I was wondering if anyone could tell me if addition of caffeine to the fixative affects antigenicity, generally speaking. I understand it's good for dealing with plant materials high in polyphenolic compounds. Does anyone know exactly what it's doing to the polyphenolics? Thanks, Karen
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dhaslam-at-kmslawyers.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, January 1, 2003 at 18:45:55 ---------------------------------------------------------------------------
Email: dhaslam-at-kmslawyers.com Name: donald haslam
Organization: kornfeld mackoff silber
Education: Undergraduate College
Location: vancouver bc canada
Question: I am an amateur botanist interested in studying rhododendron seeds for the purposes of identification of species and sub-species. I will be using a trinocular microscope of 10x to 40x magnification. If I use a digital camera, what is the recommended minimum number of pixels that I should insist on to usefully record images? Thank you. don p.s. How useful in such endevour is a zoom microscope?
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
] FROM BRo/johnson junior IVORY COST WEST AFRICA PHONE225 07464760 A CRY FOR HELP Dear Sir,
It is my pleasure to write you after much consideration since telephone communication can not be suitable enough to communicate to you at first. Being the only son of my father, late Chief johmson SMITH from KWAZULU NATAL in Republic of South Africa (SA) I am 18 years of age. My father was limited liability Cocoa and Gold merchant in JOHANNESBURG South Africa before his untimely death. After his business trip to Abidjan -C?te d'Ivoire, to negotiate on a cocoa and gold business he wanted to invest in Abidjan - C?te d'Ivoire. A Week after he came back from Abidjan, he was attacked with my mother by unknown assassins, which my mother died instantly but my father died after five days in a private hospital on that faithful afternoon. I didn't know that my father was going to leave me after I had lost my mother. But before he gave up the ghost, it was as if he knew he was going to die. He my father, MAY HIS SOUL REST IN PERFECT PEACE he disclosed to me that he deposited the sum of $15,800,000,00 US Dollars (FIFTEEN MILLION EIGHT HUNDRED THOUSAND DOLLARS) in a bank here in Abidjan- C?te d'ivoire.That the money was meant for his cocoa and Gold company he wanted to establish in Abidjan - C?te d'Ivoire though, according to my father he deposited the money in a confident account of the bank and he handed to me all the relievant documents of the deposited fund and instructed me to seek for a reliable and trust worthy business partner for my life time investment abroad. Now I have succeeded in locating the bank here in Abidjan - C?te d'Ivoire. Now I am soliciting for your assistance to help me to transfer this money out from Abidjan to your safe account abroad so that we will invest it in any meaningful lucrative business in your country because this is my only hope in life. Awaiting anxiously to hear from you so that we can discuss the modalities of this transaction. Please kindly contact me through email immediately for more discussion or call me on225-07464760. Thanks for your kind attention.
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA22730 for dist-Microscopy; Thu, 2 Jan 2003 10:24:49 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA22721 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Thu, 2 Jan 2003 10:24:18 -0600 (CST) Received: from novafrica.com ([193.251.130.55]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA22710 for {Microscopy-at-sparc5.Microscopy.Com} ; Thu, 2 Jan 2003 10:24:03 -0600 (CST) Received: (qmail 8412 invoked by uid 100); 2 Jan 2003 14:49:02 -0000
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FROM THE DESK OF;Engr CHRIST GESO, ABIDJAN COTE D,IVORIE WEST AFRICA PHONE 225 07464760 e-MAIL;christ.geso-at-ONDIKOI.COM
Dear sir,
On behalf of group of IVORY COST Senators i hereby propose the above to you.
I am Engr CHRIST.I.C.GESO Chairman Senate Committee on Contracts Review and foreign payments. There is presently available the sum of US$50m {Fifty Million United States Dollars} which members of this committee wish to transfer into your account, to be used for our re-election in 2003 general elections in IVORY COST
SOURCE OF FUND. This amount was realized from inflated or over-invoiced aspect of contracts executed by some foreign firms in 1999 when IVORUY COST hosted the world youth soccer Championship.ln the course of our duty we observed that there were some un-paid claims.lt is one of them that we intend to lodge into your bank account for our mutual benefit.
REMUNERATION. We have unanimously agreed that you will be entitled to 30% of the total sum, while 10% is set aside to offset expenses incurred during the transaction.
INVESTMENT. You will be required to place 60% of the total sum into any high yield investment facility in your country, for an initial duration of 8 months. During the 2003 elections, you will return 30% to us, but this time as a foreign loan for our election purposes. The accrued interest and the remaining 30% is what you will re-invest in the same process for a period of 4 years.
OPERATIONAL MODALITIES. We shall present you as one of the contractors awaiting payment for over-due contract payment for job executed for the Federal RepubliQU DE COTE,D,IVOREA in 1999.Application for claims, processing of approvals will be undertaken by my committee in conjunction with some highly placed officials, to ensure that fund is wired into your nominated bank account.
REQUIREMENTS. Your company details, banking details, as well as your confidential tel and fax numbers should be sent to me immediately to indicate your interest.
CONCLUSION. You do not stand any risk at all for being parts of this project, and your present line of business or profession is no hindrance. Honestly, our re-election in 2003 will be determined by this transaction and your ability to partner with us in all sincerity will enhance its success.
Note: That this transaction is expected to last between 7-14 days from the time we received this information, so i am waiting for your reply via my private e-mail address christ.geso-at-caramail.com MY PHONE NUMBER 225 07464760 Thanks for your co-operation and understanding while your urgent response is expected.
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id MAA23262 for dist-Microscopy; Thu, 2 Jan 2003 12:17:02 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id MAA23254 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Thu, 2 Jan 2003 12:16:31 -0600 (CST) Received: from fort-point-station.mit.edu (FORT-POINT-STATION.MIT.EDU [18.7.7.76]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id MAA23247 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 2 Jan 2003 12:16:19 -0600 (CST) Received: from grand-central-station.mit.edu (GRAND-CENTRAL-STATION.MIT.EDU [18.7.21.82]) by fort-point-station.mit.edu (8.9.2/8.9.2) with ESMTP id NAA06367; Thu, 2 Jan 2003 13:11:00 -0500 (EST) Received: from melbourne-city-street.mit.edu (MELBOURNE-CITY-STREET.MIT.EDU [18.7.21.86]) by grand-central-station.mit.edu (8.9.2/8.9.2) with ESMTP id NAA03292; Thu, 2 Jan 2003 13:10:46 -0500 (EST) Received: from emlab.mit.edu (EMLAB.MIT.EDU [18.82.0.121]) by melbourne-city-street.mit.edu (8.9.2/8.9.2) with ESMTP id NAA20435; Thu, 2 Jan 2003 13:10:46 -0500 (EST) Message-Id: {5.0.2.1.2.20030102111322.00ad1438-at-hesiod} X-Sender: tonygr-at-hesiod (Unverified) X-Mailer: QUALCOMM Windows Eudora Version 5.0.2
I have available two surplus X-ray analyzers (without detectors):
1: LINK/Oxford eX/L Mk. I, ca. 1988. Has been out of use for about 3 years, was in working order when last turned on. Has no video monitor. Has pulse processor, microscope scan control and electron image acquisition. Comes with SEM software suite.
2: LINK/Oxford Isis 300, ca. 1996. In use until recently. You provide the computer (works well with 300MHz Pentium, Win98 - it needs a computer with an ISA slot for the Translink card). Comes with Isis software suite on floppies, and key disks for TEM/STEM software suite (X-ray analysis, X-ray mapping, digital imaging, drift correction, etc). Has no work table.
Both have the original Link documentation.
These systems are offered on a strictly as-is basis, free to an academic user, you arrange the transport and take all responsibility. First-come, first-served. Easiest if you can visit Cambridge, MA and pick up. The Isis will fit into a family car. The eX/L might fit a car with a large trunk, but will certainly fit a station wagon or small pick-up.
I am not prepared to split these systems up for parts. If you want/need a specific board, then you take the whole thing and arrange disposal of the parts you do not need.
You can try calling with questions, but e-mail works well.
Tony Garratt-Reed.
** Anthony J. Garratt-Reed ** MIT Room # 13-1027 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
Does anyone have a good protocol for doing SEM on replicas (casts) of human skin on a living, breathing person. I presume one would use a silicon rubber. Can that be imaged directly or would one make an epoxy cast from it? This is our first foray into cosmetology.
Greg Gregory W. Erdos, Ph.D. Assistant Director, Biotechnology Program University of Florida P.O. Box 118525 Gainesville, FL 32611 352-392-1295
If you are looking to blot against naturally soluble proteins, then my first suggestion is to work with an assumption that during preparation of the tissue such proteins generally end up in the buffers, or the alcohols, etc. If not soluble, then the method that you must use to homogenize/subdivide any tissue to 'solubilize' portions of otherwise insoluble macromolecules should be sufficient to provide a useful specimen for blotting even if substantial crosslinking has occurred. By any standard, the blotting method that starts with fixed, processed and sectioned tissue (or vibratomed???) tissue should be compared at every step with fresh, unfixed and otherwise unprocessed tissue. IF the only difference between the section and the fresh tissue is HCHO, then that and its solvent constitute the only variables. You can compare fresh with HCHO-solvent, and solvent alone to see the effect of each. It would seem that blotting is an appropriate method for such determinations when assaying either Ab dilutions or epitope availability/presence.
Please remember the following. The anatomist looks at the bone and wonders where the molecule is. The biochemist looks at the molecules and wonders how they were assembled to form the bone. The histochemist looks at the molecule in the bone and wonders if that's where it was when the bone was still part of the living organism. For insoluble molecules the answers appear much more apparent (e.g., glycogen!).
Forgetting for the moment about "soluble or insoluble". If fixation was permitted to proceed for days beyond 24hr, then you are working with tissue in which there will be at least some frequency of methylene bridges linking adjacent molecules together. I am not very experienced with blotting in any case, but I have some familiarity with the methods. If the epitope for which you are going to assay has been tested on fixed tissue, with the same monoclonal you will use for blotting, without a requirement for retrieval, then the homogenized tissue sections (I assume that is what you will do with them.) should have the exposed epitope as well. If retrieval is necessary for immunohistochemistry, then there is no reason to expect that the homogenizing procedure alone will not also 'retrieve' a substantial amount of the epitope. Controlling fixed sections against unfixed tissue sections for normal consituent proteins is problematic but doable if you have a torsion balance with which to weigh the starting sectioned material.
Returning to the soluble and insoluble point again. If one compares identical amounts of fixed and fresh liver and calculates the amount of protein per cell, one should find that there is substantially more protein per cell in the fresh tissue and that the amount of protein per cell in fixed tissue falls with duration of fixation until no more diffusibile protein can escape the matrix or none remains.
Just for the sake of argument. Years ago a method by which one could ostensibly remove bound HCHO from a section was 1N NaOH. While this method might remove an -MeHO group, it probably isn't more likely to remove a so-called methylene bridge (a cross link) than any other hydrolytic method. In fact, if one turned to look, dreamily, out the window during such a procedure, the section might magically disappear.
} From my point of view, your question poses its own experimental answer. In modern immunohistochemistry each Ag/Ab pair places its own set of demands on the operator of the method.
Hope this helps,
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: Beveridge, Mark J. [mailto:bevermj-at-peds.ufl.edu] Sent: Monday, December 30, 2002 9:56 AM To: 'Microscopy-at-MSA.Microscopy.Com'
i would like to know if there is any way to unfix tissue fixed in paraformaldehyde for immunoblotting?
Can anyone tell me of the fate of RMC, Inc.? I haven't seen anything from them in several years. We have an MT6000-XL ultramicrotome in our lab that's in need of some service. Any suggestions are welcome.
