI008547 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 2 Nov 2006 13:13:31 -0600 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 534702EF1D 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 2 Nov 2006 11:13:30 -0800 (PST) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id 7B3A235490 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 2 Nov 2006 11:13:24 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200611020333.kA23XvIL002787-at-ns.microscopy.com} 4, 22 -- References: {200611020333.kA23XvIL002787-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {b9f598e6c4674c961550c770691cf76b-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: TEM- EDS on vitrified samples 4, 22 -- Date: Thu, 2 Nov 2006 11:18:23 -0800 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==============================
Jessica, not specifically cryo-TEM, but we performed EDS on lyophilized, vitrified (slam freezing) cryo-sections back in '84. Here is the reference:
Harding CV, Susan S, Bobrowski W Elemental profiles in cryosections and frozen-dried bulk specimens of the normal lens. Ophthalmic Res. 1984;16(5):276-83.
If you need it, I can send you a PDF of the publication. Best regards,
Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"The ultimate human freedom is the ability to choose one's attitude in a given set of circumstances." -Viktor Frankl
Title-Subject: [Filtered] TEM- EDS on vitrified samples
Question: I'm trying to determine whether EDS analysis can be performed successfuly on vitrified cryo TEM samples. I have asked around, and done a few brief literature searches for information on this topic, and haven't got any good answers.
If anyone has any experience in this area, I'd love to hear from you.
Thanks, Jessica Cervantes ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately.
==============================Original Headers============================== 15, 29 -- From Walter.Bobrowski-at-pfizer.com Thu Nov 2 13:39:15 2006 15, 29 -- Received: from gromsgoa01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 15, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA2JdDpG019378 15, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 2 Nov 2006 13:39:14 -0600 15, 29 -- Received: from groamrexc01.amer.pfizer.com (groamrexc01.amer.pfizer.com [172.30.8.168]) 15, 29 -- by gromsgoa01.pfizer.com (8.13.4/8.13.4) with ESMTP id kA2JcvJ0029861; 15, 29 -- Thu, 2 Nov 2006 14:38:57 -0500 15, 29 -- Received: from groamrexc01.amer.pfizer.com ([172.30.8.157]) by groamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.211); 15, 29 -- Thu, 2 Nov 2006 14:39:10 -0500 15, 29 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by groamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.211); 15, 29 -- Thu, 2 Nov 2006 14:39:10 -0500 15, 29 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.7226.0 15, 29 -- Content-class: urn:content-classes:message 15, 29 -- MIME-Version: 1.0 15, 29 -- Content-Type: text/plain; 15, 29 -- charset="US-ASCII" 15, 29 -- Subject: RE: [Microscopy] viaWWW: TEM- EDS on vitrified samples 15, 29 -- Date: Thu, 2 Nov 2006 14:39:10 -0500 15, 29 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D07EEC97E-at-anaamrexm01.amer.pfizer.com} 15, 29 -- X-MS-Has-Attach: 15, 29 -- X-MS-TNEF-Correlator: 15, 29 -- Thread-Topic: [Microscopy] viaWWW: TEM- EDS on vitrified samples 15, 29 -- Thread-Index: Acb+MUFW4fTyUaHnQfG07OhNW/5IdQAhJFIw 15, 29 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 15, 29 -- To: {cervantes-at-bendres.com} , {Microscopy-at-microscopy.com} 15, 29 -- X-OriginalArrivalTime: 02 Nov 2006 19:39:10.0711 (UTC) FILETIME=[919CC070:01C6FEB6] 15, 29 -- X-Proofpoint-Spam-Reason: safe 15, 29 -- Content-Transfer-Encoding: 8bit 15, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA2JdDpG019378 ==============================End of - Headers==============================
When I visited Zeiss in Germany, they had a reasonable size unit that polished small to large (metallurgical) samples using plasma. The unit I think sat on a table. It had a slide to the right cover to insert the specimen, then it would slide back and lock. There was a color flat panel LCD display on the far right of the unit to concoct and recall recipes. This was Zeiss' standard way of preparing and cleaning specimens. It was a very nice unit and was reported to be quite reliable and produced excellent results.
Does anyone know what this system might be? I did contact Heiner but so far, no response.
tnx, gary g.
==============================Original Headers============================== 5, 17 -- From gary-at-gaugler.com Thu Nov 2 15:43:17 2006 5, 17 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 5, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA2LhGi8032617 5, 17 -- for {microscopy-at-microscopy.com} ; Thu, 2 Nov 2006 15:43:16 -0600 5, 17 -- Received: (qmail 17888 invoked from network); 2 Nov 2006 13:43:16 -0800 5, 17 -- Received: by simscan 1.1.0 ppid: 17858, pid: 17883, t: 0.1097s 5, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 5, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 5, 17 -- by qsmtp3 with SMTP; 2 Nov 2006 13:43:16 -0800 5, 17 -- Message-Id: {7.0.1.0.2.20061102133830.024c2d60-at-gaugler.com} 5, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 5, 17 -- Date: Thu, 02 Nov 2006 13:43:09 -0800 5, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 5, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 5, 17 -- Subject: EBSD polishing system 5, 17 -- Mime-Version: 1.0 5, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
} When I visited Zeiss in Germany, they had a reasonable size } unit that polished small to large (metallurgical) samples } using plasma. The unit I think sat on a table. } ... } } Does anyone know what this system might be? I did contact } Heiner but so far, no response.
It may have been something akin to the JEOL SM-09010 "cross section polisher"
Occasionally we encounter situations where, following pretreatments, we achieve successful immunolabeling on resin-embedded thick sectioned samples, but only in distinct areas of the tissue section, where as the rest of the section is unexpectedly unlabelled. What is interesting is that the pattern of (inconsistent) labeling is fairly consistent among all the replicate sections on the glass slide, which randomly dry down in various orientations. At other times, the entire tissue section is labeled successfully.
Has anyone encountered this, and if so, is there an explanation? We're assuming it's a technique issue rather than an issue with either the tissue itself or the resin matrix.
Best regards,
Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
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==============================Original Headers============================== 8, 29 -- From Walter.Bobrowski-at-pfizer.com Fri Nov 3 07:57:47 2006 8, 29 -- Received: from gromsgom01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 8, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA3DvkvA007191 8, 29 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Nov 2006 07:57:46 -0600 8, 29 -- Received: from groamrexc02.amer.pfizer.com (groamrexc02.amer.pfizer.com [172.30.8.169]) 8, 29 -- by gromsgom01.pfizer.com (8.13.4/8.13.4) with ESMTP id kA3DutjV032233 8, 29 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Nov 2006 08:56:55 -0500 8, 29 -- Received: from mopamrexc02.amer.pfizer.com ([170.116.30.68]) by groamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.211); 8, 29 -- Fri, 3 Nov 2006 08:57:45 -0500 8, 29 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 8, 29 -- Fri, 3 Nov 2006 08:57:45 -0500 8, 29 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.7226.0 8, 29 -- Content-class: urn:content-classes:message 8, 29 -- MIME-Version: 1.0 8, 29 -- Content-Type: text/plain; 8, 29 -- charset="US-ASCII" 8, 29 -- Subject: "Spotty" Immunolabeling on Resin-Embedded Samples 8, 29 -- Date: Fri, 3 Nov 2006 08:57:39 -0500 8, 29 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D07EECCF4-at-anaamrexm01.amer.pfizer.com} 8, 29 -- X-MS-Has-Attach: 8, 29 -- X-MS-TNEF-Correlator: 8, 29 -- Thread-Topic: "Spotty" Immunolabeling on Resin-Embedded Samples 8, 29 -- Thread-Index: Acb/UAaEstnS5mjmTXeuIO2rRkS3Pw== 8, 29 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 8, 29 -- To: {Microscopy-at-microscopy.com} 8, 29 -- X-OriginalArrivalTime: 03 Nov 2006 13:57:45.0667 (UTC) FILETIME=[09FCD930:01C6FF50] 8, 29 -- X-Proofpoint-Spam-Reason: safe 8, 29 -- Content-Transfer-Encoding: 8bit 8, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA3DvkvA007191 ==============================End of - Headers==============================
I need to quantify total specific staining on immunolabeled microscope slides. I did this many years ago with an old DOS image analysis program (Jandel's JAVA) using the Beer-Lambert equation to compute the total absorbance on a pixel by pixel basis. Does anybody know if there are plug-ins or macros available to do this with ImageJ? I would prefer to use software with a history of use and citation, if possible. Alternatively, is there a better way to quantify staining?
Ralph Common Michigan State University Dept. of Physiology
==============================Original Headers============================== 3, 24 -- From rcommon-at-msu.edu Fri Nov 3 09:13:08 2006 3, 24 -- Received: from sys19.mail.msu.edu (sys19.mail.msu.edu [35.9.75.119]) 3, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA3FD8SJ030040 3, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 3 Nov 2006 09:13:08 -0600 3, 24 -- Received: from [35.9.122.125] (helo=emlab) 3, 24 -- by sys19.mail.msu.edu with esmtpsa (Exim 4.52 #1) 3, 24 -- (TLSv1:RC4-MD5:128) 3, 24 -- id 1Gg0jE-0004S4-3o 3, 24 -- for Microscopy-at-microscopy.com; Fri, 03 Nov 2006 10:13:08 -0500 3, 24 -- From: "Ralph Common" {rcommon-at-msu.edu} 3, 24 -- To: {Microscopy-at-microscopy.com} 3, 24 -- Subject: Quantifying specific staining 3, 24 -- Date: Fri, 3 Nov 2006 10:14:32 -0500 3, 24 -- Message-ID: {005a01c6ff5a$c4654af0$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 V6.00.2800.1807 3, 24 -- Importance: Normal 3, 24 -- X-Virus: None found by Clam AV ==============================End of - Headers==============================
Its purpose was to polish specimens. Now that I look at the several responses, it must have been an ion mill type of system. It looks similar to the Fishione system in size but the right half of the cover slides to the right to open the chamber. The control console is a flat panel LCD on an articulating arm. As I recall, it used only Ar.
gary g.
At 02:57 AM 11/3/2006, you wrote: } Dear Gary, } } Are you sure it polished as well? I've } seen cleaners (Oxygen plasma to remove surface } grot), but I've never heard of a } polisher/etcher for SEM work. [Although I have } done reactive ion etching of semiconductors using (halogenated) hydrocarbons.] } } Was it anything like the Fishione } instruments 1020? http://www.fischione.com/products/model_1020.asp } } Austin } } P.S. It's very unlikely, but was it was an ion-beam miller? } http://www.gatan.com/specimenprep/691_pips.html } You can do very nice EBSD prep on small areas } using a highly tilted specimen } 80°, low kV } ( {5keV) and a defocused beam. It can also remove surface grot. } } P.P.S. There are other plasma etcher manufacturers, e.g. SPI. } http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml } } ----- Original Message ----- From: {gary-at-gaugler.com} } To: {AuntDaisy-at-gmail.com} } Sent: Thursday, November 02, 2006 9:49 PM } Subject: [Microscopy] EBSD polishing system } } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 7, 23 -- From gary-at-gaugler.com Fri Nov 3 10:32:46 2006 7, 23 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA3GWiTq011063 7, 23 -- for {microscopy-at-microscopy.com} ; Fri, 3 Nov 2006 10:32:45 -0600 7, 23 -- Received: (qmail 3496 invoked from network); 3 Nov 2006 08:32:43 -0800 7, 23 -- Received: by simscan 1.1.0 ppid: 3437, pid: 3494, t: 0.0975s 7, 23 -- scanners: regex: 1.1.0 attach: 1.1.0 7, 23 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 7, 23 -- by qsmtp3 with SMTP; 3 Nov 2006 08:32:43 -0800 7, 23 -- Message-Id: {7.0.1.0.2.20061103082941.02612f20-at-gaugler.com} 7, 23 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 7, 23 -- Date: Fri, 03 Nov 2006 08:32:37 -0800 7, 23 -- To: ADay {auntdaisy-at-gmail.com} 7, 23 -- From: Gary Gaugler {gary-at-gaugler.com} 7, 23 -- Subject: Re: [Microscopy] EBSD polishing system 7, 23 -- Cc: MSA listserver {microscopy-at-microscopy.com} 7, 23 -- In-Reply-To: {005601c6ff36$d08b6130$0100a8c0-at-BigDell} 7, 23 -- References: {200611022149.kA2LnmBJ006384-at-ns.microscopy.com} 7, 23 -- {005601c6ff36$d08b6130$0100a8c0-at-BigDell} 7, 23 -- Mime-Version: 1.0 7, 23 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA3GWiTq011063 ==============================End of - Headers==============================
Gary: I'm wondering if it was an RES100 ion mill by Bal-Tec. I demo'd the RES100 back when it first came out and if my memory is reliable (sometimes not) it had a sliding cover. I don't think they sell the RES100 anymore. I recommend the RES101, the latest generation of that tool, which I have and use almost everyday. I use it for SEM sample clean-up and cross-section delineation. In the USA, Bal-Tec is rep'd by RMC Boeckler. http://www.baltec-rmc.com/cms/index.cfm/path/17933/26002/24159/25864/ PS: there are NO spaces in the URL and I have no financial interest in the company.
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 } ---------------------------------------------------------------------------- } } Its purpose was to polish specimens. Now that I look } at the several responses, it must have been an ion mill } type of system. It looks similar to the Fishione system } in size but the right half of the cover slides to the right } to open the chamber. The control console is a flat panel } LCD on an articulating arm. As I recall, it used only } Ar. } } gary g. } } } } At 02:57 AM 11/3/2006, you wrote: } } } Dear Gary, } } } } Are you sure it polished as well? I've } } seen cleaners (Oxygen plasma to remove surface } } grot), but I've never heard of a } } polisher/etcher for SEM work. [Although I have } } done reactive ion etching of semiconductors using (halogenated) hydrocarbons.] } } } } Was it anything like the Fishione } } instruments 1020? http://www.fischione.com/products/model_1020.asp } } } } Austin } } } } P.S. It's very unlikely, but was it was an ion-beam miller? } } http://www.gatan.com/specimenprep/691_pips.html } } You can do very nice EBSD prep on small areas } } using a highly tilted specimen } 80°, low kV } } ( {5keV) and a defocused beam. It can also remove surface grot. } } } } P.P.S. There are other plasma etcher manufacturers, e.g. SPI. } } http://www.2spi.com/catalog/instruments/plasma-cleaner.shtml } } } } ----- Original Message ----- From: {gary-at-gaugler.com} } } To: {AuntDaisy-at-gmail.com} } } Sent: Thursday, November 02, 2006 9:49 PM } } Subject: [Microscopy] EBSD polishing system } } } } } } } } } } } } ---------------------------------------------------------------------------- } } } 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 Listers: } } } } } } When I visited Zeiss in Germany, they had a reasonable } } } size unit that polished small to large (metallurgical) } } } samples using plasma. The unit I think sat on a table. } } } It had a slide to the right cover to insert the specimen, } } } then it would slide back and lock. There was a color } } } flat panel LCD display on the far right of the unit to } } } concoct and recall recipes. This was Zeiss' standard } } } way of preparing and cleaning specimens. It was a very } } } nice unit and was reported to be quite reliable and produced } } } excellent results. } } } } } } Does anyone know what this system might be? I did } } } contact Heiner but so far, no response. } } } } } } tnx, } } } gary g. } } } } } } } } } ==============================Original Headers============================== } } } 5, 17 -- From gary-at-gaugler.com Thu Nov 2 15:43:17 2006 } } } 5, 17 -- Received: from qsmtp3.mc.surewest.net } } } (qsmtp.mc.surewest.net [66.60.130.145]) } } } 5, 17 -- by ns.microscopy.com } } } (8.12.11.20060308/8.12.8) with SMTP id kA2LhGi8032617 } } } 5, 17 -- for {microscopy-at-microscopy.com} ; Thu, 2 Nov 2006 15:43:16 -0600 } } } 5, 17 -- Received: (qmail 17888 invoked from } } } network); 2 Nov 2006 13:43:16 -0800 } } } 5, 17 -- Received: by simscan 1.1.0 ppid: 17858, pid: 17883, t: 0.1097s } } } 5, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 } } } 5, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) } } } 5, 17 -- by qsmtp3 with SMTP; 2 Nov 2006 13:43:16 -0800 } } } 5, 17 -- Message-Id: {7.0.1.0.2.20061102133830.024c2d60-at-gaugler.com} } } } 5, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } } } 5, 17 -- Date: Thu, 02 Nov 2006 13:43:09 -0800 } } } 5, 17 -- To: MSA listserver {microscopy-at-microscopy.com} } } } 5, 17 -- From: Gary Gaugler {gary-at-gaugler.com} } } } 5, 17 -- Subject: EBSD polishing system } } } 5, 17 -- Mime-Version: 1.0 } } } 5, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } } } ==============================End of - Headers============================== } } } } } } } ==============================Original Headers============================== } 7, 23 -- From gary-at-gaugler.com Fri Nov 3 10:32:46 2006 } 7, 23 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) } 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA3GWiTq011063 } 7, 23 -- for {microscopy-at-microscopy.com} ; Fri, 3 Nov 2006 10:32:45 -0600 } 7, 23 -- Received: (qmail 3496 invoked from network); 3 Nov 2006 08:32:43 -0800 } 7, 23 -- Received: by simscan 1.1.0 ppid: 3437, pid: 3494, t: 0.0975s } 7, 23 -- scanners: regex: 1.1.0 attach: 1.1.0 } 7, 23 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) } 7, 23 -- by qsmtp3 with SMTP; 3 Nov 2006 08:32:43 -0800 } 7, 23 -- Message-Id: {7.0.1.0.2.20061103082941.02612f20-at-gaugler.com} } 7, 23 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 } 7, 23 -- Date: Fri, 03 Nov 2006 08:32:37 -0800 } 7, 23 -- To: ADay {auntdaisy-at-gmail.com} } 7, 23 -- From: Gary Gaugler {gary-at-gaugler.com} } 7, 23 -- Subject: Re: [Microscopy] EBSD polishing system } 7, 23 -- Cc: MSA listserver {microscopy-at-microscopy.com} } 7, 23 -- In-Reply-To: {005601c6ff36$d08b6130$0100a8c0-at-BigDell} } 7, 23 -- References: {200611022149.kA2LnmBJ006384-at-ns.microscopy.com} } 7, 23 -- {005601c6ff36$d08b6130$0100a8c0-at-BigDell} } 7, 23 -- Mime-Version: 1.0 } 7, 23 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed } 7, 23 -- Content-Transfer-Encoding: 8bit } 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA3GWiTq011063 } ==============================End of - Headers============================== } }
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
==============================Original Headers============================== 4, 23 -- From r-holdford-at-ti.com Fri Nov 3 18:08:06 2006 4, 23 -- Received: from calf.ext.ti.com (calf.ext.ti.com [198.47.26.144]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA4086a7031351 4, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 3 Nov 2006 18:08:06 -0600 4, 23 -- Received: from dlep32.itg.ti.com ([157.170.170.70]) 4, 23 -- by calf.ext.ti.com (8.13.7/8.13.7) with ESMTP id kA407UWE005958 4, 23 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO); 4, 23 -- Fri, 3 Nov 2006 18:07:40 -0600 4, 23 -- Received: from [156.117.194.120] (localhost [127.0.0.1]) 4, 23 -- by dlep32.itg.ti.com (8.13.7/8.13.7) with ESMTP id kA407OEl019664; 4, 23 -- Fri, 3 Nov 2006 18:07:24 -0600 (CST) 4, 23 -- Message-ID: {454BD9BC.6060201-at-ti.com} 4, 23 -- Date: Fri, 03 Nov 2006 18:07:24 -0600 4, 23 -- From: Becky Holdford {r-holdford-at-ti.com} 4, 23 -- Organization: SC Packaging Development -- FA Development 4, 23 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 4, 23 -- MIME-Version: 1.0 4, 23 -- To: gary-at-gaugler.com, MSA ListServer {Microscopy-at-MSA.Microscopy.Com} 4, 23 -- Subject: Re: [Microscopy] Re: EBSD polishing system 4, 23 -- References: {200611031633.kA3GXMI2011396-at-ns.microscopy.com} 4, 23 -- In-Reply-To: {200611031633.kA3GXMI2011396-at-ns.microscopy.com} 4, 23 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 23 -- 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 dgmorgan-at-ucdavis.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: dgmorgan-at-ucdavis.edu Name: David Morgan
Organization: UC Davis
Title-Subject: [Filtered] Tantalum foils
Question: Hi,
I find myself in need of a tantalum foil suitable for TEM and since I am just getting into the field of materials science, I am hoping that this sort of sample is commercially available. Are such things available and can anyone suggest a vendor? Thanks in advance.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both elchem33-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: elchem33-at-gmail.com Name: Marco Puzi
Organization: Dep of Chemistry and Biochemistry, Univ Split, Croatia
Title-Subject: [Filtered] TEM alignment problem
Question: Hi!
During the alignment of our Jeol JEM-200CX TEM something has gone wrong. I suppose that user by mistake used the gun/beam tilt controls instead the shift (trans) controls, or he altered the sequence for gun and condenser alignment; after that, it seems that he used other controls (deflector coils X and Y corrector/compensator, beam displacement compensating coil... I even suspect that he mechanically moved the intermediate and projector lens) in attempt to align the microscope. As a result, the microscope is terribly misaligned and I can't seem to get it back in the aligned state following the alignment procedure given in Jeol manual. After the gun and condenser alignment steps everything seems fine (beam is centered on the screen and expands uniformly). However, when cond alignment wobbler is turned on, the beam separates into two spots, and one spot is blurred to aprox. 1 cm in diameter, while other remains focused! Also, when specimen is placed in microscope, image moves badly during focusing and becomes blurred during the change of illumination. I suppose that all this effects are due to the tilted beam.
Does anybody know the alignment procedure that works in similar situations?
Also, I will be grateful if anyone could explain me what beam displacement compensating coils (located between condenser and objective lens, just below beam deflector coils) exactly do (this topic is not covered in manual).
You can buy sheets of Tantalum metal at Goodfellow They sell small quantities with high purity for research.
Here is the link.
http://www.goodfellow.com/
Just search for tantalum foils.
These are generally suitable for you to electropolish to make TEM samples. Warning I believe anumber of the polishing solutions tend to use HF, nasty stuff. If your new to TEM sample method make sure you know the hazards of HF.
Nestor Your Friendly Neighborhood SysOp
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==============================Original Headers============================== 15, 13 -- From zaluzec-at-microscopy.com Sat Nov 4 12:44:14 2006 15, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 15, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA4IiEN1014959; 15, 13 -- Sat, 4 Nov 2006 12:44:14 -0600 15, 13 -- Mime-Version: 1.0 15, 13 -- Message-Id: {p06110402c1728d62bb1d-at-[206.69.208.22]} 15, 13 -- In-Reply-To: {200611041829.kA4ITo4u026249-at-ns.microscopy.com} 15, 13 -- References: {200611041829.kA4ITo4u026249-at-ns.microscopy.com} 15, 13 -- Date: Sat, 4 Nov 2006 12:44:14 -0600 15, 13 -- To: dgmorgan-at-ucdavis.edu, microscopy-at-microscopy.com 15, 13 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 15, 13 -- Subject: Re: [Microscopy] viaWWW: Tantalum foils 15, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
D. Morgan asked: ============================================= I find myself in need of a tantalum foil suitable for TEM and since I am just getting into the field of materials science, I am hoping that this sort of sample is commercially available. Are such things available and can anyone suggest a vendor? Thanks in advance. ============================================= If you are talking about a tantalum foil on a TEM grid, see URL http://www.2spi.com/catalog/standards/aem.shtml
This is a standard foil product produced by SPI Supplies.
Disclaimer: SPI Supplies is the manufacturer of this foil sample.
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
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 11, 30 -- From cgarber-at-2spi.com Sat Nov 4 14:56:41 2006 11, 30 -- Received: from s-utl01-atpop.stsn.net (s-utl01-atpop.stsn.net [72.254.128.201]) 11, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA4KueDT003474 11, 30 -- for {microscopy-at-msa.microscopy.com} ; Sat, 4 Nov 2006 14:56:40 -0600 11, 30 -- Received: from s-utl01-atpop.stsn.net ([127.0.0.1]) 11, 30 -- by s-utl01-atpop.stsn.net (SMSSMTP 4.1.2.20) with SMTP id M2006110415563923259 11, 30 -- for {microscopy-at-msa.microscopy.com} ; Sat, 04 Nov 2006 15:56:39 -0500 11, 30 -- X-Spam-Status: No, hits=0.0 required=9.9 11, 30 -- tests=ALL_TRUSTED: -2.867,AWL: 0.139,BAYES_00: -1.665, 11, 30 -- SARE_RECV_ADDR: 0.027 11, 30 -- X-Spam-Level: 11, 30 -- Received: from ibm1x23g2abfyg ([10.1.154.1]) 11, 30 -- by s-utl01-atpop.stsn.net 11, 30 -- for microscopy-at-msa.microscopy.com; 11, 30 -- Sat, 4 Nov 2006 15:56:38 -0500 11, 30 -- Message-ID: {001d01c70053$8daa43e0$019a010a-at-ibm1x23g2abfyg} 11, 30 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 11, 30 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 11, 30 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 11, 30 -- Subject: Tantalum foil on a grid 11, 30 -- Date: Sat, 4 Nov 2006 15:55:15 -0500 11, 30 -- MIME-Version: 1.0 11, 30 -- Content-Type: text/plain; 11, 30 -- charset="Windows-1252" 11, 30 -- X-Priority: 3 11, 30 -- X-MSMail-Priority: Normal 11, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 11, 30 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 11, 30 -- Content-Transfer-Encoding: 8bit 11, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA4KueDT003474 ==============================End of - Headers==============================
Also available from other electron microscope supply companies. You might want to do a price check.
Ted Dunn The EMscope Company Ltd. Thailand
D. Morgan asked: ============================================= I find myself in need of a tantalum foil suitable for TEM
--- cgarber-at-2spi.com wrote:
============================================= } If you are talking about a tantalum foil on a TEM } grid, see URL } http://www.2spi.com/catalog/standards/aem.shtml } } This is a standard foil product produced by SPI } Supplies. } } Disclaimer: SPI Supplies is the manufacturer of } this foil sample. } } 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 } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== } } } } } } } ==============================Original } Headers============================== } 11, 30 -- From cgarber-at-2spi.com Sat Nov 4 14:56:41 } 2006 } 11, 30 -- Received: from s-utl01-atpop.stsn.net } (s-utl01-atpop.stsn.net [72.254.128.201]) } 11, 30 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } kA4KueDT003474 } 11, 30 -- for {microscopy-at-msa.microscopy.com} ; Sat, } 4 Nov 2006 14:56:40 -0600 } 11, 30 -- Received: from s-utl01-atpop.stsn.net } ([127.0.0.1]) } 11, 30 -- by s-utl01-atpop.stsn.net (SMSSMTP } 4.1.2.20) with SMTP id M2006110415563923259 } 11, 30 -- for {microscopy-at-msa.microscopy.com} ; Sat, } 04 Nov 2006 15:56:39 -0500 } 11, 30 -- X-Spam-Status: No, hits=0.0 required=9.9 } 11, 30 -- tests=ALL_TRUSTED: -2.867,AWL: } 0.139,BAYES_00: -1.665, } 11, 30 -- SARE_RECV_ADDR: 0.027 } 11, 30 -- X-Spam-Level: } 11, 30 -- Received: from ibm1x23g2abfyg } ([10.1.154.1]) } 11, 30 -- by s-utl01-atpop.stsn.net } 11, 30 -- for microscopy-at-msa.microscopy.com; } 11, 30 -- Sat, 4 Nov 2006 15:56:38 -0500 } 11, 30 -- Message-ID: } {001d01c70053$8daa43e0$019a010a-at-ibm1x23g2abfyg} } 11, 30 -- Reply-To: "Garber, Charles A." } {cgarber-at-2spi.com} } 11, 30 -- From: "Garber, Charles A." } {cgarber-at-2spi.com} } 11, 30 -- To: "Microscopy Listserver" } {microscopy-at-msa.microscopy.com} } 11, 30 -- Subject: Tantalum foil on a grid } 11, 30 -- Date: Sat, 4 Nov 2006 15:55:15 -0500 } 11, 30 -- MIME-Version: 1.0 } 11, 30 -- Content-Type: text/plain; } 11, 30 -- charset="Windows-1252" } 11, 30 -- X-Priority: 3 } 11, 30 -- X-MSMail-Priority: Normal } 11, 30 -- X-Mailer: Microsoft Outlook Express } 6.00.2900.2869 } 11, 30 -- X-MimeOLE: Produced By Microsoft MimeOLE } V6.00.2900.2962 } 11, 30 -- Content-Transfer-Encoding: 8bit } 11, 30 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id } kA4KueDT003474 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 10, 19 -- From drteddunne-at-yahoo.com Sat Nov 4 23:37:29 2006 10, 19 -- Received: from web33413.mail.mud.yahoo.com (web33413.mail.mud.yahoo.com [68.142.206.145]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA55bTi5003526 10, 19 -- for {microscopy-at-microscopy.com} ; Sat, 4 Nov 2006 23:37:29 -0600 10, 19 -- Received: (qmail 75617 invoked by uid 60001); 5 Nov 2006 05:37:28 -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=yF+sNbUCO3kL7voiLV9qllMzj/R1qXJ8jPKtZGl+Y2lJdVeore8q+rD7mdjLjZIyeJnzqUxlPdmh4pqnRfZtQImH8hSQqePk1+aOWUfaem+JYntpIfzKtQpEG3ggsE7VoTinI09FLKnZby9QLTcjmcikwVVAOtqfndKzTzZFPxw= ; 10, 19 -- Message-ID: {20061105053728.75615.qmail-at-web33413.mail.mud.yahoo.com} 10, 19 -- Received: from [203.190.250.105] by web33413.mail.mud.yahoo.com via HTTP; Sat, 04 Nov 2006 21:37:28 PST 10, 19 -- Date: Sat, 4 Nov 2006 21:37:28 -0800 (PST) 10, 19 -- From: ted dunn {drteddunne-at-yahoo.com} 10, 19 -- Subject: Re: [Microscopy] Tantalum foil on a grid 10, 19 -- To: microscopy-at-microscopy.com 10, 19 -- In-Reply-To: {200611042059.kA4Kxi21008928-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==============================
However, things may not be as bad as you think. You say that the user may have adjusted the IL and PL mechanical alignments, that is possible but in over 35 years of looking after EMs in government and university labs I've never met a user who adjusted the mechanical alignment. You will soon tell if they have because the Mag images and Diff patterns will not be centered when you go down in mag or camera length. Yes - go down not up. If the image is not quite centered at 1K it will be a long way out at 100K but if it's a bit out at 100K it will be centered at 1K.
So to your Beam Compensation controls. You have two sets of 4 four coils one below the other, each set has 2 X coils and 2 Y coils with X and Y set at 90deg to each other. These are often called Beam Shift (top set) and Beam Tilt (lower set). However both sets are used to give shift and tilt. Imagine you have a beam running down the column and you pass a current through the top X set, this will deflect the beam through an angle - alpha. If you then pass an equal current in the opposite direction through the X coils in the lower set the beam will be deflected through an angle, alpha, but in the opposite direction. The beam will now be parallel with the original direction but moved (shifted) a little. Now apply a higher current through the lower set and the beam will be deflected through a greater angle. If higher current is set correctly the beam will pass through the same point in the specimen as the undefleted beam would have but now it is at an angle (beta) to the original beam. This is pure tilt, increase the current through both the upper and lower sets in the same ratio and the angle (beta) changes. The beam shift control adjusts the coils in the correct ratio to shift and the beam tilt control adjusts them in the correct ratio to tilt. This is a whole lot easier to understand if you draw it out.
With the beam compensators you are setting up pure tilt onto the specimen. The microscope wobbles the beam and you adjust the ratio until both spots coincide. Under this condition when you tilt the beam (eg. for dark field imaging) it does not move off the area of interest.
Now what about your problem - I think that the distortion you see is caused by the beam being deflected too far from the optic axis. First (assuming you have a side entry 200CX) make sure the specimen is at eucentric height (image does not move when you tilt the specimen rod) then focus the specimen. If you have a top entry 200CX then set the objective current to the correct value. Next set the beam tilts to their mid positions, that is when both indicator arrows are lit or when it changes from one to the other. If the beam tilt is a long way out the image will move a lot as you adjust the objective lens current.
Now try the beam compensation again. I suspect that either the objective lens current is wrong or the beam tilt is a long way out to begin with.
Of course it is always possible that the user has adjusted the mechanical alignments or that lenses, deflectors or HT are not working properly but in my experience that is less likely.
I hope this helps.
Good luck, Ron
In message {200611041839.kA4IdG3M012999-at-ns.microscopy.com} elchem33-at-gmail.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 } ---------------------------------------------------------------------------- } } 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 elchem33-at-gmail.com as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: elchem33-at-gmail.com } Name: Marco Puzi } } Organization: Dep of Chemistry and Biochemistry, Univ Split, Croatia } } Title-Subject: [Filtered] TEM alignment problem } } Question: Hi! } } During the alignment of our Jeol JEM-200CX TEM something has gone wrong. I suppose that user by mistake used the gun/beam tilt controls instead the shift (trans) controls, or he altered the sequence for gun and condenser alignment; after that, it seems that he used other controls (deflector coils X and Y corrector/compensator, beam displacement compensating coil... I even suspect that he mechanically moved the intermediate and projector lens) in attempt to align the microscope. As a result, the microscope is terribly misaligned and I can't seem to get it back in the aligned state following the alignment procedure given in Jeol manual. After the gun and condenser alignment steps everything seems fine (beam is centered on the screen and expands uniformly). However, when cond alignment wobbler is turned on, the beam separates into two spots, and one spot is blurred to aprox. 1 cm in diameter, while other remains focused! Also, when specimen is placed in microscope, image moves ! } badly during focusing and becomes blurred during the change of illumination. I suppose that all this effects are due to the tilted beam. } } Does anybody know the alignment procedure that works in similar situations? } } Also, I will be grateful if anyone could explain me what beam displacement compensating coils (located between condenser and objective lens, just below beam deflector coils) exactly do (this topic is not covered in manual). } } Thanx in advance. } Marco } } } --------------------------------------------------------------------------- } } } ==============================Original Headers============================== } 12, 14 -- From zaluzec-at-microscopy.com Sat Nov 4 12:31:07 2006 } 12, 14 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) } 12, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA4IV6TA027303 } 12, 14 -- for {microscopy-at-microscopy.com} ; Sat, 4 Nov 2006 12:31:07 -0600 } 12, 14 -- Mime-Version: 1.0 } 12, 14 -- X-Sender: (Unverified) } 12, 14 -- Message-Id: {p06110401c1728ca58ed7-at-[206.69.208.22]} } 12, 14 -- Date: Sat, 4 Nov 2006 12:31:07 -0600 } 12, 14 -- To: microscopy-at-microscopy.com } 12, 14 -- From: elchem33-at-gmail.com (by way of MicroscopyListserver) } 12, 14 -- Subject: viaWWW: TEM alignment problem Help needed } 12, 14 -- Content-Type: text/plain; charset="us-ascii" } 12, 14 -- Content-Transfer-Encoding: 8bit } 12, 14 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA4IV6TA027303 } ==============================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============================== 14, 24 -- From ron.doole-at-materials.ox.ac.uk Sun Nov 5 15:48:48 2006 14, 24 -- Received: from relay0.mail.ox.ac.uk (relay0.mail.ox.ac.uk [129.67.1.161]) 14, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA5Lml3j013170 14, 24 -- for {microscopy-at-msa.microscopy.com} ; Sun, 5 Nov 2006 15:48:47 -0600 14, 24 -- Received: from webmail218.herald.ox.ac.uk ([163.1.0.218]) 14, 24 -- by relay0.mail.ox.ac.uk with esmtp (Exim 4.62) 14, 24 -- (envelope-from {ron.doole-at-materials.ox.ac.uk} ) 14, 24 -- id 1GgprC-0007IN-37; Sun, 05 Nov 2006 21:48:47 +0000 14, 24 -- Received: by webmail218.herald.ox.ac.uk (Postfix, from userid 33) 14, 24 -- id E75D65A078; Sun, 5 Nov 2006 21:48:46 +0000 (GMT) 14, 24 -- Content-Type: text/plain 14, 24 -- Content-Disposition: inline 14, 24 -- Content-Transfer-Encoding: 7bit 14, 24 -- MIME-Version: 1.0 14, 24 -- X-Mailer: MIME-tools 5.417 (Entity 5.417) 14, 24 -- From: Ron Doole {ron.doole-at-materials.ox.ac.uk} 14, 24 -- Date: Sun, 05 Nov 2006 21:48:46 +0000 14, 24 -- To: elchem33-at-gmail.com 14, 24 -- Cc: microscopy-at-msa.microscopy.com 14, 24 -- Subject: Re: [Microscopy] viaWWW: TEM alignment problem Help needed 14, 24 -- In-Reply-To: {200611041839.kA4IdG3M012999-at-ns.microscopy.com} 14, 24 -- X-Webmail-Sender: rdoole 14, 24 -- X-Webmail-Originating-Ip: 86.134.92.244 14, 24 -- Message-Id: {20061105214846.E75D65A078-at-webmail218.herald.ox.ac.uk} ==============================End of - Headers==============================
We shall be having an Open Day for students next week and one of the highlights this time is 'showcasing' our newly acquired SEM unit. Any ideas on how I could present the new toy to them and catch their attention? Posters or movies you could share perhaps? =)
Our unit is a FEI Quanta 200, by the way.
Thanks and best regards,
Melina L. Miralles PGSc Laboratory Technician The Petroleum Institute Tel: 02-5085497 (Office) Tel: 02-5085539/5481 (Lab) Fax : 02-5085423
==============================Original Headers============================== 9, 27 -- From mmiralles-at-pi.ac.ae Mon Nov 6 01:26:42 2006 9, 27 -- Received: from mx1.pi.ac.ae (mx1.pi.ac.ae [213.42.148.228]) 9, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA67QdON002122 9, 27 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 01:26:41 -0600 9, 27 -- Received: from pi-exf.pi.ac.ae ([192.168.2.11]) 9, 27 -- by mx1.pi.ac.ae with ESMTP; 06 Nov 2006 11:29:33 +0400 9, 27 -- X-IronPort-AV: i="4.09,390,1157313600"; 9, 27 -- d="scan'208"; a="829514:sNHT21947008" 9, 27 -- Received: from pi-exm.PI.AC.AE ([10.248.1.18]) by PI-EXF.PI.AC.AE with Microsoft SMTPSVC(6.0.3790.1830); 9, 27 -- Mon, 6 Nov 2006 11:26:23 +0400 9, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 27 -- Content-class: urn:content-classes:message 9, 27 -- MIME-Version: 1.0 9, 27 -- Content-Type: text/plain; 9, 27 -- charset="us-ascii" 9, 27 -- Subject: Open Day Ideas?? 9, 27 -- Date: Mon, 6 Nov 2006 11:26:23 +0400 9, 27 -- Message-ID: {D5603421C6303A46883F87E7968F285B01F69E28-at-pi-exm.PI.AC.AE} 9, 27 -- X-MS-Has-Attach: 9, 27 -- X-MS-TNEF-Correlator: 9, 27 -- Thread-Topic: Open Day Ideas?? 9, 27 -- Thread-Index: AccBdN0FLsnEqkD4SHGdRuMNTtUBeQ== 9, 27 -- From: "Melina Miralles" {mmiralles-at-pi.ac.ae} 9, 27 -- To: {Microscopy-at-microscopy.com} 9, 27 -- X-OriginalArrivalTime: 06 Nov 2006 07:26:23.0950 (UTC) FILETIME=[DD086EE0:01C70174] 9, 27 -- Content-Transfer-Encoding: 8bit 9, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA67QdON002122 ==============================End of - Headers==============================
Hello all, we have following trouble: "Panasonic Lithium battery BR-2/3A 3V" which are used for battery back up in Philips CM12 electron microscope were exhausted after one year of use. This occurred for the second time. Is this normal ? Previously we had to replace them after several years but now we have to use the third set of batteries in two years. Has anybody any suggestion or hint? Thanking you in advance. Oldrich
----------------------------------------------- Oldrich Benada Institute of Microbiology, Acad. Sci. CR Videnska 1083 142 20 Prague 4 Czech Republic
==============================Original Headers============================== 5, 20 -- From benada-at-biomed.cas.cz Mon Nov 6 04:14:43 2006 5, 20 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA6AEglN016074 5, 20 -- for {microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 04:14:43 -0600 5, 20 -- Received: from [147.231.44.101] (u117ob.mbu.cas.cz [147.231.44.101]) 5, 20 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id kA6AEKDt010340 5, 20 -- for {microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 11:14:20 +0100 (CET) 5, 20 -- From: benada-at-biomed.cas.cz 5, 20 -- To: microscopy-at-microscopy.com 5, 20 -- Date: Mon, 06 Nov 2006 11:14:39 +0100 5, 20 -- MIME-Version: 1.0 5, 20 -- Subject: Battery back up trouble 5, 20 -- Message-ID: {454F191F.4142.2BF28C-at-benada.biomed.cas.cz} 5, 20 -- Priority: normal 5, 20 -- X-mailer: Pegasus Mail for Windows (4.41) 5, 20 -- Content-type: text/plain; charset=US-ASCII 5, 20 -- Content-transfer-encoding: 7BIT 5, 20 -- Content-description: Mail message body 5, 20 -- X-Antivirus: avast! (VPS 0645-4, 03.11.2006), Outbound message 5, 20 -- X-Antivirus-Status: Clean ==============================End of - Headers==============================
I used a 1978 vintage Edwards 306A evaporator. I found a local microscopist with some 1980s E306A documentation and got some copies made. I have repaired or reconditioned at least three different evaporators over the years including an E306.
I believe CB1 and CB2 are 8 amp ganged double pole thermal circuit breakers that are manually reset and latched closed. This ganging on CB2 is set up so that tripping either CB2-8A -OR- CB2-I/LOCK will cut off power through the two ILC switch contacts (shown in diagrams) and that are controlled (not shown) by the ILC "box". The ITC box is a contactor relay coil. The only normal way to trip the CB2-8A half is to have a current over 8 amps. For example, the RP motor stalls out and the current rises. Breaking circuit continuity near CB2-8A or shorting across that half of CB2 will not trip CB2-I/LOCK. CB2-I/LOCK holds an electrical latch on the ILC "box" contactor coil and that keeps the power turned on to the E306 through both sets of contacts labeled ITC. Since that "relay" coil controls the ITC contacts, CB2-I/LOCK is a latched power supply safety interlock circuit switch. The shunt jumper above the ITC box is shown as an unlabeled wire between terminals with wires 2 and 8 on them in the terminal block diagram. The shunt is not shown as an interlock switch. The analysis of CB2-I/LOCK failure was sent separately. Basically any open in the ITC box branch circuit between wires 1 and 4 shuts off power. Any failure in the RP, DP, or Pirani section that causes a higher current draw through CB2-8A will shut down the power circuits. CB2-8A could be damaged and tripping out at a lower current, however. Monitor the current with a clamp-on ammeter to see how close to 8 amps it is running.
You asked about terms. These may be of general interest to E306A owners. AAV-Air Admit Valve HT acc- High Tension Accessory or voltage supply. (Shown as a dashed line rectangular box near the step down transformer, is located between wires 30 & 14, and not labeled as HT in some diagrams.) LTC- Low Tension Contactor. CB2 I/LOCK- InterLock branch of ganged Circuit Breaker two ILC box- InterLock Contactor unit. This is a double pole 230 VAC line voltage detection relay and is latched ON by pushing the CB2 reset. Circle X's- Neon indicator lights.
You said, "the auxiliary contacts of 8-Amp circuit breaker CB2 supply mains power, through a few interlock switches, to a rectangular box labeled 'ILC'." This might mean, "the auxiliary contacts (CB2-I/LOCK) of the dual 8-Amp circuit breaker CB2, supply mains power through an interlock switch (shunt jumper or link above the ILC box) to a rectangular box labeled 'ILC'. What is this ITC thing?" It's a contactor relay coil.
The ILC box should have a dashed line that goes from it to the dashed line that connects the two ganged ILC switch contacts in the main 230 VAC power line feeders. This ILC dashed line would connect near the words MAINS ON which is really a label for the MAINS neon indicator light on the top user's panel.
CB1-8A supplies power through "a few interlock switches", some door interlock switches and to a rectangular box labeled 'LTC' and another shunt. So what is this LTC thing? It is a contactor or coil that operates the LTC switch contacts at the far right, IMO. You turn on LT and those contacts close. That's another missing dashed line to signify remote control of relay contacts.
You mentioned carbon rod evaporations for a reason. I and others had problems on the 306A Edwards with the carbon rod holder too. Check all the connections and clean them, if that is also a problem. Clean the inside surfaces where the rods go. Most of these older stand alone evaporators are bulletproof and still valuable. I restored a second evaporator unit and converted it to the ball bearing operated LADD dual carbon rod and tungsten basket system. Earlier this year I was studying the theory of side emissions of electrons from heated tungsten wires and nichrome wires reputed to cause the false peak. When the wires failed to continue heating, it was always the tightness of the connections at the W wires that were the problem. I don't like to torque things too tight. My prolonged heating and "looser" mechanical connection caused contact failures at the set screws during this prolonged heating. It was not the fault of the LADD unit or my home-made tungsten hairpin holder. (I hate fishing out a slightly bent, jammed and broken W wire from a tiny hole in a wire holder in my garage.) I suspect the same type of contact connection problems for the Edwards carbon rod clamping collars. We really had to torque them very tight.
The rating on CB2-I/LOCK should not be that important but is probably 8 amps at 230 VAC or higher. The ratings should be printed on the items CB2, CB1, and the ILC contactor itself. I am not sure if CB2 is a slow blow circuit breaker or not. I'd call Edwards for a new CB2 unit.
HTH,
Paul
At 01:23 PM 11/1/06 -0600, you wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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Next WEEK? Well, maybe you can be ready for next year. Microscopy Today has David Scharf's posters, and Dennis Kunkel has both a nice poster (see his website) and a really excellent book, "Hidden Worlds". Elaine Humphrey has 3 delightful 3-D (red-green) books titled "Extreme 3-D: Your Body, Scary Bugs, & Weird Animals". You can find info about the books on the MICRO website (URL below). MICRO's site also has a long hotlinked list of websites that students can explore after their visit. If you want something to print & scatter around your walls, go to MICRO's collection of quotes about microscopy.
Caroline -- Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.microscopy.org/ProjectMICRO
==============================Original Headers============================== 3, 18 -- From schooley-at-mcn.org Mon Nov 6 10:55:16 2006 3, 18 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 3, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA6GtFaT012561 3, 18 -- for {microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 10:55:16 -0600 3, 18 -- Received: from [66.52.138.44] (helo=[10.0.1.2]) 3, 18 -- by dns4.mcn.org with esmtpa (Exim 4.60) 3, 18 -- (envelope-from {schooley-at-mcn.org} ) 3, 18 -- id J8BJ01-000BW6-4C; Mon, 06 Nov 2006 08:55:14 -0800 3, 18 -- Mime-Version: 1.0 3, 18 -- Message-Id: {a06200700c175135ef7c4-at-[10.0.1.2]} 3, 18 -- In-Reply-To: {200611060733.kA67XouC010743-at-ns.microscopy.com} 3, 18 -- References: {200611060733.kA67XouC010743-at-ns.microscopy.com} 3, 18 -- Date: Mon, 6 Nov 2006 08:47:15 -0800 3, 18 -- To: mmiralles-at-pi.ac.ae 3, 18 -- From: Caroline Schooley {schooley-at-mcn.org} 3, 18 -- Subject: Re: [Microscopy] Open Day Ideas?? 3, 18 -- Cc: microscopy-at-microscopy.com 3, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Once again, I have been dipping in to "The New Science of Strong Materials" by J. E. Gordon. For those of you who do not know, this is an absolutely wonderful book that tells at a non-technical level what materials science is all about. Thoroughly recommended if you have not read it already.
Gordon says that, in ancient Egypt, mummy cases were made by making papier mâché with papyrus. Some of the papyrus will have writing on it that might be important. On the other hand, no one would want to break up the case of a famous mummy to get pieces of papyrus that might have nothing more on them than somebody's shopping list.
I would have thought that, by now, there ought to be some form of imaging that would let us read what is on the papyrus while it is still one of many layers in the unharmed case of the mummy. I would be pleased to hear either that it has already been done, or that someone is trying to do it.
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============================== 7, 24 -- From jae5-at-lehigh.edu Mon Nov 6 13:29:32 2006 7, 24 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 7, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA6JTVIf026459 7, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 6 Nov 2006 13:29:31 -0600 7, 24 -- Received: from [127.0.0.1] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 7, 24 -- (authenticated bits=0) 7, 24 -- by rain.CC.Lehigh.EDU (8.13.8/8.13.8) with ESMTP id kA6JTO5W027395 7, 24 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 7, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 6 Nov 2006 14:29:29 -0500 7, 24 -- Message-ID: {454F8D14.1010401-at-lehigh.edu} 7, 24 -- Date: Mon, 06 Nov 2006 14:29:24 -0500 7, 24 -- From: Alwyn Eades {jae5-at-lehigh.edu} 7, 24 -- Organization: Lehigh University 7, 24 -- User-Agent: Thunderbird 1.5.0.7 (Windows/20060909) 7, 24 -- MIME-Version: 1.0 7, 24 -- To: MSA listserver {Microscopy-at-MSA.Microscopy.Com} 7, 24 -- Subject: Reading inside a solid object 7, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 24 -- Content-Transfer-Encoding: 8bit 7, 24 -- X-Virus-Scanned: ClamAV version 0.88.6, clamav-milter version 0.88.6 on rain.CC.Lehigh.EDU 7, 24 -- X-Virus-Status: Clean 7, 24 -- X-Spam-Status: No, score=-1.4 required=5.0 tests=ALL_TRUSTED 7, 24 -- autolearn=disabled version=3.1.7 7, 24 -- X-Spam-Checker-Version: SpamAssassin 3.1.7 (2006-10-05) on rain.CC.Lehigh.EDU ==============================End of - Headers==============================
I remember someone posted a message last year asking for help with the low contrast problem he was having on his cultured cells. He stated, after receiving a number of replies, that he has been using the same protocol for years and never had problem until then. I have heard a few other people reporting the same thing to me. Personally, I had an episode like that years ago, but then resolved the problem by using freshly made osmium.
Right now I am experiencing the same problem again, and the problem persisted even when I used newly ordered osmium solution (4% aqueous solution). I also used very short dehydration time to minimize potential lipid lose, but no visible improvement in contrast.
I still think this problem has something to do with osmium. But I would like to see if anyone out there has any new insight.
Thank you in advance.
Hong
Emory SOM EM
==============================Original Headers============================== 10, 25 -- From hyi-at-emory.edu Mon Nov 6 14:44:39 2006 10, 25 -- Received: from nemausa.cc.emory.edu (nemausa.cc.emory.edu [170.140.8.220]) 10, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA6KicfL006121 10, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 14:44:39 -0600 10, 25 -- Received: from nemausa (nemausa [170.140.8.220]) 10, 25 -- by nemausa.cc.emory.edu (8.13.8/8.13.8) with ESMTP id kA6Kicf6019792 10, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 15:44:38 -0500 (EST) 10, 25 -- Received: from nemausa (unknown [127.0.0.1]) 10, 25 -- by nemausa (Symantec Mail Security) with ESMTP id 632A824F6 10, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 15:44:38 -0500 (EST) 10, 25 -- X-AuditID: aa8c08dc-000000090000171d-06-454f9eb6747b 10, 25 -- Received: from [170.140.233.153] (dhcp233153.wmb.emory.edu [170.140.233.153]) 10, 25 -- by nemausa (Symantec Mail Security) with ESMTP id 326BF24F0 10, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 15:44:38 -0500 (EST) 10, 25 -- Mime-Version: 1.0 (Apple Message framework v622) 10, 25 -- Message-Id: {d6203d1f421b9c295156fd4808ea80b7-at-emory.edu} 10, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 25 -- To: Microscopy-at-microscopy.com 10, 25 -- From: Hong Yi {hyi-at-emory.edu} 10, 25 -- Subject: (Microscopy) Low contrast 10, 25 -- Date: Mon, 6 Nov 2006 15:44:33 -0500 10, 25 -- X-Mailer: Apple Mail (2.622) 10, 25 -- X-Brightmail-Tracker: AAAAAA== 10, 25 -- Content-Transfer-Encoding: 8bit 10, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA6KicfL006121 ==============================End of - Headers==============================
Listers, I have just been given a Nanotech Plasmaprep 100 "Plasma Chemistry Unit" that may or may not be functional. It looks to be complete other than a vacuum pump. Does anyone have a manual or wiring diagrams for this unit? Actually, even someone's operating procedures would be better than nothing. Thanks for helping. Kim
-- Kim Rensing Ph.D. Manager, Microscopy and Imaging Facility
University of Calgary, Health Sciences Centre B129 - 3330 Hospital Drive NW Calgary, AB, Canada T2N 4N1
403-220-3488 krensing-at-ucalgary.ca
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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 klangwor-at-uoregon.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: klangwor-at-uoregon.edu Name: Kurt Langworhty
Organization: Univeristy of Oregon
Title-Subject: [Filtered] Surplus Equipment
Question: We are consolidating our confocal and EM labs to prepare for the construction of our Integrated Sciences Building.
I need to donate/sell some equipment to make room in the lab. I'm trying to find homes for:
Varian VE-10 Vacuum Evaporator
Hughes Laser Defractometer (with laser power supply included)
7 boxes of 4x5in Polaroid 55 SEM film (20/box)
Possibly more to come...
Any interested parties should contact me by email (klangwor-at-uoregon.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 zhang.zaoli-at-gmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: zhang.zaoli-at-gmail.com Name: Zaoli
Organization: Ulm univsersity
Title-Subject: [Filtered] TEM specimen preparation about Li compound
Question: Hi,
I am going to carry out the TEM investigation on Li compound single crystals, i.e. LiCoO, or LiFePO4 etc .. I am afraid that routine TEM sample preparation may bring the damage or remove active Li atoms from the sample. does anyone have experience or suggestions on how to prepare TEM samples of such Li compound crystals ? and what kind of damgage could be introduced if a routine TEM sample preparation used ?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pavi_micro-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, November 6, 2006 at 23:57:52 ---------------------------------------------------------------------------
As a material science characterizations engineer with 5 years of work experience, I am searching for a SEM / TEM / SIMS / X-Ray microtomography position, in a laboratory or industrial lab. I have good skills in Electron Microscopy (materials analysis) and in X-Ray microtomography systems. Please ask me (mail adress : samuel_meulenyzer-at-yahoo.fr) my Curriculum Vitae and more details if you are intersted.
As an european citizen, I am free to work anywhere.
Thanks very much.
Samuel Meulenyzer
___________________________________________________________________________ Découvrez une nouvelle façon d'obtenir des réponses à toutes vos questions ! Profitez des connaissances, des opinions et des expériences des internautes sur Yahoo! Questions/Réponses http://fr.answers.yahoo.com
==============================Original Headers============================== 11, 19 -- From samuel_meulenyzer-at-yahoo.fr Tue Nov 7 08:43:17 2006 11, 19 -- Received: from web27901.mail.ukl.yahoo.com (web27901.mail.ukl.yahoo.com [217.146.182.51]) 11, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA7EhGbd025238 11, 19 -- for {Microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 08:43:17 -0600 11, 19 -- Received: (qmail 85797 invoked by uid 60001); 7 Nov 2006 14:43:15 -0000 11, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 19 -- s=s1024; d=yahoo.fr; 11, 19 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 11, 19 -- b=zRvrQQoqkGTTnTqE7mh4hC90U/MgIPSjRyIVjf62AWYvMhljB3KdhONZG3DqbzuTvzHQreEXPeNpxIQ6yd/Z+8436Fx1uR8Q63DLwArLFHpfg+HaxNJ7FFggwzj7FDq9GmBDRGNCZ06pAK1zJcXCW9Isv5834zI4GvD7gKeqQtk= ; 11, 19 -- Message-ID: {20061107144315.85795.qmail-at-web27901.mail.ukl.yahoo.com} 11, 19 -- Received: from [193.190.208.38] by web27901.mail.ukl.yahoo.com via HTTP; Tue, 07 Nov 2006 14:43:15 GMT 11, 19 -- Date: Tue, 7 Nov 2006 14:43:15 +0000 (GMT) 11, 19 -- From: Samuel Meulenyzer {samuel_meulenyzer-at-yahoo.fr} 11, 19 -- Subject: [Microscopy] SEM (EDX + WDX) - TEM - X-Ray microtomography - Engineer in material characterizations, micro analysis technique 11, 19 -- To: Microscopy-at-microscopy.com 11, 19 -- MIME-Version: 1.0 11, 19 -- Content-Type: text/plain; charset=iso-8859-1 11, 19 -- Content-Transfer-Encoding: 8bit 11, 19 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA7EhGbd025238 ==============================End of - Headers==============================
A Dark field scope produces an image of brightly illuminated objects on a dark background. It was one of the earliest attempts to increase the resolution and detectability of objects.
Abbe's theory of resolution states that for good resolution you need to capture two of three possible rays. This is assuming that each microscopic object acts as a diffraction grating. This isn't too bad of an assumption. The three rays are one direct, undeviated ray from the sample and two primary diffracted rays. (We like to think of these thought experiment and ray diagrams has two dimensional drawing on a sheet of paper). For some subjects the diffracted rays fall outside of the light accepting ability of the objective and these objects can not be resolved. (Bummer!)
One approach is to tilt or move the condense off center (Many of the old medical grade scopes had this ability). This allows the undeviated ray and one diffracted ray to enter the front lens of the objective. This produces a dark background and white illuminated subjects.
A second approach is to use a central stop in the condenser to block the central ray and allow only highly angled rays to illuminated the subject. The diffracted rays from these angled light rays form the image.
You often need a powerful light source or work in a very dark room with dark adapted eyes because you are throwing away a lot of light with a central stop. Special condensers were, and I believe, are still made to efficiently produce darkfield illumination.
Newer illuminating systems (like phase contrast) have replaced a lot of darkfield work. Still some incredible images can be formed with darkfield. One easy short cut is to use a phase contrast scope and select a condenser phase ring larger than the ring in your objective.
Best wishes...........
Frank Karl Degussa Coproration
pavi_micro-at-yahoo. com To: frank.karl-at-degussa.com cc: 11/07/2006 08:12 Subject: [Microscopy] AskAMicroscopist: dark field AM Please respond to pavi_micro
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pavi_micro-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, November 6, 2006 at 23:57:52 ---------------------------------------------------------------------------
Microscopists, While Frank Karl is undoubtedly correct that the spur to invent dark field was based on resolution and tilting beams, I wanted to mention that there is a useful rubric for classifying types of microscopy, and that is based on contrast. Like this:
In each case, the microscope takes advantage of a specific kind of interaction between light and matter to generate contrast (of course there are overlaps, eg scattering contributes to contrast in a brightfield microscope, etc).
(The Nomarski one may be hard to understand, this kind of optics gives rise to a signal whenever the gradient in optical path in a local region is different than what the gradient is in the background).
Hope this helps, Tobias
} } } What an interesting question. } } A Dark field scope produces an image of brightly illuminated objects on a } dark background. It was one of the earliest attempts to increase the } resolution and detectability of objects. } } Abbe's theory of resolution states that for good resolution you need to } capture two of three possible rays. This is assuming that each microscopic } object acts as a diffraction grating. This isn't too bad of an assumption. } The three rays are one direct, undeviated ray from the sample and two } primary diffracted rays. (We like to think of these thought experiment and } ray diagrams has two dimensional drawing on a sheet of paper). For some } subjects the diffracted rays fall outside of the light accepting ability of } the objective and these objects can not be resolved. (Bummer!) } } One approach is to tilt or move the condense off center (Many of the old } medical grade scopes had this ability). This allows the undeviated ray and } one diffracted ray to enter the front lens of the objective. This produces } a dark background and white illuminated subjects. } } A second approach is to use a central stop in the condenser to block the } central ray and allow only highly angled rays to illuminated the subject. } The diffracted rays from these angled light rays form the image. } } You often need a powerful light source or work in a very dark room with } dark adapted eyes because you are throwing away a lot of light with a } central stop. Special condensers were, and I believe, are still made to } efficiently produce darkfield illumination. } } Newer illuminating systems (like phase contrast) have replaced a lot of } darkfield work. Still some incredible images can be formed with darkfield. } One easy short cut is to use a phase contrast scope and select a condenser } phase ring larger than the ring in your objective. } } Best wishes........... } } Frank Karl } Degussa Coproration } } } } } } } pavi_micro-at-yahoo. } com To: } frank.karl-at-degussa.com } } cc: } 11/07/2006 08:12 Subject: } [Microscopy] AskAMicroscopist: dark } field } } AM } Please respond } to } } pavi_micro } } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Did anyone has an experience with LR White polymerization at the temperature range 45-55C? Could you, please, suggest the appropriate embedding/curing protocol and your vendor of choice? My protocol involves the cryofixation, cryo substitution with acetone (aldehydes or osmium fixatives are no-no). The UV polymerization could be difficult to achieve due to the technical limitations; and temperatures exceeding 55C are damaging for some of the structures. The 46-49C range is absolutely preferred. Thank you in advance, Albina
MIKHAYLOVA,ALBINA, PhD Materials Science and Engineering University of Florida PO Box 116400 Gainesville, Florida 32611 Phone: (352) 392-6533 Fax: (352) 392-3771 E-Mail: amich-at-ufl.edu
==============================Original Headers============================== 5, 22 -- From amich-at-ufl.edu Tue Nov 7 10:09:18 2006 5, 22 -- Received: from smtp.ufl.edu (smtp02.osg.ufl.edu [128.227.74.165]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA7G9I88004236 5, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 10:09:18 -0600 5, 22 -- Received: from osgjas02.cns.ufl.edu (osgjas02.cns.ufl.edu [128.227.74.132]) 5, 22 -- by smtp.ufl.edu (8.13.7/8.13.7/2.5.9) with ESMTP id kA7G992M1474576 5, 22 -- for {Microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 11:09:10 -0500 5, 22 -- Message-ID: {1156372807.74371162915749374.JavaMail.osg-at-osgjas02.cns.ufl.edu} 5, 22 -- Date: Tue, 7 Nov 2006 11:09:09 -0500 (EST) 5, 22 -- From: "MIKHAYLOVA,ALBINA" {amich-at-ufl.edu} 5, 22 -- To: Microscopy-at-microscopy.com 5, 22 -- Subject: LR white question 5, 22 -- MIME-Version: 1.0 5, 22 -- Content-Type: text/plain; format=flowed; charset=us-ascii 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- X-Mailer: GatorMail WebMail (http://GatorMail.sf.net/) 5, 22 -- X-Originating-IP: 72.155.89.76 [72.155.89.76] 5, 22 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-3.0rc3 (smtp.ufl.edu [128.227.74.56]); Tue, 07 Nov 2006 11:09:10 -0500 (EST) 5, 22 -- X-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 5, 22 -- X-UFL-Spam-Status: hits=-1.44, required=5, tests=ALL_TRUSTED 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==============================
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America Hi Alwyn,
I wonder if terahertz microwave imaging might work?
Airport security are experimenting with this in the UK - apparently, it can image, in real time, underneath people's clothes to see if hidden weapons, etc, are being carried. Seems a lot more sensitive and controllable than X-rays. Various concerns about security staff having fun watching 'naked' passengers ....
Perhaps some careful tuning would make it possible image at various depths and select different materials to 'see' this writing ...? -- Larry Stoter
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==============================Original Headers============================== 6, 18 -- From larry-at-cymru.freewire.co.uk Tue Nov 7 14:33:07 2006 6, 18 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA7KX646030998 6, 18 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Nov 2006 14:33:06 -0600 6, 18 -- Received: from [217.154.253.197] (th6dc-217-154-253-197.dial.mistral.co.uk [217.154.253.197]) 6, 18 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id kA7KWqDu017101; 6, 18 -- Tue, 7 Nov 2006 20:32:57 GMT 6, 18 -- Mime-Version: 1.0 6, 18 -- Message-Id: {p06210201c1769be24691-at-[217.154.250.120]} 6, 18 -- In-Reply-To: {200611061931.kA6JVuRr028680-at-ns.microscopy.com} 6, 18 -- References: {200611061931.kA6JVuRr028680-at-ns.microscopy.com} 6, 18 -- Date: Tue, 7 Nov 2006 20:28:44 +0000 6, 18 -- To: jae5-at-lehigh.edu, Microscopy-at-MSA.Microscopy.Com 6, 18 -- From: Larry Stoter {larry-at-cymru.freewire.co.uk} 6, 18 -- Subject: Re: [Microscopy] Reading inside a solid object 6, 18 -- Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" 6, 18 -- Content-Transfer-Encoding: 8bit 6, 18 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA7KX646030998 ==============================End of - Headers==============================
My question is: What type of commercially available membrane such as a type of millipore filter could be used as a substrate to grow cells (keratinocytes) and subsequently fix, cryoprotect and ultrathin cryosection the membrane/cells intact? If anyone has done this, I would certainly appreciate some advice.
Robert Underwood Universtiy of Washington Dermatology
==============================Original Headers============================== 4, 21 -- From underwoo-at-u.washington.edu Tue Nov 7 15:45:12 2006 4, 21 -- Received: from mxout7.cac.washington.edu (mxout7.cac.washington.edu [140.142.32.178]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA7LjCYE010615 4, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Nov 2006 15:45:12 -0600 4, 21 -- Received: from hymn10.u.washington.edu (hymn10.u.washington.edu [140.142.13.244]) 4, 21 -- by mxout7.cac.washington.edu (8.13.7+UW06.06/8.13.7+UW06.09) with ESMTP id kA7LjB57010911 4, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 4, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Nov 2006 13:45:11 -0800 4, 21 -- Received: from localhost (localhost [127.0.0.1]) 4, 21 -- by hymn10.u.washington.edu (8.13.7+UW06.06/8.13.7+UW06.09) with ESMTP id kA7LjBZF010953 4, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 7 Nov 2006 13:45:11 -0800 4, 21 -- X-Auth-Received: from [128.208.106.163] by hymn10.u.washington.edu via HTTP; Tue, 07 Nov 2006 13:45:11 PST 4, 21 -- Date: Tue, 7 Nov 2006 13:45:11 -0800 (PST) 4, 21 -- From: Robert A Underwood {underwoo-at-u.washington.edu} 4, 21 -- To: Microscopy List {Microscopy-at-MSA.Microscopy.Com} 4, 21 -- Subject: [Microscopy] cryothin sectioning millipore filter 4, 21 -- Message-ID: {Pine.LNX.4.43.0611071345110.13269-at-hymn10.u.washington.edu} 4, 21 -- MIME-Version: 1.0 4, 21 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 4, 21 -- X-PMX-Version: 5.2.1.279297, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2006.11.7.132933 4, 21 -- 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==============================
A few more Comments on Darkfield - but I like the contrast comparison.
1. Part I - Darkfield history
Darkfield has been around a long time (1800s), including paraboloid and ellipsoid condenser uses.
Classically done with a stop in the condenser or with a reflecting objective.
But can be done with a stop on the objective, at the light source, or another focal plane before the eye.
Modifications of this are:
A) with dispersion staining [promoted by Crossman as well as Brown and McCrone in the 1960s, but actually first described by Dodge in Am. Min. in 1948]
or
B) critical darkfield (you can look up an article by Ted Clarke in Microscopy Today on critical Darkfield which comes from: Havics & Clarke: “Critical Darkfield and Its Application to Asbestos Analysis†presented at Inter/Micro-2002, June 24-27, 2002, Chicago, IL.; there are some nice diagrams of the process of light interaction)
2. Part II - Contrast Issues
Phase
I(x’) proportional to [1 - (2*(phase dif)(x’)) ] I = intensity Thus linear with phase change (RI*thickness) useful for up to 1/2 wavelength Best at {1/10 wavelength
Darkfield
I(x’) proportional to (phase dif)^2 (ingoring secondary terms) Thus not so linear with phase change And useful for outlines or very small particles/differences
Hoffman Modulation Contrast
dI = If*(Phase dif)*(TB - TG)*2cos(theta/w) TB = transmittance in bright area TG = transmittance in gray area W = width of slit Theta = angle from perpedicular to slit Thus non-linear but smoothed
Normaski DIC
I proportional to d(dif phase)/dx (simplified) useful from } 1/10 wavelength Up to 1 wavelength best above 1/2 wavelength
[Source for above: Havics: “Asbestos Fiber Counting by Different Optical Contrast Techniquesâ€, presented at Inter/Micro-2003, July 7-10, 2003, Chicago, IL.; I'm trying to get an article out on this in the next 3 months or so which is why it's at my fingertips]
3. Part III additional Comments
Resolution of Darkfield} Hoffman} Phase } DIC } Brightfield based on my tests using a resolution test slide (HSE/NPL test slide). Fluorescence is too media dependent to make a broad statement but does better than Hoffman using fluorescent spheres and about the same as darkfield.
Resolution does not necessarily mean detectability. Contrast is better for detectability of an object whereas resolution is better for defining the structure once detected.
A Dark background in general increases detectability vs a bright background.
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 Message----- X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu] Sent: Tuesday, November 07, 2006 9:55 AM To: ph2-at-sprynet.com
Microscopists, While Frank Karl is undoubtedly correct that the spur to invent dark field was based on resolution and tilting beams, I wanted to mention that there is a useful rubric for classifying types of microscopy, and that is based on contrast. Like this:
In each case, the microscope takes advantage of a specific kind of interaction between light and matter to generate contrast (of course there are overlaps, eg scattering contributes to contrast in a brightfield microscope, etc).
(The Nomarski one may be hard to understand, this kind of optics gives rise to a signal whenever the gradient in optical path in a local region is different than what the gradient is in the background).
Hope this helps, Tobias
} } } What an interesting question. } } A Dark field scope produces an image of brightly illuminated objects } on a dark background. It was one of the earliest attempts to increase } the resolution and detectability of objects. } } Abbe's theory of resolution states that for good resolution you need to } capture two of three possible rays. This is assuming that each } microscopic object acts as a diffraction grating. This isn't too bad } of an assumption. The three rays are one direct, undeviated ray from } the sample and two primary diffracted rays. (We like to think of these } thought experiment and ray diagrams has two dimensional drawing on a } sheet of paper). For some subjects the diffracted rays fall outside of } the light accepting ability of the objective and these objects can not } be resolved. (Bummer!) } } One approach is to tilt or move the condense off center (Many of the } old medical grade scopes had this ability). This allows the undeviated } ray and one diffracted ray to enter the front lens of the objective. } This produces a dark background and white illuminated subjects. } } A second approach is to use a central stop in the condenser to block } the central ray and allow only highly angled rays to illuminated the } subject. The diffracted rays from these angled light rays form the } image. } } You often need a powerful light source or work in a very dark room with } dark adapted eyes because you are throwing away a lot of light with a } central stop. Special condensers were, and I believe, are still made } to efficiently produce darkfield illumination. } } Newer illuminating systems (like phase contrast) have replaced a lot of } darkfield work. Still some incredible images can be formed with } darkfield. One easy short cut is to use a phase contrast scope and } select a condenser phase ring larger than the ring in your objective. } } Best wishes........... } } Frank Karl } Degussa Coproration } } } } } } } pavi_micro-at-yahoo. } com To: } frank.karl-at-degussa.com } } cc: } 11/07/2006 08:12 Subject: } [Microscopy] AskAMicroscopist: dark } field } } AM } Please respond } to } } pavi_micro } } } } } } } } } } ----------------------------------------------------------------------- } ----- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
Does anyone know anything about this techinique for gradient shaving of thin films to expose the surface for FTIR and other analysis? I was approched about using my microtome for this, but from what I can gather about this machine it is much more specialized, and can measure forces experienced by the knife during cutting. They are only interested in exposing the face for analyis, so I am wondering what kind of knife is used, how the alignment is done, etc. Any information would be appreciated. The citation I have in a couple papers is for a machine by Daipla Wintes, an SAICAS CN-20.
Thanks, Leslie Krupp _____________________ Leslie Krupp IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 3, 34 -- From lkrupp-at-us.ibm.com Tue Nov 7 17:17:07 2006 3, 34 -- Received: from e4.ny.us.ibm.com (e4.ny.us.ibm.com [32.97.182.144]) 3, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA7NH75a004488 3, 34 -- for {microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 17:17:07 -0600 3, 34 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 3, 34 -- by e4.ny.us.ibm.com (8.13.8/8.12.11) with ESMTP id kA7NH7Pn008146 3, 34 -- for {microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 18:17:07 -0500 3, 34 -- Received: from d01av01.pok.ibm.com (d01av01.pok.ibm.com [9.56.224.215]) 3, 34 -- by d01relay04.pok.ibm.com (8.13.6/8.13.6/NCO v8.1.1) with ESMTP id kA7NH6g3150320 3, 34 -- for {microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 18:17:06 -0500 3, 34 -- Received: from d01av01.pok.ibm.com (loopback [127.0.0.1]) 3, 34 -- by d01av01.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id kA7NH6rr030368 3, 34 -- for {microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 18:17:06 -0500 3, 34 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 3, 34 -- by d01av01.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id kA7NH6Wh030365 3, 34 -- for {microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 18:17:06 -0500 3, 34 -- To: microscopy-at-microscopy.com 3, 34 -- MIME-Version: 1.0 3, 34 -- Subject: Surface and Interfacial Cutting Analyis System? 3, 34 -- X-Mailer: Lotus Notes Release 7.0 HF277 June 21, 2006 3, 34 -- Message-ID: {OF62816355.46492074-ON8525721F.007FCF82-8825721F.007FEE8C-at-us.ibm.com} 3, 34 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 3, 34 -- Date: Tue, 7 Nov 2006 18:16:59 -0500 3, 34 -- X-MIMETrack: S/MIME Sign by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF277|June 3, 34 -- 21, 2006) at 11/07/2006 03:17:21 PM, 3, 34 -- Serialize by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF277|June 3, 34 -- 21, 2006) at 11/07/2006 03:17:21 PM, 3, 34 -- Serialize complete at 11/07/2006 03:17:21 PM, 3, 34 -- S/MIME Sign failed at 11/07/2006 03:17:21 PM: The cryptographic key was not 3, 34 -- found, 3, 34 -- Serialize by Router on D01ML604/01/M/IBM(Release 7.0.2HF32 | October 17, 2006) at 3, 34 -- 11/07/2006 18:17:05, 3, 34 -- Serialize complete at 11/07/2006 18:17:05 3, 34 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
It is not easy to find a heat pen in this small isolated country named Austria ;-). The only one I found available is around 600 euros, which I find a LITTLE BIT expensive for such an instrument. Someone told me that a soldering iron would do the trick, and they are available at 10 euros in every general store. Has anyone experienced this electrician tool for EM work? Or perhaps someone has an idea where I could find a "true" heat pen at fair price in our old continent?
Regards,
Stephane
==============================Original Headers============================== 7, 19 -- From nizets2-at-yahoo.com Wed Nov 8 01:24:23 2006 7, 19 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.91.144]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA87ONDc024130 7, 19 -- for {microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 01:24:23 -0600 7, 19 -- Received: (qmail 19776 invoked by uid 60001); 8 Nov 2006 07:24:22 -0000 7, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 19 -- s=s1024; d=yahoo.com; 7, 19 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 19 -- b=CD/Knw0zoRhb0W0iMDibqVsUDkXBXPEl3Qh+txTK/Cr61x4Ncby+KJycSzmyJQDaWSvJLn+8yvk/mjn1mZff9q2+5umgUs4dJKAN8TokIMfik1JsS1gH4s1Nw8fhXB64ufi+djyUMhGvloAR/hit5BIpsAaQON7mVb6JCiQdQPE= ; 7, 19 -- Message-ID: {20061108072422.19774.qmail-at-web37412.mail.mud.yahoo.com} 7, 19 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Tue, 07 Nov 2006 23:24:22 PST 7, 19 -- Date: Tue, 7 Nov 2006 23:24:22 -0800 (PST) 7, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 19 -- Subject: Heat pen 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: text/plain; charset=ascii 7, 19 -- Content-Transfer-Encoding: 8bit 7, 19 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA87ONDc024130 ==============================End of - Headers==============================
Recently I tried successfully the following post-fixation:
3mls h2O 1ml 4% OsO4 0.03grams KFECN (329.26)
I warmly recommend it to any TEM microscopist. It is very easy to prepare and can only improve your image. I really improved my membrane staining, and I think it could improve the staining of glycogen too, but I need more time to appreciate that point.
Regards,
Stephane
----- Original Message ---- X-from: "hyi-at-emory.edu" {hyi-at-emory.edu} To: nizets2-at-yahoo.com Sent: Monday, November 6, 2006 9:49:54 PM
Dear Microscopy Researchers:
I remember someone posted a message last year asking for help with the low contrast problem he was having on his cultured cells. He stated, after receiving a number of replies, that he has been using the same protocol for years and never had problem until then. I have heard a few other people reporting the same thing to me. Personally, I had an episode like that years ago, but then resolved the problem by using freshly made osmium.
Right now I am experiencing the same problem again, and the problem persisted even when I used newly ordered osmium solution (4% aqueous solution). I also used very short dehydration time to minimize potential lipid lose, but no visible improvement in contrast.
I still think this problem has something to do with osmium. But I would like to see if anyone out there has any new insight.
Thank you in advance.
Hong
Emory SOM EM
==============================Original Headers============================== 10, 25 -- From hyi-at-emory.edu Mon Nov 6 14:44:39 2006 10, 25 -- Received: from nemausa.cc.emory.edu (nemausa.cc.emory.edu [170.140.8.220]) 10, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA6KicfL006121 10, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 14:44:39 -0600 10, 25 -- Received: from nemausa (nemausa [170.140.8.220]) 10, 25 -- by nemausa.cc.emory.edu (8.13.8/8.13.8) with ESMTP id kA6Kicf6019792 10, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 15:44:38 -0500 (EST) 10, 25 -- Received: from nemausa (unknown [127.0.0.1]) 10, 25 -- by nemausa (Symantec Mail Security) with ESMTP id 632A824F6 10, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 15:44:38 -0500 (EST) 10, 25 -- X-AuditID: aa8c08dc-000000090000171d-06-454f9eb6747b 10, 25 -- Received: from [170.140.233.153] (dhcp233153.wmb.emory.edu [170.140.233.153]) 10, 25 -- by nemausa (Symantec Mail Security) with ESMTP id 326BF24F0 10, 25 -- for {Microscopy-at-microscopy.com} ; Mon, 6 Nov 2006 15:44:38 -0500 (EST) 10, 25 -- Mime-Version: 1.0 (Apple Message framework v622) 10, 25 -- Message-Id: {d6203d1f421b9c295156fd4808ea80b7-at-emory.edu} 10, 25 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 10, 25 -- To: Microscopy-at-microscopy.com 10, 25 -- From: Hong Yi {hyi-at-emory.edu} 10, 25 -- Subject: (Microscopy) Low contrast 10, 25 -- Date: Mon, 6 Nov 2006 15:44:33 -0500 10, 25 -- X-Mailer: Apple Mail (2.622) 10, 25 -- X-Brightmail-Tracker: AAAAAA== 10, 25 -- Content-Transfer-Encoding: 8bit 10, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA6KicfL006121 ==============================End of - Headers==============================
==============================Original Headers============================== 24, 20 -- From nizets2-at-yahoo.com Wed Nov 8 02:00:41 2006 24, 20 -- Received: from web37406.mail.mud.yahoo.com (web37406.mail.mud.yahoo.com [209.191.91.138]) 24, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA880f9x002864 24, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 02:00:41 -0600 24, 20 -- Received: (qmail 6745 invoked by uid 60001); 8 Nov 2006 08:00:41 -0000 24, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 24, 20 -- s=s1024; d=yahoo.com; 24, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:MIME-Version:Content-Type:Content-Transfer-Encoding; 24, 20 -- b=m+tVKJoxJmZo6qB1S4gez17fcczfstvUoxpjKJVS/Hvuh6ydfK2n6MnFxe0yERGWESlZk0LGRrvCiv2OOtmc5nwoAta7wyT9J58vagBfzwlkq126KmYXy2IjMQ12+4dUpEEnuedt/QGKJzzJZH83U1c8bk0IFGCxuPevLif3908= ; 24, 20 -- Message-ID: {20061108080041.6743.qmail-at-web37406.mail.mud.yahoo.com} 24, 20 -- Received: from [80.122.101.102] by web37406.mail.mud.yahoo.com via HTTP; Wed, 08 Nov 2006 00:00:41 PST 24, 20 -- Date: Wed, 8 Nov 2006 00:00:41 -0800 (PST) 24, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 24, 20 -- Subject: Re: [Microscopy] (Microscopy) Low contrast 24, 20 -- To: hyi-at-emory.edu 24, 20 -- Cc: microscopy-at-microscopy.com 24, 20 -- MIME-Version: 1.0 24, 20 -- Content-Type: text/plain; charset=ascii 24, 20 -- Content-Transfer-Encoding: 8bit 24, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA880f9x002864 ==============================End of - Headers==============================
Hi Tony, I thought about dispersion staining, but...
With darkfield we are attempting to manipulate the sample and illumination to get the best possible combination of resolution and detectability. The central stop (or annular) in the back focal plane of the objective helps increase the detectability of the dispersion of light in the sample and mounting media at the expense of resolution. All of the diffracted, image forming rays from the sample have been captured by the front lens. Manipulation of these image forming rays in the back focal plan can increase detectability, add color, make for powerful striking images, but I suspect no further improvement in resolution is possible. I've looked at a lot of chrysotile (as I sure you have) and I've detected very small amounts, but I have never seen an increase in resolution in dispersion staining darkfield or annular stop as compared to good kohler illumination.
Best wishes.....Frank
PS: good luck with the article!!!
ph2-at-sprynet.com To: frank.karl-at-degussa.com 11/07/2006 03:59 cc: PM Subject: [Microscopy] RE: dark field and beyond Please respond to ph2
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
A few more Comments on Darkfield - but I like the contrast comparison.
1. Part I - Darkfield history
Darkfield has been around a long time (1800s), including paraboloid and ellipsoid condenser uses.
Classically done with a stop in the condenser or with a reflecting objective.
But can be done with a stop on the objective, at the light source, or another focal plane before the eye.
Modifications of this are:
A) with dispersion staining [promoted by Crossman as well as Brown and McCrone in the 1960s, but actually first described by Dodge in Am. Min. in 1948]
or
B) critical darkfield (you can look up an article by Ted Clarke in Microscopy Today on critical Darkfield which comes from: Havics & Clarke: “Critical Darkfield and Its Application to Asbestos Analysis†presented at Inter/Micro-2002, June 24-27, 2002, Chicago, IL.; there are some nice diagrams of the process of light interaction)
2. Part II - Contrast Issues
Phase
I(x’) proportional to [1 - (2*(phase dif)(x’)) ] I = intensity Thus linear with phase change (RI*thickness) useful for up to 1/2 wavelength Best at {1/10 wavelength
Darkfield
I(x’) proportional to (phase dif)^2 (ingoring secondary terms) Thus not so linear with phase change And useful for outlines or very small particles/differences
Hoffman Modulation Contrast
dI = If*(Phase dif)*(TB - TG)*2cos(theta/w) TB = transmittance in bright area TG = transmittance in gray area W = width of slit Theta = angle from perpedicular to slit Thus non-linear but smoothed
Normaski DIC
I proportional to d(dif phase)/dx (simplified) useful from } 1/10 wavelength Up to 1 wavelength best above 1/2 wavelength
[Source for above: Havics: “Asbestos Fiber Counting by Different Optical Contrast Techniquesâ€Â, presented at Inter/Micro-2003, July 7-10, 2003, Chicago, IL.; I'm trying to get an article out on this in the next 3 months or so which is why it's at my fingertips]
3. Part III additional Comments
Resolution of Darkfield} Hoffman} Phase } DIC } Brightfield based on my tests using a resolution test slide (HSE/NPL test slide). Fluorescence is too media dependent to make a broad statement but does better than Hoffman using fluorescent spheres and about the same as darkfield.
Resolution does not necessarily mean detectability. Contrast is better for detectability of an object whereas resolution is better for defining the structure once detected.
A Dark background in general increases detectability vs a bright background.
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 Message----- X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu] Sent: Tuesday, November 07, 2006 9:55 AM To: ph2-at-sprynet.com
Microscopists, While Frank Karl is undoubtedly correct that the spur to invent dark field was based on resolution and tilting beams, I wanted to mention that there is a useful rubric for classifying types of microscopy, and that is based on contrast. Like this:
In each case, the microscope takes advantage of a specific kind of interaction between light and matter to generate contrast (of course there are overlaps, eg scattering contributes to contrast in a brightfield microscope, etc).
(The Nomarski one may be hard to understand, this kind of optics gives rise to a signal whenever the gradient in optical path in a local region is different than what the gradient is in the background).
Hope this helps, Tobias
} } } What an interesting question. } } A Dark field scope produces an image of brightly illuminated objects } on a dark background. It was one of the earliest attempts to increase } the resolution and detectability of objects. } } Abbe's theory of resolution states that for good resolution you need to } capture two of three possible rays. This is assuming that each } microscopic object acts as a diffraction grating. This isn't too bad } of an assumption. The three rays are one direct, undeviated ray from } the sample and two primary diffracted rays. (We like to think of these } thought experiment and ray diagrams has two dimensional drawing on a } sheet of paper). For some subjects the diffracted rays fall outside of } the light accepting ability of the objective and these objects can not } be resolved. (Bummer!) } } One approach is to tilt or move the condense off center (Many of the } old medical grade scopes had this ability). This allows the undeviated } ray and one diffracted ray to enter the front lens of the objective. } This produces a dark background and white illuminated subjects. } } A second approach is to use a central stop in the condenser to block } the central ray and allow only highly angled rays to illuminated the } subject. The diffracted rays from these angled light rays form the } image. } } You often need a powerful light source or work in a very dark room with } dark adapted eyes because you are throwing away a lot of light with a } central stop. Special condensers were, and I believe, are still made } to efficiently produce darkfield illumination. } } Newer illuminating systems (like phase contrast) have replaced a lot of } darkfield work. Still some incredible images can be formed with } darkfield. One easy short cut is to use a phase contrast scope and } select a condenser phase ring larger than the ring in your objective. } } Best wishes........... } } Frank Karl } Degussa Coproration } } } } }
} } pavi_micro-at-yahoo.
