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
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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.
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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.
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-- ........... 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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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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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
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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
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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
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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==============================Original Headers============================== 11, 21 -- From gary-at-gaugler.com Thu Feb 2 20:20:12 2006 11, 21 -- Received: from qsmtp2.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 11, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k132KCki016497 11, 21 -- for {microscopy-at-microscopy.com} ; Thu, 2 Feb 2006 20:20:12 -0600 11, 21 -- Received: (qmail 387 invoked from network); 2 Feb 2006 18:20:12 -0800 11, 21 -- Received: by simscan 1.1.0 ppid: 384, pid: 385, t: 0.1682s 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; 2 Feb 2006 18:20:12 -0800 11, 21 -- Message-Id: {6.2.3.4.2.20060202180159.02002198-at-mail.calweb.com} 11, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 11, 21 -- Date: Thu, 02 Feb 2006 18:20:14 -0800 11, 21 -- To: Judith_A_Ruiz-at-whirlpool.com 11, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 11, 21 -- Subject: Re: [Microscopy] viaWWW: lab service for hire, How much do you 11, 21 -- charge 11, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 11, 21 -- In-Reply-To: {200602030144.k131ioQN004044-at-ns.microscopy.com} 11, 21 -- References: {200602030144.k131ioQN004044-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 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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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.
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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
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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.
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 --
==============================Original Headers============================== 8, 25 -- From jcumings-at-gmail.com Thu Feb 9 08:35:58 2006 8, 25 -- Received: from uproxy.gmail.com (uproxy.gmail.com [66.249.92.204]) 8, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k19EZv1W027008 8, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 9 Feb 2006 08:35:58 -0600 8, 25 -- Received: by uproxy.gmail.com with SMTP id s2so260073uge 8, 25 -- for {Microscopy-at-microscopy.com} ; Thu, 09 Feb 2006 06:35:57 -0800 (PST) 8, 25 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 25 -- s=beta; d=gmail.com; 8, 25 -- h=received:message-id:date:from:reply-to:sender:to:subject:mime-version:content-type:content-transfer-encoding:content-disposition; 8, 25 -- b=J7Yh3j/yK8qwQ23qXD6S/Pwrua8D74E3rIXGt9kBx4eFw1tKTqNnPy7smG5lUD3ppuvFgteak9KXbxsNCbt+yyJ4pxWsBJTngWRcNa97f2Faq+nWy5TFTU80zgJW5XqVm16M9Jh1a1he3KQXpbV+6SwDYRIc7trYXy9XsKUAFs0= 8, 25 -- Received: by 10.48.223.10 with SMTP id v10mr796858nfg; 8, 25 -- Thu, 09 Feb 2006 06:35:56 -0800 (PST) 8, 25 -- Received: by 10.49.55.19 with HTTP; Thu, 9 Feb 2006 06:35:56 -0800 (PST) 8, 25 -- Message-ID: {c4e021530602090635k847eadej4ffd50ec558f97c0-at-mail.gmail.com} 8, 25 -- Date: Thu, 9 Feb 2006 09:35:56 -0500 8, 25 -- From: John Cumings {cumings-at-umd.edu} 8, 25 -- Reply-To: cumings-at-umd.edu 8, 25 -- Sender: jcumings-at-gmail.com 8, 25 -- To: Microscopy-at-microscopy.com 8, 25 -- Subject: Wanted: Gatan 622SC 8, 25 -- MIME-Version: 1.0 8, 25 -- Content-Type: text/plain; charset=ISO-8859-1 8, 25 -- Content-Disposition: inline 8, 25 -- Content-Transfer-Encoding: 8bit 8, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k19EZv1W027008 ==============================End of - Headers==============================
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==============================
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==============================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.
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==============================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:
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==============================Original Headers============================== 15, 17 -- From bfoster-at-mme1.com Fri Feb 17 16:31:50 2006 15, 17 -- Received: from 5starpro.com (enterprise.5starpro.com [207.44.136.95]) 15, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1HMVnJe023336 15, 17 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:31:49 -0600 15, 17 -- Received: (qmail 19789 invoked from network); 17 Feb 2006 17:33:30 -0500 15, 17 -- Received: from host57-99.rancor.birch.net (HELO barbsd505.mme1.com) (65.17.57.99) 15, 17 -- by enterprise.5starpro.com with SMTP; 17 Feb 2006 17:33:30 -0500 15, 17 -- Message-Id: {7.0.1.0.0.20060217162833.01fc9ec0-at-mme1.com} 15, 17 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 15, 17 -- Date: Fri, 17 Feb 2006 16:31:54 -0600 15, 17 -- To: bagnell-at-med.unc.edu, Microscopy Listserver {microscopy-at-microscopy.com} 15, 17 -- From: Barbara Foster {bfoster-at-mme1.com} 15, 17 -- Subject: Re: [Microscopy] Floaters 15, 17 -- In-Reply-To: {200602172205.k1HM5BbH017110-at-ns.microscopy.com} 15, 17 -- References: {200602172205.k1HM5BbH017110-at-ns.microscopy.com} 15, 17 -- Mime-Version: 1.0 15, 17 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I had the same sludge problem about 1.5 years ago in a new Haskris chiller. Haskris recommends using only distilled water. That lasted about three months and the slime appeared. It was a combination of algae and small particles. Dumped the water and replaced with new distilled water and Skasol to flush. Then, new distilled water and one half liter of Hexid A4 from Applied Thermal Control Ltd. UK as supplied by SEM service tech. Chiller seized after about three months. Post mortem indicated that the impeller blades failed. Most likely due to misalignment of motor and pump.