Richard Geissler Director, EM Laboratory Department of Pathology & Lab Medicine 859-257-5068 geissle-at-mail.uky.edu
Hi Donald, No zoom is desirable for specific photmicrography. Indeed, zooming is OK for photographing for show, but is disconcerting for quantitative characterizing. Zoom if you must, but insure that you capture images at the same magnification for your quantified studies. As to a; camera, I would wager that the new Nikon Coolpix 4500? would do the trick nicely. You can find good and reasonable adapters widespread on the new (the Nikon adapter is the highest priced of all). The most important determinant for a CCD camera is the purpose for which you purchase it. You must know its limits - there are few with film because the enlarging capacity is so great. Such is not the case with CCD images. You can zoom in 1-3X and see pixels. Scan a 2x2" 35mm transparency at 24-bits and you have a file of size 285MB. VERY BIG PRINTOUT!!! or WORTHLESS RESOLUTION WHEN THERE IS NO PRINTER FOR BILLBOARDS AVAILABLE. My Olympus CCD camera is a 1.3M-Pixel and provides great 5x7 color prints. If I use a CCD camera on a microscope, I have to be close to the publishing mag when I capture the image. Not so with film - though there are limits in this as well.
Here's a link: http://www.leubner.ch/index.html
Here's another to an encyclopedic source for seeds: http://hort.cabweb.org/SeedSci/Pdfs/ssr08075.pdf The above is for a PDF file so if you don't have the reader, you should get it free from Adobe.
I am not a botanist, so might I suggest you find one at a nearby college. Having some help in such an endeavor would likely lead to the collection of data that are publishable. Even a lawyer or a legal assistant should appreciate the prospect of starting with some notion of what work would be most productive.
Also, when you discover the best species for pie, please let me know so I can purchase a pack of the seed.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: dhaslam-at-kmslawyers.com [mailto:dhaslam-at-kmslawyers.com] Sent: Thursday, January 02, 2003 12:54 AM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (dhaslam-at-kmslawyers.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, January 1, 2003 at 18:45:55 ---------------------------------------------------------------------------
Email: dhaslam-at-kmslawyers.com Name: donald haslam
Organization: kornfeld mackoff silber
Education: Undergraduate College
Location: vancouver bc canada
Question: I am an amateur botanist interested in studying rhododendron seeds for the purposes of identification of species and sub-species. I will be using a trinocular microscope of 10x to 40x magnification. If I use a digital camera, what is the recommended minimum number of pixels that I should insist on to usefully record images? Thank you. don p.s. How useful in such endevour is a zoom microscope?
I followed the discussions concerning pregnancy and sem operation with interest in the past. Unfortunately, I did not save the replies as it seemed unlikely to occur in our lab. We currently have a pregnant occassional operator. At her request, the decision has been made to transfer sem work to another individual for the duration of the pregnancy.
When I mentioned that the list server has covered this topic in the past, I was asked if I the information was still available. I welcome any new infomation and hope that some one can pass along archived info.
Gregory W. Erdos wrote: ============================================================= Does anyone have a good protocol for doing SEM on replicas (casts) of human skin on a living, breathing person. I presume one would use a silicon rubber. Can that be imaged directly or would one make an epoxy cast from it? This is our first foray into cosmetology. ============================================================= I would reference my own (old) publications:
"Characterizing Cosmetic Effects and Skin Morphology by Scanning Electron Microscopy", presented at SCC Annual Educational Symposium, May 1975, St. Louis; J. Soc. Cosmet. Chemists, 27, 509-531 (November 1976).
If one does not have easy access to this journal, a "sample" of this type of work can be seen on the SPI Supplies website at URL http://www.2spi.com/catalog/spec_prep/a.html
While we were not the very first to ever replicate, in vivo, human skin, we were the first to publish a testing method for evaluating skin care products for the cosmetics and toiletries industries, and later for the pharmaceutical industry.
For "cosmetology" which I assumed is for the substantiation of cosmetics industry advertising claims, one must use a very rapidly curing resin for the original cast or "negative" replica, otherwise the cast itself, and the occlusiveness of the covering creates changes ("moisturing effects" if you will) that are greater than the effects of the products being studied. Hence the need for a replicating system that cures in literally seconds, instead of minutes or even hours. The system we developed is what has evolved over the years into the SPI Supplies Wet Replica Kit, see URL http://www.2spi.com/catalog/spec_prep/wet_rep_kits.shtml
Can it be imaged directly? Well, yes it can but it is not the preferred way . It is far less confusing to make a positive replica from the negative, and photograph the positive. That is why the replicating kit described above comes with a pololefinic powder that so far as we are concerned makes the very best positives with the least amount of shrinkage or distortion.
Does it pass the test of being "pretty good?" I think so since many of the leading laboratories in the cosmetics and toiletries industry use this kit for their work. It has been pretty extensively peer-reviewed by a number of technical persons working on skin care products and "moisturizer" products.
I would be happy to elaborate more on this technique if there were any questions, either on or off the listserver.
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 ######################## ============================================
Greg Dental silicone is excellent for replicating skin. The product I am familiar with is Kerr Reflect, though there are others no doubt. The setting time at body temperature is rapid, the material is comparatively non-toxic, releases very cleanly and the replica contains information with resolution approaching 100nm. Positive replicas can be made with epoxy, but I have had better results using hot-melt plastics such as polycarbonate. My method was fairly unrefined - the silicone negative replica was placed in an oven at just above the melting point of the plastic, a piece of 3mm plastic sheet placed on top and weighted with a brass weight. PTFE or silicone sheet was used to prevent adhesion between plastic and weight. The specimens were allowed to cool to room temperature with the weights in place before separating positive from negative. With care more than one copy can be made if necessary, though I daresay fine detail is gradually degraded by this. I think the main reason this has worked better for me than replicating with epoxy is that I use it for replicating leaves, which are covered with wax crystals. The silicone picks up these waxes, so epoxy only sees their undersides, especially when cold- polymerized. At high temperature, polycarbonate displaces the molten wax and can replicate the negative replica of the outer surface. This may not be an issue for you in replicating skin, but the combination of high temperature and pressure also helps displace air from features like hairs, which epoxy can be reluctant to enter. Vacuum infiltration of epoxy at high temperature may improve this though.
Hope this is of some use. Best wishes for the new year Chris
On 2 Jan 03, at 15:39, Greg Erdos wrote:
} ---------------------------------------------------------------------- } -- The Microscopy ListServer -- Sponsor: The Microscopy Society of } America To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Fellow scopesters } } Does anyone have a good protocol for doing SEM on replicas (casts) of } human skin on a living, breathing person. I presume one would use a } silicon rubber. Can that be imaged directly or would one make an } epoxy cast from it? This is our first foray into cosmetology. } } Greg } Gregory W. Erdos, Ph.D. } Assistant Director, Biotechnology Program } University of Florida } P.O. Box 118525 } Gainesville, FL 32611 } 352-392-1295 } }
========================================== Dr. Chris Jeffree University of Edinburgh BIOSEM - Biological Sciences Electron Microscope Facility Institute of Cell and Molecular Biology Waddington Building King's Buildings, Mayfield Road EDINBURGH, EH9 3JN, Scotland, UK Tel. #44 (0) 131 650 5554 /8669 FAX. #44 (0) 131 650 6563 Mobile 07710 585 401 email c.jeffree-at-ed.ac.uk =========================================
Here is the table of contents for the January '03 issue of Microscopy Today:
Carmichael: Out with the Old, In with the New Boyes: Pros and Cons of Low Voltage SEM EDX Elemental Analysis Basgen: Basic Stereology Sedgewick: Segmentation Before Quantization By Using Photoshop: Darkfield Images Clarke: Rediscovery of Darkfield Dispersion Staining while Building a Universal Student Microscope Beanland: Rapid Cross-Section TEM Specimen Prep. of III-V Materials Harmsen: Scan Speed, Mag and Accuracy. That is the Question! Maleeff: Removing Very Fine Wrinkles from Half-Micron Epoxy Sections Killius: Problems with Hydrophobicity of Coated Grids Mascorro: The Versatile Family of Epoxy Resins: Designing Embedding Media with Specific Viscosity Properties Fandrich, et al.: Secondary Melt Ornamentation On Westwater, Utah: Microspherules: Evidence Of An Extraterrestrial Provenance
Some news about Microscopy Today: As of 12/31/02 we have 17,411 subscribers, making us the largest microscopy magazine in the USA.
I will close the subscription list for mailing the January issue on January 7th. You must subscribe before then to receive this issue.
All subscriptions/changes/unsubscribes must come via our website: www.microscopy-today.com
Note that if you live in the USA and received the November issue of MT, along with the M&M-2003 Call for Papers, you probably already have a subscription.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
I wish this my proposal will not come to you as a surprise. I am MIKE CACUS, a Director with an International Bank in San Pedro - Cote dŽIvoire formerly called Ivory Coast.
We had a foreign client (name with held) who deposited a huge sum of money (US$9.6 million United States dollars) with our company. Eventually, this client was among the victims of EGYPT AIR BOEING 767 FLIGHT N° 990 that crashed on the 31-10-1999 in USA, since then we have not had anybody coming for the claims as the next of kin. A situation I have monitored closely with my position in the company.
Now, haven monitored this deposit and managed it over the years before his death, and hence nobody has showed up as the next of kin for the past two year plus, I have removed the file to my private vault.
I now solicit for your assistance to present you as the next of kin as every other arrangement has been concluded by me and I am only waiting for a foreigner to enable me move the fund to his account. This does not have any risk attached to it as all the internal documentation will be handled by me.
I therefore request you to confirm your interest by a return message and I will furnish you with details. Your interest will be negotiable before we commence the operation.
You could use a dental impression material; they are usually silicone-based but can be other things, and all are completely harmless to the subject and the experimenter. It is possible to image the negative made this way, but there are problems with its behaviour under vacuum. The negative can be used to make a positive in epoxy resin which is virtually indistinguishable under SEM from the original.
Wieland M, Chehroudi B, Textor M, Brunette DM. Use of Ti-coated replicas to investigate the effects on fibroblast shape of surfaces with varying roughness and constant chemical composition. J Biomed Mater Res. 2002 Jun 5;60(3):434-44. PMID: 11920667 [PubMed - indexed for MEDLINE]
2: Chehroudi B, Brunette DM. Subcutaneous microfabricated surfaces inhibit epithelial recession and promote long-term survival of percutaneous implants. Biomaterials. 2002 Jan;23(1):229-37. PMID: 11762842 [PubMed - indexed for MEDLINE]
Hope this helps.
Lesley Weston.
on 02/01/2003 12:39 PM, Greg Erdos at gwe-at-biotech.ufl.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Fellow scopesters } } Does anyone have a good protocol for doing SEM on replicas (casts) of human } skin on a living, breathing person. I presume one would use a silicon } rubber. Can that be imaged directly or would one make an epoxy cast from } it? This is our first foray into cosmetology. } } Greg } Gregory W. Erdos, Ph.D. } Assistant Director, Biotechnology Program } University of Florida } P.O. Box 118525 } Gainesville, FL 32611 } 352-392-1295 } }
As I was the instigator of this subject, I thought that I would give you a synopsis of what I learned.