} com To: } frank.karl-at-degussa.com } } cc:
} 11/07/2006 08:12 Subject: } [Microscopy] AskAMicroscopist: dark } field } } AM
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} } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (pavi_micro-at-yahoo.com) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on } Monday, November 6, 2006 at 23:57:52 } ----------------------------------------------------------------------- } ---- } } Email: pavi_micro-at-yahoo.com } Name: mrs.pavitra jain } } Organization: Microbiologist } } Education: Graduate College } } Location: Hubli,India } } Question: What is dark field microscope } } ----------------------------------------------------------------------- } ---- } } ==============================Original } Headers============================== } 7, 12 -- From zaluzec-at-ultra5.microscopy.com Tue Nov 7 08:10:19 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 kA7EAJO5003959 } 7, 12 -- for {microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 } 08:10:19 -0600 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) 7, 12 -- } Message-Id: {p06110402c1764435990f-at-[206.69.208.22]} } 7, 12 -- Date: Tue, 7 Nov 2006 08:10:18 -0600 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: pavi_micro-at-yahoo.com (by way of Ask-A-Microscopist) 7, } 12 -- Subject: AskAMicroscopist: dark field 7, 12 -- Content-Type: } text/plain; charset="us-ascii" ==============================End of - } Headers============================== } } } } } ==============================Original } Headers============================== } 32, 18 -- From frank.karl-at-degussa.com Tue Nov 7 08:48:20 2006 } 32, 18 -- Received: from malmailout1.rz.itson.com } (mailout1.degussa.com [193.100.56.173]) } 32, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id kA7EmJDj002211 } 32, 18 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Nov 2006 } 08:48:20 -0600 } 32, 18 -- Received: from mobuscomm01.mail.degussa.com } (uss1026.applications.degussanet.com [10.88.88.98]) } 32, 18 -- by malmailout1.rz.itson.com } (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id kA7EmH0A022231; } 32, 18 -- Tue, 7 Nov 2006 15:48:17 +0100 } 32, 18 -- In-Reply-To: {200611071412.kA7EC5LG007708-at-ns.microscopy.com} } 32, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: dark field } 32, 18 -- To: pavi_micro-at-yahoo.com, microscopy-at-msa.microscopy.com } 32, 18 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 } 32, 18 -- Message-ID: } {OF1AB8A2F0.51F14B6F-ON8625721F.004E2BCB-8625721F.00515177-at-degussa.com} } 32, 18 -- From: frank.karl-at-degussa.com } 32, 18 -- Date: Tue, 7 Nov 2006 08:48:13 -0600 } 32, 18 -- X-MIMETrack: Serialize by Router on } MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at } 32, 18 -- 11/07/2006 08:48:18 AM } 32, 18 -- MIME-Version: 1.0 } 32, 18 -- Content-type: text/plain; charset=US-ASCII } ==============================End of - Headers==============================
(1) Does anyone have information about third-party backscatter electron detectors suitable for scanning Auger microprobes (compatible with ultra high vacuum)?
(2) Does anyone have an opinion about using an electron backscatter diffraction detector as the only backscatter electron detector in a microscope?
Clifford Todd The Dow Chemical Co. CTodd2-at-dow.com
==============================Original Headers============================== 4, 22 -- From CTodd2-at-dow.com Wed Nov 8 08:43:45 2006 4, 22 -- Received: from mail86.messagelabs.com (mail86.messagelabs.com [216.82.244.115]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA8EhjB0008092 4, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 08:43:45 -0600 4, 22 -- X-VirusChecked: Checked 4, 22 -- X-Env-Sender: CTodd2-at-dow.com 4, 22 -- X-Msg-Ref: server-11.tower-86.messagelabs.com!1162997021!15799095!1 4, 22 -- X-StarScan-Version: 5.5.10.7; banners=-,-,- 4, 22 -- X-Originating-IP: [216.99.65.27] 4, 22 -- Received: (qmail 8643 invoked from network); 8 Nov 2006 14:43:41 -0000 4, 22 -- Received: from mail7.dow.com (HELO mante88.nam.dow.com) (216.99.65.27) 4, 22 -- by server-11.tower-86.messagelabs.com with SMTP; 8 Nov 2006 14:43:41 -0000 4, 22 -- Received: by mante88.nam.dow.com with Internet Mail Service (5.5.2658.3) 4, 22 -- id {W32RYJ9T} ; Wed, 8 Nov 2006 09:43:40 -0500 4, 22 -- Message-ID: {0F0E6B7B4F6AE84CBC5E7AF57057FBA8077B20-at-USMDLMDOWX022.dow.com} 4, 22 -- From: "Todd, Clifford (C)" {CTodd2-at-dow.com} 4, 22 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 4, 22 -- Subject: backscatter electron detectors 4, 22 -- Date: Wed, 8 Nov 2006 09:44:00 -0500 4, 22 -- MIME-Version: 1.0 4, 22 -- X-Mailer: Internet Mail Service (5.5.2658.3) 4, 22 -- Content-Type: text/plain ==============================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 mlevin-at-forsyth.org as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mlevin-at-forsyth.org Name: Michael Levin
Organization: Forsyth Institute
Title-Subject: [Filtered] need recommendations for embedding medium
Question: Hello all -
I am a developmental biologisy and need a recommendation, to help me choose the right embedding medium. First and simplest, does anyone have a replacement for JB4 that is less toxic? I need something that is transparent (so that I can orient my embryos during embedding), and that I will cut with a vibratome later to make 20 micron sections. I currently use JB4 to section embryos after wholemount immunohistochemistry or in situ hybridization but I need something that is less toxic. Second, I'm looking for a different medium that meets the following criteria. I need to be able to embed soft tissues (frog and chick embryos) for sectioning via vibratome (10 microns to 60 microns thick). Unlike with the JB4, I want to do immunohistochemistry on these sections after cutting (but do not need electron microscopy). The embedding medium also needs to be
1) non-toxic (so that a fume hood is not needed during embedding) 2) solidifies into a block that can be cut with vibratome, but not so hard that I can't trim it later with a razor blade 3) is transparent, so that I can orient embryos while embedding
Can anyone suggest any kind of embedding mix that matches this description? Please email me at mlevin-at-forsyth.org. Thank you in advance!
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Email: bclarke-at-agarscientific.com Name: Bill Clarke
Organization: Agar Scientific Ltd
Title-Subject: [Filtered] Job opportunity
Question: Technical Support Specialist
Agar Scientific is a leading international supplier of consumables, accessories and specialist equipment for all disciplines of microscopy.
The company has a high level of expertise with wide practical experience in specimen preparation and microscopy techniques.
Based in the UK, the primary role is to provide technical support across our diverse product range to our customers and distributors. The successful applicant will also be working closely with our sales and marketing team to develop new business and product lines. Some UK and international travel may be required.
This is a new appointment and the position offers an ideal opportunity for continued development in a small successful company.
Applicants should be of graduate level (or equivalent) in a science or technology related subject and have experience in electron and optical microscopy techniques. They should have a good 'hands on' approach, be confident in front of customers and have good IT and communication skills. The position would suit a scientist or technician who wishes to build on their existing skills in a commercial environment.
Please apply offline to Dr. Lynne Joyce ljoyce-at-agarscientific.com
Actually I already contacted the german distributor (I am in Autria, not Australia - too bad for me ;-)) and they told me that this material was for 110 volts only. Why the Homo sapiens cannot find a worldwide consensus on the current frequency is out of my capacity of comprehension but it is a fact I have to live with. Apparently I have no other choice than to order a portable heat pen and a kit of rechargeable batteries. Thanks to all for the replies and don't hesitate to add more remarks.
Regards,
Stephane
----- Original Message ---- X-from: "Clarkson, Donna R Contr USAMRD/MCMR-UWB-L" {Donna.Clarkson-at-BROOKS.AF.MIL} To: nizets2-at-yahoo.com Sent: Wednesday, November 8, 2006 3:39:00 PM
Dear colleagues,
It is not easy to find a heat pen in this small isolated country named Austria ;-). The only one I found available is around 600 euros, which I find a LITTLE BIT expensive for such an instrument. Someone told me that a soldering iron would do the trick, and they are available at 10 euros in every general store. Has anyone experienced this electrician tool for EM work? Or perhaps someone has an idea where I could find a "true" heat pen at fair price in our old continent?
Regards,
Stephane
==============================Original Headers============================== 7, 19 -- From nizets2-at-yahoo.com Wed Nov 8 01:24:23 2006 7, 19 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.91.144]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA87ONDc024130 7, 19 -- for {microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 01:24:23 -0600 7, 19 -- Received: (qmail 19776 invoked by uid 60001); 8 Nov 2006 07:24:22 -0000 7, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 19 -- s=s1024; d=yahoo.com; 7, 19 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Con tent-Transfer-Encoding; 7, 19 -- b=CD/Knw0zoRhb0W0iMDibqVsUDkXBXPEl3Qh+txTK/Cr61x4Ncby+KJycSzmyJQDaWSvJLn +8yvk/mjn1mZff9q2+5umgUs4dJKAN8TokIMfik1JsS1gH4s1Nw8fhXB64ufi+djyUMhGvlo AR/hit5BIpsAaQON7mVb6JCiQdQPE= ; 7, 19 -- Message-ID: {20061108072422.19774.qmail-at-web37412.mail.mud.yahoo.com} 7, 19 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Tue, 07 Nov 2006 23:24:22 PST 7, 19 -- Date: Tue, 7 Nov 2006 23:24:22 -0800 (PST) 7, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 19 -- Subject: Heat pen 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: text/plain; charset=ascii 7, 19 -- Content-Transfer-Encoding: 8bit 7, 19 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA87ONDc024130 ==============================End of - Headers==============================
==============================Original Headers============================== 27, 19 -- From nizets2-at-yahoo.com Wed Nov 8 09:03:25 2006 27, 19 -- Received: from web37405.mail.mud.yahoo.com (web37405.mail.mud.yahoo.com [209.191.91.137]) 27, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA8F3OAS028721 27, 19 -- for {microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 09:03:25 -0600 27, 19 -- Received: (qmail 80265 invoked by uid 60001); 8 Nov 2006 15:03:24 -0000 27, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 27, 19 -- s=s1024; d=yahoo.com; 27, 19 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 27, 19 -- b=1t5mfm0CKFqmX5G6AXJVQ/v02pBiIc7EXU+mP6q6NvTygDMA0rOYm3169s56+1ymgAcPSWrbmtvsax5ezwEV1u0gZxFBFDTso0nHo5J3B0Sm0jpSZJLqFIojHvkcWH3PGEzdcNyKU5RFmu9Xlg2abOEp7XpjRKBYFxdFql7DuIo= ; 27, 19 -- Message-ID: {20061108150324.80263.qmail-at-web37405.mail.mud.yahoo.com} 27, 19 -- Received: from [80.122.101.102] by web37405.mail.mud.yahoo.com via HTTP; Wed, 08 Nov 2006 07:03:24 PST 27, 19 -- Date: Wed, 8 Nov 2006 07:03:24 -0800 (PST) 27, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 27, 19 -- Subject: Re: [Microscopy] Heat pen 27, 19 -- To: "Clarkson, Donna R Contr USAMRD/MCMR-UWB-L" {Donna.Clarkson-at-BROOKS.AF.MIL} 27, 19 -- MIME-Version: 1.0 27, 19 -- Content-Type: text/plain; charset=ascii 27, 19 -- Content-Transfer-Encoding: 8bit 27, 19 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA8F3OAS028721 ==============================End of - Headers==============================
EMS in NJ sells a battery powered (wax) heat pen that is fairly cheap and works with thin sections or for wax sealing of boats. Go to http://www.emsdiasum.com. Search for heat pen. Catalog number 72679 $28 USD. Replacement tips: 72679-RT.
One could hook up a 3 volt DC power supply to replace the two AA batteries. I decided not to do that because I got "free" batteries from a storeroom supply and I wanted it to remain portable. It will eat up batteries fast, if used continuously or often.
I had one of those expensive ones with a big filament wire but I used this heat pen because it had a sharp pointed tip. It was small enough for heating thin sections in a diamond knife boat while looking at them through a microtome microscope. EMS shows a photograph of it on their web page.
HTH,
Paul
At 01:25 AM 11/8/06 -0600, you wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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EMS sells many fine products including this one but as an owner of this battery powered pen and a 115 V heat pen from Ted Pella, I find there is no comparison for the purpose of expanding sections. the 115 V pen is significantly better (and significantly more expensive). For wax sealing, the battery pen may be sufficient but I use nail polish so can't directly address that.
At 10:05 AM 11/08/06, 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
The use of a central stop in the objective will decrease resolution.
The use of a stop either in the condenser or at the light source (strong light source) allows better resolution (I've tested this; Dodge also mentions a bit of this in 1948). Ted asked me to publish an article on our paper but after looking at numerous articles over the years, I thought it was rehashing history unless I apply it in more detail to sometime relevant (asbestos or pharmaceutical contaminants). Perhaps I'll finish it off in another year or two.
I believe the improvement is because if the high NA at the critical point, combined with uniformity of background (contrast to phase chnges).
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
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-----Original Message----- X-from: frank.karl-at-degussa.com [mailto:frank.karl-at-degussa.com] Sent: Wednesday, November 08, 2006 6:58 AM To: ph2-at-sprynet.com; microscopy-at-msa.microscopy.com
A few more Comments on Darkfield - but I like the contrast comparison.
1. Part I - Darkfield history
Darkfield has been around a long time (1800s), including paraboloid and ellipsoid condenser uses.
Classically done with a stop in the condenser or with a reflecting objective.
But can be done with a stop on the objective, at the light source, or another focal plane before the eye.
Modifications of this are:
A) with dispersion staining [promoted by Crossman as well as Brown and McCrone in the 1960s, but actually first described by Dodge in Am. Min. in 1948]
or
B) critical darkfield (you can look up an article by Ted Clarke in Microscopy Today on critical Darkfield which comes from: Havics & Clarke: “Critical Darkfield and Its Application to Asbestos Analysis†presented at Inter/Micro-2002, June 24-27, 2002, Chicago, IL.; there are some nice diagrams of the process of light interaction)
2. Part II - Contrast Issues
Phase
I(x’) proportional to [1 - (2*(phase dif)(x’)) ] I = intensity Thus linear with phase change (RI*thickness) useful for up to 1/2 wavelength Best at {1/10 wavelength
Darkfield
I(x’) proportional to (phase dif)^2 (ingoring secondary terms) Thus not so linear with phase change And useful for outlines or very small particles/differences
Hoffman Modulation Contrast
dI = If*(Phase dif)*(TB - TG)*2cos(theta/w) TB = transmittance in bright area TG = transmittance in gray area W = width of slit Theta = angle from perpedicular to slit Thus non-linear but smoothed
Normaski DIC
I proportional to d(dif phase)/dx (simplified) useful from } 1/10 wavelength Up to 1 wavelength best above 1/2 wavelength
[Source for above: Havics: “Asbestos Fiber Counting by Different Optical Contrast Techniquesâ€Â, presented at Inter/Micro-2003, July 7-10, 2003, Chicago, IL.; I'm trying to get an article out on this in the next 3 months or so which is why it's at my fingertips]
3. Part III additional Comments
Resolution of Darkfield} Hoffman} Phase } DIC } Brightfield based on my tests using a resolution test slide (HSE/NPL test slide). Fluorescence is too media dependent to make a broad statement but does better than Hoffman using fluorescent spheres and about the same as darkfield.
Resolution does not necessarily mean detectability. Contrast is better for detectability of an object whereas resolution is better for defining the structure once detected.
A Dark background in general increases detectability vs a bright background.
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
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-----Original Message----- X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu] Sent: Tuesday, November 07, 2006 9:55 AM To: ph2-at-sprynet.com
Microscopists, While Frank Karl is undoubtedly correct that the spur to invent dark field was based on resolution and tilting beams, I wanted to mention that there is a useful rubric for classifying types of microscopy, and that is based on contrast. Like this:
In each case, the microscope takes advantage of a specific kind of interaction between light and matter to generate contrast (of course there are overlaps, eg scattering contributes to contrast in a brightfield microscope, etc).
(The Nomarski one may be hard to understand, this kind of optics gives rise to a signal whenever the gradient in optical path in a local region is different than what the gradient is in the background).
Hope this helps, Tobias
} } } What an interesting question. } } A Dark field scope produces an image of brightly illuminated objects } on a dark background. It was one of the earliest attempts to increase } the resolution and detectability of objects. } } Abbe's theory of resolution states that for good resolution you need to } capture two of three possible rays. This is assuming that each } microscopic object acts as a diffraction grating. This isn't too bad } of an assumption. The three rays are one direct, undeviated ray from } the sample and two primary diffracted rays. (We like to think of these } thought experiment and ray diagrams has two dimensional drawing on a } sheet of paper). For some subjects the diffracted rays fall outside of } the light accepting ability of the objective and these objects can not } be resolved. (Bummer!) } } One approach is to tilt or move the condense off center (Many of the } old medical grade scopes had this ability). This allows the undeviated } ray and one diffracted ray to enter the front lens of the objective. } This produces a dark background and white illuminated subjects. } } A second approach is to use a central stop in the condenser to block } the central ray and allow only highly angled rays to illuminated the } subject. The diffracted rays from these angled light rays form the } image. } } You often need a powerful light source or work in a very dark room with } dark adapted eyes because you are throwing away a lot of light with a } central stop. Special condensers were, and I believe, are still made } to efficiently produce darkfield illumination. } } Newer illuminating systems (like phase contrast) have replaced a lot of } darkfield work. Still some incredible images can be formed with } darkfield. One easy short cut is to use a phase contrast scope and } select a condenser phase ring larger than the ring in your objective. } } Best wishes........... } } Frank Karl } Degussa Coproration } } } } }
} } pavi_micro-at-yahoo.
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} } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (pavi_micro-at-yahoo.com) from } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on } Monday, November 6, 2006 at 23:57:52 } ----------------------------------------------------------------------- } ---- } } Email: pavi_micro-at-yahoo.com } Name: mrs.pavitra jain } } Organization: Microbiologist } } Education: Graduate College } } Location: Hubli,India } } Question: What is dark field microscope } } ----------------------------------------------------------------------- } ---- } } ==============================Original } Headers============================== } 7, 12 -- From zaluzec-at-ultra5.microscopy.com Tue Nov 7 08:10:19 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 kA7EAJO5003959 } 7, 12 -- for {microscopy-at-microscopy.com} ; Tue, 7 Nov 2006 } 08:10:19 -0600 } 7, 12 -- Mime-Version: 1.0 } 7, 12 -- X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) 7, 12 -- } Message-Id: {p06110402c1764435990f-at-[206.69.208.22]} } 7, 12 -- Date: Tue, 7 Nov 2006 08:10:18 -0600 } 7, 12 -- To: microscopy-at-microscopy.com } 7, 12 -- From: pavi_micro-at-yahoo.com (by way of Ask-A-Microscopist) 7, } 12 -- Subject: AskAMicroscopist: dark field 7, 12 -- Content-Type: } text/plain; charset="us-ascii" ==============================End of - } Headers============================== } } } } } ==============================Original } Headers============================== } 32, 18 -- From frank.karl-at-degussa.com Tue Nov 7 08:48:20 2006 32, 18 } -- Received: from malmailout1.rz.itson.com (mailout1.degussa.com } [193.100.56.173]) } 32, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id kA7EmJDj002211 } 32, 18 -- for {microscopy-at-msa.microscopy.com} ; Tue, 7 Nov 2006 } 08:48:20 -0600 } 32, 18 -- Received: from mobuscomm01.mail.degussa.com } (uss1026.applications.degussanet.com [10.88.88.98]) } 32, 18 -- by malmailout1.rz.itson.com } (8.13.4/8.13.4/Debian-3sarge3) with ESMTP id kA7EmH0A022231; } 32, 18 -- Tue, 7 Nov 2006 15:48:17 +0100 } 32, 18 -- In-Reply-To: {200611071412.kA7EC5LG007708-at-ns.microscopy.com} } 32, 18 -- Subject: Re: [Microscopy] AskAMicroscopist: dark field 32, 18 } -- To: pavi_micro-at-yahoo.com, microscopy-at-msa.microscopy.com 32, 18 -- } X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 32, 18 -- } Message-ID: } {OF1AB8A2F0.51F14B6F-ON8625721F.004E2BCB-8625721F.00515177-at-degussa.com} } 32, 18 -- From: frank.karl-at-degussa.com } 32, 18 -- Date: Tue, 7 Nov 2006 08:48:13 -0600 } 32, 18 -- X-MIMETrack: Serialize by Router on } MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 32, 18 -- } 11/07/2006 08:48:18 AM 32, 18 -- MIME-Version: 1.0 } 32, 18 -- Content-type: text/plain; charset=US-ASCII } ==============================End of - Headers==============================
(1) KE and Robinson make scintillator BSD units. The key I think for ultra high vacuum is the use of bellows rather than O-ring seals. Robinson makes this. Not sure about KE. The other option is a solid state BSD which just needs a feed through port for the diode signal wires. But performance is not as good as with scintillator units.
(2) Do you mean EBSD? If so, then the answer is no. EBSD is totally different from BSD. BSD is elemental contrast while EBSD is crystal lattice info. However, using EDS together with EBSD can produce very useful results for multi-phase specimens, if applicable. Note that EBSD does not work on amorphous specimens or areas.
gary g.
At 06:45 AM 11/8/2006, you wrote:
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==============================Original Headers============================== 9, 20 -- From gary-at-gaugler.com Wed Nov 8 10:45:30 2006 9, 20 -- Received: from qsmtp3.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 kA8GjTa1019416 9, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 10:45:29 -0600 9, 20 -- Received: (qmail 13091 invoked from network); 8 Nov 2006 08:45:29 -0800 9, 20 -- Received: by simscan 1.1.0 ppid: 13075, pid: 13088, t: 0.1712s 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 qsmtp3 with SMTP; 8 Nov 2006 08:45:29 -0800 9, 20 -- Message-Id: {7.0.1.0.2.20061108083853.024c7570-at-gaugler.com} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 20 -- Date: Wed, 08 Nov 2006 08:45:28 -0800 9, 20 -- To: CTodd2-at-dow.com 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] backscatter electron detectors 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200611081445.kA8EjlZI011023-at-ns.microscopy.com} 9, 20 -- References: {200611081445.kA8EjlZI011023-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 don't have a suggestion for a less toxic replacement for JB4 but agar may do the trick for your second embedding proposition. You could try embedding in 3% low gel point agar which liquifies fairly quickly in a waterbath at 60'C, stays liquid to about 35'C and is quite solid at RT. If necesary (and if it doesn't compromise your subsequent procedures), you can probably get better support / better sections by immersing in fixative (aldehyde or ethanol) to make the gel more solid.
Hope this helps,
Alastair At 09:06 08/11/2006 -0600, you wrote:
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Alastair McKinnon Histology & EM Facility Manager University of Aberdeen, Institute of Medical Science Foresterhill, Aberdeen, AB25 2ZD tel: +44(0) 1224 552923 (office); +44(0) 1224 555911 (lab) fax: +44(0) 1224 559533; email: a.d.mckinnon-at-abdn.ac.uk www.abdn.ac.uk/ims/h-em
==============================Original Headers============================== 10, 19 -- From a.d.mckinnon-at-abdn.ac.uk Wed Nov 8 11:38:32 2006 10, 19 -- Received: from mailhub3.abdn.ac.uk (mailhub3.abdn.ac.uk [139.133.7.13]) 10, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA8HcVRV031466 10, 19 -- for {Microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 11:38:31 -0600 10, 19 -- Received: from med-0069.ims.abdn.ac.uk ([139.133.159.90] helo=med-0069.abdn.ac.uk) 10, 19 -- by mailhub3.abdn.ac.uk with esmtp (Exim 4.52) 10, 19 -- id 1GhrNV-0003dq-BE; Wed, 08 Nov 2006 17:38:21 +0000 10, 19 -- Message-Id: {5.2.1.1.0.20061108172609.011101d0-at-mailms.abdn.ac.uk} 10, 19 -- X-Sender: pat081-at-mailms.abdn.ac.uk 10, 19 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 10, 19 -- Date: Wed, 08 Nov 2006 17:38:18 +0000 10, 19 -- To: mlevin-at-forsyth.org 10, 19 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 10, 19 -- Subject: Re: [Microscopy] viaWWW: need recommendations for embedding 10, 19 -- medium 10, 19 -- Cc: Microscopy-at-microscopy.com 10, 19 -- In-Reply-To: {200611081506.kA8F6KGX005773-at-ns.microscopy.com} 10, 19 -- Mime-Version: 1.0 10, 19 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Look around for a conversion transformer. These convert between 120 and 240 and vice versa. You would need a unit that would handle the VA or Watts or Amps that the pen requires.
I got one of these at our US Radio Shack store to convert 120 to 240 for an Olympus PM-10AD photo controller. It cost about $24US.
gary g.
At 07:06 AM 11/8/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Wed Nov 8 11:58:58 2006 10, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA8Hwv1i009955 10, 20 -- for {microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 11:58:57 -0600 10, 20 -- Received: (qmail 26489 invoked from network); 8 Nov 2006 09:58:52 -0800 10, 20 -- Received: by simscan 1.1.0 ppid: 26411, pid: 26487, t: 0.2096s 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; 8 Nov 2006 09:58:52 -0800 10, 20 -- Message-Id: {7.0.1.0.2.20061108095545.024e4118-at-gaugler.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 20 -- Date: Wed, 08 Nov 2006 09:58:51 -0800 10, 20 -- To: nizets2-at-yahoo.com 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] Re: Heat pen 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200611081506.kA8F63cG004626-at-ns.microscopy.com} 10, 20 -- References: {200611081506.kA8F63cG004626-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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Title-Subject: [Filtered] Help with settings for Canon EOS D60 on Leica DM LB2
Question: I was wondering if someone would be able to help me sort out the settings for a microscope-camera set up I am trying to use to take images of diatoms.
The set up we have is a Leica DM LB2 High Powered Light Microscope with a Leica C Plan 100x1.25 Oil Magnifying Lens. On top is attachment EF-bayonet Leica HC 1.6x for Canon D-SLR Camera. We then have a Canon EOS D60 camera ontop (its quite an old one as the digital out is not USB!). We have been using the camera on AV as suggested in the bayonet instructions. However, what ever changes we make we cant seam to get the quality of image one would expect.
Does anyone have any ideas of what is needing too be changed?
My BSE system was made by a third party manufacturer, GW Electronics. GW disbanded and was sold to: http://www.kedev.co.uk/
I have not yet purchased anything from these folks, but perhaps they could help...
Woody White BWXT Services
-----Original Message----- X-from: CTodd2-at-dow.com [mailto:CTodd2-at-dow.com] Sent: Wednesday, November 08, 2006 9:44 AM To: White, Woody N.
(1) Does anyone have information about third-party backscatter electron detectors suitable for scanning Auger microprobes (compatible with ultra high vacuum)?
(2) Does anyone have an opinion about using an electron backscatter diffraction detector as the only backscatter electron detector in a microscope?
Clifford Todd The Dow Chemical Co. CTodd2-at-dow.com
==============================Original Headers============================== 4, 22 -- From CTodd2-at-dow.com Wed Nov 8 08:43:45 2006 4, 22 -- Received: from mail86.messagelabs.com (mail86.messagelabs.com [216.82.244.115]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kA8EhjB0008092 4, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 8 Nov 2006 08:43:45 -0600 4, 22 -- X-VirusChecked: Checked 4, 22 -- X-Env-Sender: CTodd2-at-dow.com 4, 22 -- X-Msg-Ref: server-11.tower-86.messagelabs.com!1162997021!15799095!1 4, 22 -- X-StarScan-Version: 5.5.10.7; banners=-,-,- 4, 22 -- X-Originating-IP: [216.99.65.27] 4, 22 -- Received: (qmail 8643 invoked from network); 8 Nov 2006 14:43:41 -0000 4, 22 -- Received: from mail7.dow.com (HELO mante88.nam.dow.com) (216.99.65.27) 4, 22 -- by server-11.tower-86.messagelabs.com with SMTP; 8 Nov 2006 14:43:41 -0000 4, 22 -- Received: by mante88.nam.dow.com with Internet Mail Service (5.5.2658.3) 4, 22 -- id {W32RYJ9T} ; Wed, 8 Nov 2006 09:43:40 -0500 4, 22 -- Message-ID: {0F0E6B7B4F6AE84CBC5E7AF57057FBA8077B20-at-USMDLMDOWX022.dow.com} 4, 22 -- From: "Todd, Clifford (C)" {CTodd2-at-dow.com} 4, 22 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 4, 22 -- Subject: backscatter electron detectors 4, 22 -- Date: Wed, 8 Nov 2006 09:44:00 -0500 4, 22 -- MIME-Version: 1.0 4, 22 -- X-Mailer: Internet Mail Service (5.5.2658.3) 4, 22 -- Content-Type: text/plain ==============================End of - Headers==============================
==============================Original Headers============================== 17, 26 -- From nwwhite-at-bwxt.com Wed Nov 8 14:54:20 2006 17, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 17, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA8KsIW7002395 17, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 8 Nov 2006 14:54:20 -0600 17, 26 -- Received: from ([131.184.13.224]) 17, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.2734684; 17, 26 -- Wed, 08 Nov 2006 15:37:49 -0500 17, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.30]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 17, 26 -- Wed, 8 Nov 2006 14:50:13 -0500 17, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 17, 26 -- Content-class: urn:content-classes:message 17, 26 -- MIME-Version: 1.0 17, 26 -- Content-Type: text/plain; 17, 26 -- charset="us-ascii" 17, 26 -- Subject: RE: [Microscopy] backscatter electron detectors 17, 26 -- Date: Wed, 8 Nov 2006 13:49:32 -0500 17, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D0D1-at-BWXSPO01.BWXS.BWXTECH.NET} 17, 26 -- X-MS-Has-Attach: 17, 26 -- X-MS-TNEF-Correlator: 17, 26 -- Thread-Topic: [Microscopy] backscatter electron detectors 17, 26 -- Thread-Index: AccDSQZuytlkuKLYTp2l93z27hSmnwAHSoIg 17, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 17, 26 -- To: {CTodd2-at-dow.com} , "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 17, 26 -- X-OriginalArrivalTime: 08 Nov 2006 19:50:13.0320 (UTC) FILETIME=[1B095C80:01C7036F] 17, 26 -- Content-Transfer-Encoding: 8bit 17, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA8KsIW7002395 ==============================End of - Headers==============================
Mike A company called London Resin produce a series of acrylic resins including LR White. The resin is suitable for LM sections up to 15-20um or em sections, can be used in a lot of immuno labelling. It is transparent and has low toxicity (you may need to check if that's lower than JB4).
In the UK it is marketed by Agar Scientific Ltd, 66a Cambridge Road, Stansted, Essex, CM24 8DA, UK (Tel 0279 813519). But I'm sure that there will be a US outlet (if that's where you are).
The downside is: 1. it may be more expensive than JB4 2. it needs to be polymerised in the absence of air (oxygen). For e.m we use gelatine capsules filled up and a coverslip or lid on each one to exclude air. 3. it is recommended that thinner sections are cut on glass Ralph knives, but thick ones may not need this (I don't know).
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: mlevin-at-forsyth.org
I know that it could seem very little professional... but I get really good pictures just positioning the digital camera on the ocular of the microscope. You just need a good screen on the camera to control focusing and maybe take more than one picture and choose later the best. I bypassed all problems on attachment and bayonets... I insert measure scale in pictures with ImageJ from a picture of a graduated slide. I am dealing mostly with mammal hair samples, but I tried just for fun with other kind of samples with the same good results
bye!
antonella ______________________________________ Antonella Stravisi Dipartimento di Scienze Animali Università di Udine via S. Mauro, 2 33010 Pagnacco (UD) Italy
==============================Original Headers============================== 8, 26 -- From stravisi-at-uniud.it Thu Nov 9 05:03:24 2006 8, 26 -- Received: from smtp.uniud.it (smtp.uniud.it [158.110.1.131]) 8, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA9B3NbF008773 8, 26 -- for {microscopy-at-microscopy.com} ; Thu, 9 Nov 2006 05:03:23 -0600 8, 26 -- Received: from localhost (nospam.uniud.it [158.110.1.213]) 8, 26 -- by delivery.uniud.it (Postfix) with ESMTP id 5D19647CA0A 8, 26 -- for {microscopy-at-microscopy.com} ; Thu, 9 Nov 2006 12:03:23 +0100 (CET) 8, 26 -- Received: from smtp.uniud.it ([158.110.1.131]) 8, 26 -- by localhost (nospam.uniud.it [158.110.1.213]) (amavisd-new, port 10025) 8, 26 -- with ESMTP id 18113-08 for {microscopy-at-microscopy.com} ; 8, 26 -- Thu, 9 Nov 2006 12:03:23 +0100 (CET) 8, 26 -- Received: from ANTOSTRA.uniud.it (unknown [158.110.130.70]) 8, 26 -- by smtp.uniud.it (Postfix) with ESMTP id BD5D547C6AA 8, 26 -- for {Microscopy-at-Microscopy.Com} ; Thu, 9 Nov 2006 12:03:22 +0100 (CET) 8, 26 -- Message-Id: {6.1.0.6.0.20061109120054.01aabeb0-at-hydrus.cc.uniud.it} 8, 26 -- X-Sender: stravisi-at-hydrus.cc.uniud.it 8, 26 -- X-Mailer: QUALCOMM Windows Eudora Version 6.1.0.6 8, 26 -- Date: Thu, 09 Nov 2006 12:03:12 +0100 8, 26 -- To: Microscopy-at-microscopy.com 8, 26 -- From: Antonella Stravisi {stravisi-at-uniud.it} 8, 26 -- Subject: Re: Help with settings for Canon EOS D60 on Leica DM LB2 8, 26 -- Mime-Version: 1.0 8, 26 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 8, 26 -- X-Virus-Scanned: by amavisd-new at nospam.uniud.it 8, 26 -- Content-Transfer-Encoding: 8bit 8, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA9B3NbF008773 ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (dfine-at-seton.org) from on Thursday, November 9, 2006 at 07:43:43 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both dfine-at-seton.org as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: dfine-at-seton.org Name: Diann Fine
Organization: Brackenridge Hospital
Education: Graduate College
Location: Austin, Texas, USA
Title: Leica EMPT
Question: I would appreciate some information from the EM labs using the Leica EMTP. Is it user-friendly and reliable? Would you purchase it again? Were there any problems or issues with it? Do you run 2 different processing programs together at the same time? Do you run separate procedures for kidney and muscle biopsies? Are different procedures run for each different tissue? Thanks for the replies.
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Email: mkelly-at-fs.fed.us Name: M
Organization: USDA
Title-Subject: [Filtered] LRWhite
Question: I have been using LR White resin for immunolabeling. I have problems with contrast in my samples. Sometimes several sections will be too poor a contrast to get pictures and sometimes part of a section will be just adequate contrast. I understand too long an infiltration time can give you contrast problems. I've cut this down but can only cut it back so far as my material is difficult to get embeded.I would sure appreciate any ideas . Thanks, M
Clifford Todd wrote: =============================================== (1) Does anyone have information about third-party backscatter electron detectors suitable for scanning Auger microprobes (compatible with ultra high vacuum)?
(2) Does anyone have an opinion about using an electron backscatter diffraction detector as the only backscatter electron detector in a microscope?
========================================= To answer the questions:
(1) The Robinson detector is not bakeable, but has been installed and successfully operated in microscopes with chamber vacuums in the vicinity of 10exp-8 torr. I am unaware of other BSE detectors that will operate at lower pressures. The detector is retractable and if you wish to have a bellows retraction mechanism, that is also available. See URL http://www.2spi.com/catalog/instruments/robinson-backscattered-electron-BSE-detector.shtml for further information.
(2) As has been pointed out, EBSD requires a different detector from a BSE detector. But I believe one could operate them simultaneously by the appropriate choice of detectors - a BSE detector above the specimen and an EBSD to one side of the specimen.
Disclaimer: SPI Supplies is the North American distributor for the line of Robinson BSE detectors so we are not exactly a disinterested third party.
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
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 11, 25 -- From cgarber-at-2spi.com Thu Nov 9 08:56:19 2006 11, 25 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 11, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA9EuGI5013154 11, 25 -- for {microscopy-at-msa.microscopy.com} ; Thu, 9 Nov 2006 08:56:17 -0600 11, 25 -- Received: from webmail.idv.net (diskless6.external [209.120.196.50]) 11, 25 -- (authenticated bits=0) 11, 25 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id kA9EuCc1019000 11, 25 -- for {microscopy-at-msa.microscopy.com} ; Thu, 9 Nov 2006 09:56:16 -0500 11, 25 -- X-IDV-FirstRcvd: diskless6.external [209.120.196.50] 11, 25 -- X-IDV-HELO: webmail.idv.net 11, 25 -- X-IDV-Authenticated-User: cgarber 11, 25 -- Received: from 71.230.36.118 (auth. user cgarber-at-mail.2spi.com) 11, 25 -- by webmail.idv.net with HTTP; Thu, 09 Nov 2006 09:56:12 -0500 11, 25 -- To: microscopy-at-msa.microscopy.com 11, 25 -- Subject: BSE and EBSD detectors 11, 25 -- Date: Thu, 09 Nov 2006 09:56:12 -0500 11, 25 -- X-Mailer: IlohaMail/0.9.0 (On: webmail.idv.net) 11, 25 -- Message-ID: {a6Qga3Mz.1163084172.3252680.cgarber-at-mail.2spi.com} 11, 25 -- From: {cgarber-at-2spi.com} 11, 25 -- Bounce-To: {cgarber-at-2spi.com} 11, 25 -- Errors-To: {cgarber-at-2spi.com} 11, 25 -- MIME-Version: 1.0 11, 25 -- Content-Type: text/plain; charset=ISO-8859-1 11, 25 -- Content-Transfer-Encoding: 8bit 11, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kA9EuGI5013154 ==============================End of - Headers==============================
One way to add contrast without affecting antigenicity too much is to add ~0.2-2% tannic acid to fix or freeze-sub solvent, followed by a similar concentration of ferric chloride (rather than OsO4). How strong a concentration you use and for how long varies quite a bit with tissue, and I've only ever processed plant tissues - which often need reasonably long (2 days to a few weeks) infiltrations. I've also played around with the U acetate stain - 2% UA in 70% ethanol for not too long (2-5 min) worked reasonably well on thickish (dark gold) sections of medium grade LRWhite.
good luck,
Rosemary White rosemary.white-at-csiro.au CSIRO Plant Industry ph. 61 (0)2-6246 5475 GPO Box 1600 mob. 61 (0)402 835 973 Canberra, ACT 2601 fax. 61 (0)2-6246 5334 Australia
} From: mkelly-at-fs.fed.us } Reply-To: mkelly-at-fs.fed.us } Date: Thu, 9 Nov 2006 08:07:39 -0600 } To: rosemary.white-at-csiro.au } Subject: [Microscopy] viaWWW: LRWhite } } } } } ---------------------------------------------------------------------------- } 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 mkelly-at-fs.fed.us as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: mkelly-at-fs.fed.us } Name: M } } Organization: USDA } } Title-Subject: [Filtered] LRWhite } } Question: I have been using LR White resin for immunolabeling. I have problems } with contrast in my samples. Sometimes several sections will be too poor a } contrast to get pictures and sometimes part of a section will be just adequate } contrast. I understand too long an infiltration time can give you contrast } problems. I've cut this down but can only cut it back so far as my material is } difficult to get embeded.I would sure appreciate any ideas . } Thanks, M } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 6, 12 -- From zaluzec-at-microscopy.com Thu Nov 9 08:05:58 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 } kA9E5ulj001109 } 6, 12 -- for {microscopy-at-microscopy.com} ; Thu, 9 Nov 2006 08:05:57 -0600 } 6, 12 -- Mime-Version: 1.0 } 6, 12 -- X-Sender: (Unverified) } 6, 12 -- Message-Id: {p06110402c178e634940d-at-[206.69.208.22]} } 6, 12 -- Date: Thu, 9 Nov 2006 08:05:54 -0600 } 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: mkelly-at-fs.fed.us (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: LRWhite } 6, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - Headers============================== }
==============================Original Headers============================== 5, 23 -- From Rosemary.White-at-csiro.au Thu Nov 9 15:12:49 2006 5, 23 -- Received: from act-MTAout1.csiro.au (act-MTAout1.csiro.au [150.229.7.37]) 5, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA9LCltm010426 5, 23 -- for {microscopy-at-microscopy.com} ; Thu, 9 Nov 2006 15:12:48 -0600 5, 23 -- DomainKey-Signature: s=email; d=csiro.au; c=nofws; q=dns; b=q0z5RimATmBXKJ7R1xHLFTOLggixmsPKlUOlrygPDY8OdeS+RoEhw7a+ojjYDF7CDtBZWZOntw44g44f1aObmG8G66+q0jFy1OjIOkgUXZGoAVQxiPVv6Sg8cWhOLKxu; 5, 23 -- Received: from exgw1-mel.nexus.csiro.au ([138.194.3.56]) 5, 23 -- by act-MTAout1.csiro.au with ESMTP; 10 Nov 2006 08:12:46 +1100 5, 23 -- X-IronPort-AV: i="4.09,407,1157292000"; 5, 23 -- d="scan'208"; a="121760345:sNHT2960538686" 5, 23 -- Received: from [152.83.167.45] ([152.83.167.45]) by exgw1-mel.nexus.csiro.au with Microsoft SMTPSVC(5.0.2195.6713); 5, 23 -- Fri, 10 Nov 2006 08:11:12 +1100 5, 23 -- User-Agent: Microsoft-Entourage/10.1.4.030702.0 5, 23 -- Date: Fri, 10 Nov 2006 08:13:46 +1100 5, 23 -- Subject: Re: [Microscopy] viaWWW: LRWhite 5, 23 -- From: Rosemary White {Rosemary.White-at-csiro.au} 5, 23 -- To: {microscopy-at-microscopy.com} 5, 23 -- CC: {mkelly-at-fs.fed.us} 5, 23 -- Message-ID: {C179E53A.1A2C0%Rosemary.White-at-csiro.au} 5, 23 -- In-Reply-To: {200611091407.kA9E7dih005553-at-ns.microscopy.com} 5, 23 -- Mime-version: 1.0 5, 23 -- Content-type: text/plain; charset="US-ASCII" 5, 23 -- Content-transfer-encoding: 7bit 5, 23 -- X-OriginalArrivalTime: 09 Nov 2006 21:11:13.0005 (UTC) FILETIME=[960C31D0:01C70443] ==============================End of - Headers==============================
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Email: mkelly-at-fs.fed.us Name: mkelly
Organization: USDA
Title-Subject: [Filtered] LR White
Question: In my earlier post I left out that I am using a TEM for my imunolabeling work. Thanks again for any help anyone can give.
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Email: martini-at-accurel.com Name: Martin Izquierdo
Organization: Accurel
Title-Subject: [Filtered] DualBeam Operator job
Question: Accurel Systems is currently looking for experienced Dual Beam operators. Accurel is located in Sunnyvale, CA
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Email: swtkeller-at-yahoo.com Name: Sandra Keller
Organization: TEM Lab
Title-Subject: [Filtered] TEM-Looking for a general shallow junction etch recipe for silicon
Question: Hi All: I am looking for a general shallow junction etch/stain recipe and procedure for use with silicon for use in TEM shallow junction delineation. Thanks for being such an important resource. Sandra Keller TEM Lab
How is temperature an issue in scanning Auger microprobes. I'm not familiar with these. If heat is an issue, then I agree that heat is not good for scintillator detectors since the detector tip is basically plastic. However, the current Robinsons are metal coated. But I doubt it is sufficient for very hot environments.
BSD for EBSD basically does not work. The killer is the high tilt angle (70 degrees) of the specimen. What does work is fitting the EBSD camera phosphor screen with a Forward Scatter Detector (FSD). The FSD works just like a BSD but at the high tilt. With properly prepared specimens, FSD images are like 3-D. Very good for estimating ROI and step size.
gary g.
At 06:58 AM 11/9/2006, you wrote:
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Gary Gaugler wrote: ================================================ } Chuck: } } How is temperature an issue in scanning Auger microprobes. } I'm not familiar with these. If heat is an issue, then } I agree that heat is not good for scintillator detectors } since the detector tip is basically plastic. However, the } current Robinsons are metal coated. But I doubt it is } sufficient for very hot environments. } } BSD for EBSD basically does not work. The killer is the } high tilt angle (70 degrees) of the specimen. What does } work is fitting the EBSD camera phosphor screen with a } Forward Scatter Detector (FSD). The FSD works just like } a BSD but at the high tilt. With properly prepared } specimens, FSD images are like 3-D. Very good for estimating } ROI and step size. ================================================== It has been my understanding that temperature can at times be important in an Auger system because the energy of Auger electrons is low and the Auger electrons come from very close to the surface, perhaps the top nm or so. If the surface is contaminated with a gas, or some other surface-sorbed species, the Auger electrons, which are related to the material composition of the surface, would only generate a signal from the contamination layer, not the material underneath that it was intended to study. The conventinal wisdom is that UHV and bake out assure low contamination at the surface to be analyzed.
Auger electrons also have a typical energy of 1 - 2 kV and as such are best excited by lower energy electron beams, usually less than 5 kV, and a BSE detector that responds well to low kV electrons is required for BSE imaging at those voltages.
None of the Robinson designed detectors, including the latest versions is good for heating and bakeout. I am reasonably certain this would be the case for any other BSE detector but of course, I could be wrong about that. For the Robinson BSE detectors, the maximum temperature they will withstand is slightly under 50 C.
I presume Dr. Robinson might be following this conversation and perhaps he can explain this better.
Disclaimer: My firm, SPI Supplies is the North American distributor for the Robinson BSE detector product line so we are not a disinterested third party.
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
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 11, 27 -- From cgarber-at-2spi.com Fri Nov 10 00:43:07 2006 11, 27 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 11, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAA6h6PR032238 11, 27 -- for {microscopy-at-msa.microscopy.com} ; Fri, 10 Nov 2006 00:43:07 -0600 11, 27 -- Received: from yourb27fb1c401 (c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118]) 11, 27 -- (authenticated bits=0) 11, 27 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id kAA6biNP016895; 11, 27 -- Fri, 10 Nov 2006 01:37:51 -0500 11, 27 -- X-IDV-FirstRcvd: c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118] 11, 27 -- X-IDV-HELO: yourb27fb1c401 11, 27 -- X-IDV-Authenticated-User: cgarber 11, 27 -- Message-ID: {00ff01c70492$bb974da0$6401a8c0-at-yourb27fb1c401} 11, 27 -- From: "Garber, Charles A" {cgarber-at-2spi.com} 11, 27 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 11, 27 -- References: {200611092155.kA9LtFHu023763-at-ns.microscopy.com} 11, 27 -- Subject: BSE and EBSD detectors 11, 27 -- Date: Fri, 10 Nov 2006 01:37:39 -0500 11, 27 -- MIME-Version: 1.0 11, 27 -- Content-Type: text/plain; 11, 27 -- format=flowed; 11, 27 -- charset="iso-8859-1"; 11, 27 -- reply-type=original 11, 27 -- Content-Transfer-Encoding: 7bit 11, 27 -- X-Priority: 3 11, 27 -- X-MSMail-Priority: Normal 11, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 11, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
Hi Sandra, As far as I'm aware, the only junction delineation stain that has been used successfully for TEM work is 0.5% HF in Nitric. This has been a well- established technique for looking at junctions in Si for years. A fairly recent example for shallow junctions is given in Yoo et al., Appl. Phys. Letts. 80, 2687 (2002). You do have to take some precautions to get a good result. For example, if your sample is mounted on a Cu grid, you have to protect this from the nitric acid by coating it with lacquer or something similar. I seem to remember Ron Anderson telling me he used to cool the etchant well below room temperature to slow the etch rate down and make it easier to control. HF is a particularly nasty acid, so if you have no experience of it make sure you are properly trained in its use before you start.
-----Original Message----- X-from: swtkeller-at-yahoo.com [mailto:swtkeller-at-yahoo.com] Sent: 09 November 2006 21:23 To: Richard Beanland
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Email: swtkeller-at-yahoo.com Name: Sandra Keller
Organization: TEM Lab
Title-Subject: [Filtered] TEM-Looking for a general shallow junction etch recipe for silicon
Question: Hi All: I am looking for a general shallow junction etch/stain recipe and procedure for use with silicon for use in TEM shallow junction delineation. Thanks for being such an important resource. Sandra Keller TEM Lab
==============================Original Headers============================== 6, 13 -- From zaluzec-at-microscopy.com Thu Nov 9 15:21:50 2006 6, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 6, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kA9LLnLE025324 6, 13 -- for {microscopy-at-microscopy.com} ; Thu, 9 Nov 2006 15:21:50 -0600 6, 13 -- Mime-Version: 1.0 6, 13 -- X-Sender: (Unverified) 6, 13 -- Message-Id: {p06110402c1794c59850a-at-[206.69.208.22]} 6, 13 -- Date: Thu, 9 Nov 2006 15:21:47 -0600 6, 13 -- To: microscopy-at-microscopy.com 6, 13 -- From: swtkeller-at-yahoo.com (by way of MicroscopyListserver) 6, 13 -- Subject: viaWWW: TEM-Looking for a general shallow junction etch recipe 6, 13 -- for silicon 6, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
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==============================Original Headers============================== 19, 32 -- From richard.beanland-at-bookham.com Fri Nov 10 03:19:09 2006 19, 32 -- Received: from mail83.messagelabs.com (mail83.messagelabs.com [195.245.231.83]) 19, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAA9J8i5015574 19, 32 -- for {microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 03:19:09 -0600 19, 32 -- X-VirusChecked: Checked 19, 32 -- X-Env-Sender: richard.beanland-at-bookham.com 19, 32 -- X-Msg-Ref: server-2.tower-83.messagelabs.com!1163150347!36555965!1 19, 32 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 19, 32 -- X-Originating-IP: [213.249.209.179] 19, 32 -- Received: (qmail 6882 invoked from network); 10 Nov 2006 09:19:07 -0000 19, 32 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 19, 32 -- by server-2.tower-83.messagelabs.com with SMTP; 10 Nov 2006 09:19:07 -0000 19, 32 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 19, 32 -- Fri, 10 Nov 2006 09:23:11 +0000 19, 32 -- Content-class: urn:content-classes:message 19, 32 -- MIME-Version: 1.0 19, 32 -- Content-Type: text/plain; 19, 32 -- charset="us-ascii" 19, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 19, 32 -- Subject: RE: [Microscopy] viaWWW: TEM-Looking for a general shallow junction etch recipe 19, 32 -- Date: Fri, 10 Nov 2006 09:22:59 -0000 19, 32 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E243B55-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 19, 32 -- X-MS-Has-Attach: 19, 32 -- X-MS-TNEF-Correlator: 19, 32 -- Thread-Topic: [Microscopy] viaWWW: TEM-Looking for a general shallow junction etch recipe 19, 32 -- Thread-Index: AccERh2Jg21Ab3dyRr+Ta7MxJY42AwAYDYOg 19, 32 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 19, 32 -- To: {swtkeller-at-yahoo.com} 19, 32 -- Cc: {microscopy-at-microscopy.com} 19, 32 -- X-OriginalArrivalTime: 10 Nov 2006 09:23:11.0540 (UTC) FILETIME=[D788CF40:01C704A9] 19, 32 -- Content-Transfer-Encoding: 8bit 19, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAA9J8i5015574 ==============================End of - Headers==============================
Hi folks, I have a failure analysis job looking at a flip-chip Si die that has failed. It's been a (long) while since I did failure analysis looking at silicon chips and printed circuit boards.. What strategies are used to get a solder-bump chip off the board for further investigation? Various ploys using a soldering iron and tweezers come to mind but it would be nice to know how the experts do it, if you're willing to share the knowledge.
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I would like to know the tricks for getting Word to write a number with a bar over it (as is used in crystallographic indices). I believe that this has been the subject of past posts and I have tried to find them in the archive but perhaps I am not using the right search terms. My apologies for bringing it up again.
I have had a solution that has served me well for a decade, but now I have a situation where it does not work. I recall that several solutions were proposed and I would like to try another. Help please.
If you are interested, what I have been doing until now is to create the symbol (eg bar-one) with the equation editor and then making an "autotext" entry to insert it.
Thanks, Alwyn -- ........... 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
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Email: ptomic-at-ciclonsemi.com Name: Peter Tomic
Organization: Ciclon Semiconductor Device Corp
Title-Subject: [Filtered] Failure analysis;flip-chip dies
Question: Richard,
A possible solution to this problem is a hot air stream system. If you know the solder composition, e.g. Pb-Sn or Pb-free, you can adjust the air temperature to accomodate. You're likely to fracture the device if you use a conventional soldering iron.
I have one other suggestion as an analysis tool prior to P.C. board removal. If the substrate is not too heavily doped, you may use NIR [Near Infra-Red] microscopy to image the device from the backside. Of course this is provided no NIR opaque materials are present. I'm assuming the device you are working with is Si, but NIR also works well for GaAs.
Regards,
Peter Tomic Ciclon Semiconductor Device Corp.
Hi folks, I have a failure analysis job looking at a flip-chip Si die that has failed. It's been a (long) while since I did failure analysis looking at silicon chips and printed circuit boards.. What strategies are used to get a solder-bump chip off the board for further investigation? Various ploys using a soldering iron and tweezers come to mind but it would be nice to know how the experts do it, if you're willing to share the knowledge.
Many thanks
Richard
________________________________________ Richard Beanland Materials Analysis Bookham Caswell Towcester Northants NN12 8EQ United Kingdom Tel. +44 1327 356362
Alwyn, One possible solution is to install in Windows a special crystallography fonts (crystallography.ttf) available at: http://x-seed.net/freestuff.html. I hope this helps, Leszek
Leszek Kepinski Division of Nanomaterials Chemistry and Catalysis 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 Message ----- X-from: {jae5-at-lehigh.edu} To: {L.Kepinski-at-int.pan.wroc.pl} Sent: Friday, November 10, 2006 2:50 PM
Alwyn, Word allows you to overwrite two characters in the same position by using one of the field code functions. To make "bar-1" you should:
(1) press ctrl-F9 (this will automatically insert two curly brackets { } in which you can type the field-code instruction; (2) inside the curly brackets type EQ \o(1,¯) - you may need to play around to find the "overbar" symbol. On my system I can find this using Insert:Symbol (it is called the "Macron"). Alternatively you can try alt+0175 (using the number keypad). (3) press alt-F9 to display the text (you may need to do this twice). You can toggle between the field-code and the text using Alt-F9.
I seem to remember reading somewhere that for some language settings (where the decimal separator is defined as the comma?) you may need to use a semi-colon instead of a comma to separate the two characters (i.e. (1;¯)).
----- Original Message ----- X-from: {jae5-at-lehigh.edu} To: {awgodfrey-at-mail.tsinghua.edu.cn} Sent: Friday, November 10, 2006 9:50 PM
Chuck,
I just would like to throw in a small clarification on AES as the posts seem to be a little unclears as far as heat usage.
In order to achieve UHV levels of vacuum, the systems have to be baked in order to get all the surfaces clean enough to get to that vacuum level. If you wanted a BSE detector in one of these, I would imagine it would have to be baked also, I don't see a way around that. However, the ability for the detector to withstand temperature, would be not when it's powered up but rather without any power to it. I don't know if this makes any difference with BSE detectors, but I thought it should be pointed out.
UHV is needed in AES so that the surfaces being analyzed are not contaminated by the gas molecules from the vacuum itself. In UHV, the time to form a monolayer of atoms deposited on the surface is a direct function of the level of vacuum acheived. Since AES is a surface analysis technique, meaning the signal is generated from the top 2 or 3 atomic layers (IIRC), it is imperative to be able to keep the surface clean during analysis.
How to achieve a "clean" surface for AES analysis can be complicated, however, I have not seen heating to be a normally used method except in special cases.
The BSE detctor would not have to be able to withstand heat while operating, only while idle and with no power to it.
dj
On Fri, 10 Nov 2006, cgarber-at-2spi.com wrote:
} It has been my understanding that temperature can at times be important in } an Auger system because the energy of Auger electrons is low and the Auger } electrons come from very close to the surface, perhaps the top nm or so. If } the surface is contaminated with a gas, or some other surface-sorbed } species, the Auger electrons, which are related to the material composition } of the surface, would only generate a signal from the contamination layer, } not the material underneath that it was intended to study. The conventinal } wisdom is that UHV and bake out assure low contamination at the surface to } be analyzed. } } Auger electrons also have a typical energy of 1 - 2 kV and as such are best } excited by lower energy electron beams, usually less than 5 kV, and } a BSE detector that responds well to low kV electrons is required for BSE } imaging at those voltages. } } None of the Robinson designed detectors, including the latest versions is } good for heating and bakeout. I am reasonably certain this would be the } case for any other BSE detector but of course, I could be wrong about that. } For the Robinson BSE detectors, the maximum temperature they will withstand } is slightly under 50 C. } } I presume Dr. Robinson might be following this conversation and perhaps he } can explain this better. } } Disclaimer: My firm, SPI Supplies is the North American distributor for the } Robinson BSE detector product line so we are not a disinterested third } party. } } 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 } } } Look for us! } ############################ } WWW: http://www.2spi.com } ############################ } ================================================== } } } } ==============================Original Headers============================== } 11, 27 -- From cgarber-at-2spi.com Fri Nov 10 00:43:07 2006 } 11, 27 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) } 11, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAA6h6PR032238 } 11, 27 -- for {microscopy-at-msa.microscopy.com} ; Fri, 10 Nov 2006 00:43:07 -0600 } 11, 27 -- Received: from yourb27fb1c401 (c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118]) } 11, 27 -- (authenticated bits=0) } 11, 27 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id kAA6biNP016895; } 11, 27 -- Fri, 10 Nov 2006 01:37:51 -0500 } 11, 27 -- X-IDV-FirstRcvd: c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118] } 11, 27 -- X-IDV-HELO: yourb27fb1c401 } 11, 27 -- X-IDV-Authenticated-User: cgarber } 11, 27 -- Message-ID: {00ff01c70492$bb974da0$6401a8c0-at-yourb27fb1c401} } 11, 27 -- From: "Garber, Charles A" {cgarber-at-2spi.com} } 11, 27 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} } 11, 27 -- References: {200611092155.kA9LtFHu023763-at-ns.microscopy.com} } 11, 27 -- Subject: BSE and EBSD detectors } 11, 27 -- Date: Fri, 10 Nov 2006 01:37:39 -0500 } 11, 27 -- MIME-Version: 1.0 } 11, 27 -- Content-Type: text/plain; } 11, 27 -- format=flowed; } 11, 27 -- charset="iso-8859-1"; } 11, 27 -- reply-type=original } 11, 27 -- Content-Transfer-Encoding: 7bit } 11, 27 -- X-Priority: 3 } 11, 27 -- X-MSMail-Priority: Normal } 11, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 } 11, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 } ==============================End of - Headers============================== }
==============================Original Headers============================== 11, 19 -- From dljones-at-bestweb.net Fri Nov 10 08:33:49 2006 11, 19 -- Received: from vms040pub.verizon.net (vms040pub.verizon.net [206.46.252.40]) 11, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAAEXni9020897 11, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 08:33:49 -0600 11, 19 -- Received: from localhost ([71.249.18.226]) 11, 19 -- by vms040.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr 11, 19 -- 3 2006)) with ESMTPA id {0J8I00K2KR3G1LO4-at-vms040.mailsrvcs.net} for 11, 19 -- Microscopy-at-microscopy.com; Fri, 10 Nov 2006 08:33:21 -0600 (CST) 11, 19 -- Date: Fri, 10 Nov 2006 09:46:24 -0500 (Eastern Standard Time) 11, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 11, 19 -- Subject: Re: [Microscopy] BSE and EBSD detectors 11, 19 -- In-reply-to: {200611100647.kAA6l98J006238-at-ns.microscopy.com} 11, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 11, 19 -- To: cgarber-at-2spi.com 11, 19 -- Cc: Microscopy-at-microscopy.com 11, 19 -- Message-id: {Pine.WNT.4.64.0611100918540.1984-at-H-F1} 11, 19 -- MIME-version: 1.0 11, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 11, 19 -- References: {200611100647.kAA6l98J006238-at-ns.microscopy.com} ==============================End of - Headers==============================
Could someone please explain the difference between phase contrast and dark field microcopy? What optic effects one get with one or the other? I see Frank Karl states in his very nice explanation of dark field sent to the list the following: "One easy short cut is to use a phase contrast scope and select a condenser phase ring larger than the ring in your objective". Can Phase contrast be transformed into darkfield then?
Thank you for any explanation
Antonio M Garcia
==============================Original Headers============================== 4, 22 -- From antoniomiguel.garcia-at-gmail.com Fri Nov 10 08:35:01 2006 4, 22 -- Received: from nf-out-0910.google.com (nf-out-0910.google.com [64.233.182.189]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAAEZ0xv023355 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 08:35:00 -0600 4, 22 -- Received: by nf-out-0910.google.com with SMTP id i2so1259922nfe 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 06:34:59 -0800 (PST) 4, 22 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 22 -- s=beta; d=gmail.com; 4, 22 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 4, 22 -- b=qBFefrWuTv3IWzvNp1ZF8HbTVqo3g4aut/8tcZmpXvdrPU1Ht/+dTIizb+WnYe3ADfS4ao5Zp5cINzc3ZBTsAel0e4exv90YBcoN7NH3dBWM7ZDVMx4ximM+OIwuEUD6MUlgebaXMp/14FNqw/GgQ/mwqRZsoVorvJZJTWc90/8= 4, 22 -- Received: by 10.48.48.13 with SMTP id v13mr5455318nfv.1163169299662; 4, 22 -- Fri, 10 Nov 2006 06:34:59 -0800 (PST) 4, 22 -- Received: by 10.48.245.18 with HTTP; Fri, 10 Nov 2006 06:34:59 -0800 (PST) 4, 22 -- Message-ID: {703e88850611100634hddc2e5bofe27d044f29114e6-at-mail.gmail.com} 4, 22 -- Date: Fri, 10 Nov 2006 15:34:59 +0100 4, 22 -- From: "=?ISO-8859-1?Q?Antonio_Miguel_Garc=EDa?=" {antoniomiguel.garcia-at-gmail.com} 4, 22 -- To: Microscopy-at-microscopy.com 4, 22 -- Subject: AskAMicroscopist:Phase contrast vs Dark Field 4, 22 -- MIME-Version: 1.0 4, 22 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- Content-Disposition: inline ==============================End of - Headers==============================
Antonio, Darkfield allows you to detect objects by virtue of their ability to scatter light. Scattering is weak so you have to "get rid" of the non-scattered, incident light. The way you do this is to have the light incident at an angle that is too wide for the lens to gather. Imagine a hollow cone of light coming up through the condenser that is at an angle too wide for the NA of the objective. This hollow cone of light converges on the specimen and carries on past the objective. If the specimen is empty then looking down the tubes you see darkness. That is where darkfield gets its name. But if you have some scatterers, you will see light from them against the dark background, and the more they scatter the brighter they will be. Darkfield is great for picking up dust and debris, among other things. That's the concept. Now, how do you get a hollow cone of light? Well you can see that the diffculty depends on the NA. The higher the NA of your objective the trickier the condenser design needs to be to achive an angle greater than the objective NA. So if you have a modest to low power objective, then a quick and easy way to get such a hollow cone is to use a phase ring designed for a high NA lens. Phase rings exactly provide hollow cones of light and by using a phase ring designed for a high NA objective (ie wide light cone) with a low power objective, the hollow cone of incident light passes that objective by and you get darkfield. Obviously, this approach won't work if you want to do darkfield with a high NA lens, and for that you need a specially designed darkfield condensor.
Now, phase contrast allows you do detect objects by virtue of their optical thickness, which changes the phase of the light passing through them compared to light that does not pass through them. The tricky bit is to convert that into something your eye can see. This is done also with a hollow cone of light but in this case the cone is gathered by the objective and in the rear focal plane of the lens is placed a ring that lines up with the cone. The ring in the obj rear focal plane changes the phase of this incident light, relative any light that goes elsewhere through the rear focal plane of the objective. That is ultimately how the phase difference between light that goes through the sample (and hence elsewhere in the rear focal plane) and the background (light through the phase ring) is detected.
The optics behind phase contrast are much trickier to grasp intuitively, indeed Zernicke won a Nobel prize for inventing it, whereas darkfield has been played with for ages.
hope this helps, Tobias
} } } Hello } } Could someone please explain the difference between phase contrast and dark } field microcopy? What optic effects one get with one or the other? I see Frank } Karl states in his very nice explanation of dark field sent to the list the } following: "One easy short cut is to use a phase contrast scope and select } a condenser phase ring larger than the ring in your objective". Can } Phase contrast } be transformed into darkfield then? } } Thank you for any explanation } } Antonio M Garcia }
At the risk of creating another bump and run... Phase contrast and dark field are different. Phase contrast converts phase differences between the object of interest and the surrounding material to amplitude difference which is what the human eye is sensitive to. One could think of darkfield as a way of increase detectability.
Using a large phase ring in the condenser is a trick. Just a short cut to get illuminated objects on a dark background.
I refer you to Tobias's excellent explanation.
antoniomiguel.garci a-at-gmail.com To: frank.karl-at-degussa.com cc: 11/10/2006 08:35 AM Subject: [Microscopy] AskAMicroscopist:Phase contrast vs Dark Field Please respond to antoniomiguel.garci a
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hello
Could someone please explain the difference between phase contrast and dark field microcopy? What optic effects one get with one or the other? I see Frank Karl states in his very nice explanation of dark field sent to the list the following: "One easy short cut is to use a phase contrast scope and select a condenser phase ring larger than the ring in your objective". Can Phase contrast be transformed into darkfield then?
Thank you for any explanation
Antonio M Garcia
==============================Original Headers============================== 4, 22 -- From antoniomiguel.garcia-at-gmail.com Fri Nov 10 08:35:01 2006 4, 22 -- Received: from nf-out-0910.google.com (nf-out-0910.google.com [64.233.182.189]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAAEZ0xv023355 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 08:35:00 -0600 4, 22 -- Received: by nf-out-0910.google.com with SMTP id i2so1259922nfe 4, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 06:34:59 -0800 (PST) 4, 22 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 22 -- s=beta; d=gmail.com; 4, 22 -- h=received:message-id:date:from:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition;
We have two LKB 7801A knifebreakers which I have been able to bring back up to full functionality, with the help of a lot of useful replies to a fairly recent posting of mine on this listserver. But I have one last question.
When you take the top off the machine, you can see the four heavy screws which hold the breaking head onto its heavy metal base (I'm not talking guitars here). But there are two other, lighter allen-head screws that appear to be there to adjust the height of the breaking head off the metal base when the four heavier screws are tightened down. Is that actually what they are for? Is this to adjust the amount of pressure necessary to break glass?
Thanks for any hints. This is fix-everything-broken week in our lab....
Cheers, Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 7, 23 -- From TindallR-at-missouri.edu Fri Nov 10 09:03:58 2006 7, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAAF3vZZ029981 7, 23 -- for {microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 09:03:57 -0600 7, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 7, 23 -- Fri, 10 Nov 2006 09:03:57 -0600 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: Knifebreaker question 7, 23 -- Date: Fri, 10 Nov 2006 09:03:56 -0600 7, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68B7C-at-UM-XMAIL08.um.umsystem.edu} 7, 23 -- X-MS-Has-Attach: 7, 23 -- X-MS-TNEF-Correlator: 7, 23 -- Thread-Topic: Knifebreaker question 7, 23 -- Thread-Index: AccE2XKSqTGHReuzTz+BnsM/bXsOFQ== 7, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 7, 23 -- To: {microscopy-at-microscopy.com} 7, 23 -- X-OriginalArrivalTime: 10 Nov 2006 15:03:57.0012 (UTC) FILETIME=[71FC4140:01C704D9] 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAAF3vZZ029981 ==============================End of - Headers==============================
Can anybody recommend a good primary antibody against rhodopsin in mice and/or dog retina? Preferably one that has been tested for EM immunogold labeling? My perusal of the online catalogs hasn't been very helpful so far.
Thanks much.
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 6, 23 -- From TindallR-at-missouri.edu Fri Nov 10 10:13:13 2006 6, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAAGDDup011168 6, 23 -- for {microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 10:13:13 -0600 6, 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); 6, 23 -- Fri, 10 Nov 2006 10:13:12 -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: TEM: Anti-rhodopsin ab 6, 23 -- Date: Fri, 10 Nov 2006 10:13:13 -0600 6, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68B80-at-UM-XMAIL08.um.umsystem.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: TEM: Anti-rhodopsin ab 6, 23 -- Thread-Index: AccE4x73PtzboLgUTfqUI9p9BKoOnw== 6, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 6, 23 -- To: {microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 10 Nov 2006 16:13:12.0731 (UTC) FILETIME=[1EFCA6B0:01C704E3] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAAGDDup011168 ==============================End of - Headers==============================
It would depend a great deal on the type of failure mode, or mechanism, you suspect. I have used a hot air heat gun to melt the solder balls and taped the part off the board. I have carefully cut away most of the circuit board, and then ground away the circuit board directly under the module, which would still be attached by the solder balls. I have also used single edge razor blades, or knife blades. Do you plan on powering up the module (or chip), and testing it, after you get it off the board, or is this strictly a physical failure analysis?
The sample prep must not alter/affect the failure, or the clues that are going help solve the puzzle. Heat can be a problem, as well as the water drip while grinding away the circuit board (may need to be done dry, which is difficult with ceramic substrates).
I hope this helps. If there are more specific questions, I will try to help. My 20+ years in FA has been varied, and quite interesting.
Darrell
richard.beanland-at-bookham.com wrote on 11/10/2006 07:51:52 AM:
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} } Hi folks, } I have a failure analysis job looking at a flip-chip Si die that } has failed. It's been a (long) while since I did failure analysis } looking at silicon chips and printed circuit boards.. What strategies } are used to get a solder-bump chip off the board for further } investigation? Various ploys using a soldering iron and tweezers come } to mind but it would be nice to know how the experts do it, if you're } willing to share the knowledge. } } Many thanks } } Richard } } } } ________________________________________ } Richard Beanland } Materials Analysis } Bookham } 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============================== } 9, 31 -- From richard.beanland-at-bookham.com Fri Nov 10 06:50:26 2006 } 9, 31 -- Received: from mail115.messagelabs.com (mail115. } messagelabs.com [195.245.231.179]) } 9, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP } id kAACoPhP030876 } 9, 31 -- for {microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 06:50:26 -0600 } 9, 31 -- X-VirusChecked: Checked } 9, 31 -- X-Env-Sender: richard.beanland-at-bookham.com } 9, 31 -- X-Msg-Ref: server-9.tower-115.messagelabs.com!1163163024!16808388!1 } 9, 31 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- } 9, 31 -- X-Originating-IP: [213.249.209.179] } 9, 31 -- Received: (qmail 10046 invoked from network); 10 Nov 2006 } 12:50:24 -0000 } 9, 31 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM. } ENTERPRISE.PRI) (213.249.209.179) } 9, 31 -- by server-9.tower-115.messagelabs.com with SMTP; 10 Nov } 2006 12:50:24 -0000 } 9, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0. } 223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft } SMTPSVC(6.0.3790.1830); } 9, 31 -- Fri, 10 Nov 2006 12:54:27 +0000 } 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 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 9, 31 -- Subject: Failure analysis;flip-chip dies } 9, 31 -- Date: Fri, 10 Nov 2006 12:54:30 -0000 } 9, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E243B78-at-cas- } smx-02.BOOKHAM.ENTERPRISE.PRI} } 9, 31 -- X-MS-Has-Attach: } 9, 31 -- X-MS-TNEF-Correlator: } 9, 31 -- Thread-Topic: Failure analysis;flip-chip dies } 9, 31 -- Thread-Index: AccEx10DsHw/ctWsTbCzJf2UytBnIg== } 9, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} } 9, 31 -- To: {microscopy-at-microscopy.com} } 9, 31 -- X-OriginalArrivalTime: 10 Nov 2006 12:54:27.0385 (UTC) } FILETIME=[5AED4290:01C704C7] } 9, 31 -- Content-Transfer-Encoding: 8bit } 9, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns. } microscopy.com id kAACoPhP030876 } ==============================End of - Headers==============================
==============================Original Headers============================== 10, 28 -- From milesd-at-us.ibm.com Fri Nov 10 10:59:05 2006 10, 28 -- Received: from e6.ny.us.ibm.com (e6.ny.us.ibm.com [32.97.182.146]) 10, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAAGx41h022762 10, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Nov 2006 10:59:04 -0600 10, 28 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 10, 28 -- by e6.ny.us.ibm.com (8.13.8/8.12.11) with ESMTP id kAAGxLQ0024998 10, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Nov 2006 11:59:21 -0500 10, 28 -- Received: from d01av03.pok.ibm.com (d01av03.pok.ibm.com [9.56.224.217]) 10, 28 -- by d01relay04.pok.ibm.com (8.13.6/8.13.6/NCO v8.1.1) with ESMTP id kAAGwZwJ073774 10, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Nov 2006 11:58:37 -0500 10, 28 -- Received: from d01av03.pok.ibm.com (loopback [127.0.0.1]) 10, 28 -- by d01av03.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id kAAGwZ6a011359 10, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 10 Nov 2006 11:58:35 -0500 10, 28 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com [9.56.229.50]) 10, 28 -- by d01av03.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id kAAGwY1Y011343; 10, 28 -- Fri, 10 Nov 2006 11:58:34 -0500 10, 28 -- In-Reply-To: {200611101251.kAACpqON032202-at-ns.microscopy.com} 10, 28 -- Subject: Re: [Microscopy] Failure analysis;flip-chip dies 10, 28 -- To: richard.beanland-at-bookham.com 10, 28 -- Cc: Microscopy-at-MSA.Microscopy.Com 10, 28 -- X-Mailer: Lotus Notes Release 7.0 HF277 June 21, 2006 10, 28 -- Message-ID: {OF87F6222F.356C316C-ON85257222.005A9C65-85257222.005D407F-at-us.ibm.com} 10, 28 -- From: Darrell Miles {milesd-at-us.ibm.com} 10, 28 -- Date: Fri, 10 Nov 2006 11:58:33 -0500 10, 28 -- X-MIMETrack: Serialize by Router on D01ML076/01/M/IBM(Release 7.0.1FP1 HF1|June 27, 2006) at 10, 28 -- 11/10/2006 11:58:34 10, 28 -- MIME-Version: 1.0 10, 28 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Sent: Friday, November 10, 2006 7:52 AM To: White, Woody N.
Hi folks, I have a failure analysis job looking at a flip-chip Si die that has failed. It's been a (long) while since I did failure analysis looking at silicon chips and printed circuit boards.. What strategies are used to get a solder-bump chip off the board for further investigation? Various ploys using a soldering iron and tweezers come to mind but it would be nice to know how the experts do it, if you're willing to share the knowledge.
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==============================Original Headers============================== 9, 31 -- From richard.beanland-at-bookham.com Fri Nov 10 06:50:26 2006 9, 31 -- Received: from mail115.messagelabs.com (mail115.messagelabs.com [195.245.231.179]) 9, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAACoPhP030876 9, 31 -- for {microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 06:50:26 -0600 9, 31 -- X-VirusChecked: Checked 9, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 9, 31 -- X-Msg-Ref: server-9.tower-115.messagelabs.com!1163163024!16808388!1 9, 31 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 9, 31 -- X-Originating-IP: [213.249.209.179] 9, 31 -- Received: (qmail 10046 invoked from network); 10 Nov 2006 12:50:24 -0000 9, 31 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 9, 31 -- by server-9.tower-115.messagelabs.com with SMTP; 10 Nov 2006 12:50:24 -0000 9, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 9, 31 -- Fri, 10 Nov 2006 12:54:27 +0000 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 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 31 -- Subject: Failure analysis;flip-chip dies 9, 31 -- Date: Fri, 10 Nov 2006 12:54:30 -0000 9, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E243B78-at-cas-smx-02.BOOKHAM.ENTERPRISE.PR I} 9, 31 -- X-MS-Has-Attach: 9, 31 -- X-MS-TNEF-Correlator: 9, 31 -- Thread-Topic: Failure analysis;flip-chip dies 9, 31 -- Thread-Index: AccEx10DsHw/ctWsTbCzJf2UytBnIg== 9, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 9, 31 -- To: {microscopy-at-microscopy.com} 9, 31 -- X-OriginalArrivalTime: 10 Nov 2006 12:54:27.0385 (UTC) FILETIME=[5AED4290:01C704C7] 9, 31 -- Content-Transfer-Encoding: 8bit 9, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAACoPhP030876 ==============================End of - Headers==============================
==============================Original Headers============================== 20, 27 -- From nwwhite-at-bwxt.com Fri Nov 10 12:10:19 2006 20, 27 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 20, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAAIAIJl002715 20, 27 -- for {microscopy-at-msa.microscopy.com} ; Fri, 10 Nov 2006 12:10:19 -0600 20, 27 -- Received: from ([131.184.13.224]) 20, 27 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.2751614; 20, 27 -- Fri, 10 Nov 2006 13:10:00 -0500 20, 27 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.30]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 20, 27 -- Fri, 10 Nov 2006 12:51:20 -0500 20, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 20, 27 -- Content-class: urn:content-classes:message 20, 27 -- MIME-Version: 1.0 20, 27 -- Content-Type: text/plain; 20, 27 -- charset="us-ascii" 20, 27 -- Subject: RE: [Microscopy] Failure analysis;flip-chip dies 20, 27 -- Date: Fri, 10 Nov 2006 12:48:16 -0500 20, 27 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D0D5-at-BWXSPO01.BWXS.BWXTECH.NET} 20, 27 -- X-MS-Has-Attach: 20, 27 -- X-MS-TNEF-Correlator: 20, 27 -- Thread-Topic: [Microscopy] Failure analysis;flip-chip dies 20, 27 -- Thread-Index: AccEyK3fWatR3WJFSrqmIdB25W9RbQAJ0LJg 20, 27 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 20, 27 -- To: {richard.beanland-at-bookham.com} , 20, 27 -- "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 20, 27 -- X-OriginalArrivalTime: 10 Nov 2006 17:51:20.0762 (UTC) FILETIME=[D486E1A0:01C704F0] 20, 27 -- Content-Transfer-Encoding: 8bit 20, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAAIAIJl002715 ==============================End of - Headers==============================
Richard; A lot depends on the exact method of manufacturing of the flip chip part. Every manufacturer does something a little different and then patents their method. Epoxy under fills are popular for taking thermally induced stresses away from the solder bumps. In that case, no amount of hot anything will get the part off. All analysis must be done from the back side. If this sounds difficult, it is. There are a lot of backside thinning techniques that have been developed to address this difficulty. They consume extensive resources but they do work. However, many are proprietary, and cannot be revealed on this forum.
John Mardinly Intel
This is the opinion of the author and does not necessarily represent the opinion of Intel Corp.
Sent: Friday, November 10, 2006 4:51 AM To: Mardinly, John
Hi folks, I have a failure analysis job looking at a flip-chip Si die that has failed. It's been a (long) while since I did failure analysis looking at silicon chips and printed circuit boards.. What strategies are used to get a solder-bump chip off the board for further investigation? Various ploys using a soldering iron and tweezers come to mind but it would be nice to know how the experts do it, if you're willing to share the knowledge.
======================================================================= 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============================== 9, 31 -- From richard.beanland-at-bookham.com Fri Nov 10 06:50:26 2006 9, 31 -- Received: from mail115.messagelabs.com (mail115.messagelabs.com [195.245.231.179]) 9, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAACoPhP030876 9, 31 -- for {microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 06:50:26 -0600 9, 31 -- X-VirusChecked: Checked 9, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 9, 31 -- X-Msg-Ref: server-9.tower-115.messagelabs.com!1163163024!16808388!1 9, 31 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 9, 31 -- X-Originating-IP: [213.249.209.179] 9, 31 -- Received: (qmail 10046 invoked from network); 10 Nov 2006 12:50:24 -0000 9, 31 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 9, 31 -- by server-9.tower-115.messagelabs.com with SMTP; 10 Nov 2006 12:50:24 -0000 9, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 9, 31 -- Fri, 10 Nov 2006 12:54:27 +0000 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 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 31 -- Subject: Failure analysis;flip-chip dies 9, 31 -- Date: Fri, 10 Nov 2006 12:54:30 -0000 9, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E243B78-at-cas-smx-02.BOOKHAM.ENTERPRISE.PR I} 9, 31 -- X-MS-Has-Attach: 9, 31 -- X-MS-TNEF-Correlator: 9, 31 -- Thread-Topic: Failure analysis;flip-chip dies 9, 31 -- Thread-Index: AccEx10DsHw/ctWsTbCzJf2UytBnIg== 9, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 9, 31 -- To: {microscopy-at-microscopy.com} 9, 31 -- X-OriginalArrivalTime: 10 Nov 2006 12:54:27.0385 (UTC) FILETIME=[5AED4290:01C704C7] 9, 31 -- Content-Transfer-Encoding: 8bit 9, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAACoPhP030876 ==============================End of - Headers==============================
==============================Original Headers============================== 19, 31 -- From john.mardinly-at-intel.com Fri Nov 10 12:24:09 2006 19, 31 -- Received: from mga03.intel.com (mga03.intel.com [143.182.124.21]) 19, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAAIO8So013467 19, 31 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 10 Nov 2006 12:24:09 -0600 19, 31 -- Received: from azsmga001.ch.intel.com ([10.2.17.19]) 19, 31 -- by mga03.intel.com with ESMTP; 10 Nov 2006 10:24:08 -0800 19, 31 -- Received: from scsmsx331.sc.intel.com (HELO scsmsx331.amr.corp.intel.com) ([10.3.90.4]) 19, 31 -- by azsmga001.ch.intel.com with ESMTP; 10 Nov 2006 10:24:07 -0800 19, 31 -- X-ExtLoop1: 1 19, 31 -- X-IronPort-AV: i="4.09,410,1157353200"; 19, 31 -- d="scan'208"; a="144318428:sNHT45184867" 19, 31 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx331.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.211); 19, 31 -- Fri, 10 Nov 2006 10:24:07 -0800 19, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 19, 31 -- Content-class: urn:content-classes:message 19, 31 -- MIME-Version: 1.0 19, 31 -- Content-Type: text/plain; 19, 31 -- charset="us-ascii" 19, 31 -- Subject: RE: [Microscopy] Failure analysis;flip-chip dies 19, 31 -- Date: Fri, 10 Nov 2006 10:24:06 -0800 19, 31 -- Message-ID: {1DF4C4D62339DB4C9C98DF04213995B601BEBD2D-at-scsmsx413.amr.corp.intel.com} 19, 31 -- X-MS-Has-Attach: 19, 31 -- X-MS-TNEF-Correlator: 19, 31 -- Thread-Topic: [Microscopy] Failure analysis;flip-chip dies 19, 31 -- Thread-Index: AccExtPcbIhcvQzoQdyMy+YmQEJAhwALLuuA 19, 31 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 19, 31 -- To: {richard.beanland-at-bookham.com} 19, 31 -- Cc: {Microscopy-at-msa.microscopy.com} 19, 31 -- X-OriginalArrivalTime: 10 Nov 2006 18:24:07.0373 (UTC) FILETIME=[68B7E7D0:01C704F5] 19, 31 -- Content-Transfer-Encoding: 8bit 19, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAAIO8So013467 ==============================End of - Headers==============================
This can be (and usually is) a tricky proposition. It would help knowing a few details about your situation before plunging in to strategies.
First off, why do you need to do failure analysis on this IC? This is not meant to sound flippant. If you are really having to do FA on this IC, depending on its complexity, it can be a huge job and even bigger if you are not set up to do this sort of work on a more routine basis. Even so, different types of ICs tend to need different methods of FA. Based on un-installed identical ICs, what type of metal interconnects are used (Al or Cu), how many layers, ILD type and what method of planarization? Is this flip chip on a supporting substrate or flipped directly onto the main PCB? Is there any conformal coating on the board/IC?
What part number is the IC? This will tell what kind of base package it is--plastic or ceramic and the node size of the IC and if it is Pb-free or not. What kind of PCB is the part on? Is it multi-layer FR4 or ? What is the requirement for retaining the main PCB--i.e., can it be destroyed? Once removed from the main PCB, are you going to re-bump the package and do electrical testing? This would be a first look step to test the IC. The problem with this is that depending on the construction of the whole part, the die and/or package could be destroyed such that the whole thing would no longer work electrically.
gary g.
At 04:53 AM 11/10/2006, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Fri Nov 10 12:50:38 2006 11, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAAIobdZ024534 11, 20 -- for {microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 12:50:38 -0600 11, 20 -- Received: (qmail 6249 invoked from network); 10 Nov 2006 10:50:36 -0800 11, 20 -- Received: by simscan 1.1.0 ppid: 6195, pid: 6244, t: 0.1167s 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 qsmtp3 with SMTP; 10 Nov 2006 10:50:36 -0800 11, 20 -- Message-Id: {7.0.1.0.2.20061110103602.024d4cb8-at-gaugler.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 20 -- Date: Fri, 10 Nov 2006 10:50:31 -0800 11, 20 -- To: richard.beanland-at-bookham.com 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] Failure analysis;flip-chip dies 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200611101253.kAACrPpa001481-at-ns.microscopy.com} 11, 20 -- References: {200611101253.kAACrPpa001481-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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Please see the following paper by Dr. Charles W. Pearce, "Defect Delineation and Junction Staining Techniques for Silicon Using Wet Chemistry", AT&T, Allentown, PA presented at the Electrochemical Society, October 1991. The author is just a few doors down from me right now so I can get you questions answered from the source, if need be.
The paper outlines and provides recipes for Sirtl, Secco, Wright-Jenkins, Yang's Etch, Schimmel, Seiter, Dash, Sato and Memc.
Hi Sandra, As far as I'm aware, the only junction delineation stain that has been used successfully for TEM work is 0.5% HF in Nitric. This has been a well- established technique for looking at junctions in Si for years. A fairly recent example for shallow junctions is given in Yoo et al., Appl. Phys. Letts. 80, 2687 (2002). You do have to take some precautions to get a good result. For example, if your sample is mounted on a Cu grid, you have to protect this from the nitric acid by coating it with lacquer or something similar. I seem to remember Ron Anderson telling me he used to cool the etchant well below room temperature to slow the etch rate down and make it easier to control. HF is a particularly nasty acid, so if you have no experience of it make sure you are properly trained in its use before you start.