Haskris replaced motor and pump assembly. Fluid was drained and replaced with distilled water and ethylene glycol (.5G to 4.5G distilled water). Filters were changed and no problems for about eight months. Liquid is not starting to become darker and small build up of stuff in chiller main filter. It is time to change liquid and filters. Main filter is in the water tank and is spec'd at about 50u. External toilet paper style filter is spec'd at 2u. So both get changed at the same time.
The Haskris unit specifically says to not use automotive antifreeze since it will deteriorate the BUNA N material in the chiller. Some anti freeze contains ethylene glycol. So I'm puzzled by the successful use of distilled water and EG. Perhaps they meant to say not to use 100% antifreeze rather than a diluted mix.
The other factor is that the SEM came with basically transparent water hoses. This is not good since the light gets into them and advances the algae. So this upcoming liquid and filter replacement will include replacing the hoses with opaque ones.
Overall, there are three aspects to be concerned about:
1. chiller guts and pump 2. hoses 3. SEM items that get chilled water (TMP, coils, etc.)
I don't think that there is a single simple answer to this problem since SEMs are different and chillers are different.
gary g.
At 02:04 PM 2/17/2006, you wrote:
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==============================Original Headers============================== 15, 20 -- From gary-at-gaugler.com Fri Feb 17 16:58:51 2006 15, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 15, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1HMwoD3002918 15, 20 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:58:51 -0600 15, 20 -- Received: (qmail 24885 invoked from network); 17 Feb 2006 14:58:50 -0800 15, 20 -- Received: by simscan 1.1.0 ppid: 24881, pid: 24883, t: 0.0890s 15, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 15, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 15, 20 -- by qsmtp3 with SMTP; 17 Feb 2006 14:58:50 -0800 15, 20 -- Message-Id: {6.2.3.4.2.20060217143542.0205ed40-at-mail.calweb.com} 15, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 15, 20 -- Date: Fri, 17 Feb 2006 14:58:51 -0800 15, 20 -- To: jfactor-at-ns.purchase.edu 15, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 15, 20 -- Subject: Re: [Microscopy] Re: Microscope cooling lines 15, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 15, 20 -- In-Reply-To: {200602172204.k1HM4d7s015164-at-ns.microscopy.com} 15, 20 -- References: {200602172204.k1HM4d7s015164-at-ns.microscopy.com} 15, 20 -- Mime-Version: 1.0 15, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
Just a couple of points I'd like to make w.r.t. your post.
I would not recommend using ethylene glycol as it is toxic. Propylene glycol is non-toxic.
The problem with using "antifreeze" mixtures not intended for use in these kinds of systems is that they do not usually contain the proper additives. Glycol solutions that are not HVAC grade will deteriorate over time through a type of polymerization that will plug things up and render the system inoperable. The resulting deposits thus formed are very inert and to my knowledge no one has ever found a way to clean them so you basically have to replace the chiller system.
HVAC grade polypropylene contains additives to avoid that problem plus inhibitors that stop corrosion of most commercially available materials in piping. That also includes seal materials, but I'm not sure about specifically N-buena seals. I'd have to look that up, but I would think it also compatible being such a commonly used seal material.
Biofouling is quite common in closed loop systems. Using a biocide is usually used in these systems to eliminate this problem.
The original poster to this thread likely is in a location where they have a professional water treatment company taking care of large HVAC systems. Perhaps they should talk with the representative of that company and find out what is being done for chemical treatment of chilled water systems there. They may be able to just get some of the proper chemicals that likely exist on-site already.