1. It is against Federal Law to ask any employee if they are pregnant, plan to get pregnant, or are sexually active (both male and female). 2. The lab is legally responsible for any damage which may occur to the mother or fetus due to working in the lab. So, the lab is responsible, and therefore the management liable, but it is against the law to know what condition your workers are in vis a vis reproduction. 3. What I have gone to here is treating everyone as if they are reproductive. With the appropriate protocols this is possible and makes for a much safer work environment for everyone. 4. The use of modern microscopes is not dangerous from radiation. If there is radiation coming out of your scope , get it fixed or get rid of it. There are many good scopes out there for a pittance. If you are really paranoid, get one of those badges and have your heaviest user wear it, if it show radiation, see above. 5. With the use of appropriate hoods and safety equipment, anyone should be able to prepare samples. Do however, use the safer chemical and/or procedure when possible. Such as Hepes, not Cacodylate; microwave fixation in hood, not conventional. 6. Once you know that you have a pregnant worker, contact your safety people and have the worker prepare a personal hygiene plan and have it approved by the safety office and suggest that her doctor have a look at it to see if he/she has any concerns. I had two pregnant workers in the lab at the same time, there were no problems.
Just one personal aside, it is much easier to manage an employee if you know that she is pregnant, some have little to no problems, but others can have a very hard time. If you do not know what is going on, the possibility is there to misjudge the performance of that individual, but the law allows for the worker to privacy. Just one more thing to deal with in the modern world.
Bill
William McManus Supervisor Microscopy Facility Utah State University Logan UT 84322-5305
-----Original Message----- } From: Cochran [mailto:fisher-at-meganet.net] Sent: Thursday, January 02, 2003 6:45 PM To: microscopy-at-sparc5.microscopy.com
Hi All,
I followed the discussions concerning pregnancy and sem operation with interest in the past. Unfortunately, I did not save the replies as it seemed unlikely to occur in our lab. We currently have a pregnant occassional operator. At her request, the decision has been made to transfer sem work to another individual for the duration of the pregnancy.
When I mentioned that the list server has covered this topic in the past, I was asked if I the information was still available. I welcome any new infomation and hope that some one can pass along archived info.
} 6. Once you know that you have a pregnant worker, contact your safety } people and have the worker prepare a personal hygiene plan and have it } approved by the safety office and suggest that her doctor have a look at } it to see if he/she has any concerns. } I had two pregnant workers in the lab at the same time, there were no } problems. } :
How long is it going to take to go through each step of the procedure mentioned above? By the end of that procedure it might be too late for the fetus.
I believe that as soon as a lady learned or suspect she is pregnant she should stop working with radiation stuff. There is no safe dose of radiation. Risk might be small but it is always positive.
Kind regards, Ayten.
} Just one personal aside, it is much easier to manage an employee if you } know that she is pregnant, some have little to no problems, but others } can have a very hard time. If you do not know what is going on, the } possibility is there to misjudge the performance of that individual, but } the law allows for the worker to privacy. Just one more thing to deal } with in the modern world. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } -----Original Message----- } } From: Cochran [mailto:fisher-at-meganet.net] } Sent: Thursday, January 02, 2003 6:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Pregnancy & SEM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } I followed the discussions concerning pregnancy and sem operation with } interest in the past. Unfortunately, I did not save the replies as it } seemed unlikely to occur in our lab. We currently have a pregnant } occassional operator. At her request, the decision has been made to } transfer sem work to another individual for the duration of the } pregnancy. } } When I mentioned that the list server has covered this topic in the } past, I was asked if I the information was still available. I welcome } any new infomation and hope that some one can pass along archived info. } } Thanx, } Ray Cochran } } } }
I'm fairly interested in this, too, would you please copy responses to me?
I don't know how others feel, but I'm always a bit disappointed when people ask for off-list replies, as it shuts me out of the loop, having given a tantalising initial glimpse of a thread. It seems to me that one of the wonderful things about this list is the discussions, and I would hate for it to be merely a bulletin board on which people posted their requests for information.
Commercial and personal privacy issues occasionally preclude open discussion, I understand that, but that's pretty rare.
Happy New Year
rtch
} } Hi, } } We have a researcher on campus who would like to examine the cell size } of 3000 year old wood. Any ideas on the sample preparation protocol to } preserve the cell structure without shrinking? } } Please direct your responss to Cheryl Jensen (jensenc-at-missouri.edu). } } Thanks in advance. } Lou Ross } -- } Senior Electron Microscope Specialist } Electron Microscopy Core Facility } W136 Veterinary Medicine } University of Missouri } Columbia, MO 65211-5120 } (573) 882-4777, fax 884=5414 } email: rosslm-at-missouri.edu } web: www.biotech.missouri.edu/emc }
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
Your pont is well taken, but the law does not let us take this kind of action. In the past, companies have used pragnancy as a tool to demean women, and even if meant in the best of intentions, a lab manager cannot get involved until it is probably too late, a catch 22. That is why, I have set up my lab so that all workers are treated the same, is if they are pregnant, even the men. An issue generally not discussed is, what are the ramifications to male gametes when exposed to chemical and radiation exposure? But again, let me repeat, if you have an EM that is leaking radiation that is bad, and should be dealt with, period. Fix it or pitch it. In reality, we are exposed to more teratigens and carcinogens in the grocery store than in a well run EM lab.
Bill
-----Original Message----- } From: celik ayten [mailto:celik-at-ews.uiuc.edu] Sent: Friday, January 03, 2003 1:20 PM To: William McManus Cc: fisher-at-meganet.net; microscopy-at-sparc5.microscopy.com
I was once told that the only possible way to get irradiated by an SEM was to have your body under the beam while it is operating. I have not tried it yet to see if it is true or not.
Ron L
-----Original Message----- } From: celik ayten [mailto:celik-at-ews.uiuc.edu] Sent: Friday, January 03, 2003 3:20 PM To: William McManus Cc: fisher-at-meganet.net; microscopy-at-sparc5.microscopy.com
On Fri, 3 Jan 2003, William McManus wrote:
[skip]
} 6. Once you know that you have a pregnant worker, contact your safety } people and have the worker prepare a personal hygiene plan and have it } approved by the safety office and suggest that her doctor have a look at } it to see if he/she has any concerns. } I had two pregnant workers in the lab at the same time, there were no } problems. } :
How long is it going to take to go through each step of the procedure mentioned above? By the end of that procedure it might be too late for the fetus.
I believe that as soon as a lady learned or suspect she is pregnant she should stop working with radiation stuff. There is no safe dose of radiation. Risk might be small but it is always positive.
Kind regards, Ayten.
} Just one personal aside, it is much easier to manage an employee if you } know that she is pregnant, some have little to no problems, but others } can have a very hard time. If you do not know what is going on, the } possibility is there to misjudge the performance of that individual, but } the law allows for the worker to privacy. Just one more thing to deal } with in the modern world. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } -----Original Message----- } } From: Cochran [mailto:fisher-at-meganet.net] } Sent: Thursday, January 02, 2003 6:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Pregnancy & SEM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } I followed the discussions concerning pregnancy and sem operation with } interest in the past. Unfortunately, I did not save the replies as it } seemed unlikely to occur in our lab. We currently have a pregnant } occassional operator. At her request, the decision has been made to } transfer sem work to another individual for the duration of the } pregnancy. } } When I mentioned that the list server has covered this topic in the } past, I was asked if I the information was still available. I welcome } any new infomation and hope that some one can pass along archived info. } } Thanx, } Ray Cochran } } } }
Chris Jeffree wrote: ========================================================== Dental silicone is excellent for replicating skin. The product I am familiar with is Kerr Reflect, though there are others no doubt. The setting time at body temperature is rapid, the material is comparatively non-toxic, releases very cleanly and the replica contains information with resolution approaching 100nm. Positive replicas can be made with epoxy, but I have had better results using hot-melt plastics such as polycarbonate. My method was fairly unrefined - the silicone negative replica was placed in an oven at just above the melting point of the plastic, a piece of 3mm plastic sheet placed on top and weighted with a brass weight. PTFE or silicone sheet was used to prevent adhesion between plastic and weight. The specimens were allowed to cool to room temperature with the weights in place before separating positive from negative. With care more than one copy can be made if necessary, though I daresay fine detail is gradually degraded by this. I think the main reason this has worked better for me than replicating with epoxy is that I use it for replicating leaves, which are covered with wax crystals. The silicone picks up these waxes, so epoxy only sees their undersides, especially when cold- polymerized. At high temperature, polycarbonate displaces the molten wax and can replicate the negative replica of the outer surface. This may not be an issue for you in replicating skin, but the combination of high temperature and pressure also helps displace air from features like hairs, which epoxy can be reluctant to enter. Vacuum infiltration of epoxy at high temperature may improve this though. ================================================================ The polymerization time of the Kerr "Reflect" system is longer than the system I described a few days ago as the SPI Wet Replica Kit. I agree that the faster polymerization time for the replication of leaves might not be important, and a longer time might even be at times an advantage. But once the silicone is applied to skin, it literally stops all transepidermal water loss. And the trapped water has no where to go but to build up in the stratum corneum (the outer most layer of skin, or the dead skin layer), creating an experimentally induced artifact effect. This effect is often times more profound than the subtle difference one is trying to discern in terms of product efficacy or whether one product is better than another. That is one reason why so many times projects of this type, where one is looking for differences fail because the transepidermal moisture buildup makes everything look the same!
We have not looked lately at this particular Kerr product but we have looked at other dental silicones and in order to impart to them the kind of dimensional stability needed for dental impressions, they are loaded with an inorganic additive system to give them that desired dimensional stability. When one starts going up in magnification, about about 500X, the additives from the negative replicating material can be resolved (as an artifact effect). The SPI Wet Replica Kits does not have to have such good dimensional stability, and therefore has a different type and loading of the inorganic additives, and one can generally achieve magnifications (before artifacts set in) about 1.5X higher, to about 750X. While this might not sound very high in magnification in SEM terms, it tends to be more than high enough for visualizing the effects of cosmetic and topically applied pharmaceutical products to human skin. It has been our experience that most meaningful work is done not higher than about 300X although for some kinds of work up to about 600X.
The advantage of a much faster polymerizing system is that the replica essentially "cures" before the body generated moisture buildup starts to create effects all of its own. And that is what makes possible the visualization of very subtle effects from the application of a product, either short term of long term.
The negatives made this way are understandably fragile but the non-wetting characteristics of the silicone on skin, even on dry flaky skin, leads to an easy lift-off without much in the way of disruption. We have found that on most types of skin, and in particular, dry skin, a first positive is made and thrown out; its function is to serve as a "cleaning replica", to clean off any remains of skin flakes. The second replica is what is then used for the microscopy. The low melting polyolefinic very fine powder used for the positive replicas will flow pretty easily into the cylindrical "holes" that would be representing a shaft of hair. The entrapment of air bubbles is not a serious problem.
We have found that for skin work, epoxies and just about anything else much more brittle than the polyolefin used for the positive does not work as well ; that way even curved channels and irregularly shaped volumes can be successfully transmitted to the positive replica where as if done in other materials, for example, an epoxy, such micro-volumes would be snapped off then the two replicas were separated.
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 ######################## ============================================
Dick I understand that the Kerr Reflect product range has been replaced by a new range known as Extrude. The closest equivalent to the Kerr Reflect Wash (low viscosity formulation) that I used is Kerr Extrude Wash. Check out the other impression materials in their range. There is, for example, a product based on sodium alginate that is aqueous, hydrophilic, and has faster setting times than the Kerr silicone. For details and suppliers contact Kerr Dental:
www.KERRDENTAL.com Mailing Address : 1717 West Collins Orange, CA 92867 Phone : (800) KERR-123 (800-537-7123), (714) 516-7400
Silicone impression materials are supplied by most dental laboratory supply dealers throughout UK, and probably in US too.