I'll add another perspective on their descriptions:
Dark Field:
The cone of light that goes through the sample coming out of the condenser is more like a thin annulus. Think of it as a party hat (thin outer rim coming to a point). It is at a very high angle (outside the field of view). The angle that exits the sample layer is too high to be captured by the objective. Think of a party hat on top of another party hat point-to-point. The brim of the top party hat (that should get captured by the objective) is too big. Thus, when no sample is in the light path, the field of view is dark (dakfield). Small specs or the edges of samples (high change in phase) scatter the light so that it changes angle of the rays at that point and does get captured by the objective. One then sees white edges or spots (where this occurs in the sample) over a darkfield (where there is no scattering).
Phase Contrast:
In the most common use, the cone of light that goes through the sample coming out of the condenser is a broad annulus. Think of it as a party hat made out of very thick cardboard (outer rim coming to a point). It is at a moderate angle (inside the field of view). As the light passes through the specimen there are a) rays that are normally seen and do not touch any of the specimen and b) there are some that go through the specimen. The ones that go through the specimen are retarded (many biological specimens retard by 1/4 of a wavelength of light or within a wavelength range). As they come back through the objective, some encounter a dark (quasi opaque) annulus in the back focal plane that matches the optical size of the annulus of light from the condenser. This is called a phase ring. The phase ring set-up is of the correct thickness to either retard the rays by 1/4 of a wavelngth (positive phase contrast) or to advance them (negative phase contrast). The rays going through the specimen are then doubly retarded - hopefully many by 1/4 plus 1/4 = 1/2 wavelength, while other rays are not. When they recombine, those rays that are 1/2 wavelength in difference (out of phase) cancel each other producing darkness. Those that are in phase combine to be bright. This dark (black) vs bright differences provide contrast - hence phase contrast. The actual retardation (phase shift) is dependent upon thickness and refractive indices.
(For positive phase contrast) because many rays do not get their phase shifted, the background is bright with objects or edges dark. For negative phase contrast, the background is dark and the objects are light.
Let's take a phase annulus for phase contrast setup. Let's presume it was designed for an objective of higher numerical aperature (say NA = 0.85 for a 100X obj). If we use an 40X objective with an NA of 0.65, the only the light scattered from the specimen will reach the objective. This will create a darkfield effect. It will not be the optimum though - only the proper sizing will do that.
Another comment, because phase differences are related to thickness, one can see artificial repetitive lines (series or halos) from where thicknesses create phase shifts of 1/4 wavelength plus 1 wavelength, 1/4 wavelength plus 2 wavelengths, 1/4 wavelength plus 3 wavelengths, 1/4 wavelength plus 4 wavelengths, etc. One will also see some effects from 2nd and 3rd harmonics of the main constructive/destructive inference.
And one more comment, because of the phase shifting aspects enhancing edge effects, one can increase detection of Becke line for refractive index matching of isotropic materials (glasses for instances in forensic applications). [This can also be improved with Nomarski DIC under similar principles to an order mag better than typical brightfield]
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
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-----Original Message----- X-from: frank.karl-at-degussa.com [mailto:frank.karl-at-degussa.com] Sent: Friday, November 10, 2006 9:04 AM To: ph2-at-sprynet.com
At the risk of creating another bump and run... Phase contrast and dark field are different. Phase contrast converts phase differences between the object of interest and the surrounding material to amplitude difference which is what the human eye is sensitive to. One could think of darkfield as a way of increase detectability.
Using a large phase ring in the condenser is a trick. Just a short cut to get illuminated objects on a dark background.
I refer you to Tobias's excellent explanation.
antoniomiguel.garci a-at-gmail.com To: frank.karl-at-degussa.com cc: 11/10/2006 08:35 AM Subject: [Microscopy] AskAMicroscopist:Phase contrast vs Dark Field Please respond to antoniomiguel.garci a
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Hello
Could someone please explain the difference between phase contrast and dark field microcopy? What optic effects one get with one or the other? I see Frank Karl states in his very nice explanation of dark field sent to the list the following: "One easy short cut is to use a phase contrast scope and select a condenser phase ring larger than the ring in your objective". Can Phase contrast be transformed into darkfield then?
Thank you for any explanation
Antonio M Garcia
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Hi Fred, Off line I am sending you the instructions I wrote for a CM12 TEM. Good luck, Judy Murphy microscopyproducts Stockton, CA
fahayes-at-ucdavis.edu wrote:
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==============================Original Headers============================== 6, 19 -- From murphyjudy-at-comcast.net Sun Nov 12 12:38:18 2006 6, 19 -- Received: from rwcrmhc12.comcast.net (rwcrmhc12.comcast.net [216.148.227.152]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kACIcH1o005871 6, 19 -- for {Microscopy-at-microscopy.com} ; Sun, 12 Nov 2006 12:38:18 -0600 6, 19 -- Received: from [192.168.1.105] (c-67-181-119-173.hsd1.ca.comcast.net[67.181.119.173]) 6, 19 -- by comcast.net (rwcrmhc12) with ESMTP 6, 19 -- id {20061112183817m1200ktifee} ; Sun, 12 Nov 2006 18:38:17 +0000 6, 19 -- Message-ID: {45576A19.30208-at-comcast.net} 6, 19 -- Date: Sun, 12 Nov 2006 10:38:17 -0800 6, 19 -- From: Judy Murphy {murphyjudy-at-comcast.net} 6, 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 6, 19 -- X-Accept-Language: en-us, en 6, 19 -- MIME-Version: 1.0 6, 19 -- To: Microscopy {Microscopy-at-microscopy.com} 6, 19 -- Subject: Re: [Microscopy] viaWWW: CM12 alignment procedures 6, 19 -- References: {200611121817.kACIHuYu031341-at-ns.microscopy.com} 6, 19 -- In-Reply-To: {200611121817.kACIHuYu031341-at-ns.microscopy.com} 6, 19 -- Content-Type: text/plain; charset=us-ascii; format=flowed 6, 19 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
-- 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============================== 11, 26 -- From r.sims-at-auckland.ac.nz Sun Nov 12 19:47:11 2006 11, 26 -- Received: from groucho.itss.auckland.ac.nz (mailhost.auckland.ac.nz [130.216.190.11]) 11, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAD1lAV5008832 11, 26 -- for {microscopy-at-microscopy.com} ; Sun, 12 Nov 2006 19:47:11 -0600 11, 26 -- Received: from localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) 11, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id 613C33604E 11, 26 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:47:09 +1300 (NZDT) 11, 26 -- Received: from groucho.itss.auckland.ac.nz ([127.0.0.1]) 11, 26 -- by localhost (smtpa.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 11, 26 -- with ESMTP id 10045-13 for {microscopy-at-microscopy.com} ; 11, 26 -- Mon, 13 Nov 2006 14:47:09 +1300 (NZDT) 11, 26 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 11, 26 -- by groucho.itss.auckland.ac.nz (Postfix) with ESMTP id E3ADC3594A 11, 26 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:47:08 +1300 (NZDT) 11, 26 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 11, 26 -- To: microscopy-at-microscopy.com 11, 26 -- Date: Mon, 13 Nov 2006 14:52:03 +1300 11, 26 -- MIME-Version: 1.0 11, 26 -- Subject: Microscopy Conference in New Zealand 11, 26 -- Message-ID: {45588693.7756.1466BC8-at-localhost} 11, 26 -- Priority: normal 11, 26 -- X-mailer: Pegasus Mail for Windows (4.21c) 11, 26 -- Content-type: text/plain; charset=US-ASCII 11, 26 -- Content-transfer-encoding: 7BIT 11, 26 -- Content-description: Mail message body 11, 26 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
Just to give some clarification on this - the chip in question is a Si ASIC, I guess, mounted on a multilayer board which drives a laser, which sends light down an optical fibre; the board slots into a telecoms module. In fact you can see the board at http://www.bookham.com/common/trans_tunable.cfm, or http://www.bookham.com/documents/datasheets_trans/TL5000.pdf. No idea at the moment what the chip does, whether it's bipolar or CMOS, Al or Cu metallisation etc. etc.. There are 8 lead-free solder bumps and there doesn't appear to be any epoxy underfill; the board is expendable. Usually I work on the optoelectronic components produced at the fab here, more recently the miniature optical bench that surrounds it inside a 'gold box', but as we move more into systems rather than just components, things are getting more varied. The failure analysis request at the moment is just "we know this chip has stopped working, what's wrong with it?". I don't think we have the kit here to test the chip's functionality off the board, so if there's nothing obvious like a cracked solder bump or an obvious electrical overstress event I guess it will get pushed back to the PCB supplier.
Some very useful suggestions which I like the sound of: Infra-red through the back of the chip, thanks to Peter Tomic, since this is non-destructive and simple for me to do. Also fuming nitric/sulphuric to get rid of the PCB, thanks Valery Ray. I've already done real-time radiography to look for cracks in the solder and SEM looking around the edges. And perhaps even more important than any technique, knowing some of the right questions to ask about the chip which will make it a lot easier to get a meaningful result.
Thanks to all for a great response to my question,
-----Original Message----- X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com] Sent: 10 November 2006 18:52 To: Richard Beanland
This can be (and usually is) a tricky proposition. It would help knowing a few details about your situation before plunging in to strategies.
First off, why do you need to do failure analysis on this IC? This is not meant to sound flippant. If you are really having to do FA on this IC, depending on its complexity, it can be a huge job and even bigger if you are not set up to do this sort of work on a more routine basis. Even so, different types of ICs tend to need different methods of FA. Based on un-installed identical ICs, what type of metal interconnects are used (Al or Cu), how many layers, ILD type and what method of planarization? Is this flip chip on a supporting substrate or flipped directly onto the main PCB? Is there any conformal coating on the board/IC?
What part number is the IC? This will tell what kind of base package it is--plastic or ceramic and the node size of the IC and if it is Pb-free or not. What kind of PCB is the part on? Is it multi-layer FR4 or ? What is the requirement for retaining the main PCB--i.e., can it be destroyed? Once removed from the main PCB, are you going to re-bump the package and do electrical testing? This would be a first look step to test the IC. The problem with this is that depending on the construction of the whole part, the die and/or package could be destroyed such that the whole thing would no longer work electrically.
gary g.
At 04:53 AM 11/10/2006, you wrote:
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==============================Original Headers============================== 11, 20 -- From gary-at-gaugler.com Fri Nov 10 12:50:38 2006 11, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAAIobdZ024534 11, 20 -- for {microscopy-at-microscopy.com} ; Fri, 10 Nov 2006 12:50:38 -0600 11, 20 -- Received: (qmail 6249 invoked from network); 10 Nov 2006 10:50:36 -0800 11, 20 -- Received: by simscan 1.1.0 ppid: 6195, pid: 6244, t: 0.1167s 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 qsmtp3 with SMTP; 10 Nov 2006 10:50:36 -0800 11, 20 -- Message-Id: {7.0.1.0.2.20061110103602.024d4cb8-at-gaugler.com} 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 20 -- Date: Fri, 10 Nov 2006 10:50:31 -0800 11, 20 -- To: richard.beanland-at-bookham.com 11, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 20 -- Subject: Re: [Microscopy] Failure analysis;flip-chip dies 11, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 20 -- In-Reply-To: {200611101253.kAACrPpa001481-at-ns.microscopy.com} 11, 20 -- References: {200611101253.kAACrPpa001481-at-ns.microscopy.com} 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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==============================Original Headers============================== 27, 31 -- From richard.beanland-at-bookham.com Mon Nov 13 04:42:29 2006 27, 31 -- Received: from mail80.messagelabs.com (mail80.messagelabs.com [195.245.230.163]) 27, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kADAgSh9030669 27, 31 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 04:42:29 -0600 27, 31 -- X-VirusChecked: Checked 27, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 27, 31 -- X-Msg-Ref: server-10.tower-80.messagelabs.com!1163414545!49942741!1 27, 31 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 27, 31 -- X-Originating-IP: [213.249.209.179] 27, 31 -- Received: (qmail 9317 invoked from network); 13 Nov 2006 10:42:25 -0000 27, 31 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 27, 31 -- by server-10.tower-80.messagelabs.com with SMTP; 13 Nov 2006 10:42:25 -0000 27, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 27, 31 -- Mon, 13 Nov 2006 10:46:33 +0000 27, 31 -- Content-class: urn:content-classes:message 27, 31 -- MIME-Version: 1.0 27, 31 -- Content-Type: text/plain; 27, 31 -- charset="us-ascii" 27, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 27, 31 -- Subject: RE: [Microscopy] Failure analysis;flip-chip dies 27, 31 -- Date: Mon, 13 Nov 2006 10:46:34 -0000 27, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E243BCC-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 27, 31 -- X-MS-Has-Attach: 27, 31 -- X-MS-TNEF-Correlator: 27, 31 -- Thread-Topic: [Microscopy] Failure analysis;flip-chip dies 27, 31 -- Thread-Index: AccE+dRd2iIwlm4JQYiPjr78y7W6JwCEm24w 27, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 27, 31 -- To: {microscopy-at-microscopy.com} 27, 31 -- X-OriginalArrivalTime: 13 Nov 2006 10:46:33.0171 (UTC) FILETIME=[FBFA6230:01C70710] 27, 31 -- Content-Transfer-Encoding: 8bit 27, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kADAgSh9030669 ==============================End of - Headers==============================
I manage a multi-user facility and are looking into on-line scheduling programs. I'd appreciate your input as to what's out there and your experiences.
Thanks in advance,
Kathy Troughton Senior Research Associate University of Tennessee, Memphis Memphis, TN 901-448-5976 ktrou-at-nb.utmem.edu
==============================Original Headers============================== 5, 26 -- From ktrou-at-nb.utmem.edu Mon Nov 13 08:32:43 2006 5, 26 -- Received: from mail2.utmem.edu (mail2.utmem.edu [132.192.6.164]) 5, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADEWhlE014007 5, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 08:32:43 -0600 5, 26 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 5, 26 -- by mail2.utmem.edu (SMTP Daemon) with ESMTP id 05DD734080 5, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 08:32:43 -0600 (CST) 5, 26 -- Received: from mail2.utmem.edu ([127.0.0.1]) 5, 26 -- by localhost (mail2.utmem.edu [127.0.0.1]) (amavisd-new, port 10025) 5, 26 -- with ESMTP id 21160-12 for {Microscopy-at-Microscopy.Com} ; 5, 26 -- Mon, 13 Nov 2006 08:32:42 -0600 (CST) 5, 26 -- Received: from nb.utmem.edu (nb.utmem.edu [132.192.43.15]) 5, 26 -- by mail2.utmem.edu (SMTP Daemon) with SMTP id 267B3340D5 5, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 08:32:42 -0600 (CST) 5, 26 -- Received: from [132.192.47.92] (b11701.utmem.edu) by nb.utmem.edu (5.0/SMI-SVR4) 5, 26 -- id AA20804; Mon, 13 Nov 06 07:24:07 CST 5, 26 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 26 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 26 -- Message-Id: {E59D7F8E-054F-4801-9925-427F07728CBF-at-nb.utmem.edu} 5, 26 -- Content-Transfer-Encoding: 7bit 5, 26 -- From: Kathy Troughton {ktrou-at-nb.utmem.edu} 5, 26 -- Subject: Scheduling programs 5, 26 -- Date: Mon, 13 Nov 2006 08:32:38 -0600 5, 26 -- To: Microscopy-at-Microscopy.Com 5, 26 -- X-Mailer: Apple Mail (2.752.2) 5, 26 -- X-Virus-Scanned: by amavisd-new at utmem.edu ==============================End of - Headers==============================
I have designed a dedicated software for scientific multi-user facilities (Facility Online Manager). The detailed features of this software package is discussed in this month's issue of Microscopy Today (November 2006). This software has been used at Northwestern for 3+ years and has been installed in a couple of other institutions.
Take a look here: http://www.fom.northwestern.edu/
Please drop me email if you need demo accounts to the server.
Shuyou _____________________________ Shuyou Li, Ph.D. NUANCE Center Northwestern University 2220 Campus Drive, 1161 Cook Hall Evanston, IL 60208-3108, USA Ph: (847) 491-6723 Fax: (847) 467-6573 Email: syli-at-northwestern.edu http://www.nuance.northwestern.edu/
-----Original Message----- X-from: ktrou-at-nb.utmem.edu [mailto:ktrou-at-nb.utmem.edu] Sent: Monday, November 13, 2006 8:38 AM To: syli-at-northwestern.edu
Hello everyone,
I manage a multi-user facility and are looking into on-line scheduling programs. I'd appreciate your input as to what's out there and your experiences.
Thanks in advance,
Kathy Troughton Senior Research Associate University of Tennessee, Memphis Memphis, TN 901-448-5976 ktrou-at-nb.utmem.edu
==============================Original Headers============================== 5, 26 -- From ktrou-at-nb.utmem.edu Mon Nov 13 08:32:43 2006 5, 26 -- Received: from mail2.utmem.edu (mail2.utmem.edu [132.192.6.164]) 5, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADEWhlE014007 5, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 08:32:43 -0600 5, 26 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 5, 26 -- by mail2.utmem.edu (SMTP Daemon) with ESMTP id 05DD734080 5, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 08:32:43 -0600 (CST) 5, 26 -- Received: from mail2.utmem.edu ([127.0.0.1]) 5, 26 -- by localhost (mail2.utmem.edu [127.0.0.1]) (amavisd-new, port 10025) 5, 26 -- with ESMTP id 21160-12 for {Microscopy-at-Microscopy.Com} ; 5, 26 -- Mon, 13 Nov 2006 08:32:42 -0600 (CST) 5, 26 -- Received: from nb.utmem.edu (nb.utmem.edu [132.192.43.15]) 5, 26 -- by mail2.utmem.edu (SMTP Daemon) with SMTP id 267B3340D5 5, 26 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 08:32:42 -0600 (CST) 5, 26 -- Received: from [132.192.47.92] (b11701.utmem.edu) by nb.utmem.edu (5.0/SMI-SVR4) 5, 26 -- id AA20804; Mon, 13 Nov 06 07:24:07 CST 5, 26 -- Mime-Version: 1.0 (Apple Message framework v752.2) 5, 26 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 5, 26 -- Message-Id: {E59D7F8E-054F-4801-9925-427F07728CBF-at-nb.utmem.edu} 5, 26 -- Content-Transfer-Encoding: 7bit 5, 26 -- From: Kathy Troughton {ktrou-at-nb.utmem.edu} 5, 26 -- Subject: Scheduling programs 5, 26 -- Date: Mon, 13 Nov 2006 08:32:38 -0600 5, 26 -- To: Microscopy-at-Microscopy.Com 5, 26 -- X-Mailer: Apple Mail (2.752.2) 5, 26 -- X-Virus-Scanned: by amavisd-new at utmem.edu ==============================End of - Headers==============================
==============================Original Headers============================== 18, 20 -- From syli-at-northwestern.edu Mon Nov 13 12:25:01 2006 18, 20 -- Received: from merle.it.northwestern.edu (merle.it.northwestern.edu [129.105.16.57]) 18, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADIP1JV007199 18, 20 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 12:25:01 -0600 18, 20 -- Received: from prep (orion.ms.northwestern.edu [129.105.37.66]) 18, 20 -- by merle.it.northwestern.edu (Postfix) with ESMTP id DCEFF2F05F; 18, 20 -- Mon, 13 Nov 2006 12:25:00 -0600 (CST) 18, 20 -- From: "Shuyou Li" {syli-at-northwestern.edu} 18, 20 -- To: {ktrou-at-nb.utmem.edu} , {Microscopy-at-Microscopy.Com} 18, 20 -- Subject: RE: [Microscopy] Scheduling programs 18, 20 -- Date: Mon, 13 Nov 2006 12:24:57 -0600 18, 20 -- Message-ID: {002a01c70751$05bc4b40$42256981-at-prep} 18, 20 -- MIME-Version: 1.0 18, 20 -- Content-Type: text/plain; 18, 20 -- charset="us-ascii" 18, 20 -- Content-Transfer-Encoding: 7bit 18, 20 -- X-Mailer: Microsoft Office Outlook 11 18, 20 -- Thread-Index: AccHMY1f+ngMqTFFSuywWWiDU87xXgAHcHTw 18, 20 -- In-Reply-To: {200611131438.kADEc20u021327-at-ns.microscopy.com} 18, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
I have a problem with PIPS (Gatan) in our lab. The samples (including metal and semiconductors) are not milled at all (like 10 hours at voltage 4.5). We just cleaned the penning gauge and two guns. The vacuum and current looks common. I wonder someone can give me suggestion what is the problem.
Thanks.
Jiaming
==============================Original Headers============================== 5, 16 -- From zhangj16-at-msu.edu Mon Nov 13 12:29:40 2006 5, 16 -- Received: from egr.msu.edu (jeeves.egr.msu.edu [35.9.37.127]) 5, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADITeG3013018 5, 16 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 12:29:40 -0600 5, 16 -- Received: from [127.0.0.1] (muran.dhcp.egr.msu.edu [35.9.138.199]) 5, 16 -- by egr.msu.edu (8.13.7/8.13.4) with ESMTP id kADITdGZ007932 5, 16 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 13:29:41 -0500 (EST) 5, 16 -- Message-ID: {4558B992.6030803-at-msu.edu} 5, 16 -- Date: Mon, 13 Nov 2006 13:29:38 -0500 5, 16 -- From: Jiaming Zhang {zhangj16-at-msu.edu} 5, 16 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) 5, 16 -- MIME-Version: 1.0 5, 16 -- To: Microscopy-at-Microscopy.Com 5, 16 -- Subject: One problem with Ion milling 5, 16 -- Content-Type: text/plain; charset=GB2312 5, 16 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I need to prepare a self supporting cross section of a multilayer film on a 1" silicon wafer. Problem is the material cannot be heated above room temperature. So as far as I know, I need to use something other than G-1 to prepare the sandwich and something other than crystalbond to do the polishing/dimpling. My first thought is M-bond 610 for the sandwich, I've heard this will cure overnight at RT? And superglue for the polishing, although this will require soaking off between switching sides. M-bond 610 is not soluble in acetone, right?
Any other ideas? Commercial responses are welcome.
The sample will be ion-milled at LN2 temperatures after the polishing/dimpling steps.
Thanks, Leslie _____________________ Leslie Krupp IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 5, 27 -- From lkrupp-at-us.ibm.com Mon Nov 13 13:37:14 2006 5, 27 -- Received: from e6.ny.us.ibm.com (e6.ny.us.ibm.com [32.97.182.146]) 5, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADJbEPS029873 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 13:37:14 -0600 5, 27 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 5, 27 -- by e6.ny.us.ibm.com (8.13.8/8.12.11) with ESMTP id kADJbXNw029806 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:33 -0500 5, 27 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) 5, 27 -- by d01relay02.pok.ibm.com (8.13.6/8.13.6/NCO v8.1.1) with ESMTP id kADJbDu5181762 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) 5, 27 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id kADJbD9N006725 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 5, 27 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id kADJbD4q006713 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- To: microscopy-at-microscopy.com 5, 27 -- MIME-Version: 1.0 5, 27 -- Subject: TEM sample prep: no heat adhesives required 5, 27 -- X-Mailer: Lotus Notes Release 7.0 HF277 June 21, 2006 5, 27 -- Message-ID: {OF98894D7C.38D025D7-ON85257225.006B12FC-88257225.006BC206-at-us.ibm.com} 5, 27 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 5, 27 -- Date: Mon, 13 Nov 2006 11:36:31 -0800 5, 27 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.2HF32 | October 17, 2006) at 5, 27 -- 11/13/2006 14:37:12, 5, 27 -- Serialize complete at 11/13/2006 14:37:12 5, 27 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
I understand how and why to perform the diopter adjustment on the eyepieces of a light microscope but I must admit I don't get why this is called a "diopter adjustment". Can someone explain this terminology to me.
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Since the IC is an 8-connection IC, it is most likely not far from a "trivial" job to analyze it. First off, I think it is not a flip chip but rather a BGA. This is quite a different matter and makes analysis easier. If I am right, the die is on a lead frame and wire bonded to an internal substrate that is then solder bumped. Since the main board is expendable, then just cut out the bad IC (main PCB and all). Then heat to about 125F and cleave it off of the main PCB. I assume that the device is small in size. the other option is to grind off the main PCB. However, if it is assembled as I suspect, the circuit side of the IC is up, not down.
A quick x-ray of the device will show how it is constructed. A side x-ray will reveal the cross section detail of the die and package. From there, the plastic encapsulation can be removed with hot sulfuric or SF6 + O2 plasma. Alternatively, if the die is small, a focused ion beam can selectively expose the die. With an exposed die, reverse engineering can take place and then analyze areas for FA.
If the device is simply not connected well to the main PCB, then C-SAM analysis will show this.
If you are not into FA and reverse engineering, the tools to do this vastly exceed the value of knowing why the part failed, IMO. I suspect that if you return the module to the vendor, they will just say that it failed and offer a replacement module. If the failure happens frequently, I would still revert this issue to the module maker. In all likelihood, they designed the ASIC.
A close up pix of the device would be helpful.
gary g.
At 02:45 AM 11/13/2006, you wrote:
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==============================Original Headers============================== 16, 20 -- From gary-at-gaugler.com Mon Nov 13 15:28:28 2006 16, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 16, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kADLSSq7020755 16, 20 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 15:28:28 -0600 16, 20 -- Received: (qmail 28958 invoked from network); 13 Nov 2006 13:28:22 -0800 16, 20 -- Received: by simscan 1.1.0 ppid: 28935, pid: 28956, t: 0.3319s 16, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 16, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 16, 20 -- by qsmtp1 with SMTP; 13 Nov 2006 13:28:21 -0800 16, 20 -- Message-Id: {7.0.1.0.2.20061113131511.025c66f0-at-gaugler.com} 16, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 16, 20 -- Date: Mon, 13 Nov 2006 13:28:26 -0800 16, 20 -- To: richard.beanland-at-bookham.com 16, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 16, 20 -- Subject: Re: [Microscopy] RE: Failure analysis;flip-chip dies 16, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 16, 20 -- In-Reply-To: {200611131045.kADAj5Jb001196-at-ns.microscopy.com} 16, 20 -- References: {200611131045.kADAj5Jb001196-at-ns.microscopy.com} 16, 20 -- Mime-Version: 1.0 16, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
On Nov 13, 2006, at 12:58 PM, phillipst-at-missouri.edu wrote:
} I understand how and why to perform the diopter adjustment on the } eyepieces } of a light microscope but I must admit I don't get why this is called a } "diopter adjustment". Can someone explain this terminology to me. } Dear Thomas, A diopter is an inverse meter. (I think; the Handbook of Chemistry and Physics does not have the word in its glossary.) Lens power is measured in diopters, so the adjustment is just that power needed to have the eye pieces focus the image onto your retina. 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 Nov 13 16:03:39 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 kADM3d9i032118 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Nov 2006 16:03:39 -0600 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by water-ox-postvirus (Postfix) with ESMTP id 9D0202EEF1 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Nov 2006 14:03:38 -0800 (PST) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id 304D43548F 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 13 Nov 2006 14:03:32 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200611132058.kADKwdpk009817-at-ns.microscopy.com} 4, 22 -- References: {200611132058.kADKwdpk009817-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {8d1af803286af031668ed3568704015c-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] diopter adjustment 4, 22 -- Date: Mon, 13 Nov 2006 14:08:55 -0800 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==============================
I understand a diopter is the inverse of the focal length in meters (1/meter). I understand how a 2 diopter lens can combined with a 0.5 diopter lens to get a 2.5 diopter effect. but when you "focus" an eyepiece, aren't you simply raising or lowering the height of the ocular? I don't see how this is a diopter adjustment. Tom
At 04:04 PM 11/13/06, 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
I assume that you have checked, and tuned, the gun alignment with the fluorescent screen stub and made sure your flow rates are optimised (both of these are in the manual). Another possible one is if you are using a one sided milling stub and both your guns are angled to come from underneath, sounds silly but it can happen!
Matt
} Hi Everyone } } I have a problem with PIPS (Gatan) in our lab. The samples (including } metal and semiconductors) are not milled at all (like 10 hours at } voltage 4.5). We just cleaned the penning gauge and two guns. The vacuum } and current looks common. I wonder someone can give me suggestion what } is the problem. } } Thanks. } } Jiaming
-- Dr M.Weyland, Electron Microscopist
Monash Centre for Electron Microscopy Monash University Clayton Campus VIC 3800 Australia
==============================Original Headers============================== 7, 26 -- From mw275-at-cornell.edu Mon Nov 13 16:31:41 2006 7, 26 -- Received: from ALPHA1.ITS.MONASH.EDU.AU (alpha1.its.monash.edu.au [130.194.1.1]) 7, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADMVema021424 7, 26 -- for {Microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 16:31:41 -0600 7, 26 -- Received: from larry.its.monash.edu.au ([130.194.13.82]) 7, 26 -- by vaxc.its.monash.edu.au (PMDF V6.1 #31276) 7, 26 -- with ESMTP id {01M9JQ18FVWI91WNUK-at-vaxc.its.monash.edu.au} for 7, 26 -- Microscopy-at-microscopy.com; Tue, 14 Nov 2006 09:31:35 +1100 7, 26 -- Received: from larry.its.monash.edu.au (localhost.localdomain [127.0.0.1]) 7, 26 -- by localhost (Postfix) with ESMTP id 1D39480002 for 7, 26 -- {Microscopy-at-microscopy.com} ; Tue, 14 Nov 2006 09:31:35 +1100 (EST) 7, 26 -- Received: from [130.194.139.201] 7, 26 -- (dyn-130-194-139-201.eng.monash.edu.au [130.194.139.201]) 7, 26 -- (using TLSv1 with cipher DHE-RSA-AES256-SHA (256/256 bits)) 7, 26 -- (No client certificate requested) (Authenticated sender: mweyland) 7, 26 -- by larry.its.monash.edu.au (Postfix) with ESMTP id D884C3C006 for 7, 26 -- {Microscopy-at-microscopy.com} ; Tue, 14 Nov 2006 09:31:34 +1100 (EST) 7, 26 -- Date: Tue, 14 Nov 2006 09:31:40 +1100 7, 26 -- From: Matthew Weyland {mw275-at-cornell.edu} 7, 26 -- Subject: Re:[Microscopy] One problem with Ion milling 7, 26 -- To: Microscopy-at-microscopy.com 7, 26 -- Message-id: {4558F24C.6000605-at-cornell.edu} 7, 26 -- MIME-version: 1.0 7, 26 -- Content-type: text/plain; charset=ISO-8859-1; format=flowed 7, 26 -- Content-transfer-encoding: 7BIT 7, 26 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) ==============================End of - Headers==============================
We use DEVCON 5 minute epoxy to glue face-to-face, then use crazy bond (same as superglue, cyanoacrylate-base) for temporary fixture. Both gel-type or liquid type crazy bond worked fine for us. Acetone can be used to dissolve the bond. One word for DEVCON, it softens a little bit in Acetone, so don't put the ground piece too long time in acetone. M-bond will not be strong enough if it is not heated. Some others use UV-curing bonds as permanent gluing for some optical films and substrates. Good luck!
Young-Woon Kim Seoul National University School of Materials Science and Engineering Tel) +82-2-880-7977 Fax) +82-2-883-8197 e-mail) youngwk-at-snu.ac.kr
-----Original Message----- X-from: lkrupp-at-us.ibm.com [mailto:lkrupp-at-us.ibm.com] Sent: Tuesday, November 14, 2006 4:41 AM To: youngwk-at-snu.ac.kr
Hello-
I need to prepare a self supporting cross section of a multilayer film on a 1" silicon wafer. Problem is the material cannot be heated above room temperature. So as far as I know, I need to use something other than G-1 to prepare the sandwich and something other than crystalbond to do the polishing/dimpling. My first thought is M-bond 610 for the sandwich, I've heard this will cure overnight at RT? And superglue for the polishing, although this will require soaking off between switching sides. M-bond 610 is not soluble in acetone, right?
Any other ideas? Commercial responses are welcome.
The sample will be ion-milled at LN2 temperatures after the polishing/dimpling steps.
Thanks, Leslie _____________________ Leslie Krupp IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 5, 27 -- From lkrupp-at-us.ibm.com Mon Nov 13 13:37:14 2006 5, 27 -- Received: from e6.ny.us.ibm.com (e6.ny.us.ibm.com [32.97.182.146]) 5, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADJbEPS029873 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 13:37:14 -0600 5, 27 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 5, 27 -- by e6.ny.us.ibm.com (8.13.8/8.12.11) with ESMTP id kADJbXNw029806 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:33 -0500 5, 27 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) 5, 27 -- by d01relay02.pok.ibm.com (8.13.6/8.13.6/NCO v8.1.1) with ESMTP id kADJbDu5181762 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) 5, 27 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id kADJbD9N006725 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 5, 27 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id kADJbD4q006713 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- To: microscopy-at-microscopy.com 5, 27 -- MIME-Version: 1.0 5, 27 -- Subject: TEM sample prep: no heat adhesives required 5, 27 -- X-Mailer: Lotus Notes Release 7.0 HF277 June 21, 2006 5, 27 -- Message-ID: {OF98894D7C.38D025D7-ON85257225.006B12FC-88257225.006BC206-at-us.ibm.com} 5, 27 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 5, 27 -- Date: Mon, 13 Nov 2006 11:36:31 -0800 5, 27 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.2HF32 | October 17, 2006) at 5, 27 -- 11/13/2006 14:37:12, 5, 27 -- Serialize complete at 11/13/2006 14:37:12 5, 27 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
==============================Original Headers============================== 20, 28 -- From youngwk-at-snu.ac.kr Mon Nov 13 18:04:46 2006 20, 28 -- Received: from nabi1.snu.ac.kr (nabi1.snu.ac.kr [147.46.100.51]) 20, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAE04jeD001805 20, 28 -- for {Microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 18:04:45 -0600 20, 28 -- Received: from [147.46.100.31] ([147.46.100.31]) 20, 28 -- by nabi1.snu.ac.kr ([147.46.100.51]) 20, 28 -- with ESMTP id 2006111409:04:44:146893.21813.4849961 20, 28 -- for {Microscopy-at-microscopy.com} ; 20, 28 -- Tue, 14 Nov 2006 09:04:44 +0900 (KST) 20, 28 -- Received: from [147.46.233.36] ([147.46.233.36]) 20, 28 -- by auk1.snu.ac.kr ([147.46.100.31]) 20, 28 -- with ESMTP id 2006111409:04:37:580201.30788.353041328 20, 28 -- for {Microscopy-at-microscopy.com} ; 20, 28 -- Tue, 14 Nov 2006 09:04:37 +0900 (KST) 20, 28 -- From: "Young-Woon Kim" {youngwk-at-snu.ac.kr} 20, 28 -- To: {Microscopy-at-microscopy.com} 20, 28 -- Subject: RE: [Microscopy] TEM sample prep: no heat adhesives required 20, 28 -- Date: Tue, 14 Nov 2006 09:06:45 +0900 20, 28 -- Message-ID: {053501c70780$c58a1e00$24e92e93-at-young323} 20, 28 -- MIME-Version: 1.0 20, 28 -- Content-Type: text/plain; 20, 28 -- charset="us-ascii" 20, 28 -- Content-Transfer-Encoding: 7bit 20, 28 -- X-Mailer: Microsoft Office Outlook 11 20, 28 -- Thread-Index: AccHW5qRP/g/6POhSbiX7AjOtNlbuwAIf5JA 20, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 20, 28 -- X-TERRACE-SPAMMARK: NO (SR:22.71) 20, 28 -- (by Terrace) ==============================End of - Headers==============================
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Email: millerpod-at-hotmail.com Name: Andy Miller
Organization: Curtin Uni Western Australia
Title-Subject: [Filtered] phillips xl 30 quad current amplifier PCB
Question: Hi All
We have a problem with our Phillips XL 30.
The quad current amplifier PCB card that slots in the back was overheating and disrupts the scanning beam. We use a fan to cool it until we managed to get a replacement but that too had the same problem, which leads us to believe that it may not be a faulty card but a nearby component that is generating excess heat and in turn overheating the card.
Dose this sound feasible? Has anybody experienced this sort of problem? Could there be another reason aside from the remote chance that we got 2 dud cards?
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Email: t.keller-at-uni-jena.de Name: Thomas Keller
Organization: Institute of Materials Science / Friedrich-Schiller-University Jena
Title-Subject: [Filtered] Scheduling System
Question: Dear Kathy,
the Faces Scheduling System works fine:
http://faces.ccrc.uga.edu/
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perhaps just some clarification (arguments are not mine.....):
Definitions for:
DIOPTER: A measure of the optical power of a lens, equal to the reciprocal of its focal length in meters.
DIOPTER ADJUSTER: A mechanism in a light microscope eyepiece for adjusting the position of the eyelens to compensate for differences in dioptric power between the eyes of the observer(s)
DIOPTRIC: Regarding an optical system containing refractive elements
Reference: HEATH Julian P (Ed): Dictionary of Microscopy, (Micr.& Analysis) John WILEY & Sons LTD, ISBN-10:0-470-01199-8 (ISBN-13: 978-0-470-01199-7), 2005 pp 357
Assuming you are talking about adjustment of the ocular(s) according to a /your (refraction) myopy or hypermetropy:
I guess, you won't/don't adjust the } ocular's height { (by raising or lowering the "diopter adjustment ring") but instead "realiter" change the focal length of the ocular lens system to adapt the correct centering of } sample/vision rays { ON your retina instead of their otherwise crossing / centering before (myopy) or behind (metropy) the retinal cells. ==} "Adjust the diopter to suit the observer's eyesight. The method differs according to the eyepiece used. Unless the diopter is adjusted, parfocality will not be maintained when the objective is changed. (When using e.g. WHK 10x eyepieces, the focus is adjusted with the focusing knob[s] whil observing through the right eyepiece. The diopter adjustment ring on the left side is then adjusted for maximum image clarity for the left eye)......slightly different adjustment for "finder"-eyepieces (in photomicroscopy)......" Reference: How to improve Photography through the Microscope, Olympus Optical Co.Ltd. brochure (end of 1980ies; M132E-1191T)
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I understand a diopter is the inverse of the focal length in meters (1/meter). I understand how a 2 diopter lens can be combined with a 0.5 diopter lens to get a 2.5 diopter effect. but when you "focus" an eyepiece, aren't you simply raising or lowering the height of the ocular? I don't see how this is a diopter adjustment. Tom
At 04:04 PM 11/13/06, 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
I would like to use a thermoreversible gel (liquid at 4degree and under) to immobilise a multicellular aggregate to the membrane of a Quantomix capsule. This is then inserted in to a SEM.
Question, what is the internal temperature of a SEM?
Many thanks
==============================Original Headers============================== 6, 28 -- From Sam.Murray-at-port.ac.uk Tue Nov 14 08:20:51 2006 6, 28 -- Received: from betel.iso.port.ac.uk (betel.iso.port.ac.uk [148.197.254.2]) 6, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAEEKnfw031061 6, 28 -- for {Microscopy-at-Microscopy.Com} ; Tue, 14 Nov 2006 08:20:50 -0600 6, 28 -- Received: from ls04.nwservers.iso.port.ac.uk ([148.197.251.173]:14268) 6, 28 -- by betel.iso.port.ac.uk with smtp (Exim 4.60) 6, 28 -- (envelope-from {Sam.Murray-at-port.ac.uk} ) 6, 28 -- id 1Gjz9Y-0007Ki-IN 6, 28 -- for Microscopy-at-Microscopy.Com; Tue, 14 Nov 2006 14:20:44 +0000 6, 28 -- Received: from LS04/SpoolDir by ls04.nwservers.iso.port.ac.uk (Mercury 1.48); 6, 28 -- 14 Nov 06 14:20:44 +0000 6, 28 -- Received: from SpoolDir by LS04 (Mercury 1.48); 14 Nov 06 14:20:42 +0000 6, 28 -- From: "Samantha Murray" {Sam.Murray-at-port.ac.uk} 6, 28 -- Organization: University of Portsmouth 6, 28 -- To: Microscopy-at-Microscopy.Com 6, 28 -- Date: Tue, 14 Nov 2006 14:20:42 -0000 6, 28 -- MIME-Version: 1.0 6, 28 -- Subject: [Microscopy] Temperature inside SEM chamber? 6, 28 -- Message-ID: {4559D0BE.15056.1DBA01-at-localhost} 6, 28 -- X-Confirm-Reading-To: "Samantha Murray" {Sam.Murray-at-port.ac.uk} 6, 28 -- X-pmrqc: 1 6, 28 -- Return-receipt-to: "Samantha Murray" {Sam.Murray-at-port.ac.uk} 6, 28 -- Priority: normal 6, 28 -- X-mailer: Pegasus Mail for Windows (v4.12a) 6, 28 -- Content-type: text/plain; charset=US-ASCII 6, 28 -- Content-transfer-encoding: 7BIT 6, 28 -- Content-description: Mail message body 6, 28 -- X-Scan-Signature: 0fa7eb88a48416f360b5a84d0500e04d ==============================End of - Headers==============================
Either something is blocking the beams or, more likely, the beams are not aimed properly. Either problem is easy to diagnose as you can see the beams where they strike the sample if you darken the room. The sample holder should fluoresce by itself or paint a little aquadag on it. The beam aiming adjustments are discussed in the instruction book.
Ron Anderson
zhangj16-at-msu.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 Everyone } } I have a problem with PIPS (Gatan) in our lab. The samples (including } metal and semiconductors) are not milled at all (like 10 hours at } voltage 4.5). We just cleaned the penning gauge and two guns. The vacuum } and current looks common. I wonder someone can give me suggestion what } is the problem. } } Thanks. } } Jiaming } } } ==============================Original Headers============================== } 5, 16 -- From zhangj16-at-msu.edu Mon Nov 13 12:29:40 2006 } 5, 16 -- Received: from egr.msu.edu (jeeves.egr.msu.edu [35.9.37.127]) } 5, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADITeG3013018 } 5, 16 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 12:29:40 -0600 } 5, 16 -- Received: from [127.0.0.1] (muran.dhcp.egr.msu.edu [35.9.138.199]) } 5, 16 -- by egr.msu.edu (8.13.7/8.13.4) with ESMTP id kADITdGZ007932 } 5, 16 -- for {Microscopy-at-Microscopy.Com} ; Mon, 13 Nov 2006 13:29:41 -0500 (EST) } 5, 16 -- Message-ID: {4558B992.6030803-at-msu.edu} } 5, 16 -- Date: Mon, 13 Nov 2006 13:29:38 -0500 } 5, 16 -- From: Jiaming Zhang {zhangj16-at-msu.edu} } 5, 16 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) } 5, 16 -- MIME-Version: 1.0 } 5, 16 -- To: Microscopy-at-Microscopy.Com } 5, 16 -- Subject: One problem with Ion milling } 5, 16 -- Content-Type: text/plain; charset=GB2312 } 5, 16 -- Content-Transfer-Encoding: 7bit } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 5, 19 -- From randerson20-at-tampabay.rr.com Tue Nov 14 08:20:57 2006 5, 19 -- Received: from ms-smtp-02.tampabay.rr.com (ms-smtp-02.tampabay.rr.com [65.32.5.132]) 5, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAEEKucm031091 5, 19 -- for {Microscopy-at-Microscopy.Com} ; Tue, 14 Nov 2006 08:20:57 -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 (8.13.6/8.13.6) with ESMTP id kAEEKq3q029407; 5, 19 -- Tue, 14 Nov 2006 09:20:54 -0500 (EST) 5, 19 -- Message-ID: {4559D0C3.1010001-at-tampabay.rr.com} 5, 19 -- Date: Tue, 14 Nov 2006 09:20:51 -0500 5, 19 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 5, 19 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) 5, 19 -- MIME-Version: 1.0 5, 19 -- To: zhangj16-at-msu.edu, Listserver {Microscopy-at-Microscopy.Com} 5, 19 -- Subject: Re: [Microscopy] One problem with Ion milling 5, 19 -- References: {200611131829.kADITpAI013410-at-ns.microscopy.com} 5, 19 -- In-Reply-To: {200611131829.kADITpAI013410-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==============================
I'm just getting started with serious optical, SEM, and TEM work, but I _do_ have a bit of experience as an ion source designer (and sufferer), of nearly thirty years. I'm not familiar with the particular machine you are having trouble with, but it most likely uses a Kaufman low-energy-broad-beam ion source. I use them for other things, but the most frequent problem is that the two very delicate grids w/ all the tiny holes in them are shorted out by what is technically called 'crud' in the world of sourcery. There is a voltage potential between the two grids that extracts the ion plasma from within the source. It is easily diagnosed if you look at the current meter on the power supply, and see it is shorted out. The current draw will be essentially infinite, and the supply probably has an automatic cut-out to protect itself. If the grids are shorted, then that portion of the supply won't turn on. The remedy is dislodging the offending piece of 'crud' without just moving the problem to another spot. GENTLY try blowing a bit of nitrogen, (or any dry pure gas), over the face of the front grid into the source. If you are _extremely_ fortunate, it will dislodge the 'crud' and give you an open circuit between the two grids. If it doesn't work on the first try, try blowing through the grid from various angles and orientations. It is worth some patient effort, because taking the source apart is a delicate process, and gloves should _always_ be worn, to keep from contaminating the source with oil from fingerprints. There are probably some tiny, delicate, and very important parts involved, so this shouldn't be undertaken by anyone without considerable ion source experience, or you will probably cause even greater problems. On Tuesday, November 14, 2006, at 09:29AM, {randerson20-at-tampabay.rr.com} wrote: } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 2, 22 -- From ionsourcerer-at-mac.com Tue Nov 14 11:52:35 2006 2, 22 -- Received: from smtpout.mac.com (smtpout.mac.com [17.250.248.46]) 2, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAEHqWar025148 2, 22 -- for {Microscopy-at-Microscopy.Com} ; Tue, 14 Nov 2006 11:52:34 -0600 2, 22 -- Received: from mac.com (webmail040-S [10.13.128.40]) 2, 22 -- by smtpout.mac.com (Xserve/8.12.11/smtpout10/MantshX 4.0) with ESMTP id kAEHqP2G026524 2, 22 -- for {Microscopy-at-Microscopy.Com} ; Tue, 14 Nov 2006 09:52:25 -0800 (PST) 2, 22 -- Received: from webmail040 (localhost [127.0.0.1]) 2, 22 -- by mac.com (8.13.8/webmail040/MantshX 4.0) with ESMTP id kAEHqOn0024793 2, 22 -- for {Microscopy-at-Microscopy.Com} ; Tue, 14 Nov 2006 09:52:25 -0800 (PST) 2, 22 -- Date: Tue, 14 Nov 2006 09:52:24 -0800 2, 22 -- From: ionsourcerer {ionsourcerer-at-mac.com} 2, 22 -- To: Microscopy-at-Microscopy.Com 2, 22 -- Message-ID: {02BD3DE7-010E-1000-9762-5B3102984122-Webmail-10010-at-mac.com} 2, 22 -- Subject: Re: [Microscopy] Re: One problem with Ion milling 2, 22 -- MIME-Version: 1.0 2, 22 -- Content-Type: text/plain; charset=ISO-8859-1 2, 22 -- Content-Transfer-Encoding: 7bit 2, 22 -- X-Originating-IP: 72.93.106.133 2, 22 -- Received: from [72.93.106.133] from webmail.mac.com with HTTP; Tue, 14 Nov 2006 09:52:24 -0800 2, 22 -- X-Brightmail-Tracker: AAAAAA== 2, 22 -- X-Brightmail-scanned: yes ==============================End of - Headers==============================
Dear Leslie, When we were waiting for our M-610 to arrive, we used Devon 2-ton epoxy to make cross-section samples. It can be squeezed thin and is stronger than 5-minute epoxy, but takes longer to harden up than 5-minute. I think we left it overnight. Regards, Mary Mager Electron Microscopist Materials Eng. UBC #419 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA Phone: 604-822-5648 Fax: 604-822-3619 email: mager-at-interchange.ubc.ca
-----Original Message----- X-from: lkrupp-at-us.ibm.com [mailto:lkrupp-at-us.ibm.com] Sent: Monday, November 13, 2006 11:43 AM To: mager-at-interchange.ubc.ca
Hello-
I need to prepare a self supporting cross section of a multilayer film on a 1" silicon wafer. Problem is the material cannot be heated above room temperature. So as far as I know, I need to use something other than G-1 to prepare the sandwich and something other than crystalbond to do the polishing/dimpling. My first thought is M-bond 610 for the sandwich, I've heard this will cure overnight at RT? And superglue for the polishing, although this will require soaking off between switching sides. M-bond 610 is not soluble in acetone, right?
Any other ideas? Commercial responses are welcome.
The sample will be ion-milled at LN2 temperatures after the polishing/dimpling steps.
Thanks, Leslie _____________________ Leslie Krupp IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 5, 27 -- From lkrupp-at-us.ibm.com Mon Nov 13 13:37:14 2006 5, 27 -- Received: from e6.ny.us.ibm.com (e6.ny.us.ibm.com [32.97.182.146]) 5, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kADJbEPS029873 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 13:37:14 -0600 5, 27 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 5, 27 -- by e6.ny.us.ibm.com (8.13.8/8.12.11) with ESMTP id kADJbXNw029806 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:33 -0500 5, 27 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) 5, 27 -- by d01relay02.pok.ibm.com (8.13.6/8.13.6/NCO v8.1.1) with ESMTP id kADJbDu5181762 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) 5, 27 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id kADJbD9N006725 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 5, 27 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id kADJbD4q006713 5, 27 -- for {microscopy-at-microscopy.com} ; Mon, 13 Nov 2006 14:37:13 -0500 5, 27 -- To: microscopy-at-microscopy.com 5, 27 -- MIME-Version: 1.0 5, 27 -- Subject: TEM sample prep: no heat adhesives required 5, 27 -- X-Mailer: Lotus Notes Release 7.0 HF277 June 21, 2006 5, 27 -- Message-ID: {OF98894D7C.38D025D7-ON85257225.006B12FC-88257225.006BC206-at-us.ibm.com} 5, 27 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 5, 27 -- Date: Mon, 13 Nov 2006 11:36:31 -0800 5, 27 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.2HF32 | October 17, 2006) at 5, 27 -- 11/13/2006 14:37:12, 5, 27 -- Serialize complete at 11/13/2006 14:37:12 5, 27 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
==============================Original Headers============================== 12, 33 -- From mager-at-interchange.ubc.ca Tue Nov 14 13:57:28 2006 12, 33 -- Received: from mta3.mail-relay.ubc.ca (mta3.mail-relay.ubc.ca [137.82.45.6]) 12, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAEJvPti006035 12, 33 -- for {microscopy-at-microscopy.com} ; Tue, 14 Nov 2006 13:57:25 -0600 12, 33 -- Received: from mta1.interchange.ubc.ca (mta1.interchange.ubc.ca [142.103.145.69]) 12, 33 -- by mta3.mail-relay.ubc.ca (8.12.11.20060308/8.12.11) with ESMTP id kAEJvMkF004599 12, 33 -- for {microscopy-at-microscopy.com} ; Tue, 14 Nov 2006 11:57:22 -0800 (PST) 12, 33 -- (envelope-from mager-at-interchange.ubc.ca) 12, 33 -- Received: from Mary (desk.staff.mmat.ubc.ca [137.82.16.178]) 12, 33 -- by smtp.interchange.ubc.ca 12, 33 -- (iPlanet Messaging Server 5.2 HotFix 1.21 (built Sep 8 2003)) 12, 33 -- with ESMTPA id {0J8Q00JD0KRLBG-at-smtp.interchange.ubc.ca} for 12, 33 -- microscopy-at-microscopy.com; Tue, 14 Nov 2006 11:57:22 -0800 (PST) 12, 33 -- Date: Tue, 14 Nov 2006 11:57:51 -0800 12, 33 -- From: Mary Mager {mager-at-interchange.ubc.ca} 12, 33 -- Subject: RE: [Microscopy] TEM sample prep: no heat adhesives required 12, 33 -- In-reply-to: {200611131942.kADJgoK9006665-at-ns.microscopy.com} 12, 33 -- To: lkrupp-at-us.ibm.com 12, 33 -- Cc: Microscopy {microscopy-at-microscopy.com} 12, 33 -- Reply-to: mager-at-interchange.ubc.ca 12, 33 -- Message-id: {0J8Q00JD1KRMBG-at-smtp.interchange.ubc.ca} 12, 33 -- Organization: Materials Eng. UBC 12, 33 -- MIME-version: 1.0 12, 33 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2800.1478 12, 33 -- X-Mailer: Microsoft Office Outlook, Build 11.0.6353 12, 33 -- Content-type: text/plain; charset=us-ascii 12, 33 -- Content-transfer-encoding: 7bit 12, 33 -- Thread-index: AccHW+imBfW78FlLRsWHk+se/AhqigAytzDA 12, 33 -- X-UBC-Scanned: Sophos PureMessage 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.11.14.105432 12, 33 -- X-UBC-Relayed: Relayed through mail-relay.ubc.ca 12, 33 -- X-PerlMx-Spam: Probability=37%, Report=OBFU_CLASS_OFFENSIVE_MED 4, __C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_PHRASE_24 0, __USER_AGENT_MS_GENERIC 0 12, 33 -- X-Spam-Level: XXX 12, 33 -- X-Spam-Flag: No ==============================End of - Headers==============================
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America I guess you actually mean the temperature under the beam ..... The temperature in the specimen chamber generally is going to be close to ambient (except for an ESEM, were the sample will be ~5 deg C)
Key things to consider are probe current, scan speed, magnification and thermal conduction path away from the imaged area. In the case of Quantomix capsules, with a FEG-SEM it is relatively easy to punch holes in the membrane with the beam, rather less so with W SEMs.
It is also relatively easy in most SEMs to find conditions which will melt lower temperature thermosetting polymers, so local sample temperatures in the 50 to 100 deg C aren't too difficult to generate, with the right sample.
Basically, I think there are so many variables involved, you're just going to have to try it.
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============================== 6, 16 -- From larry-at-celtic.freewire.co.uk Tue Nov 14 15:04:26 2006 6, 16 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 6, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAEL4Nue017767 6, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 14 Nov 2006 15:04:26 -0600 6, 16 -- Received: from [194.164.78.253] (modem-rack4-modem253.netkonect.net [194.164.78.253]) 6, 16 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id kAEL2b3B004597; 6, 16 -- Tue, 14 Nov 2006 21:03:31 GMT 6, 16 -- Mime-Version: 1.0 6, 16 -- Message-Id: {p06210202c17fd7b6ceb4-at-[194.164.78.26]} 6, 16 -- In-Reply-To: {200611141429.kAEETu5l019725-at-ns.microscopy.com} 6, 16 -- References: {200611141429.kAEETu5l019725-at-ns.microscopy.com} 6, 16 -- Date: Tue, 14 Nov 2006 20:38:18 +0000 6, 16 -- To: Sam.Murray-at-port.ac.uk, Microscopy-at-MSA.Microscopy.Com 6, 16 -- From: Larry Stoter {larry-at-celtic.freewire.co.uk} 6, 16 -- Subject: Re: [Microscopy] Temperature inside SEM chamber? 6, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
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Email: pbgrover-at-yahoo.com Name: Paul Grover
Organization: Purdue University
Title-Subject: [Filtered] integrity in scientific research
Question: Dear Microscopists,
I find it difficult to understand, based on the responses, both on and = offlist, to the thread on research ethics back in April, that no one has = yet posted to the forum I recently set up (Yahoo group 'honestscience'). = Could this be because of concerns over anonymity? If so, please = contact me or someone else to find out how to post using an alias. = Sorry to waste more listserver space, but since quite a few people said = they would welcome such a forum, I just don't understand why there = hasn't been any activity. Best regards to you all.
Paul Grover, Ph.D. Owner, Grover Roofing and Remodeling and Chief Microscopist and Bottle Washer, Microvista Laboratory
After a computer crash and a restart of the system, I found that the beam alignment was much worse than before. I tried reloading previous alignment settings, but with no effect. The main problem is that when I change the magnification, the beam shifts, especially between the mag 44kx and 55kx. I did once the full alignment procedure, and I don't want to repeat it again (and then it is not necessary). I just need to know how to correct this specific problem, either using the direct alignment or within the alignment section. I tried centering the beam with the multifunction knobs using direct alignment/beam shift, saving the alignment for each magnification but it does not fix the problem. Does anyone have any idea?
Stephane
==============================Original Headers============================== 6, 19 -- From nizets2-at-yahoo.com Wed Nov 15 08:53:48 2006 6, 19 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.91.145]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAFErmx4030210 6, 19 -- for {microscopy-at-microscopy.com} ; Wed, 15 Nov 2006 08:53:48 -0600 6, 19 -- Received: (qmail 98133 invoked by uid 60001); 15 Nov 2006 14:53:48 -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:MIME-Version:Content-Type:Content-Transfer-Encoding; 6, 19 -- b=xnjd5drUu6sCGIGWFylc1gLavaxo/Ol50NhMMvmfvACvFxMpxxQVlDWA7ME905Kd2jDnm68OuWCo2xtZQFDDBr13561Rd/6NU9ySYJtuK3OXgwtLSa77s5S4/TedzoeSxVchaXkD9Vd+Ps/Sv4dcLhVjqrCraxEFcoJQTJiOvCQ= ; 6, 19 -- Message-ID: {20061115145348.98131.qmail-at-web37413.mail.mud.yahoo.com} 6, 19 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Wed, 15 Nov 2006 06:53:48 PST 6, 19 -- Date: Wed, 15 Nov 2006 06:53:48 -0800 (PST) 6, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 19 -- Subject: alignment with tecnai twin G20 6, 19 -- To: microscopy-at-microscopy.com 6, 19 -- MIME-Version: 1.0 6, 19 -- Content-Type: text/plain; charset=ascii 6, 19 -- Content-Transfer-Encoding: 8bit 6, 19 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAFErmx4030210 ==============================End of - Headers==============================
I do not know how FEI tell you to align the lenses but set out below are generic lens alignment procedures - after all FEI produce a TEM so TEM procedures should work!
1. Switch between the two magnifications that are proving to be a problem. 2. Check the lens currents to find which lenses are activated during this change 3 If an intermediate lens look at alignment A if a projector lens look at alignment B.
A
A1 When switching the diffraction spot will flash on the screen. A2 Adjust the intermediate lens such that when switching in either direction the diffraction spot appears in exactly the same area, this may not be on the centre of the screen. A3 Adjust the lens above the problem lens to centre this diffraction spot on the screen. A4 Re check from A2.
B
B1 At the lower magnification align the first projector lens image centre on the centre of the screen. B2 Switch to the higher magnification and centre the second projector image centre on the centre of the screen. B3 Repeat until centres are exactly the same.
Hope this helps?
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: {nizets2-at-yahoo.com} To: {protrain-at-emcourses.com} Sent: Wednesday, November 15, 2006 2:54 PM
Hi,
If you mean "mechanical" adjustments of the lenses: you can't do this on a Tecnai.
Are you sure this shift wasn't always happening?
Regards, Sander
-----Original Message----- X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com] Sent: Wednesday, November 15, 2006 7:27 PM To: Stoks, Sander
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Email: j.janssen-at-nki.nl Name: Hans Janssen
Organization: Netherlands Cancer Institute
Title-Subject: [Filtered] low contrast on uranyl staining of tokoyasu sections
Question: For some time now we have problems with low contrast on our tokoyasu sections for immunoEM. After immune incubation we stain for 5 minutes with uranyloxalate pH 7, wash in H2O, followed by 0.6% uranylacetate in methylcellulose. This used to give a great contrast, but now membranes are hardly visible anymore. We tried solving this by using different brands and batches and higher percentages, but so far it did not get better. Maybe I open up an old discussion, but I am new to the listserver, so sorry in that case. Hope somebody can give some new insights.
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Email: pocohe-at-yahoo.com Name: Yong He
Organization:
Title-Subject: [Filtered] instruction for off-axis electron holography
Question: Dear lister, I am trying to research some magnetic materials by off-axis electron holography, but I am a new hand for using electron holography and still want to try it step by step if I can get one manual for electron holography. I will appreciate you if you can send me one copy by email.
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Email: wa5ekh-at-juno.com Name: charles Jeffrey Day
Question: The company I work for in N. Texas has a surplus E3 Electroscan ESEM, 19= 89. When they have decided to replace it with a new instrument, they qui= t repairing this instrument. The ESEM probably has a vacuum guage board = that needs repair and the EDS(Tracor Noran Voyager,Be Window, 1989) also= has a board issue. =
ALSO...I'll tell you working with water(vapor) detection at 1-20 torr ha= s been a complete surprise, or revelation!, to an older EM operator (mys= elf), who was for years been indoctrinated in "High Vac/High Voltage/LOW= Water Vapor,Low Hydrocarbon" EM technology. There is a great deal more = to this "Environmental EM" than I believe we realize. Not just the abili= ty to image uncoated samples! For instance "STEEM" (my favorite personal= term for Transmission ESEM) techniques for Ultrathin Cross Section imag= ing at 20 KV is very interesting example! Have you seen?? ....images fro= m HRETEM? I'd like to hear more from this community. You can contact me at 817-792-1434 (direct or voice mail). Jeff Day wa5ekh-at-juno.com (Charles Jeffrey Day) (note: I receive no personal profit from this.)
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Title-Subject: [Filtered] Entry to Mid Level Dual Beam Operator
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Email: smythen-at-musc.edu Name: Nancy Smythe
Organization: Medical University of South Carolina
Title-Subject: [Filtered] Uranyl Acetate staining
Question: We also had the problem several months ago and not just in my lab but the core facility at the university alse. We found, after much frustration, that the older the UA the better it works. Must be something in the processing of the newer batches that is causing this. Lucky for me I found some in the back of the cabinet that will keep me going for a long time.
We've got 5g of UA here, NOS unopened and dating to 1995, if I am reading the faded ink note correctly. It's free to anyone who can arrange for and pay for shipping.
Ron L
-----Original Message----- X-from: smythen-at-musc.edu [mailto:smythen-at-musc.edu] Sent: Thursday, November 16, 2006 9:29 AM To: lherault-at-bu.edu
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Email: smythen-at-musc.edu Name: Nancy Smythe
Organization: Medical University of South Carolina
Title-Subject: [Filtered] Uranyl Acetate staining
Question: We also had the problem several months ago and not just in my lab but the core facility at the university alse. We found, after much frustration, that the older the UA the better it works. Must be something in the processing of the newer batches that is causing this. Lucky for me I found some in the back of the cabinet that will keep me going for a long time.
I found the same product as a zinc salt in Sigma (it is written "tetraphenylprophinE) I don't think silver is more toxic than zinc. Actually the product does not seem to be very dangerous. I don't know why the MSDS by Sigma is in french, but I can translate it for you since french is my mother language. I know that this is no sulfanate, but without CAS number, catalog number or molecular weight this is all I could find.
Disposal of the substance Dissolve or mix the product with a solvant and burn in an appropriate incinerator (appropriate for burning chimicals). Please follow the local rules (this is the sadly usual and helpless answer you ever get from Sigma when you ask how to dispose of a product. They sell them, and don't care what they become afterwards).
Hope this helped.
regards,
Stephane
----- Original Message ---- X-from: "DES-at-CDC.GOV" {DES-at-CDC.GOV} To: nizets2-at-yahoo.com Sent: Thursday, November 16, 2006 3:30:43 PM
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Email: DES-at-CDC.GOV Name: Diane Schwegler-Berry
Organization: NIOSH/CDC
Title-Subject: [Filtered] MSDS
Question: I have an old container of Silver tetraphenylporphin sulfanate and wish to dispose of it. I am looking for a MSDS for this chemical. Does anyone have a copy? Thank you.
unfortunately I don't have an MSDS in my relatively huge collection of....and, as you have perhaps also found, there are only a few article links for the stuff in the web online available, no hint for an MSDS....anyway...
Two suggestions (not based on a real "chemical knowledge" of the chemical, but comparing with substances like silver proteinate....SPI product info sheet states "not hazardous in terms of transportation"):
Perhaps either there would be a possibility of recycling the silver (cf. websource } Tape Data Recovery at Ilford { at http://www.data-recovery-london.com/Tape-Data-Recovery-at-Ilford.html , quoting also Silver-tetraphenylporphinsulfonate....but unfortunately there is no free access to the underlying data collection), or, to talk to your safety officer(s) concerning safe disposal (if you do have "analog"/"ancillary" photographic waste solutions perhaps you would like to add the substance to the hydrous waste solution of fixator ....?....but I not quite sure about it....)....
Hoping there is better help out there.... Good luck and best wishes
Wolfgang MUSS Salzburg-Austria
PS: perhaps you can use information by WIKIPEDIA, to evaluate a possible "toxicity" of the substance, searching e.g. for "porphine" and/or "pyrrole" (in the German Wikipedia I found (translated): Porphine, heterocyclic (contains 4 N-atoms), aromatic chemical substance, consisting of 4 Pyrrol-rings (==} Tetrapyrrol)...the Tetrapyrrol-ring forms the basic skeleton of the Haem, of all porphyrins and of chlorophylls....(so perhaps only "phenyl" is left to be examined for a toxicity.....) Sulfonic acids are strong acids, acids and sulfonates mostly are watersoluble and relativley well miscible with water).....
hope that adds some info valuable for getting rid of your problem.....
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Question: I have an old container of Silver tetraphenylporphin sulfanate and wish to dispose of it. I am looking for a MSDS for this chemical. Does anyone have a copy? Thank you.
MSDSonline - UCSF MSDS Management System http://www.msdsonline.com/accessaccountsearch/default.aspx?id=1000002
Iowa State University - Database of Material Safety Data Sheets http://www.ehs.iastate.edu/isumsds/ehsmsds.asp
Cornell University - Database of Material Safety Data Sheets http://msds.ehs.cornell.edu/msdssrch.asp
Arkansas State University - Database of Material Safety Data Sheets http://ehc.astate.edu/
Howard Hughes Medical Institute - Laboratory Chemical Safety Summaries http://www.hhmi.org/science/labsafe/lcss/index.html
US Environmental Protection Agency - Envirofacts Warehouse Chemical References Index http://www.epa.gov/enviro/html/emci/chemref/index.html
Micromedex Health Series - Summaries & detailed monographs for drugs, toxicological managements, & more.... http://www.thomsonhc.com/hcs/librarian/PFPUI/MY4CzEP1mg4dfj
Macquarie University - Material Safety Data Sheet Information http://www.chem.mq.edu.au/links.html#safety
University of Akron - Hazardous Chemical Database http://ull.chemistry.uakron.edu/erd/
The Vermont SIRI - Collection of Material Safety Data Sheets http://siri.org/msds/
Oxford University - Database of Material Safety Data Sheets http://physchem.ox.ac.uk/MSDS/
Sigma-Aldrich - Database of Material Safety Data Sheets http://www.sigmaaldrich.com/Area_of_Interest/The_Americas/United_States.html
-- 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============================== 19, 30 -- From Larry.Ackerman-at-ucsf.edu Thu Nov 16 11:51:28 2006 19, 30 -- Received: from emfmcb02.ucsfmedicalcenter.org (EMFMCB02.ucsfmedicalcenter.org [64.54.46.98]) 19, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAGHpRtF032384 19, 30 -- for {Microscopy-at-microscopy.com} ; Thu, 16 Nov 2006 11:51:28 -0600 19, 30 -- Received: from 64.54.128.153 by emfmcb02.ucsfmedicalcenter.org with 19, 30 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 19, 30 -- Thu, 16 Nov 2006 10:03:40 -0800 19, 30 -- X-Server-Uuid: 44B63953-633F-4500-B2DA-A3E478718104 19, 30 -- Received: from [128.218.123.88] ([128.218.123.88]) by 19, 30 -- exvs06.net.ucsf.edu with Microsoft SMTPSVC(6.0.3790.1830); Thu, 16 Nov 19, 30 -- 2006 09:51:22 -0800 19, 30 -- Message-ID: {455CA513.4020805-at-ucsf.edu} 19, 30 -- Date: Thu, 16 Nov 2006 09:51:15 -0800 19, 30 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 19, 30 -- Reply-to: larry.ackerman-at-ucsf.edu 19, 30 -- Organization: UCSF, NeuroAnatomy 19, 30 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 19, 30 -- X-Accept-Language: en-us, en 19, 30 -- MIME-Version: 1.0 19, 30 -- To: DES-at-CDC.GOV, Microscopy-at-microscopy.com 19, 30 -- Subject: Re: [Microscopy] MSDS 19, 30 -- References: {200611161430.kAGEUH1q010255-at-ns.microscopy.com} 19, 30 -- In-Reply-To: {200611161430.kAGEUH1q010255-at-ns.microscopy.com} 19, 30 -- X-OriginalArrivalTime: 16 Nov 2006 17:51:22.0482 (UTC) 19, 30 -- FILETIME=[D407D520:01C709A7] 19, 30 -- X-WSS-ID: 694278761IO4718182-01-01 19, 30 -- Content-Type: text/plain; 19, 30 -- charset=iso-8859-1; 19, 30 -- format=flowed 19, 30 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
Greetings all, Does anyone happen to know what the numerical aperture of the c-mount standard for light microscopy is? Thanks in advance. Best Regards, Karl
-- Karl Garsha Head Applications Scientist Roper Bioscience 3440 E. Brittania Drive Tucson, AZ 85706 Office: 520-547-2704
==============================Original Headers============================== 3, 27 -- From garsha-at-itg.uiuc.edu Thu Nov 16 16:39:36 2006 3, 27 -- Received: from outmail129167.authsmtp.com (outmail129167.authsmtp.com [62.13.129.167]) 3, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAGMdY5m016846 3, 27 -- for {microscopy-at-microscopy.com} ; Thu, 16 Nov 2006 16:39:36 -0600 3, 27 -- Received: from outmail128183.authsmtp.com (outmail128183.authsmtp.com [62.13.128.183]) 3, 27 -- by punt3.authsmtp.com (8.13.8/8.13.8/Kp) with ESMTP id kAGMdXq2029374 3, 27 -- for {microscopy-at-microscopy.com} ; Thu, 16 Nov 2006 22:39:33 GMT 3, 27 -- Received: from [144.22.2.171] (66-162-43-45.static.twtelecom.net [66.162.43.45]) 3, 27 -- (authenticated bits=0) 3, 27 -- by mail.authsmtp.com (8.13.6/8.13.6/Kp) with ESMTP id kAGMdRXZ088059 3, 27 -- for {microscopy-at-microscopy.com} ; Thu, 16 Nov 2006 22:39:28 GMT 3, 27 -- Message-ID: {455CE89B.6000601-at-itg.uiuc.edu} 3, 27 -- Date: Thu, 16 Nov 2006 15:39:23 -0700 3, 27 -- From: Karl Garsha {garsha-at-itg.uiuc.edu} 3, 27 -- Reply-To: kgarsha-at-roperscientific.com 3, 27 -- Organization: Roper Scientific 3, 27 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) 3, 27 -- MIME-Version: 1.0 3, 27 -- To: microscopy-at-microscopy.com 3, 27 -- Subject: NA or F# of C-Mount Standard 3, 27 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 3, 27 -- Content-Transfer-Encoding: 7bit 3, 27 -- X-Server-Quench: 51cc26a1-75c3-11db-b4dc-001185d377ca 3, 27 -- X-AuthRoute: OCdyaAgTClZaRR4B CiosDTNPDxgkOxYK DBMeOw5bK0AOTg9W KldyK1tYKloHTlZB SnhYBAkaUVtuJzwz dAhRbQNNYEhEXAxg UkAHRFRMFQNqHxgC GBobTRt3dQBZeDAr FDEfChIzNkRyfUZ4 SwBTFWpIYGYzO2IZ AUFFcAYHcx5Lf0sX dwJ7ACAQYWUGZ3Jl E1RsYDs4KxN2YCNP Ci9GZVUcCW1OOzkh QQxKJikmG0EMXSlb 3, 27 -- X-Authentic-SMTP: 61633138303639.squirrel.dmpriest.net.uk:315/Kp 3, 27 -- X-Report-SPAM: If SPAM / abuse - report it at: http://www.authsmtp.com/abuse 3, 27 -- X-Virus-Status: No virus detected - but ensure you scan with your own anti-virus system! ==============================End of - Headers==============================
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Title-Subject: [Filtered] particle tracking and stage movement
Question: I am trying to find out how to track particles under a microscope, and use the particle co-ordinates to reset the stage position (so that I can follow it over long periods of time). We use metamorph to run our microscope and stage, and ideally, I am looking for a solution using this software. Thanks!
Steven Lee Chief Technologist Electron Microscopy Laboratory Baylor University Medical Center 3500 Gaston Ave Dallas, TX 75246 Ph: 214.820.3302 Fx: 214.820.4110 Em: stevenle-at-baylorhealth.edu
-----Original Message----- X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at] Sent: Thursday, November 16, 2006 10:11 AM To: Lee, Steven
Dear Diane,
unfortunately I don't have an MSDS in my relatively huge collection of....and, as you have perhaps also found, there are only a few article links for the stuff in the web online available, no hint for an MSDS....anyway...
Two suggestions (not based on a real "chemical knowledge" of the chemical, but comparing with substances like silver proteinate....SPI product info
sheet states "not hazardous in terms of transportation"):
Perhaps either there would be a possibility of recycling the silver (cf. websource } Tape Data Recovery at Ilford { at http://www.data-recovery-london.com/Tape-Data-Recovery-at-Ilford.html , quoting also Silver-tetraphenylporphinsulfonate....but unfortunately there is no free access to the underlying data collection), or, to talk to your safety officer(s) concerning safe disposal (if you do have "analog"/"ancillary" photographic waste solutions perhaps you would
like to add the substance to the hydrous waste solution of fixator ....?....but I not quite sure about it....)....
Hoping there is better help out there.... Good luck and best wishes
Wolfgang MUSS Salzburg-Austria
PS: perhaps you can use information by WIKIPEDIA, to evaluate a possible
"toxicity" of the substance, searching e.g. for "porphine" and/or "pyrrole" (in the German Wikipedia I found (translated): Porphine, heterocyclic (contains 4 N-atoms), aromatic chemical substance, consisting of 4 Pyrrol-rings (==} Tetrapyrrol)...the Tetrapyrrol-ring forms the basic skeleton of the Haem, of all porphyrins and of chlorophylls....(so perhaps only "phenyl" is left to be examined for a toxicity.....) Sulfonic acids are strong acids, acids and sulfonates mostly are watersoluble and relativley well miscible with water).....
hope that adds some info valuable for getting rid of your problem.....
---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
---- This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html ------------------------------------------------------------------------ --- Remember this posting is most likely not from a Subscriber, so when replying please copy both DES-at-CDC.GOV as well as the MIcroscopy Listserver ------------------------------------------------------------------------ --- Email: DES-at-CDC.GOV Name: Diane Schwegler-Berry Organization: NIOSH/CDC Title-Subject: [Filtered] MSDS
Question: I have an old container of Silver tetraphenylporphin sulfanate
and wish to dispose of it. I am looking for a MSDS for this chemical. Does anyone have a copy? Thank you.
==============================Original Headers============================== 6, 12 -- From zaluzec-at-microscopy.com Thu Nov 16 08:25:16 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 kAGEPFBO027426 6, 12 -- for {microscopy-at-microscopy.com} ; Thu, 16 Nov 2006 08:25:16 -0600 6, 12 -- Mime-Version: 1.0 6, 12 -- X-Sender: (Unverified) 6, 12 -- Message-Id: {p0611040bc182253462f9-at-[206.69.208.22]} 6, 12 -- Date: Thu, 16 Nov 2006 08:25:14 -0600 6, 12 -- To: microscopy-at-microscopy.com 6, 12 -- From: DES-at-CDC.GOV (by way of MicroscopyListserver) 6, 12 -- Subject: viaWWW: MSDS Silver tetraphenylporphin 6, 12 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
==============================Original Headers============================== 24, 39 -- From W.Muss-at-salk.at Thu Nov 16 10:03:20 2006 24, 39 -- Received: from hermes.salk.at (hermes.lks.at [193.170.167.9]) 24, 39 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAGG3JAp017738 24, 39 -- for {microscopy-at-microscopy.com} ; Thu, 16 Nov 2006 10:03:19 -0600 24, 39 -- Received: from localhost (localhost [127.0.0.1]) 24, 39 -- by hermes.salk.at (Postfix) with ESMTP id 8F7E7C387E; 24, 39 -- Thu, 16 Nov 2006 17:03:16 +0100 (CET) 24, 39 -- X-Virii-Scanned: Kaspersky Antivirus at salk.at 24, 39 -- Received: from hermes.salk.at ([127.0.0.1]) 24, 39 -- by localhost (n1ex218.lks.local [127.0.0.1]) (amavisd-new, port 10024) 24, 39 -- with ESMTP id oaQ9yAAWv++s; Thu, 16 Nov 2006 17:03:16 +0100 (CET) 24, 39 -- Received: from hermes.lks.at (hermes.salk.at [192.168.13.210]) 24, 39 -- by hermes.salk.at (Postfix) with ESMTP id 1878CC387A; 24, 39 -- Thu, 16 Nov 2006 17:03:16 +0100 (CET) 24, 39 -- Received: from localhost (localhost [127.0.0.1]) 24, 39 -- by hermes.lks.at (Postfix) with ESMTP id ED1985A9048; 24, 39 -- Thu, 16 Nov 2006 17:03:15 +0100 (CET) 24, 39 -- Received: from hermes.lks.at ([127.0.0.1]) 24, 39 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 24, 39 -- with ESMTP id 92792-06; Thu, 16 Nov 2006 17:03:15 +0100 (CET) 24, 39 -- Received: from c1pa003 (unknown [192.168.42.3]) 24, 39 -- by hermes.lks.at (Postfix) with SMTP id 4BD1D5A9044; 24, 39 -- Thu, 16 Nov 2006 17:03:15 +0100 (CET) 24, 39 -- Received: by localhost with Microsoft MAPI; Thu, 16 Nov 2006 17:03:11 +0100 24, 39 -- Message-ID: {01C709A1.18A1F3C0.W.Muss-at-salk.at} 24, 39 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 24, 39 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 24, 39 -- To: "'DES-at-CDC.GOV'" {DES-at-CDC.GOV} 24, 39 -- Cc: "'microscopy-at-microscopy.com'" {microscopy-at-microscopy.com} 24, 39 -- Subject: [Microscopy] Re: MSDS Silver tetraphenylporphinsulfonate (S-TPPS) 24, 39 -- Date: Thu, 16 Nov 2006 17:03:09 +0100 24, 39 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 24, 39 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 24, 39 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 24, 39 -- MIME-Version: 1.0 24, 39 -- Content-Type: text/plain; charset="us-ascii" 24, 39 -- Content-Transfer-Encoding: 7bit 24, 39 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at 24, 39 -- X-Scanned-By: SALK-Content-Filter ==============================End of - Headers==============================
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==============================Original Headers============================== 41, 25 -- From StevenLe-at-BaylorHealth.edu Fri Nov 17 06:24:11 2006 41, 25 -- Received: from BHDAEXIMS01.bhcs.pvt (mailhost1.baylorhealth.edu [65.248.93.160]) 41, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAHCOAsc022852 41, 25 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Nov 2006 06:24:11 -0600 41, 25 -- Received: from BHDAEXFE01.bhcs.pvt ([10.5.3.72]) by BHDAEXIMS01.bhcs.pvt with InterScan Messaging Security Suite; Fri, 17 Nov 2006 06:24:10 -0600 41, 25 -- Received: from BHDAEXCH11.bhcs.pvt ([10.5.3.67]) by BHDAEXFE01.bhcs.pvt with Microsoft SMTPSVC(6.0.3790.211); 41, 25 -- Fri, 17 Nov 2006 06:24:11 -0600 41, 25 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 41, 25 -- Content-class: urn:content-classes:message 41, 25 -- MIME-Version: 1.0 41, 25 -- Content-Type: text/plain; 41, 25 -- charset="US-ASCII" 41, 25 -- Subject: RE: [Microscopy] Re: MSDS Silver tetraphenylporphinsulfonate (S-TPPS) 41, 25 -- Date: Fri, 17 Nov 2006 06:24:09 -0600 41, 25 -- Message-ID: {74A8B7A904152141BD5AA2761F5D223401FF9D78-at-BHDAEXCH11.bhcs.pvt} 41, 25 -- In-Reply-To: {200611161611.kAGGB54L027323-at-ns.microscopy.com} 41, 25 -- X-MS-Has-Attach: 41, 25 -- X-MS-TNEF-Correlator: 41, 25 -- Thread-Topic: [Microscopy] Re: MSDS Silver tetraphenylporphinsulfonate (S-TPPS) 41, 25 -- Thread-Index: AccJmeULrNy1zzVGRb+UE6aI/f8hagAqSk3g 41, 25 -- From: "Lee, Steven" {StevenLe-at-BaylorHealth.edu} 41, 25 -- To: {Microscopy-at-microscopy.com} 41, 25 -- X-OriginalArrivalTime: 17 Nov 2006 12:24:11.0967 (UTC) FILETIME=[49BEB4F0:01C70A43] 41, 25 -- Content-Transfer-Encoding: 8bit 41, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAHCOAsc022852 ==============================End of - Headers==============================
------------------------------------------------------------------------ ------------------------------------------------------------------------ ------------------------------------------------------------------------ ---------- The Italian Society for Pure and Applied Biophysics (SIBPA http://sibpa.itc.it/) in collaboration with: Istituto Veneto di Scienze Lettere ed Arti (IVSLA)
proudly announces the
XI SCHOOL OF PURE AND APPLIED BIOPHYSICS Advanced Optical Microscopy Methods in Biophysics
Venice, January 29 to February 2 - 2007, Palazzo Franchetti, the prestigious premises of the Istituto di Scienze Lettere e Arti (IVSLA) (http://www.ivsla.it).
Scientific Coordinators
* Giovanni Giacometti (IVSLA and University of Padova) * Giovanni Felice Azzone (IVSLA and University of Padova) * Alberto Diaspro (Dept. of Physics, University of Genova) * Cesare Usai (Institue of Biophysics, CNR)
Director of the School
* Giorgio M. Giacometti (IVSLA and University of Padova) E-mail: gcometti(at)bio.unipd.it
School overview and program Biophysics is a molecular science rapidly moving to the nanoscale. It seeks to explain biological function in terms of the molecular structures and properties of specific molecules. The size of these molecules varies dramatically, from small fatty acids and sugars (~1 nm = 10-9 m), to macromolecules like proteins (5-10 nm), starches (bigger than 1000 nm), and the enormously elongated DNA molecules. Much effort in biophysics is directed to determining the structure of specific biological molecules and of the larger structures into which they assemble. Some of this effort involves inventing new methods and building new instruments for monitoring these structures, and many of the exciting new developments in optical microscopy, in terms of imaging and manipulation, spectroscopy and visualization, are part of this effort.
The School is co-organized by Alberto Diaspro (Department of Physics, University of Genoa) and Cesare Usai (Institute of Biophysics, National Research Council). Topics of the School will be on: CARS (Coherent Anti Raman Scattering), TIRF (Total Internal Reflection Fluorescence), SHG-THG (Second-Third Harmonic Generation), Correlative microscopy, Multiphoton microscopy, Confocal Microscopy, 3D microscopy, Lifetime Imaging Microscopy, FRET (Forster resonance Enrgy Transfer), FRAP (Fluorescence Recovery After Photobleaching), SIngle molecule imaging, Tissue imaging, Cell imaging, Inverse problems in Optical Microscopy (Computational methods in image recovery), 7D Microscopy (adding new dimensions to x-y-z-t), Nanoscopy, Optical Tweezers, Complementary methods, Fluorescence Spectroscopy.
Provisional list of Lecturers includes:
* Wolfgang Becker (Becker-Hickl, Germany) * Paolo Bianchini (Università di Genova, Dipartimento di Fisica, Genova, I) * Ranieri Bizzarri (NEST-INFM, Scuola Normale Superiore, Pisa, I) * Rolf Th.Borlinghaus (Leica Microsystems CMS, Mannheim, Germany) * Fred Brakenhoff (University of Amsterdam, Swammerdam Institute for LIfe Sciences, NL) * Carlos Bustamante (UC Berkeley, Department of Molecular and Cellular Biology, CA, USA) * Valentina Caorsi (Università di Genova, Dipartimento di Fisica, Genova, I) * Giuseppe Chirico (Università di Milano-Bicocca, Dipartimento di Fisica, Milano, I) * Mario Faretta (IFOM-IEO Campus for Oncogenomics, European Institute of Oncology, Milano, I) * Stefan W. Hell (Max-Planck-Institute for Biophysical Chemistry, Dept. NanoBiophotonics, Goettingen, Germany) * Lucie Kubinova (Institute of Physiology, Academy of Sciences of the Czech Republic, Department of Biomathematics, Prague, Czech Republic) * Manuel Martinez Corral (University of Valencia, Department of Optics, ES) * Davide Mazza (Università di Genova, Dipartimento di Fisica, Genova, I) * Valentina Mussi (NANOMED, Università di Genova, I) * Erwin Neher (Max Planck Institute for Biophysical Chemistry, Goettingen, Germany) (to be confirmed) * Dario Parazzoli (IFOM-IEO Campus for Oncogenomics, European Institute of Oncology, Milano, I) * Francesco Pavone (LENS, Università di Firenze, I) * Tullio Pozzan (University of Padova, Department of Biomedical Sciences, Padova, I) * Franco Quercioli (Istituto dei Sistemi Complessi, CNR, Firenze, I) * Gimmi Ratto (Istituto di Neuroscienze, CNR, Pisa, I) * Peter Saggau (Baylor College of Medicine, Dept. Neuroscience, Houston, TX, USA) * Bruno Samorì (Università di Bologna, Dipartimento di Biochimica, Bologna, I) * Ilaria Testa (Università di Genova, Dipartimento di Fisica, Genova, I) * Giuseppe Vicidomini (Università di Genova, Dipartimento di Informatica e Scienze dell'Informazione, Genova, I) * Tony Wilson (University of Oxford, Department of Engineering Science, UK) * Fred Wouters (The Neuroscience Institute, European Neuroscience Institute, Goettingen, Germany)
MAIN SPONSOR: Leica Microsystems, Germany. SUPPORTING SPONSORS (to be updated): ISS, Urbana, Illinois; Becker and Hickl; OKO-LAB; Coherent; Springer and Verlag; Amici del Festival della Scienza.