I would also like to point out, there is really no reason to go through the expense of using a glycol based system unless there is danger of freezing the coolant for some reason. There are numerous other water treatments that are much less expensive and work very well to keep a closed loop system running well. If there is little to no make up water needed for the closed system, once set-up properly, there is little more to do other than enjoy a clean running system...
dj
On Fri, 17 Feb 2006, gary-at-gaugler.com wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I had the same sludge problem about 1.5 years ago in } a new Haskris chiller. Haskris recommends using only } distilled water. That lasted about three months and } the slime appeared. It was a combination of algae and } small particles. Dumped the water and replaced with } new distilled water and Skasol to flush. Then, new } distilled water and one half liter of Hexid A4 from } Applied Thermal Control Ltd. UK as supplied by SEM } service tech. Chiller seized after about three months. } Post mortem indicated that the impeller blades failed. } Most likely due to misalignment of motor and pump. } } Haskris replaced motor and pump assembly. Fluid was } drained and replaced with distilled water and ethylene } glycol (.5G to 4.5G distilled water). Filters were } changed and no problems for about eight months. Liquid } is not starting to become darker and small build up of } stuff in chiller main filter. It is time to change liquid and } filters. Main filter is in the water tank and is spec'd } at about 50u. External toilet paper style filter is spec'd } at 2u. So both get changed at the same time. } } The Haskris unit specifically says to not use automotive } antifreeze since it will deteriorate the BUNA N material in } the chiller. Some anti freeze contains ethylene glycol. } So I'm puzzled by the successful use of distilled water and } EG. Perhaps they meant to say not to use 100% antifreeze } rather than a diluted mix. } } The other factor is that the SEM came with basically transparent } water hoses. This is not good since the light gets into them } and advances the algae. So this upcoming liquid and filter } replacement will include replacing the hoses with opaque ones. } } Overall, there are three aspects to be concerned about: } } 1. chiller guts and pump } 2. hoses } 3. SEM items that get chilled water (TMP, coils, etc.) } } I don't think that there is a single simple answer to this } problem since SEMs are different and chillers are different. } } gary g. } } } } At 02:04 PM 2/17/2006, you wrote: } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } My service engineer recommends neither tap nor distilled water, but } } rather bottled spring water. Has anyone yet mentioned the possibility } } that green sludge in the filter might be algae growing in the cooling } } water or the cooling unit? } } --Jan Factor } } } } --------------------------------------- } } Jan Robert Factor, Ph.D. } } Professor of Biology } } --------------------------------------- } } Natural Sciences } } Purchase College, State University of New York } } 735 Anderson Hill Rd. } } Purchase, NY 10577 } } USA } } --------------------------------------- } } Office Tel: 914-251-6659 } } Office Fax: 914-251-6635 } } E-mail: jfactor-at-ns.purchase.edu } } or- jan.factor-at-purchase.edu } } --------------------------------------- } } } } } } } } ==============================Original Headers============================== } } 4, 16 -- From jfactor-at-ns.purchase.edu Fri Feb 17 16:02:30 2006 } } 4, 16 -- Received: from zephyr.ns.purchase.edu } } (zephyr.ns.purchase.edu [199.79.168.193]) } } 4, 16 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id } } k1HM2PAx009529 } } 4, 16 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 } } 16:02:25 -0600 } } 4, 16 -- Received: from [10.52.0.79] ([10.52.0.79]) } } 4, 16 -- by zephyr.ns.purchase.edu (AIX5.1/8.11.6p2/8.11.0) } } with ESMTP id k1HM5lr17594; } } 4, 16 -- Fri, 17 Feb 2006 17:05:48 -0500 } } 4, 16 -- Message-ID: {43F647F0.1070409-at-ns.purchase.edu} } } 4, 16 -- Date: Fri, 17 Feb 2006 17:02:24 -0500 } } 4, 16 -- From: Jan Factor {jfactor-at-ns.purchase.edu} } } 4, 16 -- User-Agent: Thunderbird 1.5 (Windows/20051201) } } 4, 16 -- MIME-Version: 1.0 } } 4, 16 -- To: microscopy-at-microscopy.com } } 4, 16 -- Subject: Re: Microscope cooling lines } } 4, 16 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed } } 4, 16 -- Content-Transfer-Encoding: 7bit } } ==============================End of - Headers============================== } } } ==============================Original Headers============================== } 15, 20 -- From gary-at-gaugler.com Fri Feb 17 16:58:51 2006 } 15, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) } 15, 20 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1HMwoD3002918 } 15, 20 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 16:58:51 -0600 } 15, 20 -- Received: (qmail 24885 invoked from network); 17 Feb 2006 14:58:50 -0800 } 15, 20 -- Received: by simscan 1.1.0 ppid: 24881, pid: 24883, t: 0.0890s } 15, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 } 15, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) } 15, 20 -- by qsmtp3 with SMTP; 17 Feb 2006 14:58:50 -0800 } 15, 20 -- Message-Id: {6.2.3.4.2.20060217143542.0205ed40-at-mail.calweb.com} } 15, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 } 15, 20 -- Date: Fri, 17 Feb 2006 14:58:51 -0800 } 15, 20 -- To: jfactor-at-ns.purchase.edu } 15, 20 -- From: Gary Gaugler {gary-at-gaugler.com} } 15, 20 -- Subject: Re: [Microscopy] Re: Microscope cooling lines } 15, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} } 15, 20 -- In-Reply-To: {200602172204.k1HM4d7s015164-at-ns.microscopy.com} } 15, 20 -- References: {200602172204.k1HM4d7s015164-at-ns.microscopy.com} } 15, 20 -- Mime-Version: 1.0 } 15, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed } ==============================End of - Headers============================== }