Chris
----- Original Message ----- } From: "Richard Gordon" {gordonr-at-Ms.UManitoba.CA} To: "Chris Jeffree" {cjeffree-at-srv0.bio.ed.ac.uk} Cc: "Richard Gordon" {gordonr-at-Ms.UManitoba.CA} Sent: Saturday, January 04, 2003 11:51 PM
Here is my reply to this, with some additions:
One way to do this would be to freeze the tissue in the cryo-prep chamber of cryoSEM, then observe frozen at low kV. Or plunge freeze in LN2, cryoplane, transfer to cryostage, etch, observe. If the wood is dry, freezing is probably unnecessary. And if you just want to look at cell size on a surface, you may get away without coating at very low kV. Depends on how much you are able to process this wood. Roger Heady - Roger.Heady-at-anu.edu.au, http://sres.anu.edu.au/people/headyr.html - is an expert in examining wood in SEM and could give much better advice - I don't think he's on this email list. cheers, Rosemary } } } Hi, Lou } } I'm fairly interested in this, too, would you please copy } responses to me? } } I don't know how others feel, but I'm always a bit disappointed } when people ask for off-list replies, as it shuts me out of the } loop, having given a tantalising initial glimpse of a thread. It } seems to me that one of the wonderful things about this list is } the discussions, and I would hate for it to be merely a bulletin } board on which people posted their requests for information. } } Commercial and personal privacy issues occasionally preclude } open discussion, I understand that, but that's pretty rare. } } Happy New Year } } rtch } } } } } } Hi, } } } } We have a researcher on campus who would like to examine the cell size } } of 3000 year old wood. Any ideas on the sample preparation protocol to } } preserve the cell structure without shrinking? } } } } Please direct your responss to Cheryl Jensen (jensenc-at-missouri.edu). } } } } Thanks in advance. } } Lou Ross } } -- } } Senior Electron Microscope Specialist } } Electron Microscopy Core Facility } } W136 Veterinary Medicine } } University of Missouri } } Columbia, MO 65211-5120 } } (573) 882-4777, fax 884=5414 } } email: rosslm-at-missouri.edu } } web: www.biotech.missouri.edu/emc } } } } Ritchie Sims Phone : 64 9 3737599 ext 7713 } Department of Geology Fax : 64 9 3737435 } The University of Auckland email : r.sims-at-auckland.ac.nz } Private Bag 92019 } Auckland } New Zealand
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Allen Sampson {ars-at-sem.com} To: "'Sergey Ryazantsev'" {sryazant-at-ucla.edu} , "microscopy-at-sparc5.microscopy.com" {microscopy-at-sparc5.microscopy.com} 12/22/02 05:26 AM cc: Please respond to Subject: RE: Evaporator cold trap "ars-at-sem.com"
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Sergey,
That is a strong pump for this system - the normally configured system uses
a 250 L/sec pump. A diffusion pump, as any pump, is a differential system - The ultimate pressure achieved is a balance between the leak rate in the input, the back pressure in the output and the bypass rate in the pump (how
much gas can pass from the output to the input for a given pressure differential, giving rise to the mechanical pump oil contamination in a system). Although these pumps are rated up to 10-9 pressures, this is never seen in operational systems. If you were to cap the diffusion pump with a metal sealed, solid metal cap and had an enormous backing pump, you may come close to the rated ultimate rating. In practical systems, these levels are but a dream.
I'd be interested in knowing what roughing pump you are using.
No vacuum system has no leaks, particularly when elastomeric seals are used. The standard Denton, with its manual valves and bell jar seal, has a
lot of elastomeric seals. And a cold cathode vacuum gauge accuracy is debatable at 10-6 or less.
I've only recommended and used Santovac 5 for over twenty years. It is a great diffusion pump oil. My main reason for recommending it is not the ultimate vacuum attainable (debatable considering the greater temperature apparently needed by Santovac, although I've never had a problem using it in any diffusion pump), but rather it's relative insensitivity to sudden air inrushes when hot. That means that it will 'crack' and polymerize less
than other oils. In other words - it will last longer and cause less hard deposits on the pump.
As far as o-rings, I've found Buna to be quite acceptable, lightly coated with Brayco, for static seals. Where dynamic seals are used (rotational or
translational forces are common), I use Buna with Apiezon (the waxier Apiezon has more staying power, although it will also hold particulate contaminants more). The reason for cracked o-rings is neglect, not the suitability of vacuum greases. The reason for greasing o-rings is to provide a light coating that preserves the qualities of the elastomeric material, not to make up for insufficiencies in the vacuum sealing surfaces. In this respect, the better vacuum greases do a good job and do not compromise either Buna or Viton. I have many 25+ year old systems that
have original o-rings that are indistinguishable from new, in appearance, shape or conformance.
The deformability of Buna is actually a plus, at least in systems that are mostly held at vacuum. You may have noticed that a system that was just rebuilt with new or rebuilt o-rings takes some time to come to an ultimate vacuum equilibrium. That, of course, involves the outgassing of the system
components after being exposed to atmosphere for some time during the rebuild. But it also includes the time required for the elastomeric vacuum
seals to be 'sucked' into place. A well designed o-ring seal will depend on the mechanical pressures on the o-ring, but will ultimately depend on the o-ring's conformance under the gas pressures it's subjected to.
In my experience, the Buna o-rings will deform to provide a good seal faster and, properly maintained, will continue to conform to a shape that best seals. I generally use Viton for it's improved resistance to high temperatures. In either case, I use an acetone wipe for cleaning o-rings every time I recondition them. It tends to swell the o-rings with two effects - it restores them to their original shape and helps to provide a quick seal when the system is pumped down again.
BTW, I've cleaned more DPs and refurbished more vacuum systems than I'd like to elucidate. In my business, I tend to get the instruments that the manufacturers don't service anymore, didn't service properly or have been neglected for some time.
Just a few ruminations from a long career of servicing many vacuum instruments.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
On Sunday, December 22, 2002 3:12 AM, Sergey Ryazantsev [SMTP:sryazant-at-ucla.edu] wrote: } Allen } } DV502A has 800 l/sec DP!!! It enough to pump down a huge volume. If you
} don't have any leaks in your system, the vacuum would be directly dependent } from the ultimate vacuum for the pump (which is somehow dependent from } pump's actual construction/quality and DP Oil). In most cases leaks are the } reason of vacuum degradation. Any normally serviced vacuum evaporator } should deliver at least very good 10-6 torr. 10-5 is very bad and actually } you may not use it for biological sample preparation. In my particular } case, I've replaced ALL O-rings on viton with touch of Apiezon L, cleaned
} and polished DP and used Santavac-5. Santavac-5 is great DP oil. You have } to try. On my DV502A I have 2*10-6 after about 40 min pumping (no LN2) and } better than 5*10-7 overnight. I am using MKS cold cathode gauge with } sensitivity up to 10-9 torr (which I don't need). I think, the vacuum } quality is mostly a function of how clean your system and how many 10-year } old cracked "buna" O-rings is inside your system. The problem with "buna" } - it does not hold the shape and easily deformed even if it's not old. It's } also destroyed by vacuum grease (does not matter what manufacturers told } you). Viton is much better. Since I spent $500 on Santavac-5 8 years ago, } I never touch my DP again. By the way, I have another vacuum system, which } I build by myself. It has exact the same volume and similar amount of } O-rings but 400 l/s TP from SEIKO. That system is oil-free. I mean, it } has scroll pump as a backing device and TP itself does not have any oil } (it's magnetically levitated beauty). So, when I build the system, I was
} expecting similar productivity for this system as for DV502A but } oil-free. I was completely wrong! This "baby" easily delivered to me } 5*10-7 torr in 20 (yes, twenty) min! After a few hours, it's going into } 10-8. So, my "theory" is that in the standard setup, oil from mechanical
} pump contaminated the whole system (and your sample!) and adsorbs a lot of } air, which slowly released during the high vacuum pumping cycle. Because
} my new system is oil-free, it's much faster. It has 12x12" Bell Jar with
} 6' collar. Another example: my old Polaron with 100 l/s DP (Santavac-5 } again) and 12x12" Bell Jar. This system is very comfortable with 2*10-6 } (no LN2) after I repaired manufacturers defect in DP. All tricks how to get } good vacuum perfectly described in the Wil Bigelow book. Have a great } Holidays (don't start cleaning DP- it's messy)! Sergey } } } At 08:13 PM 12/20/02 -0600, you wrote: } } Sergey, } } } } Have you verified that vacuum level with an independent, calibrated vacuum } } gauge? Vacuum sounds way to high for that system. I agree with all of } } your recommendations, but case in point, the vacuum you claim is on an } } order of what we normally find in a valve isolated electron gun (much } } smaller vacuum and seal volumes) pumped by a 20 L/sec ion pump. The vacuum } } level you stated for the start of your work is far closer to the best I } } have seen out of this particular evaporator after a complete rebuild. } } } } Allen R. Sampson } } Advanced Research Systems } } 317 North 4th. Street } } St. Charles, Illinois 60174 } } } } phone (630) 513-7093 fax (630) 513-7092 http://www.sem.com } } } } } } On Friday, December 20, 2002 3:01 PM, Sergey Ryazantsev } } [SMTP:sryazant-at-ucla.edu] wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FA Q.html } } } -----------------------------------------------------------------------. } } } } } } } } } Randy } } } I am sorry for late comments. I agree with Wil Bigelow that LN2 trap will } } } reduce the pump performance. If you need just to increase the system's } } } overall performance, I would suggest you have to do very good service
for } } } it first (replace all suspicious O-rings, clean everything up). I highly } } } recommend to use Santovac-5 DP (I assume, it's DP based system, because } } TP } } } don't need LN2). When I come in UCLA, I had DV-502A vacuum system with } } } 5*10-5 torr. I cleaned up DP (messy work and a lot of solvents), } } replaced } } } all O-rings (the system was 10 y.o. at the moment) and used } } } Santovac-5. Since that I do have 5*10-7 with no LN2 (and no further } } } service for more than 5 years). As an alternative, you may install some } } } "cold finger" with protective screens near your sample in the Bell } } } Jar. You really need LN2 trap over DP if you pump a lot of water in your } } } experiments. Sergey } } } } } } At 06:09 PM 12/16/02 -0600, you wrote: } } } } ------------------------------------------------------------------------ } } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } } } } Hi All, } } } } I have a Hitachi HUS-4 vacuum evaporator. I'm hoping to put a LN2 } } cold } } } } trap } } } } on it, in an attempt to clean up the vacuum a tad more. There is a 3.5 } } inch } } } } by 4.5 inch plate right above the diffusion stack which looks to be a } } great } } } } place to attach a cold trap. Rumor has it that this was an option. Are } } there } } } } any old traps kicking around that one can acquire? I could have our } } machine } } } } shop manufacture one, but was hoping for a cheaper route. } } } } Thanks in advance, } } } } Randy Nessler } } } } 319-335-8142 } } } } } } ------------------------------------------------------ } } } } } } Sergey Ryazantsev, Ph.D. } } } Electron Microscopy } } } Department of Biological Chemistry, School of Medicine } } } University of California, Los Angeles } } } Box 951737 } } } Los Angeles, CA 90095-1737 } } } } } } (310) 825-1144 (office) } } } Pager: (310) 845-0248 } } } FAX: (310) 206-5272 (departmental) } } } mailto:sryazant-at-ucla.edu } } } } } } } } ------------------------------------------------------ } } Sergey Ryazantsev, Ph.D. } Electron Microscopy } Department of Biological Chemistry, School of Medicine } University of California, Los Angeles } Box 951737 } Los Angeles, CA 90095-1737 } } (310) 825-1144 (office) } Pager: (310) 845-0248 } FAX: (310) 206-5272 (departmental) } mailto:sryazant-at-ucla.edu } } }
Depending on the motivation of the pregnant worker, and the quality of your safety personnel, about a week. In the interim, the person can be given other duties, or in our case just carefully follow the procedures already in place, which have been designed to protect. The biggest question is: Will the lab administration give you the time to get set up before the fact. In a real sense, it is always too late to begin preparations after definitive knowledge of pregnancy is know. In Kathy's case se would have been better off to wait to tell everyone of her condition, this is a shame, however, and we have not come as far as we would like to think with women's rights. In my case it was very beneficial to know that the worker was pregnant, her work dropped off substantially and she napped at the micortome. If I was unaware of the situation. I might have reacted in a negative sense, but instead, I was able to protect her from those who would not understand.