REGISTRATION The number of students admitted is restricted to 30. The participation fee is 300 Euros which includes five nights accommodation and attendance at the lessons. The participation fee is reduced to 150 Euros in case the attendant does not need accommodation. The participation fee is reduced to 250 Euros for those registering by 15 December 2006. Applicants are admitted to the School by the Scientific Committee based on the information (short curriculum) provided on the pre-registration form, which must be submitted on line using the following application form. The admittance will be individually communicated via e-mail.
FURTHER DETAILS AND UPDATES http://sibpa.itc.it/ (section Biophysics School) http://www.lambs.it (section Events) ------------------------------------------------------------------------ ------------------------------------------------------------------------ ------------------------------------------------------------------------ ---------------------
--------------------------------------------- [...] Dance with wolves and count the stars, including the unseen...(L.Ferlinghetti, Challenges to young poet, 2001)
--------------------------------------------- Alberto Diaspro,LAMBS-IFOM MicroScoBIO Research Center, 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 ----------------------------------------------
==============================Original Headers============================== 24, 19 -- From diaspro-at-fisica.unige.it Fri Nov 17 09:51:34 2006 24, 19 -- Received: from phobos.ge.infm.it (phobos.ge.infm.it [193.205.153.2]) 24, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAHFpWw8005820; 24, 19 -- Fri, 17 Nov 2006 09:51:33 -0600 24, 19 -- Received: from [193.205.153.216] (albypc.ge.infm.it [193.205.153.216]) 24, 19 -- by phobos.ge.infm.it (8.11.6/8.11.0) with ESMTP id kAHGqnZ25981; 24, 19 -- Fri, 17 Nov 2006 17:52:49 +0100 24, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 24, 19 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 24, 19 -- Message-Id: {05E62BDA-B9A3-4D0D-9BBD-6CECE6D56512-at-fisica.unige.it} 24, 19 -- Cc: MicroscopyListSpamFilter-at-Microscopy.Com, microscopy-at-Microscopy.Com 24, 19 -- From: diaspro {diaspro-at-fisica.unige.it} 24, 19 -- Subject: School of Biophysics 2007 on Advanced Optical Methods - Venice 29 Jan-2 Feb 2007 24, 19 -- Date: Fri, 17 Nov 2006 16:51:30 +0100 24, 19 -- To: mplsm-users-at-yahoogroups.com, 24, 19 -- Confocal List Microscopy {CONFOCAL-at-LISTSERV.BUFFALO.EDU} 24, 19 -- X-Mailer: Apple Mail (2.752.2) 24, 19 -- Content-Transfer-Encoding: 8bit 24, 19 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAHFpWw8005820 ==============================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, energy dispersive x-ray analysis, electron energy loss spectroscopy, electron diffraction, electron tomography and ultramicrotomy preferred. A background in polymer science 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. Recent degree recipients as well as experienced practitioners are invited to apply for this position. The position is located in either Midland, Michigan or Freeport Texas. 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,
William A. Heeschen, Ph.D. Microscopy, Digital Imaging The Dow Chemical Company Midland, MI 48667 waheeschen-at-dow.com
==============================Original Headers============================== 8, 22 -- From WAHeeschen-at-dow.com Fri Nov 17 12:36:30 2006 8, 22 -- Received: from mail130.messagelabs.com (mail130.messagelabs.com [216.82.250.163]) 8, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAHIaU1u020117 8, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Nov 2006 12:36:30 -0600 8, 22 -- X-VirusChecked: Checked 8, 22 -- X-Env-Sender: WAHeeschen-at-dow.com 8, 22 -- X-Msg-Ref: server-15.tower-130.messagelabs.com!1163788588!8040606!1 8, 22 -- X-StarScan-Version: 5.5.10.7; banners=-,-,- 8, 22 -- X-Originating-IP: [216.99.65.28] 8, 22 -- Received: (qmail 15936 invoked from network); 17 Nov 2006 18:36:29 -0000 8, 22 -- Received: from mail8.dow.com (HELO mante89.nam.dow.com) (216.99.65.28) 8, 22 -- by server-15.tower-130.messagelabs.com with SMTP; 17 Nov 2006 18:36:29 -0000 8, 22 -- Received: by mante89.nam.dow.com with Internet Mail Service (5.5.2658.3) 8, 22 -- id {W5R7YRAB} ; Fri, 17 Nov 2006 13:36:28 -0500 8, 22 -- Message-ID: {0F0E6B7B4F6AE84CBC5E7AF57057FBA8084E8A-at-USMDLMDOWX022.dow.com} 8, 22 -- From: "Heeschen, Bill (WA)" {WAHeeschen-at-dow.com} 8, 22 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 8, 22 -- Subject: Job Posting from Dow Chemical for a microscopist 8, 22 -- Date: Fri, 17 Nov 2006 13:36:50 -0500 8, 22 -- MIME-Version: 1.0 8, 22 -- X-Mailer: Internet Mail Service (5.5.2658.3) 8, 22 -- Content-Type: text/plain ==============================End of - Headers==============================
The EH&S MSDS requesting system is primarily intended for the use of departments and affiliates of Iowa State University. EH&S reserves the right to refuse off-campus requests at their discretion, and will attempt to contact the requester via email if their request is refused.
***************
Many university resources block or limit access for security reasons. A European or Asian educational user may not have the same access as a North American educational user. I certainly cannot access the same resources from my home computer (a commercial internet service provider)that I can access from my university computer. --it's the price we pay due to misguided use of the internet.
-- 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============================== 13, 28 -- From Larry.Ackerman-at-ucsf.edu Fri Nov 17 12:45:36 2006 13, 28 -- Received: from emfmcb02.ucsfmedicalcenter.org (emfmcb02.ucsfmedicalcenter.org [64.54.46.98]) 13, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAHIjZ0A030779 13, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 17 Nov 2006 12:45:36 -0600 13, 28 -- Received: from 64.54.128.153 by emfmcb02.ucsfmedicalcenter.org with 13, 28 -- ESMTP (Tumbleweed Email Firewall SMTP Relay (Email Firewall v6.1.0)); 13, 28 -- Fri, 17 Nov 2006 10:57:48 -0800 13, 28 -- X-Server-Uuid: 44B63953-633F-4500-B2DA-A3E478718104 13, 28 -- Received: from [128.218.123.88] ([128.218.123.88]) by 13, 28 -- exvs06.net.ucsf.edu with Microsoft SMTPSVC(6.0.3790.1830); Fri, 17 Nov 13, 28 -- 2006 10:45:04 -0800 13, 28 -- Message-ID: {455E032F.5090204-at-ucsf.edu} 13, 28 -- Date: Fri, 17 Nov 2006 10:45:03 -0800 13, 28 -- From: "Larry Ackerman" {larry.ackerman-at-ucsf.edu} 13, 28 -- Reply-to: larry.ackerman-at-ucsf.edu 13, 28 -- Organization: UCSF, NeuroAnatomy 13, 28 -- User-Agent: Mozilla Thunderbird 1.0.7 (Macintosh/20050923) 13, 28 -- X-Accept-Language: en-us, en 13, 28 -- MIME-Version: 1.0 13, 28 -- To: Microscopy-at-microscopy.com 13, 28 -- Subject: MSDS corrections 13, 28 -- X-OriginalArrivalTime: 17 Nov 2006 18:45:05.0819 (UTC) 13, 28 -- FILETIME=[7FB3C6B0:01C70A78] 13, 28 -- X-WSS-ID: 6940D9A61GO31294-11-01 13, 28 -- Content-Type: text/plain; 13, 28 -- charset=iso-8859-1; 13, 28 -- format=flowed 13, 28 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
When exposed to fluorescent illumination, will all fluorescent objects in the field of view bleach at the same extent? ie. will brilliant objects fade less than dim objects?
Thank you, have a nice week-end!
Marie-Claude Bélanger Montréal
_________________________________________________________________ Achetez ce que vous voulez, quand vous voulez sur Sympatico / MSN Magasiner http://magasiner.sympatico.msn.ca/content/shp/?ctId=101,ptnrid=176,ptnrdata=081805
==============================Original Headers============================== 6, 21 -- From mcbelanger6-at-hotmail.com Fri Nov 17 13:02:32 2006 6, 21 -- Received: from bay0-omc3-s5.bay0.hotmail.com (bay0-omc3-s5.bay0.hotmail.com [65.54.246.205]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAHJ2Wsn009317 6, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 17 Nov 2006 13:02:32 -0600 6, 21 -- Received: from hotmail.com ([64.4.17.41]) by bay0-omc3-s5.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 21 -- Fri, 17 Nov 2006 11:02:31 -0800 6, 21 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 6, 21 -- Fri, 17 Nov 2006 11:02:31 -0800 6, 21 -- Message-ID: {BAY111-F31BD14A9D96DC5B8054BB7E3E80-at-phx.gbl} 6, 21 -- Received: from 64.4.17.200 by by111fd.bay111.hotmail.msn.com with HTTP; 6, 21 -- Fri, 17 Nov 2006 19:02:28 GMT 6, 21 -- X-Originating-IP: [66.46.83.20] 6, 21 -- X-Originating-Email: [mcbelanger6-at-hotmail.com] 6, 21 -- X-Sender: mcbelanger6-at-hotmail.com 6, 21 -- From: =?iso-8859-1?B?TWFyaWUtQ2xhdWRlIELpbGFuZ2Vy?= {mcbelanger6-at-hotmail.com} 6, 21 -- To: Microscopy-at-MSA.Microscopy.Com 6, 21 -- Subject: LM-Photobleaching 6, 21 -- Date: Fri, 17 Nov 2006 19:02:28 +0000 6, 21 -- Mime-Version: 1.0 6, 21 -- Content-Type: text/plain; charset=iso-8859-1; format=flowed 6, 21 -- X-OriginalArrivalTime: 17 Nov 2006 19:02:31.0424 (UTC) FILETIME=[EEEE7000:01C70A7A] ==============================End of - Headers==============================
Particle tracking is inbuilt into MetaMorph Premiere - applications Track Objects and Track Points are under Apps. With track points you manually click on the centre of the point (or the bit you are tracking. If the image is 'calibrated' under MetaMorph the App will give you micron distance travelled, velocity etc when logged to Excel, but to get a fix on the actual position you will need the X,Y co-ordinates from the data output (typically related to say the top left corner of the image and the pixel size - say 1024 x 1024). If you are just tracking within the image this is all you need (but you need to keep the pixel size of the images constant or calibrate/correct the image size). Time-lapse interval just needs to be set so that you can see which particle is which as they move.
If you are tracking beyond the image (using a motorised stage & perhaps raster scanning) then you need to figure in the motorised stage co-ordinates. To set the same 0,0 origin on a motorised stage I always used the same point on an England Finder slide (Graticules.com) but some use the X,Y stops which is probably not as accurate (I'll send my 'England Finder' help guide as well). Raster scans are set to the image size and so are easy to estimate distance across scans. For some odd reason our microscopes were bought without motorised stages [before my time], so I haven't used MetaMorph with a MetaMorph controlled motorised stage but I assume Metamorph can extrapolate the particles X,Y co-ordinates right back to this 0,0 origin on the slide (this may be beyond the Raster/snake scan and you may have to calculate relative to this start point). Track objects is similar etc track points except that this can use thresholding to locate the objects (not always successful so try everything like contrast enhancement (DIC/Phase), autofluorescence, dilution etc.. to get discreet particles). There are bugs in MetaMorph 6 (the version I use) regarding tracking co-ordinates so I'll email you my help pdf's and associated excel spreadsheet discussing this (later versions may have this bug fixed).
When manually clicking on a point in each image be careful of moving slightly back and forward across each subsequent image, as this will clock up a large distance travelled (back and forwards) even if the object hasn't moved at all - a time plot of all the movement X,Y points will demonstrate the object hasn't moved, and a calculation of the distance from the first point to last point will provide the actual distance moved - do this in Excel or MatLab. You may find that MatLab may be useful for calculations once you have the X,Y co-ordinates, but generally I have always got MetaMorph to do everything I need provided I spend a while thinking about it (some aspects of the software are rather poor - like no manual binary image editor for thresholded images - but it always gets the job done and it knocks the spots off simpler ImageJ (for a price) and MatLab (for ease of use and inbuilt image analysis routines). Do email MetaMorph support if you get into problems (or you local rep who will generally be very helpful).
Unfortunate my 5-year contract with the Institute of Opthalmology has just terminated (I'm 'between' jobs at the moment) - so I have lost access to my MetaMorph key, but hopefully my help guides will get you going (note that they are only written with screenshots to remind me what to do, they aren't proof read). Likewise my UCL email address will no doubt fail soon.
If you don't have the Apps (they may not be available under MetaMorph Basic) just use XY co-ordinates from Metamorph object threshold/measurements or perhaps manual count (I think that gives the X,Y co-ordinates if logged to Excel).
Hope this helps.
Keith
MetaMorph http://www.moleculardevices.com
--------------------------------------------
Dr Keith J Morris Manager Imaging Facilties Cell Biology Division The institute of Ophthalmology 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {sharad-at-post.harvard.edu} To: {keith.morris-at-ucl.ac.uk} Sent: Friday, November 17, 2006 3:30 AM
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Title-Subject: [Filtered] measurement of fluorescence
Question: Occasionally the question of measuring a fluorescence signal inside a yeast cell arises (specifically gfp tagged protein localizing to an organelle). We have a Zeiss Axioskop and use the Axiovision software. Our axiocam takes b/w digital images. The aim to measure at least a hundred cells each from various mutants. Anyone have suggestions or advice?
Western Michigan University, a leading institution of higher learning (Carnegie Doctoral/Research Extensive) in one of the nation’s most enterprising regions, seeks an internationally recognized scholar with demonstrated administrative accomplishment and international scholarly achievement to head the Diether H. Haenicke Institute for Global Education. The person who is chosen will have outstanding credentials and experience for leading the Haenicke Institute in globalizing the University, enhancing international partnerships, administering selected academic programs and centers, supervising and supporting international students, organizing study abroad for WMU students, and working with faculty and staff in promoting relevant scholarly and creative inquiry. We seek a person who has substantial international experience and who is committed to obtain support and external funding for international programs and scholarship. Our choice shall be able to speak and work in more than one language and shall have extensive experience residing and working or studying outside the United States. Founded in 1998, the Institute is named in honor of the distinguished humanist and WMU President Diether H. Haenicke. Detailed information, including a complete description of the position, may be found at *http://international.wmich.edu* {http://international.wmich.edu/} . The head of the Institute shall have the title of Dean and shall report to the Provost and Vice President for Academic Affairs. The Haenicke Dean shall possess the credentials to be named full professor at the University.
Applications or nominations for this position should consist of a letter of interest or a description of qualifications, along with a curriculum vitae and contact information for five references who may be approached for a recommendation, addressed to: Dr Lewis Pyenson, Chair of the Search Committee, Dean of the Graduate College, Western Michigan University, Kalamazoo, MI 49008-5242, *hige-deansearch-at-wmich.edu {mailto:hige-deansearch-at-wmich.edu} *. For full consideration, applications and supporting documentation should be received by 15 December 2006.
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--
Butterflies "can fly because they take themselves lightly"
Nancy Smythe wrote: ==================================== Question: We also had the problem several months ago and not just in my lab but the core facility at the university alse. We found, after much frustration, that the older the UA the better it works. Must be something in the processing of the newer batches that is causing this. Lucky for me I found some in the back of the cabinet that will keep me going for a long time. ==================================== The uranyl acetate used in EM laboratories, does not all come from the same place. The problems that were experienced seem to have been traced to a particular source.
SPI Supplies, and presumably other suppliers, presently have in stock "fresh" product, it is very fast dissolving, and works just fine in the hands of customers who have purchased and used it. I don't think it is correct to say that "older is better" (unless you are comparing some of the "problem" lot material with older but good material.
If someone has had problems with the SPI-Chem brand of uranyl acetate from SPI Supplies, I would like to know about it, especially if such problems have been relatively recent. So far as I know, our "fresh" UA works just fine.
Disclaimer: SPI Supplies is a supplier of uranyl acetate for EM laboratories.
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
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 10, 26 -- From cgarber-at-2spi.com Mon Nov 20 02:18:51 2006 10, 26 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAK8Ipe5010961 10, 26 -- for {microscopy-at-msa.microscopy.com} ; Mon, 20 Nov 2006 02:18:51 -0600 10, 26 -- Received: from yourb27fb1c401 (c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118]) 10, 26 -- (authenticated bits=0) 10, 26 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id kAK8Ii59021853; 10, 26 -- Mon, 20 Nov 2006 03:18:50 -0500 10, 26 -- X-IDV-FirstRcvd: c-71-230-36-118.hsd1.pa.comcast.net [71.230.36.118] 10, 26 -- X-IDV-HELO: yourb27fb1c401 10, 26 -- X-IDV-Authenticated-User: cgarber 10, 26 -- Message-ID: {01a601c70c7c$7f4c3fd0$6401a8c0-at-yourb27fb1c401} 10, 26 -- From: "Garber, Charles A" {cgarber-at-2spi.com} 10, 26 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 10, 26 -- Subject: Uranyl acetate 10, 26 -- Date: Mon, 20 Nov 2006 03:18:43 -0500 10, 26 -- MIME-Version: 1.0 10, 26 -- Content-Type: text/plain; 10, 26 -- format=flowed; 10, 26 -- charset="iso-8859-1"; 10, 26 -- reply-type=original 10, 26 -- Content-Transfer-Encoding: 7bit 10, 26 -- X-Priority: 3 10, 26 -- X-MSMail-Priority: Normal 10, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 10, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
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A question for those who have an Iridium sputter coater for HR-SEM.
What are your usual sputter conditions for high resolution : target diameter, sample to target distance, current, voltage, gas pressure, runtime ?
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
We are in need of a sample holder, single or double tilt, to fit a Philips/FEI EM-420. If anyone has one they are no longer using contact me offline. Thanks, David
****************************** David C. Garrett, Ph.D. Supervisor, Electron Microscopy Dept. Materials Science University of North Texas P.O. Box 305310 Denton, TX 76203-5310 940.565.3964 dgarrett-at-unt.edu ******************************
==============================Original Headers============================== 3, 27 -- From dgarrett-at-unt.edu Mon Nov 20 10:13:18 2006 3, 27 -- Received: from mailhost.unt.edu (mailhost.unt.edu [129.120.188.67]) 3, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAKGDIXu026134 3, 27 -- for {microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 10:13:18 -0600 3, 27 -- Received: from iatro (localhost.localdomain [127.0.0.1]) 3, 27 -- by mail5 (Postfix) with SMTP id 31CD237B87 3, 27 -- for {microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 10:12:47 -0600 (CST) 3, 27 -- X-Spam-Checker-Version: SpamAssassin 3.1.7 (2006-10-05) on 3, 27 -- mail5.ncsmail.private.unt.edu 3, 27 -- X-Spam-Level: 3, 27 -- X-Spam-Status: No, score=-6.0 required=6.0 tests=UNT_LOCAL_FROM 3, 27 -- autolearn=disabled version=3.1.7 3, 27 -- Received: from GWIA.unt.edu (gwia.unt.edu [129.120.221.21]) 3, 27 -- by mailhost.unt.edu (Postfix) with ESMTP id EC9D737BD7 3, 27 -- for {microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 10:12:37 -0600 (CST) 3, 27 -- Received: from SMTP-MTA by GWIA.unt.edu 3, 27 -- with Novell_GroupWise; Mon, 20 Nov 2006 10:13:08 -0600 3, 27 -- Message-Id: {45617FAA020000CC0000AB9C-at-GWIA.unt.edu} 3, 27 -- X-Mailer: Novell GroupWise Internet Agent 7.0.1 3, 27 -- Date: Mon, 20 Nov 2006 10:12:58 -0600 3, 27 -- From: "David Garrett" {dgarrett-at-unt.edu} 3, 27 -- To: {microscopy-at-microscopy.com} 3, 27 -- Subject: Need: sample holder Philips/FEI 400 series 3, 27 -- Mime-Version: 1.0 3, 27 -- Content-Type: text/plain; charset=US-ASCII 3, 27 -- Content-Transfer-Encoding: 7bit 3, 27 -- Content-Disposition: inline ==============================End of - Headers==============================
Using a Denton Desk IV TSC, the target is 55mm diameter and distance to SEM stub is about 20mm.
Typical conditions are 15mT and 15mA (unknown voltage). Depending on rotation %, 45 seconds works well. Faster rotation seems to require longer time.
gary g.
At 07:45 AM 11/20/2006, you wrote:
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==============================Original Headers============================== 11, 22 -- From gary-at-gaugler.com Mon Nov 20 10:31:10 2006 11, 22 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAKGV98d004776 11, 22 -- for {microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 10:31:10 -0600 11, 22 -- Received: (qmail 8285 invoked from network); 20 Nov 2006 08:31:09 -0800 11, 22 -- Received: by simscan 1.1.0 ppid: 8252, pid: 8283, t: 0.1647s 11, 22 -- scanners: regex: 1.1.0 attach: 1.1.0 11, 22 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 11, 22 -- by qsmtp3 with SMTP; 20 Nov 2006 08:31:09 -0800 11, 22 -- Message-Id: {7.0.1.0.2.20061120082146.024e0448-at-gaugler.com} 11, 22 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 11, 22 -- Date: Mon, 20 Nov 2006 08:31:02 -0800 11, 22 -- To: jacques.faerber-at-ipcms.u-strasbg.fr 11, 22 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 22 -- Subject: Re: [Microscopy] SEM Ir coating quest. 11, 22 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 22 -- In-Reply-To: {200611201545.kAKFjXDP017585-at-ns.microscopy.com} 11, 22 -- References: {200611201545.kAKFjXDP017585-at-ns.microscopy.com} 11, 22 -- Mime-Version: 1.0 11, 22 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 11, 22 -- Content-Transfer-Encoding: 8bit 11, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAKGV98d004776 ==============================End of - Headers==============================
I'm making up a 5% ammonium molybdate / 1% trehalose w/v negative stain for a Special Project. Is there any trick to this? The solution is staying stubbornly milky so far. Does this clear up after doing a pH adjustment or is it just hard to get into solution?
Thanks!
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 6, 23 -- From TindallR-at-missouri.edu Mon Nov 20 14:47:28 2006 6, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAKKlRTs022628 6, 23 -- for {microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 14:47:28 -0600 6, 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); 6, 23 -- Mon, 20 Nov 2006 14:47:27 -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: Neg stain: ammonium molybdate 6, 23 -- Date: Mon, 20 Nov 2006 14:47:27 -0600 6, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68BDA-at-UM-XMAIL08.um.umsystem.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: Neg stain: ammonium molybdate 6, 23 -- Thread-Index: AccM5RWgDOs3TqeNTwO4ZsYvy8qGow== 6, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 6, 23 -- To: {microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 20 Nov 2006 20:47:27.0450 (UTC) FILETIME=[16E503A0:01C70CE5] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAKKlRTs022628 ==============================End of - Headers==============================
Why 5% AM. Way back when, I routinely used a 1% AM. If I remember correctly it did take a little while for the AM to dissolve, maybe overnight????
The sugar may also interfere with the dissolving of the AM, but I have no idea about that aspect.
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: Monday, November 20, 2006 3:53 PM To: Calomeni, Edward
Dear Listers,
I'm making up a 5% ammonium molybdate / 1% trehalose w/v negative stain for a Special Project. Is there any trick to this? The solution is staying stubbornly milky so far. Does this clear up after doing a pH adjustment or is it just hard to get into solution?
Thanks!
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 6, 23 -- From TindallR-at-missouri.edu Mon Nov 20 14:47:28 2006 6, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAKKlRTs022628 6, 23 -- for {microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 14:47:28 -0600 6, 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); 6, 23 -- Mon, 20 Nov 2006 14:47:27 -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: Neg stain: ammonium molybdate 6, 23 -- Date: Mon, 20 Nov 2006 14:47:27 -0600 6, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68BDA-at-UM-XMAIL08.um.umsystem.edu} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: Neg stain: ammonium molybdate 6, 23 -- Thread-Index: AccM5RWgDOs3TqeNTwO4ZsYvy8qGow== 6, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 6, 23 -- To: {microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 20 Nov 2006 20:47:27.0450 (UTC) FILETIME=[16E503A0:01C70CE5] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAKKlRTs022628 ==============================End of - Headers==============================
==============================Original Headers============================== 18, 31 -- From Edward.Calomeni-at-osumc.edu Mon Nov 20 16:28:05 2006 18, 31 -- Received: from pluto.osumc.edu (pluto.osumc.edu [140.254.120.27]) 18, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAKMS47a003233 18, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 16:28:04 -0600 18, 31 -- Received: from localhost (unknown [127.0.0.1]) 18, 31 -- by pfeg01.osumc.edu (Postfix) with ESMTP id ABA5F14103; 18, 31 -- Mon, 20 Nov 2006 17:28:01 -0500 (EST) 18, 31 -- Received: from pfeg01.osumc.edu ([127.0.0.1]) 18, 31 -- by localhost (pfeg01.osumc.edu [127.0.0.1]) (amavisd-new, port 10024) 18, 31 -- with LMTP id 23883-01-66; Mon, 20 Nov 2006 17:28:01 -0500 (EST) 18, 31 -- Received: from msxc01.OSUMC.EDU (msxc01.osumc.edu [10.127.29.33]) 18, 31 -- by pfeg01.osumc.edu (Postfix) with ESMTP id 86C01140F7; 18, 31 -- Mon, 20 Nov 2006 17:28:01 -0500 (EST) 18, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 18, 31 -- Content-class: urn:content-classes:message 18, 31 -- MIME-Version: 1.0 18, 31 -- Content-Type: text/plain; 18, 31 -- charset="us-ascii" 18, 31 -- Subject: RE: [Microscopy] Neg stain: ammonium molybdate 18, 31 -- Date: Mon, 20 Nov 2006 17:28:01 -0500 18, 31 -- Message-ID: {7A541592F9A53A4E86E7A3D5C4C7F68CB4A9BC-at-msxc01.OSUMC.EDU} 18, 31 -- In-Reply-To: {200611202052.kAKKqZxi030051-at-ns.microscopy.com} 18, 31 -- X-MS-Has-Attach: 18, 31 -- X-MS-TNEF-Correlator: 18, 31 -- Thread-Topic: [Microscopy] Neg stain: ammonium molybdate 18, 31 -- Thread-Index: AccM5jQ5erKdqArMRT6NZ/2KK1EGCwADHCHg 18, 31 -- From: "Calomeni, Edward " {Edward.Calomeni-at-osumc.edu} 18, 31 -- To: {TindallR-at-missouri.edu} , {Microscopy-at-microscopy.com} 18, 31 -- X-Virus-Scanned: by amavisd-new at osumc.edu 18, 31 -- Content-Transfer-Encoding: 8bit 18, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAKMS47a003233 ==============================End of - Headers==============================
I've never had a problem with molybdate. The milkiness suggests two things, pH or concentration. Several thoughts which may help.
First, what is the pH you are adjusting to. Ammonium molubdate is naturally weakly acidic. I can adjust the pH to 6.0 readily, but I have found high pH (} 6.0) leads to poor solubility.
Second, the concentration I use is 8.75mM, or just over 1%. This is adjusted so that the number of dense atoms is controled between different stains - eg. the number of Mo atoms able to deflect the beam at 8.75mM is the same as the number of W in 2.5mM PTA or U in any formulation of 60mM uranyl stain (acetate, oxylate, formate, sulfate, etc) You are using 5% - pretty high. Because the stain will dry down, do you really need that high of a concentration.
Third. Never used trehalose, but been tempted to give it a try. Does 1% trehalose go into solution readily. Does it effect the pH of the solution - perhaps raising it above 7.0? Will trehalose go into solution at lower pH? Can you adjust the pH to between 3.0 and 6.0, and if you do, what happens?
Finally, when you mix the two compounds you will effect the saturation point for the two, and lower the relative solubility of each component. Solubility of AmMo is quite high. I don't know the solubility of trehalose, but Harris has used 10% solutions, and it's insoluble in organic solvents, and weakly soluble in alcohol. Perhaps you are affecting the saturation point of the trehalose by mixing with AmMo. I know Harris has done work as you describe, and believe Charles Humphrey at the CDC may have also, but I do not know if there are any side issues.
I have tried mixed stains. Many times they reacted and caused all sorts of precipitates, colloidal milkiness, etc. Maybe they just won't work together under the conditions you are using.
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============================== 14, 20 -- From paul_hazelton-at-umanitoba.ca Mon Nov 20 16:36:46 2006 14, 20 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAKMajXK013864 14, 20 -- for {microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 16:36:45 -0600 14, 20 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 14, 20 -- (authenticated bits=0) 14, 20 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id kAKMZM6N005739; 14, 20 -- Mon, 20 Nov 2006 16:35:23 -0600 (CST) 14, 20 -- Message-ID: {45622DA3.8080408-at-umanitoba.ca} 14, 20 -- Date: Mon, 20 Nov 2006 16:35:15 -0600 14, 20 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 14, 20 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 14, 20 -- X-Accept-Language: en-us, en 14, 20 -- MIME-Version: 1.0 14, 20 -- To: TindallR-at-missouri.edu, Microscopy Listserver {microscopy-at-microscopy.com} 14, 20 -- Subject: Re: [Microscopy] Neg stain: ammonium molybdate 14, 20 -- References: {200611202050.kAKKo09W026162-at-ns.microscopy.com} 14, 20 -- In-Reply-To: {200611202050.kAKKo09W026162-at-ns.microscopy.com} 14, 20 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 14, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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Email: jensim-at-inbox.com Name: J S
Title-Subject: [Filtered] Kalling's No 2 for duplex stainless steel.
Question: May you explain me the microstructure revealed in duplex stainless steel etched with Kalling's No2? Which phase is the ferrite and which the austenite? Thank you in advance for your time. I am looking forward to receiving a reply from you
This Question was submitted to Ask-A-Microscopist by (jcraft-at-memphis.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, November 20, 2006 at 10:55:06 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both jcraft-at-memphis.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: jcraft-at-memphis.edu Name: jackie williams
I'm puzzled by the interpretation and value of EBIC as specimen current monitor versus simple SE detection.
If a specimen is imaged with E-T SE and then with specimen current monitor (SCM) via stage current monitor, what might the differences be? I see them as a measure of conductivity for SCM whereas the SE is generation of reflected SEs.
Does the nature of reflected SEs directly correlate to the amount of SCM?
What info can be gleaned by using SCM versus SE?
All thoughts invited.
gary g.
==============================Original Headers============================== 8, 17 -- From gary-at-gaugler.com Mon Nov 20 20:32:59 2006 8, 17 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 8, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAL2WwcS018371 8, 17 -- for {microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 20:32:58 -0600 8, 17 -- Received: (qmail 2638 invoked from network); 20 Nov 2006 18:32:58 -0800 8, 17 -- Received: by simscan 1.1.0 ppid: 2617, pid: 2636, t: 0.0983s 8, 17 -- scanners: regex: 1.1.0 attach: 1.1.0 8, 17 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 8, 17 -- by qsmtp2 with SMTP; 20 Nov 2006 18:32:58 -0800 8, 17 -- Message-Id: {7.0.1.0.2.20061120182645.024cef88-at-gaugler.com} 8, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 8, 17 -- Date: Mon, 20 Nov 2006 18:32:51 -0800 8, 17 -- To: MSA listserver {microscopy-at-microscopy.com} 8, 17 -- From: Gary Gaugler {gary-at-gaugler.com} 8, 17 -- Subject: EBIC vs. SE 8, 17 -- Mime-Version: 1.0 8, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I need to prepare the LED chip for back side SIMS anaysis. SIMS depth profile resolution is significantly degraded if a large depth must be sputtered prior to the depth of analyticall interest. This problem can be exacerbated on LED chips which is covered thick electrode. When the LED chip sputtering which has thick eletrode exhibit strong nonuniform sputtering properties . These problems can be avoided if sufficient sample substrate material can be removed with adequate precision to allow SIMS depth profiling from the sample back side.
Does anyone have experience or suggestions on how to prepare back side SIMS samples? Method, Recipe for polishing step and so on..
Thanks in advance
Young Woo Jeong
==============================Original Headers============================== 10, 22 -- From jyw-at-lge.com Mon Nov 20 20:54:28 2006 10, 22 -- Received: from mail1.lge.co.kr (mail1.lge.co.kr [156.147.1.151]) 10, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAL2sR55029263 10, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 20:54:28 -0600 10, 22 -- Received: from [156.147.51.54] (jyw-at-lge.com) by 10, 22 -- mail1.lge.co.kr (Terrace MailWatcher) 10, 22 -- with ESMTP id 2006112111:54:22:863195.12230.155 10, 22 -- for {Microscopy-at-microscopy.com} ; 10, 22 -- Tue, 21 Nov 2006 11:54:22 +0900 (KST) 10, 22 -- Subject: TEM sample preparation for back side SIMS analysis 10, 22 -- Reply-To: "=?EUC-KR?B?waS/tb/s?=" {jyw-at-lge.com} 10, 22 -- From: "=?EUC-KR?B?waS/tb/s?=" {jyw-at-lge.com} 10, 22 -- To: Microscopy-at-microscopy.com 10, 22 -- Message-ID: {OF8DE4D82F.1F7B65DD-ON4925722D.000FF6CD-4925722D.000FF6CF-at-lge.com} 10, 22 -- Date: Tue, 21 Nov 2006 11:54:22 +0900 10, 22 -- Return-Receipt-To: "=?EUC-KR?B?waS/tb/s?=" {jyw-at-lge.com} 10, 22 -- X-MIMETrack: Serialize by Router on LGEKRHQMS03/LGE/LG Group(Release 6.5.4FP2HF559 | September 10, 22 -- 7, 2006) at 2006/11/21 11:54:22 10, 22 -- MIME-Version: 1.0 10, 22 -- Content-type: text/plain; charset=US-ASCII 10, 22 -- X-TERRACE-SPAMMARK: NOT spam-marked. 10, 22 -- (by Terrace) ==============================End of - Headers==============================
EBIC and specimen current are not the same thing. For specimen current imaging, you image the balance between the incident beam current (Ib), the current due to backscatters (Ibse), the current due to secondaries (Ise) and the specimen current (Isc), such that:
Ib=Ibse+Ise+Isc
Or, Isc = Ib - Ibse - Ise
Thus, specimen current is like looking at the opposite of the backscatter and secondary signals.
EBIC, on the other hand, is a semiconductor-specific technique. In specimen current, you put the current amplifier between the sample and ground. In EBIC, however, you put the current amplifier between the p- and n- sides of an electrically active diode. When beam electrons hit the sample, they can excite it in a way similar to light falling on a solar cell. The energy imparted by the beam causes current to flow through the external amplifier. EBIC then lets you image areas of high or low electron-hole-pair recombination activity in the imaged device. If you ground the n-side and operate above the unity yield point, you'll suppress most of the specimen current contribution and therefore most or all of the topography and get a purely semiconductor-device-physics signal.
EBIC signals will often be a few orders of magnitude higher than the beam current, whereas specimen currents tend to be on the same order as the beam current.
Chad Parish
==============================Original Headers============================== 8, 24 -- From chad.parish-at-gmail.com Tue Nov 21 06:15:44 2006 8, 24 -- Received: from wx-out-0506.google.com (wx-out-0506.google.com [66.249.82.232]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kALCFhDc018992 8, 24 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 06:15:43 -0600 8, 24 -- Received: by wx-out-0506.google.com with SMTP id h30so2392523wxd 8, 24 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 04:15:43 -0800 (PST) 8, 24 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 24 -- s=beta; d=gmail.com; 8, 24 -- h=received:message-id:date:from:to:subject:in-reply-to:mime-version:content-type:content-transfer-encoding:content-disposition:references; 8, 24 -- b=PhGzvNQ7UC97xmlnOuDy2I4ZtWOK7ralwb1CWANlAx7R7l4kvWqZsSJ71WhBVP7mDaXtIlVgsIxqCeSsTvrw+OpmJ0ORQCN+3vIOWlb8jbBJdm9ox0CIHAi3vBmSAPsVIPuU+mnlxJ8HZhs3zez4ZQ1Y8ND+kFMVxUWuzXdYoaM= 8, 24 -- Received: by 10.90.51.17 with SMTP id y17mr5096615agy.1164111343406; 8, 24 -- Tue, 21 Nov 2006 04:15:43 -0800 (PST) 8, 24 -- Received: by 10.35.93.9 with HTTP; Tue, 21 Nov 2006 04:15:42 -0800 (PST) 8, 24 -- Message-ID: {fd59a7d40611210415u5fac01c3m87d4af1b567b2cba-at-mail.gmail.com} 8, 24 -- Date: Tue, 21 Nov 2006 07:15:42 -0500 8, 24 -- From: "Chad Parish" {chad.parish-at-gmail.com} 8, 24 -- To: microscopy-at-microscopy.com 8, 24 -- Subject: Re: [Microscopy] EBIC vs. SE 8, 24 -- In-Reply-To: {200611210240.kAL2eRDe028476-at-ns.microscopy.com} 8, 24 -- MIME-Version: 1.0 8, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 24 -- Content-Transfer-Encoding: 7bit 8, 24 -- Content-Disposition: inline 8, 24 -- References: {200611210240.kAL2eRDe028476-at-ns.microscopy.com} ==============================End of - Headers==============================
Being somewhat experienced in TEM, I am starting to work on a SEM (Tescan). The SEM is equipped with rotatory pump and TMP. There are 3 detectors: SE, BSE and EDX. In the documentation (VEGA software) it is written that the microscope can work in 2 modes: low vacuum or high vacuum. The high vacuum is allowed under a vacuum of 5x10-3 Pa. When I open the manual valve (to allow connections between the column and the chamber, thus to work in high-vacuum mode) and start pumping, the vacuum reaches no better than 8x10-3 Pa. In this condition I can only work in low vacuum mode. Nothing seems to go wrong, I wonder why I cannot reach higher vacuum?? Also, can get good results with the EDX detector in low-vacuum mode (well it is still 8x10-3 Pa)?
Stephane
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==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Tue Nov 21 06:22:37 2006 6, 18 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.91.144]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kALCMbth025087 6, 18 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 06:22:37 -0600 6, 18 -- Received: (qmail 9197 invoked by uid 60001); 21 Nov 2006 12:22:36 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.com; 6, 18 -- h=Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 6, 18 -- b=W23HU9Dsq7JxrlCsa5+iOP+g9giBMBtLEH6CcWhPl5Z3uxG/LrUnQbfLP1Eva5kyblg52vIL4UpngrJGhzFHohPVFkEWjSrsiS/+hkPCJkEoX0KI/QsQOvzXvAGMf2uzhpMzjbkxdOfsgcksJPEw8vrZy9+f+I2hC1DvMwpcZB4=; 6, 18 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Tue, 21 Nov 2006 04:22:36 PST 6, 18 -- Date: Tue, 21 Nov 2006 04:22:36 -0800 (PST) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: Basic SEM questions 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 6, 18 -- Message-ID: {609645.8917.qm-at-web37412.mail.mud.yahoo.com} ==============================End of - Headers==============================
Being somewhat experienced in TEM, I am starting to work on a SEM (Tescan). The SEM is equipped with rotatory pump and TMP. There are 3 detectors: SE, BSE and EDX. In the documentation (VEGA software) it is written that the microscope can work in 2 modes: low vacuum or high vacuum. The high vacuum is allowed under a vacuum of 5x10-3 Pa. When I open the manual valve (to allow connections between the column and the chamber, thus to work in high-vacuum mode) and start pumping, the vacuum reaches no better than 8x10-3 Pa. In this condition I can only work in low vacuum mode. Nothing seems to go wrong, I wonder why I cannot reach higher vacuum?? Also, can get good results with the EDX detector in low-vacuum mode (well it is still 8x10-3 Pa)?
Stephane
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==============================Original Headers============================== 6, 19 -- From nizets2-at-yahoo.com Tue Nov 21 06:22:40 2006 6, 19 -- Received: from web37411.mail.mud.yahoo.com (web37411.mail.mud.yahoo.com [209.191.91.143]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kALCMcBb025125 6, 19 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 06:22:39 -0600 6, 19 -- Received: (qmail 39750 invoked by uid 60001); 21 Nov 2006 12:22:38 -0000 6, 19 -- Message-ID: {20061121122238.39748.qmail-at-web37411.mail.mud.yahoo.com} 6, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 19 -- s=s1024; d=yahoo.com; 6, 19 -- h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 6, 19 -- b=cfOvs4tchgPpQxEOAN2L8O38oH6JekU8ads8Smu5P1rdhw1aFEV78kZAXOpG1QlKCVJk4XSAZUczFHJVIsVi1Ui5zWWWlzuZ6kQPUdclIDOCVmV3ISvdVDJAW+fIREx4VC9botY8xyVxTNwKD9IlYXwxx3aHheu5hgTxuR4kKnk=; 6, 19 -- X-YMail-OSG: zxBn1kQVM1khwB2gYlNRp4xq0_aTuBa2ux8wx52BmaHO7PCUqyb8GuspW8MypmwodXaDY3nV7wM81XghDrZCNHl96_E5kC9A15mPeBrhKnv1zQncRXZTuWa22qgiTjhH5AwB_KlBA.CoM_k- 6, 19 -- Received: from [80.122.101.102] by web37411.mail.mud.yahoo.com via HTTP; Tue, 21 Nov 2006 04:22:38 PST 6, 19 -- Date: Tue, 21 Nov 2006 04:22:38 -0800 (PST) 6, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 19 -- Subject: Basic SEM questions 6, 19 -- To: microscopy-at-microscopy.com 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==============================
My main advice would be: be careful with quantification of fluorescence. And also arm yourself with a lot of patience. If you consider that the amount of fluorescence will vary with the focus, you will have to focus each cell for the maximum fluorescence for a given organelle (don't measure several cells in one field). Don't use automatic exposition time, otherwise you cannot compare your results. Choose carefully one and only one exposition time and deal with it for ALL your cells. To avoid bias due to bleaching, you should determine a time period during which you make your adjustments (focus and field) before taking the picture and use the same time for ALL your cells (I think that photobleaching is an important factor with GPF constructs). Be careful not to choose a cell which was already exposed (in the field of view of another cell of interest). Now if you have an option to automatize the task, I guess these biases would be averaged by measuring several hundreds of cells (though bleaching will still be an issue, but you can control it).
Good luck Stephane
--- moakes-at-HCRcenter.com wrote:
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==============================Original Headers============================== 12, 20 -- From nizets2-at-yahoo.com Tue Nov 21 06:26:52 2006 12, 20 -- Received: from web37411.mail.mud.yahoo.com (web37411.mail.mud.yahoo.com [209.191.91.143]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kALCQq5p006396 12, 20 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 06:26:52 -0600 12, 20 -- Received: (qmail 40682 invoked by uid 60001); 21 Nov 2006 12:26:51 -0000 12, 20 -- Message-ID: {20061121122651.40680.qmail-at-web37411.mail.mud.yahoo.com} 12, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 12, 20 -- s=s1024; d=yahoo.com; 12, 20 -- h=Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 12, 20 -- b=jLmF+79C7Rtea/6OjxjoumspTTzcIZ8DrQDhnQJomgduKgiC3NSFO1Ev7l28gt7kLgUn7cAky0fVTFPaYXY5QtNk6kopiqGOVknAz0Aga27S+SK9iBG6EjrHBV2/MWRmLQbs87wgoPZyhmrUW1y36ywhj2xEDVTV1hKkmuK/XcE=; 12, 20 -- Received: from [80.122.101.102] by web37411.mail.mud.yahoo.com via HTTP; Tue, 21 Nov 2006 04:26:51 PST 12, 20 -- Date: Tue, 21 Nov 2006 04:26:51 -0800 (PST) 12, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 12, 20 -- Subject: Re: [Microscopy] viaWWW: measurement of fluorescence 12, 20 -- To: moakes-at-HCRcenter.com 12, 20 -- Cc: microscopy-at-microscopy.com 12, 20 -- In-Reply-To: {200611181439.kAIEdBUC027262-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==============================
I do not known the procedures on your microscope but others with a similar format would show this type of problem under the following conditions.
1. A genuine leak in the specimen chamber 2. The control valve for the VP (low vacuum) operation is not fully closed. 3. The valve between the VP pumping system and the chamber is not fully closed thus you are pumping against the second rotary pump.
Hope this may help?
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: {nizets2-at-yahoo.com} To: {protrain-at-emcourses.com} Sent: Tuesday, November 21, 2006 12:23 PM
X-from: "Dr. Michael Horst" {HORST_MN-at-Mercer.edu} To: "Greg Erdos" {gwe-at-ufl.edu} Sent: Tuesday, November 21, 2006 9:00 AM
Thanks to all who responded to my query about ammonium molybdate not wanting to dissolve. A couple people had questions about why I was using 5% AM, rather than 1-2%. The reason is simply that I was going by a recent Microscopy and Analysis May, 2006 article on developments in neg staining as my starting point. I may try a lesser concentration. It's experiment time.
Everything finally went into solution, by the way, especially after I adjusted the pH to 7.4 with NaOH.
I also tried 0.5% UA with 12mM oxalic acid, which supposedly allows pH adjustment into the physiological range. All was well until I got near ph 7.3, when it looked like all the UA precipitated out, leaving a nearly clear solution behind. I'm gonna try it anyway. You just never know.
Thanks again. If anyone's interested, I'll keep them posted on my negative (staining) results.
Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
==============================Original Headers============================== 7, 23 -- From TindallR-at-missouri.edu Tue Nov 21 08:50:37 2006 7, 23 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kALEobiI020778 7, 23 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 08:50:37 -0600 7, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 7, 23 -- Tue, 21 Nov 2006 08:50:36 -0600 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: Negative staining: Thanks 7, 23 -- Date: Tue, 21 Nov 2006 08:50:36 -0600 7, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68BDC-at-UM-XMAIL08.um.umsystem.edu} 7, 23 -- X-MS-Has-Attach: 7, 23 -- X-MS-TNEF-Correlator: 7, 23 -- Thread-Topic: Negative staining: Thanks 7, 23 -- Thread-Index: AccNfGd/rQsnGPh5R9Kmljbz31d4dA== 7, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 7, 23 -- To: {microscopy-at-microscopy.com} 7, 23 -- X-OriginalArrivalTime: 21 Nov 2006 14:50:36.0947 (UTC) FILETIME=[67A74630:01C70D7C] 7, 23 -- Content-Transfer-Encoding: 8bit 7, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kALEobiI020778 ==============================End of - Headers==============================
Hi Jackie I regularly work with formvar/carbon coated slot grids with over 100 serial sections on them. I feel your pain. I have done two things; 1) be very careful - you can use grid sticks or tweezers, but you have to be very careful 2) change your embedding techniques so that you do not have to stain on the grid.
I know that #1 is not very useful :-) I have developed an embedding technique that does not require staining on the grid. Both UA and Pb are done in-block. The 50nm sections have fine contrast. This means that I do not have to stain the grids and risk the sections or the formvar. If you want the protocol, let me know. David
_____________________
David Elliott Ph.D. Assistant Professor - Department of Cell Biology and Anatomy Director, Research Microscopy Core Service University of Arizona College of Medicine PO Box 245004 Tucson, AZ 85724
Voice: 520-626-7870 Fax: 520-626-2097
On Nov 20, 2006, at 5:17 PM, jcraft-at-memphis.edu wrote:
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==============================Original Headers============================== 12, 22 -- From Elliott-at-arizona.edu Tue Nov 21 09:39:23 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 kALFdMVu032200 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Nov 2006 09:39:22 -0600 12, 22 -- Received: from localhost (amavis5.email.arizona.edu [10.0.0.208]) 12, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP 12, 22 -- id 7F296121CC52; Tue, 21 Nov 2006 08:39:22 -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 12, 22 -- id 00767121C1FD; Tue, 21 Nov 2006 08:39:20 -0700 (MST) 12, 22 -- Mime-Version: 1.0 (Apple Message framework v752.2) 12, 22 -- In-Reply-To: {200611210017.kAL0HKv5004869-at-ns.microscopy.com} 12, 22 -- References: {200611210017.kAL0HKv5004869-at-ns.microscopy.com} 12, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 22 -- Message-Id: {0AAF81E5-6A33-4B38-8964-6739CAA635D9-at-arizona.edu} 12, 22 -- Content-Transfer-Encoding: 7bit 12, 22 -- From: David Elliott {Elliott-at-arizona.edu} 12, 22 -- Subject: Re: [Microscopy] AskAMicroscopist: staining multiple formvar/carbon coated slot 12, 22 -- Date: Tue, 21 Nov 2006 08:39:18 -0700 12, 22 -- To: jcraft-at-memphis.edu, Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 12, 22 -- X-Mailer: Apple Mail (2.752.2) 12, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
I still think in terms of Torr, so I had to convert your numbers over.
You say "high vacuum is allowed under a vacuum of 5x10-3 Pa". I convert that to about 4x10-5 Torr. You also say that you only get to 8x10-3 Pa when you open the manual valve between the column and the chamber. I convert that to 6x10-5 Torr.
That does sound like a poor vacuum. Our old JEOL 840A drops below 1x10-6 Torr on a good day with a diffusion pump. I think the safety interlock prevents operation above pressures of ~4x10-5 Torr. In that regard, the two microscopes seem consistent.
I agree with Chapman that this might indicate a leak in your chamber or perhaps you are not setting up the pumping properly. I have no idea what the proper procedure is for a Tescan. Our Hitachi S-2460N automates the process.
Regarding EDS, you should be able to use it quite well at 8x10-3 Pa. You will increasingly scatter the primary beam as the pressure increases. However, we do a lot of EDS work at 40 Pa (0.3 Torr). We simply have to take scattering into consideration. I greatly prefer "high" vacuum for quantitative EDS. It avoids complicating the spectrum of my analysis point with signal from the neighborhood.
Warren
-----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Tuesday, November 21, 2006 6:24 AM To: wesaia-at-iastate.edu
I agree that EBIC is not SC. In my case, it is a matter of conduction rather than true EBIC. I'm not looking at junctions, etc. The idea is to measure and image using specimen current and compliment with SE and or BSE. I image with SC and mix with SE or BSE and get quite interesting results.
In some cases, I'm using non-conductive specimens. In these cases, the beam current conductive portions have higher SC than areas that are non-conductive. However, in working with STEM specimens of IC cross sections, the metal conducts a certain amount while the passivation and ILD allows the beam to pass through. Depending on many factors, the SC can be radically different.
I suppose I'm just thinking at the keyboard. But I do wonder what the physics is behind all of this practical application.
gary g.
At 04:18 AM 11/21/2006, you wrote:
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==============================Original Headers============================== 13, 20 -- From gary-at-gaugler.com Tue Nov 21 16:58:13 2006 13, 20 -- Received: from qsmtp1.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 kALMwCaf002355 13, 20 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 16:58:13 -0600 13, 20 -- Received: (qmail 11399 invoked from network); 21 Nov 2006 14:58:06 -0800 13, 20 -- Received: by simscan 1.1.0 ppid: 11363, pid: 11396, t: 0.1894s 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 qsmtp1 with SMTP; 21 Nov 2006 14:58:06 -0800 13, 20 -- Message-Id: {7.0.1.0.2.20061121145655.025eaae0-at-gaugler.com} 13, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 13, 20 -- Date: Tue, 21 Nov 2006 14:58:05 -0800 13, 20 -- To: chad.parish-at-gmail.com 13, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 13, 20 -- Subject: Re: [Microscopy] Re: EBIC vs. SE 13, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 13, 20 -- In-Reply-To: {200611211218.kALCIn7D021934-at-ns.microscopy.com} 13, 20 -- References: {200611211218.kALCIn7D021934-at-ns.microscopy.com} 13, 20 -- Mime-Version: 1.0 13, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.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] Require Manual for HB601
Question: We have acquired a Vacuum Generators HB601 and would like to obtain instructions for alignment and other operating conditions. The manual that arrived with the instrument is good but someone out there might have written a more detailed alignment procedure. You can contact me offline if you wish.
This Question was submitted to Ask-A-Microscopist by (patrick.mccurdy-at-colostate.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, November 21, 2006 at 10:51:23 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both patrick.mccurdy-at-colostate.edu as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: patrick.mccurdy-at-colostate.edu Name: Pat McCurdy
Organization: Colorado State University
Education: Graduate College
Location: Fort Collins, CO, USA
Title: FE filiment lifetime
Question: According to Goldstein the life of a field emission Schotky filament is ~1 year. JEOL's service manual also states the lifetime of the emitter is ~1 year. The reason stated by both sources is the limited reservoir of zirconia surrounding the tip, which lowers the work function. Our emitter is now almost 5 years old. Why the discrepancy?
That is an interesting question. What I see is about two years of life. However, the variables are brand of FE tip and gun chamber vacuum and duty cycle of gun on/off. My gun chamber vacuum is about 1.5E-10 Torr.
For a Denka tip on all the time, I am seeing at least two years. The following SEM pix shows a Denka tip at two years and there is plenty of ZrO2. There have been FEI tips in my SEM but none worked. The way Zeiss does this is to cull out the losers (Denka and FEI) and only deliver the ones that pass their scrutiny. It does make a difference. I'm not sure what criteria they use but it works.
http://www.gaugler.com/fe%20tip-2.jpg
I was prepared for a tip replacement every year but do not experience this need at all. The gun is on 24/7. It is very stable and unflinching in Iext. This is in a Zeiss Supra 55VP.
Disclaimer: No financial interest in Zeiss, Denka or FEI other than to keep Zeiss supporting my Supra 55VP.
gary g.
At 05:40 PM 11/21/2006, you wrote:
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==============================Original Headers============================== 14, 20 -- From gary-at-gaugler.com Tue Nov 21 20:59:28 2006 14, 20 -- Received: from qsmtp3.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 kAM2xSjS006453 14, 20 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 20:59:28 -0600 14, 20 -- Received: (qmail 31868 invoked from network); 21 Nov 2006 18:59:27 -0800 14, 20 -- Received: by simscan 1.1.0 ppid: 31817, pid: 31866, t: 0.1798s 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 qsmtp3 with SMTP; 21 Nov 2006 18:59:27 -0800 14, 20 -- Message-Id: {7.0.1.0.2.20061121184818.025be488-at-gaugler.com} 14, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 14, 20 -- Date: Tue, 21 Nov 2006 18:59:21 -0800 14, 20 -- To: patrick.mccurdy-at-colostate.edu 14, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 14, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: FE filiment lifetime 14, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 14, 20 -- In-Reply-To: {200611220140.kAM1eiwf021916-at-ns.microscopy.com} 14, 20 -- References: {200611220140.kAM1eiwf021916-at-ns.microscopy.com} 14, 20 -- Mime-Version: 1.0 14, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
We have just installed our first Schottky FE (Hitachi 4300 SE/N.) There is plenty of current, and therefore a bit of temptation to wind the filament temperature back just a little to - maybe? - increase lifetime and resolution. Has any one tried this, does anyone know how the optimal temperature is determined? ZrO2 mobility, gas absorption, electron emission, energy spread? ...are there other factors?
Sally
Dr SJ Stowe Facility Coordinator ANU Electron Microscopy Unit ANU CRICOS#00120C
} -----Original Message----- } From: gary-at-gaugler.com [mailto:gary-at-gaugler.com] } Sent: Wednesday, 22 November 2006 2:00 PM } To: sally.stowe-at-anu.edu.au } Subject: [Microscopy] Re: AskAMicroscopist: FE filiment lifetime } } } } } } --------------------------------------------------------------- } ------------- } The Microscopy ListServer -- CoSponsor: The Microscopy } Society of America To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver
==============================Original Headers============================== 10, 26 -- From sally.stowe-at-anu.edu.au Tue Nov 21 23:28:27 2006 10, 26 -- Received: from mail.rsbs.anu.edu.au (mail.rsbs.anu.edu.au [150.203.72.70]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAM5SQXO020020 10, 26 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 23:28:26 -0600 10, 26 -- Received: from rsbs2262 (rsbs2262.rsbs.anu.edu.au [150.203.36.144]) 10, 26 -- by mail.rsbs.anu.edu.au (Postfix) with ESMTP 10, 26 -- id C6360F44E6A; Wed, 22 Nov 2006 16:28:14 +1100 (EST) 10, 26 -- Reply-To: {sally.stowe-at-anu.edu.au} 10, 26 -- From: "Sally Stowe" {sally.stowe-at-anu.edu.au} 10, 26 -- To: {gary-at-gaugler.com} , {microscopy-at-microscopy.com} 10, 26 -- Subject: RE: [Microscopy] Re: AskAMicroscopist: FE filiment lifetime 10, 26 -- Date: Wed, 22 Nov 2006 16:28:14 +1100 10, 26 -- Message-ID: {004b01c70df7$022e4b20$9024cb96-at-rsbs.anu.edu.au} 10, 26 -- MIME-Version: 1.0 10, 26 -- Content-Type: text/plain; 10, 26 -- charset="us-ascii" 10, 26 -- Content-Transfer-Encoding: 7bit 10, 26 -- X-Priority: 3 (Normal) 10, 26 -- X-MSMail-Priority: Normal 10, 26 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 10, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1506 10, 26 -- Importance: Normal 10, 26 -- In-Reply-To: {200611220259.kAM2xdeX006634-at-ns.microscopy.com} 10, 26 -- X-RSBS-MailScanner-Information: Please contact the ISP for more information 10, 26 -- X-RSBS-MailScanner: Found to be clean 10, 26 -- X-MailScanner-From: sally.stowe-at-anu.edu.au ==============================End of - Headers==============================
I wanted to thank you all for the numerous answers I got for my problem. Well I still had to go through the whole alignment procedure(what I wanted to avoid by mailing this list - I thought it was possible through the direct alignments) but the problem is fixed. Many thanks to the FEI people for their care.
Stephane
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==============================Original Headers============================== 6, 20 -- From nizets2-at-yahoo.com Wed Nov 22 06:41:50 2006 6, 20 -- Received: from web37401.mail.mud.yahoo.com (web37401.mail.mud.yahoo.com [209.191.91.133]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAMCfnwk007431 6, 20 -- for {microscopy-at-microscopy.com} ; Wed, 22 Nov 2006 06:41:49 -0600 6, 20 -- Received: (qmail 70474 invoked by uid 60001); 22 Nov 2006 12:41:47 -0000 6, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 20 -- s=s1024; d=yahoo.com; 6, 20 -- h=X-YMail-OSG:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 6, 20 -- b=C4oCxW3ghfm43r471SAVrnUKUd2vWPL0fpMc8aiYp7dST7jYaUHv1x31rSNBaezgpEMFNElJvKcUOHVhS73VfK9EoClf7F4uL4k1aSrnBnxFEPR6spIcj8v00mf6bq9yppzdKHy/ghwrrDEEBPoc/HbgYZs86v9eKcFBU1xghnM=; 6, 20 -- X-YMail-OSG: RbeIZuMVM1kf5RSAkb2d2u3j_lPRaQXsDTuIf2AD 6, 20 -- Received: from [80.122.101.102] by web37401.mail.mud.yahoo.com via HTTP; Wed, 22 Nov 2006 04:41:47 PST 6, 20 -- Date: Wed, 22 Nov 2006 04:41:47 -0800 (PST) 6, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 20 -- Subject: RE: [Microscopy] alignment with tecnai twin G20 - Thanks 6, 20 -- To: microscopy-at-microscopy.com 6, 20 -- In-Reply-To: {00948E5E66F1374FB9284A7B78138EB806E6CAD7-at-hlexc03.w2k.feico.com} 6, 20 -- MIME-Version: 1.0 6, 20 -- Content-Type: text/plain; charset=iso-8859-1 6, 20 -- Content-Transfer-Encoding: 8bit 6, 20 -- Message-ID: {481904.70280.qm-at-web37401.mail.mud.yahoo.com} ==============================End of - Headers==============================
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Title-Subject: [Filtered] Insulin and glucagon staining in the pancreas
Question: Hi All. I was wanting to do some immunohistochemistry on paraffin pancreas sections. I would like to double stain the sections with insulin and glucagon, then count the number of islets relative to the whole area of the pancreas. Has anyone does such staining before using DAB? Thanks in advance, Emily
Sam, My experience is that Larry is absolutely correct. You'll be starting at room temperature and the temperature is likely to rise considerably and rapidly at the beam. On the other hand, holding a specimen below 4 C wouldn't be very difficult using a thermoelectric (Peltier) device. All that is needed is an electrical feedthrough (2 connections for the device and an optional 2 connections for a thermocouple to read temperature and/or provide feedback to an automatic controller) and a heatsink connection from the hot side of the device to the wall of the stage or chamber (several pieces of ground braid can provide a flexible thermal connection). This is assuming that you want to be able to vary the temperature at will.
On the other hand, if all you want to do is immobilize your sample either for a quick look, or to watch what happens as the sample becomes mobilized, than I would think that chilling the Quantomix capsule by some amount would take care of that. How much would most likely have to be determined by experiment.
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: larry-at-celtic.freewire.co.uk [mailto:larry-at-celtic.freewire.co.uk] Sent: Tuesday, November 14, 2006 4:08 PM To: kenconverse-at-qualityimages.biz
} ----------------------------------------------------------------------- } ----- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America I guess you actually mean the temperature under the beam ..... The temperature in the specimen chamber generally is going to be close to ambient (except for an ESEM, were the sample will be ~5 deg C)
Key things to consider are probe current, scan speed, magnification and thermal conduction path away from the imaged area. In the case of Quantomix capsules, with a FEG-SEM it is relatively easy to punch holes in the membrane with the beam, rather less so with W SEMs.
It is also relatively easy in most SEMs to find conditions which will melt lower temperature thermosetting polymers, so local sample temperatures in the 50 to 100 deg C aren't too difficult to generate, with the right sample.
Basically, I think there are so many variables involved, you're just going to have to try it.
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==============================Original Headers============================== 6, 16 -- From larry-at-celtic.freewire.co.uk Tue Nov 14 15:04:26 2006 6, 16 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 6, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAEL4Nue017767 6, 16 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 14 Nov 2006 15:04:26 -0600 6, 16 -- Received: from [194.164.78.253] (modem-rack4-modem253.netkonect.net [194.164.78.253]) 6, 16 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id kAEL2b3B004597; 6, 16 -- Tue, 14 Nov 2006 21:03:31 GMT 6, 16 -- Mime-Version: 1.0 6, 16 -- Message-Id: {p06210202c17fd7b6ceb4-at-[194.164.78.26]} 6, 16 -- In-Reply-To: {200611141429.kAEETu5l019725-at-ns.microscopy.com} 6, 16 -- References: {200611141429.kAEETu5l019725-at-ns.microscopy.com} 6, 16 -- Date: Tue, 14 Nov 2006 20:38:18 +0000 6, 16 -- To: Sam.Murray-at-port.ac.uk, Microscopy-at-MSA.Microscopy.Com 6, 16 -- X-from: Larry Stoter {larry-at-celtic.freewire.co.uk} 6, 16 -- Subject: Re: [Microscopy] Temperature inside SEM chamber? 6, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
==============================Original Headers============================== 22, 28 -- From kenconverse-at-qualityimages.biz Wed Nov 22 09:16:09 2006 22, 28 -- Received: from dpmailmta02.doteasy.com (dpmailmta02.doteasy.com [65.61.209.84]) 22, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAMFG8BX032011 22, 28 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 22 Nov 2006 09:16:09 -0600 22, 28 -- Received: from qualityimages.biz (unverified [192.168.101.16]) 22, 28 -- by dpmailmta02.doteasy.com (DEO) with ESMTP id 22945706-1814644 22, 28 -- for multiple; Wed, 22 Nov 2006 07:29:24 -0800 22, 28 -- Received: from Ken [76.179.237.214] by qualityimages.biz with ESMTP 22, 28 -- (SMTPD32-8.05) id A99A2470218; Wed, 22 Nov 2006 07:15:38 -0800 22, 28 -- From: "Ken Converse" {kenconverse-at-qualityimages.biz} 22, 28 -- To: {larry-at-celtic.freewire.co.uk} , 22, 28 -- "MSA Listserver" {Microscopy-at-MSA.Microscopy.Com} 22, 28 -- Subject: RE: [Microscopy] Re: Temperature inside SEM chamber? 22, 28 -- Date: Wed, 22 Nov 2006 10:15:02 -0500 22, 28 -- Message-ID: {007701c70e49$08583b30$6401a8c0-at-Ken} 22, 28 -- MIME-Version: 1.0 22, 28 -- Content-Type: text/plain; 22, 28 -- charset="us-ascii" 22, 28 -- X-Priority: 3 (Normal) 22, 28 -- X-MSMail-Priority: Normal 22, 28 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 22, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 22, 28 -- Importance: Normal 22, 28 -- In-Reply-To: {200611142108.kAEL85ZC022598-at-ns.microscopy.com} 22, 28 -- X-IP-stats: Incoming Last 0, First 55, in=1250933, out=0, spam=0 ip=192.168.101.16 22, 28 -- X-External-IP: 192.168.101.16 22, 28 -- Content-Transfer-Encoding: 8bit 22, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAMFG8BX032011 ==============================End of - Headers==============================
Our JEOL FETEM is on the third emitter in over nine years. We replaced one when moving the microscope (-at- three years), and another failed three years later. We believe JEOL has just been very conservative in rating the tip life. Five years, though, is extraordinary.
John Mardinly Intel
This is the opinion of the author and does not represent the position of Intel Corporation.
-----Original Message----- X-from: patrick.mccurdy-at-colostate.edu [mailto:patrick.mccurdy-at-colostate.edu] Sent: Tuesday, November 21, 2006 5:39 PM To: Mardinly, John
This Question was submitted to Ask-A-Microscopist by (patrick.mccurdy-at-colostate.edu) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, November 21, 2006 at 10:51:23 Remember to consider the Grade/Age of the student when considering the Question ------------------------------------------------------------------------ --- Please reply to both patrick.mccurdy-at-colostate.edu as well as to the Microscopy Listserver ------------------------------------------------------------------------ ---
Email: patrick.mccurdy-at-colostate.edu Name: Pat McCurdy
Organization: Colorado State University
Education: Graduate College
Location: Fort Collins, CO, USA
Title: FE filiment lifetime
Question: According to Goldstein the life of a field emission Schotky filament is ~1 year. JEOL's service manual also states the lifetime of the emitter is ~1 year. The reason stated by both sources is the limited reservoir of zirconia surrounding the tip, which lowers the work function. Our emitter is now almost 5 years old. Why the discrepancy?
The emitter on our JEOL FETEM is now nearly 9 years old having been installed in January 1998 (71,763hrs). We operate at a lower emission than JEOL usually use (~110microA) in order to get reasonable EELS resolution and enough beam current for Z contrast STEM.
Regards
Alan
At 10:26 AM 11/22/2006 -0600, john.mardinly-at-intel.com wrote:
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Alan W Nicholls, PhD Director of Research Service Facility (Electron Microscopy) Research Resources Center - East (M/C 337) Room 100 Science and Engineering South Building The University of Illinois at Chicago 845 West Taylor St Chicago, IL 60607-7058
} -----Original Message----- } From: gary-at-gaugler.com [mailto:gary-at-gaugler.com] } Sent: Tuesday, November 21, 2006 9:00 PM } To: Dusevich, Vladimir } Subject: [Microscopy] Re: AskAMicroscopist: FE filiment lifetime } } } } } -------------------------------------------------------------- } -------------- } 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 } -------------------------------------------------------------- } -------------- } } That is an interesting question. What I see is about two years of } life. However, the variables are brand of FE tip and gun chamber } vacuum and duty cycle of gun on/off. My gun chamber vacuum is about } 1.5E-10 Torr. } } For a Denka tip on all the time, I am seeing at least two years. The } following SEM pix shows a Denka tip at two years and there is plenty } of ZrO2. There have been FEI tips in my SEM but none worked. The way
} Zeiss does this is to cull out the losers (Denka and FEI) and only } deliver the ones that pass their scrutiny. It does make a difference. } I'm not sure what criteria they use but it works. } } http://www.gaugler.com/fe%20tip-2.jpg } } I was prepared for a tip replacement every year but do not experience } this need at all. The gun is on 24/7. It is very stable and } unflinching in Iext. This is in a Zeiss Supra 55VP. } } Disclaimer: No financial interest in Zeiss, Denka or FEI other than } to keep Zeiss supporting my Supra 55VP. } } gary g. } } }
==============================Original Headers============================== 8, 23 -- From DusevichV-at-umkc.edu Wed Nov 22 13:46:06 2006 8, 23 -- Received: from kc-msxproto1.kc.umkc.edu (kc-msxproto1.kc.umkc.edu [134.193.143.167]) 8, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAMJk6JG007769 8, 23 -- for {microscopy-at-msa.microscopy.com} ; Wed, 22 Nov 2006 13:46:06 -0600 8, 23 -- Received: from KC-MSX1.kc.umkc.edu ([134.193.32.11]) by kc-msxproto1.kc.umkc.edu with Microsoft SMTPSVC(6.0.3790.1830); 8, 23 -- Wed, 22 Nov 2006 13:46:06 -0600 8, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 8, 23 -- Content-class: urn:content-classes:message 8, 23 -- MIME-Version: 1.0 8, 23 -- Content-Type: text/plain; 8, 23 -- charset="us-ascii" 8, 23 -- Subject: FW: [Microscopy] Re: AskAMicroscopist: FE filiment lifetime 8, 23 -- Date: Wed, 22 Nov 2006 13:46:06 -0600 8, 23 -- Message-ID: {032EC4F75A527A4FA58C5B1B5DECFBB33ADC5E-at-KC-MSX1.kc.umkc.edu} 8, 23 -- X-MS-Has-Attach: 8, 23 -- X-MS-TNEF-Correlator: 8, 23 -- Thread-Topic: [Microscopy] Re: AskAMicroscopist: FE filiment lifetime 8, 23 -- Thread-Index: AccN4lUdrO923mWuQCCZw9FJyaaOhgAZQarwAAnVnTA= 8, 23 -- From: "Dusevich, Vladimir" {DusevichV-at-umkc.edu} 8, 23 -- To: {microscopy-at-msa.microscopy.com} 8, 23 -- X-OriginalArrivalTime: 22 Nov 2006 19:46:06.0102 (UTC) FILETIME=[D9776360:01C70E6E] 8, 23 -- Content-Transfer-Encoding: 8bit 8, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAMJk6JG007769 ==============================End of - Headers==============================
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Email: bairdgb-at-uwec.edu Name: Graham Baird
Organization: University of Wisconsin-Eau Claire
Title-Subject: [Filtered] Backscatter detector for Hatachi S510 SEM
Question: Does anyone know how much a backscatter detector for a Hatachi S510 SEM could cost? Or could even send me a quote? Thanks a lot. Cheers, Graham Baird UWEC Geology Dept.
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Email: wa5ekh-at-juno.com Name: Charles Jeffrey Day
Organization: none
Title-Subject: [Filtered] Electroscan E3 Environmental SEM and Tracor Be-Window Xray Detector
Question: Last reminder: The company I work for has descided to buy a new SEM, so they will soon sell all or part of this ESEM and Xray system (1 board in each needs repair).You can contact me by direct email.(I have no financial interest in this instrument, but I know how hard it is to find parts for these, so this may have some of the parts you need..)
EBS is the US rep for KE Developments. SPI is the US rep for Robinson. If I recall correctly, a Robinson Model 8 is about $10K, depending on options.
gary g.
At 03:50 PM 11/22/2006, you wrote:
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==============================Original Headers============================== 8, 21 -- From gary-at-gaugler.com Wed Nov 22 18:14:53 2006 8, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kAN0Erps024924 8, 21 -- for {microscopy-at-microscopy.com} ; Wed, 22 Nov 2006 18:14:53 -0600 8, 21 -- Received: (qmail 29419 invoked from network); 22 Nov 2006 16:14:52 -0800 8, 21 -- Received: by simscan 1.1.0 ppid: 29398, pid: 29417, t: 0.1700s 8, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 8, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 8, 21 -- by qsmtp3 with SMTP; 22 Nov 2006 16:14:52 -0800 8, 21 -- Message-Id: {7.0.1.0.2.20061122161252.025d4390-at-gaugler.com} 8, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 8, 21 -- Date: Wed, 22 Nov 2006 16:14:47 -0800 8, 21 -- To: bairdgb-at-uwec.edu 8, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 8, 21 -- Subject: Re: [Microscopy] viaWWW: Backscatter detector for Hatachi S510 8, 21 -- SEM 8, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 8, 21 -- In-Reply-To: {200611222350.kAMNojfg008234-at-ns.microscopy.com} 8, 21 -- References: {200611222350.kAMNojfg008234-at-ns.microscopy.com} 8, 21 -- Mime-Version: 1.0 8, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Roberto Garcia, Fred Stevie, Phil Russell and there may be more authors wrote a how-to-do this paper. They are in the Materials Science Dept at North Carolina State University. Unfortunately, I can't lay my hands on the paper or I would give you the proper citation. Basically, they use a puck that is compatible with the SIMS instrument and controllably grind the edges of a sample at the same angles on four corners. They measure the thickness of layers revealed at the corners to make their tilt corrections to find the polishing surface parallel to the backside and then grind away. When they get close, they use interference fringes to make their final tilt corrections. They use a competitor's product, but I believe that the same action could easily be accomplished in the same fashion with a Tripod PolisherR and at a significantly lower cost.
Disclaimer: South Bay Technology, Inc. makes and sells the Tripod PolisherR
-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: jyw-at-lge.com [mailto:jyw-at-lge.com] Sent: Monday, November 20, 2006 6:58 PM To: Walck-at-SouthBayTech.com
Hello
I need to prepare the LED chip for back side SIMS anaysis. SIMS depth profile resolution is significantly degraded if a large depth must be sputtered prior to the depth of analyticall interest. This problem can be exacerbated on LED chips which is covered thick electrode. When the LED chip sputtering which has thick eletrode exhibit strong nonuniform sputtering properties . These problems can be avoided if sufficient sample substrate material can be removed with adequate precision to allow SIMS depth profiling from the sample back side.
Does anyone have experience or suggestions on how to prepare back side SIMS samples? Method, Recipe for polishing step and so on..
Thanks in advance
Young Woo Jeong
==============================Original Headers============================== 10, 22 -- From jyw-at-lge.com Mon Nov 20 20:54:28 2006 10, 22 -- Received: from mail1.lge.co.kr (mail1.lge.co.kr [156.147.1.151]) 10, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAL2sR55029263 10, 22 -- for {Microscopy-at-microscopy.com} ; Mon, 20 Nov 2006 20:54:28 -0600 10, 22 -- Received: from [156.147.51.54] (jyw-at-lge.com) by 10, 22 -- mail1.lge.co.kr (Terrace MailWatcher) 10, 22 -- with ESMTP id 2006112111:54:22:863195.12230.155 10, 22 -- for {Microscopy-at-microscopy.com} ; 10, 22 -- Tue, 21 Nov 2006 11:54:22 +0900 (KST) 10, 22 -- Subject: TEM sample preparation for back side SIMS analysis 10, 22 -- Reply-To: "=?EUC-KR?B?waS/tb/s?=" {jyw-at-lge.com} 10, 22 -- From: "=?EUC-KR?B?waS/tb/s?=" {jyw-at-lge.com} 10, 22 -- To: Microscopy-at-microscopy.com 10, 22 -- Message-ID: {OF8DE4D82F.1F7B65DD-ON4925722D.000FF6CD-4925722D.000FF6CF-at-lge.com} 10, 22 -- Date: Tue, 21 Nov 2006 11:54:22 +0900 10, 22 -- Return-Receipt-To: "=?EUC-KR?B?waS/tb/s?=" {jyw-at-lge.com} 10, 22 -- X-MIMETrack: Serialize by Router on LGEKRHQMS03/LGE/LG Group(Release 6.5.4FP2HF559 | September 10, 22 -- 7, 2006) at 2006/11/21 11:54:22 10, 22 -- MIME-Version: 1.0 10, 22 -- Content-type: text/plain; charset=US-ASCII 10, 22 -- X-TERRACE-SPAMMARK: NOT spam-marked. 10, 22 -- (by Terrace) ==============================End of - Headers==============================
==============================Original Headers============================== 19, 22 -- From walck-at-southbaytech.com Wed Nov 22 20:23:20 2006 19, 22 -- Received: from ylpvm12.prodigy.net (ylpvm12-ext.prodigy.net [207.115.57.43]) 19, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAN2NK5L011211 19, 22 -- for {Microscopy-at-microscopy.com} ; Wed, 22 Nov 2006 20:23:20 -0600 19, 22 -- X-ORBL: [64.169.193.90] 19, 22 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 19, 22 -- by ylpvm12.prodigy.net (8.13.7 out spool5000 dk/8.13.7) with ESMTP id kAN2MYYM027762; 19, 22 -- Wed, 22 Nov 2006 21:22:35 -0500 19, 22 -- From: "Scott Walck" {walck-at-southbaytech.com} 19, 22 -- To: {Microscopy-at-microscopy.com} 19, 22 -- Cc: {jyw-at-lge.com} 19, 22 -- Subject: RE: [Microscopy] TEM sample preparation for back side SIMS analysis 19, 22 -- Date: Wed, 22 Nov 2006 18:23:41 -0800 19, 22 -- Message-ID: {00aa01c70ea6$64e75270$7801a8c0-at-dynamicbl8uno3} 19, 22 -- MIME-Version: 1.0 19, 22 -- Content-Type: text/plain; 19, 22 -- charset="us-ascii" 19, 22 -- Content-Transfer-Encoding: 7bit 19, 22 -- X-Mailer: Microsoft Office Outlook 11 19, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 19, 22 -- Thread-Index: AccNGM/MlHmt7yWYRGSN6cTiH8iVoAAelYBA 19, 22 -- In-Reply-To: {200611210257.kAL2vfXA002764-at-ns.microscopy.com} ==============================End of - Headers==============================
Graham Baird wrote: =================================================== Does anyone know how much a backscatter detector for a Hatachi S510 SEM could cost? Or could even send me a quote? Thanks a lot. ================================================== Full information about the Robinson backscattered electron detector can be found on URL http://2spi.com/catalog/instruments/robinson-backscattered-electron-BSE-detector.shtml
There is an on-line questionnaire that can be filled out and once we have the details of your installation and requirements, we can give you a recommendation and firm price quote.
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.
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==============================Original Headers============================== 10, 26 -- From cgarber-at-2spi.com Wed Nov 22 22:45:10 2006 10, 26 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 10, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAN4jArA027629 10, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 22 Nov 2006 22:45:10 -0600 10, 26 -- Received: from ibm1x23g2abfyg (000-191-484.area3.spcsdns.net [68.246.233.20] (may be forged)) 10, 26 -- (authenticated bits=0) 10, 26 -- by diskless5.axs2000.net (8.12.11/8.12.11) with ESMTP id kAN4j2s9021132 10, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 22 Nov 2006 23:45:09 -0500 10, 26 -- X-IDV-FirstRcvd: 000-191-484.area3.spcsdns.net [68.246.233.20] (may be forged) 10, 26 -- X-IDV-HELO: ibm1x23g2abfyg 10, 26 -- X-IDV-Authenticated-User: cgarber 10, 26 -- Message-ID: {00b501c70eba$244ab450$14e9f644-at-ibm1x23g2abfyg} 10, 26 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 10, 26 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 10, 26 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 10, 26 -- Subject: Backscattered electron detectors 10, 26 -- Date: Wed, 22 Nov 2006 23:44:20 -0500 10, 26 -- MIME-Version: 1.0 10, 26 -- Content-Type: text/plain; 10, 26 -- charset="Windows-1252" 10, 26 -- X-Priority: 3 10, 26 -- X-MSMail-Priority: Normal 10, 26 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 10, 26 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 10, 26 -- Content-Transfer-Encoding: 8bit 10, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAN4jArA027629 ==============================End of - Headers==============================
I finally found the necessary sequence to have "green light" for "high vacuum" mode. Funnily, the vacuum remains the same, but now I have a green signal and I can activate the HT and filament! Here are some few more questions:
1. In TEM one needs to tilt the sample to improve the signal by EDX. Is it the same in SEM? (considering that the detector is also at an angle above the sample) 2. Is it realistic to use HT of 5kV or 10kV to take EDX spectra by SEM? 3. Resolution issue: while observing mineral particules (quarz for example), when I reach a magnification of 2kx or higher (to observe particles several µm big for example), the image is not sharp. Using the mode which offers the maximum resolution (cleverly called “resolution mode”), the beam reaches a size of 60nm. In these conditions it is not surprising that the image is not sharp. The SEM allows magnification of up to 500kx!! What is the point to use so high magnification if the image is blurred? What is the expected resolution of a SEM?
Stephane
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==============================Original Headers============================== 7, 19 -- From nizets2-at-yahoo.com Thu Nov 23 09:39:03 2006 7, 19 -- Received: from web37415.mail.mud.yahoo.com (web37415.mail.mud.yahoo.com [209.191.91.147]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kANFd2jH020698 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 23 Nov 2006 09:39:03 -0600 7, 19 -- Received: (qmail 87141 invoked by uid 60001); 23 Nov 2006 15:39:02 -0000 7, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 19 -- s=s1024; d=yahoo.com; 7, 19 -- h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 7, 19 -- b=hVWsNBLdPT8OJ+cRYh7ebPum4IIFJrBmR3iGwec0wlKHA9DoHA/QWcz0Ny0TO9FKzCQRrE12i3MJSOdXHJM4OVfZ4rYzpf0z9rcmRh/NyJ3UWHQk9oIbGFPSeiECDSFAEmz9nmscWmfDHnv7NmVV0umhPc2YuFBrC2FUoLNlOJE=; 7, 19 -- X-YMail-OSG: XhFNP5wVM1mltPNRJpdw3q2cRh2TsqpyvuUNWwrM4nNJqo0e9k0iew.rE6kvTC5T8Ni3NYra.tcG2iUuqjZe7gs1DNcPR43b1HKoYNNKJeY93sCBEktlIJhBjuNmsOCDwjsJ2PAXT_qUFfM- 7, 19 -- Received: from [80.122.101.102] by web37415.mail.mud.yahoo.com via HTTP; Thu, 23 Nov 2006 07:39:02 PST 7, 19 -- Date: Thu, 23 Nov 2006 07:39:02 -0800 (PST) 7, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 19 -- Subject: Re: Basic SEM functions 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: text/plain; charset=iso-8859-1 7, 19 -- Content-Transfer-Encoding: 8bit 7, 19 -- Message-ID: {292322.86779.qm-at-web37415.mail.mud.yahoo.com} ==============================End of - Headers==============================
First off, is there an ion pump at the top of the column for the gun chamber? If there is, the I would assume that the vacuum sequence would be to open the isolation valve and let everything reach terminal vacuum, then close the isolation valve and turn on the gun chamber ion pump to achieve highest vacuum for the gun. If the SEM is of traditional design, it will have one or more differential pumping apertures in the column such that the gun will always be at high vac regardless of what is going on in the chamber. Low vac for the gun will likely kill it quickly.
1. If the EDS detector is tilted, the specimen can be horizontal. The effective take off angle (mechanical tilt + specimen tilt) will affect total counts per second at a particular filter time.
2. Yes. In some cases, it is required. This is a very specimen-specific issue. For example, if you are looking at high Z specimens, then KV needs to be about twice the highest Ma or La eV value. Take Cu for example. It has Ka peak at 8.040KeV and La peak at .930eV. However, Pr has an Ma peak at .929eV. It's unlikely that you would have Cu and Pr but point is that with double peak elements like this example, you would need about 18KV beam to bring up both Cu peaks to ensure Cu is what you have and for the system to quant. A smart system will complain that KV is too low for quant.
On the other hand, if you are looking at light elements like C, O, N, F, Al, these only have one peak of interest--Ka. F is especially tricky since in normal form, it will be burned by a strong beam at high KV. Thus, 5KV is a good value to use to avoid specimen damage, reduce volumetric interaction and still get enough counts to do quant. In IC processing, F is a frequently used gas. It used with Si and Al chucks and is supposed to be in some areas and not in others. If the KV is too high, the very thin layer of F will be overwhelmed by a high KV beam such that the layer is pierced and therefore producing a large peak at Si or Al and likely missing the F.
3. How do you know the beam is 60nm? Apart from this, does the system stigmate properly? Is the gun centered? Is the final aperture centered? Is the condenser lens open or reduced?
As far as SEM resolution is concerned, you have to ask what the expected resolution of "your" SEM should be. Resolution changes from maker-to-maker and model-to-model. It also changes with WD. Shorter WD, better resolution but at the expense of less SEs. FESEMs will have better resolution than thermionic SEMs. There are a whole bunch of variables that affect resolution. Another issue is size of final aperture. Smaller, better but reduced S/N.
Hope this helps.
gary g.
At 07:42 AM 11/23/2006, you wrote:
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==============================Original Headers============================== 15, 22 -- From gary-at-gaugler.com Thu Nov 23 11:17:57 2006 15, 22 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 15, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kANHHupW000751 15, 22 -- for {microscopy-at-microscopy.com} ; Thu, 23 Nov 2006 11:17:56 -0600 15, 22 -- Received: (qmail 12763 invoked from network); 23 Nov 2006 09:17:56 -0800 15, 22 -- Received: by simscan 1.1.0 ppid: 12735, pid: 12761, t: 0.2101s 15, 22 -- scanners: regex: 1.1.0 attach: 1.1.0 15, 22 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 15, 22 -- by qsmtp2 with SMTP; 23 Nov 2006 09:17:56 -0800 15, 22 -- Message-Id: {7.0.1.0.2.20061123084900.024eac80-at-gaugler.com} 15, 22 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 15, 22 -- Date: Thu, 23 Nov 2006 09:17:51 -0800 15, 22 -- To: nizets2-at-yahoo.com 15, 22 -- From: Gary Gaugler {gary-at-gaugler.com} 15, 22 -- Subject: Re: [Microscopy] Re: Basic SEM functions 15, 22 -- Cc: MSA listserver {microscopy-at-microscopy.com} 15, 22 -- In-Reply-To: {200611231542.kANFgDEf023695-at-ns.microscopy.com} 15, 22 -- References: {200611231542.kANFgDEf023695-at-ns.microscopy.com} 15, 22 -- Mime-Version: 1.0 15, 22 -- Content-Type: text/plain; charset="iso-8859-1"; format=flowed 15, 22 -- Content-Transfer-Encoding: 8bit 15, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kANHHupW000751 ==============================End of - Headers==============================
I have marked my comments against your email below
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: {nizets2-at-yahoo.com} To: {protrain-at-emcourses.com} Sent: Thursday, November 23, 2006 3:40 PM
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Organization: Max-Planck-Institute of Biochemistry
Title-Subject: [Filtered] How to generate a simple excitation pattern with a fluorescent microscope
Question: Hi, all,
I am working with photoactivatable (ìcagedî) compounds and I am trying to create a simple pattern (a smiley face or for example the letter ëAí) of uncaged compound within a brain slice. This is just supposed to be a prove-of-principle to show the power and spatial resolution of photoactivation. So far, Iíve been using a 10x objective and a DAPI filter set on an old Zeiss Axiophot for uncaging. My idea was to stick a small blackened disc (with air-slits showing the respective pattern) in the field stop of the microscope so that the pattern should in principle be projected onto the brain slice as DAPI light. I had our machine shop make me 3 different discs with slit widths (length:1cm) of 200µm, 600µm, or 1mm to test this idea - but little success. If I use any of these discs to look at a more or less homogenous fluorescent sample, I only see a dimmed image of the entire sample but never a crisp bar of excitation/fluorescence across the specimen. I am wondering now if there is a conceptual mistake in my approach or if I am just not using the proper slits - basically I have no idea whatís going on. So Iíd be thrilled if someone could enlighten me! Also, is there a company out there that could produce discs with a little more elaboration such as a smiley face ? Thanks in advance.