==============================Original Headers============================== 12, 21 -- From "dljones-at-bestweb.net"-at-bestweb.net Fri Feb 17 20:43:13 2006 12, 21 -- Received: from mta4.srv.hcvlny.cv.net (mta4.srv.hcvlny.cv.net [167.206.4.199]) 12, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1I2hDlf008914 12, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 20:43:13 -0600 12, 21 -- Received: from localhost (ool-44c62883.dyn.optonline.net [68.198.40.131]) 12, 21 -- by mta4.srv.hcvlny.cv.net 12, 21 -- (Sun Java System Messaging Server 6.2-4.03 (built Sep 22 2005)) 12, 21 -- with ESMTP id {0IUV00D2N3JYUD00-at-mta4.srv.hcvlny.cv.net} for 12, 21 -- microscopy-at-microscopy.com; Fri, 17 Feb 2006 21:43:13 -0500 (EST) 12, 21 -- Date: Fri, 17 Feb 2006 21:43:12 -0500 (Eastern Standard Time) 12, 21 -- From: "David L. Jones" {"dljones-at-bestweb.net"-at-bestweb.net} 12, 21 -- Subject: Re: [Microscopy] Microscope cooling lines 12, 21 -- In-reply-to: {200602172305.k1HN5bhk012207-at-ns.microscopy.com} 12, 21 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 12, 21 -- To: gary-at-gaugler.com 12, 21 -- Cc: microscopy-at-microscopy.com 12, 21 -- Message-id: {Pine.WNT.4.64.0602172109370.408-at-dljtoshiba} 12, 21 -- MIME-version: 1.0 12, 21 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 12, 21 -- Content-transfer-encoding: 7BIT 12, 21 -- References: {200602172305.k1HN5bhk012207-at-ns.microscopy.com} ==============================End of - Headers==============================
If EG is toxic and PG is not, that is a factor. However, I do not care if one or the other is toxic. I do not swim in the 5 gallons of fluid. So, between the two glycols, which is best for SEM? I do not know. What do you think?
So there are toxic issues, corrosive issues, etc., etc. Well, then just dump the SEM, eh? No. I do not think so. One must work with the materials at hand. So, what do you think are the best materials for chiller fluid?
gary g.
At 06:43 PM 2/17/2006, you wrote: } Gary, } } Just a couple of points I'd like to make w.r.t. your post. } } I would not recommend using ethylene glycol as it is toxic. } Propylene glycol is non-toxic. } } The problem with using "antifreeze" mixtures not intended for use in } these kinds of systems is that they do not usually contain the } proper additives. Glycol solutions that are not HVAC grade will } deteriorate over time through a type of polymerization that will } plug things up and render the system inoperable. The resulting } deposits thus formed are very inert and to my knowledge no one has } ever found a way to clean them so you basically have to replace the } chiller system. } } HVAC grade polypropylene contains additives to avoid that problem } plus inhibitors that stop corrosion of most commercially available } materials in piping. That also includes seal materials, but I'm not } sure about specifically N-buena seals. I'd have to look that up, but } I would think it also compatible being such a commonly used seal material. } } Biofouling is quite common in closed loop systems. Using a biocide } is usually used in these systems to eliminate this problem. } } The original poster to this thread likely is in a location where } they have a professional water treatment company taking care of } large HVAC systems. Perhaps they should talk with the representative } of that company and find out what is being done for chemical } treatment of chilled water systems there. They may be able to just } get some of the proper chemicals that likely exist on-site already. } } I would also like to point out, there is really no reason to go } through the expense of using a glycol based system unless there is } danger of freezing the coolant for some reason. There are numerous } other water treatments that are much less expensive and work very } well to keep a closed loop system running well. If there is little } to no make up water needed for the closed system, once set-up } properly, there is little more to do other than enjoy a clean running system... } } dj } } On Fri, 17 Feb 2006, gary-at-gaugler.com wrote: } } } } } } } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
==============================Original Headers============================== 9, 21 -- From gary-at-gaugler.com Fri Feb 17 20:51:47 2006 9, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 9, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1I2pkQd018305 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 17 Feb 2006 20:51:47 -0600 9, 21 -- Received: (qmail 22693 invoked from network); 17 Feb 2006 18:51:46 -0800 9, 21 -- Received: by simscan 1.1.0 ppid: 22690, pid: 22691, t: 0.2511s 9, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 9, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 9, 21 -- by qsmtp3 with SMTP; 17 Feb 2006 18:51:46 -0800 9, 21 -- Message-Id: {6.2.3.4.2.20060217184632.02068418-at-mail.calweb.com} 9, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.3.4 9, 21 -- Date: Fri, 17 Feb 2006 18:51:46 -0800 9, 21 -- To: "David L. Jones" {"dljones-at-bestweb.net"-at-bestweb.net} 9, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 21 -- Subject: Re: [Microscopy] Microscope cooling lines 9, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 21 -- In-Reply-To: {Pine.WNT.4.64.0602172109370.408-at-dljtoshiba} 9, 21 -- References: {200602172305.k1HN5bhk012207-at-ns.microscopy.com} 9, 21 -- {Pine.WNT.4.64.0602172109370.408-at-dljtoshiba} 9, 21 -- Mime-Version: 1.0 9, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This is a perennial problem, I know, but does anyone have any effective "pet" solutions to the problem of charging and subsequent electrostatic arcing of TEM film in the extremely dry air we get in New England typically in the winter, and some other people get at other seasons of the year?