Bill
-----Original Message----- } From: celik ayten [mailto:celik-at-ews.uiuc.edu] Sent: Friday, January 03, 2003 1:20 PM To: William McManus Cc: fisher-at-meganet.net; microscopy-at-sparc5.microscopy.com
On Fri, 3 Jan 2003, William McManus wrote:
[skip]
} 6. Once you know that you have a pregnant worker, contact your safety } people and have the worker prepare a personal hygiene plan and have it } approved by the safety office and suggest that her doctor have a look at } it to see if he/she has any concerns. } I had two pregnant workers in the lab at the same time, there were no } problems. } :
How long is it going to take to go through each step of the procedure mentioned above? By the end of that procedure it might be too late for the fetus.
I believe that as soon as a lady learned or suspect she is pregnant she should stop working with radiation stuff. There is no safe dose of radiation. Risk might be small but it is always positive.
Kind regards, Ayten.
} Just one personal aside, it is much easier to manage an employee if you } know that she is pregnant, some have little to no problems, but others } can have a very hard time. If you do not know what is going on, the } possibility is there to misjudge the performance of that individual, but } the law allows for the worker to privacy. Just one more thing to deal } with in the modern world. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } -----Original Message----- } } From: Cochran [mailto:fisher-at-meganet.net] } Sent: Thursday, January 02, 2003 6:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Pregnancy & SEM } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi All, } } I followed the discussions concerning pregnancy and sem operation with } interest in the past. Unfortunately, I did not save the replies as it } seemed unlikely to occur in our lab. We currently have a pregnant } occassional operator. At her request, the decision has been made to } transfer sem work to another individual for the duration of the } pregnancy. } } When I mentioned that the list server has covered this topic in the } past, I was asked if I the information was still available. I welcome } any new infomation and hope that some one can pass along archived info. } } Thanx, } Ray Cochran } } } }
I've recently developed a problem with a newly purchased Kodak EM film. The EM film box says "new formulation" on it.
Well, this new formulation on film is lowering my contrast and I get some funky fog on it.
I was wondering if anyone else have experienced this problem and if so what can we do?
I've used old film ( old formulation ) and experience no fogging and get nice contrast. Its also not a scope specific problem or light leaks or developer. Its the film.
Rajesh Patel Robert Wood Johnson Medical School Department of Pathology 675 Hoes Lane Piscataway, NJ 08854
Say, can anyone recommend a super-reliable and quiet brand (and size) of air compressor to use with a Philips EM400T?
Unfortunately, in addition to being very, very loud, the low grade air compressor we've been using has the intermittent habit of blowing it's circuit breaker (twice in the last couple of weeks), which has set us way back on getting our "new" microscope fully evacuated for the first time.
I'm waiting for a catalog from Jun-Air, but would like to know what model/size has been used reliably by others over the years. The Philips installation manual doesn't give much more than the air pressure requirement, so I'm uncertain what is ideal in terms of pump and tank capacities. I've asked the folks at Jun-Air, but they are reluctant to make any recommendation based solely on the pressure spec.
Thanks for any tips!
Best regards, Eric Anderson Southern CT State University Physics Dept. 501 Crescent Street, JE108B New Haven, CT 06515 203-392-6468
Those of you planning to attend the 2003 Microscopy and Microanalysis meeting August 3-7, 2003 in San Antonio meeting
I am soliciting presentations for the following session at this upcoming meeting. Related topics described below, as well as outreach programs for grades 6 and above, would be considered. Please contact me should you wish to participate.
See you in Texas!
41. Teaching Microscopy and Imaging in the Digital Age
Organizer: Steve Barlow
Computer technology is changing the ways we access equipment, view samples, and record or manage images. Computer-controlled microscopes and digital imaging have created many new training methods. In addition, these changes make it possible for classroom students to analyze microscopy data and images without requiring access to expensive microscopes. This session will look at new training approaches, such as virtual microscopes and telemicroscopy, and new teaching tools, such as CDs, DVDs, and computer software. The session will discuss using digital technology to create laboratory training protocols and to teach classroom analysis of data contained in digital images. -- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
Rajesh If it's film, nothing to do with it! May be for "new formulation" they suggested "new" developing procedure? Check Instruction and definitely contact Kodak. If this product going under "old" trade name, like "4489 film" it should be exact the same every time. I would suggest, you may return the whole batch of the "new formulation" film back to manufacturer. Kodak sold film through distributors, so it's possible that film was stored improperly before shipped to you (another reason to return it back). Sergey
At 10:53 AM 1/6/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Dear colleague From time to time the issue of EM safety appeared on this ListServer. Somehow, the biggest concern was the safety of the (already) pregnant women. In my point of view, when women IS pregnant and fetus is already developed, IT'S TOO LATE TO TALK (or do something) about safety in the EM Lab! Most our "dangerous" stuff has tendency to be accumulated in the body over time (UA, Pb and so on). Some substances are carcinogens (epoxy resins for instance) and therefore has "long-term" effect to us as well. Breathing the oil mist from rotary pumps (my biggest concern) may cause lung cancer many years later... So, radiation in this '"row" of "dead substances" is nearly negligible - it's easy to detect (how you could detect epoxy on your skin or U in the bones?) and most institutions already has taught regulations on radiation issue. Safety in EM Lab should complain with all local and federal regulations ANY time, not only if pregnant women decided to do EM.
If exposure to the dangerous substances (not only in EM Lab) is a concern, the couple (both male&female) should avoid danger well before they decided to have baby. I would suggest at LEAST 6 month before fertilization they should avoid driving car on freeways (carbon dioxide, poly-carbons, nitric oxide), do not stay close to the buildings with granite decorative panels (some banks has luxury to use granite for ex/in-teriors) - radioactivity!; do not fly (radioactivity again), do not eat fast-food ( carcinogens in the overheated oil). This "poor" couple should also seriously inspect their living place. If place is 20-30 years old, it's very possible that painting contains Pb, some flooring - asbestos (lung cancer) and the wood impregnated with poly-phenols (carcinogen). Who knows, may be 10 years ago somebody broken body-thermometer with mercury and mercury is still here (may cause mental disorders). Synthetic carpet delivers static electricity which may interfere with fertilization (sperm may be shocked by high-voltage discharge!!!). If this hypothetical couple will manage to avoid all those dangerous things, THEN we should start worrying about safety at the working place: computer CRT monitors (radiation), dust, clearness of the drinking water in the fountains/bottled (very questionable), artificial illumination (baby needs natural light to be correctly developed), cubicles (claustrophobia, bad for developed fetus), equipment's noise (may scary the fetus, I am serious), THEN - exposure to the chemicals from neighbor's Lab (somebody spoil acetic acid on the floor) and so on. Each chemical/instrument coming in our Labs has detailed Instruction how to use it. You just has to follow instructions and comply with local and federal regulations.
So, the bottom line is: work in the EM Lab should be SAFE any time and to EVERY person at the level of the local and federal regulations. Each professions has some 'professional risk', which we could not avoid. Signing working contract (or accepting monthly salary payment) we automatically agree with some risk, connected to the duties you suppose to perform. Another issue here, that institution offering the job should clearly state what the professional risk is for this particular job and if for some reasons you don't want to take this risk, you have to quick.
People who seriously concern about pollution should go to Russia: because of economic problems, most industrial manufacture is shout down or working at 5-10% of full power. Russia also has much less cars and widely uses hydro-powered electricity generation (not good for eco-systems, but pollution-free). Russia was only the country at Kioto Conference who DECREASED air pollution over last 10 years. Another advantage being in Russia (I really miss it), the ticket in Bolshoi is $5 and most museums available for free (or 5-10 cents ticket).
Sergey
At 07:22 AM 1/6/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I have followed your submissions to this thread with great interest. It's a new terrain for me. I have a related question.
What advice do you have for a teacher whose pregnant grad student has on-going electron microscopy (SEM/TEM) research project? I would lik to read you views on this.
Many thanks in advance.
Ike Oguocha
--- William McManus {billemac-at-biology.usu.edu} wrote: } } Depending on the motivation of the pregnant worker, and the quality } of } your safety personnel, about a week. In the interim, the person can } be } given other duties, or in our case just carefully follow the } procedures } already in place, which have been designed to protect. The biggest } question is: Will the lab administration give you the time to get set } up } before the fact. In a real sense, it is always too late to begin } preparations after definitive knowledge of pregnancy is know. In } Kathy's case se would have been better off to wait to tell everyone } of } her condition, this is a shame, however, and we have not come as far } as } we would like to think with women's rights. In my case it was very } beneficial to know that the worker was pregnant, her work dropped off } substantially and she napped at the micortome. If I was unaware of } the } situation. I might have reacted in a negative sense, but instead, I } was } able to protect her from those who would not understand. } } Bill
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I have followed your submissions to this thread with great interest. It's a new terrain for me. I have a related question.
What advice do you have for a teacher whose pregnant grad student has on-going electron microscopy (SEM/TEM) research project? I would lik to read you views on this.
Many thanks in advance.
Ike Oguocha
--- William McManus {billemac-at-biology.usu.edu} wrote: } } Depending on the motivation of the pregnant worker, and the quality } of } your safety personnel, about a week. In the interim, the person can } be } given other duties, or in our case just carefully follow the } procedures } already in place, which have been designed to protect. The biggest } question is: Will the lab administration give you the time to get set } up } before the fact. In a real sense, it is always too late to begin } preparations after definitive knowledge of pregnancy is know. In } Kathy's case se would have been better off to wait to tell everyone } of } her condition, this is a shame, however, and we have not come as far } as } we would like to think with women's rights. In my case it was very } beneficial to know that the worker was pregnant, her work dropped off } substantially and she napped at the micortome. If I was unaware of } the } situation. I might have reacted in a negative sense, but instead, I } was } able to protect her from those who would not understand. } } Bill
__________________________________________________ Do You Yahoo!? Everything you'll ever need on one web page from News and Sport to Email and Music Charts http://uk.my.yahoo.com
Invitacion para integrar el Grupo de Trabajo de Telemedicina del Plan Organizacional y Estratégico para la Sociedad de la Información en Argentina
Durante el mes de Diciembre, comienzó la actividad de los Grupos de Trabajo (GT) para la elaboración colectiva de un plan estratégico para el desarrollo de la Sociedad de la Información en Argentina (POESIA). El objetivo fundamental del plan es establecer bases que permitan contribuir a la generación y puesta en marcha de propuestas concretas que contribuyan a la transformación estructural que la Argentina requiere para salir de la crisis.
Se crearon cuatro grupos de trabajos en las áreas elegidas; e-gobierno, e-educación, e-salud (telemedicina) y e-economía. Como parte de las actividades a realizarse en forma on line y en forma presencial, todos los participantes podrán presentar los temas principales que segun su visión, constituirán los ejes tematicos de cada grupo de trabajo. Posterior debate y puesta en comun, los primeros resultados se darán a conocer en un evento presencial a realizarse en abril de 2003 con la presencia de todos los miembros participantes de los GTŽs.