I realize the question basically concerns EDX, but there is also one thing when observing the morphology of a tilted specimen. I ALWAYS advise against the of use the tilt-correction control, or you will get distortion: for example, it makes spheres (which should appear circular when tilted at any angle) come out as ellipsoids, and rectangular blocks will appear as parallelepipeds (lovely word!).
----------------------------------- Robert H. Olley URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
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==============================Original Headers============================== 6, 23 -- From hinmeigeng-at-hotmail.com Fri Nov 24 09:48:37 2006 6, 23 -- Received: from bay0-omc2-s37.bay0.hotmail.com (bay0-omc2-s37.bay0.hotmail.com [65.54.246.173]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAOFmaFY026548 6, 23 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 24 Nov 2006 09:48:36 -0600 6, 23 -- Received: from hotmail.com ([65.55.130.89]) by bay0-omc2-s37.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 23 -- Fri, 24 Nov 2006 07:48:36 -0800 6, 23 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 6, 23 -- Fri, 24 Nov 2006 07:48:35 -0800 6, 23 -- Message-ID: {BAY125-F93FA77AEBAFE45FDB7E35CAE10-at-phx.gbl} 6, 23 -- Received: from 65.55.130.123 by by125fd.bay125.hotmail.msn.com with HTTP; 6, 23 -- Fri, 24 Nov 2006 15:48:30 GMT 6, 23 -- X-Originating-IP: [134.225.1.161] 6, 23 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 6, 23 -- X-Sender: hinmeigeng-at-hotmail.com 6, 23 -- Reply-To: R.H.Olley-at-reading.ac.uk 6, 23 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 6, 23 -- To: Microscopy-at-MSA.Microscopy.Com 6, 23 -- Cc: hinmeigeng-at-hotmail.com, nizets2-at-yahoo.com 6, 23 -- Subject: Basic SEM functions -Tilt 6, 23 -- Date: Fri, 24 Nov 2006 15:48:30 +0000 6, 23 -- Mime-Version: 1.0 6, 23 -- Content-Type: text/plain; format=flowed 6, 23 -- X-OriginalArrivalTime: 24 Nov 2006 15:48:35.0728 (UTC) FILETIME=[00686100:01C70FE0] ==============================End of - Headers==============================
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Email: maloneyb-at-fiu.edu Name: Barbara Maloney
Organization: FIU
Title-Subject: [Filtered] Phillips 300 TEM
Question: Dear Group - I know everyone here in the US is probably on Thanksgiving holiday, but by chance anyone have an idea what year the Phillip 300 TEM was produced? It would be great if one could respond to this query ASAP - also anyone have any idea how many 300s are still operational? Thanks so much. Barbara
The Philips EM 300 was introduced in 1966. Versions of the EM300 with STEM were first sold in 1968.
Information found in "The "History of Electron MIcroscopes - 1986" from 11th International Congress on EM-Kyoto 1986
Nestor Your Friendly Neighborhood SysOp
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==============================Original Headers============================== 8, 13 -- From zaluzec-at-microscopy.com Sat Nov 25 08:14:45 2006 8, 13 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 8, 13 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAPEEjMC005395; 8, 13 -- Sat, 25 Nov 2006 08:14:45 -0600 8, 13 -- Mime-Version: 1.0 8, 13 -- Message-Id: {p06110401c18dff1f705c-at-[206.69.208.22]} 8, 13 -- In-Reply-To: {200611251408.kAPE8ME6032236-at-ns.microscopy.com} 8, 13 -- References: {200611251408.kAPE8ME6032236-at-ns.microscopy.com} 8, 13 -- Date: Sat, 25 Nov 2006 08:14:45 -0600 8, 13 -- To: maloneyb-at-fiu.edu, microscopy-at-microscopy.com 8, 13 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 8, 13 -- Subject: Re: viaWWW: Phillips 300 TEM 8, 13 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
I have accumulated a large number of colloidal gold conjugated antibodies and would like to determine if they are still good without having to screen each in some tedious immunocytochemical staining protocol. I assume one of two things can happen to inactivate an antibody-gold conjugate. The antibody could denature and make it ineffective. Alternatively, the antibody could disassociate from the colloidal gold particle and compete with bound antibody for the binding site which would result in reduced sensitivity. I blotted 50 ng (in 1 ul) of rabbit IgG onto nitrocellulose, blocked for several hours in 1% BSA and then incubated overnight at 4 C in 1:20 dilutions of my stock solutions. Some of my stocks dating back to 1998 still gave nice red dots after an overnight incubation. So at least some of the antibody conjugate is good. If I had been clever I would have tested each of these stocks against a dilution curve of antigen so I could evaluate whether there is reduced sensitivity. But I wasn't clever so now I am stuck trying to interpret which of these stocks are still usable. Does anyone know what level of antigen (i.e., target immunoglobulin) should be detectable by the various size gold particles (sensitivity in seeing blot bound colloidal gold must be linked to gold size). Is seeing a good spot at 50 ng/spot sufficient to proceed? In addition to my colloidal gold stocks, I tested 3 nanogold conjugates which were covalently linked. I naturally couldn't see then after the blot until I gold enhanced but since enhancement gave spots, they must be good since there can't be any dissociation - right? Is the minimally detectable concentration of IgG different for nanogold vs colloidal gold? Isn't this somewhat dependent on the enhancement step? Does anyone know if colloidal gold and/or nanogold conjugates can be kept at -20 C in 50% glycerol (my standard procedure for primary and fluorochrome conjugated antibodies?
I am interested in real world experience from everyone but the knowledgeable comments from Jan Leunissen and Rick Powell would be especially appreciated.
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 pleased and honored with your request to give you my comments and very much hope others might be willing to share their experiences. Colloid and adsorption physics are a very complicated business and not seldom have a lot of surprises.
In 1991 KRAMARCY and SEALOCK published a paper in JHC Vol. 39, No. 1, pp. 37-39, 1991: "Commercial Preparations of Colloidal Gold-Antibody Complexes Frequently Contain Free Active Antibody" Their data indicate that proteins adsorbed onto colloidal particles of 5nm and larger can dissociate from the particle surface with time and that, at times even shortly after manufacturing, colloidal gold reagents may contain free binding molecules. This is not necessarily the result of bad manufacturing practice as adsorption and desorption are in equilibrium at all times. Some proteins (there are even variations between antibodies from different animal species) are more liable to become dissociated than others and the conditions of coupling play a role as well. If dissociation does occur, than older conjugates will progressively loose activity as a result and this will be partly because of now less active reagent, and secondly because that reagent has now to compete with free (and thermodynamically more favorable) protein in solution.
Whether desorption will occur seems very much to be related to particle size. Ultra small conjugates prepared by adsorption have shown very consistent performance well beyond their shelf life. They do age well....As much as this may be a surprise, it is a pleasant one (at least for the user!) and I guess this illustrates that both adsorption based ultra small conjugates as well as antibodies can have a long and healthy life. Besides adsorption based production of gold reagents the covalent binding principle is used with small organo-metallic gold particles but I am not aware of this approach being used for larger particles.
How to test? What are sensible quality criteria? All producers of colloidal gold reagents (or any particle based immuno reagents for that matter) will have their own in house criteria, some may use additional enhancement even with large 'colored' particles, others may judge the performance exclusively based on the initial color.
So what does this mean in practice? With larger conjugates one should be able to easily detect spots "à vue" when they contain between 1 and 10 ng of specific protein in a dot-spot test under appropriate conditions that comply with the rules of affinity/avidity binding. Using enhancement the levels may go down into the picogram range. If you (or anyone else) should be interested, we have a newsletter available by Peter van de Plas describing in detail how to perform simple tests that demonstrate activity and performance of gold conjugates, primary antibodies and enhancement reagents, even down to the level of antigen recognition.
Storing conjugates....whenever one removes water from a hydrophilic structure that has partly hydrophobic qualities (as antibodies do) the risk of clustering based on hydrophobic interactions will definitely increase. And especially with conjugates built around large gold particles London/van der Waals interactions come into play as well. In case I would find myself at some point with a lot of conjugate left that was still reasonably 'fresh', I would store it at -20°C as a liquid (without freezing!) as you suggest; maybe rather use sucrose, as glycerol may affect membranes. I have no solid data for this but generally speaking desorption will be much reduced with lower temperatures, and should any clustering result from this treatment, a short spin of a ready-to-use diluted conjugate in a microfuge may remove such clusters.
Some of the above is based on our in-house or personal experience and may not necessarily have been publicly documented. Hope the info is helpful nevertheless. And finally: in spite of all the criteria I mentioned: sometimes old conjugates do remarkably well, even though their performance in a dot spot test would make one expect them to fail. I would say: if possible it is always worth a try with a well know test specimen and primary antibody combination.
Good luck!
Jan Leunissen
Aurion - President Electron Microscopy Research Advisor Costerweg 5 Dept Anatomy and Structural Biology 6702 A Wageningen Otago School of Medicine The Netherlands Dunedin, New Zealand phone 31-317-497676 phone 64-3-4795465 fax 31-317-415955 fax 64-3-4797254 http://www.aurion.nl http://anatomy.otago.ac.nz
==============================Original Headers============================== 16, 21 -- From leunissen-at-aurion.nl Sun Nov 26 19:42:07 2006 16, 21 -- Received: from fep04.xtra.co.nz (fep04.xtra.co.nz [210.54.141.242]) 16, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAR1g52q028898 16, 21 -- for {Microscopy-at-microscopy.com} ; Sun, 26 Nov 2006 19:42:06 -0600 16, 21 -- Received: from [192.168.1.50] (really [222.152.239.41]) by fep04.xtra.co.nz 16, 21 -- with ESMTP 16, 21 -- id {20061127014611.VYRD19621.fep04.xtra.co.nz-at-[192.168.1.50]} ; 16, 21 -- Mon, 27 Nov 2006 14:46:11 +1300 16, 21 -- In-Reply-To: {200611251919.kAPJJv2F029976-at-ns.microscopy.com} 16, 21 -- References: {200611251919.kAPJJv2F029976-at-ns.microscopy.com} 16, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 16, 21 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 16, 21 -- Message-Id: {0505EFBA-F4F2-463D-BF56-E58660DAF0BE-at-aurion.nl} 16, 21 -- Cc: Microscopy-at-microscopy.com 16, 21 -- From: Jan Leunissen {leunissen-at-aurion.nl} 16, 21 -- Subject: Re: [Microscopy] colloidal gold stability 16, 21 -- Date: Mon, 27 Nov 2006 14:46:10 +1300 16, 21 -- To: Tom Phillips {phillipst-at-missouri.edu} 16, 21 -- X-Mailer: Apple Mail (2.752.2) 16, 21 -- Content-Transfer-Encoding: 8bit 16, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kAR1g52q028898 ==============================End of - Headers==============================
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Email: neurowu-at-yahoo.com Name: mike
Organization: NDT
Title-Subject: [Filtered] price of used ccd camera
Question: Hi,
we are looking for a pre-owned ccd camera, such as optronics, Qimaging, Cooke, Roper, etc that can support our brightfield and/or fluorescent digital imagaing and quantitative analysis (volumentric analysis and cell, spine counting). Due to our budget limit, we can only offer less than USD1500. Please let me know if you have available one for sale or know the right party we can contact with, thanks!
Have you thought of an Astronomical camera? $1500 is tight, but it might cover an Artemis camera. I'd recommend one of the better Sony chips, e.g. ICX285AL or ICX285AQ. http://www.artemisccd.co.uk/icx285c.htm http://www.atik-instruments.com/
Good luck,
Austin
P.S. I'm not financially involved with Artemis; but I do have one of their excellent cameras.
----- Original Message ----- X-from: {neurowu-at-yahoo.com} To: {AuntDaisy-at-gmail.com} Sent: Monday, November 27, 2006 9:35 AM
Dear fellow microscopers,
Once again the efficiency of this list is breathtaking and I warmly thank all of you who spent some time explaining me the basics of SEM. I learned a lot in a short time and, though I had absolutely no experience in SEM, I can now run the microscope and even do EDX analysis (and EDX mapping! but this is thanks to the software). It goes without saying that I will need the help of an engineer or more experienced user to take the best of the instrument, but in the meantime I can get used to its manipulation.
Stephane
--- jmastrangelo-at-ultralifebatteries.com wrote:
} Hi Stephane, } } We purchased a TESCAN VEGAII last year, so I am by } no means an expert user } yet. Your problems with blurriness over 2Kx } magnification do sound familiar } to me, though. I was struggling with the same issues } when I was first } learning how to use the instrument. Have you had } training in the alignment / } beam optimization procedures yet? A few months after } the service tech } installed our instrument, TESCAN sent one of their } applications experts here } to our facilities for a couple days of training on } the instrument. He gave } me an alignment procedure that has greatly increased } the resolution in my } images. There is a lot of fine tuning to be done } with beam alignment and } with the stigmators to get good resolution at high } magnification. I do not } work at extremely high magnification but I have } acquired good images in the } 50-70Kx range. I have been able to resolve and image } particles of about } 20nm, although I believe the TESCAN people say the } instrument is good to } about 5nm for resolution. } } Hope this helps. If you like, I can send you the } general procedure I use to } optimize the imaging, although as I said- I am by no } means an expert user. } The guy at TESCAN that has been the greatest help to } me is Jack Mershon. He } could help you with more technical questions. Good } luck! } } Regards, } } ---------------------------------------- } Joe Mastrangelo } Chemical Lab Technician } Ultralife Batteries } 2000 Technology Parkway } Newark, NY 14513 } USA } } E-mail: jmastrangelo-at-ulbi.com } Phone: 315-359-6203 } Fax: 315-331-4264 } www.ultralifebatteries.com } } We. Are. Power. } } Want news, knowledge and commentary } from Ultralife experts? } Sign up for our Newsletter. } Subscribe at: www.ulbi.com/sign_up.php } } } } } } } -----Original Message----- } } From: nizets2-at-yahoo.com [SMTP:nizets2-at-yahoo.com] } } Sent: Thursday, November 23, 2006 10:48 AM } } To: jmastrangelo-at-ultralifebatteries.com } } Subject: [Microscopy] Re: Basic SEM functions } } } } } } } } } } } -------------------------------------------------------------------------- } } -- } } 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 finally found the necessary sequence to have } "green } } light" for "high vacuum" mode. Funnily, the vacuum } } remains the same, but now I have a green signal } and I } } can activate the HT and filament! } } Here are some few more questions: } } } } } } 1. In TEM one needs to tilt the sample to improve } the } } signal by EDX. Is it the same in SEM? (considering } } that the detector is also at an angle above the } } sample) } } 2. Is it realistic to use HT of 5kV or 10kV to } take } } EDX spectra by SEM? } } 3. Resolution issue: while observing mineral } } particules (quarz for example), when I reach a } } magnification of 2kx or higher (to observe } particles } } several µm big for example), the image is not } sharp. } } Using the mode which offers the maximum resolution } } (cleverly called "resolution mode"), the beam } reaches } } a size of 60nm. In these conditions it is not } } surprising that the image is not sharp. The SEM } allows } } magnification of up to 500kx!! What is the point } to } } use so high magnification if the image is blurred? } } What is the expected resolution of a SEM? } } } } Stephane } } } } } } } } } } } __________________________________________________________________________ } } __________ } } Do you Yahoo!? } } Everyone is raving about the all-new Yahoo! Mail } beta. } } http://new.mail.yahoo.com } } } } ==============================Original } } Headers============================== } } 7, 19 -- From nizets2-at-yahoo.com Thu Nov 23 } 09:39:03 2006 } } 7, 19 -- Received: from } web37415.mail.mud.yahoo.com } } (web37415.mail.mud.yahoo.com [209.191.91.147]) } } 7, 19 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } } kANFd2jH020698 } } 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 23 } Nov 2006 09:39:03 } } -0600 } } 7, 19 -- Received: (qmail 87141 invoked by uid } 60001); 23 Nov 2006 } } 15:39:02 -0000 } } 7, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; } c=nofws; } } 7, 19 -- s=s1024; d=yahoo.com; } } 7, 19 -- } } } h=X-YMail-OSG:Received:Date:From:Subject:To:MIME-Version:Content-Type:Cont } } ent-Transfer-Encoding:Message-ID; } } 7, 19 -- } } } b=hVWsNBLdPT8OJ+cRYh7ebPum4IIFJrBmR3iGwec0wlKHA9DoHA/QWcz0Ny0TO9FKzCQRrE12 } } } i3MJSOdXHJM4OVfZ4rYzpf0z9rcmRh/NyJ3UWHQk9oIbGFPSeiECDSFAEmz9nmscWmfDHnv7Nm } } VV0umhPc2YuFBrC2FUoLNlOJE=; } } 7, 19 -- X-YMail-OSG: } } } XhFNP5wVM1mltPNRJpdw3q2cRh2TsqpyvuUNWwrM4nNJqo0e9k0iew.rE6kvTC5T8Ni3NYra.t } } } cG2iUuqjZe7gs1DNcPR43b1HKoYNNKJeY93sCBEktlIJhBjuNmsOCDwjsJ2PAXT_qUFfM- } } 7, 19 -- Received: from [80.122.101.102] by } web37415.mail.mud.yahoo.com } } via HTTP; Thu, 23 Nov 2006 07:39:02 PST } } 7, 19 -- Date: Thu, 23 Nov 2006 07:39:02 -0800 } (PST) } } 7, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} } } 7, 19 -- Subject: Re: Basic SEM functions } } 7, 19 -- To: microscopy-at-microscopy.com } } 7, 19 -- MIME-Version: 1.0 } } 7, 19 -- Content-Type: text/plain; } charset=iso-8859-1 } } 7, 19 -- Content-Transfer-Encoding: 8bit } } 7, 19 -- Message-ID: } {292322.86779.qm-at-web37415.mail.mud.yahoo.com} } } ==============================End of - } } Headers============================== } } This email was scanned for viruses before leaving } the Ultralife Batteries, Inc. network. }
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==============================Original Headers============================== 11, 20 -- From nizets2-at-yahoo.com Mon Nov 27 09:34:48 2006 11, 20 -- Received: from web37407.mail.mud.yahoo.com (web37407.mail.mud.yahoo.com [209.191.91.139]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kARFYl1P016224 11, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Nov 2006 09:34:47 -0600 11, 20 -- Received: (qmail 87231 invoked by uid 60001); 27 Nov 2006 15:34:46 -0000 11, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 20 -- s=s1024; d=yahoo.com; 11, 20 -- h=X-YMail-OSG:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 11, 20 -- b=voxU3m4hEY08OVlwP6we+6HuVt7zCnElMbwksO2zAT0f+eBi+DTbSBVoXHzEHxg09pexxmoDKS+p9qkdcwsOMFFYcgMR2fzg/Nn34zRZzWG+4bpRQhKJoqbeQVxi+wu/azuKU7CDpHvGf5G+G+smkhpmiTMvU7arK8zXV3CgYB8=; 11, 20 -- X-YMail-OSG: 3ZZwG1cVM1lW482zaEfijpCY7KNryRqOGtikNSdlWdDPj76B0avT0EVO9Ic7Lwr9wKx9u.2eGWAVY5LMsgcSyqNgeVvIez_aIQzOWih_9s2850AnHXHckOLaB.gi3MhVu.PIxOJyXgrcJAEr5IJd94b35Y4JHT4QBoIqQQpVgICDBbXEyWTeddKuc_Zi2PZtmyG405Af 11, 20 -- Received: from [80.122.101.102] by web37407.mail.mud.yahoo.com via HTTP; Mon, 27 Nov 2006 07:34:46 PST 11, 20 -- Date: Mon, 27 Nov 2006 07:34:46 -0800 (PST) 11, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 11, 20 -- Subject: RE: [Microscopy] Re: Basic SEM functions - thank you 11, 20 -- To: microscopy-at-microscopy.com 11, 20 -- In-Reply-To: {A0C02A449BCBD311AEA10008C75C2A6C0856C1EB-at-PM_GENERAL} 11, 20 -- MIME-Version: 1.0 11, 20 -- Content-Type: text/plain; charset=iso-8859-1 11, 20 -- Content-Transfer-Encoding: 8bit 11, 20 -- Message-ID: {629664.87163.qm-at-web37407.mail.mud.yahoo.com} ==============================End of - Headers==============================
I am sorry if my email behavior caused anybody any discomfort. I don't understand why it is so, I don't understand what I do wrong and I cannot change it(sorry I am no email expert). All I can propose is to stop emailing me if you think that I do something wrong.
Stephane
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==============================Original Headers============================== 4, 20 -- From nizets2-at-yahoo.com Mon Nov 27 09:43:56 2006 4, 20 -- Received: from web37405.mail.mud.yahoo.com (web37405.mail.mud.yahoo.com [209.191.91.137]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kARFhtFw027246 4, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Nov 2006 09:43:55 -0600 4, 20 -- Received: (qmail 37439 invoked by uid 60001); 27 Nov 2006 15:43:54 -0000 4, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 20 -- s=s1024; d=yahoo.com; 4, 20 -- h=X-YMail-OSG:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 4, 20 -- b=YuTFgvVAKgVH35/oOyFd1d83gVCYmpo5ES3B7lHZWqZhc5n/2TGGcHINOHeexesYlXrd31vAlGBmm/cXlc+V8UsGerEC/wM95Dbc7Tw5v+LpFJqL3OEJ40NGJc3j5CU/7YKa1w3bUFi0h6uALmUAw2M/u2WXdEApos5vd22p7Uo=; 4, 20 -- X-YMail-OSG: C1fKULEVM1kJ1ua5E47_75D8cGaWdcCXahIQREc2xGShe1.ptbPi6shtQriNq_lYmpfV8otvMpvHFYqxo7gmeExlaSBA7KHdaPc2xWPrjH2Ccg90iicrf2p.isKYrW0rT0tRxhHqH16PAw-- 4, 20 -- Received: from [80.122.101.102] by web37405.mail.mud.yahoo.com via HTTP; Mon, 27 Nov 2006 07:43:54 PST 4, 20 -- Date: Mon, 27 Nov 2006 07:43:54 -0800 (PST) 4, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 4, 20 -- Subject: Re: [Microscopy] Re: Basic SEM functions 4, 20 -- To: microscopy-at-microscopy.com 4, 20 -- In-Reply-To: {200611252217.kAPMHOt29404-at-www.matscieng.sunysb.edu} 4, 20 -- MIME-Version: 1.0 4, 20 -- Content-Type: text/plain; charset=iso-8859-1 4, 20 -- Content-Transfer-Encoding: 8bit 4, 20 -- Message-ID: {688158.37253.qm-at-web37405.mail.mud.yahoo.com} ==============================End of - Headers==============================
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Email: rpowell-at-nanoprobes.com Name: Rick Powell
I'm also honored to be asked...our experience with conventional colloidal gold is a little less than Jan's, but from our experience, your test is a good one for conjugates of 5, 10 and 15 nm colloidal gold, where 50 ng of target gives a reasonably clear red spot (we obtain progressively lighter spots down to about 5 or 10 ng, sometimes 1 ng for 15 nm gold, for good preps); the spot will be darker for larger colloidal gold particles. A good deep red spot would be a good indication of a working conjugate.
With NanogoldÆ, for product testing purposes, we consider clear detection of 10 ng of target after silver enhancement to be acceptable (In practice, we can usually detect 0.1, and frequently 0.01 ng of target after silver enhancement). 50 ng of target is a little more than I would have chosen as a test amount - i.e. it may be enough to give you a signal even if the antibody has lost some native affinity (I would suggest 1 or 5 ng). However, as with colloidal gold, the spots become progressively lighter with smaller amounts of target, and if you compare your blots with one that shows how the spots look for a series of dilutions of target, it will give you a good qualitative idea of how good your conjugates are.
Our web site has a photo of a blot where we used Nanogold-labeled Fab' anti-mouse to detect serial dilutions of mouse IgG:
http://www.nanoprobes.com/NanoAb.html#blot
The first (darkest) pair of spots (top left) contain 10 ng of target IgG, reading left to right along the top row, the second pair contains 5 ng, and the third 1 ng; if your 50 ng spots look like any of these three after you develop them with gold enhancement, the conjugates are definitely still good. After silver or gold enhancement, the spots should be black (sometimes purplish, sometimes brownish) rather than deep red.
More images are available in Hainfeld and Furuya's paper describing Nanogold (Hainfeld, J.F. and Furuya, F.R. A 1.4nm Gold cluster covalently attached to antibodies improves immunolabeling, J. Histochem. Cytochem., 40, 177-184 (1992)); PDF reprint available at http://www.jhc.org/cgi/reprint/40/2/177.
We don't usually freeze Nanogold conjugates, but freezing at -20ƒC in 30% glycerol is usually recommended to avoid ice crystal damage, and your procedure with 50% glycerol should be equally effective. Provided they are not allowed to dry out or become contaminated, Nanogold conjugates can last well beyond the year's shelf life if they are refrigerated at 4ƒC; we have tested them after two or three years and found blot detection sensitivity to be unchanged (we include a bacterial inhibitor).
Hope this helps,
Rick Powell Nanoprobes, Incorporated www.nanoprobes.com
I have accumulated a large number of colloidal gold conjugated antibodies and would like to determine if they are still good without having to screen each in some tedious immunocytochemical staining protocol. I assume one of two things can happen to inactivate an antibody-gold conjugate. The antibody could denature and make it ineffective. Alternatively, the antibody could disassociate from the colloidal gold particle and compete with bound antibody for the binding site which would result in reduced sensitivity. I blotted 50 ng (in 1 ul) of rabbit IgG onto nitrocellulose, blocked for several hours in 1% BSA and then incubated overnight at 4 C in 1:20 dilutions of my stock solutions. Some of my stocks dating back to 1998 still gave nice red dots after an overnight incubation. So at least some of the antibody conjugate is good. If I had been clever I would have tested each of these stocks against a dilution curve of antigen so I could evaluate whether there is reduced sensitivity. But I wasn't clever so now I am stuck trying to interpret which of these stocks are still usable. Does anyone know what level of antigen (i.e., target immunoglobulin) should be detectable by the various size gold particles (sensitivity in seeing blot bound colloidal gold must be linked to gold size). Is seeing a good spot at 50 ng/spot sufficient to proceed? In addition to my colloidal gold stocks, I tested 3 nanogold conjugates which were covalently linked. I naturally couldn't see then after the blot until I gold enhanced but since enhancement gave spots, they must be good since there can't be any dissociation - right? Is the minimally detectable concentration of IgG different for nanogold vs colloidal gold? Isn't this somewhat dependent on the enhancement step? Does anyone know if colloidal gold and/or nanogold conjugates can be kept at -20 C in 50% glycerol (my standard procedure for primary and fluorochrome conjugated antibodies?
I am interested in real world experience from everyone but the knowledgeable comments from Jan Leunissen and Rick Powell would be especially appreciated.
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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==============================Original Headers============================== 15, 20 -- From MCarlyle-at-veeco.com Mon Nov 27 13:26:34 2006 15, 20 -- Received: from sboexch2.int.veeco.com (SBOEXCH2.VEECO.com [68.111.35.167]) 15, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kARJQXj2022562 15, 20 -- for {Microscopy-at-Microscopy.Com} ; Mon, 27 Nov 2006 13:26:33 -0600 15, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 15, 20 -- content-class: urn:content-classes:message 15, 20 -- MIME-Version: 1.0 15, 20 -- Content-Type: text/plain; 15, 20 -- charset="iso-8859-1" 15, 20 -- Subject: AFM -- Seeing at the Nanoscale V Conference Call for Papers 15, 20 -- Date: Mon, 27 Nov 2006 11:26:32 -0800 15, 20 -- Message-ID: {625BD6DD96DE554DBB0558FFE70E0F4201AD58B2-at-sboexch2.int.veeco.com} 15, 20 -- X-MS-Has-Attach: 15, 20 -- X-MS-TNEF-Correlator: 15, 20 -- Thread-Topic: AFM -- Seeing at the Nanoscale V Conference Call for Papers 15, 20 -- Thread-Index: AccSWfHhVuXmyKsVTfiPxBebYfBLrw== 15, 20 -- From: "Marlene Carlyle" {MCarlyle-at-veeco.com} 15, 20 -- To: {Microscopy-at-Microscopy.Com} 15, 20 -- Content-Transfer-Encoding: 8bit 15, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kARJQXj2022562 ==============================End of - Headers==============================
Sometimes I want to get rid of mucous, but sometimes I want to preserve it. In this case the researchers want to look at microbes in the mucous surrounding corals, as well as microbes within the coral tissue with TEM. I have had pretty good success decalcifying corals for TEM, but it's been a long time since I wanted to keep slime. I know people use ruthenium red and Alcian blue to stain mucous, but do these preserve it throughout processing for TEM as well? I also have some ruthenium tetroxide in the fridge from the early 1990s; I think it was once used for preserving slimy biofilms.
Any suggestions welcomed!
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============================== 6, 19 -- From tina-at-pbrc.hawaii.edu Mon Nov 27 19:35:11 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 kAS1ZA94010133 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Nov 2006 19:35:10 -0600 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 kAS1Z6x4011562 6, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Nov 2006 15:35:06 -1000 (HST) 6, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id kAS1Z5B2011559 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Nov 2006 15:35:05 -1000 (HST) 6, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 6, 19 -- Date: Mon, 27 Nov 2006 15:35:04 -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: Preservation of microbes in external mucous 6, 19 -- Message-ID: {Pine.GSO.4.21.0611271530510.11499-100000-at-halia} 6, 19 -- MIME-Version: 1.0 6, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
I am looking for a database for atomic positions (Wyckoff Positions ) for compounds (metals, oxides and ceramics) and software to construct structures to view planes, different sites (octahedral, tetrahedral, etc.), and diffraction simulation. Also any recommendations for a good book in practical electron microscopy which includes the new microscopy techniques. I would appreciate any information on the above issues. Thank you,
Can anyone point me to the specification of the Cambridge S90 SEM and when the machine was first marketed? I tried google but nothing relevant came uo. 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============================== 2, 20 -- From wpchan-at-u.washington.edu Mon Nov 27 22:54:41 2006 2, 20 -- Received: from mxout5.cac.washington.edu (mxout5.cac.washington.edu [140.142.32.135]) 2, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAS4sfku003095 2, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Nov 2006 22:54:41 -0600 2, 20 -- Received: from homer22.u.washington.edu (homer22.u.washington.edu [140.142.15.9]) 2, 20 -- by mxout5.cac.washington.edu (8.13.7+UW06.06/8.13.7+UW06.09) with ESMTP id kAS4se9L018878 2, 20 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 2, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Nov 2006 20:54:40 -0800 2, 20 -- Received: from localhost (wpchan-at-localhost) 2, 20 -- by homer22.u.washington.edu (8.13.7+UW06.06/8.13.7+Submit) with ESMTP id kAS4seWC032644 2, 20 -- for {microscopy-at-microscopy.com} ; Mon, 27 Nov 2006 20:54:40 -0800 2, 20 -- Date: Mon, 27 Nov 2006 20:54:40 -0800 (PST) 2, 20 -- From: "W. Chan" {wpchan-at-u.washington.edu} 2, 20 -- To: microscopy-at-microscopy.com 2, 20 -- Subject: cambridge S90 2, 20 -- Message-ID: {Pine.LNX.4.64.0611272015270.28974-at-homer22.u.washington.edu} 2, 20 -- MIME-Version: 1.0 2, 20 -- Content-Type: TEXT/PLAIN; charset=US-ASCII; format=flowed 2, 20 -- X-PMX-Version: 5.2.2.285561, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2006.11.27.204432 2, 20 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
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Email: brunnerrobert-at-netzero.com Name: Robert Brunner
Organization: Surpass Inc.
Title-Subject: [Filtered] Tissue Prep and hold for TEM and or SEM
Question: I am working with a Scientist that wants to collect tissue from a study animal to hold for potential SEM and or TEM. I was wondering if anyone can provide any procedures for preparation of the tissue from collection to fixation to storage that would give optimal results for future SEM and TEM studies. It is not known how long the samples will be stored so please let me know if there is a time limit on the storage of the samples also.
I have found the ICSD database useful. You may have to subscribe first. The database is pretty complete and I've found it quite useful. See:
http://icsdweb.fiz-karlsruhe.de/index.php
The other place to look is Pearson's Handbook of Crystallographic Data for Intermetallic Phases(edited by Villars and Calvert), which is a particularly good reference for metals but is only partial for oxides.
Regards, Wharton
-----Original Message----- X-from: jafarhan-at-rci.rutgers.edu [mailto:jafarhan-at-rci.rutgers.edu] Sent: Monday, November 27, 2006 8:15 PM To: Sinkler, Wharton
Dear folks,
I am looking for a database for atomic positions (Wyckoff Positions ) for compounds (metals, oxides and ceramics) and software to construct structures to view planes, different sites (octahedral, tetrahedral, etc.), and diffraction simulation. Also any recommendations for a good book in practical electron microscopy which includes the new microscopy techniques. I would appreciate any information on the above issues. Thank you,
What tissue? What type of study? Can the tissues be freeze-fixed? I collected and processed tissues in Antarctica for study back in Chicago, but my methods may not be relevant to the studies. And given that one of the trips was done in the days of Pel-Dri, before it became illegal, that method is definitely not relevant. Hm. How does the tissue work with tert-butyl alcohol? Might try dehydrating with that, and then letting the tBA freeze (pretty much freezes at room temp.), and storing that way. I've never tried this and have no idea if it would work, but tBA has been used for light microscopy, and as a dehydrating/freeze-sublimation drying method for SEM (there's a reference for this, but I haven't found it in my horizontal filiing system). If the tissues can be freeze-fixed, a possible method would be EM freeze-fixation and storage well below the recrystallization temperature (I'd use -90 C or better) then careful rewarming and freeze-substitution. But again, I haven't tried that for storage.
Phil
} 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 brunnerrobert-at-netzero.com as well as the } MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: brunnerrobert-at-netzero.com } Name: Robert Brunner } } Organization: Surpass Inc. } } Title-Subject: [Filtered] Tissue Prep and hold for TEM and or SEM } } Question: I am working with a Scientist that wants to collect tissue } from a study animal to hold for potential SEM and or TEM. I was } wondering if anyone can provide any procedures for preparation of the } tissue from collection to fixation to storage that would give optimal } results for future SEM and TEM studies. It is not known how long the } samples will be stored so please let me know if there is a time limit } on the storage of the samples also. } } Thanks for any help } } } Robert Brunner -- Philip Oshel Microscopy Facility Supervisor Biology Department Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 voice: (989) 774-3576 dept. fax: (989) 774-3462
==============================Original Headers============================== 3, 23 -- From oshel1pe-at-cmich.edu Tue Nov 28 07:52:35 2006 3, 23 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kASDqZaD014715 3, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 07:52:35 -0600 3, 23 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 23 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id kASEM2N4004123; 3, 23 -- Tue, 28 Nov 2006 09:22:09 -0500 3, 23 -- Received: from [141.209.160.249] ([141.209.160.249]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 23 -- Tue, 28 Nov 2006 08:52:25 -0500 3, 23 -- Mime-Version: 1.0 3, 23 -- Message-Id: {f06230918c191eab08cbb-at-[141.209.160.249]} 3, 23 -- In-Reply-To: {200611280802.kAS82uiZ024523-at-ns.microscopy.com} 3, 23 -- References: {200611280802.kAS82uiZ024523-at-ns.microscopy.com} 3, 23 -- Date: Tue, 28 Nov 2006 08:52:24 -0500 3, 23 -- To: brunnerrobert-at-netzero.com 3, 23 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 23 -- Subject: Re: [Microscopy] viaWWW: Tissue Prep and hold for TEM and or SEM 3, 23 -- Cc: Microscopy-at-microscopy.com 3, 23 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 23 -- X-OriginalArrivalTime: 28 Nov 2006 13:52:25.0790 (UTC) FILETIME=[6FA73DE0:01C712F4] 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==============================
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 catherine.bougerol-at-cea.fr as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] preparation of nanowires for TEM
Question: I have got ZnSe nanowires grown on a GaAs substrate. Up to now, I have prepared them by putting a piece of the sample in methanol and letting it for one hour in an ultrasonic device ; them I put a drop of the solution on a copper carbon grid. With this method, the nanowires are broken and one cannot study the interface with the substrate. This is why I am trying to find another method of preparation keeping them on the substrate.
Besides the things you mentioned, there are techniques for "non-aqueous" fixation to preserve biofilms using fluorocarbon solvents, such as Fluorinert FC-72, which is manufactured by the 3M company. Call them for distributor information. Also high-pressure freezing and freeze-substitution are options, and dehydration in HMDS can help retain mucins for SEM studies.
References to check:
P. Allan-Wojtas et al., Micros. Res. Tech., 36:390-399 (1997)
Sims, DE et al. Biotech Histochem. 66(4):173-80 (1991)
Sanchez et al. J Comp Path. 117(2):165-70 (1997)
Sims and Horne Am J Physiol 273: L1036-L1041 (1997)
Bock et al. Biotech Histochem 74(5):244-7 (1999)
There....that ought to get you started. Hope it helps!
65 and muggy in Columbia.
Cheers, Randy
Randy Tindall Senior EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu On-line calendar: http://biotech.rnet.missouri.edu/cgi-bin/Calcium310.pl?Op=Splash&Amount= Week&NavType=Both&Type=TimePlan
-----Original Message----- X-from: tina-at-pbrc.hawaii.edu [mailto:tina-at-pbrc.hawaii.edu] Sent: Monday, November 27, 2006 7:37 PM To: Tindall, Randy D.
Hi, All-
Sometimes I want to get rid of mucous, but sometimes I want to preserve it. In this case the researchers want to look at microbes in the mucous surrounding corals, as well as microbes within the coral tissue with TEM. I have had pretty good success decalcifying corals for TEM, but it's been a long time since I wanted to keep slime. I know people use ruthenium red and Alcian blue to stain mucous, but do these preserve it throughout processing for TEM as well? I also have some ruthenium tetroxide in the fridge from the early 1990s; I think it was once used for preserving slimy biofilms.
Any suggestions welcomed!
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============================== 6, 19 -- From tina-at-pbrc.hawaii.edu Mon Nov 27 19:35:11 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 kAS1ZA94010133 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Nov 2006 19:35:10 -0600 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 kAS1Z6x4011562 6, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Nov 2006 15:35:06 -1000 (HST) 6, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id kAS1Z5B2011559 6, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 27 Nov 2006 15:35:05 -1000 (HST) 6, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 6, 19 -- Date: Mon, 27 Nov 2006 15:35:04 -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: Preservation of microbes in external mucous 6, 19 -- Message-ID: {Pine.GSO.4.21.0611271530510.11499-100000-at-halia} 6, 19 -- MIME-Version: 1.0 6, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
==============================Original Headers============================== 27, 25 -- From TindallR-at-missouri.edu Tue Nov 28 08:54:49 2006 27, 25 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 27, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kASEsnKW005045 27, 25 -- for {microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 08:54:49 -0600 27, 25 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 27, 25 -- Tue, 28 Nov 2006 08:54:48 -0600 27, 25 -- x-mimeole: Produced By Microsoft Exchange V6.5 27, 25 -- Content-class: urn:content-classes:message 27, 25 -- MIME-Version: 1.0 27, 25 -- Content-Type: text/plain; 27, 25 -- charset="us-ascii" 27, 25 -- Subject: RE: [Microscopy] Preservation of microbes in external mucous 27, 25 -- Date: Tue, 28 Nov 2006 08:54:48 -0600 27, 25 -- Message-ID: {91108EF9255B394CBF8B7E3789814A41C68C03-at-UM-XMAIL08.um.umsystem.edu} 27, 25 -- In-Reply-To: {200611280136.kAS1aqD6012158-at-ns.microscopy.com} 27, 25 -- X-MS-Has-Attach: 27, 25 -- X-MS-TNEF-Correlator: 27, 25 -- Thread-Topic: [Microscopy] Preservation of microbes in external mucous 27, 25 -- Thread-Index: AccSlDWQ1XmlUiCcSPOIQrO5ufSEGwAZIn2Q 27, 25 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 27, 25 -- To: {tina-at-pbrc.hawaii.edu} 27, 25 -- Cc: {microscopy-at-microscopy.com} 27, 25 -- X-OriginalArrivalTime: 28 Nov 2006 14:54:48.0039 (UTC) FILETIME=[26351F70:01C712FD] 27, 25 -- Content-Transfer-Encoding: 8bit 27, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kASEsnKW005045 ==============================End of - Headers==============================
A group in the Geology Department at Arizona State University maintains an internet site for the American Mineralogist Crystal Structure Database. It's free, contains atomic position data for most minerals and has useful information not in Pearson's Handbook. The address is rruff.geo.arizona.edu/AMS/amcsd.php.
Larry Thomas PNNL
On 11/28/06 5:23 AM, "Wharton.Sinkler-at-uop.com" {Wharton.Sinkler-at-uop.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 Jafar, } } I have found the ICSD database useful. You may have to subscribe first. } The database is pretty complete and I've found it quite useful. See: } } http://icsdweb.fiz-karlsruhe.de/index.php } } The other place to look is Pearson's Handbook of Crystallographic Data } for Intermetallic Phases(edited by Villars and Calvert), which is a } particularly good reference for metals but is only partial for oxides. } } Regards, } Wharton } } -----Original Message----- } X-from: jafarhan-at-rci.rutgers.edu [mailto:jafarhan-at-rci.rutgers.edu] } Sent: Monday, November 27, 2006 8:15 PM } To: Sinkler, Wharton } Subject: [Microscopy] Atomic positions database and software } } } } } ------------------------------------------------------------------------ } ---- } 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 folks, } } I am looking for a database for atomic positions (Wyckoff Positions ) } for } compounds (metals, oxides and ceramics) and software to construct } structures to view planes, different sites (octahedral, tetrahedral, } etc.), and diffraction simulation. Also any recommendations for a good } book in practical electron microscopy which includes the new microscopy } techniques. } I would appreciate any information on the above issues. } Thank you, } } Jafar } } Materials Science and Engineering } Rutgers University, } Piscataway, NJ 08854 } Tel. 732 445 5615 } } ==============================Original } Headers============================== } 5, 21 -- From jafarhan-at-rci.rutgers.edu Mon Nov 27 20:13:28 2006 } 5, 21 -- Received: from rci.rutgers.edu (rci-out.rutgers.edu } [128.6.68.135]) } 5, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id kAS2DSdR021489 } 5, 21 -- for {microscopy-at-microscopy.com} ; Mon, 27 Nov 2006 } 20:13:28 -0600 } 5, 21 -- Received: by rci.rutgers.edu (Postfix, from userid 11335) } 5, 21 -- id DE1BA12D6; Mon, 27 Nov 2006 21:13:27 -0500 (EST) } 5, 21 -- Received: from 68.192.253.101 } 5, 21 -- (SquirrelMail authenticated user jafarhan) } 5, 21 -- by webmail.rci.rutgers.edu with HTTP; } 5, 21 -- Mon, 27 Nov 2006 21:13:27 -0500 (EST) } 5, 21 -- Message-ID: } {3589.68.192.253.101.1164680007.squirrel-at-webmail.rci.rutgers.edu} } 5, 21 -- Date: Mon, 27 Nov 2006 21:13:27 -0500 (EST) } 5, 21 -- Subject: Atomic positions database and software } 5, 21 -- From: jafarhan-at-rci.rutgers.edu } 5, 21 -- To: microscopy-at-microscopy.com } 5, 21 -- User-Agent: SquirrelMail/1.4.7 } 5, 21 -- MIME-Version: 1.0 } 5, 21 -- Content-Type: text/plain;charset=iso-8859-1 } 5, 21 -- Content-Transfer-Encoding: 8bit } 5, 21 -- X-Priority: 3 (Normal) } 5, 21 -- Importance: Normal } ==============================End of - } Headers============================== } } } ==============================Original Headers============================== } 16, 27 -- From Wharton.Sinkler-at-uop.com Tue Nov 28 07:18:21 2006 } 16, 27 -- Received: from AZ18CN849.global.ds.honeywell.com } (tmpnat1.honeywell.com [199.64.0.252]) } 16, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } kASDIKln003069 } 16, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 07:18:21 -0600 } 16, 27 -- Received: from dplexch7.uop.com ([138.90.2.112]) by } AZ18CN849.global.ds.honeywell.com with Microsoft SMTPSVC(6.0.3790.1830); } 16, 27 -- Tue, 28 Nov 2006 06:18:20 -0700 } 16, 27 -- Received: from DPLEVS1.ad.uop.com ([138.90.2.100]) by } dplexch7.uop.com with Microsoft SMTPSVC(6.0.3790.1830); } 16, 27 -- Tue, 28 Nov 2006 07:18:19 -0600 } 16, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 16, 27 -- Content-class: urn:content-classes:message } 16, 27 -- MIME-Version: 1.0 } 16, 27 -- Content-Type: text/plain; } 16, 27 -- charset="us-ascii" } 16, 27 -- Subject: RE: [Microscopy] Atomic positions database and software } 16, 27 -- Date: Tue, 28 Nov 2006 07:19:00 -0600 } 16, 27 -- Message-ID: } {62C8A7975D4AE545A23A0CD278BB20A907DFB975-at-DPLEVS1.ad.uop.com} } 16, 27 -- In-Reply-To: {200611280215.kAS2FG1s023788-at-ns.microscopy.com} } 16, 27 -- X-MS-Has-Attach: } 16, 27 -- X-MS-TNEF-Correlator: } 16, 27 -- Thread-Topic: [Microscopy] Atomic positions database and software } 16, 27 -- Thread-Index: AccSkxDEYrvna2kATX2F0pAiPeAQBwAW/BHw } 16, 27 -- From: "Sinkler, Wharton" {Wharton.Sinkler-at-uop.com} } 16, 27 -- To: {jafarhan-at-rci.rutgers.edu} } 16, 27 -- Cc: {microscopy-at-microscopy.com} } 16, 27 -- X-OriginalArrivalTime: 28 Nov 2006 13:18:19.0919 (UTC) } FILETIME=[AC3819F0:01C712EF] } 16, 27 -- Content-Transfer-Encoding: 8bit } 16, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id kASDIKln003069 } ==============================End of - Headers==============================
==============================Original Headers============================== 6, 26 -- From Larry.Thomas-at-pnl.gov Tue Nov 28 10:44:26 2006 6, 26 -- Received: from emailgw01.pnl.gov (emailgw01.pnl.gov [192.101.109.33]) 6, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kASGiPgB019046 6, 26 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 10:44:26 -0600 6, 26 -- Received: from pnlmse21.pnl.gov ([130.20.128.55]) 6, 26 -- by emailgw01.pnl.gov with ESMTP; 28 Nov 2006 08:44:25 -0800 6, 26 -- X-IronPort-AV: i="4.09,470,1157353200"; 6, 26 -- d="scan'208"; a="12730052:sNHT31707347" 6, 26 -- Received: from EMAIL02.pnl.gov ([130.20.128.221]) by pnlmse21.pnl.gov with Microsoft SMTPSVC(6.0.3790.1830); 6, 26 -- Tue, 28 Nov 2006 08:44:25 -0800 6, 26 -- Received: from 130.20.26.238 ([130.20.26.238]) by EMAIL02.pnl.gov ([130.20.128.155]) via Exchange Front-End Server webmaili.pnl.gov ([130.20.248.53]) with Microsoft Exchange Server HTTP-DAV ; 6, 26 -- Tue, 28 Nov 2006 16:44:25 +0000 6, 26 -- User-Agent: Microsoft-Entourage/11.2.4.060510 6, 26 -- Date: Tue, 28 Nov 2006 08:44:24 -0800 6, 26 -- Subject: [Microscopy] RE: Atomic positions database and software 6, 26 -- From: "Thomas, Larry (PNNL)" {Larry.Thomas-at-pnl.gov} 6, 26 -- To: {Microscopy-at-microscopy.com} 6, 26 -- Message-ID: {C191A768.16DF%Larry.Thomas-at-pnl.gov} 6, 26 -- Thread-Topic: [Microscopy] RE: Atomic positions database and software 6, 26 -- Thread-Index: AccTDHXJtF8OZH7/EduQ8QAWyzlS+g== 6, 26 -- In-Reply-To: {200611281323.kASDNfSK008566-at-ns.microscopy.com} 6, 26 -- Mime-version: 1.0 6, 26 -- Content-type: text/plain; 6, 26 -- charset="US-ASCII" 6, 26 -- Content-transfer-encoding: 7bit 6, 26 -- X-OriginalArrivalTime: 28 Nov 2006 16:44:25.0222 (UTC) FILETIME=[76839A60:01C7130C] ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (blegge-at-cwisp.ca) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, November 28, 2006 at 12:21:31 ---------------------------------------------------------------------------
Email: blegge-at-cwisp.ca Name: Brian Legge
Education: 9-12th Grade High School
Location: Canada
Question: Hello I am looking for a source for OM sample slides. A school in Cyprus needs to upgrade their science studies facilities and slides are needed.They are using a British based curriculum. Human tissue- nerve, spinal chord,sperm and other mammal tissues etc as well as botany -roots, stems, leaves etc. Where do your school boards obtain slides for grades 9-12? Thank you
Hello All FUNDATEL Foundation, a non profit organization (www.fundatel.org.ar) , is looking for used ( and working ) TEM, SEM and SPM ( and related equipment ) to received it in DONATION. This equipment will be used in our R+D proyects in the biosensors and MEM´s area. Please contact us as soon as posible, we response inmediatly and pay all the costs of shipping and handling.
Best Regards
-- Fernando D. Balducci President FUNDATEL Foundation Parana - Entre Rios Argentina
email: fernando.balducci-at-fundatel.org.ar
==============================Original Headers============================== 7, 23 -- From fundatel-at-gmail.com Tue Nov 28 13:52:54 2006 7, 23 -- Received: from wx-out-0506.google.com (wx-out-0506.google.com [66.249.82.233]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kASJqr1o013219 7, 23 -- for {microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 13:52:54 -0600 7, 23 -- Received: by wx-out-0506.google.com with SMTP id h30so2249292wxd 7, 23 -- for {microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 11:52:52 -0800 (PST) 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=KyQO+kjrPiThHX3h73W9mnjo9NvjMyzojiCGBl3jG2HPUkGoX1PfyeYkiDA1MVR1JXW/54Fbs45mOEE5yuhRUHzXAyHlG4Edu8ZPz5ME5wTPmtPkUj3bISTyS9E3cltRzPxJB/QuSlhaEN5fFIuqeH5eVRPO2+GYiHI6xKmbGQg= 7, 23 -- Received: by 10.70.44.4 with SMTP id r4mr2353797wxr.1164743572268; 7, 23 -- Tue, 28 Nov 2006 11:52:52 -0800 (PST) 7, 23 -- Received: by 10.70.62.6 with HTTP; Tue, 28 Nov 2006 11:52:51 -0800 (PST) 7, 23 -- Message-ID: {715613900611281152n234253cma16cfdfcf8827a4a-at-mail.gmail.com} 7, 23 -- Date: Tue, 28 Nov 2006 16:52:51 -0300 7, 23 -- From: Fundatel {fundatel-at-gmail.com} 7, 23 -- To: microscopy-at-microscopy.com 7, 23 -- Subject: looking for SEM and TEM 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 kASJqr1o013219 ==============================End of - Headers==============================
Has anyone yet tried the newish Microsoft VX-6000 Webcam? It claims to give 5MP for stills, it's cheap, I'm tempted to get one, rip the lens off, and patch it onto the top of a microscope with sticky tape. Not much to lose.
cheers
rtch
O} } Dear Mike, } } Do you want colour or B&W + filters? } } Have you thought of an Astronomical camera? $1500 is tight, but } it } might cover an Artemis camera. I'd recommend one of the better Sony } chips, e.g. ICX285AL or ICX285AQ. } http://www.artemisccd.co.uk/icx285c.htm } http://www.atik-instruments.com/ } } Good luck, } } Austin } } P.S. I'm not financially involved with Artemis; but I do have one of } their excellent cameras. } } } Organization: NDT } } } } Title-Subject: [Filtered] price of used ccd camera } } } } Question: Hi, } } } } we are looking for a pre-owned ccd camera, such as optronics, } } Qimaging, Cooke, Roper, etc that can support our brightfield and/or } } fluorescent digital imagaing and quantitative analysis (volumentric } } analysis and cell, spine counting). Due to our budget limit, we can } } only offer less than USD1500. Please let me know if you have } } available one for sale or know the right party we can contact with, } } thanks! } } } } Mike } } } } -------------------------------------------------------------------- } } ------- } }
-- 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 Nov 28 14:04:05 2006 10, 27 -- Received: from harpo.itss.auckland.ac.nz (mailhost-old.auckland.ac.nz [130.216.190.13]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kASK44C9024006 10, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 14:04:04 -0600 10, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 10, 27 -- by harpo.itss.auckland.ac.nz (Postfix) with ESMTP id 5BB0F34E30; 10, 27 -- Wed, 29 Nov 2006 09:04:03 +1300 (NZDT) 10, 27 -- Received: from harpo.itss.auckland.ac.nz ([127.0.0.1]) 10, 27 -- by localhost (smtpc.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 10, 27 -- with ESMTP id 22013-22; Wed, 29 Nov 2006 09:04:03 +1300 (NZDT) 10, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 10, 27 -- by harpo.itss.auckland.ac.nz (Postfix) with ESMTP id 29B9B34C07; 10, 27 -- Wed, 29 Nov 2006 09:04:03 +1300 (NZDT) 10, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 10, 27 -- To: auntdaisy-at-gmail.com 10, 27 -- Date: Wed, 29 Nov 2006 09:09:03 +1300 10, 27 -- MIME-Version: 1.0 10, 27 -- Subject: Re: [Microscopy] Re: viaWWW: used ccd camera 10, 27 -- Cc: microscopy-at-microscopy.com 10, 27 -- Message-ID: {456D4E2F.20779.D3D19-at-localhost} 10, 27 -- Priority: normal 10, 27 -- In-reply-to: {200611271425.kAREPLLD003633-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==============================
Two post-doctoral positions are open in the laboratory of Prof. H. Kumar Wickramasinghe at the University of California, Irvine. The field of research is the Development of Novel Scanning Probe Instruments in Biology. We are seeking skilled individuals with a solid background in Scanning Probe Microscopy, with specific focus on instrument development.
One position requires a Ph.D. in Electrical Engineering, Applied Physics or equivalent with an emphasis in Biology. http://www.eng.uci.edu/node/1103
One position requires a Ph.D. in Biophysics, Bioengineering, Biology or equivalent. http://www.eng.uci.edu/node/1105
Both positions require previous demonstrated accomplishments in scanning probe instrument development.
Application deadline: Open until filled
Please send your curriculum vitae, a list of publications, and names of at least three references to:
Professor H. Kumar Wickramasinghe Department of Electrical Engineering and Computer Science 325 Engineering Tower University of California, Irvine Irvine, CA 92696-2175
Alternatively, materials may be submitted electronically to: Professor H. Kumar Wickramasinghe at: hkwick-at-uci.edu
The University of California, Irvine is an equal opportunity employer committed to excellence through diversity.
==============================Original Headers============================== 13, 21 -- From kerem.unal-at-sbcglobal.net Tue Nov 28 15:57:32 2006 13, 21 -- Received: from smtp109.sbc.mail.mud.yahoo.com (smtp109.sbc.mail.mud.yahoo.com [68.142.198.208]) 13, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kASLvTLe005069 13, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 15:57:31 -0600 13, 21 -- Received: (qmail 83042 invoked from network); 28 Nov 2006 21:57:26 -0000 13, 21 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 13, 21 -- s=s1024; d=sbcglobal.net; 13, 21 -- h=Received:X-YMail-OSG:Mime-Version:Content-Transfer-Encoding:Message-Id:Content-Type:To:From:Subject:Date:X-Mailer; 13, 21 -- b=Cxndd+lNLpEhKFihvzOqkpbfNH2eMMlysku+X8Y5xxyoskAaVhls//w44kNGds9zqzo5737XTlwYiVnLQBfMbXEepTCRfMcgSHWtEFvYQLZfQYhlL/Omn67qHI3wOQaHy3xxbNyjQKDlEWUfBAQRbZZHwgllte3nxxAjlYo1Jkc= ; 13, 21 -- Received: from unknown (HELO ?128.200.211.163?) (kerem.unal-at-sbcglobal.net-at-128.200.211.163 with plain) 13, 21 -- by smtp109.sbc.mail.mud.yahoo.com with SMTP; 28 Nov 2006 21:57:26 -0000 13, 21 -- X-YMail-OSG: vkNzBH8VM1lmdspvE9aFCGVhYB7gYgVNBhHf_.o04Rgei1laIVH_sQDutSgOC8E8nWa72NCMraLCSB948Nt44YAwvu2AHGv.9wgWX7cV5sAke5TQjPlsjQ-- 13, 21 -- Mime-Version: 1.0 (Apple Message framework v752.3) 13, 21 -- Content-Transfer-Encoding: 7bit 13, 21 -- Message-Id: {B6027B1E-E269-4709-8588-9E67F496A7F7-at-sbcglobal.net} 13, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 13, 21 -- To: Microscopy-at-microscopy.com 13, 21 -- From: Kerem Uenal {kerem.unal-at-sbcglobal.net} 13, 21 -- Subject: Two postdoctoral scholar positions available 13, 21 -- Date: Tue, 28 Nov 2006 13:57:15 -0800 13, 21 -- X-Mailer: Apple Mail (2.752.3) ==============================End of - Headers==============================
This message only applies to New Zealand laboratories.
Hello fellow New Zealand histologists
We urgently require some Shandon xylene substitute. Our order for new stock has been delayed and we have a researcher who is desperately seeking Shandon (sad title for a new movie maybe?). Can you help PLEASE!?!??
Kind Regards Matthew Downes -- Matthew Downes Electron Microscopy Technician Department of Zoology University of Otago P.O.Box 56 Dunedin New Zealand.
It doesn't say (or I couldn't find) what formats for images are supported. If it is JPG, you have compression artifacts in addition to the interpolation. If you do put that on a microscope, let me know how it works.
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: r.sims-at-auckland.ac.nz [mailto:r.sims-at-auckland.ac.nz] Sent: Tuesday, November 28, 2006 13:09 To: Mike Bode
Has anyone yet tried the newish Microsoft VX-6000 Webcam? It claims to give 5MP for stills, it's cheap, I'm tempted to get one, rip the lens off, and patch it onto the top of a microscope with sticky tape. Not much to lose.
cheers
rtch
O} } Dear Mike, } } Do you want colour or B&W + filters? } } Have you thought of an Astronomical camera? $1500 is tight, but } it } might cover an Artemis camera. I'd recommend one of the better Sony } chips, e.g. ICX285AL or ICX285AQ. } http://www.artemisccd.co.uk/icx285c.htm } http://www.atik-instruments.com/ } } Good luck, } } Austin } } P.S. I'm not financially involved with Artemis; but I do have one of } their excellent cameras. } } } Organization: NDT } } } } Title-Subject: [Filtered] price of used ccd camera } } } } Question: Hi, } } } } we are looking for a pre-owned ccd camera, such as optronics, } } Qimaging, Cooke, Roper, etc that can support our brightfield and/or } } fluorescent digital imagaing and quantitative analysis (volumentric } } analysis and cell, spine counting). Due to our budget limit, we can
} } only offer less than USD1500. Please let me know if you have } } available one for sale or know the right party we can contact with, } } thanks! } } } } Mike } } } } -------------------------------------------------------------------- } } ------- } }
-- 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 Nov 28 14:04:05 2006 10, 27 -- Received: from harpo.itss.auckland.ac.nz (mailhost-old.auckland.ac.nz [130.216.190.13]) 10, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kASK44C9024006 10, 27 -- for {microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 14:04:04 -0600 10, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 10, 27 -- by harpo.itss.auckland.ac.nz (Postfix) with ESMTP id 5BB0F34E30; 10, 27 -- Wed, 29 Nov 2006 09:04:03 +1300 (NZDT) 10, 27 -- Received: from harpo.itss.auckland.ac.nz ([127.0.0.1]) 10, 27 -- by localhost (smtpc.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 10, 27 -- with ESMTP id 22013-22; Wed, 29 Nov 2006 09:04:03 +1300 (NZDT) 10, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 10, 27 -- by harpo.itss.auckland.ac.nz (Postfix) with ESMTP id 29B9B34C07; 10, 27 -- Wed, 29 Nov 2006 09:04:03 +1300 (NZDT) 10, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 10, 27 -- To: auntdaisy-at-gmail.com 10, 27 -- Date: Wed, 29 Nov 2006 09:09:03 +1300 10, 27 -- MIME-Version: 1.0 10, 27 -- Subject: Re: [Microscopy] Re: viaWWW: used ccd camera 10, 27 -- Cc: microscopy-at-microscopy.com 10, 27 -- Message-ID: {456D4E2F.20779.D3D19-at-localhost} 10, 27 -- Priority: normal 10, 27 -- In-reply-to: {200611271425.kAREPLLD003633-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==============================
==============================Original Headers============================== 25, 24 -- From Mike.Bode-at-olympus-sis.com Tue Nov 28 17:23:41 2006 25, 24 -- Received: from mail.soft-imaging.de (ns.soft-imaging.de [62.180.61.130]) 25, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kASNNeiZ028611 25, 24 -- for {Microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 17:23:41 -0600 25, 24 -- Received: from muenster.soft-imaging.net (muenster.soft-imaging.de [10.26.20.9]) 25, 24 -- by mail.soft-imaging.de (8.13.3/8.13.3/SuSE Linux 0.7) with ESMTP id kASNO7O1019517 25, 24 -- for {Microscopy-at-microscopy.com} ; Wed, 29 Nov 2006 00:24:09 +0100 25, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 25, 24 -- Content-class: urn:content-classes:message 25, 24 -- MIME-Version: 1.0 25, 24 -- Content-Type: text/plain; 25, 24 -- charset="us-ascii" 25, 24 -- Subject: RE: [Microscopy] viaWWW: used ccd camera 25, 24 -- Date: Wed, 29 Nov 2006 00:17:06 +0100 25, 24 -- Message-ID: {78B53BA1C5A2D9449EDA30A98800EC9462733C-at-ms-s-gws.soft-imaging.net} 25, 24 -- In-Reply-To: {200611282008.kASK8pmj032585-at-ns.microscopy.com} 25, 24 -- X-MS-Has-Attach: 25, 24 -- X-MS-TNEF-Correlator: 25, 24 -- Thread-Topic: [Microscopy] viaWWW: used ccd camera 25, 24 -- Thread-Index: AccTKQhK9v1TqI15Qr6b3migYulMYQAGVveg 25, 24 -- From: "Mike Bode" {Mike.Bode-at-olympus-sis.com} 25, 24 -- To: {Microscopy-at-microscopy.com} 25, 24 -- Content-Transfer-Encoding: 8bit 25, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kASNNeiZ028611 ==============================End of - Headers==============================
for $1,500 you might take a look at the Nikon D200 - Street pricing seems to be } $1400 but then you'd also need the opticla coupling if you no not already a 35mm / f-mount setup. The D200 has a very low vibration mirror/shutter system and mirror-up shutter delay on it (10.2megapixel ). Our tests here shows it does well on even light weight scopes (Nikon optiphot), and great job with general fluorescence. However it does not integrate with quantitative 3-D rendering software packages.
On 27 Nov 2006 at 3:32, neurowu-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 } ---------------------------------------------------------------------------- } } 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 neurowu-at-yahoo.com as well as the MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: neurowu-at-yahoo.com } Name: mike } } Organization: NDT } } Title-Subject: [Filtered] price of used ccd camera } } Question: Hi, } } we are looking for a pre-owned ccd camera, such as optronics, } Qimaging, Cooke, Roper, etc that can support our brightfield and/or } fluorescent digital imagaing and quantitative analysis (volumentric } analysis and cell, spine counting). Due to our budget limit, we can } only offer less than USD1500. Please let me know if you have } available one for sale or know the right party we can contact with, } thanks! } } Mike } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 8, 12 -- From zaluzec-at-microscopy.com Mon Nov 27 03:31:04 2006 } 8, 12 -- Received: from [10.241.76.246] (mk194048137133.a1.net [194.48.137.133]) } 8, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAR9V3G0016973 } 8, 12 -- for {microscopy-at-microscopy.com} ; Mon, 27 Nov 2006 03:31:03 -0600 } 8, 12 -- Mime-Version: 1.0 } 8, 12 -- X-Sender: (Unverified) } 8, 12 -- Message-Id: {p06020402c19060606e5d-at-[10.241.76.246]} } 8, 12 -- Date: Mon, 27 Nov 2006 03:29:36 -0600 } 8, 12 -- To: microscopy-at-microscopy.com } 8, 12 -- From: neurowu-at-yahoo.com (by way of MicroscopyListserver) } 8, 12 -- Subject: viaWWW: used ccd camera } 8, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers==============================
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 9, 32 -- From edelmare-at-muohio.edu Wed Nov 29 14:26:44 2006 9, 32 -- Received: from spamfirewall.muohio.edu (wallaby.mcs.muohio.edu [134.53.6.28]) 9, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kATKQh0l010967 9, 32 -- for {microscopy-at-Microscopy.com} ; Wed, 29 Nov 2006 14:26:44 -0600 9, 32 -- X-ASG-Debug-ID: 1164832003-436c004a0000-Dem1zR 9, 32 -- X-Barracuda-URL: http://spamfirewall.muohio.edu:80/cgi-bin/mark.cgi 9, 32 -- X-Barracuda-Connect: mulnx23.mcs.muohio.edu[134.53.6.10] 9, 32 -- X-Barracuda-Start-Time: 1164832003 9, 32 -- Received: from mulnx23.mcs.muohio.edu (mulnx23.mcs.muohio.edu [134.53.6.10]) 9, 32 -- by spamfirewall.muohio.edu (Spam Firewall) with ESMTP 9, 32 -- id 40A23A0142; Wed, 29 Nov 2006 15:26:43 -0500 (EST) 9, 32 -- Received: from [192.168.1.23] ([134.53.14.105]) 9, 32 -- by mulnx23.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id kATKQh2S014041; 9, 32 -- Wed, 29 Nov 2006 15:26:43 -0500 9, 32 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 9, 32 -- To: neurowu-at-yahoo.com 9, 32 -- Date: Wed, 29 Nov 2006 15:26:42 -0500 9, 32 -- MIME-Version: 1.0 9, 32 -- X-ASG-Orig-Subj: Re: [Microscopy] viaWWW: used ccd camera 9, 32 -- Subject: Re: [Microscopy] viaWWW: used ccd camera 9, 32 -- CC: microscopy-at-Microscopy.com 9, 32 -- Message-ID: {456DA6B2.30327.17AE951-at-edelmare.muohio.edu} 9, 32 -- Priority: normal 9, 32 -- In-reply-to: {200611270932.kAR9WH2H018275-at-ns.microscopy.com} 9, 32 -- References: {200611270932.kAR9WH2H018275-at-ns.microscopy.com} 9, 32 -- X-mailer: Pegasus Mail for Windows (4.41) 9, 32 -- Content-type: text/plain; charset=US-ASCII 9, 32 -- Content-transfer-encoding: 7BIT 9, 32 -- Content-description: Mail message body 9, 32 -- X-Barracuda-Virus-Scanned: by Barracuda Spam Firewall at muohio.edu 9, 32 -- X-Barracuda-Spam-Score: 0.00 9, 32 -- X-Barracuda-Spam-Status: No, SCORE=0.00 using per-user scores of TAG_LEVEL=3.5 QUARANTINE_LEVEL=1000.0 KILL_LEVEL=1000.0 ==============================End of - Headers==============================
Greetings Everyone, Recently, one of the three 24 volt lens power supplies on our five year old TF20 died. We found out that 24 volt replacements are no longer available and have not been manufactured for 5 or 6 years. The current replacement is two 38 volt power supplies. Consequently, we have two spare 24 volt units that we will not need. If someone out there would like to have them, please contact me. We just ask that you pay the shipping. Don
Mr. Donald Gantz Research Histologist/Electron Microscopist Dept. Physiology and Biophysics Center for Advanced Biomedical Research Boston University School of Medicine 715 Albany Street Boston, MA 02118 phone: 617-638-4017
==============================Original Headers============================== 3, 24 -- From gantz-at-bu.edu Wed Nov 29 14:39:38 2006 3, 24 -- Received: from relay9.bu.edu (relay9.bu.edu [128.197.27.239]) 3, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kATKdcGA022007 3, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Nov 2006 14:39:38 -0600 3, 24 -- X-Envelope-From: gantz-at-bu.edu 3, 24 -- Received: from biophysics1.bumc.bu.edu (biophysics1.bumc.bu.edu [155.41.209.120]) 3, 24 -- by relay9.bu.edu (8.13.8/8.13.8) with ESMTP id kATKdIs4022494 3, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Nov 2006 15:39:18 -0500 3, 24 -- Received: from gantz (dhcp209-127.bumc.bu.edu [155.41.209.127]) 3, 24 -- by biophysics1.bumc.bu.edu (8.13.8/8.12.11) with SMTP id kATKdIFw003810 3, 24 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 29 Nov 2006 15:39:18 -0500 3, 24 -- Message-ID: {005e01c713f6$c4ff3290$7fd1299b-at-gantz} 3, 24 -- From: "Don Gantz" {gantz-at-bu.edu} 3, 24 -- To: {Microscopy-at-MSA.Microscopy.Com} 3, 24 -- Subject: TF20 24volt power supplies 3, 24 -- Date: Wed, 29 Nov 2006 15:41:38 -0500 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 3, 24 -- X-MSMail-Priority: Normal 3, 24 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1807 3, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 ==============================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 doodyl-at-west-greene.k12.pa.us as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: West Greene High School in SW PA has a pSEM (EDS) operated by and for the students. It was purchased through a grant from the Appalachian Regional Commission and the help and continuing support of the RJ Lee Group, Inc. I am the Chemistry teacher and Curriculum Consultant responsible for planning and implementing curriculum with the SEM. Our current Grant through the ARC is to train other teachers in the tri-county area in using the SEM in the classroom; virtually, remotely, or physically through a Workshop in March-07, offered to the tri-county teachers (Greene, Washington, Fayette). Because this program is relatively new, I am open to ideas, coments, etc. Please feel free to contact me at any time. 724-499-5782 at the West Greene SEM Lab. Lurea Doody
"really old) information"??? Hey now I love the MT-2's! (The MT-2b is my favorite). Still have one in my lab. Did you get an answer to your question? If not I can work one up for you.
But basically at 500nm (0.5um) if you have to be accurate of 500nm - you need to use the section color. The adjustment wheel (On top of microtome) works interactively with the Thickness wheel on left front side (and they are calibrated in angstroms = 0.5um = 500nm = 5000Å). HOWEVER, the zero point of the calibration of the top knob is usually very far off if anyone has ever taken the case cover off the microtome.
On 21 Nov 2006 at 8:30, 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 } ---------------------------------------------------------------------------- } } } X-from: "Dr. Michael Horst" {HORST_MN-at-Mercer.edu} } To: "Greg Erdos" {gwe-at-ufl.edu} } Sent: Tuesday, November 21, 2006 9:00 AM } Subject: Help with an MT-2 ultramicrotome } } } } Hi- } } I would like to ask if anyone can help me understand the relationship } } between the adjustment knob and the five horizontal lines scribed into the } } shaft of the pivot arm. My goal is to cut 0.5 micron sections but the } } Instruction Manual does not address the use of these five slashes and how } } they relate to the adustment knob. } } Any one out there remember this (really old) information ? If so, please } } email me at horst_mn-at-mercer.edu } } Thanks! } } } } Mike } } } } Michael N. Horst } } School of Medicine } } Mercer University } } 1550 College St. } } Macon GA 31207 } } } } } } } ==============================Original Headers============================== } 5, 27 -- From gwe-at-ufl.edu Tue Nov 21 08:30:13 2006 } 5, 27 -- Received: from imf20aec.mail.bellsouth.net (imf20aec.mail.bellsouth.net [205.152.59.68]) } 5, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kALEUDli009598 } 5, 27 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 08:30:13 -0600 } 5, 27 -- Received: from ibm64aec.bellsouth.net ([70.152.55.172]) } 5, 27 -- by imf20aec.mail.bellsouth.net with ESMTP } 5, 27 -- id {20061121143012.QISX23001.imf20aec.mail.bellsouth.net-at-ibm64aec.bellsouth.net} } 5, 27 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 09:30:12 -0500 } 5, 27 -- Received: from ufcdd0b59c8a4d ([70.152.55.172]) by ibm64aec.bellsouth.net } 5, 27 -- with SMTP } 5, 27 -- id {20061121143012.QLXU16860.ibm64aec.bellsouth.net-at-ufcdd0b59c8a4d} } 5, 27 -- for {microscopy-at-microscopy.com} ; Tue, 21 Nov 2006 09:30:12 -0500 } 5, 27 -- Message-ID: {00b301c70d79$87f0af10$4d399846-at-ufcdd0b59c8a4d} } 5, 27 -- From: "greg erdos" {gwe-at-ufl.edu} } 5, 27 -- To: {microscopy-at-microscopy.com} } 5, 27 -- Subject: Fw: Help with an MT-2 ultramicrotome } 5, 27 -- Date: Tue, 21 Nov 2006 09:30:02 -0500 } 5, 27 -- MIME-Version: 1.0 } 5, 27 -- Content-Type: text/plain; } 5, 27 -- format=flowed; } 5, 27 -- charset="iso-8859-1"; } 5, 27 -- reply-type=original } 5, 27 -- Content-Transfer-Encoding: 7bit } 5, 27 -- X-Priority: 3 } 5, 27 -- X-MSMail-Priority: Normal } 5, 27 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2905 } 5, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 } ==============================End of - Headers==============================
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Director 364 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
"A main-frame: The biggest PC peripheral available."
==============================Original Headers============================== 11, 33 -- From edelmare-at-muohio.edu Thu Nov 30 10:11:16 2006 11, 33 -- Received: from spamfirewall.muohio.edu (wallaby.mcs.muohio.edu [134.53.6.28]) 11, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAUGBG2s006889 11, 33 -- for {microscopy-at-Microscopy.com} ; Thu, 30 Nov 2006 10:11:16 -0600 11, 33 -- X-ASG-Debug-ID: 1164903075-209b01090000-Dem1zR 11, 33 -- X-Barracuda-URL: http://spamfirewall.muohio.edu:80/cgi-bin/mark.cgi 11, 33 -- X-Barracuda-Connect: mulnx24.mcs.muohio.edu[134.53.6.11] 11, 33 -- X-Barracuda-Start-Time: 1164903075 11, 33 -- Received: from mulnx24.mcs.muohio.edu (mulnx24.mcs.muohio.edu [134.53.6.11]) 11, 33 -- by spamfirewall.muohio.edu (Spam Firewall) with ESMTP 11, 33 -- id E5097B7D25; Thu, 30 Nov 2006 11:11:15 -0500 (EST) 11, 33 -- Received: from [192.168.1.23] ([134.53.14.105]) 11, 33 -- by mulnx24.mcs.muohio.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id kAUGBFDS004845; 11, 33 -- Thu, 30 Nov 2006 11:11:15 -0500 11, 33 -- From: "Richard E. Edelmann" {edelmare-at-muohio.edu} 11, 33 -- To: HORST_MN-at-Mercer.edu 11, 33 -- Date: Thu, 30 Nov 2006 11:11:14 -0500 11, 33 -- MIME-Version: 1.0 11, 33 -- X-ASG-Orig-Subj: Re: [Microscopy] Fw: Help with an MT-2 ultramicrotome 11, 33 -- Subject: Re: [Microscopy] Fw: Help with an MT-2 ultramicrotome 11, 33 -- CC: gwe-at-ufl.edu, microscopy-at-Microscopy.com 11, 33 -- Message-ID: {456EBC52.17818.5B75BF6-at-edelmare.muohio.edu} 11, 33 -- Priority: normal 11, 33 -- In-reply-to: {200611211430.kALEUpgj010883-at-ns.microscopy.com} 11, 33 -- References: {200611211430.kALEUpgj010883-at-ns.microscopy.com} 11, 33 -- X-mailer: Pegasus Mail for Windows (4.41) 11, 33 -- Content-type: text/plain; charset=ISO-8859-1 11, 33 -- Content-description: Mail message body 11, 33 -- X-Barracuda-Virus-Scanned: by Barracuda Spam Firewall at muohio.edu 11, 33 -- X-Barracuda-Spam-Score: 0.00 11, 33 -- X-Barracuda-Spam-Status: No, SCORE=0.00 using per-user scores of TAG_LEVEL=3.5 QUARANTINE_LEVEL=1000.0 KILL_LEVEL=1000.0 11, 33 -- Content-Transfer-Encoding: 8bit 11, 33 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id kAUGBG2s006889 ==============================End of - Headers==============================
We have a field emission SEM that has massive contamination which appears as a yellowish residue in the specimen chamber and all the way up the column to the gun.
Mass spectroscopy of the residue gave the following results:
We have a number of users looking at polymers, the most likely suspects among the samples that go into the microscope, but so far there do not seem to be any that would result in the above contamination.
The real mystery is 1) all these samples are sputter coated with at least 3nm Pt before imaging. Considering the mobility of the contamination you would think problems would show up with the sputter coater even though vacuum and plasma energy are less than what the samples experience in the FESEM (most people are using low accelerating voltages of 1-5 keV by the way) 2)The yellowish color of the contamination seems odd, most of the samples are grey or black 3)The evidence at this point indicates a rapid rather than gradual rate of contamination, considering the amount of contamination it would seem an entire sample would have to vaporize before the user's eyes to provide enough material (of course they would have to admit to seeing this)
Has anyone else experienced anything like this (the rather seasoned service engineer has not)?
Many thanks for taking the time to read all of this.
Tom
} Tom Stephens } Assistant Research Scientist } Microscopy and Imaging Center } Texas A&M University } College Station, TX 77843 } office phone:979-845-1165 } email:tstephen-at-mic.tamu.edu
==============================Original Headers============================== 11, 20 -- From tstephen-at-mic.tamu.edu Thu Nov 30 11:48:14 2006 11, 20 -- Received: from sr-5-int.cis.tamu.edu (smtp-relay.tamu.edu [165.91.22.120]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAUHmEQ6019711 11, 20 -- for {Microscopy-at-Microscopy.Com} ; Thu, 30 Nov 2006 11:48:14 -0600 11, 20 -- Received: from localhost (localhost.tamu.edu [127.0.0.1]) 11, 20 -- by sr-5-int.cis.tamu.edu (Postfix) with ESMTP id 39F6C6AEF 11, 20 -- for {Microscopy-at-Microscopy.Com} ; Thu, 30 Nov 2006 11:48:14 -0600 (CST) 11, 20 -- Received: from jntcspc.mic.tamu.edu (cyn6400.tamu.edu [165.91.109.143]) 11, 20 -- by sr-5-int.cis.tamu.edu (Postfix) with ESMTP id AEC37730A 11, 20 -- for {Microscopy-at-Microscopy.Com} ; Thu, 30 Nov 2006 11:48:11 -0600 (CST) 11, 20 -- Message-Id: {5.2.0.9.0.20061129143701.02b26ef8-at-mic.tamu.edu} 11, 20 -- X-Sender: tstephen-at-mic.tamu.edu (Unverified) 11, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.0.9 11, 20 -- Date: Thu, 30 Nov 2006 11:48:13 -0600 11, 20 -- To: Microscopy-at-Microscopy.Com 11, 20 -- From: Tom Stephens {tstephen-at-mic.tamu.edu} 11, 20 -- Subject: SEM - Mystery contamination of FESEM 11, 20 -- Mime-Version: 1.0 11, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 11, 20 -- X-Virus-Scanned: amavisd-new at tamu.edu ==============================End of - Headers==============================
} } Email: doodyl-at-west-greene.k12.pa.us } Name: Lurea Doody } } Organization: West Greene HS, Waynesburg College } } Title-Subject: [Filtered] SEM in my High School } } Question: West Greene High School in SW PA has a pSEM (EDS) operated } by and for the students. It was purchased through a grant from the } Appalachian Regional Commission and the help and continuing support } of the RJ Lee Group, Inc. I am the Chemistry teacher and Curriculum } Consultant responsible for planning and implementing curriculum with } the SEM. Our current Grant through the ARC is to train other teachers } in the tri-county area in using the SEM in the classroom; virtually, } remotely, or physically through a Workshop in March-07, offered to } the tri-county teachers (Greene, Washington, Fayette). Because this } program is relatively new, I am open to ideas, coments, etc. Please } feel free to contact me at any time. 724-499-5782 at the West Greene } SEM Lab. Lurea Doody } } --------------------------------------------------------------------------- } Laura -
Project MICRO is for middle schools, but I try to track what's happening at the high school level. Here's another SEM in a Pennsylvania high school:
"The 2 teachers who have been primarily in charge are Heather Fogel (hef-at-rlasd.k12.pa.us) and Marcie Walizer (mlw-at-rlasd.k12.pa.us). They both teach biology at Red Lion Area Sr. High School." ___________________ And this is from a contact in Ohio:
"To answer your questions, 1) Yes, the van is still active - it's called Tech Trek [A SEM in a truck] 2) There is an SEM still located at Patterson Co-Op High School but not sure if anyone still uses it. The base [Wright-Patterson AFB] also still runs it's SEMeDS program where once a month, classes are brought in the afternoons from 3-6PM to do SEM.
The contact for Tech Trek is: Sharon Nelson (937) 904-8626 Sharon.Nelson-at-wpafb.af.mil The contact for SEMeDS is: Kim Stultz (937) 674-8623 Kimberly.Stultz-at-wpafb.af.mil " _____________________
"David L. Jones" {dljones-at-bestweb.net} is trying to start a SEM program in New York state.
If you get any other responses to your inquiry, will you please let me know? A comprehensive list would help everyone.
-- 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============================== 8, 18 -- From schooley-at-mcn.org Thu Nov 30 12:46:00 2006 8, 18 -- Received: from dns4.mcn.org (dns4.mcn.org [216.150.240.31]) 8, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAUIk05T031504 8, 18 -- for {microscopy-at-microscopy.com} ; Thu, 30 Nov 2006 12:46:00 -0600 8, 18 -- Received: from [66.52.139.70] (helo=[10.0.1.2]) 8, 18 -- by dns4.mcn.org with esmtpa (Exim 4.60) 8, 18 -- (envelope-from {schooley-at-mcn.org} ) 8, 18 -- id J9K44M-0003M3-3D; Thu, 30 Nov 2006 10:45:59 -0800 8, 18 -- Mime-Version: 1.0 8, 18 -- Message-Id: {a06200704c194cc3c076a-at-[10.0.1.2]} 8, 18 -- In-Reply-To: {200611300845.kAU8jUV2024078-at-ns.microscopy.com} 8, 18 -- References: {200611300845.kAU8jUV2024078-at-ns.microscopy.com} 8, 18 -- Date: Thu, 30 Nov 2006 10:27:06 -0800 8, 18 -- To: doodyl-at-west-greene.k12.pa.us 8, 18 -- From: Caroline Schooley {schooley-at-mcn.org} 8, 18 -- Subject: Re: [Microscopy] viaWWW: SEM in my High School 8, 18 -- Cc: microscopy-at-microscopy.com, tpepper-at-iastate.edu 8, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
And I'm sure there must be numerous other commercial sources. (By the way, I have no financial interest in either firm, but have made satisfactory small purchases from both.)
-------Roy Arrowood
==============================Original Headers============================== 7, 20 -- From arrowood-at-utep.edu Thu Nov 30 15:31:47 2006 7, 20 -- Received: from itdsrvmail00.utep.edu (itdsrvmail00.utep.edu [129.108.0.100]) 7, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kAULVl4C013220 7, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 30 Nov 2006 15:31:47 -0600 7, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 20 -- Content-class: urn:content-classes:message 7, 20 -- MIME-Version: 1.0 7, 20 -- Content-Type: text/plain; 7, 20 -- charset="us-ascii" 7, 20 -- Subject: in reply to "source for OM sample slides" 7, 20 -- Date: Thu, 30 Nov 2006 14:31:46 -0700 7, 20 -- Message-ID: {8B48F5AF730998488AF7BC9D7D22172001D50946-at-itdsrvmail00.utep.edu} 7, 20 -- X-MS-Has-Attach: 7, 20 -- X-MS-TNEF-Correlator: 7, 20 -- Thread-Topic: in reply to "source for OM sample slides" 7, 20 -- Thread-Index: AccUxvAlZ80lyzGWTEe6c0IXXnyV+g== 7, 20 -- From: "Arrowood, Roy" {arrowood-at-utep.edu} 7, 20 -- To: {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 kAULVl4C013220 ==============================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 yxu-at-thinfilm.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: yxu-at-thinfilm.com Name: yang xu
Organization: Applied ThinFilm Products
Title-Subject: [Filtered] Help to obtain Manuals for Cambridge S-600 SEM
Question: Please help us to obtain the User Manual and the Maintenance Manual for Cambridge S-600 SEM. Your kind help are greatly appreciated.