The discharges can occur at almost any stage of the handling, from getting the film out of the vendor's packing, loading and unloading in the microscope carriers and loading in the developing racks (we use Lucite ones). We have considered a humidifier in the darkroom, but that would involve blocking off the ventilation (we have make-up air positively blown into to room and exhaust actively sucked out) to prevent our moisture from being simply blown away.
Any hints would be welcome, but switching to digital imaging is one that is not going to happen.
Tony.
************************************* Anthony J. Garratt-Reed M.A. D.Phil. MIT Room #13-1027 77 Massachusetts Avenue Cambridge, Massachusetts 02139-4307 USA
Below are responses (abbreviated) to my question about which to use, copper or PVC, to replace water lines running from cooling units to microscopes. I will run any suggestions by my service engineers just to make sure there are no concerns regarding specific equipment. I do know that water pH is important and that may play a role in determining what additives to use:
I am a service engineer who works on electron microscopes for the last 17 years and my point of view: I would go with copper this way no matter what may or may not happen to the lines (cleaning out - whatever) you have no worries. (Ray Spengler)
I do not think that there will be a problem using copper or PVC. But you chould check with the eq maker to see if they have any issues. We use Copper and have not had any problems. We do get build up over time due to the hoses "rotting" over time and general scale buildup. (Karl Weiss)
USE PVC and make the final connection to the equipment with two coils of rubber hose for vibration control. ( Fran Laabs)
For our new IMAGE building, I specified PVC piping for the closed circ loop between the water chillers and scopes. We still notice a greenish sludge building up in the water filters (takes about 6-8 months to become significant) but I am certain that this is coming from the EM (copper cooling coils and iron connections---} electrolytic reaction). The EM service people told us that if we ever used acid to clean the lines that they would no longer warranty the microscope. The PVC lines are perfectly clean. (J. Bozzola)
Another material you may wish to consider is called PEX, which is a cross-linked polyethylene. I don't know too much about its characteristics, except that it's very smooth inside, which should retard crud accumulation and it's more opaque than white pvc. It may be worth checking out. (P. Grover)
As an FYI, if you have copper based and iron based materials mixed in the same system, the iron will corrode preferentially through galvanic coupling, not the copper. In fact, the iron becomes a sacrificial anode protecting the copper from corrosion. Any time you have those two materials in the same system, you should have dielectric couplings between the two or you will actively corrode the ferrous based material. Regarding acid cleaning, you should know what your deposits are in your piping before deciding how to clean them. As an FYI, copper will generally corrode at pH's below about 6.3 or so. There are low pH cleaners that can be used with copper, but they contain corrosion inhibitors. (D. Jones)
If you intend to clean the lines with acid, I suggest PVC, since Cu can be etched at low pH. In addition to floating some dichlorophene for algal control, we add a corrosion inhibitor. We have been using a Mo-based formula, which was available from Aqua Labs on the East coast and from Skasol on the West coast, so find a distributor in your area. I think that either Aqua or Skasol would be able to give you that info.(B. Tivol)
Along these lines, I noticed a lot more copper leaching into the lines when I used deionized water than regular or filtered tap water. Secondly, Why hasn't anyone suggested using antifreeze? Antifreeze intended for cars should have corrosion inhibitors and should be suitable for this purpose. I have still gotten green particles regardless of whether antifreeze was used or not and have regularly made it a practice to clean the filters in the lines once a semester. (Jerry Calvin)
I have used the ethylene/glycol/water mix through clear PVC piping between my circulator and TEM and vacuum coating unit for six years now. There is a long run of the piping in daylight. Absolutely no algae Problem. Use 50/50 ethylene glycol*/water and PVC pipes and you will never have a problem again. Make sure that the chiller/recirculator manufacturer OKs the use of ethylene glycol but I wouldn't anticipate any problem. Even 20/80 EG/Water will work well. (Ted Dunn)
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www3.agriculture.purdue.edu/microscopy
==============================Original Headers============================== 17, 21 -- From dsherman-at-purdue.edu Sat Feb 18 10:40:08 2006 17, 21 -- Received: from exchange.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 17, 21 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1IGe8dl014778 17, 21 -- for {microscopy-at-microscopy.com} ; Sat, 18 Feb 2006 10:40:08 -0600 17, 21 -- Received: from EXCH02.purdue.lcl ([128.210.63.234]) by exchange.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 17, 21 -- Sat, 18 Feb 2006 11:40:08 -0500 17, 21 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH02.purdue.lcl ([128.210.63.237]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 17, 21 -- Sat, 18 Feb 2006 16:40:08 +0000 17, 21 -- User-Agent: Microsoft-Entourage/11.2.1.051004 17, 21 -- Date: Sat, 18 Feb 2006 11:40:07 -0500 17, 21 -- Subject: Cooling lines-responses 17, 21 -- From: Debby Sherman {dsherman-at-purdue.edu} 17, 21 -- To: "message to: MSA list" {microscopy-at-microscopy.com} 17, 21 -- Message-ID: {C01CB817.D64C%dsherman-at-purdue.edu} 17, 21 -- Thread-Topic: Cooling lines-responses 17, 21 -- Thread-Index: AcY0qfmyOEGiJKCdEdqwBAARJN08Mg== 17, 21 -- Mime-version: 1.0 17, 21 -- Content-type: text/plain; 17, 21 -- charset="US-ASCII" 17, 21 -- Content-transfer-encoding: 7bit 17, 21 -- X-OriginalArrivalTime: 18 Feb 2006 16:40:08.0197 (UTC) FILETIME=[FA697350:01C634A9] ==============================End of - Headers==============================