Quienes deseen ampliar la información sobre los requerimientos y mecanismos de participación en el POESIA pueden consultar el sitio web de la nueva asociación (http://www.links.org.ar), en donde podrán solicitar a cada coordinador del GT su inclusión en el grupo de su interés e interiorizarse sobre la metodología de trabajo.
Sobre Links LINKS, la primera Asociación para el Estudio y Promoción de la Sociedad de la Informacion en el pais, fue creada en octubre de 2002. Esta entidad cuenta como socios fundadores a distinguidos investigadores, intelectuales, técnicos y organizadores sociales, especializados en estudios y acciones sobre la Sociedad Informacional, que unen sus esfuerzos con el fin de contribuir a la construcción de una sociedad innovadora, usando las herramientas de las tecnologías de información y comunicación (TIC). Entre los objetivos de la entidad se destacan el estudio y la promoción de la Sociedad de la Información, las acciones para integrar a la población a las nuevas oportunidades, y la transferencia de los conocimientos de la Sociedad Informacional a la comunidad. LINKS pretende constituirse en referente en temas concernientes a la aplicación de una adecuada política de Estado en materia de la Sociedad de la Información tanto en Argentina como en América Latina
Contacto Dra. Lucia E. Muñiz lmuniz-at-links.org.ar
I can remember a couple of messages Robert Grassucci has mailed to the 3D-EM newsgroup in October 2001 and March 2002 concerning a new emulsion for Kodak SO 136 EM-film. He reported a decrease in sensitivity of app. 10% for the new emulsion. He also mentioned reports about an increase in the fog level. To access the original messages you can visit the 3dem messages archive: http://3dem.sdsc.edu/list/maillist.html
Matthias Floetenmeyer
Centre for Microscopy & Microanalysis University of Queensland, Queensland 4072, Brisbane, Australia
I bought a Jun-Air unit (oil type, not oil-less) for use as a dry air source for an FTIR (consumption rate may be higher than for an EM) and found it very tricky. The unit has worked well but the specs are very misleading and they charge extra for every little nut and bolt beyond the basic unit.
If you are using the air only to operate valves, consumption rate is probably not a problem. However if you need a steady stream of air, be aware that the rated flow volume is cited while running full-out.... but the compressor cannot run at more than a 50% duty cycle - so you have to halve the delivery volume right off the bat. Next, the compressor has very small cooling fins and gets excessively hot even on a 50% duty cycle in an ambient of 60 degrees Fahrenheit. When the intake air is preheated (expanded) on the way in, the flow volume drops off very steeply (and the duty cycle becomes excessive and ... there you go again). Therefore you have to have forced cooling. Although it has an overtemp cutout, that apparently applies only to the electrical motor because the compressor head is not supposed to exceed 55 Celsius. Before the fan, I discovered it was running too hot to touch, I would guess about 80 Celsius. They sell an add-on fan but by then my wallet was tapped out and I added my own. The oil darkens a lot in use but so far seems to be functioning properly. I needed two oil mist separators (.5 micron last). Those do seem to function well. The unit is quiet, as described, but draws down the line voltage sharply when it kicks in, even though it is well below the rating for the circuit that it's on. For some reason the starting capacitors are not properly selected for the motor, I guess.
In summary, I read all the specs and went to the local distributor expecting to spend $1,400. I ended up spending $3,000; timing the cycle with a stop watch, and rigging a fan.
John Twilley
Eric Anderson wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } New Year's Greetings! } } Say, can anyone recommend a super-reliable and quiet brand (and size) of } air compressor to use with a Philips EM400T? } } Unfortunately, in addition to being very, very loud, the low grade air } compressor we've been using has the intermittent habit of blowing it's } circuit breaker (twice in the last couple of weeks), which has set us } way back on getting our "new" microscope fully evacuated for the first } time. } } I'm waiting for a catalog from Jun-Air, but would like to know what } model/size has been used reliably by others over the years. The Philips } installation manual doesn't give much more than the air pressure } requirement, so I'm uncertain what is ideal in terms of pump and tank } capacities. I've asked the folks at Jun-Air, but they are reluctant to } make any recommendation based solely on the pressure spec. } } Thanks for any tips! } } Best regards, } Eric Anderson } Southern CT State University Physics Dept. } 501 Crescent Street, JE108B } New Haven, CT 06515 } 203-392-6468
if you use a safelight have you done a check on this with the new film? I usually slightly pre-fog test film (with a light or better still about 1/4 normal e.m. exposure) and then place something on the film a bit nearer to the safelight for several minutes. If you get any shadow of the object, there is a problem. This may require a change of distance, filter or bulb.
The only other obvious problems I can think of is if the developer has gone off or is the wrong one for the new film.
The drop in contrast may just be related to the fogging.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: Rajesh Patel {rpatel-at-umdnj.edu}
Rajesh, I did have a box of film (SO163) just before Christmas which gave very poor results. I suspect that the notch was in the wrong place, which meant that it was loaded emulsion side down in the microscope. What is the accelerating voltage of your microscope? If it's 200 kV or more I guess this would be less noticeable than the blank plates I had. I've never seen this happen before & I guess I've processed 20,000 TEM negatives or so in my career. I imagine film quality control is becoming poorer, since the volume is decreasing due to the rise of digital image capture systems - just as the quality of records went off in the later days of vinyl as CDs became dominant.
Richard _______________________________ Richard Beanland Analytical Services Bookham Technology plc Caswell, Towcester, Northamptonshire NN12 8EQ UK Tel: +44 (0) 1327 356362 Fax: +44 (0) 1327 356775 http://www.bookham.com
-----Original Message----- } From: Rajesh Patel [mailto:rpatel-at-umdnj.edu] Sent: 06 January 2003 18:53 To: Microscopy-at-sparc5.microscopy.com
I've recently developed a problem with a newly purchased Kodak EM film. The EM film box says "new formulation" on it.
Well, this new formulation on film is lowering my contrast and I get some funky fog on it.
I was wondering if anyone else have experienced this problem and if so what can we do?
I've used old film ( old formulation ) and experience no fogging and get nice contrast. Its also not a scope specific problem or light leaks or developer. Its the film.
Rajesh Patel Robert Wood Johnson Medical School Department of Pathology 675 Hoes Lane Piscataway, NJ 08854
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The next major EPSRC FIB meeting is on 4th April 2003 in Cambridge, UK. This will take place directly after Microscopy of Semiconductors MSM XIII.
NanoFIB 2003 will provide an exciting overview of active FIB research both in the UK and abroad. Registration and abstract submission will be via the RMS (www.rms.org.uk).
The website for the new EPSRC NanoFIB network is now running at : www.nanofib.org. Please look for info on FIB meetings, Network members, links to FIB websites. The website is to link together people active in FIB. If you would like something added to the website, such as links to your own websites, send us your suggestions!
Dr. B.J. Inkson Dept. of Engineering Materials The University of Sheffield Sir Robert Hadfield Building, Mappin Street, Sheffield, S1 3JD Tel: 0114 222 5925 FAX: 0114 222 5943 www.nanofib.org
------------------------------------------------- This mail sent through IMP: http://horde.org/imp/
} } I've recently developed a problem with a newly purchased Kodak EM film. The } EM film box says "new formulation" on it. } } Well, this new formulation on film is lowering my contrast and I get some } funky fog on it. } } I was wondering if anyone else have experienced this problem and if so what } can we do? } } I've used old film ( old formulation ) and experience no fogging and get } nice contrast. Its also not a scope specific problem or light leaks or } developer. Its the film. *************************** Rajesh, I have not personally experienced thisa, since I'm still working off an old supply of film, but another EM lab here has been going nuts for the past few months with this same issue. I can't offer any advice, but I will print out & pass along your posting to them. There is comfort in not being alone. Hopefully, we as a large group of interested users can find a solution to this problem, or push Kodak to solve it. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Hi, I would just like to put in my 2 cents worth here. I have been "doing" electron microscopy for over 25 years. I am the mother of a healthy, normal 10 year old boy. He was the end result of a few years worth of false starts. Of course I can never know if the 2 miscarriages were genetic or the result of environmental influences, they were early (6 & 12 weeks) and my vote is with something genetic....I was no spring chicken at the time! During the 3+ years that my quest for motherhood took, I made extra sure that I followed good lab safety procedures. I did double glove when working with fixatives, etc., had the fume hood in my lab inspected and avoided acrylic resins (personal bias?). My 'scope is a 100CX-II that was installed in 1982. It has always been maintained by JEOL and I had no concerns about radiation leaks. I agree with the idea that good lab safety is good lab safety, and if rules are set to protect the most delicate among us, the unborn, and everyone follows them at all times, there should not be a concern. Napping at the microtome, or putting one's feet up when seated at a desk may raise some eyebrows, but it does help you get through the day! Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
The Analytical Imaging Facility of the Albert Einstein College of Medicine has an opening for an experienced electron microscopy technician. For details please see the posting on the MSA Placement Office Job Listings page.
http://www.msa.microscopy.com/PlacementOffice/JobListings.html **************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
Listers; We have been microtoming samples of high impact polystyrene (HIPS) that contains butadiene rubber moieties. These samples have been treated as follows prior to sectioning: 1) Trim block face to prescribed shape and size 2) Harden & stain block by immersing in 2% OsO4 overnight.
Most of the blocks will section fine. However, there is a fair amount of compression of the resulting sections. The sections expand slightly when exposed to chloroform vapors and a lot when exposed to xylene vapors. Sometimes it is necessary to use both to get the sections to expand. However, we are concerned about over expansion and possible swelling of components in the sample. Is there another solvent that could be used to relax the sections without concern for over expansion or swelling of components?
Also, is there someone who does cryo-sectioning of HIPS or similar materials that would be able to do this as a service project for an outside group.
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 West Lafayette, IN 47907
On Monday, January 6, 2003, at 12:41 PM, Eric Anderson wrote:
Dear Eric,
} Say, can anyone recommend a super-reliable and quiet brand (and size) } of } air compressor to use with a Philips EM400T?
Since the FEI people are here installing our microscope, I was able to ask about this. The Jun-Air model 6-25 is the one used both on the new scopes and the older 400 series. } } Thanks for any tips! } You're welcome. Yours, Bill Tivol 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
On Monday, January 6, 2003, at 10:53 AM, Rajesh Patel wrote:
Dear Rajesh,
} I've recently developed a problem with a newly purchased Kodak EM } film. The } EM film box says "new formulation" on it.
Is this SO163? Our newly-purchased SO163 does not say this, but it would be good to know that the new formulation is coming. } } Well, this new formulation on film is lowering my contrast and I get } some } funky fog on it. } } I was wondering if anyone else have experienced this problem and if so } what } can we do? } } I've used old film ( old formulation ) and experience no fogging and } get } nice contrast. Its also not a scope specific problem or light leaks or } developer. Its the film. } Have you tried developing the film in complete darkness? Very sensitive EM films can be fogged by safelights, and sometimes the filter on the safelight can crack and let shorter wavelengths out. Yours, Bill Tivol 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
Dear Richard, I had a run of about 6 to 8 boxes of Kodak 4489 film with the notch on the wrong side, three years ago. I noticed because the emulsion side of the film is lighter under the safelight than the back side, so I loaded them right side up. I sent in the customer reply card, but never heard anything.