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 phankus-at-yahoo.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: phankus-at-yahoo.com Name: pankaj
Organization: UGC INDORE INDIA
Title-Subject: [Filtered] ELECTRON DIFFRACTION SIMULATION SOFTWARE
Question: I AM LOOKING FOR A FREE ELECTRON DIFFRACTION SIMULATION PROGRAM/SOFTWARE (WINDOWS BASED) PLEASE LET ME KNOW FROM WHERE TO GET THE SAME. WITH REGARDS PANKAJ
On Dec 1, 2006, at 3:52 AM, phankus-at-yahoo.com wrote:
} Organization: UGC INDORE INDIA } } Title-Subject: [Filtered] ELECTRON DIFFRACTION SIMULATION SOFTWARE } } Question: I AM LOOKING FOR A FREE ELECTRON DIFFRACTION SIMULATION } PROGRAM/SOFTWARE (WINDOWS BASED) } PLEASE LET ME KNOW FROM WHERE TO GET THE SAME. } WITH REGARDS } PANKAJ } Dear Pankaj, The following was posted on the list on 25 Feb, 2003:
} I favor the plane-wave multislice implementation by Earl Kirkland. In } the frozen phonon approximation, it is arguably the most accurate } algorithm (see D. A. Muller et al Ultramicroscopy 86, 371 (2001)). } Best of all, you get the entire source code and executables for Mac } and Windows for the price of Kirkland's book! The book is "Advanced } Computing in Electron Microscopy", by Earl J. Kirkland, Plenum 1998, } ISBN 0-306-45936-1. It contains a concise summary of electron } scattering and image formation, an extensive treatment of the } plane-wave multislice image simulation method, and advice for doing } accurate simulations with examples. } } This package has one drawback for some users: the user-interface is } command-line only. There is no slick GUI and no graphical help } constructing atomic models. } } } } Good luck! } 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 } It's not quite free, but the book may be worth it to you. 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, 22 -- From tivol-at-caltech.edu Fri Dec 1 12:15:17 2006 5, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 5, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB1IFHkZ011398 5, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 12:15:17 -0600 5, 22 -- Received: from fire-dog.caltech.edu (fire-dog [192.168.1.4]) 5, 22 -- by water-ox-postvirus (Postfix) with ESMTP 5, 22 -- id 83F3C2EFB5; Fri, 1 Dec 2006 10:15:12 -0800 (PST) 5, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 5, 22 -- by water-ox.its.caltech.edu (Postfix) with ESMTP 5, 22 -- id E928D2EF14; Fri, 1 Dec 2006 10:15:07 -0800 (PST) 5, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 5, 22 -- In-Reply-To: {200612011152.kB1BqvFN022849-at-ns.microscopy.com} 5, 22 -- References: {200612011152.kB1BqvFN022849-at-ns.microscopy.com} 5, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 5, 22 -- Message-Id: {945f7a0f92105c08c9f0518f97fdbbf6-at-caltech.edu} 5, 22 -- Content-Transfer-Encoding: 7bit 5, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 5, 22 -- Subject: Re: [Microscopy] viaWWW: ELECTRON DIFFRACTION SIMULATION SOFTWARE 5, 22 -- Date: Fri, 1 Dec 2006 10:21:09 -0800 5, 22 -- To: microscopy-at-msa.microscopy.com, phankus-at-yahoo.com 5, 22 -- X-Mailer: Apple Mail (2.624) 5, 22 -- X-Spam-Scanned: at Caltech-ITS on fire-dog by amavisd-2.3.3 ==============================End of - Headers==============================
We have a Gatan Model 655 Dry Pumping Station (www.gatan.com/holders/655_drypumpingstation.html) equipped with pods for storage of TEM specimens. In particular, TEM cross-sections of integrated circuits using copper interconnects seem to be particularly vulnerable to corrosion in atmosphere. This vacuum station has been essential in protecting the specimens that we put so much work into making, and has been very successful for quite some time. The dry vacuum system consists of a 'drag' pump, which is essentially a turbo pump, backed by a diaphragm type rough pump. Last summer, one of the rubber diaphragms failed in the backing pump, and the system ran for a few days with degraded vacuum. After the diaphragms were replaced and the high vacuum restored to the E-5 Torr range, we found that TEM cross-sections of integrated circuits using copper interconnects were destroyed in just a few hours if stored in this vacuum system. EDX of the copper corrosion product showed the presence of sulfur. We have tried cleaning the storage pods, and part of the inside of the vacuum chamber, but have been totally frustrated in being able to restore the essential function of this vacuum storage system. Any suggestions as to what may have happened, and what we can do to clean up the system would be very much appreciated. Thank you.
John Mardinly Intel Corporation
Disclaimer: This is a communication from this author and does not represent an opinion of Intel Corporation.
==============================Original Headers============================== 6, 30 -- From john.mardinly-at-intel.com Fri Dec 1 13:35:49 2006 6, 30 -- Received: from mga01.intel.com (mga01.intel.com [192.55.52.88]) 6, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB1JZnHb024448 6, 30 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 13:35:49 -0600 6, 30 -- Received: from fmsmga001.fm.intel.com ([10.253.24.23]) 6, 30 -- by mga01.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- Received: from scsmsx332.sc.intel.com (HELO scsmsx332.amr.corp.intel.com) ([10.3.90.6]) 6, 30 -- by fmsmga001.fm.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- X-ExtLoop1: 1 6, 30 -- X-IronPort-AV: i="4.09,486,1157353200"; 6, 30 -- d="scan'208"; a="171762165:sNHT19309206" 6, 30 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 30 -- Fri, 1 Dec 2006 11:35:31 -0800 6, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 30 -- Content-class: urn:content-classes:message 6, 30 -- MIME-Version: 1.0 6, 30 -- Content-Type: text/plain; 6, 30 -- charset="us-ascii" 6, 30 -- Subject: Contamination of Vacuum Specimen Storage System 6, 30 -- Date: Fri, 1 Dec 2006 11:35:31 -0800 6, 30 -- Message-ID: {1DF4C4D62339DB4C9C98DF04213995B601F0A84F-at-scsmsx413.amr.corp.intel.com} 6, 30 -- X-MS-Has-Attach: 6, 30 -- X-MS-TNEF-Correlator: 6, 30 -- Thread-Topic: Contamination of Vacuum Specimen Storage System 6, 30 -- Thread-Index: AccVcgLpZR8yy6qqRpi55yhxXPXijQAAIjDgAAAaArAAAVCSIAAB3rtQ 6, 30 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 6, 30 -- To: {Microscopy-at-msa.microscopy.com} 6, 30 -- X-OriginalArrivalTime: 01 Dec 2006 19:35:31.0926 (UTC) FILETIME=[DD2F8F60:01C7157F] 6, 30 -- Content-Transfer-Encoding: 8bit 6, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kB1JZnHb024448 ==============================End of - Headers==============================
I have a chemist who wants to look at nanoparticle distribution in fully hydrated acrylamide gel. Unfortunately the particles are too small to image using cryoFESEM. He wanted to just dump the stuff on a grid and put it in the TEM. After my emphatic "NO", I finally persuaded him that it would have to be dehydrated, embedded and thin sectioned. Of course there will be the inevitable collapse of the gel when dehydrated.
Does anyone have suggestions as to ways to fix the gel to try to stabilize it even a little bit prior to dehydration?
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 Fri Dec 1 13:44:05 2006 6, 21 -- Received: from 1061exfe04.adpc.purdue.edu (1061exfe04.adpc.purdue.edu [128.210.63.227]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB1Ji5Jr002829 6, 21 -- for {microscopy-at-microscopy.com} ; Fri, 1 Dec 2006 13:44:05 -0600 6, 21 -- Received: from exchange.purdue.edu ([172.21.6.23]) by 1061exfe04.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 6, 21 -- Fri, 1 Dec 2006 14:44:05 -0500 6, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 6, 21 -- Fri, 1 Dec 2006 19:44:05 +0000 6, 21 -- User-Agent: Microsoft-Entourage/11.2.5.060620 6, 21 -- Date: Fri, 01 Dec 2006 14:44:04 -0500 6, 21 -- Subject: Embedding acrylamide gel 6, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 6, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 6, 21 -- Message-ID: {C195F034.16EDA%dsherman-at-purdue.edu} 6, 21 -- Thread-Topic: Embedding acrylamide gel 6, 21 -- Thread-Index: AccVgQ5nTOjl94F0EduIegARJN08Mg== 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: 01 Dec 2006 19:44:05.0395 (UTC) FILETIME=[0F3CB630:01C71581] ==============================End of - Headers==============================
Is contamination dry or rather oily? Does it look like corrosion (exhibits any adhesion to the surfaces?) Does it prefer to precipitate on any particular materials or at any particular locations?
What vacuum pumps are in the system? Oil diffusion? Oil sealed rotary? Turbo? If fluids are present, what fluids?
Was anything done recently with water cooling system? Did cooling accidents happen? If ODP is present, is ODP cooling line connected in series with electronics cooling line, or in parallel?
If SEM has pneumatic valves, do you check condition of air supply regularly? If water condenses into pneumatic lines, valves will not necessarily fail at once, but they will slow down - that almost certainly will cause vac. fluids surges into high vacuum areas, while ODP fluid itself could be damaged to various degrees.
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: {tstephen-at-mic.tamu.edu} To: {vitalylazar-at-att.net} Sent: Thursday, November 30, 2006 12:50 PM
Laboratory Technician, Microscopy & Imaging Facility, American Museum Of Natural History.
The Laboratory Technician will be responsible for assisting and/or training scientific staff with Scanning Electron Microscopy, X-ray microanalysis, Laser Scanning Confocal Microscopy, Cathodoluminescence Spectroscopy, specimen preparation and image processing and printing; maintain functionality and capability of lab systems to suit the needs of the scientific staff and emerging technologies; interact with curatorial staff and students on diverse projects using microscopy and digital imaging in the fields of zoology, paleontology, anthropology, archeology, and earth and planetary sciences. The ideal candidate will possess knowledge of Scanning Electron Beam instruments and microanalysis and computer hardware/software; display patience, willingness to learn, flexibility, excellent organizational and intra-personal skills. At least one year of full-time Scanning Electron Microscopy experience; BS required; major in biology preferred.
Full time, 35 hrs/week, excellent benefits. Salary: $34,000/year. Please send resumé and cover letter along with 2 letters of recommendation to hrdesk-at-amnh.org All other inquiries should be sent to hrdesk-at-amnh.org The American Museum of Natural History is an Equal Opportunity Employer.
==============================Original Headers============================== 4, 24 -- From mey-at-amnh.org Fri Dec 1 16:06:20 2006 4, 24 -- Received: from lepore.amnh.org (lepore.amnh.org [216.73.241.12]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB1M6Kvj028039 4, 24 -- for {Microscopy-at-microscopy.org} ; Fri, 1 Dec 2006 16:06:20 -0600 4, 24 -- Received: from localhost (localhost [127.0.0.1]) 4, 24 -- by lepore.amnh.org (Postfix) with ESMTP id CBE6B5B768 4, 24 -- for {Microscopy-at-microscopy.org} ; Fri, 1 Dec 2006 17:06:19 -0500 (EST) 4, 24 -- X-Virus-Scanned: amavisd-new at amnh.org 4, 24 -- Received: from lepore.amnh.org ([127.0.0.1]) 4, 24 -- by localhost (lepore.amnh.org [127.0.0.1]) (amavisd-new, port 10024) 4, 24 -- with ESMTP id 631RpXF5I2vq for {Microscopy-at-microscopy.org} ; 4, 24 -- Fri, 1 Dec 2006 17:06:18 -0500 (EST) 4, 24 -- Received: from [172.16.60.30] (216-73-249-246.dynamic.amnh.org [216.73.249.246]) 4, 24 -- by lepore.amnh.org (Postfix) with ESMTP id CEFDA5B765 4, 24 -- for {Microscopy-at-microscopy.org} ; Fri, 1 Dec 2006 17:06:17 -0500 (EST) 4, 24 -- Message-ID: {4570A759.5030709-at-amnh.org} 4, 24 -- Date: Fri, 01 Dec 2006 17:06:17 -0500 4, 24 -- From: Jacob Louis Mey {mey-at-amnh.org} 4, 24 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) 4, 24 -- MIME-Version: 1.0 4, 24 -- To: Microscopy-at-microscopy.org 4, 24 -- Subject: Job Posting: Lab Technician, American Museum of Natural History 4, 24 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 24 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Including tannic acid in the primary fix, following with UrAc or OsO4, has a good reputation for enhancing resistance of lattices against shrinkage. We have used the TA-UrAc sequence in acetone for freeze-substitution of muscle fibers, slam frozen while supported on a thin underlying block of 2.5% agar gel. (Nylon spacers limited squashing during the impact). The agar gel mesh appeared quite reasonably preserved when sections were oriented to include it; I can't say we took any pictures of it, because our interest was in the muscle.
-mike reedy-
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************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
John, I think that the sulfur came from the rubber diaphragms. I'm surprised, because I thought that they would have been VitonR. Either that or lubrication oil from moving parts that came through the failed diaphragm and was exposed to the system. Was the diaphragm torn or had holes in it? Sulfur's atomic weight just might let it get through a turbo pump. Anyway, I think what you might be able to do is "getter" the sulfur in your system. Get some copper powder and put it in your system for a while. The smaller the powder particles and the more open the container, the better gettering that you will have. Silver ought to work also. It will take some time to getter the sulfur in your system out, but given some time, it should be able to do it. You will also be able to monitor the progress on the copper particles if you periodically change the copper.
In the mean time, you might want to consider storing your samples with our SampleSaverT system. We have a specially vented TEM storage box for the SampleSaverT container system that will take TEM grids. If the samples are FIB cuts, you can store them in the SampleSaverT container in our FortressT FIB holders that will also fit into the SampleSaverT container. The SampleSaverT containers worked for my cross section samples of Low-E glass where there are two layers of Ag exposed. Unfortunately, I haven't gotten any feedback yet from how well they work for copper metallization samples. I expect them to work as well there, but as I said, I don't have hard evidence, yet.
BTW, Generally, the suggested maintenance replacement time on diaphragms for pumps is about a year.
Disclaimer: South Bay Technology, Inc. makes and sells the SampleSaverT container and FortressT FIB holders.
-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: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, December 01, 2006 11:40 AM To: Walck-at-SouthBayTech.com
We have a Gatan Model 655 Dry Pumping Station (www.gatan.com/holders/655_drypumpingstation.html) equipped with pods for storage of TEM specimens. In particular, TEM cross-sections of integrated circuits using copper interconnects seem to be particularly vulnerable to corrosion in atmosphere. This vacuum station has been essential in protecting the specimens that we put so much work into making, and has been very successful for quite some time. The dry vacuum system consists of a 'drag' pump, which is essentially a turbo pump, backed by a diaphragm type rough pump. Last summer, one of the rubber diaphragms failed in the backing pump, and the system ran for a few days with degraded vacuum. After the diaphragms were replaced and the high vacuum restored to the E-5 Torr range, we found that TEM cross-sections of integrated circuits using copper interconnects were destroyed in just a few hours if stored in this vacuum system. EDX of the copper corrosion product showed the presence of sulfur. We have tried cleaning the storage pods, and part of the inside of the vacuum chamber, but have been totally frustrated in being able to restore the essential function of this vacuum storage system. Any suggestions as to what may have happened, and what we can do to clean up the system would be very much appreciated. Thank you.
John Mardinly Intel Corporation
Disclaimer: This is a communication from this author and does not represent an opinion of Intel Corporation.
==============================Original Headers============================== 6, 30 -- From john.mardinly-at-intel.com Fri Dec 1 13:35:49 2006 6, 30 -- Received: from mga01.intel.com (mga01.intel.com [192.55.52.88]) 6, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB1JZnHb024448 6, 30 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 13:35:49 -0600 6, 30 -- Received: from fmsmga001.fm.intel.com ([10.253.24.23]) 6, 30 -- by mga01.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- Received: from scsmsx332.sc.intel.com (HELO scsmsx332.amr.corp.intel.com) ([10.3.90.6]) 6, 30 -- by fmsmga001.fm.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- X-ExtLoop1: 1 6, 30 -- X-IronPort-AV: i="4.09,486,1157353200"; 6, 30 -- d="scan'208"; a="171762165:sNHT19309206" 6, 30 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 30 -- Fri, 1 Dec 2006 11:35:31 -0800 6, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 30 -- Content-class: urn:content-classes:message 6, 30 -- MIME-Version: 1.0 6, 30 -- Content-Type: text/plain; 6, 30 -- charset="us-ascii" 6, 30 -- Subject: Contamination of Vacuum Specimen Storage System 6, 30 --
Dear Dr. Mardinly, I would suspect that the rubber in the seal that failed is the source of the sulphur and this will react with the copper strongly. Unfortunately, this will be distributed throughout the system by the oil back-streamed by the system failure and can only be removed by very thorough cleaning of all chambers and lines. You might try baking the system after cleaning with solvents to get rig of all the oil. Change the oil in the backing pump and run everything for a week to clean it up. The other thing I have used successfully to clean up an SEM that was contaminating badly was to bleed in a stream of nitrogen gas through a feed through I made with a bit of fine stainless steel tubing and a metering valve. The gas valve was turned on until the SEM baffle valve closed and a bit more. Leave for about one hour. Take a blank port cover, drill a small hole through, glue the SS tubing through the hole, swage the fine metering valve on to the SS tubing. Attach the other side of the metering valve to a bag you can fill with nitrogen gas. A bag fill will last more than an hour at the right gas flow rate.) You can use a freshly-polished piece of copper to test your system. Good luck, Mary Mager Electron Microscopist Materials Eng. UBC #419 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA Phone: 604-822-5648 Fax: 604-822-3619 email: mager-at-interchange.ubc.ca
-----Original Message----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, December 01, 2006 11:42 AM To: mager-at-interchange.ubc.ca
We have a Gatan Model 655 Dry Pumping Station (www.gatan.com/holders/655_drypumpingstation.html) equipped with pods for storage of TEM specimens. In particular, TEM cross-sections of integrated circuits using copper interconnects seem to be particularly vulnerable to corrosion in atmosphere. This vacuum station has been essential in protecting the specimens that we put so much work into making, and has been very successful for quite some time. The dry vacuum system consists of a 'drag' pump, which is essentially a turbo pump, backed by a diaphragm type rough pump. Last summer, one of the rubber diaphragms failed in the backing pump, and the system ran for a few days with degraded vacuum. After the diaphragms were replaced and the high vacuum restored to the E-5 Torr range, we found that TEM cross-sections of integrated circuits using copper interconnects were destroyed in just a few hours if stored in this vacuum system. EDX of the copper corrosion product showed the presence of sulfur. We have tried cleaning the storage pods, and part of the inside of the vacuum chamber, but have been totally frustrated in being able to restore the essential function of this vacuum storage system. Any suggestions as to what may have happened, and what we can do to clean up the system would be very much appreciated. Thank you.
John Mardinly Intel Corporation
Disclaimer: This is a communication from this author and does not represent an opinion of Intel Corporation.
==============================Original Headers============================== 6, 30 -- From john.mardinly-at-intel.com Fri Dec 1 13:35:49 2006 6, 30 -- Received: from mga01.intel.com (mga01.intel.com [192.55.52.88]) 6, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB1JZnHb024448 6, 30 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 13:35:49 -0600 6, 30 -- Received: from fmsmga001.fm.intel.com ([10.253.24.23]) 6, 30 -- by mga01.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- Received: from scsmsx332.sc.intel.com (HELO scsmsx332.amr.corp.intel.com) ([10.3.90.6]) 6, 30 -- by fmsmga001.fm.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- X-ExtLoop1: 1 6, 30 -- X-IronPort-AV: i="4.09,486,1157353200"; 6, 30 -- d="scan'208"; a="171762165:sNHT19309206" 6, 30 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 30 -- Fri, 1 Dec 2006 11:35:31 -0800 6, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 30 -- Content-class: urn:content-classes:message 6, 30 -- MIME-Version: 1.0 6, 30 -- Content-Type: text/plain; 6, 30 -- charset="us-ascii" 6, 30 -- Subject: Contamination of Vacuum Specimen Storage System 6, 30 --
On Dec 1, 2006, at 11:44 AM, dsherman-at-purdue.edu wrote:
} Does anyone have suggestions as to ways to fix the gel to try to } stabilize } it even a little bit prior to dehydration? } Dear Debby, You could try high-pressure freezing followed by either freeze substitution or cryosectioning. For that matter, if the gel doesn't form ice crystals, just freezing it then cryosectioning or freeze-sub could work. If you could soak the gel in a cryoprotectant without disturbing the nanoparticles, then there should be no trouble with the freezing, but I don't know how the presence of the cryoprotectant would affect subsequent steps. 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 Dec 1 17:50:06 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 (8.12.11.20060308/8.12.8) with ESMTP id kB1No5GQ007704 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 17:50:05 -0600 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 4C59735818 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 15:50:05 -0800 (PST) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by wood-ox.its.caltech.edu (Postfix) with ESMTP id D588E2EEE2 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 15:50:03 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200612011944.kB1JiAeL002991-at-ns.microscopy.com} 4, 22 -- References: {200612011944.kB1JiAeL002991-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {778b3010dd77a689e15cffb02c3535d5-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Embedding acrylamide gel 4, 22 -- Date: Fri, 1 Dec 2006 15:56:05 -0800 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==============================
We had tried covering the nanowires with G1 epoxy and it works fine for us. After the glue cures, grind and flatten the surface, removing the excess glue until the glue is around 0.1mm or less thick. Then prepare the cross-sectional sample as it were a thin film sample. You can glue it to a piece of silicon wafer, slice and disk, then polish it with dimpling machine. During ion milling, you want to use sample oscillation, with glue line perpendicular to the beam. The ion beam should be only allowed to bombard the sample from the backside of the substrate.
Another method is FIB (Focused Ion Beam). We haven't used this method, but it should be practical if you have access to FIB.
Chengyu Song National Center For Electron Microscopy Lawrence Berkeley National Laboratory Berkeley, CA 94720
catherine.bougerol-at-cea.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 } ---------------------------------------------------------------------------- } } 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 catherine.bougerol-at-cea.fr as well as the } MIcroscopy Listserver } --------------------------------------------------------------------------- } } Email: catherine.bougerol-at-cea.fr } Name: catherine BOUGEROL } } Organization: CNRS } } Title-Subject: [Filtered] preparation of nanowires for TEM } } Question: I have got ZnSe nanowires grown on a GaAs substrate. Up to } now, I have prepared them by putting a piece of the sample in } methanol and letting it for one hour in an ultrasonic device ; them I } put a drop of the solution on a copper carbon grid. With this method, } the nanowires are broken and one cannot study the interface with the } substrate. This is why I am trying to find another method of } preparation keeping them on the substrate. } } --------------------------------------------------------------------------- } } ==============================Original Headers============================== } 6, 12 -- From zaluzec-at-microscopy.com Tue Nov 28 08:39:37 2006 } 6, 12 -- Received: from [128.130.45.29] (msdvpn8.msd.anl.gov [130.202.238.72]) } 6, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kASEdPan026530 } 6, 12 -- for {microscopy-at-microscopy.com} ; Tue, 28 Nov 2006 08:39:25 -0600 } 6, 12 -- Mime-Version: 1.0 } 6, 12 -- X-Sender: (Unverified) } 6, 12 -- Message-Id: {p06020406c191fa7b5b57-at-[128.130.45.29]} } 6, 12 -- Date: Tue, 28 Nov 2006 15:39:23 +0100 } 6, 12 -- To: microscopy-at-microscopy.com } 6, 12 -- From: catherine.bougerol-at-cea.fr (by way of MicroscopyListserver) } 6, 12 -- Subject: viaWWW: preparation of nanowires for TEM } 6, 12 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } ==============================End of - Headers==============================
==============================Original Headers============================== 7, 25 -- From csong-at-lbl.gov Fri Dec 1 18:40:43 2006 7, 25 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 7, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB20egSr019120 7, 25 -- for {microscopy-at-microscopy.com} ; Fri, 1 Dec 2006 18:40:42 -0600 7, 25 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 7, 25 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id kB20eeie012025 7, 25 -- for {microscopy-at-microscopy.com} ; Fri, 1 Dec 2006 16:40:41 -0800 (PST) 7, 25 -- Received: from lbl.gov (apple-0-5-2-e0-6a-f3.dhcp.lbl.gov [131.243.3.239]) 7, 25 -- by mta2.lbl.gov (8.13.8/8.13.8) with ESMTP id kB20edGN012021; 7, 25 -- Fri, 1 Dec 2006 16:40:39 -0800 (PST) 7, 25 -- Message-ID: {4570BD81.E85379F7-at-lbl.gov} 7, 25 -- Date: Fri, 01 Dec 2006 16:41:39 -0700 7, 25 -- From: chengyu song {csong-at-lbl.gov} 7, 25 -- Reply-To: csong-at-lbl.gov 7, 25 -- Organization: lbl 7, 25 -- X-Mailer: Mozilla 4.73C-CCK-MCD LBNL V4.73 Build 2 (Macintosh; U; PPC) 7, 25 -- X-Accept-Language: en,pdf 7, 25 -- MIME-Version: 1.0 7, 25 -- To: catherine.bougerol-at-cea.fr, microscopy-at-microscopy.com 7, 25 -- Subject: Re: [Microscopy] viaWWW: preparation of nanowires for TEM 7, 25 -- References: {200611281443.kASEhcEe032277-at-ns.microscopy.com} 7, 25 -- Content-Type: text/plain; charset=gb2312; x-mac-type="54455854"; x-mac-creator="4D4F5353" 7, 25 -- Content-Transfer-Encoding: 7bit 7, 25 -- X-Virus-Scanned: ClamAV 0.88.6/2269/Fri Dec 1 10:17:05 2006 on mta2 7, 25 -- X-Virus-Status: Clean ==============================End of - Headers==============================
In the UK 'school boards' offer nothing in the way of actual physical assistence with teaching - I suppose they may recommend specific slide sets if asked though. UK schools and teachers use their own budget and time to prepare lesson aids and buy science equipment. All the UK schools I have taught in had years-old boxes of prepared slides lying about (they keep for many years if prepared properly and not dropped or crushed under an objective by enthusiastic pupils) - so we never had to buy any new specimens. Lots of prepared slides are bit of snooze for under 16s though - a text book/internet search does it quicker and cheaper. Also try growing crystals on the slides as well.
Our lab technicians dealt with all the ordering anyway, we just had to make a general request for the item. You can also buy on ebay but the that is probably more for collectors of science equipment and quality may vary. However in the UK there is a lot of specialised suppliers of scientific equipment for secondary schools, who supply prepared slides. A small selection is:
I'm sure there's something linked in http://www.microscopy-uk.org.uk/
Plus for general microscope interest:
http://www.ukge.co.uk/UK/about.asp (for geological microscopes etc) http://science.nhmccd.edu/biol/slides.html a few pictures of slides + links http://micro.magnet.fsu.edu/ all you need to know about microscopes http://www.101science.com/Microscope.htm http://www.btinternet.com/~stephen.durr/ http://www.mccroneatlas.com http://www.diatoms.co.uk (just fun slides of diatoms and butteryfly scales)
I imagine there are school suppliers nearer to Cyprus, e.g. Greece or Turkey, but postage to Europe from the UK is cheap.
regards
Keith
--------------------------------------------
Dr Keith J Morris Manager Imaging Facilties Cell Biology Division The institute of Ophthalmology 11-43 Bath Street London EC1V 9EL
----- Original Message ----- X-from: {blegge-at-cwisp.ca} To: {keith.morris-at-ucl.ac.uk} Sent: Tuesday, November 28, 2006 7:08 PM
Debbie, Bill, et al
Been a couple of years since I've run an acrylamide gel, either for protein or sequencing. We fixed the gels in 30% Isopropanol/10%Acetic Acid prior to staining with coomassie blue.Destaining involves Acetone and Methanol, but i do not remember the precise information on the destaining solutions. After staining we destained with 16.5%Methanol/7.5% Acetic Acid. For sequencing I used 7% Methanol/5% Acetic acid. My memory is that some people used a formaldehyde fixation for sequencing gels, but I never tried that so do not know how well it would work.
Regards cryofixation/sectioning, you could do that, but it might be like using a 100megaton thermonuclear device to remove a tree stump. Remember that acrylamide is a plastic. Standard dehydration and embedding would work quite well as long as the target beads are held in place. Caveat, have never done it, so really do not know whether Bill or I am right. Maybe you do need the bomb.
This does not fit in our normal formaldehyde/glutaraldehyde fixation but it is consistent with fixation done for IF microscopy
==============================Original Headers============================== 4, 21 -- From paul_hazelton-at-umanitoba.ca Fri Dec 1 19:17:31 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 kB21HV28008406 4, 21 -- for {microscopy-at-microscopy.com} ; Fri, 1 Dec 2006 19:17:31 -0600 4, 21 -- Received: from [130.179.152.89] (cvx-026.cc.umanitoba.ca [130.179.152.89]) 4, 21 -- (authenticated bits=0) 4, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id kB21HQeb006824; 4, 21 -- Fri, 1 Dec 2006 19:17:27 -0600 (CST) 4, 21 -- Message-ID: {4570D421.50609-at-umanitoba.ca} 4, 21 -- Date: Fri, 01 Dec 2006 19:17:21 -0600 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: dsherman-at-purdue.edu, Microscopy Listserver {microscopy-at-microscopy.com} , 4, 21 -- Bill Tivol {tivol-at-caltech.edu} 4, 21 -- Subject: Re: [Microscopy] Embedding acrylamide gel 4, 21 -- References: {200612011945.kB1Jjmg0006334-at-ns.microscopy.com} 4, 21 -- In-Reply-To: {200612011945.kB1Jjmg0006334-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==============================
There are several suppliers in the US that can satisfy probably all of your needs.
One is Ward's Scientific and the other is Carolina Biological. Their slides range from $3US to about $15US depending on subject.
gary g.
At 11:06 AM 11/28/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Fri Dec 1 19:28:55 2006 10, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kB21SsEs019214 10, 20 -- for {microscopy-at-microscopy.com} ; Fri, 1 Dec 2006 19:28:55 -0600 10, 20 -- Received: (qmail 30313 invoked from network); 1 Dec 2006 17:28:48 -0800 10, 20 -- Received: by simscan 1.1.0 ppid: 30277, pid: 30308, t: 0.1694s 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; 1 Dec 2006 17:28:47 -0800 10, 20 -- Message-Id: {7.0.1.0.2.20061201172643.025ecbc0-at-gaugler.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 20 -- Date: Fri, 01 Dec 2006 17:28:51 -0800 10, 20 -- To: blegge-at-cwisp.ca 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: source for OM sample slides 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200611281906.kASJ61Fq004292-at-ns.microscopy.com} 10, 20 -- References: {200611281906.kASJ61Fq004292-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Mary, That might work, but one problem is that sulfur likes metal surfaces and it is very difficult to get it off. There was some work on it at Sandia National Lab in the early 80's that I am familiar with.
John, if you go this route, you might want to heat the nitrogen before going into the system. Run your N2 through a coiled tubing with heat tape on it. That will help strip the sulfur from the surface. You just want to make sure that you are in the viscous flow range which Mary's suggestion will do. Doing both her suggestion and mine together with the copper getter in the system might be a good way to get the sulfur out. I think that you will have to go longer than her suggested hour, though, probably overnight. Sulfur really likes those surfaces and has a high heat of absorption.
-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: mager-at-interchange.ubc.ca [mailto:mager-at-interchange.ubc.ca] Sent: Friday, December 01, 2006 2:52 PM To: Walck-at-SouthBayTech.com
Dear Dr. Mardinly, I would suspect that the rubber in the seal that failed is the source of the sulphur and this will react with the copper strongly. Unfortunately, this will be distributed throughout the system by the oil back-streamed by the system failure and can only be removed by very thorough cleaning of all chambers and lines. You might try baking the system after cleaning with solvents to get rig of all the oil. Change the oil in the backing pump and run everything for a week to clean it up. The other thing I have used successfully to clean up an SEM that was contaminating badly was to bleed in a stream of nitrogen gas through a feed through I made with a bit of fine stainless steel tubing and a metering valve. The gas valve was turned on until the SEM baffle valve closed and a bit more. Leave for about one hour. Take a blank port cover, drill a small hole through, glue the SS tubing through the hole, swage the fine metering valve on to the SS tubing. Attach the other side of the metering valve to a bag you can fill with nitrogen gas. A bag fill will last more than an hour at the right gas flow rate.) You can use a freshly-polished piece of copper to test your system. Good luck, Mary Mager Electron Microscopist Materials Eng. UBC #419 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA Phone: 604-822-5648 Fax: 604-822-3619 email: mager-at-interchange.ubc.ca
-----Original Message----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, December 01, 2006 11:42 AM To: mager-at-interchange.ubc.ca
We have a Gatan Model 655 Dry Pumping Station (www.gatan.com/holders/655_drypumpingstation.html) equipped with pods for storage of TEM specimens. In particular, TEM cross-sections of integrated circuits using copper interconnects seem to be particularly vulnerable to corrosion in atmosphere. This vacuum station has been essential in protecting the specimens that we put so much work into making, and has been very successful for quite some time. The dry vacuum system consists of a 'drag' pump, which is essentially a turbo pump, backed by a diaphragm type rough pump. Last summer, one of the rubber diaphragms failed in the backing pump, and the system ran for a few days with degraded vacuum. After the diaphragms were replaced and the high vacuum restored to the E-5 Torr range, we found that TEM cross-sections of integrated circuits using copper interconnects were destroyed in just a few hours if stored in this vacuum system. EDX of the copper corrosion product showed the presence of sulfur. We have tried cleaning the storage pods, and part of the inside of the vacuum chamber, but have been totally frustrated in being able to restore the essential function of this vacuum storage system. Any suggestions as to what may have happened, and what we can do to clean up the system would be very much appreciated. Thank you.
John Mardinly Intel Corporation
Disclaimer: This is a communication from this author and does not represent an opinion of Intel Corporation.
==============================Original Headers============================== 6, 30 -- From john.mardinly-at-intel.com Fri Dec 1 13:35:49 2006 6, 30 -- Received: from mga01.intel.com (mga01.intel.com [192.55.52.88]) 6, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB1JZnHb024448 6, 30 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 13:35:49 -0600 6, 30 -- Received: from fmsmga001.fm.intel.com ([10.253.24.23]) 6, 30 -- by mga01.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- Received: from scsmsx332.sc.intel.com (HELO scsmsx332.amr.corp.intel.com) ([10.3.90.6]) 6, 30 -- by fmsmga001.fm.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- X-ExtLoop1: 1 6, 30 -- X-IronPort-AV: i="4.09,486,1157353200"; 6, 30 -- d="scan'208"; a="171762165:sNHT19309206" 6, 30 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 30 -- Fri, 1 Dec 2006 11:35:31 -0800 6, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 30 -- Content-class: urn:content-classes:message 6, 30 -- MIME-Version: 1.0 6, 30 -- Content-Type: text/plain; 6, 30 -- charset="us-ascii" 6, 30 -- Subject: Contamination of Vacuum Specimen Storage System 6, 30 --
Debby- Polyacrylamide is one of the few plastics I have never found a way to section. Things I would try are: 1. lower the temperature to cryosection it (maybe below -100 C, which works for rubber) 2. embed the sample just like a biological tissue.
It would be surprising if you saw much, anyway. Proteins in a gel will make a hairpin shape due to SDS binding. Knowledge of the sample would help you to know what to expect. Carol
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==============================Original Headers============================== 4, 18 -- From heckman-at-bgnet.bgsu.edu Sat Dec 2 13:51:29 2006 4, 18 -- Received: from smtp01.bgsu.edu (smtp.bgsu.edu [129.1.5.17]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB2JpT4G027372 4, 18 -- for {microscopy-at-microscopy.com} ; Sat, 2 Dec 2006 13:51:29 -0600 4, 18 -- Received: from [129.1.85.81] (dhcp-85-81.bgsu.edu [129.1.85.81]) 4, 18 -- by smtp01.bgsu.edu (Switch-3.1.8/Switch-3.1.6) with ESMTP id kB2JpHrH003372; 4, 18 -- Sat, 2 Dec 2006 14:51:18 -0500 (EST) 4, 18 -- Mime-Version: 1.0 4, 18 -- X-Sender: heckman-at-mailstore.bgsu.edu 4, 18 -- Message-Id: {p04320403c197b32d3f44-at-[129.1.85.81]} 4, 18 -- In-Reply-To: {200612011944.kB1JiaXS004163-at-ns.microscopy.com} 4, 18 -- References: {200612011944.kB1JiaXS004163-at-ns.microscopy.com} 4, 18 -- Date: Sat, 2 Dec 2006 14:51:15 -0800 4, 18 -- To: dsherman-at-purdue.edu 4, 18 -- From: Carol Heckman {heckman-at-bgnet.bgsu.edu} 4, 18 -- Subject: Re: [Microscopy] Embedding acrylamide gel 4, 18 -- Cc: "Microscopy Listserver" {microscopy-at-microscopy.com} 4, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
} } (We apologise for any multiple postings) } } ************************************************************************ } } Dear friends and colleagues, } } Following numerous requests from prospective authors, the deadline for } submissions of papers to the 15th Scandinavian Conference on Image } Analysis 2007 (SCIA2007) has been extended. } } In order to get an early start on the process of distributing papers to } reviewers we have introduced two deadlines: } } 1) an *ABSTRACT & TITLE* deadline on December 8, 2006 } and } 2) a *PAPER* deadline on December 15th, 2006. } } NOTE: only papers with registered abstracts can be submitted after the } abstract & title deadline. Please note that no additional extension will } be granted. } } For more information on the conference and paper submission see } http://www.scia2007.dk } } We look forward to meeting you in Aalborg, Denmark in June 2007. } } Best regards, } } Kim Steenstrup Pedersen } Program chair for SCIA2007 } } -- } Ph.D., Cand. Scient. Kim Steenstrup Pedersen } Adjunkt (Assistant professor) } IT University of Copenhagen } Image Group } Rued Langgaardsvej 7, DK-2300 Copenhagen S, Denmark } Direct phone: +45 72 18 50 79 Fax: +45 72 18 50 01 } E-mail: kimstp-at-itu.dk } Home page: http://www.itu.dk/people/kimstp/ } } }
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Email: cjlcwright-at-wildblue.net Name: Christopher Wright
Title-Subject: [Filtered] Leo32 Windows 98 Control Software
Question: Hi,
I am trying to help a customer with an older Zeiss microscope. The issue is with the Leo32 software (I am actually a software techie, not a microscope tech.)
He had someone supporting him in the past, but they never gave him a copy of the Leo32 software, nor the administrator password into the software. Of course, they are not returning his calls.
So now that he is having all these problems, I am limited in what I can accomplish.
Is there any where I can get a hold of the original software or any easy way to "hack" the administrator password? (This is with the microscope owner's permission of course.)
He is several hours away, so I have only had a little time to look at this. I thought everything was working so I focused on getting good backups of the software, but he is calling now and it is still giving problems, so my backups are worthless. I just need to start over, or hack into the administrator setup.
I quite share this opinion, however it really depends on the kind of information this person needs. I don't think the gel morphology is of interest, but more the nanoparticles themselves. In this case one does not have to care too much about what happens to the gel right? It is only supposition here, I am afraid.
Perhaps one could just fix the gel as Paul described it and dry it with light heating and vacuum (like for protein gels). Perhaps it would possible to get some interesting image in SEM (BSE should show a big contrast between the gel and the particles), but perhaps the resolution would be an issue (and depth too). An alternative would be to embed the dried gel in resin (if it is possible, I have no idea but you'll know it only if you try it) and cut it for TEM observation.
Stephane
--- paul_hazelton-at-umanitoba.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 } ---------------------------------------------------------------------------- } } Debbie, Bill, et al } } Been a couple of years since I've run an acrylamide } gel, either for } protein or sequencing. We fixed the gels in 30% } Isopropanol/10%Acetic } Acid prior to staining with coomassie } blue.Destaining involves Acetone } and Methanol, but i do not remember the precise } information on the } destaining solutions. After staining we destained } with } 16.5%Methanol/7.5% Acetic Acid. For sequencing I } used 7% Methanol/5% } Acetic acid. My memory is that some people used a } formaldehyde fixation } for sequencing gels, but I never tried that so do } not know how well it } would work. } } Regards cryofixation/sectioning, you could do that, } but it might be like } using a 100megaton thermonuclear device to remove a } tree stump. } Remember that acrylamide is a plastic. Standard } dehydration and } embedding would work quite well as long as the } target beads are held in } place. Caveat, have never done it, so really do not } know whether Bill } or I am right. Maybe you do need the bomb. } } This does not fit in our normal } formaldehyde/glutaraldehyde fixation but } it is consistent with fixation done for IF } microscopy } } ==============================Original } Headers============================== } 4, 21 -- From paul_hazelton-at-umanitoba.ca Fri Dec 1 } 19:17:31 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 } kB21HV28008406 } 4, 21 -- for {microscopy-at-microscopy.com} ; Fri, 1 } Dec 2006 19:17:31 -0600 } 4, 21 -- Received: from [130.179.152.89] } (cvx-026.cc.umanitoba.ca [130.179.152.89]) } 4, 21 -- (authenticated bits=0) } 4, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) } with ESMTP id kB21HQeb006824; } 4, 21 -- Fri, 1 Dec 2006 19:17:27 -0600 (CST) } 4, 21 -- Message-ID: {4570D421.50609-at-umanitoba.ca} } 4, 21 -- Date: Fri, 01 Dec 2006 19:17:21 -0600 } 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: dsherman-at-purdue.edu, Microscopy } Listserver {microscopy-at-microscopy.com} , } 4, 21 -- Bill Tivol {tivol-at-caltech.edu} } 4, 21 -- Subject: Re: [Microscopy] Embedding } acrylamide gel } 4, 21 -- References: } {200612011945.kB1Jjmg0006334-at-ns.microscopy.com} } 4, 21 -- In-Reply-To: } {200612011945.kB1Jjmg0006334-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============================== }
____________________________________________________________________________________ Do you Yahoo!? Everyone is raving about the all-new Yahoo! Mail beta. http://new.mail.yahoo.com
==============================Original Headers============================== 11, 20 -- From nizets2-at-yahoo.com Mon Dec 4 07:26:48 2006 11, 20 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.91.144]) 11, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kB4DQlti028778 11, 20 -- for {microscopy-at-microscopy.com} ; Mon, 4 Dec 2006 07:26:48 -0600 11, 20 -- Received: (qmail 35682 invoked by uid 60001); 4 Dec 2006 13:26:47 -0000 11, 20 -- Message-ID: {20061204132647.35680.qmail-at-web37412.mail.mud.yahoo.com} 11, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 20 -- s=s1024; d=yahoo.com; 11, 20 -- h=X-YMail-OSG:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding:Message-ID; 11, 20 -- b=AhInLYdcrjbZSTlCY/HP9I2NGTp2/upI1A5ByIr9XD+zbzP+Py1Y6W4rbHeJ7cNcBpSpCMF6uEPOwI/+scIJrqh+KOn5sRviYHiDnzGdjI36FOK6dE7bOeM0A2RlEVZj2Brkx5aaZpsozsJBc8DFnAdKhH8hQqLOniIzvyLVE5g=; 11, 20 -- X-YMail-OSG: j16WOLwVM1m4WwPBeVNIoFEbHe9FRwcB0JDJ9bHUcADDINulx4fHT0ICnEksPPX4QdK3hygATNt2Ywlr60_5xURHARjlfIhbfoM2Y8L_N.VuABBvWoPmsKz.7zF6s8PXtdsUYEkZ813kB1aEB0qmdY1t_k.TQyJ.liIVQtogPLTjGEXS1qosYUplUCYBDTXYa9pKj8c6 11, 20 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Mon, 04 Dec 2006 05:26:47 PST 11, 20 -- Date: Mon, 4 Dec 2006 05:26:47 -0800 (PST) 11, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 11, 20 -- Subject: Re: [Microscopy] Re: Embedding acrylamide gel 11, 20 -- To: microscopy-at-microscopy.com 11, 20 -- In-Reply-To: {200612020121.kB21LZQD016441-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==============================
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The Analytical & Engineering Science Division of Hercules Incorporated has an immediate opening for an experimental microscopist with a strong background in structural and morphological characterization of materials to join the R&D team at our Corporate Research Center in Wilmington, DE.
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As with all Hercules employment practices, this process will comply with all applicable equal employment laws. No candidate will be discriminated against on the basis of race, sex, national origin, color, religion, veteran status, disability, or any other protected class.
Qualified candidates are encouraged to send an application letter and a detailed resume to Hercules Incorporated, Research Center, Attn: Human Resources, 500 Hercules Road, Wilmington, DE 19808-1599 or technology-at-herc.com. Refer to position Microscopist.
==============================Original Headers============================== 13, 24 -- From cni-at-udel.edu Mon Dec 4 08:19:59 2006 13, 24 -- Received: from md4.nss.udel.edu (md4.nss.udel.edu [128.175.1.14]) 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB4EJwOs008057 13, 24 -- for {Microscopy-at-microscopy.com} ; Mon, 4 Dec 2006 08:19:59 -0600 13, 24 -- Received: from Suba (em076-119.engg.udel.edu [128.175.119.76]) 13, 24 -- by md4.nss.udel.edu (MOS 3.8.2-GA) 13, 24 -- with ESMTP id DPS84448; 13, 24 -- Mon, 4 Dec 2006 09:19:58 -0500 (EST) 13, 24 -- From: "Chaoying Ni" {cni-at-udel.edu} 13, 24 -- To: {Microscopy-at-microscopy.com} 13, 24 -- Subject: Microscopist position at Hercules Incorporated 13, 24 -- Date: Mon, 4 Dec 2006 09:19:58 -0500 13, 24 -- Organization: University of Delaware 13, 24 -- Message-ID: {002e01c717af$475c85b0$4c77af80-at-Suba} 13, 24 -- MIME-Version: 1.0 13, 24 -- Content-Type: text/plain; 13, 24 -- charset="us-ascii" 13, 24 -- X-Priority: 3 (Normal) 13, 24 -- X-MSMail-Priority: Normal 13, 24 -- X-Mailer: Microsoft Outlook, Build 10.0.6626 13, 24 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 13, 24 -- Importance: Normal 13, 24 -- Content-Transfer-Encoding: 8bit 13, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kB4EJwOs008057 ==============================End of - Headers==============================
Dear John, One other possibility would be the XEI Scientific, Inc. decontaminator. It uses oxygen to clean your system and sample and might be more vigorous than the nitrogen at removing the sulphur residue. I have no affiliation with XEI, nor do I own one. Regards, Mary Mager
-----Original Message----- X-from: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] Sent: Friday, December 01, 2006 11:42 AM To: mager-at-interchange.ubc.ca
We have a Gatan Model 655 Dry Pumping Station (www.gatan.com/holders/655_drypumpingstation.html) equipped with pods for storage of TEM specimens. In particular, TEM cross-sections of integrated circuits using copper interconnects seem to be particularly vulnerable to corrosion in atmosphere. This vacuum station has been essential in protecting the specimens that we put so much work into making, and has been very successful for quite some time. The dry vacuum system consists of a 'drag' pump, which is essentially a turbo pump, backed by a diaphragm type rough pump. Last summer, one of the rubber diaphragms failed in the backing pump, and the system ran for a few days with degraded vacuum. After the diaphragms were replaced and the high vacuum restored to the E-5 Torr range, we found that TEM cross-sections of integrated circuits using copper interconnects were destroyed in just a few hours if stored in this vacuum system. EDX of the copper corrosion product showed the presence of sulfur. We have tried cleaning the storage pods, and part of the inside of the vacuum chamber, but have been totally frustrated in being able to restore the essential function of this vacuum storage system. Any suggestions as to what may have happened, and what we can do to clean up the system would be very much appreciated. Thank you.
John Mardinly Intel Corporation
Disclaimer: This is a communication from this author and does not represent an opinion of Intel Corporation.
==============================Original Headers============================== 6, 30 -- From john.mardinly-at-intel.com Fri Dec 1 13:35:49 2006 6, 30 -- Received: from mga01.intel.com (mga01.intel.com [192.55.52.88]) 6, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB1JZnHb024448 6, 30 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 13:35:49 -0600 6, 30 -- Received: from fmsmga001.fm.intel.com ([10.253.24.23]) 6, 30 -- by mga01.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- Received: from scsmsx332.sc.intel.com (HELO scsmsx332.amr.corp.intel.com) ([10.3.90.6]) 6, 30 -- by fmsmga001.fm.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 6, 30 -- X-ExtLoop1: 1 6, 30 -- X-IronPort-AV: i="4.09,486,1157353200"; 6, 30 -- d="scan'208"; a="171762165:sNHT19309206" 6, 30 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); 6, 30 -- Fri, 1 Dec 2006 11:35:31 -0800 6, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 30 -- Content-class: urn:content-classes:message 6, 30 -- MIME-Version: 1.0 6, 30 -- Content-Type: text/plain; 6, 30 -- charset="us-ascii" 6, 30 -- Subject: Contamination of Vacuum Specimen Storage System 6, 30 --
Advances in Electron Crystallography of Membrane Proteins: Call for papers for a special issue of JSB Ben Hankamer, Henning Stahlberg and Bob Glaeser, Guest Editors
The Journal of Structural Biology invites submissions for a special issue focusing on recent ‘Advances in Electron Crystallography of Membrane Proteins’. Electron-crystallography offers many advantages for the determination of membrane proteins structures, which is one of the great challenges of structural and cell biology.
In recent years electron-crystallography of membrane proteins has undergone major developments, which enhance its natural advantages. These advantages include the fact that membrane proteins can be crystallized within a near-native lipid bilayer environment, that 2-D crystals may sometimes be obtained even though 3-D crystals have not been, and that the technique is well suited for the crystallization of small and hydrophobic integral membrane proteins as well as membrane associated proteins. Functionalized monolayer technology is also opening up the possibility of streamlining crystal production and of coupling this process to improved protein expression techniques. As a resolution of better than 2Å has now been reported, it is proven that data can be collected to determine not only protein structure but its detailed interplay with the components of the lipid bilayer.
This special edition aims to capture the key developments that will help to take the electron crystallography field to the next phase, from streamlined crystal production and structure determination to new levels of resolution. Manuscripts in the following areas are specifically (but not exclusively) invited:
1. Membrane protein expression 2. Membrane protein purification 3. 2D crystallization 4. Sample preparation for electron microscopy 5. Automated screening of crystallization trials 6. Computer image processing and software developments 7. Recent applications and new structures
Papers can be in the form of research articles or mini-reviews that summarize key areas of importance. Prior to submission of a minireview, or if you require any further information on the suitability of your manuscript for inclusion in this special issue, please contact Ben Hankamer (b.hankamer-at-imb.uq.edu.au), Henning Stahlberg (HStahlberg-at-ucdavis.edu), or Bob Glaeser (RMGlaeser-at-lbl.gov) .
A brief e-mail by 22 Dec 2006, stating your intent to submit a contribution would be greatly appreciated as it would help us to plan the format of the special issue. The deadline for receipt of manuscripts for the Special Issue is 1 May 2006. Manuscripts will be peer-reviewed using the criteria of the Journal of Structural Biology (see http://www.academicpress.com/jsb). Submissions should be made on-line via the normal JSB submission process. In the cover letter please state that you wish the article to be considered for the Special Issue on ‘Advances in Electron Crystallography’ and mark for the attention of Ben Hankamer, Henning Stahlberg or Bob Glaeser.
_____________________________________________________________ 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 http://2dx.org _____________________________________________________________
==============================Original Headers============================== 17, 21 -- From HStahlberg-at-ucdavis.edu Mon Dec 4 12:47:54 2006 17, 21 -- Received: from warsaw.ucdavis.edu (warsaw.ucdavis.edu [128.120.32.41]) 17, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB4IlrhL006058 17, 21 -- for {Microscopy-at-microscopy.com} ; Mon, 4 Dec 2006 12:47:53 -0600 17, 21 -- Received: from [169.237.214.65] ([169.237.214.65]) 17, 21 -- by warsaw.ucdavis.edu (8.13.7/8.13.1/it-defang-5.4.0) with ESMTP id kB4IlJdD026384; 17, 21 -- Mon, 4 Dec 2006 10:47:19 -0800 (PST) 17, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 17, 21 -- Content-Type: text/plain; charset=WINDOWS-1252; delsp=yes; format=flowed 17, 21 -- Message-Id: {FAD32BC8-BB32-48DE-80D6-32673C0C00B5-at-ucdavis.edu} 17, 21 -- Cc: Ben Hankamer {b.hankamer-at-imb.uq.edu.au} , 17, 21 -- Henning Stahlberg {HStahlberg-at-ucdavis.edu} , 17, 21 -- "Robert M. Glaeser" {RMGlaeser-at-lbl.gov} 17, 21 -- From: Henning Stahlberg {HStahlberg-at-ucdavis.edu} 17, 21 -- Subject: Advances in Electron Crystallography of Membrane Proteins: Call for papers for a special issue of JSB 17, 21 -- Date: Mon, 4 Dec 2006 10:47:18 -0800 17, 21 -- To: Microscopy-at-microscopy.com 17, 21 -- X-Mailer: Apple Mail (2.752.2) 17, 21 -- X-Scanned-By: MIMEDefang 2.57 on 128.120.32.41 17, 21 -- Content-Transfer-Encoding: 8bit 17, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kB4IlrhL006058 ==============================End of - Headers==============================
Many thanks for all the replies. Your suggestions and experiences give me some more things to think about and run past the service engineers and users (who are being very cooperative).
I should have mentioned in my original post that this system is dry pumped so pumping oil is not in the picture (I don't know why I did not think to mention that since that would have been one of my first questions).
Several of you have suggested that I have the contamination analyzed with FTIR and I am in the process of arranging that.
Again, many thanks and I will let you know about any thing that comes to light.
Tom
Tom Stephens Assistant Research Scientist Microscopy and Imaging Center Texas A&M University College Station, TX 77843 office phone:979-845-1165 email:tstephen-at-mic.tamu.edu
==============================Original Headers============================== 8, 20 -- From tstephen-at-mic.tamu.edu Mon Dec 4 15:17:57 2006 8, 20 -- Received: from sr-5-int.cis.tamu.edu (smtp-relay.tamu.edu [165.91.22.120]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB4LHv2n020145 8, 20 -- for {Microscopy-at-Microscopy.Com} ; Mon, 4 Dec 2006 15:17:57 -0600 8, 20 -- Received: from localhost (localhost.tamu.edu [127.0.0.1]) 8, 20 -- by sr-5-int.cis.tamu.edu (Postfix) with ESMTP id 5D27F70EC 8, 20 -- for {Microscopy-at-Microscopy.Com} ; Mon, 4 Dec 2006 15:17:57 -0600 (CST) 8, 20 -- Received: from jntcspc.mic.tamu.edu (cyn6400.tamu.edu [165.91.109.143]) 8, 20 -- by sr-5-int.cis.tamu.edu (Postfix) with ESMTP id 9DEEC70E2 8, 20 -- for {Microscopy-at-Microscopy.Com} ; Mon, 4 Dec 2006 15:17:56 -0600 (CST) 8, 20 -- Message-Id: {5.2.0.9.0.20061201122251.02a969f0-at-mic.tamu.edu} 8, 20 -- X-Sender: tstephen-at-mic.tamu.edu (Unverified) 8, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.0.9 8, 20 -- Date: Mon, 04 Dec 2006 15:18:06 -0600 8, 20 -- To: Microscopy-at-Microscopy.Com 8, 20 -- From: Tom Stephens {tstephen-at-mic.tamu.edu} 8, 20 -- Subject: Re: Re: [Microscopy] SEM - Mystery contamination of FESEM 8, 20 -- Mime-Version: 1.0 8, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 8, 20 -- X-Virus-Scanned: amavisd-new at tamu.edu ==============================End of - Headers==============================
Hello, I have noticed strange behavior of the Emission Meter of our Philips CM100 electron microscope. When the microscope is ON with HT switched OFF, the pointer of the meter is in the leftmost position indicating negative current!? It does not change even when HT is switched ON (40kV, 60kV, 80kV, 100kV). No change can also be observed when the filament is heated; the pointer stuck in leftmost position. However, when the microscope is in STAND BY, the pointer is in normal position indicating to zero.
I have changed the whenelt for the other one with a new cathode, but it did not help. Does anybody has an explanation for this? Thanking you in advance for any suggestion.
Oldrich
------------------------------------------ Oldrich Benada Institute of Microbiology Acad. Sci. CR Videnska 1083 CZ-142 20 Prague 4 Czech Republic
==============================Original Headers============================== 4, 20 -- From benada-at-biomed.cas.cz Tue Dec 5 08:20:58 2006 4, 20 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB5EKvAF021183 4, 20 -- for {microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 08:20:58 -0600 4, 20 -- Received: from [147.231.44.101] (u117ob.mbu.cas.cz [147.231.44.101]) 4, 20 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id kB5EJvVf008075 4, 20 -- for {microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 15:20:01 +0100 (CET) 4, 20 -- From: benada-at-biomed.cas.cz 4, 20 -- To: microscopy-at-microscopy.com 4, 20 -- Date: Tue, 05 Dec 2006 15:20:48 +0100 4, 20 -- MIME-Version: 1.0 4, 20 -- Subject: Emission Meter - Philips CM100 4, 20 -- Message-ID: {45758E50.10737.568DD6-at-benada.biomed.cas.cz} 4, 20 -- Priority: normal 4, 20 -- X-mailer: Pegasus Mail for Windows (4.41) 4, 20 -- Content-type: text/plain; charset=US-ASCII 4, 20 -- Content-transfer-encoding: 7BIT 4, 20 -- Content-description: Mail message body 4, 20 -- X-Antivirus: avast! (VPS 0653-5, 05.12.2006), Outbound message 4, 20 -- X-Antivirus-Status: Clean ==============================End of - Headers==============================
I'm having a weird problem with the LN2 dipstick on my EDS detector. Background - this is a PGT Avalon system that uses an interface box between the dipstick and the Avalon electronics. The dipstick has a 1K ohm resister in the tip and a potentiometer in the interface box is used to set the low LN2 point. The low LN2 point is set correctly and when the thing is working correctly, removing the stick from LN2 will shut off the HV.
Here is the problem. When I drop the dipstick into the EDS detector dewar (full of LN2) the ratemeter on the Avalon electronics pegs to 100% deadtime. If I take it out the ratemeter the deadtime goes back to near 0. Repeating yields same results. Here is the mind bender - if I drop the dipstick into a vacuum flask full of LN2 the ratemeter stays at ~0% deadtime.
I have done the following;
Bypassed the dipstick and cabling by jumpering a 1k ohm resistor directly into the interface box. I drop the resistor into LN2 and I see no noise in ratemeter. Next I added back in the cable with the resistor, dip it into the dewar flask and it works fine. Next I add the dipstick and cable and they work fine. Next I drop the dipstick ( connected to interface box by cable) into the detector dewar and the ratemeter pegs 100% DT.
I've rebuilt the dipstick - new resistor, and wiring. New cable from dipstick to interface box. Still same problem.
Any ideas??
Owen Mills Michigan Tech University
} all done w/ no ebeam. } } 1. with dipstick in detector dewar DT=100%. } 2. remove dipstick and DT drops to ~0. } 3. this happens every time. } 4. drop dipstick into flask of ln2 - ~ 0% DT } } so what is different about the detector dewar? } } btw, if you drop a 1k ohm resistor into ln2 the resistance goes up } to 1.7k ohm. } } let me know if you can figure this out. } } Owen }
==============================Original Headers============================== 14, 31 -- From opmills-at-mtu.edu Tue Dec 5 09:04:09 2006 14, 31 -- Received: from node8.mtu.edu (node8.mtu.edu [141.219.69.8]) 14, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB5F49DS000647 14, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 09:04:09 -0600 14, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 14, 31 -- by node8.mtu.edu (8.13.1/8.13.1) with ESMTP id kB5F49Z1028389 14, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 10:04:09 -0500 14, 31 -- Received: from node23.edge.dcsint.mtu.edu (node23.mtu.edu [141.219.69.23]) 14, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id kB5F49qL013649 14, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 10:04:09 -0500 14, 31 -- Received: from mail.mtu.edu (campus0.mtu.edu [141.219.70.8]) 14, 31 -- by node23.edge.dcsint.mtu.edu (8.12.11.20060614/8.12.11) with ESMTP id kB5F485b021445 14, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 10:04:08 -0500 14, 31 -- (envelope-from opmills-at-mtu.edu) 14, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 14, 31 -- by mail.mtu.edu (8.13.6+Sun/8.11.6) with ESMTP id kB5F47MF010989 14, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 10:04:07 -0500 (EST) 14, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) 14, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id kB5F47GK013637 14, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 10:04:07 -0500 14, 31 -- Mime-Version: 1.0 (Apple Message framework v752.3) 14, 31 -- Content-Transfer-Encoding: 7bit 14, 31 -- Message-Id: {16BADD7E-DC55-4F45-80E3-EE2B827AD956-at-mtu.edu} 14, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 14, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 14, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} 14, 31 -- Subject: EDS dipstick conundrum 14, 31 -- Date: Tue, 5 Dec 2006 10:04:06 -0500 14, 31 -- X-Mailer: Apple Mail (2.752.3) 14, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2006.12.5.63432 14, 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==============================
If emission control still works, then troubleshoot emission meter circuit. It is hard to be more specific at this point. I don't think that cleaning and column alignment will help.
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: {benada-at-biomed.cas.cz} To: {vitalylazar-at-att.net} Sent: Tuesday, December 05, 2006 9:24 AM
Oldrich,
We have a Philips CM200 with similar issues with the emission meter. Our service engineer has told us that the emission meter has had a history of problems. His suggestion is to use CM Remote to read the emission current to an external PC.
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Hello, I have noticed strange behavior of the Emission Meter of our Philips CM100 electron microscope. When the microscope is ON with HT switched OFF, the pointer of the meter is in the leftmost position indicating negative current!? It does not change even when HT is switched ON (40kV, 60kV, 80kV, 100kV). No change can also be observed when the filament is heated; the pointer stuck in leftmost position. However, when the microscope is in STAND BY, the pointer is in normal position indicating to zero.
I have changed the whenelt for the other one with a new cathode, but it did not help. Does anybody has an explanation for this? Thanking you in advance for any suggestion.
Oldrich
------------------------------------------ Oldrich Benada Institute of Microbiology Acad. Sci. CR Videnska 1083 CZ-142 20 Prague 4 Czech Republic
==============================Original Headers============================== 12, 23 -- From david.r.hull-at-nasa.gov Tue Dec 5 12:19:06 2006 12, 23 -- Received: from ndjsvws04.ndc.nasa.gov (ndjsvws04.ndc.nasa.gov [198.120.25.84]) 12, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB5IJ5oo028662 12, 23 -- for {Microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 12:19:05 -0600 12, 23 -- Received: from ndjsxgw01.ndc.nasa.gov ([129.166.32.111]) by ndjsvws04.ndc.nasa.gov with InterScan Messaging Security Suite; Tue, 05 Dec 2006 12:19:04 -0600 12, 23 -- Received: from NDJSEVS16.ndc.nasa.gov ([129.166.32.126]) by ndjsxgw01.ndc.nasa.gov with Microsoft SMTPSVC(6.0.3790.1830); 12, 23 -- Tue, 5 Dec 2006 12:19:02 -0600 12, 23 -- Received: from 129.166.32.15 ([129.166.32.15]) by NDJSEVS16.ndc.nasa.gov ([129.166.32.136]) via Exchange Front-End Server mail01.ndc.nasa.gov ([129.166.32.105]) with Microsoft Exchange Server HTTP-DAV ; 12, 23 -- Tue, 5 Dec 2006 18:19:02 +0000 12, 23 -- User-Agent: Microsoft-Entourage/11.2.5.060620 12, 23 -- Date: Tue, 05 Dec 2006 13:21:09 -0500 12, 23 -- Subject: FW: [Microscopy] Emission Meter - Philips CM100 12, 23 -- From: "Hull, David R" {David.R.Hull-at-nasa.gov} 12, 23 -- To: {Microscopy-at-microscopy.com} 12, 23 -- Message-ID: {C19B22C5.2127%David.R.Hull-at-nasa.gov} 12, 23 -- Thread-Topic: [Microscopy] Emission Meter - Philips CM100 12, 23 -- Thread-Index: AccYmiK6YWXUH4SNEduBcAAKldSx3A== 12, 23 -- In-Reply-To: {200612051422.kB5EMcGw022425-at-ns.microscopy.com} 12, 23 -- Mime-version: 1.0 12, 23 -- Content-type: text/plain; 12, 23 -- charset="US-ASCII" 12, 23 -- Content-transfer-encoding: 7bit 12, 23 -- X-OriginalArrivalTime: 05 Dec 2006 18:19:02.0417 (UTC) FILETIME=[D7471010:01C71899] ==============================End of - Headers==============================
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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
Earth loop causing noise pickup etc into your analyser? When you put the dipstick into the detector dewar you may be inadvertently closing the loop somehow? Look for exposed or broken earthing braid etc?
Arthur.
} } All, } } I'm having a weird problem with the LN2 dipstick on my EDS detector. } Background - this is a PGT Avalon system that uses an interface box } between the dipstick and the Avalon electronics. The dipstick has a } 1K ohm resister in the tip and a potentiometer in the interface box } is used to set the low LN2 point. The low LN2 point is set correctly } and when the thing is working correctly, removing the stick from LN2 } will shut off the HV. } } Here is the problem. When I drop the dipstick into the EDS detector } dewar (full of LN2) the ratemeter on the Avalon electronics pegs to } 100% deadtime. If I take it out the ratemeter the deadtime goes back } to near 0. Repeating yields same results. Here is the mind bender - } if I drop the dipstick into a vacuum flask full of LN2 the ratemeter } stays at ~0% deadtime. } } I have done the following; } } Bypassed the dipstick and cabling by jumpering a 1k ohm resistor } directly into the interface box. I drop the resistor into LN2 and I } see no noise in ratemeter. Next I added back in the cable with the } resistor, dip it into the dewar flask and it works fine. Next I add } the dipstick and cable and they work fine. Next I drop the dipstick } ( connected to interface box by cable) into the detector dewar and } the ratemeter pegs 100% DT. } } I've rebuilt the dipstick - new resistor, and wiring. New cable from } dipstick to interface box. Still same problem. } } Any ideas?? } } Owen Mills } Michigan Tech University } } } } } } } } all done w/ no ebeam. } } } } 1. with dipstick in detector dewar DT=100%. } } 2. remove dipstick and DT drops to ~0. } } 3. this happens every time. } } 4. drop dipstick into flask of ln2 - ~ 0% DT } } } } so what is different about the detector dewar? } } } } btw, if you drop a 1k ohm resistor into ln2 the resistance goes up } } to 1.7k ohm. } } } } let me know if you can figure this out. } } } } Owen } } }
==============================Original Headers============================== 5, 27 -- From ard-at-ansto.gov.au Tue Dec 5 19:01:12 2006 5, 27 -- Received: from tachyon.gw.ansto.gov.au (tachyon.gw.ansto.gov.au [137.157.8.253]) 5, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB611AMs028183 5, 27 -- for {microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 19:01:11 -0600 5, 27 -- Received: (from uucp-at-localhost) 5, 27 -- by tachyon.gw.ansto.gov.au (8.11.7p1+Sun/8.11.7) id kB6103o16961; 5, 27 -- Wed, 6 Dec 2006 12:00:03 +1100 (EST) 5, 27 -- Received: from jenner.ansto.gov.au(137.157.59.25) by tachyon.gw.ansto.gov.au via csmap (V6.0) 5, 27 -- id srcCAA5OaOeH; Wed, 6 Dec 06 12:00:03 +1100 5, 27 -- Received: from hadron.ansto.gov.au (hadron.ansto.gov.au [137.157.13.219]) 5, 27 -- by jenner.ansto.gov.au (8.12.10/8.12.10) with ESMTP id kB60umVr026928; 5, 27 -- Wed, 6 Dec 2006 11:56:48 +1100 5, 27 -- Received: from [137.157.95.82] (arthur.amat.ansto.gov.au [137.157.95.82]) 5, 27 -- by hadron.ansto.gov.au (8.9.3/8.9.3) with ESMTP id LAA26475; 5, 27 -- Wed, 6 Dec 2006 11:56:42 +1100 (EST) 5, 27 -- Mime-Version: 1.0 5, 27 -- X-Sender: ard-at-hadron.ansto.gov.au 5, 27 -- Message-Id: {v04210100c19bc495d087-at-[137.157.95.82]} 5, 27 -- In-Reply-To: {200612051505.kB5F54ks002572-at-ns.microscopy.com} 5, 27 -- References: {200612051505.kB5F54ks002572-at-ns.microscopy.com} 5, 27 -- Date: Wed, 6 Dec 2006 11:56:42 +1100 5, 27 -- To: {microscopy-at-microscopy.com} 5, 27 -- From: Arthur Day {ard-at-ansto.gov.au} 5, 27 -- Subject: Re: [Microscopy] EDS dipstick conundrum 5, 27 -- Cc: opmills-at-mtu.edu 5, 27 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 5, 27 -- X-ANSTO-MailScanner: Found to be clean ==============================End of - Headers==============================
There is a possibility the problem is not electrical. The detector may have become more microphonic (or more heat is being liberated by the dipstick).
In this scenario, the dipstick causes boiling of the LN2 and the vibration is causing the deadtime increase. To test this, simply insert a room temperature metal rod to the same depth (e.g. a welding rod or two).
If the DT increases, boiling vibes are likely the trouble. If not, it is probably a (somehow) ground loop.
Good luck & best regards,
Woody White BWXT Services
==============================Original Headers============================== 9, 26 -- From nwwhite-at-bwxt.com Wed Dec 6 07:03:26 2006 9, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 9, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB6D3PNV015277 9, 26 -- for {microscopy-at-msa.microscopy.com} ; Wed, 6 Dec 2006 07:03:26 -0600 9, 26 -- Received: from ([131.184.13.224]) 9, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.3073251; 9, 26 -- Wed, 06 Dec 2006 08:03:03 -0500 9, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 9, 26 -- Wed, 6 Dec 2006 08:03:03 -0500 9, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 26 -- Content-class: urn:content-classes:message 9, 26 -- MIME-Version: 1.0 9, 26 -- Content-Type: text/plain; 9, 26 -- charset="us-ascii" 9, 26 -- Subject: RE: [Microscopy] EDS dipstick conundrum 9, 26 -- Date: Wed, 6 Dec 2006 08:03:02 -0500 9, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D0E5-at-BWXSPO01.BWXS.BWXTECH.NET} 9, 26 -- X-MS-Has-Attach: 9, 26 -- X-MS-TNEF-Correlator: 9, 26 -- Thread-Topic: [Microscopy] EDS dipstick conundrum 9, 26 -- Thread-Index: AccYfrnYz830PGqvRPCOCPE8U2zxTgAtuHIg 9, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 9, 26 -- To: {opmills-at-mtu.edu} , "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 9, 26 -- X-OriginalArrivalTime: 06 Dec 2006 13:03:03.0316 (UTC) FILETIME=[DD2F9540:01C71936] 9, 26 -- Content-Transfer-Encoding: 8bit 9, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kB6D3PNV015277 ==============================End of - Headers==============================
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Email: vthawfeek-at-yahoo.co.in Name: V.Thawfeek Mohammed
Organization: psg tech coll. of engg.
Title-Subject: [Filtered] optical physics
Question: why does a light beam diverge when it strikes a medium with greater refractive index than the medium ,in which it is.
unfortunately I am neither a light beam nor a medium with greater or smaller refractive index than the other medium.....so I can't tell you (truly) why (:-)) ...but I think you should have in mind the "refraction law" of Snellius (Snel's - Snell's - Snellius' Law, refraction law, law of refractive index) which is defined as:
n times sin alpha = n' times sin.ß
(I try to translate a text from German, apologize if not all terms are correct "English" : a ray/light beam incident under a certain angle } alpha { (with regard to the area of entry = incidence perpendicle) to the border/boundary of two optic media with refraction index n and n' respectively, will be bent/refracted. The resultant beam will transit through the second medium with refractory index n' under a certain angle ß with regard to the incidence perpendicle).
search Google for either term of SNELLIUS (see above) or directly ==} http://en.wikipedia.org/wiki/Snell's_law Hope this helps best regards,
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Question: why does a light beam diverge when it strikes a medium with greater refractive index than the medium, in which it is.
Scott; Thanks to you and everyone that responded with their thoughts on this problem. The feeling among our group now is that your thought that outgassing from the grease in the mechanical part of the diaphragm pumps can backstream when the diaphragms fail is the most likely diagnosis. This particular pump had a huge tear in the failed diaphragm, but this was a bit of a surprise, since our many Gatan PIPS have exactly the same pumps, and we routinely run them until diaphragm failure, although they tend to fail with smaller tears, and perhaps the gas load from the argon ion guns tends to keep them cleaner by purging backstreaming. We have cleaned the vacuum chamber with solvents (and it was a mess), but cleaning the turbo pump will be difficult, since it cannot be disassembled. A nitrogen purge will probably be the best method of cleaning the turbo. We also now intend to proactively change all of our diaphragms on an annual basis, rather than running them to failure, in the hope that this sort of mess never recurs.
John Mardinly Intel Corporation
Disclaimer: This is a communication from this author and does not represent an opinion of Intel Corporation.
-----Original Message----- X-from: Scott Walck [mailto:walck-at-southbaytech.com] Sent: Friday, December 01, 2006 2:43 PM To: Microscopy-at-microscopy.com Cc: Mardinly, John
We have a Gatan Model 655 Dry Pumping Station (www.gatan.com/holders/655_drypumpingstation.html) equipped with pods for storage of TEM specimens. In particular, TEM cross-sections of integrated circuits using copper interconnects seem to be particularly vulnerable to corrosion in atmosphere. This vacuum station has been essential in protecting the specimens that we put so much work into making, and has been very successful for quite some time. The dry vacuum system consists of a 'drag' pump, which is essentially a turbo pump, backed by a diaphragm type rough pump. Last summer, one of the rubber diaphragms failed in the backing pump, and the system ran for a few days with degraded vacuum. After the diaphragms were replaced and the high vacuum restored to the E-5 Torr range, we found that TEM cross-sections of integrated circuits using copper interconnects were destroyed in just a few hours if stored in this vacuum system. EDX of the copper corrosion product showed the presence of sulfur. We have tried cleaning the storage pods, and part of the inside of the vacuum chamber, but have been totally frustrated in being able to restore the essential function of this vacuum storage system. Any suggestions as to what may have happened, and what we can do to clean up the system would be very much appreciated. Thank you.
John Mardinly Intel Corporation
Disclaimer: This is a communication from this author and does not represent an opinion of Intel Corporation.
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Electron Microscopy Specialist The Stowers Institute for Medical Research has an opening for an Electron Microscopy (EM) Specialist in the Histology Department. Responsibilities include providing high quality research EM services using established protocols; daily operation of the electron microscopy lab; using and maintaining the microscopes and ancillary equipment; assisting researchers with specimen preparation; sample preparation, including sample receipt, fixation, processing, and embedding; development of protocols; training SIMR research members and other users of techniques and equipment; development of seminars and/or presentations for internal training; purchasing supplies; and maintaining records/archives of the EM lab operation. In addition to excellent organizational, communication, and problem solving skills, the successful candidate should be familiar with operation of all applicable specimen preparation equipment and microscopes (TEM and SEM); skilled at standard specimen preparation protocols and identifying associated artifacts; have experience in high pressure freezing, cryoEM techniques, and immuno EM; be able to lift in excess of 30 pounds; and be able work overtime, including weekday, weekend, or on call work. The minimum requirements include an Associates Degree in a biological science and four years experience in research EM. Or a four year degree in a biological science and two years experience in research EM. Check out all our many benefits at www.stowers-institute.org. Rhonda Allen BA HT(ASCP)HTL, QIHC Histotechnology Specialist II Stowers Institute 1000 E. 50th Street Kansas City, MO 64110 816-926-4305
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Mohammed, If memory serves me, the bending occurs due to a change in the speed of light in different materials. A higher refractive index indicates a lower speed.
The speed of light that is immutable, according to Einstein, is the speed of light in a vacuum.
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: vthawfeek-at-yahoo.co.in [mailto:vthawfeek-at-yahoo.co.in] Sent: Wednesday, December 06, 2006 9:20 AM To: kenconverse-at-qualityimages.biz
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Email: vthawfeek-at-yahoo.co.in Name: V.Thawfeek Mohammed
Organization: psg tech coll. of engg.
Title-Subject: [Filtered] optical physics
Question: why does a light beam diverge when it strikes a medium with greater refractive index than the medium ,in which it is.
The answer is really quite simple: Snell's Law. The bending of light as it crosses the boundary from one RI to another is governed by the Laws of Refraction.
The bending discussed in your question requires that several conditions be met: First, that there be different RIs on each side of the boundary. Second, that the light approach the boundary at an angle.
To understand what happens, it is first necessary to understand that refractive index is actually a measure of the impact of interaction between the electric field of light and the electric field of matter. The greater the interaction, the more slowly light will travel through that material and the higher the refractive index. For instance: RI = velocity of light in air/velocity of light in material RI of air = 1, velocity of light = 300,000 km/s RI of water is 1.33, velocity of light is 225,000 km/s RI of immersion oil, glass, and many polymers is approximately 1.5, velocity of light = 200,000 km/sec.
As for refraction, the analogy which is often used is that of a person roller skating (or in this day and age, roller blading) from one surface to another. For example, from the concrete sidewalk onto grass. The smooth surface of the concrete is analogous to a material with low refractive index; the rougher surface of glass is analogous to a material with higher RI. If the skater approaches the sidewalk:grass boundary with both feet parallel, both feet will slow down by the same amount and the skater will continue skating in the same direction. However, if the skater approaches the boundary at an angle, one foot will slow down while the other remains at the original speed. That different in speed causes the skater to pivot toward the material of higher RI.
If you think of light in terms of a wave front rather than a simple ray, the analogy transfers easily. If the wave approaches a boundary at an angle, the edge of the front which hits the higher RI first will slow down, causing the whole wave front to pivot around that point. That's what happens, for example, when light travels from air (RI = 1.00) into a glass coverslip (RI ~1.5) or a droplet of water or a cell (RI ~1.33). You can use the optical axis of the microscope as a reference. In these cases, the light will bend in TOWARD the optical axis. Conversely, when light emerges from a glass slide or a droplet of water it will bend AWAY from the OA. Actually this is why we use oil immersion: to cause those rays which would normally bend away from the OA, causing a loss of both intensity and the critical contribution to resolution and edge definition, to bend back toward the OA, where they have an opportunity to be captured by the objective and make a positive contribution to better imaging.
All of this is explained in detail, with diagrams in Optimizing Light Microscopy. If you are interested in a copy, please contact Ken Piel here in the MME office for purchasing details (see below).
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 next April. 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 or email him at kenpiel-at-mme1.com
At 08:24 AM 12/6/2006, vthawfeek-at-yahoo.co.in wrote:
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-----Original Message----- X-from: kenconverse-at-qualityimages.biz [mailto:kenconverse-at-qualityimages.biz] Sent: Wednesday, December 06, 2006 10:07 To: Mike Bode
Mohammed, If memory serves me, the bending occurs due to a change in the speed of light in different materials. A higher refractive index indicates a lower speed.
The speed of light that is immutable, according to Einstein, is the speed of light in a vacuum.
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: vthawfeek-at-yahoo.co.in [mailto:vthawfeek-at-yahoo.co.in] Sent: Wednesday, December 06, 2006 9:20 AM To: kenconverse-at-qualityimages.biz
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Email: vthawfeek-at-yahoo.co.in Name: V.Thawfeek Mohammed
Organization: psg tech coll. of engg.
Title-Subject: [Filtered] optical physics
Question: why does a light beam diverge when it strikes a medium with greater refractive index than the medium ,in which it is.
You have gotten some very good answers about why you see a light beam changes direction, or refracts, when hitting a medium with a different refractive index.
In your question, you use the word diverge. I'm not certain how you are using that word. But perhaps you want to know why the light beam changes size. Why light spreads out into a larger width, divergence, then the answer to that question adds in another aspect of refraction.
As you can read from the earlier responses, the reason that light refracts, or changes course, is a function of it's wavelength. A light beam is composed of many different wavelengths, depending upon it's source. Each wavelength diffracts slightly differently.
This will give you the change in the "size" of the light beam.
dj
On Wed, 6 Dec 2006, vthawfeek-at-yahoo.co.in wrote:
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==============================Original Headers============================== 9, 21 -- From "dljones-at-bestweb.net"-at-bestweb.net Wed Dec 6 12:01:52 2006 9, 21 -- Received: from mta2.srv.hcvlny.cv.net (mta2.srv.hcvlny.cv.net [167.206.4.197]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB6I1qEn017043 9, 21 -- for {microscopy-at-microscopy.com} ; Wed, 6 Dec 2006 12:01:52 -0600 9, 21 -- Received: from localhost (ool-44c62883.dyn.optonline.net [68.198.40.131]) 9, 21 -- by mta2.srv.hcvlny.cv.net 9, 21 -- (Sun Java System Messaging Server 6.2-6.01 (built Apr 3 2006)) 9, 21 -- with ESMTP id {0J9V007LC6310XD0-at-mta2.srv.hcvlny.cv.net} for 9, 21 -- microscopy-at-microscopy.com; Wed, 06 Dec 2006 13:01:51 -0500 (EST) 9, 21 -- Date: Wed, 06 Dec 2006 12:50:06 -0500 (Eastern Standard Time) 9, 21 -- From: "David L. Jones" {"dljones-at-bestweb.net"-at-bestweb.net} 9, 21 -- Subject: Re: [Microscopy] viaWWW: optical physics 9, 21 -- In-reply-to: {200612061421.kB6EL8M9015801-at-ns.microscopy.com} 9, 21 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 9, 21 -- To: vthawfeek-at-yahoo.co.in 9, 21 -- Cc: dljones-at-bestweb.net, microscopy-at-microscopy.com 9, 21 -- Message-id: {Pine.WNT.4.64.0612061235110.3120-at-dljtoshiba} 9, 21 -- MIME-version: 1.0 9, 21 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 9, 21 -- Content-transfer-encoding: 7BIT 9, 21 -- References: {200612061421.kB6EL8M9015801-at-ns.microscopy.com} ==============================End of - Headers==============================
This is a really fine answer. I would add one more analogy, since the question (I think) relates to the change in direction of the path of the light beam. In my classes, I refer to the image of a column of a marching band going from a concrete surface to a broken field. If they are exactly perpendicular to the boundary, they continue to go straight, although with some stumbles. If they enter at an angle, then those members who are on the "acute" side of the column slow down first. This "pulls the others along as well, bit by bit, and the overall direction of the march changes.
Date sent: Wed, 6 Dec 2006 11:21:25 -0600 To: jbs-at-temple.edu X-from: bfoster-at-mme1.com Send reply to: bfoster-at-mme1.com
Greetings, These and other wonderful answers to the Snell law question have all been wave-based. Is there an equivalent photon-based answer? Presumably, if you fired one photon at a time at an oblique surface you would get the most counts at the Snell angle; but, with single photons, how are we to understand lines of coherent marchers or even two feet on roller blades? Is this light being weird?
Just wondering... Tobias
} This is a really fine answer. I would add one more analogy, since } the question (I think) relates to the change in direction of the path } of the light beam. In my classes, I refer to the image of a column } of a marching band going from a concrete surface to a broken field. } If they are exactly perpendicular to the boundary, they continue to } go straight, although with some stumbles. If they enter at an angle, } then those members who are on the "acute" side of the column slow } down first. This "pulls the others along as well, bit by bit, and } the overall direction of the march changes. } } Date sent: Wed, 6 Dec 2006 11:21:25 -0600 } To: jbs-at-temple.edu } X-from: bfoster-at-mme1.com } Send reply to: bfoster-at-mme1.com } Subject: [Microscopy] Re: viaWWW: optical physics } } } } } } } } } } } } Hi, } } } } The answer is really quite simple: Snell's Law. The bending of } } light as it crosses the boundary from one RI to another is governed } } by the Laws of Refraction. } } } } The bending discussed in your question requires that several } } conditions be met: First, that there be different RIs on each side } } of the boundary. Second, that the light approach the boundary at } } an angle. } } } } To understand what happens, it is first necessary to understand } } that refractive index is actually a measure of the impact of } } interaction between the electric field of light and the electric } } field of matter. The greater the interaction, the more slowly } } light will travel through that material and the higher the } } refractive index. For instance: } } RI = velocity of light in air/velocity of light in material } } RI of air = 1, velocity of light = 300,000 km/s } } RI of water is 1.33, velocity of light is 225,000 km/s } } RI of immersion oil, glass, and many polymers is approximately } } 1.5, velocity of light = 200,000 km/sec. } } } } As for refraction, the analogy which is often used is that of a } } person roller skating (or in this day and age, roller blading) from } } one surface to another. For example, from the concrete sidewalk } } onto grass. The smooth surface of the concrete is analogous to a } } material with low refractive index; the rougher surface of glass is } } analogous to a material with higher RI. If the skater approaches } } the sidewalk:grass boundary with both feet parallel, both feet will } } slow down by the same amount and the skater will continue skating } } in the same direction. However, if the skater approaches the } } boundary at an angle, one foot will slow down while the other } } remains at the original speed. That different in speed causes the } } skater to pivot toward the material of higher RI. } } } } If you think of light in terms of a wave front rather than a } } simple ray, the analogy transfers easily. If the wave approaches a } } boundary at an angle, the edge of the front which hits the higher } } RI first will slow down, causing the whole wave front to pivot } } around that point. That's what happens, for example, when light } } travels from air (RI = 1.00) into a glass coverslip (RI ~1.5) or a } } droplet of water or a cell (RI ~1.33). You can use the optical } } axis of the microscope as a reference. In these cases, the light } } will bend in TOWARD the optical axis. } } Conversely, when light emerges from a glass slide or a droplet of } } water it will bend AWAY from the OA. Actually this is why we use } } oil immersion: to cause those rays which would normally bend away } } from the OA, causing a loss of both intensity and the critical } } contribution to resolution and edge definition, to bend back toward } } the OA, where they have an opportunity to be captured by the } } objective and make a positive contribution to better imaging. } } } } All of this is explained in detail, with diagrams in Optimizing } } Light Microscopy. If you are interested in a copy, please contact } } Ken Piel here in the MME office for purchasing details (see below). } } } } 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 next } } April. 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 or email him at kenpiel-at-mme1.com } } } } } } } } At 08:24 AM 12/6/2006, vthawfeek-at-yahoo.co.in wrote: } } } } } } } } } } } } } 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 vthawfeek-at-yahoo.co.in as well as the } } MIcroscopy Listserver } } } --------------------------------------------------------------------------- } } } } } } Email: vthawfeek-at-yahoo.co.in } } } Name: V.Thawfeek Mohammed } } } } } } Organization: psg tech coll. of engg. } } } } } } Title-Subject: [Filtered] optical physics } } } } } } Question: why does a light beam diverge when it strikes a medium } } with greater refractive index than the medium ,in which it is. } } } } } } --------------------------------------------------------------------------- } } }
On Dec 6, 2006, at 12:25 PM, baskin-at-bio.umass.edu wrote:
} These and other wonderful answers to the Snell law question } have all been wave-based. Is there an equivalent photon-based answer? } Presumably, if you fired one photon at a time at an oblique surface } you would get the most counts at the Snell angle; but, with single } photons, how are we to understand lines of coherent marchers or even } two feet on roller blades? Is this light being weird? } Dear Tobias, The way to look at a single photon is that it consists of a wave packet with a distribution of wavelengths, so that it is pretty much localized and has a reasonably defined momentum, but neither is exact. Then each wave that makes up the wave packet scatters independently resulting in a set of waves distributed around the Snell angle. These waves combine to give you the single photon in the second medium--remember, waves obey the law of superposition; the amplitudes add with the appropriate phases. There is an excellent analogy that you can demonstrate in your scope. If you are examining a crystal in diffraction mode with a beam current so low that only one electron is coming down the column at any time--calculations will convince you that this is achievable with small C2 aperture and high spot size in most scopes--the electron will scatter off the crystal and land on the detector, where it will register in one pixel (approximately, since the electron can scatter again and activate nearby pixels also). Thus, a simple ED experiment has the electron being produced at the gun as a particle, scattering off the specimen as a wave, then activating the detector as a particle again, a clear example of the wave-particle duality. Unless you are very comfortable with quantum mechanics, this is, indeed, weird. 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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I have 2 "blöde" questions, not directly related with microscopy, but perhaps one of you can still answer them:
1) I stored LM cryo-sections (unfixed, 5 µm thick) at -28°C for approx. 5 months and now I test the activity of alkaline phosphatase (very positive at the moment of cutting) I see no signal. Control show very strong signal, so it does not come from the technique. I wonder how I lost the enzyme activity. I know for proteins it is better to store à -80°C but I wouldn't expect to lose all the activity! The freezer is a "frost-free" freezer. Does someone know how these freezers work? Could the mechanism of frost-free freezers be detrimental to the enzymatic activity?