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Email: soleimanij-at-tbzmed.ac.ir Name: Jafar Soleimani Rad
Organization: University
Title-Subject: [Filtered] scale bar
Question: Hi we are using A LEO 906 TEM, scale bar is not registered in the microfilms. We are having problem to calculating the real sizes. any help in this matter is appreciated.
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both rpowell-at-nanoprobes.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [Filtered] Re: [Microscopy]: NTA-nanogold for His tagged protein in EM?
Question: [Commercial disclaimer - I work for Nanoprobes, and we make NTA-Ni(II)-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
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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....
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, 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 **********************************************
==============================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==============================
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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
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Tue Feb 21 18:39:28 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 (8.12.11/8.12.8) with ESMTP id k1M0dSPK023528 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 21 Feb 2006 18:39:28 -0600 4, 22 -- Received: from localhost (water-dog [192.168.1.26]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id 63DDE359F6 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 21 Feb 2006 16:39:28 -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 AF21035C10 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 21 Feb 2006 16:39:16 -0800 (PST) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v623) 4, 22 -- In-Reply-To: {200602182232.k1IMW6Zs027921-at-ns.microscopy.com} 4, 22 -- References: {200602182232.k1IMW6Zs027921-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {7bf40a7674c34bd31ebb3e601deaaae1-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] viaWWW: scale bar 4, 22 -- Date: Tue, 21 Feb 2006 16:47:18 -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==============================
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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?
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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 **********************************************
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==============================Original Headers============================== 23, 27 -- From milesd-at-us.ibm.com Tue Feb 21 20:05:07 2006 23, 27 -- Received: from e3.ny.us.ibm.com (e3.ny.us.ibm.com [32.97.182.143]) 23, 27 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1M257Q0001451 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 20:05:07 -0600 23, 27 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 23, 27 -- by e3.ny.us.ibm.com (8.12.11/8.12.11) with ESMTP id k1M255Vc016162 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 21:05:05 -0500 23, 27 -- Received: from d01av01.pok.ibm.com (d01av01.pok.ibm.com [9.56.224.215]) 23, 27 -- by d01relay04.pok.ibm.com (8.12.10/NCO/VERS6.8) with ESMTP id k1M2562i220526 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 21:05:06 -0500 23, 27 -- Received: from d01av01.pok.ibm.com (loopback [127.0.0.1]) 23, 27 -- by d01av01.pok.ibm.com (8.12.11/8.13.3) with ESMTP id k1M255LU008116 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 21:05:05 -0500 23, 27 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com [9.56.229.50]) 23, 27 -- by d01av01.pok.ibm.com (8.12.11/8.12.11) with ESMTP id k1M2540P008094 23, 27 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 21:05:04 -0500 23, 27 -- In-Reply-To: {200602202111.k1KLBUjW018639-at-ns.microscopy.com} 23, 27 -- Subject: Re: [Microscopy] spam filters or ?? 23, 27 -- To: Microscopy-at-MSA.Microscopy.Com 23, 27 -- X-Mailer: Lotus Notes Release 7.0 HF85 November 04, 2005 23, 27 -- Message-ID: {OF22CCB88F.52878652-ON8525711D.000B000E-8525711D.000B7274-at-us.ibm.com} 23, 27 -- From: Darrell Miles {milesd-at-us.ibm.com} 23, 27 -- Date: Tue, 21 Feb 2006 21:05:03 -0500 23, 27 -- X-MIMETrack: Serialize by Router on D01ML076/01/M/IBM(Release 6.5.4FP2 HF4|December 20, 2005) at 23, 27 -- 02/21/2006 21:05:04 23, 27 -- MIME-Version: 1.0 23, 27 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
I'm looking for a reliable scanner for TEM Micrographs. For the past 4 years I used an old HP scanner that was very good with 35mm and TEM micrographs, unfortunate this scanner is no longer working, now I need a scanner that can capture a reliable image for routine everyday micrographs; in order to shorten darkroom time. Any subjection?