} Rajesh, } I did have a box of film (SO163) just before Christmas which gave } very poor results. I suspect that the notch was in the wrong place, which } meant that it was loaded emulsion side down in the microscope. What is the } accelerating voltage of your microscope? If it's 200 kV or more I guess } this would be less noticeable than the blank plates I had. } I've never seen this happen before & I guess I've processed 20,000 TEM } negatives or so in my career. I imagine film quality control is becoming } poorer, since the volume is decreasing due to the rise of digital image } capture systems - just as the quality of records went off in the later days } of vinyl as CDs became dominant. } } Richard } _______________________________ } Richard Beanland
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
Debby Sherman wrote: ============================================================== Listers; We have been microtoming samples of high impact polystyrene (HIPS) that contains butadiene rubber moieties. These samples have been treated as follows prior to sectioning: 1) Trim block face to prescribed shape and size 2) Harden & stain block by immersing in 2% OsO4 overnight.
Most of the blocks will section fine. However, there is a fair amount of compression of the resulting sections. The sections expand slightly when exposed to chloroform vapors and a lot when exposed to xylene vapors. Sometimes it is necessary to use both to get the sections to expand. However , we are concerned about over expansion and possible swelling of components in the sample. Is there another solvent that could be used to relax the sections without concern for over expansion or swelling of components?
Also, is there someone who does cryo-sectioning of HIPS or similar materials that would be able to do this as a service project for an outside group. ================================================================== The laboratory of Structure Probe, Inc. which is the "parent" of SPI Supplies, has been doing cryo sectioning of HIPS for industry for more than thirty years. You are quite correct to be concerned about compression effects in these kinds of samples, because they can create the illusion of orientation when in fact such apparent "orientation" could be an artifact.
There could be many reasons for the compression effects, which could include such factors as the diamond knife itself, the conditions of the ultramicrotomy, temperature of the sectioning ,etc.
Please contact me off-line and describe what the client is trying to accomplish, because that becomes important in terms of determing what orientation the sections should have relative to the injection direction of the molded piece. We can quote you a fixed price to do the entire job. I can not recall any HIPS sample in recent years that could not be treated as a "routine" sample, even those that have been modified with other polymers.
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 ######################## ============================================
IOWA STATE UNIVERSITY An Equal Opportunity/Affirmative Action Employer Vacancy Announcement RESEARCH ASSOCIATE II Vacancy Number: 023649 Department: BIOTECHNOLOGY Proposed Start Date: MARCH, 2003 Appointment Conditions: Continuous, 12 Months, Full Time
Special Conditions: Description: This individual will be responsible for carrying out a variety of laboratory techniques and procedures for the preparation of biological specimens leading to their microscopic observations and analyses for scientists at Iowa State University, and for off-campus scientists/clients. This individual will be responsible for assisting both the Assistant Scientist III and Director of Bessey Microscopy Facility (BMF) in maintaining the laboratory and work environment for all of the researchers who use it. Individual will need to effectively work with researchers and develop a work schedule that fits needs of BMF. Required Qualifications: Bachelor's degree in a biological sciences discipline and one year of laboratory experience that demonstrates knowledge in general and quantitative chemistry, physics, operation and maintenance of light and electron microscopes, ancillary equipment, and all phases of handling chemicals for preparing specimens. Preferred Qualifications: Master's degree in biological sciences discipline, practical experiences in use of light and electron microscopes, rotary and ultramicrotomes, digital imaging and PC computers and BEMT Certification (MSA). Salary/Wage: $30,128 minimum Application Deadline: To guarantee consideration, application must be received by January 30, 2003. Application Instructions: Send cover letter explaining interest, email address, resume, and the names, professional titles and phone numbers of three references to: Dr. Harry T. Horner, Bessey Microscopy Facility, Room 3A Bessey Hall, Iowa State University, Ames, IA 50011-1020, fax: 515-294-1337, or email: hth-at-iastate.edu.
Tracey M. Pepper Supervisor Bessey Microscopy Facility Iowa State University Ames, IA 50011-1020 p. 515.294.3872 f. 515.294.1337
Rajesh: We use Kodak 4489 EM film. The "new formulation" is definitely different in our hands. Our negatives are too thin when we use our standard microscope exposure and chemical processing methods. I have made an acceptable correction by increasing the development time of the film by about 10%, decreasing the exposure of the paper by about 1/3 stop and shortening the development time of the paper by about 10%. I have made no change in the microscope's exposure setting. We use Kodak Kodabrome II RC F4 paper and process both the 4489 film and the Kodabrome paper in a Colex automatic processor using Kodak Polymax RT chemistry. The same correction scheme should work if you are processing in D19 but the percentages will need adjusting.
I wonder what Kodak has done?
Bob -- C. Robert Bagnell, Jr., Ph.D. Associate Professor 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
I have not used TEM film before, but have used lots of photo film. First off, I would never load film under a safe light--it is not safe. Litho film could be handled in yellow light without problem. If the TEM film is pan film, it seems to me that a safe light is not advised.
The flip side of the film is always an issue. If the TEM film is like traditional film, there is a blueish anti-halation coating. That side is the back side. When the film is fixed, the coating is removed. Check an un-developed piece of film and see which is which. Another clue is which side is grainy looking and which is shiny. Grainy is the emulsion side. Probably no surprise to anyone.
When a new box of 4x5 or 220 roll film was purchased, I always developed/processed an unexposed sheet or roll. This was then measured on a densitometer for baseline fog D value. If the D value was lower than normal, the media could be pre-exposed (fogged) to bring the background up to that of other media. The idea is to get the media at a point where the lazy S D/exposure curve was at a point above the toe. Then, there is a large linear range of exposure before the knee, where saturation occurs.
One might try this on old media and the "new" stuff. This would tell whether the new formula is a little different or radically different.
gary g.
At 11:48 AM 1/7/2003, you wrote:
} Dear Richard, } I had a run of about 6 to 8 boxes of Kodak 4489 film with the notch on the } wrong side, three years ago. I noticed because the emulsion side of the film } is lighter under the safelight than the back side, so I loaded them right } side up. I sent in the customer reply card, but never heard anything. } } } } Rajesh, } } I did have a box of film (SO163) just before Christmas which gave } } very poor results. I suspect that the notch was in the wrong place, which } } meant that it was loaded emulsion side down in the microscope. What is } the } } accelerating voltage of your microscope? If it's 200 kV or more I guess } } this would be less noticeable than the blank plates I had. } } I've never seen this happen before & I guess I've processed 20,000 TEM } } negatives or so in my career. I imagine film quality control is becoming } } poorer, since the volume is decreasing due to the rise of digital image } } capture systems - just as the quality of records went off in the later } days } } of vinyl as CDs became dominant. } } } } Richard } } _______________________________ } } Richard Beanland } } Mary Mager } Electron Microscopist } Metals and Materials Engineering } University of British Columbia } 6350 Stores Road } Vancouver, B.C. V6T 1Z4 } CANADA } tel: 604-822-5648 } e-mail: mager-at-interchange.ubc.ca
David, Thanks for such nice comments. My major point was that the Lab should be safe in all possible ways anytime and for EVERYONE. Another point was, that people should think about baby well before they actually get it. The exposure to radiation/chemicals/etc of male and female BEFORE fertilization may cause serious defects in the fetus later. Actually, male's reproductive organs tends to be more sensitive to radiation and some particular chemicals than female's.
From another hand, employees should clearly understand the health risk factor associated with job. In your particular case, those individuals, who is oversensitive to formaldehyde should quit, because this hypersensitivity is a very bad sign: they could develop unusual allergic reactions with very serious result to health in future. I wish all of us to have nice safe conditions in the Lab. Sergey
At 10:46 AM 1/7/2003, you wrote: } I agree in substance, Just a few quick notes. Your concerns } prior to conception are most valid as well as the points that often the } labs are safer than home. There is only so much we can do, but we should } do what we can. There is/should be no legal way to discriminate based on } risks to fetus. So the lab must be made safe for all. } I would disagree on several points: } Federal, State and local safety laws are made for the "Average, } healthy, normal individuals. The TLV's, for example, are created for } someone in this class. Few if any body falls into this class, especially } someone who has just conceived. The presence of formaldehyde is totally } undetectable in the lab, approximately 2 times less than the TLV, yet } without special precautions, I have individuals sensitive to formaldehyde } who react to the small amounts present with skin reactions, or breathing } problems. We attempt to lower formaldehyde vapors to below the limits of } those using the lab, but they are the most sensitive indicators of the } presence of aldehyde vapors, there aren't tests sensitive enough. Kind } of a catch 22. } Oil from vacuum pumps, compared with other risks is relatively } mild. New oil (I know, I know, the breakdown products) has repeatedly } been checked free from carcinogens. (and we still exhaust to the hood system) } At least from my information, Uranyl Acetate is not a bone seeker } like Strontium is. I was told that the biological half life of soluble } uranium was in the order of 6 weeks. The acute danger to kidneys and } liver from the heavy metal toxicity may be as great as the radioactive } hazard to the body. Yes, don't drink it (and especially don't inhale the } dust, there the damage is greater and the biological half life much } longer) but I agree there are many more hazardous chemicals in a EM } Lab. (Cacodylate scares the living hell out of me every time I need to } use it, and I typically don't let any others use it but do the work for them.) } Although the persons accept the risks (making the assumption that } all managers are as careful as you and I in outlining the risks to the } employees and users and training them to avoid them) the bottom line is: } regardless what we have them sign or tell them, the legal and moral } responsibility to protect the employees and users of the laboratory rests } firmly on the manager and it is the managers responsibility to make the } lab safe for all who would wish to use it, regardless of preexisting } conditions, gender etc. } } At 05:40 PM 1/6/2003 -0800, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Why not do an exposure series on the scope with the new film to get more signal (more illumination) so that you don't have to monkey with both film development and print exposure? What you need is a properly exposed negative so that you can get the most out of your prints. If the problem is truly film formulation (slower film) and not some fogging problem, I¹d go for the increased beam intensity.
This is not the first time Kodak has changed formulations and or film format. It doesn't surprise me. We haven't ordered film lately, but it's useful to have this problem discussed here so the rest of us can beware when we do get a new batch.
Thanks, Sara Miller
On Tue, 7 Jan 2003, Robert Bagnell wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Rajesh: } We use Kodak 4489 EM film. The "new formulation" is definitely } different in our hands. Our negatives are too thin when we use our } standard microscope exposure and chemical processing methods. I have } made an acceptable correction by increasing the development time of } the film by about 10%, decreasing the exposure of the paper by about } 1/3 stop and shortening the development time of the paper by about } 10%. I have made no change in the microscope's exposure setting. We } use Kodak Kodabrome II RC F4 paper and process both the 4489 film and } the Kodabrome paper in a Colex automatic processor using Kodak } Polymax RT chemistry. The same correction scheme should work if you } are processing in D19 but the percentages will need adjusting. } } I wonder what Kodak has done? } } Bob } -- } C. Robert Bagnell, Jr., Ph.D. } Associate Professor } 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 } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Though my issue was not regarding pregnancy, I had my concerns over spending a lot of time being bathed in a strong magnetic field. Some studies suggested correlations to increased health problems, whereas other studies suggest some magnetic fields were actually beneficial.?????
Anyway, we went through the lab with a trifield meter to ensure that no one was being unduly exposed to strong emf. While EMs have high voltage and you sit right next to those power supplies, you could reasonably assume that microscope is more sensitive to emf than your body and the manufacturers provide good shielding for image quality.
Alan Stone ASTON Metallurgical Services
At 09:50 AM 1/7/2003 -0500, you wrote: } microscopy-at-sparc5.microscopy.com
Gary TEM film is blue-sensitive, orthochromatic not panchromatic, and red or brown safelights are generally safe if chosen and used according to the manufacturer's instructions. Whether safelight is safe or not depends on film type, safelight wavelength, intensity, time, development, EM image exposure, etc. If you have any anxieties about safe exposure times better to experiment with these and determine what actually happens. Chris
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 08, 2003 12:30 AM
Hi,
I am in the middle of writing a joint grant proposal for an optical disk jukebox / image archiving solution which should hopefully hold up to ten years microscopy data. This system hopefully will aid in 'traceability' of raw data and the image processed result. I was wondering what other research institutes have in place for image archiving regarding traceability of images.