2) For the cell line OE21 in the ECACC catalog, I see "Hazard: CX". No other information! I couldn't find this cell line in the ATCC catalog and I couldn't find an explanation on the internet. What does it mean, CX?
About the cleaning of the turbo pump, Pfeiffer indicate for their pumps to soak them in Freon 113 or trichloréthylène, the high-vac side first, as deep as you reach the painted part of the boddy (that mean only the stainles steel part). By that way one can rince the rotor and stator of the pump. I've done this a lot of time, and at the first soak, the solvant become more or less yellow. You change then the solvent until it stays uncolored. Than you can blow hot air through the rouging port, or close the pump with an flange, and pump on it with a roughing pump, to evaporate the residue of solvent.
I suppose one can do that with all turbo pumps, at the condion that the bearing are mounted on both side of the motor. That the case in all with non-magnetic bearing. Pfeiffer's pumps have a magnetic bearing at the high-vac side of the rotor, and a ball bearing at the other end of the motor. Of coarse, one must choose a solvent which disolve the contaminent, and not the viton gasket which protect the bearing.
I had until now much success with that way, seeing a real difference in vaccum performence, as well in total pressure as in pollution measurement with a RGA.
Of coarse, for a complet cleaning, with a heavy pollution, on must desmantle the pump. That's an other stuff ! But it's worth to try the soaking methode, before sending the pump back to the manufacturer.
Hope it's help
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
john.mardinly-at-intel.com a écrit : } ---------------------------------------------------------------------------- } 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 } ---------------------------------------------------------------------------- } } Scott; } Thanks to you and everyone that responded with their thoughts on } this problem. The feeling among our group now is that your thought that } outgassing from the grease in the mechanical part of the diaphragm pumps } can backstream when the diaphragms fail is the most likely diagnosis. } This particular pump had a huge tear in the failed diaphragm, but this } was a bit of a surprise, since our many Gatan PIPS have exactly the same } pumps, and we routinely run them until diaphragm failure, although they } tend to fail with smaller tears, and perhaps the gas load from the argon } ion guns tends to keep them cleaner by purging backstreaming. } We have cleaned the vacuum chamber with solvents (and it was a } mess), but cleaning the turbo pump will be difficult, since it cannot be } disassembled. A nitrogen purge will probably be the best method of } cleaning the turbo. We also now intend to proactively change all of our } diaphragms on an annual basis, rather than running them to failure, in } the hope that this sort of mess never recurs. } } John Mardinly } Intel Corporation } } Disclaimer: This is a communication from this author and does not } represent } an opinion of Intel Corporation. } } -----Original Message----- } X-from: Scott Walck [mailto:walck-at-southbaytech.com] } Sent: Friday, December 01, 2006 2:43 PM } To: Microscopy-at-microscopy.com } Cc: Mardinly, John } Subject: RE: [Microscopy] Contamination of Vacuum Specimen Storage } System } } John, } I think that the sulfur came from the rubber diaphragms. I'm surprised, } because I thought that they would have been VitonR. Either that or } lubrication oil from moving parts that came through the failed diaphragm } and } was exposed to the system. Was the diaphragm torn or had holes in it? } Sulfur's atomic weight just might let it get through a turbo pump. } Anyway, } I think what you might be able to do is "getter" the sulfur in your } system. } Get some copper powder and put it in your system for a while. The } smaller } the powder particles and the more open the container, the better } gettering } that you will have. Silver ought to work also. It will take some time } to } getter the sulfur in your system out, but given some time, it should be } able } to do it. You will also be able to monitor the progress on the copper } particles if you periodically change the copper. } } In the mean time, you might want to consider storing your samples with } our } SampleSaverT system. We have a specially vented TEM storage box for the } SampleSaverT container system that will take TEM grids. If the samples } are } FIB cuts, you can store them in the SampleSaverT container in our } FortressT } FIB holders that will also fit into the SampleSaverT container. The } SampleSaverT containers worked for my cross section samples of Low-E } glass } where there are two layers of Ag exposed. Unfortunately, I haven't } gotten } any feedback yet from how well they work for copper metallization } samples. } I expect them to work as well there, but as I said, I don't have hard } evidence, yet. } } BTW, Generally, the suggested maintenance replacement time on diaphragms } for } pumps is about a year. } } Disclaimer: South Bay Technology, Inc. makes and sells the SampleSaverT } container and FortressT FIB holders. } } } -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: john.mardinly-at-intel.com [mailto:john.mardinly-at-intel.com] } Sent: Friday, December 01, 2006 11:40 AM } To: Walck-at-SouthBayTech.com } Subject: [Microscopy] Contamination of Vacuum Specimen Storage System } } } } } ------------------------------------------------------------------------ } ---- } 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 a Gatan Model 655 Dry Pumping Station } (www.gatan.com/holders/655_drypumpingstation.html) equipped with pods } for } storage of TEM specimens. In particular, TEM cross-sections of } integrated } circuits using copper interconnects seem to be particularly vulnerable } to } corrosion in atmosphere. This vacuum station has been essential in } protecting the specimens that we put so much work into making, and has } been } very successful for quite some time. The dry vacuum system consists of a } 'drag' pump, which is essentially a turbo pump, backed by a diaphragm } type } rough pump. Last summer, one of the rubber diaphragms failed in the } backing } pump, and the system ran for a few days with degraded vacuum. After the } diaphragms were replaced and the high vacuum restored to the E-5 Torr } range, } we found that TEM cross-sections of integrated circuits using copper } interconnects were destroyed in just a few hours if stored in this } vacuum } system. EDX of the copper corrosion product showed the presence of } sulfur. } We have tried cleaning the storage pods, and part of the inside of the } vacuum chamber, but have been totally frustrated in being able to } restore } the essential function of this vacuum storage system. Any suggestions as } to } what may have happened, and what we can do to clean up the system would } be } very much appreciated. Thank you. } } John Mardinly } Intel Corporation } } Disclaimer: This is a communication from this author and does not } represent } an opinion of Intel Corporation. } } } } ==============================Original } Headers============================== } 6, 30 -- From john.mardinly-at-intel.com Fri Dec 1 13:35:49 2006 6, 30 -- } Received: from mga01.intel.com (mga01.intel.com [192.55.52.88]) } 6, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id } kB1JZnHb024448 } 6, 30 -- for {Microscopy-at-msa.microscopy.com} ; Fri, 1 Dec 2006 } 13:35:49 -0600 } 6, 30 -- Received: from fmsmga001.fm.intel.com ([10.253.24.23]) } 6, 30 -- by mga01.intel.com with ESMTP; 01 Dec 2006 11:35:32 -0800 } 6, 30 -- Received: from scsmsx332.sc.intel.com (HELO } scsmsx332.amr.corp.intel.com) ([10.3.90.6]) } 6, 30 -- by fmsmga001.fm.intel.com with ESMTP; 01 Dec 2006 11:35:32 } -0800 } 6, 30 -- X-ExtLoop1: 1 } 6, 30 -- X-IronPort-AV: i="4.09,486,1157353200"; } 6, 30 -- d="scan'208"; a="171762165:sNHT19309206" } 6, 30 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by } scsmsx332.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); } 6, 30 -- Fri, 1 Dec 2006 11:35:31 -0800 } 6, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 6, 30 -- } Content-class: urn:content-classes:message 6, 30 -- MIME-Version: 1.0 6, } 30 } -- Content-Type: text/plain; } 6, 30 -- charset="us-ascii" } 6, 30 -- Subject: Contamination of Vacuum Specimen Storage System 6, 30 } -- } Date: Fri, 1 Dec 2006 11:35:31 -0800 6, 30 -- Message-ID: } {1DF4C4D62339DB4C9C98DF04213995B601F0A84F-at-scsmsx413.amr.corp.intel.com} } 6, 30 -- X-MS-Has-Attach: } 6, 30 -- X-MS-TNEF-Correlator: } 6, 30 -- Thread-Topic: Contamination of Vacuum Specimen Storage System } 6, } 30 -- Thread-Index: } AccVcgLpZR8yy6qqRpi55yhxXPXijQAAIjDgAAAaArAAAVCSIAAB3rtQ } 6, 30 -- From: "Mardinly, John" {john.mardinly-at-intel.com} 6, 30 -- To: } {Microscopy-at-msa.microscopy.com} 6, 30 -- X-OriginalArrivalTime: 01 Dec } 2006 } 19:35:31.0926 (UTC) FILETIME=[DD2F8F60:01C7157F] 6, 30 -- } Content-Transfer-Encoding: 8bit 6, 30 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id kB1JZnHb024448 } ==============================End of - } Headers============================== } } } ==============================Original Headers============================== } 26, 30 -- From john.mardinly-at-intel.com Wed Dec 6 10:56:04 2006 } 26, 30 -- Received: from mga02.intel.com (mga02.intel.com [134.134.136.20]) } 26, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB6Gu3dq024503 } 26, 30 -- for {Microscopy-at-microscopy.com} ; Wed, 6 Dec 2006 10:56:03 -0600 } 26, 30 -- Received: from orsmga001.jf.intel.com ([10.7.209.18]) } 26, 30 -- by mga02.intel.com with ESMTP; 06 Dec 2006 08:51:57 -0800 } 26, 30 -- Received: from scsmsx331.sc.intel.com (HELO scsmsx331.amr.corp.intel.com) ([10.3.90.4]) } 26, 30 -- by orsmga001.jf.intel.com with ESMTP; 06 Dec 2006 08:51:55 -0800 } 26, 30 -- X-ExtLoop1: 1 } 26, 30 -- X-IronPort-AV: i="4.09,505,1157353200"; } 26, 30 -- d="scan'208"; a="171184232:sNHT45583097" } 26, 30 -- Received: from scsmsx413.amr.corp.intel.com ([10.3.90.32]) by scsmsx331.amr.corp.intel.com with Microsoft SMTPSVC(6.0.3790.1830); } 26, 30 -- Wed, 6 Dec 2006 08:51:52 -0800 } 26, 30 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 26, 30 -- Content-class: urn:content-classes:message } 26, 30 -- MIME-Version: 1.0 } 26, 30 -- Content-Type: text/plain; } 26, 30 -- charset="us-ascii" } 26, 30 -- Subject: RE: [Microscopy] Contamination of Vacuum Specimen Storage System } 26, 30 -- Date: Wed, 6 Dec 2006 08:51:52 -0800 } 26, 30 -- Message-ID: {1DF4C4D62339DB4C9C98DF04213995B601FBA503-at-scsmsx413.amr.corp.intel.com} } 26, 30 -- X-MS-Has-Attach: } 26, 30 -- X-MS-TNEF-Correlator: } 26, 30 -- Thread-Topic: [Microscopy] Contamination of Vacuum Specimen Storage System } 26, 30 -- Thread-Index: AccVgIJVL6E6+CDvTlWJF2VEgMf9YAAE2UawAPA2/rA= } 26, 30 -- From: "Mardinly, John" {john.mardinly-at-intel.com} } 26, 30 -- To: "Scott Walck" {walck-at-southbaytech.com} , {Microscopy-at-microscopy.com} } 26, 30 -- X-OriginalArrivalTime: 06 Dec 2006 16:51:52.0884 (UTC) FILETIME=[D4A56740:01C71956] } 26, 30 -- Content-Transfer-Encoding: 8bit } 26, 30 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kB6Gu3dq024503 } ==============================End of - Headers============================== }
==============================Original Headers============================== 12, 30 -- From jacques.faerber-at-ipcms.u-strasbg.fr Thu Dec 7 02:39:36 2006 12, 30 -- Received: from mailhost.u-strasbg.fr (mailhost.u-strasbg.fr [130.79.200.157]) 12, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB78dZ4e031374 12, 30 -- for {microscopy-at-microscopy.com} ; Thu, 7 Dec 2006 02:39:36 -0600 12, 30 -- Received: from ipcms.u-strasbg.fr (ipcms.u-strasbg.fr [130.79.210.2]) 12, 30 -- by mailhost.u-strasbg.fr (8.13.8/jtpda-5.5pre1) with ESMTP id kB78dWTS099809 12, 30 -- for {microscopy-at-microscopy.com} ; Thu, 7 Dec 2006 09:39:32 +0100 (CET) 12, 30 -- Received: from [130.79.54.3] (odhinn.u-strasbg.fr [130.79.54.3]) 12, 30 -- by ipcms.u-strasbg.fr (Postfix) with ESMTP id 7F4873EC114 12, 30 -- for {Microscopy-at-Microscopy.Com} ; Thu, 7 Dec 2006 09:38:48 +0100 (CET) 12, 30 -- Message-ID: {4577D32D.9040408-at-ipcms.u-strasbg.fr} 12, 30 -- Date: Thu, 07 Dec 2006 09:39:09 +0100 12, 30 -- From: "j.faerber" {jacques.faerber-at-ipcms.u-strasbg.fr} 12, 30 -- User-Agent: Thunderbird 1.5.0.8 (X11/20061117) 12, 30 -- MIME-Version: 1.0 12, 30 -- To: Microscopy-at-microscopy.com 12, 30 -- Subject: Re: [Microscopy] RE: Contamination of Vacuum Specimen Storage System 12, 30 -- + turbo pump 12, 30 -- References: {200612061703.kB6H3tTP011952-at-ns.microscopy.com} 12, 30 -- In-Reply-To: {200612061703.kB6H3tTP011952-at-ns.microscopy.com} 12, 30 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 12, 30 -- Content-Transfer-Encoding: 8bit 12, 30 -- X-IPCMS-MailScanner: Found to be clean 12, 30 -- X-IPCMS-MailScanner-From: jacques.faerber-at-ipcms.u-strasbg.fr 12, 30 -- X-Greylist: Sender IP whitelisted, not delayed by milter-greylist-2.0.2 (mailhost.u-strasbg.fr [130.79.200.157]); Thu, 07 Dec 2006 09:39:32 +0100 (CET) 12, 30 -- X-Virus-Scanned: ClamAV 0.88.5/2299/Thu Dec 7 08:36:50 2006 on mr7.u-strasbg.fr 12, 30 -- X-Virus-Status: Clean 12, 30 -- X-Spam-Status: No, score=-0.5 required=5.0 tests=AWL autolearn=disabled 12, 30 -- version=3.1.7 12, 30 -- X-Spam-Checker-Version: SpamAssassin 3.1.7 (2006-10-05) on mr7.u-strasbg.fr ==============================End of - Headers==============================
actually Newton came up with a particle-theory of refraction.
The photons are accelerated (Don't ask me how) at the surface from lower refractive index to higher. This acceleration is normal to the surface and leaves velocity-components parallel to the surface unaffected. Thus the velocity-vector changes direction towards the normal.
Notice that this theory requires higher velocity in the high-index medium, whereas the wave-expalnation requires lower velocity. The answer to this puzzle is that the wave-explanation uses the phase-velocity whereas, speaking in wave-terms, Newton would have been talking about the group-velocity, which Bill Tivol refers to in his mail. In fact in a dispersive medium (different colours have different phase-velocities) the group- and phase-velocities are different. The group-velocity can actually be larger, in support of Newton.
Philip
-----Original Message----- X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu] Sent: 06 December 2006 21:28 To: Philip.Koeck-at-biosci.ki.se
Mohammed,
some clarification would be helpful. Do you really mean "diverge" or do you mean "split into different colors"?
An already divergent monochromatic beam should actually diverge less after passing into the higher refractive index.
Philip
-----Original Message----- X-from: dljones-at-bestweb.net [mailto:dljones-at-bestweb.net] Sent: 06 December 2006 19:04 To: Philip.Koeck-at-biosci.ki.se
This thread is following the route to a Nobel prize in Physics.. have a read of Richard Feynman's "QED-the strange theory of light and matter", which addresses exactly this point and is a summary of the work for which he was awarded his gong. I haven't read it for a few years, but I remember him starting with glass being 80% transmissive, and asking how could individual photons 'know' whether to cross the glass or not to give the right answer of 80%. Like most of his stuff it is very readable and entertaining. And yes, the world is actually weirder than you think it is..
-----Original Message----- X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu] Sent: 06 December 2006 20:27 To: Richard Beanland
Greetings, These and other wonderful answers to the Snell law question have all been wave-based. Is there an equivalent photon-based answer? Presumably, if you fired one photon at a time at an oblique surface you would get the most counts at the Snell angle; but, with single photons, how are we to understand lines of coherent marchers or even two feet on roller blades? Is this light being weird?
Just wondering... Tobias
} This is a really fine answer. I would add one more analogy, since } the question (I think) relates to the change in direction of the path } of the light beam. In my classes, I refer to the image of a column } of a marching band going from a concrete surface to a broken field. } If they are exactly perpendicular to the boundary, they continue to } go straight, although with some stumbles. If they enter at an angle, } then those members who are on the "acute" side of the column slow } down first. This "pulls the others along as well, bit by bit, and } the overall direction of the march changes. } } Date sent: Wed, 6 Dec 2006 11:21:25 -0600 } To: jbs-at-temple.edu } X-from: bfoster-at-mme1.com } Send reply to: bfoster-at-mme1.com } Subject: [Microscopy] Re: viaWWW: optical physics } } } } } } } } } } } } Hi, } } } } The answer is really quite simple: Snell's Law. The bending of } } light as it crosses the boundary from one RI to another is governed } } by the Laws of Refraction. } } } } The bending discussed in your question requires that several } } conditions be met: First, that there be different RIs on each side } } of the boundary. Second, that the light approach the boundary at } } an angle. } } } } To understand what happens, it is first necessary to understand } } that refractive index is actually a measure of the impact of } } interaction between the electric field of light and the electric } } field of matter. The greater the interaction, the more slowly } } light will travel through that material and the higher the } } refractive index. For instance: } } RI = velocity of light in air/velocity of light in material } } RI of air = 1, velocity of light = 300,000 km/s } } RI of water is 1.33, velocity of light is 225,000 km/s } } RI of immersion oil, glass, and many polymers is approximately } } 1.5, velocity of light = 200,000 km/sec. } } } } As for refraction, the analogy which is often used is that of a } } person roller skating (or in this day and age, roller blading) from } } one surface to another. For example, from the concrete sidewalk } } onto grass. The smooth surface of the concrete is analogous to a } } material with low refractive index; the rougher surface of glass is } } analogous to a material with higher RI. If the skater approaches } } the sidewalk:grass boundary with both feet parallel, both feet will } } slow down by the same amount and the skater will continue skating } } in the same direction. However, if the skater approaches the } } boundary at an angle, one foot will slow down while the other } } remains at the original speed. That different in speed causes the } } skater to pivot toward the material of higher RI. } } } } If you think of light in terms of a wave front rather than a } } simple ray, the analogy transfers easily. If the wave approaches a } } boundary at an angle, the edge of the front which hits the higher } } RI first will slow down, causing the whole wave front to pivot } } around that point. That's what happens, for example, when light } } travels from air (RI = 1.00) into a glass coverslip (RI ~1.5) or a } } droplet of water or a cell (RI ~1.33). You can use the optical } } axis of the microscope as a reference. In these cases, the light } } will bend in TOWARD the optical axis. } } Conversely, when light emerges from a glass slide or a droplet of } } water it will bend AWAY from the OA. Actually this is why we use } } oil immersion: to cause those rays which would normally bend away } } from the OA, causing a loss of both intensity and the critical } } contribution to resolution and edge definition, to bend back toward } } the OA, where they have an opportunity to be captured by the } } objective and make a positive contribution to better imaging. } } } } All of this is explained in detail, with diagrams in Optimizing } } Light Microscopy. If you are interested in a copy, please contact } } Ken Piel here in the MME office for purchasing details (see below). } } } } 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 next } } April. 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 or email him at kenpiel-at-mme1.com } } } } } } } } At 08:24 AM 12/6/2006, vthawfeek-at-yahoo.co.in wrote: } } } } } } } } } } } } } 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 vthawfeek-at-yahoo.co.in as well as the } } MIcroscopy Listserver } } } ----------------------------------------------------------------------- ---- } } } } } } Email: vthawfeek-at-yahoo.co.in } } } Name: V.Thawfeek Mohammed } } } } } } Organization: psg tech coll. of engg. } } } } } } Title-Subject: [Filtered] optical physics } } } } } } Question: why does a light beam diverge when it strikes a medium } } with greater refractive index than the medium ,in which it is. } } } } } } ----------------------------------------------------------------------- ---- } } }
==============================Original Headers============================== 6, 19 -- From baskin-at-bio.umass.edu Wed Dec 6 14:25:31 2006 6, 19 -- Received: from marlin.bio.umass.edu (marlin.bio.umass.edu [128.119.55.19]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB6KPVfD011840 6, 19 -- for {microscopy-at-microscopy.com} ; Wed, 6 Dec 2006 14:25:31 -0600 6, 19 -- Received: from [172.30.55.79] (eutopia [128.119.55.30]) 6, 19 -- by marlin.bio.umass.edu (8.13.6/8.13.6) with ESMTP id kB6KPQZB025363; 6, 19 -- Wed, 6 Dec 2006 15:25:27 -0500 (EST) 6, 19 -- Mime-Version: 1.0 6, 19 -- Message-Id: {p0623090dc19cd66dc686-at-[172.30.55.79]} 6, 19 -- In-Reply-To: {200612062000.kB6K0oST001676-at-ns.microscopy.com} 6, 19 -- References: {200612062000.kB6K0oST001676-at-ns.microscopy.com} 6, 19 -- Date: Wed, 6 Dec 2006 15:25:24 -0500 6, 19 -- To: microscopy-at-microscopy.com 6, 19 -- From: Tobias Baskin {baskin-at-bio.umass.edu} 6, 19 -- Subject: optical physics:Challenge 6, 19 -- Cc: jbs-at-temple.edu 6, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 19 -- X-Greylist: Sender succeded SMTP AUTH authentication, not delayed by milter-greylist-1.6 (marlin.bio.umass.edu [128.119.55.19]); Wed, 06 Dec 2006 15:25:29 -0500 (EST) 6, 19 -- X-Scanned-By: MIMEDefang 2.54 on 128.119.55.19 ==============================End of - Headers==============================
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==============================Original Headers============================== 15, 32 -- From richard.beanland-at-bookham.com Thu Dec 7 03:45:10 2006 15, 32 -- Received: from mail83.messagelabs.com (mail83.messagelabs.com [195.245.231.83]) 15, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kB79j90N032413 15, 32 -- for {microscopy-at-microscopy.com} ; Thu, 7 Dec 2006 03:45:10 -0600 15, 32 -- X-VirusChecked: Checked 15, 32 -- X-Env-Sender: richard.beanland-at-bookham.com 15, 32 -- X-Msg-Ref: server-15.tower-83.messagelabs.com!1165484708!42046564!1 15, 32 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 15, 32 -- X-Originating-IP: [213.249.209.179] 15, 32 -- Received: (qmail 31146 invoked from network); 7 Dec 2006 09:45:08 -0000 15, 32 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 15, 32 -- by server-15.tower-83.messagelabs.com with SMTP; 7 Dec 2006 09:45:08 -0000 15, 32 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 15, 32 -- Thu, 7 Dec 2006 09:45:15 +0000 15, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 15, 32 -- Content-class: urn:content-classes:message 15, 32 -- MIME-Version: 1.0 15, 32 -- Content-Type: text/plain; 15, 32 -- charset="us-ascii" 15, 32 -- Subject: RE: [Microscopy] optical physics:Challenge 15, 32 -- Date: Thu, 7 Dec 2006 09:45:14 -0000 15, 32 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E2CC54D-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 15, 32 -- X-MS-Has-Attach: 15, 32 -- X-MS-TNEF-Correlator: 15, 32 -- Thread-Topic: [Microscopy] optical physics:Challenge 15, 32 -- thread-index: AccZdN5xFl4Td6sqTl6++eu/sEqL/AAbkU2w 15, 32 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 15, 32 -- To: {baskin-at-bio.umass.edu} 15, 32 -- Cc: {microscopy-at-microscopy.com} 15, 32 -- X-OriginalArrivalTime: 07 Dec 2006 09:45:15.0631 (UTC) FILETIME=[65E84FF0:01C719E4] 15, 32 -- Content-Transfer-Encoding: 8bit 15, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kB79j90N032413 ==============================End of - Headers==============================
Hello Stephane, The frost-free freezer may have been the culprit. They keep and average temp of -20, but the temp. cycles so that the frost can be cleared. I have one in my lab, too, but I keep anything that may be really temp. sensitive in a separate -20 that is definitely NOT frost free (we just spent 2 days defrosting it!). On the other hand, I have also kept cryo sections of chicken heart in the frost-free -20 for months and continued to have good immuno labelling. I think perhaps that the enzymes are more sensitve to freeze-thaw. 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 Thu Dec 7 08:42:56 2006 1, 24 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 1, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB7Egufc018796 1, 24 -- for {microscopy-at-microscopy.com} ; Thu, 7 Dec 2006 08:42:56 -0600 1, 24 -- Received: from mpx3.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 1, 24 -- by smtp-gw1.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id kB7EgdjE011089; 1, 24 -- Thu, 7 Dec 2006 09:42:53 -0500 (EST) 1, 24 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 1, 24 -- by mpx3.med.cornell.edu 1, 24 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 1, 24 -- with ESMTPA id {0J9W00GKVRIY1610-at-mpx3.med.cornell.edu} ; Thu, 1, 24 -- 07 Dec 2006 09:42:35 -0500 (EST) 1, 24 -- Date: Thu, 07 Dec 2006 09:42:32 -0500 1, 24 -- From: "Leona Cohen-Gould" {lcgould-at-med.cornell.edu} 1, 24 -- Subject: Re: [Microscopy] 2 questions not directly related to microscopy 1, 24 -- In-reply-to: {200612070754.kB77sR43020952-at-ns.microscopy.com} 1, 24 -- Sender: "Leona Cohen-Gould" {lcgould-at-med.cornell.edu} 1, 24 -- To: nizets2-at-yahoo.com, Microscopy Listserver {microscopy-at-microscopy.com} 1, 24 -- Message-id: {p06230905c19dd7efca5e-at-[140.251.48.23]} 1, 24 -- MIME-version: 1.0 1, 24 -- Content-type: text/plain; charset=us-ascii; format=flowed 1, 24 -- Content-transfer-encoding: 7BIT 1, 24 -- References: {200612070754.kB77sR43020952-at-ns.microscopy.com} 1, 24 -- X-PMX-Version: 5.2.2.285561, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2006.12.7.62432 ==============================End of - Headers==============================
Rhonda, We have had our Leica EM Stain unit for about 2 years now. I love it and would recommend it's purchase. Just be aware of a number of things:
1. Leica can only ship bottled 0.5% uranyl stain. I feel my epoxy resins are too weakly stained with this so I modify to 2%. 2. I prefer the 40-grid Hiroaka Plates from Electron Microscopy Sciences rather than the Leica 25-grid plates. The Hiroaka plates seem to be made of a different plastic and maintain their flexibility longer (I discard after about 10 runs). Unfortunately I found the Leica plates start to loose their grip on the grids after a couple of uses. 3. I have set the system to use 15ml of stain per run but you have the option of 5-25ml in 5ml increments. 3. Adherence to a rigorous system cleaning schedule is extremely important for section cleanliness. The system should be stored in an "Extended Wash" cycle using 3% nitric acid. Prior to staining run, boil and cool ~2L dH2O. Finish the extended wash cycle, run and finish a new extended wash cycle, and only THEN run your wash cycle. This can take 1.5 hrs. Now you can stain (~1 hr procedure).
The times may sound long, but you simply walk away and do something else. You can even set it up with a delay function to start early the next morning and finish when just as you're walking in. If you want my system details, give a holler.
Best regards,
Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"The ultimate human freedom is the ability to choose one's attitude in a given set of circumstances." -Viktor Frankl
-----Original Message----- X-from: rra-at-stowers-institute.org [mailto:rra-at-stowers-institute.org] Sent: Tuesday, December 05, 2006 5:33 PM To: Bobrowski, Walter
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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
==============================Original Headers============================== 7, 12 -- From zaluzec-at-microscopy.com Tue Dec 5 16:26:17 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 kB5MQGaV013340 7, 12 -- for {microscopy-at-microscopy.com} ; Tue, 5 Dec 2006 16:26:17 -0600 7, 12 -- Mime-Version: 1.0 7, 12 -- X-Sender: (Unverified) 7, 12 -- Message-Id: {p06110400c19ba26526ef-at-[206.69.208.22]} 7, 12 -- Date: Tue, 5 Dec 2006 16:26:16 -0600 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============================== ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately.
==============================Original Headers============================== 22, 29 -- From Walter.Bobrowski-at-pfizer.com Thu Dec 7 08:51:14 2006 22, 29 -- Received: from gromsgom01.pfizer.com (gromsgo.pfizer.com [148.168.224.84]) 22, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB7EpE6t001284 22, 29 -- for {Microscopy-at-microscopy.com} ; Thu, 7 Dec 2006 08:51:14 -0600 22, 29 -- Received: from mopamrexc02.amer.pfizer.com (mopamrexc02.pfizer.com [170.116.30.68]) 22, 29 -- by gromsgom01.pfizer.com (8.13.4/8.13.4) with ESMTP id kB7Eol31014318; 22, 29 -- Thu, 7 Dec 2006 09:50:47 -0500 22, 29 -- Received: from mopamrexc01.amer.pfizer.com ([170.116.32.254]) by mopamrexc02.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 22, 29 -- Thu, 7 Dec 2006 09:51:13 -0500 22, 29 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 22, 29 -- Thu, 7 Dec 2006 09:51:13 -0500 22, 29 -- X-MIMEOLE: Produced By Microsoft Exchange V6.5.7226.0 22, 29 -- Content-class: urn:content-classes:message 22, 29 -- MIME-Version: 1.0 22, 29 -- Content-Type: text/plain; 22, 29 -- charset="US-ASCII" 22, 29 -- Subject: RE: [Microscopy] viaWWW: Leica EM-Stain 22, 29 -- Date: Thu, 7 Dec 2006 09:51:11 -0500 22, 29 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D082A1338-at-anaamrexm01.amer.pfizer.com} 22, 29 -- X-MS-Has-Attach: 22, 29 -- X-MS-TNEF-Correlator: 22, 29 -- Thread-Topic: [Microscopy] viaWWW: Leica EM-Stain 22, 29 -- Thread-Index: AccYvbSnWhtxHadlRT6pxmbEKRP8QgBReZmw 22, 29 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 22, 29 -- To: {rra-at-stowers-institute.org} , {Microscopy-at-microscopy.com} 22, 29 -- X-OriginalArrivalTime: 07 Dec 2006 14:51:13.0295 (UTC) FILETIME=[23ED89F0:01C71A0F] 22, 29 -- X-Proofpoint-Spam-Reason: safe 22, 29 -- Content-Transfer-Encoding: 8bit 22, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kB7EpE6t001284 ==============================End of - Headers==============================
Owen, Seal the BEEM cap with some nail polish (or whatever) and turn upside down. This will let the long axis of the wood be parallel to the surface. If you want endgrain endon, after polymerization, use a saw of some kind and cut a few mm "slice" off the end and then trim and mount upright on a blank block. Superglue bonds to epoxies really tightly so this works, though admittedly inelegant. You can also buy BEEM style caps (they are in fact TAAB capsules) that have flat bottoms so you don't have to seal the tops and flip.
Hope this helps, Tobias
} } All, } } How would one go about embedding wood? I am having trouble orienting } the end grain in a BEEM capsule. } } Owen Mills } Michigan Tech University }
7-Dec-2006 The Dow Chemical Company has an opening for a Ph.D. Chemist, Polymer Scientist or Materials Scientist in the microscopy discipline of our Global Analytical Sciences Laboratory.
Location of current position: Schkopau, Germany Job Title: Senior Research Chemist Degree: Ph.D. Discipline: Analytical Chemistry, Physical Chemistry, Materials Science, Physics, or Polymer Science
Job Description Overview: An immediate opening exists for a skilled microscopist in Dow's Global Analytical Sciences Laboratory within 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. Candidate will be responsible for using microscopy and microanalysis to solve a wide range of materials problems in multiple application areas. The candidate will be expected to work as part of a cross-functional team in partnership with Dow R&D personnel in a variety of technology areas including catalysis, polymer science, and materials science.
For this position we are seeking candidates who are proficient in a variety of microscopy techniques. Working experience in electron microscopy and/or scanned probe microscopy is essential. Additional skills in ultramicrotomy, energy dispersive x-ray analysis, and capability in other microscopy techniques and surface science techniques such as optical, TEM, and XPS is preferred. Candidates must have excellent problem solving skills, a background in polymer science is a plus. Ph.D. in Analytical Chemistry, Physical Chemistry, Physics, Engineering or Materials Science is preferred, but an individual with a Masters degree and relevant work experience will also be considered. The position is located in Schkopau, Germany (Halle/Leipzig area). Fluency in English is required, fluency in German is a plus.
If you are highly qualified and are interested in this job, please submit your electronic resume and curriculum vitae directly to tdominowski-at-dow.com or send paper copy to: Dr. Georg Bar Dow Olefinverbund GmbH PF 1163, 06201 Merseburg, Germany.
Best Regards,
Bill William A. Heeschen, Ph.D. Microscopy, Digital Imaging The Dow Chemical Company Midland, MI 48674 waheeschen-at-dow.com
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.
==============================Original Headers============================== 12, 23 -- From WAHeeschen-at-dow.com Thu Dec 7 09:23:48 2006 12, 23 -- Received: from mail130.messagelabs.com (mail130.messagelabs.com [216.82.250.163]) 12, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kB7FNmEG030204 12, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 7 Dec 2006 09:23:48 -0600 12, 23 -- X-VirusChecked: Checked 12, 23 -- X-Env-Sender: WAHeeschen-at-dow.com 12, 23 -- X-Msg-Ref: server-15.tower-130.messagelabs.com!1165505026!4106543!1 12, 23 -- X-StarScan-Version: 5.5.10.7; banners=-,-,- 12, 23 -- X-Originating-IP: [216.99.65.28] 12, 23 -- Received: (qmail 4543 invoked from network); 7 Dec 2006 15:23:47 -0000 12, 23 -- Received: from mail8.dow.com (HELO mante89.nam.dow.com) (216.99.65.28) 12, 23 -- by server-15.tower-130.messagelabs.com with SMTP; 7 Dec 2006 15:23:47 -0000 12, 23 -- Received: by mante89.nam.dow.com with Internet Mail Service (5.5.2658.3) 12, 23 -- id {W5R91SGC} ; Thu, 7 Dec 2006 10:23:46 -0500 12, 23 -- Message-ID: {0F0E6B7B4F6AE84CBC5E7AF57057FBA8084EB2-at-USMDLMDOWX022.dow.com} 12, 23 -- From: "Heeschen, Bill (WA)" {WAHeeschen-at-dow.com} 12, 23 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 12, 23 -- Subject: Position available: The Dow Chemical Company - Analytical Scienc 12, 23 -- es Career Opportunities 12, 23 -- Date: Thu, 7 Dec 2006 10:23:01 -0500 12, 23 -- MIME-Version: 1.0 12, 23 -- X-Mailer: Internet Mail Service (5.5.2658.3) 12, 23 -- Content-Type: text/plain ==============================End of - Headers==============================
Hello Everyone, I find that I need to start shopping for a cryo ultra microtome. Any advise the members of this list would care to share is welcome as would be product literature and price quotes from vendors. We are targeting 2008, but we need to start floating numbers and requirements in front of upper management now.
Please feel free (gasp!) to snail or E-mail me literature.
Thanks!!!
Frank Karl
Degussa Corporation
Akron Technical Center
3500 Embassy Parkway
Suite 100
Akron, Ohio 44333 330-668-2235 Ext. 238 This e-mail transmission, and any documents, files or previous e-mail messages attached to it may contain information that is confidential or legally privileged. If you are not the intended recipient, or a person responsible for delivering it to the intended recipient, you are hereby notified that you must not read this transmission and that any disclosure, copying, printing, distribution or use of any of the information contained in or attached to this transmission is STRICTLY PROHIBITED. If you have received this transmission in error, please immediately notify the sender by telephone or return e-mail and delete the original transmission and its attachments without reading or saving in any manner. Thank you.
==============================Original Headers============================== 19, 17 -- From frank.karl-at-degussa.com Thu Dec 7 14:19:11 2006 19, 17 -- Received: from framailout1.rz.itson.com (mailout2.degussa.com [149.216.91.173]) 19, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB7KJA0U017984 19, 17 -- for {microscopy-at-msa.microscopy.com} ; Thu, 7 Dec 2006 14:19:10 -0600 19, 17 -- Received: from mobuscomm01.mail.degussa.com (uss1026.applications.degussanet.com [10.88.88.98]) 19, 17 -- by framailout1.rz.itson.com (8.13.4/8.13.4/Debian-3sarge3) with SMTP id kB7KJ7dV028928 19, 17 -- for {microscopy-at-msa.microscopy.com} ; Thu, 7 Dec 2006 21:19:08 +0100 19, 17 -- Subject: request for product (microtomes) information and prices 19, 17 -- To: microscopy-at-msa.microscopy.com 19, 17 -- X-Mailer: Lotus Notes Release 6.5.5 November 30, 2005 19, 17 -- Message-ID: {OF97E936E0.78C31E6A-ON8625723D.006EDC57-8525723D.006F9BC1-at-degussa.com} 19, 17 -- From: frank.karl-at-degussa.com 19, 17 -- Date: Thu, 7 Dec 2006 15:19:04 -0500 19, 17 -- X-MIMETrack: Serialize by Router on MOBUSComm01/DHexternal/US(Release 6.5.1|January 21, 2004) at 19, 17 -- 12/07/2006 02:19:09 PM 19, 17 -- MIME-Version: 1.0 19, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Post Doctoral Position in the NCSU Analytical Instrumentation Facility SIMS Laboratory
The Analytical Instrumentation Facility (AIF) of the NC State University College of Engineering has an opening for a postdoctoral research associate in its SIMS laboratory. The successful candidate will operate our magnetic sector IMS-6f magnetic sector and our PHI TRIFT I SIMS instruments. The successful candidate must be interested in learning and participating in the analysis of a wide variety of samples (semiconductors, metals, ceramics, polymers). Work in the laboratory will consist of a combination of SIMS analytical technique research, experimental design, instrument operation and data interpretation for university and industrial researchers. Training in SIMS and other analytical techniques and instrumentation (AFM, FIB, SEM, XPS, optical and stylus profilometry etc.) will be provided as needed for supporting SIMS research and analysis. For information on AIF, see our web site: http://www.ncsu.edu/aif/. A Ph.D in Analytical Chemistry, Applied Physics, Materials Science or a materials related discipline along with hands on experience with SIMS or other surface analysis or ion beam instrumentation is required. SIMS experience is highly desirable although training will be given to a suitable applicant. Experience in UHV equipment and/or electronics maintenance; operational and programming experience with Windows and Unix operating systems; and experience with analysis of semiconductor or other non biological materials are desirable.
Please apply on line at http://jobs.ncsu.edu ( please search position # 04-31-0601). Applicants will need to submit a cover letter, curriculum vitae and the names and addresses of three references. Review of applications will begin 1/08/2007, and this position will remain open until a suitable candidate is identified. NC State University is an OEO/AA employer. NC State welcomes all persons without regard to sexual orientation. ADA individuals desiring reasonable accommodations call 919-515-3148.
==============================Original Headers============================== 4, 19 -- From dale_batchelor-at-ncsu.edu Thu Dec 7 15:34:50 2006 4, 19 -- Received: from uni07mr.unity.ncsu.edu (uni07mr.unity.ncsu.edu [152.1.224.166]) 4, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB7LYoFf008807 4, 19 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 7 Dec 2006 15:34:50 -0600 4, 19 -- Received: from [152.14.52.68] (dbmacx.aif.ncsu.edu [152.14.52.68]) 4, 19 -- by uni07mr.unity.ncsu.edu (8.13.7/8.13.8/Nv5.2006.1109) with ESMTP id kB7LYmWI019058 4, 19 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 7 Dec 2006 16:34:49 -0500 (EST) 4, 19 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 19 -- Content-Transfer-Encoding: 7bit 4, 19 -- Message-Id: {2d570ae9e86b73e2cd5e3cbe7707ae11-at-ncsu.edu} 4, 19 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 19 -- To: Microscopy-at-msa.microscopy.com 4, 19 -- From: Dale Batchelor {dale_batchelor-at-ncsu.edu} 4, 19 -- Subject: [Microscopy] Post Doc Position 4, 19 -- Date: Thu, 7 Dec 2006 16:35:24 -0500 4, 19 -- X-Mailer: Apple Mail (2.624) 4, 19 -- X-PMX-Version: 5.2.1.279297, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2006.12.7.131933 4, 19 -- X-Spam-Status: No, Hits=7% 4, 19 -- X-Spam-Level: IIIIIII ==============================End of - Headers==============================
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Email: burrmich-at-msu.edu Name: Mike
Title-Subject: [Filtered] Sample prep for gels
Question: Can someone help me and suggest some good techniques for SEM/EDS analysis of gels. More specifically, i have a silicone based gel that i would like to run an EDS scan on without contaminating the chamber.
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Email: coridan-at-uiuc.edu Name: Robert Coridan
Organization: UIUC, IL, USA
Title-Subject: [Filtered] Vitrobot problems -- Base cracking?
Question: Hi---
We recently purchased an FEI Vitrobot and have had some success using it, but there are 2 problems that make working with it difficult.
1) The plungerod sticks: On ths model, there is a horizontal (left-to-right) adjustment to allow the operator to adjust the guidance of the plungerod. However, it seems that some front-and-back adjustments are required too. When we drop the plungerod, it is sometimes slowed (or stopped middrop) by the sliding washer because the rod is too far forward.
2) The foam base has cracked irreparably twice in the 2 months we have been using it. This is a newer design for the vitrobot base: the foam holds the LN2 and also insulates the brass ethane cup. We are using it according to the directions, but this foam is breaking more frequently than it should.
I have contacted FEI, but I also thought I'd check to see if any other Vitrobot users have experienced any of these problems.
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Email: john.grace-at-abbott.com Name: John Grace
Organization: Abbott Laboratories
Title-Subject: [Filtered] Frame Grabber boards for PCI Express Computers
Question: All,
I am replacing my aging G4 Image processing workstation. I have a Scion CG-7 framegrabber board, this will not work in the newer Mac workstations with PCI Express slots. Are there any framegrabbers that will work with Image j and a newer Mac available? Should I just get a PC with PCI slots?
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Email: mahonys-at-agr.gc.ca Name: Shannon Mahony
Organization: Agriculture and Agri-Food Canada
Title-Subject: [Filtered] Imaging of structures in glycerin
Question: Hello all,
I have been attempting to take pictures (with the microscope) of insect genitalia in glycerin. I have seen excellent, sharp results before using this method. However, I am finding that even using a Z-stack and montaging the images with AutoMontage, I am not getting a clear, sharp result. I am confident that this is not due to vibration during the imaging. Does anyone else have any suggestions? Do I need to make specific adjustments because of the glycerin (i.e. aperture) or anything like that?
Mohammed's question ramifies much further than it seems at first sight. Most physics students tend to be dropped into the quantum quagmire without being shown how it relates to earlier stuff. I will give a brief history here:
In the classical world, Hero of Alexandria found that light, traveling from one point to another by a reflection from a plane mirror, always takes the shortest possible path.
What Hero did for reflection, Fermat did for refraction. He took Huygens’s view that light should travel more slowly in a denser medium, and reformulates the principle by postulating that the light travels in a path that takes the least time! In hindsight, if c is constant then Hero and Fermat are in complete agreement. Based on his reasoning, he is able to deduce both the law of reflection and Snell’s law (n1sinQ1 = n2sinQ2).
Unfortunately, this was not accepted. The continentals believed that light travelled faster in a denser medium. This is the same as Newton’s corpuscular theory of light, but that came later. Rather, it was accepting the philosophical conclusions of Descartes.
The PRINCIPLE OF LEAST ACTION. Maupertuis developed the principle in first in optics, to square the calculations of Fermat with the generally held belief that light travels faster in glass than air. He enunciated “In all changes that take place in the universe, the sum of products of the speed of each body and the distance it moves is the least possible.”
This worked, so he extended it to mechanics. [[Note for future reference that energy × time is the units of Planck’s constant.]] He didn’t give it a very solid math background, though, but what did for him was giving a theological rider “Action is minimized through the Wisdom of God”. This led Voltaire to satirize him, driving him out of Paris, and he died on the way to Berlin.
Euler picked up the principle and gave it sound mathematics, but being the generous man that he was, he ascribed the principle itself to Maupertuis. In 1755 the young Lagrange wrote to Euler with a much slicker (in the best sense) mathematical formulation, and Euler dropped his own formulation and took up Lagrange's. From this we get LAGRANGIAN mechanics.
In the early 1800's Thomas Young demonstrated the wave nature of light, and the corpuscular theory was swept under the carpet.
Around 1830 William Rowan Hamilton was developing a more advanced treatment of optics based on the wave formulation. He extended this to mechanics, and in 1834-5 he published two papers on which it is possible to base all of mechanics and most of classical physics. In 1857 the English mathematician Arthur Cayley gave this formulation the name HAMILTONIAN. It is the ancestor of the Hamiltonian in quantum mechanics.
Louis de Broglie is famous among physicists for giving us the first wave theory of the electron in the doctoral thesis in 1924. In his memoirs he states “ The theory of light was suffering from a strange illness, which took the form of an antagonistic dualism between the waves of Fresnel and Maxwell, one the one hand, and photons on the other”. … “Thus appeared the idea of extending to matter the corpuscles-waves duality which was essental to light”. ***** “The equation which expresses that the motion of an electron is quantised has … the following significance - that Maupertuis’ integral [sign]mvds taken along the closed trajectory is equal to an integral multiple of [Planck’s] constant h.”
----------------------------------- Robert H. Olley URL: http://www.rdg.ac.uk/~spsolley -----------------------------------
_________________________________________________________________ It's Hotmail's 10th Birthday! Come and play Pass the Parcel http://www.msnpasstheparcel.com
==============================Original Headers============================== 14, 21 -- From hinmeigeng-at-hotmail.com Fri Dec 8 09:18:54 2006 14, 21 -- Received: from bay0-omc2-s37.bay0.hotmail.com (bay0-omc2-s37.bay0.hotmail.com [65.54.246.173]) 14, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB8FIrsx028913 14, 21 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 8 Dec 2006 09:18:53 -0600 14, 21 -- Received: from hotmail.com ([65.55.130.95]) by bay0-omc2-s37.bay0.hotmail.com with Microsoft SMTPSVC(6.0.3790.2668); 14, 21 -- Fri, 8 Dec 2006 07:18:53 -0800 14, 21 -- Received: from mail pickup service by hotmail.com with Microsoft SMTPSVC; 14, 21 -- Fri, 8 Dec 2006 07:18:52 -0800 14, 21 -- Message-ID: {BAY125-F15631D631515915344A32DCAD30-at-phx.gbl} 14, 21 -- Received: from 65.55.130.123 by by125fd.bay125.hotmail.msn.com with HTTP; 14, 21 -- Fri, 08 Dec 2006 15:18:46 GMT 14, 21 -- X-Originating-IP: [81.153.105.2] 14, 21 -- X-Originating-Email: [hinmeigeng-at-hotmail.com] 14, 21 -- X-Sender: hinmeigeng-at-hotmail.com 14, 21 -- From: "Robert H. Olley" {hinmeigeng-at-hotmail.com} 14, 21 -- To: Microscopy-at-MSA.Microscopy.Com 14, 21 -- Subject: Optical Physics 14, 21 -- Date: Fri, 08 Dec 2006 15:18:46 +0000 14, 21 -- Mime-Version: 1.0 14, 21 -- Content-Type: text/plain; format=flowed 14, 21 -- X-OriginalArrivalTime: 08 Dec 2006 15:18:52.0981 (UTC) FILETIME=[2B974650:01C71ADC] ==============================End of - Headers==============================
On Dec 7, 2006, at 3:59 PM, coridan-at-uiuc.edu wrote:
} 1) The plungerod sticks: On ths model, there is a horizontal } (left-to-right) adjustment to allow the operator to adjust the } guidance of the plungerod. However, it seems that some front-and-back } adjustments are required too. When we drop the plungerod, it is } sometimes slowed (or stopped middrop) by the sliding washer because } the rod is too far forward. } } 2) The foam base has cracked irreparably twice in the 2 months we have } been using it. This is a newer design for the vitrobot base: the foam } holds the LN2 and also insulates the brass ethane cup. We are using } it according to the directions, but this foam is breaking more } frequently than it should. } } I have contacted FEI, but I also thought I'd check to see if any other } Vitrobot users have experienced any of these problems. } Dear Rob, We have not had any problem with the plungerod on our Vitrobot, but we have had a problem with the cryogen cup occasionally moving forward even though the "semi-automated specimen transfer" box is not checked. This could cause the rod to become misaligned, but we have been lucky in that respect. I think our model does not have a plungerod adjustment that the user can operate. Several of our cryogen cups have cracked, so we always keep a spare. FEI is aware of the problem, and we are awaiting a solution. 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 Dec 8 11:29:09 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 kB8HT9rs021338 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 8 Dec 2006 11:29:09 -0600 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id CCD5710A2A2 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 8 Dec 2006 09:29:08 -0800 (PST) 4, 22 -- Received: from [192.168.159.233] (jpix-01.caltech.edu [131.215.2.133]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id E8BF510A2AE 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Fri, 8 Dec 2006 09:29:05 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200612072359.kB7Nx18t024356-at-ns.microscopy.com} 4, 22 -- References: {200612072359.kB7Nx18t024356-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {59df446f9d4362dde424aa7b07b4aac2-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: Vitrobot problems -- Base cracking? 4, 22 -- Date: Fri, 8 Dec 2006 09:35:13 -0800 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==============================
Most "frost free" freezers have a timer that controls a heating element. The heating element is turned on briefly which warms the freezer enough to melt the frost which has formed. A tube carries any melt water to an evaporation pan found usually under the freezer. This means that the temperature inside the freezer cycles through warmer and colder phases. This temperature cycling is probably the cause of the loss of enzyme activity. Some influenza vaccine manufacturers provide special containers for vaccines where frost free freezers will be used. These containers stabilize the temperature inside and prolong the vaccine's life. A similar insulated box would likely prolong the enzyme activity.
Bob Carter 2000 Bayshore Road Lopez Island, WA 98261-8595
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Hi,
I have 2 "blöde" questions, not directly related with microscopy, but perhaps one of you can still answer them:
1) I stored LM cryo-sections (unfixed, 5 µm thick) at -28°C for approx. 5 months and now I test the activity of alkaline phosphatase (very positive at the moment of cutting) I see no signal. Control show very strong signal, so it does not come from the technique. I wonder how I lost the enzyme activity. I know for proteins it is better to store à -80°C but I wouldn't expect to lose all the activity! The freezer is a "frost-free" freezer. Does someone know how these freezers work? Could the mechanism of frost-free freezers be detrimental to the enzymatic activity?
2) For the cell line OE21 in the ECACC catalog, I see "Hazard: CX". No other information! I couldn't find this cell line in the ATCC catalog and I couldn't find an explanation on the internet. What does it mean, CX?
Stephane
==============================Original Headers============================== 12, 32 -- From bob-at-rockisland.com Fri Dec 8 12:26:41 2006 12, 32 -- Received: from mars.rockisland.com (dnscache2.rockisland.com [64.119.0.12]) 12, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kB8IQeAA000754 12, 32 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Dec 2006 12:26:40 -0600 12, 32 -- Received: from localhost (localhost [127.0.0.1]) 12, 32 -- by localhost.rockisland.com (Postfix) with ESMTP id 66DC85E49 12, 32 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Dec 2006 10:26:40 -0800 (PST) 12, 32 -- (envelope-from bob-at-rockisland.com) 12, 32 -- X-Virus-Scanned: amavisd-new at rockisland.com 12, 32 -- Received: from mars.rockisland.com ([127.0.0.1]) 12, 32 -- by localhost (smtp.rockisland.com [127.0.0.1]) (amavisd-new, port 10024) 12, 32 -- with ESMTP id 4eiqsKXfgDiP for {Microscopy-at-microscopy.com} ; 12, 32 -- Fri, 8 Dec 2006 10:26:40 -0800 (PST) 12, 32 -- Received: from housecat (adsl-sj-15-153.rockisland.net [64.119.15.153]) 12, 32 -- by mars.rockisland.com (Postfix) with SMTP id 137035E47 12, 32 -- for {Microscopy-at-microscopy.com} ; Fri, 8 Dec 2006 10:26:40 -0800 (PST) 12, 32 -- (envelope-from bob-at-rockisland.com) 12, 32 -- Message-ID: {001901c71af6$6bcfb580$0302a8c0-at-housecat} 12, 32 -- From: "Bob Carter" {bob-at-rockisland.com} 12, 32 -- To: {Microscopy-at-microscopy.com} 12, 32 -- Subject: RE: 2 questions not directly relateed to microscopy 12, 32 -- Date: Fri, 8 Dec 2006 10:26:47 -0800 12, 32 -- MIME-Version: 1.0 12, 32 -- Content-Type: text/plain; 12, 32 -- format=flowed; 12, 32 -- charset="iso-8859-1"; 12, 32 -- reply-type=original 12, 32 -- Content-Transfer-Encoding: 8bit 12, 32 -- X-Priority: 3 12, 32 -- X-MSMail-Priority: Normal 12, 32 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 12, 32 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2962 ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (etienne.rivest-at-sympatico.ca) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, December 11, 2006 at 10:06:09 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both etienne.rivest-at-sympatico.ca as well as to the Microscopy Listserver ---------------------------------------------------------------------------
I am in the process of buying a microscope but, while browsing the web from ebay to majors manufacturers, I noted the wide variety of available items, specifications and price range. This poses some questions and I was glad to find your site that is the first ressource on my way that offers to give answers.
The first one is: what scope should I buy? Well, my goals in obtaining a microscope are primarely to observe potential bacterial activiy in home processed food preserves to establish preserving protocols (pressure canner sterelization, etc). The second is to observe beer yeasts in culture to determine its vitality (and purity of strain if possible). Lastly, to observe anything in the micro world for the only and pure fun of discovering hidden aspects of life and nature.
To clarify a bit, I would like to add that the more I read about microscopy (power range, types of eyepieces, objectives and condensers, numerical apperture, etc) the more I get confused. So, to be able to achieve my main expectations, what characteistics should I consider essential, considering that the microscope must also be financially affordable?
Thes last part of the preceding question leads to this one: beside the major makers who manufacture superb microscopes that I can in no way offer to myself, I saw lots of China (I guess) made kind of copies priced for less then half (when not the third) the price of what the majors ask. That's the case on ebay and a couple of sites I visited. Are those scopes adequate for private amateur home use? Would you consider it better not to have any microscope at all for a some years instead of having one of those in a few months or weeks?
Finally, what is the best way to find the microscope I need: new or used? popular sellers (like ebay)? Can you recommend a brand (or brands), a web site? I live in the country, relatively far from major cities, which is why I'm shoping through the web.
I thank you very much to offer the opportunity to ask you my questions. Thank you in advance for your help.
I am trying to help a colleague with autoradiography on a cancer project. I was intending to cut t 0.5 - 1.0 um cross sections of the tumors after embedding them in BMMA (butylmethylmethacrylate) which is a lot like JB-4 but easily extractable. I just learned that the tumors are bigger than I expected - between 1 and 2 cm. I need to cut the full width cross section since we are looking for distribution of a drug. I have always cut methacrylate resins on a standard ultramicrotome. I have heard but not seen a so-called JB-4 microtome. Is this a standard microtome with a metal blade or is it different? The lab I will be working in can get a Reichert Supercut 2050 microtome to use on this project. I see literature references where this microtome was used to cut JB-4. Is anyone familiar with this model? Is it a standard microtome with either disposable razor blade or paraffin microtome blade? Does anyone know how big (wide) a block you can cut on a standard microtome? Thanks, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Has anyone cleaned the oil out of an oil mist filter from a rotary pump from an Hitachi TEM? Or any similar filter? The pump uses mineral oil. The instructions say to use Perkleen or similar defatting agent; don't use solvents. However I've never heard of Perkleen and have no idea what might be in it. Any suggestions?
Diana
==============================Original Headers============================== 3, 18 -- From dianavd-at-eye.usyd.edu.au Mon Dec 11 16:09:41 2006 3, 18 -- Received: from galen.med.usyd.edu.au (nugalen.med.usyd.edu.au [129.78.36.39]) 3, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBBM9fo9014444 3, 18 -- for {Microscopy-at-microscopy.com} ; Mon, 11 Dec 2006 16:09:41 -0600 3, 18 -- Received: from [129.78.203.144] (helo=[129.78.203.144]) 3, 18 -- by galen.med.usyd.edu.au with esmtp (Exim 4.43) 3, 18 -- id 1GttL9-0001tc-Ts 3, 18 -- for Microscopy-at-microscopy.com; Tue, 12 Dec 2006 09:09:40 +1100 3, 18 -- Mime-Version: 1.0 (Apple Message framework v752.2) 3, 18 -- Content-Transfer-Encoding: 7bit 3, 18 -- Message-Id: {15DFD0A3-0912-454D-A749-E69250CC17C1-at-eye.usyd.edu.au} 3, 18 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 3, 18 -- To: Microscopy {Microscopy-at-microscopy.com} 3, 18 -- From: Diana van Driel {dianavd-at-eye.usyd.edu.au} 3, 18 -- Subject: cleaning oil mist filter 3, 18 -- Date: Tue, 12 Dec 2006 09:09:34 +1100 3, 18 -- X-Mailer: Apple Mail (2.752.2) 3, 18 -- X-Spam-Score: -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 qfxingtem-at-hotmail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Title-Subject: [Filtered] High Tension Stability of Tecnai-F20ST
Question: Dear Listers:
Does anyone know the high-tension stability (change of the accelerating voltage versus time) of Tecnai-F20ST after setting to the voltage for 3 hours? I used 120kV. The nominal value at 200kV is aslo okay.
O} } Hi all, } } Has anyone cleaned the oil out of an oil mist filter from a rotary } pump from an Hitachi TEM? Or any similar filter? The pump uses } mineral oil. The instructions say to use Perkleen or similar } defatting agent; don't use solvents. However I've never heard of } Perkleen and have no idea what might be in it. Any suggestions? } } Diana }
I would guess that it's a trade name for perchloroethylene, which has been and still may be being used as a drycleaning solvent. It is, of course, a solvent, heaven only knows what they mean by 'don't use a solvent'. Like all halogenated solvents, it isn't a nice thing to absorb through the skin of your hands or your lungs.
cheers
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
==============================Original Headers============================== 6, 27 -- From r.sims-at-auckland.ac.nz Mon Dec 11 19:24:38 2006 6, 27 -- Received: from chico.itss.auckland.ac.nz (chico.itss.auckland.ac.nz [130.216.190.12]) 6, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBC1OaQa015014 6, 27 -- for {Microscopy-at-microscopy.com} ; Mon, 11 Dec 2006 19:24:37 -0600 6, 27 -- Received: from localhost (localhost.localdomain [127.0.0.1]) 6, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id 0D70C345B6; 6, 27 -- Tue, 12 Dec 2006 14:24:33 +1300 (NZDT) 6, 27 -- Received: from chico.itss.auckland.ac.nz ([127.0.0.1]) 6, 27 -- by localhost (smtpb.itss.auckland.ac.nz [127.0.0.1]) (amavisd-new, port 10024) 6, 27 -- with ESMTP id 10372-02; Tue, 12 Dec 2006 14:24:32 +1300 (NZDT) 6, 27 -- Received: from rs (r.sims.glg.auckland.ac.nz [130.216.59.18]) 6, 27 -- by chico.itss.auckland.ac.nz (Postfix) with ESMTP id D96C2343F3; 6, 27 -- Tue, 12 Dec 2006 14:24:31 +1300 (NZDT) 6, 27 -- From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} 6, 27 -- To: dianavd-at-eye.usyd.edu.au 6, 27 -- Date: Tue, 12 Dec 2006 14:29:37 +1300 6, 27 -- MIME-Version: 1.0 6, 27 -- Subject: Re: [Microscopy] cleaning oil mist filter 6, 27 -- Cc: Microscopy-at-microscopy.com 6, 27 -- Message-ID: {457EBCD1.25576.174E2CD-at-localhost} 6, 27 -- Priority: normal 6, 27 -- In-reply-to: {200612112210.kBBMAbnJ016111-at-ns.microscopy.com} 6, 27 -- X-mailer: Pegasus Mail for Windows (4.21c) 6, 27 -- Content-type: text/plain; charset=US-ASCII 6, 27 -- Content-transfer-encoding: 7BIT 6, 27 -- Content-description: Mail message body 6, 27 -- X-Virus-Scanned: by amavisd-new at mailhost.auckland.ac.nz ==============================End of - Headers==============================
2050s are heavy duty motorised microtomes designed to cope with the high cutting forces associated with resin sectioning so should suit your purpose well, assuming you can accommodate a glass, diamond or a tungsten carbide blade. They can also accomodate standard wax sectioning steel and disposable blades. All of these options depend on the type of knife stage your 2050 is fitted with.
I don't think that standard blades would stand up to sectioning large MMA blocks, but listers may know otherwise.
I regularly use a 2050 with glass triangular knives with a cutting edge the thickness of the glass (1cm) to cut GMA and MMA blocks 1-2 cm long - positioning the block with the narrow edge meeting the knife and with its leading edge trimmed to a point. This allows a gradual introduction of the cutting force, and gives you something to pick up with fine forceps without damaging the tissue if cutting the sections dry.
Tungsten carbide blades also work well for larger blocks. I have found that MMA sections best if you wet the block and blade with 70% EtOH and smooth the section flat with a fine artist brush before transferring to a water bath for collecting onto slides.
As MMA sections can be floated direct onto water, we use a histodiamond to provide best quality sections having first trimmed in with a glass or TC blade.
Get in touch if you need further info.
Good luck,
Alastair
At 15:23 11/12/2006 -0600, you wrote:
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Alastair McKinnon Histology & EM Facility Manager University of Aberdeen, Institute of Medical Science Foresterhill, Aberdeen, AB25 2ZD tel: +44(0) 1224 552923 (office); +44(0) 1224 555911 (lab) fax: +44(0) 1224 559533; email: a.d.mckinnon-at-abdn.ac.uk www.abdn.ac.uk/ims/h-em
==============================Original Headers============================== 16, 18 -- From a.d.mckinnon-at-abdn.ac.uk Tue Dec 12 03:19:20 2006 16, 18 -- Received: from mailhub2.abdn.ac.uk (mailhub2.abdn.ac.uk [139.133.7.24]) 16, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBC9JJGd004211 16, 18 -- for {Microscopy-at-microscopy.com} ; Tue, 12 Dec 2006 03:19:20 -0600 16, 18 -- Received: from clsm-002230.ims.abdn.ac.uk ([139.133.169.205] helo=CLSM-002230.abdn.ac.uk) 16, 18 -- by mailhub2.abdn.ac.uk with esmtp (Exim 4.52) 16, 18 -- id 1Gu3nC-0007hi-Fv; Tue, 12 Dec 2006 09:19:19 +0000 16, 18 -- Message-Id: {5.2.1.1.0.20061212084505.011d2040-at-mailms.abdn.ac.uk} 16, 18 -- X-Sender: pat081-at-mailms.abdn.ac.uk 16, 18 -- X-Mailer: QUALCOMM Windows Eudora Version 5.2.1 16, 18 -- Date: Tue, 12 Dec 2006 09:19:15 +0000 16, 18 -- To: phillipst-at-missouri.edu 16, 18 -- From: Alastair McKinnon {a.d.mckinnon-at-abdn.ac.uk} 16, 18 -- Subject: Re: [Microscopy] JB-4, BMMA, Supercut 2050 microtome questions 16, 18 -- Cc: Microscopy-at-microscopy.com 16, 18 -- In-Reply-To: {200612112123.kBBLNsbs010977-at-ns.microscopy.com} 16, 18 -- Mime-Version: 1.0 16, 18 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
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To submit your abstract, review submission guidelines, and learn more about the conference, visit www.veeco.com/nanoconference
Take part as a presenter in the industry's most dynamic conference!
==============================Original Headers============================== 14, 20 -- From MCarlyle-at-veeco.com Tue Dec 12 09:27:04 2006 14, 20 -- Received: from sboexch2.int.veeco.com (SBOEXCH2.VEECO.com [68.111.35.167]) 14, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBCFR3FW031607 14, 20 -- for {Microscopy-at-Microscopy.Com} ; Tue, 12 Dec 2006 09:27:03 -0600 14, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 14, 20 -- content-class: urn:content-classes:message 14, 20 -- MIME-Version: 1.0 14, 20 -- Content-Type: text/plain; 14, 20 -- charset="utf-8" 14, 20 -- Subject: AFM -- Seeing at the Nanoscale V Conference Call for Papers 14, 20 -- Date: Tue, 12 Dec 2006 07:27:02 -0800 14, 20 -- Message-ID: {625BD6DD96DE554DBB0558FFE70E0F420120016E-at-sboexch2.int.veeco.com} 14, 20 -- X-MS-Has-Attach: 14, 20 -- X-MS-TNEF-Correlator: 14, 20 -- Thread-Topic: AFM -- Seeing at the Nanoscale V Conference Call for Papers 14, 20 -- Thread-Index: AcceAfjYH90V77/ITkSawDkMxJD/Xg== 14, 20 -- From: "Marlene Carlyle" {MCarlyle-at-veeco.com} 14, 20 -- To: "Microscopy-at-Microscopy.Com" {'Microscopy-at-Microscopy.Com'} 14, 20 -- Content-Transfer-Encoding: 8bit 14, 20 -- X-MIME-Autoconverted: from base64 to 8bit by ns.microscopy.com id kBCFR3FW031607 ==============================End of - Headers==============================
---------------------------------------------------------------------- FOCUS ON MICROSCOPY 2007 -- Valencia, Spain -- April 10-13, 2007 ----------------------------------------------------------------------
20th International Conference on 3D Image Processing in Microscopy 19th International Conference on Confocal Microscopy
Dear Colleagues,
January 15, 2007, the deadline for abstract submission for the Valencia FOM2007 conference is nearing. Please submit your abstract by that date.
The program for the conference will be finalized and available on our website on Feb. 6, 2007. Authors will 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.
After the successful FOM2006 conference held in Perth in April this year, the next conference: Focus on Microscopy 2007 will take place in Valencia, Spain from Tuesday April 10 to Friday April 13th, 2007. Please note that this is in the week after Easter 2007. 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 Valencia.
Focus on Microscopy 2007 is the continuation of a conference series, started in 1988, presenting the latest innovations in optical microscopy and their application in biology, medicine and the material sciences. Key subjects for the conference are the theory and practice of 3D optical imaging, related 3D image processing, together with reporting on the ever increasing spatial resolutions and sophisticated imaging modes coming available in sectioning microscopy.
The conference series is in addition known for covering the rapid development of advanced fluorescence labeling techniques for confocal and multi-photon 3D imaging of -live- biological specimens. Laser light in combination with 3D microscopy is starting to play an increasingly important role as a tool at the sub micrometer scale in cell biology for dissection and isolation of structures of interest for subsequent analysis. Specific sessions on this subject and advances in endoscopic and non-invasive imaging are planned for FOM2007.
Important dates: Deadline for the submission of abstracts: January 15, 2007 Acceptance of contributions, draft program: February 6, 2007 Deadline for early registration: February 26, 2007
To stay informed about the conference please leave your name and email at http://www.FocusOnMicroscopy.org/stayinformed
Welcoming you to Valencia for the FOM2007 conference and exhibition, on behalf of the organising committee,
- Manuel Martinez-Corral, University of Valencia, Spain - Fred Brakenhoff, University of Amsterdam, The Netherlands -- E-mail: info2007-at-FocusOnMicroscopy.org Web: www.FocusOnMicroscopy.org
==============================Original Headers============================== 16, 27 -- From brakenho-at-science.uva.nl Tue Dec 12 10:30:07 2006 16, 27 -- Received: from imap.science.uva.nl (imap.science.uva.nl [146.50.4.51]) 16, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBCGU61b011653 16, 27 -- for {microscopy-at-msa.microscopy.com} ; Tue, 12 Dec 2006 10:30:07 -0600 16, 27 -- Received: from webmail.science.uva.nl [146.50.4.91] 16, 27 -- by imap.science.uva.nl with ESMTP (sendmail 8.11.6p2/config 11.38). 16, 27 -- id kBCGU5M11482; Tue, 12 Dec 2006 17:30:05 +0100 16, 27 -- X-Organisation: Faculty of Science, University of Amsterdam, The Netherlands 16, 27 -- X-URL: http://www.science.uva.nl/ 16, 27 -- Received: from localhost ([127.0.0.1]) 16, 27 -- (SquirrelMail authenticated user brakenho) 16, 27 -- by webmail.science.uva.nl with HTTP; 16, 27 -- Tue, 12 Dec 2006 17:30:05 +0100 (CET) 16, 27 -- Message-ID: {50121.145.18.160.47.1165941005.squirrel-at-webmail.science.uva.nl} 16, 27 -- Date: Tue, 12 Dec 2006 17:30:05 +0100 (CET) 16, 27 -- Subject: FOM2007 Abstract deadline 15 Jan. Valencia, Spain, April 10-13, 16, 27 -- Focus on Microscopy, Registration 16, 27 -- From: "Fred Brakenhoff" {brakenho-at-science.uva.nl} 16, 27 -- To: microscopy-at-msa.microscopy.com 16, 27 -- Cc: brakenhoff-at-science.uva.nl 16, 27 -- User-Agent: SquirrelMail/1.4.6-rc1 16, 27 -- MIME-Version: 1.0 16, 27 -- Content-Type: text/plain;charset=iso-8859-1 16, 27 -- Content-Transfer-Encoding: 8bit 16, 27 -- X-Priority: 3 (Normal) 16, 27 -- Importance: Normal 16, 27 -- X-Virus-Scanned: by amavisd-new ==============================End of - Headers==============================
The JB-4 is a retracting motorized microtome designed for semi-thin sections. Depending upon the available knife holders and your sample, it will use disposable steel, triangular glass, Ralph knives, tungsten carbide or diamond knives. We've used glass knives and Histodiamonds with ours to cut epoxy and methacrylate sections, 0.5 to 5 µm thick. As, Alastair says, a razor blade or a standard steel blade won't survive what you want to cut. That plastic will curl the edge or even pull out the metal.
Regards, Glen } } } --------------------------------------------------------------------- } } ------- } } 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 am trying to help a colleague with autoradiography on a cancer } } project. I } } was intending to cut t 0.5 - 1.0 um cross sections of the tumors } } after } } embedding them in BMMA (butylmethylmethacrylate) which is a lot } } like JB-4 } } but easily extractable. I just learned that the tumors are bigger } } than I } } expected - between 1 and 2 cm. I need to cut the full width cross } } section } } since we are looking for distribution of a drug. I have always cut } } methacrylate resins on a standard ultramicrotome. I have heard } } but not } } seen a so-called JB-4 microtome. Is this a standard microtome } } with a metal } } blade or is it different? The lab I will be working in can get a } } Reichert } } Supercut 2050 microtome to use on this project. I see literature } } references } } where this microtome was used to cut JB-4. Is anyone familiar } } with this } } model? Is it a standard microtome with either disposable razor } } blade or } } paraffin microtome blade? Does anyone know how big (wide) a block } } you can } } cut on a standard microtome? 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============================== 4, 27 -- From glenmac-at-u.washington.edu Tue Dec 12 12:42:42 2006 4, 27 -- Received: from mxout3.cac.washington.edu (mxout3.cac.washington.edu [140.142.32.166]) 4, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBCIgg5j025805 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 12 Dec 2006 12:42:42 -0600 4, 27 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.32.139]) 4, 27 -- by mxout3.cac.washington.edu (8.13.7+UW06.06/8.13.7+UW06.09) with ESMTP id kBCIgf7j004248 4, 27 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 12 Dec 2006 10:42:41 -0800 4, 27 -- X-Auth-Received: from [128.95.178.71] ([128.95.178.71]) 4, 27 -- (authenticated authid=glenmac) 4, 27 -- by smtp.washington.edu (8.13.7+UW06.06/8.13.7+UW06.09) with ESMTP id kBCIgfPG001114 4, 27 -- (version=TLSv1/SSLv3 cipher=AES128-SHA bits=128 verify=NOT) 4, 27 -- for {microscopy-at-microscopy.com} ; Tue, 12 Dec 2006 10:42:41 -0800 4, 27 -- Mime-Version: 1.0 (Apple Message framework v752.2) 4, 27 -- In-Reply-To: {200612120921.kBC9LRph006717-at-ns.microscopy.com} 4, 27 -- References: {200612120921.kBC9LRph006717-at-ns.microscopy.com} 4, 27 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 4, 27 -- Message-Id: {749F3D15-7E41-447D-AB21-DFFECCAE58B7-at-u.washington.edu} 4, 27 -- From: Glen MacDonald {glenmac-at-u.washington.edu} 4, 27 -- Subject: Re: [Microscopy] Re: JB-4, BMMA, Supercut 2050 microtome questions 4, 27 -- Date: Tue, 12 Dec 2006 10:42:35 -0800 4, 27 -- To: "ListServer-at-MSA.Microscopy.Com Listserver" {microscopy-at-microscopy.com} 4, 27 -- X-Mailer: Apple Mail (2.752.2) 4, 27 -- X-PMX-Version: 5.2.2.285561, Antispam-Engine: 2.5.0.283055, Antispam-Data: 2006.12.12.102932 4, 27 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P1_5 0, __C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __FRAUD_419_BADTHINGS 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' 4, 27 -- Content-Transfer-Encoding: 8bit 4, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBCIgg5j025805 ==============================End of - Headers==============================
Thanks for all the recent advice on my question on the 2050 microtome and cutting JB-4 and BMMA. Today's question concerns squirt bottles for delivering ethanol (50, 70, 95 and 100%) to vials during the dehydration step prior to infiltration in resin. I have a set of squirt bottles with locking seals that are on their last legs and need to break down and get a net set. Does anyone know of an especially good type of squirt bottle that keeps the ethanol series from taking in water from the atmosphere? And yes, I know all about using dry ethanol and sieves, etc and that isn't part of this question! Thanks in advance, Tom
Thomas E. Phillips, PhD Professor of Biological Sciences Director, Molecular Cytology Core 2 Tucker Hall University of Missouri Columbia, MO 65211-7400
Hello, We are still using Vestopal W for embedding of our samples. During the years well polymerized blocks were transparent pink (aprox. color: #ffc0cb in hexadecimal) but now the polymerized block are transparent khaki to green. We have bought new tertiary butyl perbenzoate and cobalt naphthenate but the new mixture of Vestopal again polymerized in transparent khaki to green color. We have been using standard protocol temperature curring (60 oC) for years and we have never before observed such change in color of polymerized blocks. Please, could anybody explain it?
Thanking you in advance Oldrich
------------------------------------------ Oldrich Benada Institute of Microbiology, Acad. Sci. CR Videnska 1083 142 20 Praha 4 - Krc Czech Republic
==============================Original Headers============================== 4, 20 -- From benada-at-biomed.cas.cz Wed Dec 13 03:00:21 2006 4, 20 -- Received: from biomed.cas.cz (mail2.biomed.cas.cz [147.231.40.32]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBD90KX1000564 4, 20 -- for {microscopy-at-microscopy.com} ; Wed, 13 Dec 2006 03:00:21 -0600 4, 20 -- Received: from [147.231.44.101] (u117ob.mbu.cas.cz [147.231.44.101]) 4, 20 -- by biomed.cas.cz (8.13.3+Sun/8.13.3) with ESMTP id kBD8xGsO004489 4, 20 -- for {microscopy-at-microscopy.com} ; Wed, 13 Dec 2006 09:59:16 +0100 (CET) 4, 20 -- From: benada-at-biomed.cas.cz 4, 20 -- To: microscopy-at-microscopy.com 4, 20 -- Date: Wed, 13 Dec 2006 10:00:16 +0100 4, 20 -- MIME-Version: 1.0 4, 20 -- Subject: Vestopal W question 4, 20 -- Message-ID: {457FCF30.29743.49651F-at-benada.biomed.cas.cz} 4, 20 -- Priority: normal 4, 20 -- X-mailer: Pegasus Mail for Windows (4.41) 4, 20 -- Content-type: text/plain; charset=US-ASCII 4, 20 -- Content-transfer-encoding: 7BIT 4, 20 -- Content-description: Mail message body 4, 20 -- X-Antivirus: avast! (VPS 0657-2, 12.12.2006), Outbound message 4, 20 -- X-Antivirus-Status: Clean ==============================End of - Headers==============================
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I've recently come into possesion of an old EM201. I was wondering if anyone out there has a PDF of the Installations manual (I have the other three, including Preinstallation). Also, does anyone know if the chilled water line can be filled with anything but water? I was wondering about an Ethanol solution or glycol (something with a depressed freezing point). It will be in a room that might get down to freezing temperatures if the electricity happens to fail. Will the seals stand up to chemicals other than water? Are there any other considerations for the cooling loop? Any help would be greatly appreciated. Thanks.
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Email: ht-at-nhm.ac.uk Name: Harry Taylor
Organization: NHM
Title-Subject: [Filtered] Wild M410 macroscope
Question: Hi there, Just wondering how easy it is to convert a Wild M410 macroscope body into the M420? I believe I just need to add the internal photo prism (my 410 already has the photo tube fitted) Does anybody know the part number of this item?......thanks in advance!
We get asked this fairly regularly by schools and parents (so we have stock replies). Often professional microscopists aren't the best source of information for cheap microscope's as all our optical systems are always over £10,000, and most lab mainstays like our three confocal microscopes come in at nearer £200,000 each (and users still complain about image quality). I assume you are asking about home use.
Cheap microscopes under £100 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 and you may make an expensive mistake. Look for old branded 'laboratory' microscopes e.g. Bausch & Lomb, Prior, Leitz, Reichert, Baker, but not Tasco toys. Famous existing brands like Zeiss and Olympus attract a high premium. The sellers are often not microscopists though, and many are sold as collector's items and not intended for 'scientific' use. However many home users often get bored with new microscopes after a while, so there is a second hand market for cheaper school/college lab grade models. Also I would definitely try any local microscope enthusiast clubs - they aren't as common as the many excellent astronomy [telescope] clubs but they are about and have knowledgeable enthusiasts with an eye for low cost quality systems.
There are suppliers geared up to providing cheaper microscopes for schools & colleges, so you can ask around at school/college's science departments, but expect to pay nearer £500 each for a minimum quality setup (although with those like bottom end Meade [www.meade.com] at around £100 you can see something at low mag (~20x objective i.e. around 180x mag) with a quality stained section.
For pond life etc. a stereoscopic 'dissecting' microscope (40x to 120x mag) is ideal for home use. Also remember of course that you can get a really long way with a good large magnifying glass (not the really small hi-mag cheap folding lens ones, try before you buy) - I have a few excellent ones at home for £1 and a good low mag Osram one that includes an illuminating halogen bulb at £8. In general I would say a well made stereo dissecting microscope is a good choice (if not the best) for kids as it's great for viewing living things and enlarges what you can see already - look for 40x rather than 4x though. They are a bit expensive (£500+) though so you would probably have to buy secondhand (look out for branded ones like Bausch & Lomb, and Prior). These are ideal for colonies but unsuitable for viewing single cells where something like 400x to 600x is preferred (expensive 40 to 63x objectives), and requires a standard compound microscope.
Generally prepared slides can rapidly get very boring for all ages, but living or unusual things (even hamster fur) always attract an audience. Also try your flatbed film scanner, (not the LiDe types that have a very limited depth of focus, and any from around £60 upwards) that will be good for looking at soft static things: leaves, fruit, nuts, household objects (scan at max resolution and try reflected and 'film' mode). In the UK there are sites like http://www.wedgwood-group.com/microscopes_digital.htm that cater for schools and colleges, providing standard compound and stereo microscopes as well as cheap PC video based microscope solutions where the whole class of children can look at a computer screen with some pushing and shoving. Best to try them on 'approval' as many cheap microscopes can be very disappointing if you expect too much. For your use (living cells) you will probably want some form on contrast enhancement like Phase Contrast optics that adds to the complexity and expense (and starts to put the microscope in the £1,000+ category). Fixing & staining cells obviously reduces the need for any optical contrast enhancement.
Excellent pre-prepared stained slides of simple organisms, plant stems and leaves or bits of rats, insects etc.. can be bought via ebay, but they tend to be expensive and are easily broken by any age-group. Mounted slides keep well though, so 'vintage' ones even from 50 years ago can still look OK. Again school suppliers are geared up to provide for this sort of thing cheaply.
At home and our Primary school (under 11) we use the Digital Blue QX-5 (£70) - it's fun but pretty useless for serious microscope work as it's so low res (but at 640 x 480 better than the old Intel QX-3 it replaced). It puts the image on a PC screen. I have one at home for my kids (boy 10 and girl 12) but it only gets occasional use now the novelty has worn off. See http://micro.magnet.fsu.edu/optics/intelplay/index.html (not updated for the QX-5 but all applies - the site even discusses ways of contrast enhancement etc..). Once on the PC the 640x480 images can be manipulated and pasted etc, and the QX-5 does time-lapse for things like crystal growth. Living plants growing and small animals. This toy has nowhere near the resolution you require though. The similar but far better built Olympus MIC-D was great but being over £500 it was just too expensive for most schools and is now discontinued - there are other similar budget video microscope systems about though.
The macro on a good digital camera (like the image stabilised 5MP Canon S2IS - http://www.dpreview.com/reviews/canons2is)is also worth a try, particularly with a small tripod and halogen bendy desk lamp, if very close-up. I have an E500 digital SLR system with enlargement rings but they are more difficult to use. You can get quite reasonable pictures by resting a small compact digital camera lens against the eyepiece of a microscope. Plus you can use the camera for normal photography when you get bored with microscopy.
For details on your interest try books like the Bioaerosols Handbook by C.S. Cox. Granted this is for sampling bioaerosols (bacteria etc.. floating in air) not food as in your interest, but their must be similar books about for that. I mention the handbook as I wrote the chapter on microscopy and image analysis so I am very familiar with the book.
By the way, if you get a microscope, do try growing crystals on a slide, a few drops of a saturated solution of salt (NaCl) or copper sulphate will grow superb crystals on the surface of a slide when viewed under a microscope (but it takes a few hours for the crystals to form and they often look best before the liquids all gone). Just make sure you don't drive the objective tips into the solution. It's not biology but its fun (see http://micro.magnet.fsu.edu/.
Sorry I can't offer more specific help as our microscopes are rather more expensive than the one you wish to track down.
Regards
Keith
Try looking in Amazon.com for decent microscope books with lots of pictures - and they have a good customer review system for books and even microscopes (often viewing micrographs in books is better than trying to see it yourself down a microscope). Plus try web searches for general sites like these (and for more specific topics) :
http://www.101science.com/Microscope.htm http://micro.magnet.fsu.edu/ http://www.microscopy-uk.org.uk/index.html http://www.btinternet.com/~stephen.durr/ http://www.mccroneatlas.com http://www.diatoms.co.uk [for fun images made of diatoms]
Examples of sources for cheap school grade microscopes in the UK are:
Have an internet search for school/lab suppliers in Canada.
-----Original Message----- X-from: etienne.rivest-at-sympatico.ca [mailto:etienne.rivest-at-sympatico.ca] Sent: 11 December 2006 17:19 --------------------------------------------------------------------------- Please reply to both etienne.rivest-at-sympatico.ca as well as to the Microscopy Listserver ---------------------------------------------------------------------------
I am in the process of buying a microscope but, while browsing the web from ebay to majors manufacturers, I noted the wide variety of available items, specifications and price range. This poses some questions and I was glad to find your site that is the first ressource on my way that offers to give answers.
The first one is: what scope should I buy? Well, my goals in obtaining a microscope are primarely to observe potential bacterial activiy in home processed food preserves to establish preserving protocols (pressure canner sterelization, etc). The second is to observe beer yeasts in culture to determine its vitality (and purity of strain if possible). Lastly, to observe anything in the micro world for the only and pure fun of discovering hidden aspects of life and nature.
To clarify a bit, I would like to add that the more I read about microscopy (power range, types of eyepieces, objectives and condensers, numerical apperture, etc) the more I get confused. So, to be able to achieve my main expectations, what characteistics should I consider essential, considering that the microscope must also be financially affordable?