Thanks in advance
Omayra Velez Life Cell Branchburg, NJ
==============================Original Headers============================== 5, 18 -- From mayas003-at-yahoo.com Tue Feb 21 22:58:21 2006 5, 18 -- Received: from web84109.mail.dcn.yahoo.com (web84109.mail.dcn.yahoo.com [209.73.179.120]) 5, 18 -- by ns.microscopy.com (8.12.11/8.12.8) with SMTP id k1M4wK2N012775 5, 18 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 21 Feb 2006 22:58:20 -0600 5, 18 -- Received: (qmail 63646 invoked by uid 60001); 22 Feb 2006 04:58:20 -0000 5, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 5, 18 -- s=s1024; d=yahoo.com; 5, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 5, 18 -- b=l3I3qltvjW7C7MFLKskZiUVjJbPQ1cydee1f6xuI3x0YEOtyb1XLN+xZX7pO4ne8HOWyjClZfM4jfomwRvPZ1aIimFvPffyyv5mgzkS65fZVHYy/qLK30Kq3Ft6gNcfeVuPoHIlCg5oBNKSj3juJH9BgGJ/Hx0brg5OPd0fZ8gY= ; 5, 18 -- Message-ID: {20060222045820.63644.qmail-at-web84109.mail.dcn.yahoo.com} 5, 18 -- Received: from [68.239.240.252] by web84109.mail.dcn.yahoo.com via HTTP; Tue, 21 Feb 2006 20:58:20 PST 5, 18 -- Date: Tue, 21 Feb 2006 20:58:20 -0800 (PST) 5, 18 -- From: Omayra Velez {mayas003-at-yahoo.com} 5, 18 -- Subject: Scanner for TEM Micrographs 5, 18 -- To: Microscopy-at-MSA.Microscopy.Com 5, 18 -- MIME-Version: 1.0 5, 18 -- Content-Type: text/plain; charset=iso-8859-1 5, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Like microscopes, it really is the more you pay the better the result (even my kids £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
The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
I am not sure why these would not be legitimate. I have gotten such requests that cited my message of some date and some subject, so they already have my e-mail address. I can easily envision an anti-spam service where a person must respond to such a challenge to get added to a white-list of senders. I have replied to a couple of those posts as instructed and have not been asked to re-register.
However, I also sent them a note that such a service is not very compatible with membership in a list server like this. Everyone who ever posts to the list will get challenged to register their e-mail address. I doubt that many will. And if I declined to register and continued to get these registration requests, I would probably add that person to my junk mailers list.
If someone halfway around the world doesn't want to accept the knowledge that gets shared via this list then that is their problem. They should not use such a challenge system on the e-mail address they use for this or other listservers.
Thanks for extra interesting info. We actually have an elderly Agfa Duo-Scan T2550 here (your option 3) that cost £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
} Date: Tue, 21 Feb 2006 10:07:28 -0600 } To: heckman-at-bgnet.bgsu.edu } From: hyi-at-emory.edu } Reply-to: hyi-at-emory.edu } X-Resent-From: "Microscopy Listserver" {microscopy-at-microscopy.com} } Subject: [Microscopy] Re: TEM: Cell culture preparation } X-lewp: MicroscopyListSpam NAGS } X-MASF: 0.00% } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
-- __ Carol A. Heckman, Ph.D. Professor of Biological Sciences Director, Center for Microscopy & Microanalysis Bowling Green State University, Bowling Green, OH 43403 fax: (419) 372-2024 email: heckman-at-bgnet.bgsu.edu http://www.bgsu.edu/departments/biology/facilities/MnM ___________________________________________________________________________
==============================Original Headers============================== 6, 17 -- From heckman-at-bgnet.bgsu.edu Wed Feb 22 14:22:27 2006 6, 17 -- Received: from smtp01.bgsu.edu (smtp01.bgsu.edu [129.1.5.17]) 6, 17 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1MKMRnl027386 6, 17 -- for {microscopy-at-microscopy.com} ; Wed, 22 Feb 2006 14:22:27 -0600 6, 17 -- Received: from [129.1.85.81] (dhcp-85-81.bgsu.edu [129.1.85.81]) 6, 17 -- by smtp01.bgsu.edu (Switch-3.1.6/Switch-3.1.6) with ESMTP id k1MKMODY023311 6, 17 -- for {microscopy-at-microscopy.com} ; Wed, 22 Feb 2006 15:22:26 -0500 (EST) 6, 17 -- Mime-Version: 1.0 6, 17 -- X-Sender: heckman-at-mailstore.bgsu.edu 6, 17 -- Message-Id: {p04320404c022a24abe97-at-[129.1.85.81]} 6, 17 -- Date: Wed, 22 Feb 2006 15:23:42 -0800 6, 17 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 6, 17 -- From: Carol Heckman {heckman-at-bgnet.bgsu.edu} 6, 17 -- Subject: Fwd: [Microscopy] Re: TEM: Cell culture preparation 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 k1MKMRnl027386 ==============================End of - Headers==============================
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
==============================Original Headers============================== 6, 19 -- From DRK-at-SHCC.org Wed Feb 22 16:17:50 2006 6, 19 -- Received: from shcc.org (mail.shcc.org [64.213.211.200]) 6, 19 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1MMHnKX024752 6, 19 -- for {microscopy-at-microscopy.com} ; Wed, 22 Feb 2006 16:17:50 -0600 6, 19 -- Received: from DRK2 ("port 2599"-at-[64.213.211.3]) by SHCC.org (PMDF V6.1 #37411) 6, 19 -- with ESMTPA id {01LZ9SWBRN1M0005QC-at-SHCC.org} for microscopy-at-microscopy.com; 6, 19 -- Wed, 22 Feb 2006 14:20:49 -0800 (PST) 6, 19 -- Date: Wed, 22 Feb 2006 14:14:24 -0800 6, 19 -- From: Doug Keene {DRK-at-SHCC.org} 6, 19 -- Subject: fixation of intact e18.5 mouse limbs 6, 19 -- To: microscopy-at-microscopy.com 6, 19 -- Reply-to: DRK-at-SHCC.org 6, 19 -- Message-id: {01LZ9SWBUP8S0005QC-at-SHCC.org} 6, 19 -- MIME-version: 1.0 6, 19 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 6, 19 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 6, 19 -- Content-type: text/plain; charset=us-ascii 6, 19 -- Content-transfer-encoding: 7BIT 6, 19 -- Thread-Index: AcY3/VYRJ74GcEsESWuSwdf1DPWQqA== ==============================End of - Headers==============================
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
==============================Original Headers============================== 4, 25 -- From walck-at-southbaytech.com Wed Feb 22 17:43:41 2006 4, 25 -- Received: from ylpvm12.prodigy.net (ylpvm12-ext.prodigy.net [207.115.57.43]) 4, 25 -- by ns.microscopy.com (8.12.11/8.12.8) with ESMTP id k1MNhfVl022463 4, 25 -- for {Microscopy-at-microscopy.com} ; Wed, 22 Feb 2006 17:43:41 -0600 4, 25 -- Received: from pimout6-ext.prodigy.net (pimout6-int.prodigy.net [207.115.4.22]) 4, 25 -- by ylpvm12.prodigy.net (8.12.10 outbound/8.12.10) with ESMTP id k1MNhd3U006295 4, 25 -- for {Microscopy-at-microscopy.com} ; Wed, 22 Feb 2006 18:43:39 -0500 4, 25 -- X-ORBL: [64.169.193.90] 4, 25 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 4, 25 -- by pimout6-ext.prodigy.net (8.13.4 outbound domainkey aix/8.13.4) with ESMTP id k1MNhZko156844 4, 25 -- for {Microscopy-at-microscopy.com} ; Wed, 22 Feb 2006 18:43:40 -0500 4, 25 -- From: "Scott Walck" {walck-at-southbaytech.com} 4, 25 -- To: {Microscopy-at-microscopy.com} 4, 25 -- Subject: ROHS and European microscopists who are fishermen (or fisherwomen) 4, 25 -- Date: Wed, 22 Feb 2006 15:43:47 -0800 4, 25 -- Message-ID: {001701c63809$d6021680$7801a8c0-at-dynamicbl8uno3} 4, 25 -- MIME-Version: 1.0 4, 25 -- Content-Type: text/plain; 4, 25 -- charset="us-ascii" 4, 25 -- Content-Transfer-Encoding: 7bit 4, 25 -- X-Priority: 3 (Normal) 4, 25 -- X-MSMail-Priority: Normal 4, 25 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 4, 25 -- Importance: Normal 4, 25 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 ==============================End of - Headers==============================
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.
------------------------------------------------------------------------ ---- The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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.
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 a Bachelor or a Master's degree. The following competencies are required for both jobs: innovation in solving challenging problems; good communication and interpersonal skills; teamwork skills and results orientation.
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==============================
At 17:45 -0600 22/02/06, walck-at-southbaytech.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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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============================== 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:
1. Unsubscribe from the list, instead of annoying posters with 'out-of-office' automatic responses.
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
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============================== 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==============================
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] 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
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 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
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both rpowell-at-nanoprobes.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Organization: Nanoprobes, Incorporated
Title-Subject: [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==============================
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