Thes last part of the preceding question leads to this one: beside the major makers who manufacture superb microscopes that I can in no way offer to myself, I saw lots of China (I guess) made kind of copies priced for less then half (when not the third) the price of what the majors ask. That's the case on ebay and a couple of sites I visited. Are those scopes adequate for private amateur home use? Would you consider it better not to have any microscope at all for a some years instead of having one of those in a few months or weeks?
Finally, what is the best way to find the microscope I need: new or used? popular sellers (like ebay)? Can you recommend a brand (or brands), a web site? I live in the country, relatively far from major cities, which is why I'm shoping through the web.
I thank you very much to offer the opportunity to ask you my questions. Thank you in advance for your help.
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Email: twigg-at-estd.nrl.navy.mil Name: Mark Twigg
Organization: Naval Research Laboratory
Title-Subject: [Filtered] Epoxy
Question: Generally my favorite epoxy for gluing down TEM specimens is Epotek H20E Silver Epoxy. This epoxy requires curing at about 120C, which is fine for most of my specimens. What I especially like about this epoxy is that it has a slightly pasty consistency, so that it does not suck the specimen in using capillary action. What I want to know is if there is another slightly pasty epoxy or glue out there which cures at room temperature. Some samples cannot endure curing at 120C, so it would be good have know of an alternative to Epotek H20E.
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Email: vchristie-at-altransolutions.com Name: Van Christie
Organization: Altran Solutions
Title-Subject: [Filtered] Zeiss vs JEOL
Question: We are ready to pull the trigger on a new VP SEM with an EDS system. I have decided on a Thermo EDS as I've had good experience with them. But the EM is a mystery. Both have great capabilities and similar features. The Zeiss is my first choice as in the VP mode, they have a vp secondary detector that will work with the BS detector. Four main boards control the whole instrument. One simple roughing pump, only a couple of valves and a turbo pump for HV.
So does anyone have experience with either of these newer machines? Any help is appreciated.
I am running a Zeiss EVO 40 with VPSE detector and am VERY happy with it (no analytical set-up). Our application is biological samples in biodiversity research (shells, tissues, resin casts, insects). When we wrote the NSF-MRI grant, I looked also at JEOL. JEOL came in second after the other common vendors for the following reasons. I selected the Zeiss for some of the reasons outlined below.
I do like the flexibility and ease of switching between modes, and that all variables can be continuously adjusted (15.00 kV or 15.01 kV is your choice, it is not 15 or 16 kV, and that for every parameter). Signal mixing (also continuous) is a great asset. I often do a 70-80% SE with 20-30% of 2 quadrants of BSE for "counter illumination".
No cooler required is another plus, just one less piece of equimpment to take care of. You also plug the instrument simply into a 110 outlet, no need getting 220/240V lines installed. So apart of the 4 boards, yet another way how the Zeiss is simple and robust.
Filament life is very good. If you check saturation about once a week, you can easily get 300-400 h out of a filament.
Macroprogramming is feasible, but not terribly intuive, but it can be done. I have some custom one-click-buttons for image taking (line integration, set of frame integration parameter pairs) with subsequent saving. Also did for one user a pull down-menue with standard magnifications.
Do get the keyboard with the adjustment buttons on it. You can then focus and magnify etc. at the same time. We do a bunch of student training and have visiting scientists, and the button-key-board makes it easier to get into the instrument.
Problems? We've had it running for 2 years with no significant down-time.
If you have any question please feel free to check back.
best wishes
Daniel
************************************************* Daniel L. Geiger, Ph.D. Research Curator of Electron Microscopy Santa Barbara Museum of Natural History - Invertebrate Zoology 2559 Puesta del Sol Road Santa Barbara, CA 93105 USA
On Thu Dec 14 5:47 , vchristie-at-altransolutions.com sent:
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Email: tomw-at-uidaho.edu Name: Tom Williams
Organization: U of Id
Title-Subject: [Filtered] Cleaning a Lab6 emitter
Question: I am running a Denka Lab6 on an AMRAY 1830. We a lot of samples mounted in epoxy and the LaB6 now has a bad coating of what I think is epoxy residue (visible under a binoc). Does anyone have a suggestion on how to clean this safely?
You may use any organic solvent that is not corrosive to metals, Freon-113 or Chloroform are very good, tetrachloromethane is good, or any other chlorinated hydrocarbon if you can get any- these are usually restricted. Just soak the emitter, do not sonicate!
I dealt with a number oxidized LaB6 cathodes, when for example a user accidentally inserted specimen holder in the TEM without pre-pumping, while LaB6 was hot (some did so many times in row :-) Then vacuum system shots down, etc., and LaB6 catches some O2 from the air. All I do to cure that condition is to run LaB6 (for a day or so) at a slightly higher temperature at low KV and low emission to let poisoned layer of the crystal to evaporate. It usually works, but then there is a limit of how much abuse LaB6 can take.
However, I have doubts about the very cause of your LaB6 problem. What is the history of symptoms?
First, how did contamination got in the gun area? An SEM equipped for LaB6 use must be pumped differentially - the gun and column have dedicated IGP and are separated by differential aperture from the specimen chamber.
Second, I can hardly imagine organics condensing on an object heated to about 2000 degrees F.
History of the symptoms?
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: {tomw-at-uidaho.edu} To: {vitalylazar-at-att.net} Sent: Thursday, December 14, 2006 7:13 PM
I'm not sure about how to clean the LaB6 from Denka. But I have done this once with FEI cathodes.
What is puzzling is why you have to do this at all. The 1830 if set up for LaB6 has a 30L/m ion pump for the gun chamber. This should and does keep gun chamber vacuum really good. What IPG value are you currently reading? If you do not have a gun chamber ion pump, you cannot run LaB6. If you do have it, there is no obvious reason why the LaB6 would be contaminated. It makes no sense to me. I never had this happen. IPG was typically 40uA. Hopefully, your stand pipe valve is closed.
gary g.
At 04:13 PM 12/14/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Thu Dec 14 19:55:14 2006 10, 20 -- Received: from qsmtp4.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kBF1tDLR029343 10, 20 -- for {microscopy-at-microscopy.com} ; Thu, 14 Dec 2006 19:55:13 -0600 10, 20 -- Received: (qmail 3838 invoked from network); 14 Dec 2006 17:50:37 -0800 10, 20 -- Received: by simscan 1.1.0 ppid: 3786, pid: 3833, t: 0.1689s 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; 14 Dec 2006 17:50:36 -0800 10, 20 -- Message-Id: {7.0.1.0.2.20061214175110.02568880-at-gaugler.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 20 -- Date: Thu, 14 Dec 2006 17:55:08 -0800 10, 20 -- To: tomw-at-uidaho.edu 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] viaWWW: Cleaning a Lab6 emitter 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200612150013.kBF0DT36008436-at-ns.microscopy.com} 10, 20 -- References: {200612150013.kBF0DT36008436-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset=us-ascii; format=flowed; x-avg-checked=avg-ok-20BE2D9D ==============================End of - Headers==============================
As we'll cecome next year a new TEM, we are studying the question to move all our microscopes in an other part of the building. But, if the place seems to have interesting side, obviously something must be wrong ! There is a power line which runs in a technical corridor in the underground, carrying some 800-1000 Amps for the supply of the whole building, and generating something like 50 mG in one of the possible room.
So, I have three questions :
First, is it possible to shield such a power line, to lower the field at it source (1" thick aluminum or cooper, or so) ? Is such a shielding technically possible, and not too expensive ? I think it would be better to try first to limit the perturbation at it source !
Secondly, what are the feedback from people working with dynamic magnetic field compensation ? How much do you have without it, and how does it work, with such a big field. Does it react fast enough, when the stray field changes.
Thirdly, a question for europeen lister, I'm looking for europeen manufactuerer of such dynamic compensation and/or static shielding. I know some, but I want to have several contacts.
Thanks in advance for help.
-- 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
} Question: I am running a Denka Lab6 on an AMRAY 1830. We a } lot of samples mounted in epoxy and the LaB6 now has a bad } coating of what I think is epoxy residue (visible under a } binoc). Does anyone have a suggestion on how to clean this safely?
Are you referring to a residue on the Wehnelt or emitter? I have seen non-conductive residue deposited onto the Wehnelt by a LaB6 (as if LaB6 itself). It was relatively difficult to remove, but a weak acid solution helped (e.g., 10% HCl).
Genuinely, Michael Shaffer :o)
SEM/MLA Research Coordinator http://www.mun.ca/creait/maf/ Inco Innovation Centre Memorial University St. John's, NL
==============================Original Headers============================== 6, 20 -- From michael-at-shaffer.net Fri Dec 15 04:50:03 2006 6, 20 -- Received: from n034.sc1.cp.net (smtpout1448.sc1.he.tucows.com [64.97.157.148]) 6, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBFAo3jk029373 6, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 15 Dec 2006 04:50:03 -0600 6, 20 -- Received: from rarewolf (205.251.83.78) by n034.sc1.cp.net (7.2.069.1) (authenticated as michael-at-shaffer.net) 6, 20 -- id 457F403C00089034; Fri, 15 Dec 2006 10:49:56 +0000 6, 20 -- From: "michael shaffer" {michael-at-shaffer.net} 6, 20 -- To: {tomw-at-uidaho.edu} 6, 20 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 6, 20 -- Subject: RE: [Microscopy] viaWWW: Cleaning a Lab6 emitter 6, 20 -- Date: Fri, 15 Dec 2006 07:21:57 -0330 6, 20 -- Message-ID: {005301c72037$0b820930$4701a8c0-at-rarewolf} 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 11 6, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 6, 20 -- Thread-Index: Accf3W9d/0RRYqWPQwG+blCThWXrrAAWP7oA 6, 20 -- In-Reply-To: {200612150012.kBF0C8ZZ006213-at-ns.microscopy.com} ==============================End of - Headers==============================
Hello, I would not suggest to used 10% HCl for cleaning Whenelt assembly. 10% HCl solution might be too aggressive for highly polished stainless steel surface. In our lab, we routinely use ammonia solution. Is it also possible to use alcoholic solution of KOH. However, you can clean the stainless parts of Wehnelt, only. All brass components should be dismounted prior cleaning in ammonia or KOH solutions. Please, see the users guide for proper cleaning of your Wehnelt. If you would like, I can make a copy of cleaning procedure of Whenelt assembly from Philips CM100 users guide manual.
Best regards Oldrich
------------------------------------------ Oldrich Benada Institute of Microbiology, Acad. Sci. CR Videnska 1083 142 20 Praha 4 - Krc Czech Republic
On 15 Dec 2006 at 4:52, michael-at-shaffer.net wrote:
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------------------------------------------ Oldrich Benada Mikrobiologický ústav AV CR Videnska 1083 142 20 Praha 4 - Krc
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Hello everyone, I routine clean my stainless steel Wehnelt with a little dilute NH3OH, but I'm removing tungsten, I'm not sure what you're clean off with a LaB6 filament.
But I would never use NH3OH or NaOH to clean brass parts. Most commercial brass and copper cleaners proudly exclaim "Does not contain ammonia." There is a reason for that. I sometime use an ammonia solution to strip copper fouling off steel, but I;m interested in cleaning the steel and not preserving the copper. I'm not saying you're wrong, but I would be very hesitant to clean brass in a basic solution.
Stay safe........ Frank Karl
Degussa Corporation
Akron Technical Center
3500 Embassy Parkway
Suite 100
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SEM/EDS has been important analytical instrumentation for the FBI for decades. We have recently expanded its utility in investigative roles, however, and therefore would like to ask the community for assistance. Whereas most SEM/EDS users need answers provided by quantitation and structural characterization, forensic inquiries generally necessitate “identifications†of questioned materials. This is generally possible only if the analyst is able to search his analytical results (spectra, primarily) against data from reference materials – as is the case with most other mature spectroscopies. We therefore have developed tools to archive and query the spectra and associated metadata of reference materials, and have produced a data set of thousands of materials in a variety of materials categories. Spectral and data queries against this database are consistently providing significant investigative direction. Of course quantitative analysis and other analytical techniques are additionally used where appropriate. Because the FBI has limited analytical capability and limited access to specific materials, I am appealing to you for either reference spectra that can be uploaded directly into our database, or materials that we can borrow and analyze at our facility (on a very limited, very specific basis). Literally all materials are viable candidates for inclusion – from commercial products to biologicals. Spectra must be exportable in EMSA format. Any data contributed will be restricted to FBI use only. If you would be willing to consider participating in this project, please contact me directly for details. We sincerely appreciate your consideration of this appeal!
Dennis Ward, FBI
Dennis Ward FBI Laboratory Chemistry Unit 2501 Investigation Parkway Quantico, VA 22135 v 703.632.7424
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The gun for the 1830 and associated components are compatible with Pol for cleaning and polishing. I used it for years on the gun, final apertures holder, anode aperture holder, etc.
Not sure about the LaB6 cathode however.
gary g.
At 05:27 AM 12/15/2006, you wrote:
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JEOL use their systems regulary in the UK for TEM and FE-SEM installations. In simple cases, we install ourselves, for more complex situations with multiple microscopes and changing fields, we get them to come in and design an appropriate set of coils.
Cancellation is not instantaneous - can take 1-2 seconds. So, if fields are fairly stable and only change occassionally/slowly, it can probably be cancelled, always depending on how sensitive the TEM is. If you have rapidly changing fields, then it gets more difficult and tends to be less sucessful.
Best regards, -- Larry Stoter JEOL (UK) Ltd tel: +44-(0)1707-377117, fax: +44-(0)1707-373254, e-mail: larrys-at-jeoluk.com
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==============================Original Headers============================== 6, 18 -- From larry-at-celtic.freewire.co.uk Fri Dec 15 11:40:08 2006 6, 18 -- Received: from get.freewire.net (get.freewire.net [86.54.106.202] (may be forged)) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBFHe8ST028543 6, 18 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 15 Dec 2006 11:40:08 -0600 6, 18 -- Received: from [194.164.78.187] (modem-rack4-modem187.netkonect.net [194.164.78.187]) 6, 18 -- by get.freewire.net (8.13.6/8.11.6) with ESMTP id kBFHe8mZ014452; 6, 18 -- Fri, 15 Dec 2006 17:40:09 GMT 6, 18 -- Mime-Version: 1.0 6, 18 -- Message-Id: {p06210200c1a88b415e82-at-[194.164.78.235]} 6, 18 -- In-Reply-To: {200612150815.kBF8FnX9025522-at-ns.microscopy.com} 6, 18 -- References: {200612150815.kBF8FnX9025522-at-ns.microscopy.com} 6, 18 -- Date: Fri, 15 Dec 2006 17:32:51 +0000 6, 18 -- To: jacques.faerber-at-ipcms.u-strasbg.fr, Microscopy-at-MSA.Microscopy.Com 6, 18 -- From: Larry Stoter {larry-at-celtic.freewire.co.uk} 6, 18 -- Subject: Re: [Microscopy] power line and stray field shielding 6, 18 -- Content-Type: text/plain; charset="iso-8859-1" ; format="flowed" 6, 18 -- Content-Transfer-Encoding: 8bit 6, 18 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBFHe8ST028543 ==============================End of - Headers==============================
Tenure-track faculty vacancy in the Department of Materials Science and Engineering at Lehigh University.
The department is searching for an outstanding individual who works at the interface between Materials Science and Engineering and the disciplines of bioengineering or biology .
Details may be found at: http://www.lehigh.edu/~inmatsci/research/bio_search.html
For questions regarding the application process, contact the search coordinator: Sharon Coe at slc6-at-lehigh.edu.
........... 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============================== 8, 21 -- From jae5-at-lehigh.edu Fri Dec 15 13:57:40 2006 8, 21 -- Received: from rain.CC.Lehigh.EDU (rain.CC.Lehigh.EDU [128.180.39.20]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBFJvdZF010748 8, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 15 Dec 2006 13:57:40 -0600 8, 21 -- Received: from [127.0.0.1] (Dyn055141.mat.Lehigh.EDU [128.180.55.141]) 8, 21 -- (authenticated bits=0) 8, 21 -- by rain.CC.Lehigh.EDU (8.13.8/8.13.8) with ESMTP id kBFJvX6I032405 8, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 8, 21 -- for {Microscopy-at-microscopy.com} ; Fri, 15 Dec 2006 14:57:38 -0500 8, 21 -- Message-ID: {4582FE2D.3030804-at-lehigh.edu} 8, 21 -- Date: Fri, 15 Dec 2006 14:57:33 -0500 8, 21 -- From: Alwyn Eades {jae5-at-lehigh.edu} 8, 21 -- Organization: Lehigh University 8, 21 -- User-Agent: Thunderbird 1.5.0.8 (Windows/20061025) 8, 21 -- MIME-Version: 1.0 8, 21 -- To: "MicroscopyListserver (E-mail)" {Microscopy-at-microscopy.com} 8, 21 -- Subject: Faculty Vacancy in Biomaterials 8, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 8, 21 -- Content-Transfer-Encoding: 7bit 8, 21 -- X-Virus-Scanned: ClamAV version 0.88.7, clamav-milter version 0.88.7 on rain.CC.Lehigh.EDU 8, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
There are several suggestions for cleaning the interior parts of electron microscopes in Section 2.10.4c (pp. 71-74) of my book Vacuum Methods in Electron Microscopy (available from SPI Supplies, Ladd, M. E, Taylor, etc.) that might be useful in solving the present problem. Specifically, mentioned there is a recommendation from Peter B. Sewell of LAB-6 Inc. for removing LaB6 deposits by soaking for about a minute in a solution consisting of one part concentrated hydrochloric acid and 4 parts water.
Good luck, and Happy Holidays 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:734-975-0858 Address mail to: 2911 Whittier Court Ann Arbor, MI 48104-6731
==============================Original Headers============================== 2, 14 -- From bigelow-at-engin.umich.edu Fri Dec 15 14:58:47 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 kBFKwkVA023089 2, 14 -- for {microscopy-at-microscopy.com} ; Fri, 15 Dec 2006 14:58:47 -0600 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 kBFKwi1c004942 2, 14 -- for {microscopy-at-microscopy.com} ; Fri, 15 Dec 2006 15:58:44 -0500 (EST) 2, 14 -- Mime-Version: 1.0 2, 14 -- Message-Id: {p06210203c1a8ba741ae0-at-[141.212.131.221]} 2, 14 -- Date: Fri, 15 Dec 2006 15:58:43 -0500 2, 14 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 2, 14 -- From: Wil Bigelow {bigelow-at-engin.umich.edu} 2, 14 -- Subject: [Microscopy]RE: Cleaning LaB6 guns 2, 14 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
50 mG tells me that you have a mis-wiring issue or a grounding of the neutral, or an unintentional grounding to metal to begin with. You should do an EMF survey (grid style) then hunt down the wiring problems. This should drop it to under 5 mG, likely under 3 mG.
Tony
...................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 off (317) 328-9594 fax (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%(SM)
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-----Original Message----- X-from: jacques.faerber-at-ipcms.u-strasbg.fr [mailto:jacques.faerber-at-ipcms.u-strasbg.fr] Sent: Friday, December 15, 2006 3:15 AM To: ph2-at-sprynet.com
Hi all
As we'll cecome next year a new TEM, we are studying the question to move all our microscopes in an other part of the building. But, if the place seems to have interesting side, obviously something must be wrong ! There is a power line which runs in a technical corridor in the underground, carrying some 800-1000 Amps for the supply of the whole building, and generating something like 50 mG in one of the possible room.
So, I have three questions :
First, is it possible to shield such a power line, to lower the field at it source (1" thick aluminum or cooper, or so) ? Is such a shielding technically possible, and not too expensive ? I think it would be better to try first to limit the perturbation at it source !
Secondly, what are the feedback from people working with dynamic magnetic field compensation ? How much do you have without it, and how does it work, with such a big field. Does it react fast enough, when the stray field changes.
Thirdly, a question for europeen lister, I'm looking for europeen manufactuerer of such dynamic compensation and/or static shielding. I know some, but I want to have several contacts.
Thanks in advance for help.
-- 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
} First, is it possible to shield such a power line, to lower the } field at it source (1" thick aluminum or cooper, or so) ? Is such a } shielding technically possible, and not too expensive ? I think it would } be better to try first to limit the perturbation at it source ! }
Hi Larry,
I can only address the shielding... You can employ shielding if physically practical, but do not use Al or Cu. The conductors must be totally enclosed for best result. Annealed iron/steel is good. It does not have to be extremely thick. Best is a material called Mu-metal (a hydrogen annealed Ni/Fe alloy if memory serves. Mu-metal is a bit pricy, however, and for best results work-hardening during fabrication should be minimized.
Regards, Woody White BWXT Services
==============================Original Headers============================== 7, 27 -- From nwwhite-at-bwxt.com Mon Dec 18 07:07:25 2006 7, 27 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBID7Puk004511 7, 27 -- for {microscopy-at-msa.microscopy.com} ; Mon, 18 Dec 2006 07:07:25 -0600 7, 27 -- Received: from ([131.184.13.224]) 7, 27 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.3265256; 7, 27 -- Mon, 18 Dec 2006 08:07:03 -0500 7, 27 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 7, 27 -- Mon, 18 Dec 2006 08:07:03 -0500 7, 27 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 7, 27 -- Content-class: urn:content-classes:message 7, 27 -- MIME-Version: 1.0 7, 27 -- Content-Type: text/plain; 7, 27 -- charset="us-ascii" 7, 27 -- Subject: RE: [Microscopy] Re: power line and stray field shielding 7, 27 -- Date: Mon, 18 Dec 2006 08:07:03 -0500 7, 27 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D0EF-at-BWXSPO01.BWXS.BWXTECH.NET} 7, 27 -- X-MS-Has-Attach: 7, 27 -- X-MS-TNEF-Correlator: 7, 27 -- Thread-Topic: [Microscopy] Re: power line and stray field shielding 7, 27 -- Thread-Index: AccgcCTK/AfNKel3R3StVZENGPeGGwCNFoyA 7, 27 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 7, 27 -- To: {larry-at-celtic.freewire.co.uk} , 7, 27 -- "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 7, 27 -- X-OriginalArrivalTime: 18 Dec 2006 13:07:03.0682 (UTC) FILETIME=[69698620:01C722A5] 7, 27 -- Content-Transfer-Encoding: 8bit 7, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBID7Puk004511 ==============================End of - Headers==============================
I am hoping that someone could provide schematics for a Hitachi S-530 SEM, particularly for the vacuum controller. I'm helping a college revive an old instrument and don't have the documents for it. Presently stuck on the vacuum controller not completing a pumpdown sequence.
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street Saint Charles, Illinois 60174 phone (630) 513-7093 fax (630) 513-7092 email mailto:ars-at-sem.com web www.sem.com
==============================Original Headers============================== 5, 20 -- From ars-at-sem.com Tue Dec 19 03:11:00 2006 5, 20 -- Received: from mail.sem.com (ns2.sem.com [66.167.158.194]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJ9B05f013031 5, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 19 Dec 2006 03:11:00 -0600 5, 20 -- Received: from VaioD (mars.sem.com [66.73.253.98]) 5, 20 -- by mail.sem.com (8.13.1/8.12.6) with SMTP id kBJ9PPKF042442 5, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 19 Dec 2006 03:25:25 -0600 (CST) 5, 20 -- (envelope-from ars-at-sem.com) 5, 20 -- Received: by localhost with Microsoft MAPI; Tue, 19 Dec 2006 03:11:22 -0600 5, 20 -- Message-ID: {01C7231B.5C779B20.ars-at-sem.com} 5, 20 -- From: "Allen R. Sampson" {ars-at-sem.com} 5, 20 -- Reply-To: "ars-at-sem.com" {ars-at-sem.com} 5, 20 -- To: "'Microscopy-at-microscopy.com'" {Microscopy-at-microscopy.com} 5, 20 -- Subject: SEM seeking Hitachi S-530 schematics 5, 20 -- Date: Tue, 19 Dec 2006 03:11:21 -0600 5, 20 -- Organization: Advanced Research Systems 5, 20 -- X-Mailer: Microsoft Internet E-mail/MAPI - 8.0.0.4211 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; charset="us-ascii" 5, 20 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I use Adobe Photoshop CS (v9) under Windows 2000. Yesterday, I encountered a problem when saving (as) files. Normally I selected the file type to save as, then typed in only the filename. PS would add the selected file extension (typically .TIF) as a default.
Now, if I do that, PS will save the file fine, but adds no file extention (e.g. .TIF). I have restarted the PC as well as reinstalled PS (installer-repair) and the problem persists. I tried selecting .JPG as the file type and the same symptom presented. ...No .JPG extension, only the filename. This is not a "viewing" problem. The file types are not hidden. Also, if I edit the filename and add .TIF to a previously saved file, all is well.
I have tried Adobe support to no avail. Their suggestion was to manually type in filename.TIF. This does work as long as what you type matches the actual file type, but I would feel much better to understand what is happening and make it right.
Suggestions?
Thanks, Woody White BWXT Services
==============================Original Headers============================== 8, 26 -- From nwwhite-at-bwxt.com Tue Dec 19 10:17:04 2006 8, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 8, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJGH3wa002967 8, 26 -- for {microscopy-at-msa.microscopy.com} ; Tue, 19 Dec 2006 10:17:03 -0600 8, 26 -- Received: from ([131.184.13.224]) 8, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.3299847; 8, 26 -- Tue, 19 Dec 2006 11:16:49 -0500 8, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 8, 26 -- Tue, 19 Dec 2006 11:16:50 -0500 8, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 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: Photoshop glitch - Opinions? 8, 26 -- Date: Tue, 19 Dec 2006 11:16:49 -0500 8, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D0F3-at-BWXSPO01.BWXS.BWXTECH.NET} 8, 26 -- X-MS-Has-Attach: 8, 26 -- X-MS-TNEF-Correlator: 8, 26 -- Thread-Topic: Photoshop glitch - Opinions? 8, 26 -- Thread-Index: AccjiRaz9NHYiUO/QLKut+jTCbxnPA== 8, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 8, 26 -- To: "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 8, 26 -- X-OriginalArrivalTime: 19 Dec 2006 16:16:50.0229 (UTC) FILETIME=[16BC7650:01C72389] 8, 26 -- Content-Transfer-Encoding: 8bit 8, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBJGH3wa002967 ==============================End of - Headers==============================
The problem may be spyware. Try running anti-spyware programs like Spybot Search and Destroy and also LavaSoft Ad-Aware SE. Both programs are free. No single program seems to find all spyware/adware, so by running a couple one will catch what the other misses. If that doesn't solve the issue, then read on.
Software that is installed on a computer will develop errors over time where a zero stored in memory is replaced by a one or visa versa. It may happen when a cosmic ray particle strikes a bit thereby changing it's value or by some other random process. Over time a number of these errors will accumulate and glitches develop. This is sometimes called "bit rot". The best solution is to back up all your data, format the hard drive, and perform a fresh installation of Windows (or OSx for Mac). Then continue installing Photoshop and any other programs you use. Bit rot happens to all programs. The bigger and more complex ones have greater opportunity for errors to occur and so are more likely to develop glitches. Some computer gurus recommend a fresh installation of Windows at six month or one year intervals. Bit rot is another good reason for frequent data backups too.
Take care,
Bob Carter 2000 Bayshore Road Lopez Island, WA 98261-8595
Original Message ---------------------------------------------------------------------------- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
Hello All,
I use Adobe Photoshop CS (v9) under Windows 2000. Yesterday, I encountered a problem when saving (as) files. Normally I selected the file type to save as, then typed in only the filename. PS would add the selected file extension (typically .TIF) as a default.
Now, if I do that, PS will save the file fine, but adds no file extention (e.g. .TIF). I have restarted the PC as well as reinstalled PS (installer-repair) and the problem persists. I tried selecting .JPG as the file type and the same symptom presented. ...No .JPG extension, only the filename. This is not a "viewing" problem. The file types are not hidden. Also, if I edit the filename and add .TIF to a previously saved file, all is well.
I have tried Adobe support to no avail. Their suggestion was to manually type in filename.TIF. This does work as long as what you type matches the actual file type, but I would feel much better to understand what is happening and make it right.
Suggestions?
Thanks, Woody White BWXT Services
==============================Original Headers============================== 8, 26 -- From nwwhite-at-bwxt.com Tue Dec 19 10:17:04 2006 8, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 8, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJGH3wa002967 8, 26 -- for {microscopy-at-msa.microscopy.com} ; Tue, 19 Dec 2006 10:17:03 -0600 8, 26 -- Received: from ([131.184.13.224]) 8, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.3299847; 8, 26 -- Tue, 19 Dec 2006 11:16:49 -0500 8, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 8, 26 -- Tue, 19 Dec 2006 11:16:50 -0500 8, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 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: Photoshop glitch - Opinions? 8, 26 -- Date: Tue, 19 Dec 2006 11:16:49 -0500 8, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D0F3-at-BWXSPO01.BWXS.BWXTECH.NET} 8, 26 -- X-MS-Has-Attach: 8, 26 -- X-MS-TNEF-Correlator: 8, 26 -- Thread-Topic: Photoshop glitch - Opinions? 8, 26 -- Thread-Index: AccjiRaz9NHYiUO/QLKut+jTCbxnPA== 8, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 8, 26 -- To: "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 8, 26 -- X-OriginalArrivalTime: 19 Dec 2006 16:16:50.0229 (UTC) FILETIME=[16BC7650:01C72389] 8, 26 -- Content-Transfer-Encoding: 8bit 8, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBJGH3wa002967 ==============================End of - Headers==============================
-- No virus found in this incoming message. Checked by AVG Free Edition. Version: 7.5.432 / Virus Database: 268.15.24/592 - Release Date: 12/18/2006 1:45 PM
==============================Original Headers============================== 20, 32 -- From bob-at-rockisland.com Tue Dec 19 11:41:09 2006 20, 32 -- Received: from mars.rockisland.com (dnscache2.rockisland.com [64.119.0.12]) 20, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJHf9bW016186 20, 32 -- for {Microscopy-at-microscopy.com} ; Tue, 19 Dec 2006 11:41:09 -0600 20, 32 -- Received: from localhost (localhost [127.0.0.1]) 20, 32 -- by localhost.rockisland.com (Postfix) with ESMTP id 0080D5976; 20, 32 -- Tue, 19 Dec 2006 09:41:08 -0800 (PST) 20, 32 -- (envelope-from bob-at-rockisland.com) 20, 32 -- X-Virus-Scanned: amavisd-new at rockisland.com 20, 32 -- Received: from mars.rockisland.com ([127.0.0.1]) 20, 32 -- by localhost (smtp.rockisland.com [127.0.0.1]) (amavisd-new, port 10024) 20, 32 -- with ESMTP id YFguJksJEHPG; Tue, 19 Dec 2006 09:41:08 -0800 (PST) 20, 32 -- Received: from housecat (adsl-sj-11-138.rockisland.net [64.119.11.138]) 20, 32 -- by mars.rockisland.com (Postfix) with SMTP id 6746E589B; 20, 32 -- Tue, 19 Dec 2006 09:41:08 -0800 (PST) 20, 32 -- (envelope-from bob-at-rockisland.com) 20, 32 -- Message-ID: {001001c72394$de2a8910$0302a8c0-at-housecat} 20, 32 -- From: "Bob Carter" {bob-at-rockisland.com} 20, 32 -- To: {Microscopy-at-microscopy.com} 20, 32 -- Cc: {NWWhite-at-bwxt.com} 20, 32 -- Subject: Photoshop glitch 20, 32 -- Date: Tue, 19 Dec 2006 09:41:08 -0800 20, 32 -- MIME-Version: 1.0 20, 32 -- Content-Type: text/plain; 20, 32 -- format=flowed; 20, 32 -- charset="iso-8859-1"; 20, 32 -- reply-type=original 20, 32 -- Content-Transfer-Encoding: 7bit 20, 32 -- X-Priority: 3 20, 32 -- X-MSMail-Priority: Normal 20, 32 -- X-Mailer: Microsoft Outlook Express 6.00.2900.3028 20, 32 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 ==============================End of - Headers==============================
} I use Adobe Photoshop CS (v9) under Windows 2000. Yesterday, } I encountered a problem when saving (as) files. Normally I } selected the file type to save as, then typed in only the } filename. PS would add the selected file extension } (typically .TIF) as a default. } } Now, if I do that, PS will save the file fine, but adds no } file extention (e.g. .TIF). ...
Always your best bet for Photoshop issues is the sheer number of users who habit the Adobe forums ...
http://www.adobeforums.com
Registration is required, but it's a worthwhile resource of information.
cheerios :o) michael shaffer
SEM/MLA Research Coordinator {http://www.mun.ca/creait/maf/} Inco Innovation Centre Memorial University St. John's, Newfoundland
==============================Original Headers============================== 9, 20 -- From michael-at-shaffer.net Tue Dec 19 11:53:41 2006 9, 20 -- Received: from n034.sc1.cp.net (smtpout1471.sc1.he.tucows.com [64.97.157.171]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJHrfCZ027344 9, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 19 Dec 2006 11:53:41 -0600 9, 20 -- Received: from roamingwolf (134.153.130.141) by n034.sc1.cp.net (7.2.069.1) (authenticated as Michael-at-Shaffer.net) 9, 20 -- id 4586853D00045DF2; Tue, 19 Dec 2006 17:52:26 +0000 9, 20 -- From: "michael shaffer" {michael-at-shaffer.net} 9, 20 -- To: {NWWhite-at-bwxt.com} 9, 20 -- Cc: "MSA Microscopy list" {Microscopy-at-microscopy.com} 9, 20 -- Subject: RE: [Microscopy] Photoshop glitch - Opinions? 9, 20 -- Date: Tue, 19 Dec 2006 14:22:24 -0330 9, 20 -- Message-ID: {001601c72396$71b3e4f0$8d829986-at-roamingwolf} 9, 20 -- MIME-Version: 1.0 9, 20 -- Content-Type: text/plain; 9, 20 -- charset="us-ascii" 9, 20 -- Content-Transfer-Encoding: 7bit 9, 20 -- X-Mailer: Microsoft Office Outlook 11 9, 20 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.3028 9, 20 -- Thread-Index: AccjiRYApncSYdNFSACOgwf3TaHIIwADNkog 9, 20 -- In-Reply-To: {200612191617.kBJGHw1t003710-at-ns.microscopy.com} ==============================End of - Headers==============================
Thanks for all the suggestions. When I have the chance, I will completely remove PS entirely and re-install fresh instead of an installer "repair". This is a pain since I have 3 upgrades - Have to start with 5.0 and work my way up :(
However, no fix yet. Was hoping the preference box had somehow un-checked itself. But in looking, I find the only choice given is upper or lower case, not whether or not to use an extension. ...Unless that choice is accidentally missing - see below.
Jim... I feel you pain! Several times when I have gone to edit} preferences} file handling, the window pops up with all the buttons there - but no text defining them! ...Resembles your problem. If I go to another preference, then return to the file window, the buttons *and* text are then displayed. Weird!
Will try the user group Michael suggested ASAP, but with the holidays approaching, may have to back-burner the problem and meet some customer deadlines. ...I am getting samples by the wheelbarrow load ;)
Hope you all have an enjoyable holiday.
Woody
==============================Original Headers============================== 9, 26 -- From nwwhite-at-bwxt.com Tue Dec 19 12:17:08 2006 9, 26 -- Received: from bwxt.com (lynsmtp1.bwxt.com [131.184.146.8]) 9, 26 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJIH8dR006212 9, 26 -- for {microscopy-at-msa.microscopy.com} ; Tue, 19 Dec 2006 12:17:08 -0600 9, 26 -- Received: from ([131.184.13.224]) 9, 26 -- by lynsmtp1.bwxt.com with ESMTP id 5202488.3301690; 9, 26 -- Tue, 19 Dec 2006 13:16:45 -0500 9, 26 -- Received: from BWXSPO01.BWXS.BWXTECH.NET ([131.184.13.31]) by bwxslynbh01.BWXS.BWXTECH.NET with Microsoft SMTPSVC(6.0.3790.1830); 9, 26 -- Tue, 19 Dec 2006 13:16:45 -0500 9, 26 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 9, 26 -- Content-class: urn:content-classes:message 9, 26 -- MIME-Version: 1.0 9, 26 -- Content-Type: text/plain; 9, 26 -- charset="us-ascii" 9, 26 -- Subject: RE: [Microscopy] Photoshop glitch - Opinions? 9, 26 -- Date: Tue, 19 Dec 2006 13:16:44 -0500 9, 26 -- Message-ID: {360DDAABED436B49BA397357CCEE8BB703D0F4-at-BWXSPO01.BWXS.BWXTECH.NET} 9, 26 -- X-MS-Has-Attach: 9, 26 -- X-MS-TNEF-Correlator: 9, 26 -- Thread-Topic: [Microscopy] Photoshop glitch - Opinions? 9, 26 -- Thread-Index: Accji7D8AEEDX6wsR52QuX2QpZURFgACvHmw 9, 26 -- From: "White, Woody N." {NWWhite-at-bwxt.com} 9, 26 -- To: "MicroscopyListServer" {Microscopy-at-msa.microscopy.com} 9, 26 -- X-OriginalArrivalTime: 19 Dec 2006 18:16:45.0355 (UTC) FILETIME=[D75D8FB0:01C72399] 9, 26 -- Content-Transfer-Encoding: 8bit 9, 26 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBJIH8dR006212 ==============================End of - Headers==============================
Many (if not most) computers today employ ECC or other types of parity checking memory. In these cases, bit level errors in memory are automatically corrected for.
I also have to say that while I've certainly experienced problems I could trace to malware or hard drive failures, I've never see never anything that I could attribute to so called "bit rot". john
At 09:47 AM 12/19/2006, you wrote: } Software that is installed on a computer will develop errors over time } where a zero stored in memory is replaced by a one or visa versa. It may } happen when a cosmic ray particle strikes a bit thereby changing it's value } or by some other random process. Over time a number of these errors will } accumulate and glitches develop. This is sometimes called "bit rot". The } best solution is to back up all your data, format the hard drive, and } perform a fresh installation of Windows (or OSx for Mac). Then continue } installing Photoshop and any other programs you use. Bit rot happens to all } programs. The bigger and more complex ones have greater opportunity for } errors to occur and so are more likely to develop glitches. Some computer } gurus recommend a fresh installation of Windows at six month or one year } intervals. Bit rot is another good reason for frequent data backups too. } } Take care, } } Bob Carter } 2000 Bayshore Road } Lopez Island, WA 98261-8595
John J. Donovan donovan-at-uoregon.edu University of Oregon (541) 346-4632 (office) 1260 Franklin Blvd (541) 346-4655 (probe) Eugene, OR (541) 346-4778 (SEM) 97403-1272 (541) 346-4692 (FAX)
Lab Web: http://epmalab.uoregon.edu/
==============================Original Headers============================== 6, 21 -- From donovan-at-uoregon.edu Tue Dec 19 13:39:07 2006 6, 21 -- Received: from smtp.uoregon.edu (mserv4.uoregon.edu [128.223.142.54]) 6, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJJd7BX019221 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 19 Dec 2006 13:39:07 -0600 6, 21 -- Received: from Probev.uoregon.edu (probev.uoregon.edu [128.223.92.75]) 6, 21 -- (authenticated bits=0) 6, 21 -- by smtp.uoregon.edu (8.13.7/8.13.7) with ESMTP id kBJJd6GB015528 6, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NOT) 6, 21 -- for {microscopy-at-microscopy.com} ; Tue, 19 Dec 2006 11:39:06 -0800 6, 21 -- Message-Id: {7.0.1.0.0.20061219113100.03a03d20-at-uoregon.edu} 6, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 6, 21 -- Date: Tue, 19 Dec 2006 11:36:31 -0800 6, 21 -- To: microscopy-at-microscopy.com 6, 21 -- From: John Donovan {donovan-at-uoregon.edu} 6, 21 -- Subject: Re: [Microscopy] Photoshop glitch 6, 21 -- In-Reply-To: {200612191747.kBJHl1Tv025837-at-ns.microscopy.com} 6, 21 -- References: {200612191747.kBJHl1Tv025837-at-ns.microscopy.com} 6, 21 -- Mime-Version: 1.0 6, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 6, 21 -- X-Virus-Scanned: ClamAV 0.88.7/2358/Tue Dec 19 08:15:46 2006 on mserv4 6, 21 -- X-Virus-Status: Clean ==============================End of - Headers==============================
Photoshop 9 aka CS2 DOES have a preference for whether or not to add the extension. In the file-handling preferences it is Append File Extension: Always/Never/Ask. Probably what has happened is simply that your preferences file has gotten corrupted. So throw it into the trash and let Photoshop create a new one when you next run it. Chances are that will fix your problem.
On Dec 19, 2006, at 1:17 PM, NWWhite-at-bwxt.com wrote:
} However, no fix yet. Was hoping the preference box had somehow } un-checked itself. But in looking, I find the only choice given is } upper } or lower case, not whether or not to use an extension. ...Unless that } choice is accidentally missing - see below.
John Russ
==============================Original Headers============================== 8, 21 -- From DrJohnRuss-at-aol.com Tue Dec 19 13:45:22 2006 8, 21 -- Received: from imo-m22.mail.aol.com (imo-m22.mx.aol.com [64.12.137.3]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJJjMDY028450 8, 21 -- for {microscopy-at-msa.microscopy.com} ; Tue, 19 Dec 2006 13:45:22 -0600 8, 21 -- Received: from DrJohnRuss-at-aol.com 8, 21 -- by imo-m22.mx.aol.com (mail_out_v38_r7.6.) id 3.d0b.474f939 (49383); 8, 21 -- Tue, 19 Dec 2006 14:45:17 -0500 (EST) 8, 21 -- Received: from [192.168.123.187] (cpe-065-190-140-239.nc.res.rr.com [65.190.140.239]) by ciaaol-d06.mail.aol.com (v114.2) with ESMTP id MAILCIAAOLD065-c0e74588414a38d; Tue, 19 Dec 2006 14:45:15 -0500 8, 21 -- Mime-Version: 1.0 (Apple Message framework v752.2) 8, 21 -- In-Reply-To: {200612191817.kBJIHVG4006916-at-ns.microscopy.com} 8, 21 -- References: {200612191817.kBJIHVG4006916-at-ns.microscopy.com} 8, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 8, 21 -- Message-Id: {60EFF99A-F17F-4BF6-9C37-5BDA0EF5F478-at-AOL.com} 8, 21 -- Content-Transfer-Encoding: 7bit 8, 21 -- From: John Russ {DrJohnRuss-at-aol.com} 8, 21 -- Subject: Re: [Microscopy] RE: Photoshop glitch - Opinions? 8, 21 -- Date: Tue, 19 Dec 2006 14:45:11 -0500 8, 21 -- To: NWWhite-at-bwxt.com, microscopy-at-msa.microscopy.com 8, 21 -- X-Mailer: Apple Mail (2.752.2) 8, 21 -- X-AOL-IP: 65.190.140.239 8, 21 -- X-Spam-Flag: NO ==============================End of - Headers==============================
I have the schematics for the S-520 SEM evacuating sequence. Is that what you're after?
John Brealey Queen Elizabeth Hospital EM Unit Adelaide, South Australia
Hi all.
I am hoping that someone could provide schematics for a Hitachi S-530 SEM, particularly for the vacuum controller. I'm helping a college revive an old instrument and don't have the documents for it. Presently stuck on the vacuum controller not completing a pumpdown sequence.
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street Saint Charles, Illinois 60174 phone (630) 513-7093 fax (630) 513-7092 email mailto:ars-at-sem.com web www.sem.com
==============================Original Headers============================== 13, 33 -- From john.brealey-at-imvs.sa.gov.au Tue Dec 19 17:21:31 2006 13, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 13, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBJNLTv6013816 13, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 19 Dec 2006 17:21:30 -0600 13, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 13, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id kBJNLSPb002066 13, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 20 Dec 2006 09:51:28 +1030 (CST)' 13, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 13, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id kBJNLSvW002061 13, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 20 Dec 2006 09:51:28 +1030 (CST)' 13, 33 -- Received: from localhost (mesh.imvs.sa.gov.au [10.20.98.37]) 13, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id 327C334BC0 13, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 20 Dec 2006 09:51:26 +1030 (CST) 13, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 13, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 13, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.37]) (amavisd-new, port 10024) 13, 33 -- with LMTP id 6Nezsvk6VH0k for {Microscopy-at-MSA.Microscopy.Com} ; 13, 33 -- Wed, 20 Dec 2006 09:51:19 +1030 (CST) 13, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 13, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 7642B34BBC 13, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 20 Dec 2006 09:51:11 +1030 (CST) 13, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 13, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 13, 33 -- Subject: SEM Schematics 13, 33 -- Date: Wed, 20 Dec 2006 09:51:09 +1030 13, 33 -- Message-ID: {000601c723c4$5e3fcc30$c88a140a-at-iqe36042} 13, 33 -- MIME-Version: 1.0 13, 33 -- Content-Type: text/plain; 13, 33 -- charset="us-ascii" 13, 33 -- Content-Transfer-Encoding: 7bit 13, 33 -- X-Mailer: Microsoft Office Outlook 11 13, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 13, 33 -- Thread-Index: AccjxFFN34uPeaL4Qn6oMFIuzqfcPg== ==============================End of - Headers==============================
Hi, I have a failure analysis problem looking at GaAs devices. It seems to me that the best way of looking for the failure would be to make a relatively thick (~1um) plan TEM section which includes the appropriate parts of the device structure. However I have the feeling that my 200 kV JEOL 2011 may not have enough oomph to get clear images. Does anyone know of a higher voltage TEM in the UK which I could hire for a day or two? Relatively low mag imaging would be good too since these devices are tens of microns wide.
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==============================Original Headers============================== 9, 31 -- From richard.beanland-at-bookham.com Wed Dec 20 04:10:30 2006 9, 31 -- Received: from mail78.messagelabs.com (mail78.messagelabs.com [195.245.230.131]) 9, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kBKAATOk006973 9, 31 -- for {microscopy-at-microscopy.com} ; Wed, 20 Dec 2006 04:10:29 -0600 9, 31 -- X-VirusChecked: Checked 9, 31 -- X-Env-Sender: richard.beanland-at-bookham.com 9, 31 -- X-Msg-Ref: server-7.tower-78.messagelabs.com!1166609427!47143150!1 9, 31 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- 9, 31 -- X-Originating-IP: [213.249.209.179] 9, 31 -- Received: (qmail 8429 invoked from network); 20 Dec 2006 10:10:27 -0000 9, 31 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) 9, 31 -- by server-7.tower-78.messagelabs.com with SMTP; 20 Dec 2006 10:10:27 -0000 9, 31 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); 9, 31 -- Wed, 20 Dec 2006 10:10:50 +0000 9, 31 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 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: High voltage TEM in the UK 9, 31 -- Date: Wed, 20 Dec 2006 10:10:49 -0000 9, 31 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E2CCAAD-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} 9, 31 -- X-MS-Has-Attach: 9, 31 -- X-MS-TNEF-Correlator: 9, 31 -- Thread-Topic: High voltage TEM in the UK 9, 31 -- thread-index: AcckHx9QMKNYlHg8RG6IQfpT5nHBBA== 9, 31 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} 9, 31 -- To: {microscopy-at-microscopy.com} 9, 31 -- X-OriginalArrivalTime: 20 Dec 2006 10:10:50.0183 (UTC) FILETIME=[1FF11D70:01C7241F] 9, 31 -- Content-Transfer-Encoding: 8bit 9, 31 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBKAATOk006973 ==============================End of - Headers==============================
Anyone know of someone capable of repairing an Ultratome V in the Southeastern US? Ours suddenly quit working, and I've exhausted the simple stuff. TIA Julian -- Julian P.S. Smith III Dept. of Biology Winthrop University 520 Cherry Rd. Rock Hill, SC 29733
Hi all, I want to set up a long-term in-vivo imaging experiment using Saivi715 microspheres (invitrogen). The fluorescence excitation and emission is 715/755 nm. Does anyone have experience with this or similar microparticles using confocal system? Thanks and greetings, Zsolt
************************************** Zsolt Lazar, PhD Molecular Cytology Core Facility Memorial Sloan-Kettering Cancer Center 415 East 68th Street, ZRC-1838 New York, NY 10021
==============================Original Headers============================== 3, 34 -- From lazarz-at-mskcc.org Wed Dec 20 13:10:20 2006 3, 34 -- Received: from extmail.mskcc.org (extmail.mskcc.org [140.163.0.99]) 3, 34 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBKJAKxn010899 3, 34 -- for {microscopy-at-microscopy.com} ; Wed, 20 Dec 2006 13:10:20 -0600 3, 34 -- Received: from 140.163.141.235 by extmail.mskcc.org with ESMTP (ESMTP 3, 34 -- Relay); Wed, 20 Dec 2006 14:10:15 -0500 3, 34 -- X-Server-Uuid: 74E28B0A-64D9-4563-A4EB-AECFC40E1CE3 3, 34 -- Received: from smskpexmbx3.MSKCC.ROOT.MSKCC.ORG ([140.163.141.252]) by 3, 34 -- smskpexsmtp2.MSKCC.ROOT.MSKCC.ORG with Microsoft 3, 34 -- SMTPSVC(6.0.3790.1830); Wed, 20 Dec 2006 14:10:15 -0500 3, 34 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 3, 34 -- Content-class: urn:content-classes:message 3, 34 -- MIME-Version: 1.0 3, 34 -- Subject: Saivi715 microspheres 3, 34 -- Date: Wed, 20 Dec 2006 14:10:14 -0500 3, 34 -- Message-ID: {E1675D0E71BA5844925CA7653D64193115AD90-at-SMSKPEXMBX3.MSKCC.ROOT.MSKCC.ORG} 3, 34 -- X-MS-Has-Attach: 3, 34 -- X-MS-TNEF-Correlator: 3, 34 -- Thread-Topic: Saivi715 microspheres 3, 34 -- Thread-Index: AcckanrIVwacOVBTSw2zEpQRpMafbg== 3, 34 -- From: LazarZ-at-mskcc.org 3, 34 -- To: microscopy-at-microscopy.com 3, 34 -- X-OriginalArrivalTime: 20 Dec 2006 19:10:15.0715 (UTC) 3, 34 -- FILETIME=[7B4D5F30:01C7246A] 3, 34 -- X-TMWD-Spam-Summary: TS=20061220191016; SEV=2.0.1; DFV=A2006122010; 3, 34 -- IFV=2.0.4,4.0-8; RPD=4.00.0004; ENG=IBF; 3, 34 -- RPDID=303030312E30413031303230332E34353839383933392E303033352D412D; 3, 34 -- CAT=NONE; CON=NONE 3, 34 -- X-MMS-Spam-Filter-ID: A2006122010_4.00.0004_4.0-8 3, 34 -- X-WSS-ID: 6997551D29O1381872-01-01 3, 34 -- Content-Type: text/plain; 3, 34 -- charset=iso-8859-2 3, 34 -- Content-Transfer-Encoding: 8bit 3, 34 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBKJAKxn010899 ==============================End of - Headers==============================
Hello, I am looking for the information of wedge polisher. I would appreciate If you could provide the information of T-tool or other wedge polisher. ( I already have the information on tripod).
Hiromi Konishi, Ph.D. University of Wisconsin-Madison
==============================Original Headers============================== 3, 30 -- From hikonishi-at-gmail.com Wed Dec 20 17:24:23 2006 3, 30 -- Received: from py-out-1112.google.com (py-out-1112.google.com [64.233.166.183]) 3, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBKNOM8u002335 3, 30 -- for {Microscopy-at-microscopy.com} ; Wed, 20 Dec 2006 17:24:23 -0600 3, 30 -- Received: by py-out-1112.google.com with SMTP id b50so1359238pyh 3, 30 -- for {Microscopy-at-microscopy.com} ; Wed, 20 Dec 2006 15:24:21 -0800 (PST) 3, 30 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 3, 30 -- s=beta; d=gmail.com; 3, 30 -- h=received:message-id:from:to:subject:date:mime-version:content-type:content-transfer-encoding:x-priority:x-msmail-priority:x-mailer:x-mimeole; 3, 30 -- b=fQiwynJBbLjEvx8NbYhgw0WATa5NZRVStAEOHR+Hal1IJU7j4ZVsVVnWOvpf31zWPJGcBNlCxm/Ez+XHL3zlqZ/20kt62Pismy+kXiwu3Llh35PxJp2G9t8rlnGSgmKhMKsiu/5ZdcPxnHRZI+95X7d55THWHKVSp8gLT7ZPBYk= 3, 30 -- Received: by 10.35.80.20 with SMTP id h20mr13861548pyl.1166657060945; 3, 30 -- Wed, 20 Dec 2006 15:24:20 -0800 (PST) 3, 30 -- Received: from HKONISHI ( [144.92.207.106]) 3, 30 -- by mx.google.com with ESMTP id f24sm12411396pyh.2006.12.20.15.24.20; 3, 30 -- Wed, 20 Dec 2006 15:24:20 -0800 (PST) 3, 30 -- Message-ID: {001301c7248d$fd25f3a0$6acf5c90-at-HKONISHI} 3, 30 -- From: "Hiromi Konishi" {hikonishi-at-gmail.com} 3, 30 -- To: {Microscopy-at-microscopy.com} 3, 30 -- Subject: wedge polisher 3, 30 -- Date: Wed, 20 Dec 2006 17:24:20 -0600 3, 30 -- MIME-Version: 1.0 3, 30 -- Content-Type: text/plain; 3, 30 -- format=flowed; 3, 30 -- charset="iso-2022-jp"; 3, 30 -- reply-type=original 3, 30 -- Content-Transfer-Encoding: 7bit 3, 30 -- X-Priority: 3 3, 30 -- X-MSMail-Priority: Normal 3, 30 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2180 3, 30 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
Many thanks to all who gave a advice about our tray field and shielding question. They confirmed our thought, and we are studying that question further to find a aceptable solution.
It's difficult to find "clean" rooms... and even more to convince non microcopists about the demands of our instruments !
Jacques
-- 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
Hi all, My Reichert Ultracut S is having a problem - the top fluorescent lights won't come on and the motor won't switch on - seems like a board in the controller unit must have gone out. Has anyone had the same experience and if so willing to give advice on fixing it?
Thanks in advance and Happy Holidays!
Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"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 Friends of the Marine Institute - Join Today! www.friendsofugami.org
==============================Original Headers============================== 11, 19 -- From beth-at-plantbio.uga.edu Thu Dec 21 12:29:14 2006 11, 19 -- Received: from dogwood.plantbio.uga.edu (dogwood.plantbio.uga.edu [128.192.26.2]) 11, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBLITEXx019746 11, 19 -- for {microscopy-at-microscopy.com} ; Thu, 21 Dec 2006 12:29:14 -0600 11, 19 -- Received: from [128.192.26.46] ([128.192.26.46]) 11, 19 -- (authenticated user beth-at-plantbio.uga.edu) 11, 19 -- by dogwood.plantbio.uga.edu 11, 19 -- (using TLSv1/SSLv3 with cipher AES128-SHA (128 bits)) 11, 19 -- for microscopy-at-microscopy.com; 11, 19 -- Thu, 21 Dec 2006 13:29:10 -0500 11, 19 -- Mime-Version: 1.0 (Apple Message framework v752.2) 11, 19 -- Content-Transfer-Encoding: 7bit 11, 19 -- Message-Id: {49D8A591-4D3E-4CC6-8D5E-7D5639FF4074-at-plantbio.uga.edu} 11, 19 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 11, 19 -- To: microscopy microscopy {microscopy-at-microscopy.com} 11, 19 -- From: Beth Richardson {beth-at-plantbio.uga.edu} 11, 19 -- Subject: problem with an Ultracut S 11, 19 -- Date: Thu, 21 Dec 2006 13:29:10 -0500 11, 19 -- X-Mailer: Apple Mail (2.752.2) ==============================End of - Headers==============================
I have sent the schematics to your email address as listed below.
John Brealey
Might be the same, I'd have to see them. Any chance you could scan and email them?
Allen R. Sampson, Owner Advanced Research Systems 317 North 4th. Street Saint Charles, Illinois 60174 phone (630) 513-7093 fax (630) 513-7092 email mailto:ars-at-sem.com web www.sem.com
On Tuesday, December 19, 2006 5:24 PM, john.brealey-at-imvs.sa.gov.au [SMTP:john.brealey-at-imvs.sa.gov.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 } ---------------------------------------------------------------------- } ------ } } I have the schematics for the S-520 SEM evacuating sequence. } Is that what you're after? } } John Brealey } Queen Elizabeth Hospital EM Unit } Adelaide, South Australia } } } } } } } Hi all. } } I am hoping that someone could provide schematics for a Hitachi S-530 } SEM, particularly for the vacuum controller. I'm helping a college } revive an old instrument and don't have the documents for it. } Presently stuck on the vacuum controller not completing a pumpdown sequence. } } Allen R. Sampson, Owner } Advanced Research Systems } 317 North 4th. Street } Saint Charles, Illinois 60174 } phone (630) 513-7093 fax (630) 513-7092 email mailto:ars-at-sem.com web } www.sem.com }
==============================Original Headers============================== 10, 33 -- From john.brealey-at-imvs.sa.gov.au Thu Dec 21 16:43:37 2006 10, 33 -- Received: from adl0741.systems.sa.gov.au (adl0741.systems.sa.gov.au [143.216.236.20]) 10, 33 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBLMhakH004158 10, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 21 Dec 2006 16:43:37 -0600 10, 33 -- Received: from adl0741.systems.sa.gov.au (localhost [127.0.0.1]) 10, 33 -- by adl0741.systems.sa.gov.au OUTGOING (8.12.10/8.12.10) with ESMTP id kBLMhYni027848 10, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 22 Dec 2006 09:13:34 +1030 (CST)' 10, 33 -- Received: from ablett.imvs.sa.gov.au (ablett.imvs.sa.gov.au [10.20.98.41]) 10, 33 -- by adl0741.systems.sa.gov.au INCOMING (8.12.10/8.12.10) with ESMTP id kBLMhY7X027845 10, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 22 Dec 2006 09:13:34 +1030 (CST)' 10, 33 -- Received: from localhost (mesh.imvs.sa.gov.au [10.20.98.37]) 10, 33 -- by ablett.imvs.sa.gov.au (Postfix) with ESMTP id 203DC34BFA 10, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 22 Dec 2006 09:13:34 +1030 (CST) 10, 33 -- X-Virus-Scanned: by amavisd-new at imvs.sa.gov.au 10, 33 -- Received: from ablett.imvs.sa.gov.au ([10.20.98.41]) 10, 33 -- by localhost (.imvs.sa.gov.au [10.20.98.37]) (amavisd-new, port 10024) 10, 33 -- with LMTP id oBHrYgoDgyKE for {Microscopy-at-MSA.Microscopy.Com} ; 10, 33 -- Fri, 22 Dec 2006 09:13:26 +1030 (CST) 10, 33 -- Received: from iqe36042 (iqepc200.imvs.sa.gov.au [10.20.138.200]) 10, 33 -- by ablett.imvs.sa.gov.au (Postfix) with SMTP id 30E4E34B82 10, 33 -- for {Microscopy-at-MSA.Microscopy.Com} ; Fri, 22 Dec 2006 09:13:26 +1030 (CST) 10, 33 -- From: "John Brealey" {john.brealey-at-imvs.sa.gov.au} 10, 33 -- To: "Listserver" {Microscopy-at-MSA.Microscopy.Com} 10, 33 -- Subject: Hitachi S-520 Schematics 10, 33 -- Date: Fri, 22 Dec 2006 09:13:24 +1030 10, 33 -- Message-ID: {000001c72551$6d1d3d80$c88a140a-at-iqe36042} 10, 33 -- MIME-Version: 1.0 10, 33 -- Content-Type: text/plain; 10, 33 -- charset="us-ascii" 10, 33 -- Content-Transfer-Encoding: 7bit 10, 33 -- X-Mailer: Microsoft Office Outlook 11 10, 33 -- Thread-Index: AcclUWAY1XiZQ4VBRMynH+qIOuItwA== 10, 33 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1807 ==============================End of - Headers==============================
I just wanted to say to all that I bought this recommended book and it is absolutely wonderful.
Thank you very much. What a wonderful addition to my library.
dj
On Thu, 7 Dec 2006, 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 } ---------------------------------------------------------------------------- } } This thread is following the route to a Nobel prize in Physics.. have a } read of Richard Feynman's "QED-the strange theory of light and matter", } which addresses exactly this point and is a summary of the work for } which he was awarded his gong. I haven't read it for a few years, but I } remember him starting with glass being 80% transmissive, and asking how } could individual photons 'know' whether to cross the glass or not to } give the right answer of 80%. Like most of his stuff it is very } readable and entertaining. And yes, the world is actually weirder than } you think it is.. } } Richard } } ________________________________________ } Richard Beanland } Materials Analysis } Bookham } Caswell } Towcester } Northants } NN12 8EQ } United Kingdom } Tel. +44 1327 356362 } Fax. +44 1327 356775 } http://www.bookham.com } ________________________________________ } } -----Original Message----- } X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu] } Sent: 06 December 2006 20:27 } To: Richard Beanland } Subject: [Microscopy] optical physics:Challenge } } } } } ------------------------------------------------------------------------ } ---- } 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 } ------------------------------------------------------------------------ } ---- } } Greetings, } These and other wonderful answers to the Snell law question } have all been wave-based. Is there an equivalent photon-based answer? } Presumably, if you fired one photon at a time at an oblique surface } you would get the most counts at the Snell angle; but, with single } photons, how are we to understand lines of coherent marchers or even } two feet on roller blades? Is this light being weird? } } Just wondering... } Tobias } } } } } This is a really fine answer. I would add one more analogy, since } } the question (I think) relates to the change in direction of the path } } of the light beam. In my classes, I refer to the image of a column } } of a marching band going from a concrete surface to a broken field. } } If they are exactly perpendicular to the boundary, they continue to } } go straight, although with some stumbles. If they enter at an angle, } } then those members who are on the "acute" side of the column slow } } down first. This "pulls the others along as well, bit by bit, and } } the overall direction of the march changes. } } } } Date sent: Wed, 6 Dec 2006 11:21:25 -0600 } } To: jbs-at-temple.edu } } X-from: bfoster-at-mme1.com } } Send reply to: bfoster-at-mme1.com } } Subject: [Microscopy] Re: viaWWW: optical physics } } } } } } } } } } } } } } } } } } } Hi, } } } } } } The answer is really quite simple: Snell's Law. The bending of } } } light as it crosses the boundary from one RI to another is governed } } } by the Laws of Refraction. } } } } } } The bending discussed in your question requires that several } } } conditions be met: First, that there be different RIs on each side } } } of the boundary. Second, that the light approach the boundary at } } } an angle. } } } } } } To understand what happens, it is first necessary to understand } } } that refractive index is actually a measure of the impact of } } } interaction between the electric field of light and the electric } } } field of matter. The greater the interaction, the more slowly } } } light will travel through that material and the higher the } } } refractive index. For instance: } } } RI = velocity of light in air/velocity of light in material } } } RI of air = 1, velocity of light = 300,000 km/s } } } RI of water is 1.33, velocity of light is 225,000 km/s } } } RI of immersion oil, glass, and many polymers is approximately } } } 1.5, velocity of light = 200,000 km/sec. } } } } } } As for refraction, the analogy which is often used is that of a } } } person roller skating (or in this day and age, roller blading) from } } } one surface to another. For example, from the concrete sidewalk } } } onto grass. The smooth surface of the concrete is analogous to a } } } material with low refractive index; the rougher surface of glass is } } } analogous to a material with higher RI. If the skater approaches } } } the sidewalk:grass boundary with both feet parallel, both feet will } } } slow down by the same amount and the skater will continue skating } } } in the same direction. However, if the skater approaches the } } } boundary at an angle, one foot will slow down while the other } } } remains at the original speed. That different in speed causes the } } } skater to pivot toward the material of higher RI. } } } } } } If you think of light in terms of a wave front rather than a } } } simple ray, the analogy transfers easily. If the wave approaches a } } } boundary at an angle, the edge of the front which hits the higher } } } RI first will slow down, causing the whole wave front to pivot } } } around that point. That's what happens, for example, when light } } } travels from air (RI = 1.00) into a glass coverslip (RI ~1.5) or a } } } droplet of water or a cell (RI ~1.33). You can use the optical } } } axis of the microscope as a reference. In these cases, the light } } } will bend in TOWARD the optical axis. } } } Conversely, when light emerges from a glass slide or a droplet of } } } water it will bend AWAY from the OA. Actually this is why we use } } } oil immersion: to cause those rays which would normally bend away } } } from the OA, causing a loss of both intensity and the critical } } } contribution to resolution and edge definition, to bend back toward } } } the OA, where they have an opportunity to be captured by the } } } objective and make a positive contribution to better imaging. } } } } } } All of this is explained in detail, with diagrams in Optimizing } } } Light Microscopy. If you are interested in a copy, please contact } } } Ken Piel here in the MME office for purchasing details (see below). } } } } } } 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 next } } } April. 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 or email him at kenpiel-at-mme1.com } } } } } } } } } } } } At 08:24 AM 12/6/2006, vthawfeek-at-yahoo.co.in wrote: } } } } } } } } } } } } } } } } } } } 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 vthawfeek-at-yahoo.co.in as well as the } } } MIcroscopy Listserver } } } } } ----------------------------------------------------------------------- } ---- } } } } } } } } Email: vthawfeek-at-yahoo.co.in } } } } Name: V.Thawfeek Mohammed } } } } } } } } Organization: psg tech coll. of engg. } } } } } } } } Title-Subject: [Filtered] optical physics } } } } } } } } Question: why does a light beam diverge when it strikes a medium } } } with greater refractive index than the medium ,in which it is. } } } } } } } } } ----------------------------------------------------------------------- } ---- } } } } } } -- } _ ____ __ ____ } / \ / / \ / \ \ Tobias I. Baskin } / / / / \ \ \ Biology Department } /_ / __ /__ \ \ \__ 611 N. Pleasant St. } / / / \ \ \ University of } Massachusetts } / / / \ \ \ Amherst, MA, 01003 } / / ___ / \ \__/ \ ____ } http://www.bio.umass.edu/biology/baskin/ } Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243 } } ==============================Original } Headers============================== } 6, 19 -- From baskin-at-bio.umass.edu Wed Dec 6 14:25:31 2006 } 6, 19 -- Received: from marlin.bio.umass.edu (marlin.bio.umass.edu } [128.119.55.19]) } 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id kB6KPVfD011840 } 6, 19 -- for {microscopy-at-microscopy.com} ; Wed, 6 Dec 2006 } 14:25:31 -0600 } 6, 19 -- Received: from [172.30.55.79] (eutopia [128.119.55.30]) } 6, 19 -- by marlin.bio.umass.edu (8.13.6/8.13.6) with ESMTP id } kB6KPQZB025363; } 6, 19 -- Wed, 6 Dec 2006 15:25:27 -0500 (EST) } 6, 19 -- Mime-Version: 1.0 } 6, 19 -- Message-Id: {p0623090dc19cd66dc686-at-[172.30.55.79]} } 6, 19 -- In-Reply-To: {200612062000.kB6K0oST001676-at-ns.microscopy.com} } 6, 19 -- References: {200612062000.kB6K0oST001676-at-ns.microscopy.com} } 6, 19 -- Date: Wed, 6 Dec 2006 15:25:24 -0500 } 6, 19 -- To: microscopy-at-microscopy.com } 6, 19 -- From: Tobias Baskin {baskin-at-bio.umass.edu} } 6, 19 -- Subject: optical physics:Challenge } 6, 19 -- Cc: jbs-at-temple.edu } 6, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 6, 19 -- X-Greylist: Sender succeded SMTP AUTH authentication, not } delayed by milter-greylist-1.6 (marlin.bio.umass.edu [128.119.55.19]); } Wed, 06 Dec 2006 15:25:29 -0500 (EST) } 6, 19 -- X-Scanned-By: MIMEDefang 2.54 on 128.119.55.19 } ==============================End of - } Headers============================== } } ======================================================================= } This e-mail is intended for the person it is addressed to only. 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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============================== } 15, 32 -- From richard.beanland-at-bookham.com Thu Dec 7 03:45:10 2006 } 15, 32 -- Received: from mail83.messagelabs.com (mail83.messagelabs.com [195.245.231.83]) } 15, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id kB79j90N032413 } 15, 32 -- for {microscopy-at-microscopy.com} ; Thu, 7 Dec 2006 03:45:10 -0600 } 15, 32 -- X-VirusChecked: Checked } 15, 32 -- X-Env-Sender: richard.beanland-at-bookham.com } 15, 32 -- X-Msg-Ref: server-15.tower-83.messagelabs.com!1165484708!42046564!1 } 15, 32 -- X-StarScan-Version: 5.5.10.7; banners=bookham.com,-,- } 15, 32 -- X-Originating-IP: [213.249.209.179] } 15, 32 -- Received: (qmail 31146 invoked from network); 7 Dec 2006 09:45:08 -0000 } 15, 32 -- Received: from unknown (HELO cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI) (213.249.209.179) } 15, 32 -- by server-15.tower-83.messagelabs.com with SMTP; 7 Dec 2006 09:45:08 -0000 } 15, 32 -- Received: from cas-smx-02.BOOKHAM.ENTERPRISE.PRI ([10.4.0.223]) by cas-smx-brdg-02.BOOKHAM.ENTERPRISE.PRI with Microsoft SMTPSVC(6.0.3790.1830); } 15, 32 -- Thu, 7 Dec 2006 09:45:15 +0000 } 15, 32 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 } 15, 32 -- Content-class: urn:content-classes:message } 15, 32 -- MIME-Version: 1.0 } 15, 32 -- Content-Type: text/plain; } 15, 32 -- charset="us-ascii" } 15, 32 -- Subject: RE: [Microscopy] optical physics:Challenge } 15, 32 -- Date: Thu, 7 Dec 2006 09:45:14 -0000 } 15, 32 -- Message-ID: {9645D3E33E4C6548B12A7B25F611533E2CC54D-at-cas-smx-02.BOOKHAM.ENTERPRISE.PRI} } 15, 32 -- X-MS-Has-Attach: } 15, 32 -- X-MS-TNEF-Correlator: } 15, 32 -- Thread-Topic: [Microscopy] optical physics:Challenge } 15, 32 -- thread-index: AccZdN5xFl4Td6sqTl6++eu/sEqL/AAbkU2w } 15, 32 -- From: "Richard Beanland" {richard.beanland-at-bookham.com} } 15, 32 -- To: {baskin-at-bio.umass.edu} } 15, 32 -- Cc: {microscopy-at-microscopy.com} } 15, 32 -- X-OriginalArrivalTime: 07 Dec 2006 09:45:15.0631 (UTC) FILETIME=[65E84FF0:01C719E4] } 15, 32 -- Content-Transfer-Encoding: 8bit } 15, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kB79j90N032413 } ==============================End of - Headers============================== }
==============================Original Headers============================== 6, 19 -- From dljones-at-bestweb.net Fri Dec 22 09:43:57 2006 6, 19 -- Received: from vms042pub.verizon.net (vms042pub.verizon.net [206.46.252.42]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBMFhvFu006278 6, 19 -- for {Microscopy-at-microscopy.com} ; Fri, 22 Dec 2006 09:43:57 -0600 6, 19 -- Received: from localhost ([71.249.4.219]) 6, 19 -- by vms042.mailsrvcs.net (Sun Java System Messaging Server 6.2-6.01 (built Apr 6, 19 -- 3 2006)) with ESMTPA id {0JAO00480MCXOUP7-at-vms042.mailsrvcs.net} for 6, 19 -- Microscopy-at-microscopy.com; Fri, 22 Dec 2006 09:43:50 -0600 (CST) 6, 19 -- Date: Fri, 22 Dec 2006 10:43:45 -0500 (Eastern Standard Time) 6, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 6, 19 -- Subject: Re: [Microscopy] RE: optical physics:Challenge 6, 19 -- In-reply-to: {200612070947.kB79lwSs006322-at-ns.microscopy.com} 6, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 6, 19 -- To: richard.beanland-at-bookham.com 6, 19 -- Cc: Microscopy-at-microscopy.com 6, 19 -- Message-id: {Pine.WNT.4.64.0612221037410.3156-at-H-F1} 6, 19 -- MIME-version: 1.0 6, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 6, 19 -- References: {200612070947.kB79lwSs006322-at-ns.microscopy.com} ==============================End of - Headers==============================
For those who would like to get this from the horses mouth, as it were, I discovered a link that Feynman gave at the University of Auckland in 1979. This one is about properties of light, and accounts for bending of light and lenses (see about 52 minutes into the lecture) in a way that attempts to resolve the particle/wave issue. http://www.vega.org.uk/video/programme/46
Joel
Date sent: Fri, 22 Dec 2006 09:44:08 -0600 To: jbs-at-temple.edu X-from: dljones-at-bestweb.net Send reply to: dljones-at-bestweb.net
This Question was submitted to Ask-A-Microscopist by (tauria-at-hotmail.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, December 27, 2006 at 05:19:51 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both tauria-at-hotmail.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: tauria-at-hotmail.com Name: DR. FRANCIS J. PRONESTI
Organization: WORLD ENERGY SERVICES, LTD.
Education: Graduate College
Location: CASTELLANA GROTTE, BARI, ITALY
Title: ZEISS TEM 805 DOCUMENTATION
Question: HELLO, I NEED DESPERATELY TO BUY, BORROW OR STEAL A COPY OF SETPUP, OPERATION AND MAINTENANCE FOR A CZRL ZEISS TEM MICROSCOPE, MODEL 805.
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 mark.grimson-at-ttu.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mark.grimson-at-ttu.edu Name: Mark Grimson
Organization: Texas Tech University
Title-Subject: [Filtered] 3-D reconstruction from serial paraffin sections
Question: I am attempting to follow the path of cotton fibers around a seed in an attempt to see how they pack as they develop. I have embeedded the samples in paraffin and serially sectioned through the bundle in 12 micron sections. As such, I have almost 900 sections. Is there a program or method that can generate a 3-D reconstruction from some or all of these 2-D sections via light microscopy with a digital camera? Thank you. Mark
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Question: I am looking for the information of wedge polisher. I would appreciate it if you could provide the information of T-tool or other wedge polisher. ( I already have the information on tripod).
Thank you, Hiromi Konishi, Ph.D. University of Wisconsin-Madison
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 pedromfjcosta-at-googlemail.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: pedromfjcosta-at-googlemail.com Name: Pedro MFJ Costa
Organization: National Institute of Materials Science
Title-Subject: [Filtered] TEM current density measurements
Question: Dear Listers,
As part of a series of studies I am doing on beam damage of semiconductor structures, I have recently taken some measurements of the electron current in our JEOL 300 kV microscope. For this I relied on a retractable Faraday cup (located in the viewing chamber). My approach was as follows: I decided to investigate two modes (Low Mag and Mag1). This choice is justified by the fact that these are the ones we use for the IV experiments. For each mode I chose some representative magnifications to see if the effect of changing Mag would make any difference (I was not expecting any, but I thought it would be advisable to check). Within each Magnification I took readings using the Faraday cup (current values were read from a picoammeter). Since the Faraday cup was out-centred relative to the optical axis of the microscope, I had to shift the beam in between readings so that it would be centred with the Faraday cup's entrance axis (the need for centring comes from the fact that the intensity distribution is not uniform on the illuminated area but rather likely to follow a Gaussian-like curve). Taking into consideration that the entrance hole in the Faraday cup has a diameter of 1 mm, this means that the collection area will be: A = î?.r2 = 0.007854 cm2. So, if we have, for instance, 10000 pA read from the picoammeter for a Mag = 300 in an area of 0.007854 cm2 this would directly correspond to 1.273 ??A/cm2. In my experiments I used different measures of beam spread (fully converged beam, spreaded beam ?¯ limited by the negative makrers in the vieweing chamber fluorescent screen, almost parallel beam ?¯ approximately the size of the chamber viewing screen. So, from the above, using the aforementioned conditions, we get a maximum current density of 8.913 ??A/cm2 and a minimum of 0.382 nA/cm2. If finally we consider the relation for current density, j = eNv, where j is current density (A/cm2), e is the charge of an electron (C), N is the number of electrons per unit volume (particles/cm3) and v is the velocity of the electron, then we can estimate the electron flux density (i.e. the number of electrons per unit area per unit time) and this will give us, Minimum = 2.38E109 electrons/cm2/s, Maximum = 5.56E1013 electrons/cm2/s. Now my problem is whether my reasoning is correct. This is because I have seen many studies in the literature where current density values can be much higher (order of A/cm2). True that I am using a condenser aperture and this limits the number of electrons that we get from the gun. Also I expect that the settings of the guns electrostatic lens would make a difference but this, I suppose, is not accessible to the common user to change it. So, basically, it is mostly a matter of how much the beam is converged when it hits the sample. In my case no sample was used so there are no problems as to the electrons being backscattered/Rutherford scattered/etc by the specimen. For instance, I would expect that most of the electrons that come out from the condenser aperture would ultimately be collected in the reading devices (focus screen and Faraday cup) particularly for a converged beam (naturally there may be a percentage of electrons lost on their way down the column as they backscatter through the different column components but I am assuming that these are not significant). I understand that the density of collected electrons is not going to be similar in the Faraday cup, focus screen and fluorescent screen, as all these are in different imaging planes. Ideally we would like to have a holder that seats at the same height as the sample with a Faraday cup in it and take our readings at the same conditions as the imaging experimental settings. I think nonetheless that it may still be possible to use these readings and at least show a fairly reliable interval of current densities for our measures. So my questions are: Are my calculations correct in terms procedure? How come the values I get are so different from other quoted figures?
My best wishes,
Pedro -- Pedro MFJ Costa National Institute of Materials Science 1-1 Namiki, Tsukuba Ibaraki 305-0044 Japan
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (lazarz-at-mskcc.org) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Wednesday, December 20, 2006 at 13:00:52 ---------------------------------------------------------------------------
Email: lazarz-at-mskcc.org Name: Zsolt Lazar
Organization: Memorial Sloan-Kettering Cancer Center
Education: Graduate College
Location: New York, NY
Question: Hi all, I want to set up a long-term in-vivo imaging experiment using Saivi715 microspheres (invitrogen). The fluorescence excitation and emission is 715/755 nm. Does anyone have experience with this or similar microparticles using confocal system? Thanks and greetings, Zsolt
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Organization: Comision Nacional de Energia Atomica- Argentina
Title-Subject: [Filtered] Problems with the EMS on line
Question: Dear Listers I used to run the free EMS on line (http://cimesg1.epfl.ch/CIOL) to draw SAD and Kikuchi patterns, (hkl) distances, etc., but I am having problems now. The legend "This calcul has failed for an undefined reason"always appeared when I try to run a routine. Can any body help me with it? Thanks in advanced! Patricia
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Email: onkel98-at-yahoo.com Name: pipetman
Title-Subject: [Filtered] Q: understanding STED?
Question: I have a problem conceptually understanding STED.
I can see, how a diffraction-limited spot of excited fluorophores is shaped to narrow the PSF. As a result, I get something akin to a point source of emission. But as the emitted light travels back through the microscope's optics, it is subject to diffraction again (and should thus destroy the narrowed PSF).
Is there somewhere a Gaussian fit applied at the other end (similar to PALM) to trace back the original spot? Or am I missing something basic here?
Any explanation would be greatly appreciated. Thanks - p
Check out 3D Constructor from Media Cybernetics: http://www.mediacy.com/index.aspx?page=3DConstructor
Best regards,
Walter F. Bobrowski Sr. Scientist Pfizer Global R&D 2800 Plymouth Rd. Ann Arbor, MI 48105
TEL: 734-622-7814 FAX: 734-622-5718
"The ultimate human freedom is the ability to choose one's attitude in a given set of circumstances." -Viktor Frankl
-----Original Message----- X-from: mark.grimson-at-ttu.edu [mailto:mark.grimson-at-ttu.edu] Sent: Wednesday, December 27, 2006 9:08 AM To: Bobrowski, Walter
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Email: mark.grimson-at-ttu.edu Name: Mark Grimson
Organization: Texas Tech University
Title-Subject: [Filtered] 3-D reconstruction from serial paraffin sections
Question: I am attempting to follow the path of cotton fibers around a seed in an attempt to see how they pack as they develop. I have embeedded the samples in paraffin and serially sectioned through the bundle in 12 micron sections. As such, I have almost 900 sections. Is there a program or method that can generate a 3-D reconstruction from some or all of these 2-D sections via light microscopy with a digital camera? Thank you. Mark
==============================Original Headers============================== 6, 12 -- From zaluzec-at-microscopy.com Wed Dec 27 07:59:57 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 kBRDxujB025496 6, 12 -- for {microscopy-at-microscopy.com} ; Wed, 27 Dec 2006 07:59:56 -0600 6, 12 -- Mime-Version: 1.0 6, 12 -- X-Sender: (Unverified) 6, 12 -- Message-Id: {p06110406c1b82ccd87e2-at-[206.69.208.22]} 6, 12 -- Date: Wed, 27 Dec 2006 07:59:55 -0600 6, 12 -- To: microscopy-at-microscopy.com 6, 12 -- From: mark.grimson-at-ttu.edu (by way of MicroscopyListserver) 6, 12 -- Subject: viaWWW: 3-D reconstruction from serial paraffin sections 6, 12 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers============================== ---------------------------------------------------------------------- LEGAL NOTICE Unless expressly stated otherwise, this message is confidential and may be privileged. It is intended for the addressee(s) only. Access to this E-mail by anyone else is unauthorized. If you are not an addressee, any disclosure or copying of the contents of this E-mail or any action taken (or not taken) in reliance on it is unauthorized and may be unlawful. If you are not an addressee, please inform the sender immediately.
==============================Original Headers============================== 19, 29 -- From Walter.Bobrowski-at-pfizer.com Wed Dec 27 20:33:02 2006 19, 29 -- Received: from mopmsgoa01.pfizer.com (mopmsgo.pfizer.com [148.168.100.84]) 19, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBS2X16u015093 19, 29 -- for {microscopy-at-microscopy.com} ; Wed, 27 Dec 2006 20:33:02 -0600 19, 29 -- Received: from groamrexc01.amer.pfizer.com (groamrexc01.amer.pfizer.com [172.30.8.168]) 19, 29 -- by mopmsgoa01.pfizer.com (8.13.4/8.13.4) with ESMTP id kBS2ViuL003247; 19, 29 -- Wed, 27 Dec 2006 21:31:44 -0500 19, 29 -- Received: from mopamrexc03.amer.pfizer.com ([170.116.30.69]) by groamrexc01.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 19, 29 -- Wed, 27 Dec 2006 21:33:04 -0500 19, 29 -- Received: from anaamrexm01.amer.pfizer.com ([162.48.101.10]) by mopamrexc03.amer.pfizer.com with Microsoft SMTPSVC(6.0.3790.1830); 19, 29 -- Wed, 27 Dec 2006 21:33:03 -0500 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: [Microscopy] viaWWW: 3-D reconstruction from serial paraffin sections 19, 29 -- Date: Wed, 27 Dec 2006 21:33:02 -0500 19, 29 -- Message-ID: {CF89AF6ABD93B746A55BCCD9480A4B3D0844FE99-at-anaamrexm01.amer.pfizer.com} 19, 29 -- X-MS-Has-Attach: 19, 29 -- X-MS-TNEF-Correlator: 19, 29 -- Thread-Topic: [Microscopy] viaWWW: 3-D reconstruction from serial paraffin sections 19, 29 -- Thread-Index: AccpwHkH8NeqHHlRRSyqJkKUBwET+QAZ2pfQ 19, 29 -- From: "Bobrowski, Walter" {Walter.Bobrowski-at-pfizer.com} 19, 29 -- To: {mark.grimson-at-ttu.edu} , {microscopy-at-microscopy.com} 19, 29 -- X-OriginalArrivalTime: 28 Dec 2006 02:33:03.0703 (UTC) FILETIME=[7FF2DE70:01C72A28] 19, 29 -- X-Proofpoint-Spam-Reason: safe 19, 29 -- Content-Transfer-Encoding: 8bit 19, 29 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id kBS2X16u015093 ==============================End of - Headers==============================
You might look at Reconstruct, http://synapses.bu.edu/tools/index.htm
It focused on reconstruction of serial TEM images but would probably work well for this application.
- Davi
Davi Bock Ph.D. student, Program in Neuroscience Harvard University
On Wed, Dec 27, 2006 at 08:07:03AM -0600, mark.grimson-at-ttu.edu wrote: } } Email: mark.grimson-at-ttu.edu } Name: Mark Grimson } } Organization: Texas Tech University } } Title-Subject: [Filtered] 3-D reconstruction from serial paraffin sections } } Question: I am attempting to follow the path of cotton fibers around a seed in an attempt to see how they pack as they develop. I have embeedded the samples in paraffin and serially sectioned through the bundle in 12 micron sections. As such, I have almost 900 sections. Is there a program or method that can generate a 3-D reconstruction from some or all of these 2-D sections via light microscopy with a digital camera? Thank you. Mark }
==============================Original Headers============================== 6, 17 -- From dbock-at-bostoncoop.net Thu Dec 28 11:16:50 2006 6, 17 -- Received: from bostoncoop.net (outbound.bostoncoop.net [72.1.169.236]) 6, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id kBSHGobJ009093 6, 17 -- for {microscopy-at-microscopy.com} ; Thu, 28 Dec 2006 11:16:50 -0600 6, 17 -- Received: by bostoncoop.net (Postfix, from userid 1011) 6, 17 -- id 6BA12A0146; Thu, 28 Dec 2006 12:16:49 -0500 (EST) 6, 17 -- Date: Thu, 28 Dec 2006 12:16:49 -0500 6, 17 -- From: Davi Bock {dbock-at-hms.harvard.edu} 6, 17 -- To: microscopy-at-microscopy.com 6, 17 -- Subject: Re: [Microscopy] viaWWW: 3-D reconstruction from serial paraffin sections 6, 17 -- Message-ID: {20061228171649.GA20306-at-bostoncoop.net} 6, 17 -- References: {200612271407.kBRE73GO023123-at-ns.microscopy.com} 6, 17 -- Mime-Version: 1.0 6, 17 -- Content-Type: text/plain; charset=us-ascii 6, 17 -- Content-Disposition: inline 6, 17 -- In-Reply-To: {200612271407.kBRE73GO023123-at-ns.microscopy.com} 6, 17 -- User-Agent: Mutt/1.5.9i ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (wawennekes-at-woh.rr.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, December 29, 2006 at 20:19:28 ---------------------------------------------------------------------------
Email: wawennekes-at-woh.rr.com Name: Willem Wennekes
Organization: UES
Education: Graduate College
Location: Dayton, Ohio, USA
Question: What is the difference in performance between a solid state & a scintilater (Robinson) backscatter detector? I am very interested in a side by side comparison.