Techniques of image manipulation that, in the past, could only be done by few, and with great care, are now routine, consequently readily available software has also created ample opportunities to falsify images. Obviously the raw data and the final data to be published should be stored long term, but what about the steps in-between? As most research institutes now are multi-user facilities, the onus of scientific proof should be on the owner of the data, but since students are replaced every three - four years there must also be a burden of proof on the research institute especially if the images are to be reused at a later date.
Should a multi-user microscopy facility demonstrate traceability of data or should we rely on the users?
Steve
Steve Bagley Associate Scientist Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute For Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
Sarah's suggestion is a good one and in fact is what we did first. However, changing the exposure of the film in this case did not improve the result. We got more dense negatives but the contrast remained poor. I've used the old photographer's trick of decreasing exposure (in this case keeping it the same) and increasing development (of the film) to improve contrast. This definitely works for the new 4489 formulation, what ever it is.
Bob -- C. Robert Bagnell, Jr., Ph.D. Associate Professor 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
Just a quick note: if you can wait a bit to buy, I would. Microtek (I'm 98% sure it's Microtek) has just come out with a flatbed scanner that uses ICE technology. This is used in the Nikon and other 35mm slide scanners to remove scratches and dust on the film from the scanned image. It's a hardware+software system, not just software. I don't know anything about this scanner yet, other than that it exists, but it would be worth checking out.
Phil
Hope 2003 is good for you all!!
We are looking at purchasing a photo scanner (Probably an EPSON Perfection 2450 Photo). We will be using it to scan old 35mm slides for new Powerpoint presentations.
Since it will also scan negatives and produce positive immages, I was wondering whether it could scan Electron Microscope negatives as well. This would save a good deal of effort and money in enlarging and printing onto photographic paper.
Has anyone tried this? Any recommendations?
Thanks for your consideration,
Tony Henwood JP, BappSc, GradDipSysAnalys, CT(ASC) Laboratory Manager The Children's Hospital at Westmead, Locked Bag 4001, Westmead, 2145, AUSTRALIA. Tel: (02) 9845 3306 Fax: (02) 9845 3318
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-- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Rajesh: We are having the same problems with the "new" film. We tested the scope, camera, safe light, enlarger, desecator, grid holder, and developing solution. We even tested the film in a new scope at Montclair University and also concluded it is the film. We contacted kodak directly, I think you should do the same.
Omayra Velez Electron Microscopy Specialist Pathology Department Weill Cornell Medical College New York, New York 212-746-6437
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I have some problems to prepare the surface of AlMg3 for surface investigations on the one hand and TEM measurements on the other hand.
TEM is less worse because I currently have an electrolyte based on perchloric acid which lets me get satisfying specimen. Most specimen are ion milled afterwards anyway. If there is a better way (without ion milling to save time ;^)) I nevertheless would be very interested.
The other purpose is some more interesting for me: On the one hand I need to investigate the grain structure of my specimen after cold rolling and donŽt get proper grain boundaries to do that. On the other hand I am very interested in proper surfaces for surface observation in DF. I also tried to get a smooth surface with an electrolyte composed of perchloric acid and methanol, but its quite explosive and therefore has to be cooled by solid carbon dioxide...
Does anybody know any other electrolyte or acids to get the results mentioned above?
Thanks in advance,
Hanno Dierke
-- Hanno Dierke, Inst. for Metal Physics an Nuclear Condensed Matter Physics TU Braunschweig, Braunschweig, Germany h.dierke-at-tu-braunschweig.de
I have been working on phase imaging of HIPS with AFM TappingMode for a while since we do not have a TEM here. I tried different microtoming conditions, with a 35-degree diamond knife on a microtone for light microscope samples (minimum 0.5 um sectioning), for the best cutting faces and therefore the imaging from them, but with limited success. I found ca. -30 to -50 C is probably the "right" temperature, but the resulted facing quality varies from sample to sample. Defects are primarily chatters and local compressions on polystyrene matrix. I also found difficulties in parameter settings of the DI AFM for different HIPS samples. However, the same operation is very successful for ICPs (impact copolymer of polypropylene and polyethylene).
Does anybody have any experiences, suggestions or comments? Many thanks in advance.
Debby, did you try AFM on HIPS?
Jiang Liu, PhD Research and Technology Center Atofina Petrochemicals, Inc. Tel: 281-884-0529 email: jiang.liu-at-atofina.com
} From: Debby Sherman {dsherman-at-purdue.edu} } To: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} } Subject: Sectioning HIPS } Date: Tue, 07 Jan 2003 11:35:41 -0500 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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Hi Steve, As the digital revolution has progressed, I have thought image tracability will become a major issue. I have wondered if anyone has developed a method of tagging the original data file in a way that you could always identify the original? So far people have relied on image analysis techniques to determine originals from manipulations and/or actual theft of images or parts of images.
Robert Underwood U of Washington Seattle
On Wed, 8 Jan 2003, Steve Bagley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Hi, } } I am in the middle of writing a joint grant proposal for an optical disk } jukebox / image archiving solution which should hopefully hold up to ten } years microscopy data. This system hopefully will aid in 'traceability' of } raw data and the image processed result. I was wondering what other research } institutes have in place for image archiving regarding traceability of } images. } } Techniques of image manipulation that, in the past, could only be done by } few, and with great care, are now routine, consequently readily available } software has also created ample opportunities to falsify images. Obviously } the raw data and the final data to be published should be stored long term, } but what about the steps in-between? As most research institutes now are } multi-user facilities, the onus of scientific proof should be on the owner } of the data, but since students are replaced every three - four years there } must also be a burden of proof on the research institute especially if the } images are to be reused at a later date. } } Should a multi-user microscopy facility demonstrate traceability of data or } should we rely on the users? } } } } Steve } } } Steve Bagley } Associate Scientist } Applied Imaging Facility } Paterson Institute For Cancer Research } Cancer Research UK } Christie Hospital, Wilmslow Rd, } Manchester. M20 9BX, UK } } } } } } } } } } } } } } } } } } } } } This email is confidential and intended solely for the use of the Person(s) } ('the intended recipient') to whom it was addressed. Any views or opinions } presented are solely those of the author and do not necessarily represent } those of the Paterson Institute For Cancer Research or the Christie Hospital } NHS Trust. It may contain information that is privileged & confidential } within the meaning of applicable law. Accordingly any dissemination, } distribution, copying, or other use of this message, or any of its contents, } by any person other than the intended recipient may constitute a breach of } civil or criminal law and is strictly prohibited. If you are NOT the } intended recipient please contact the sender and dispose of this e-mail as } soon as possible. } }
University of Connecticut Institute of Materials Science
Postdoctoral Position Displacive Phase Transformations
A post-doctoral position is available immediately within the Institute of Materials Science at the University of Connecticut to work on displacive phase transformations in crystalline solids. The systems to be studied include metallic pseudoelastic alloys and ferroelectric thin films. The ideal candidate will have experience in both microstructural characterization (particularly TEM) and phenomenological/crystallographic modeling of phase transformations. This position is a one-year appointment, with possible renewal for a second year.
To apply, please send a complete resume, together with a list of publications and contact details for 3 references to either Prof. S. Pamir Alpay (p.alpay-at-ims.uconn.edu) or Prof. Mark Aindow (m.aindow-at-uconn.edu). Screening of applications will begin immediately, and will continue until the position is filled. We encourage applications from under-represented groups, including minorities, women and people with disabilities.
Mark Aindow, Associate Professor, Department of Metallurgy and Materials Engineering Institute of Materials Science, 97 North Eagleville Road, Unit 3136 University of Connecticut, Storrs, CT 06269-3136, USA
} Tony, } } Just a quick note: if you can wait a bit to buy, I would. Microtek } (I'm 98% sure it's Microtek) has just come out with a flatbed scanner } that uses ICE technology. This is used in the Nikon and other 35mm } slide scanners to remove scratches and dust on the film from the } scanned image. It's a hardware+software system, not just software. } I don't know anything about this scanner yet, other than that it } exists, but it would be worth checking out.
But be aware that any camera or scanner hardware/software that "removes dust and scratches" usually does so by blurring and eroding and then unblurring, or similar processing, to despeckle or hide items identified as dust or scratches, thereby changing image pixels and altering your data. These programs may identify "real" objects in electron micrographs as dust and delete them! The technology is great for restoring that old photograph of your great-grandma, but has the potential to change your scientific data. Try to find out how the hardware/software works before deciding which to buy.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Ann: The new film has the letters "New Formulation" written in white over a red slip, on the side or top of the box. Kodak already contacted us today and we are in process to send them our test material(negatives, prints, and info). If you do not have this film then you are lucky, is been a nut house here, with these unprintable negatives.
Omayra Velez Electron Microscopy Specialist Weill Cornell Medical College New York, New York. 010021 212-746-6437
--- "R. Ann Bliss" {bliss5-at-llnl.gov} wrote: } Omayra: } } After following the thread about the "new" film for } a few days now, I } finally went to check the new shipment of film I } received last week. } Where does one find the words "New Formulation" or } whatever it was? i } looked all over the outside of the box and did not } see those words. } Am I lucky and don't have the problem (yet) or is it } on the paper } work I never read on the inside? } } -- } } +++++++++++++++++++++++++++++ } } R. Ann Bliss } Technician, Chemistry and Materials Science } Materials Science and Technology Division } Lawrence Livermore National Laboratory } } _____________________________
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you are touching on an issue that is probably of interest to a much larger audience. There are at least two areas, where this has been an issue for a while: Forensics and Pharmaceuticals. The Pharmaceutical industry in the US is required by the FDA to keep track of data because lives may depend on it. It used to be lab books that needed to be kept in certain ways, but now that much of the data is electronic, the FDA has published requirements for the safekeeping of data (21 CFR rule 11). It may be interesting for you to see if this answers some of your questions (caution: 21 CFR rule 11 is kind of messy). But even rule 11 relies on SOPs to ensure data safety and security. Traceability and audit features are part of 21 CFR rule 11.
21 CFR rule 11 may be overkill for many settings, and a relatively simple image archive or data base might suffice. Set it up so that certain information must be entered (for example sample number, patient ID, etc.), keep the images on a central server and have all imaging instruments store their data there, and it might do what you want.
I don't think that you can have a 100% watertight system without some decisions by the users, because it would require, that ALL images are stored, regardless of content. And even then, the choice of what images to take still lies with the individuals. You probably have to set up some SOPs that tell the users that they need to save the images within a system that allows some measure of control, be it in an archive or save them to CD-R which then have to be turned in for safekeeping. The question then turns to finding a system that supports the control you are looking for.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Steve Bagley [mailto:SBagley-at-picr.man.ac.uk] Sent: Wednesday, January 08, 2003 5:49 AM To: 'Micro Discussion Group'
Hi,
I am in the middle of writing a joint grant proposal for an optical disk jukebox / image archiving solution which should hopefully hold up to ten years microscopy data. This system hopefully will aid in 'traceability' of raw data and the image processed result. I was wondering what other research institutes have in place for image archiving regarding traceability of images.
Techniques of image manipulation that, in the past, could only be done by few, and with great care, are now routine, consequently readily available software has also created ample opportunities to falsify images. Obviously the raw data and the final data to be published should be stored long term, but what about the steps in-between? As most research institutes now are multi-user facilities, the onus of scientific proof should be on the owner of the data, but since students are replaced every three - four years there must also be a burden of proof on the research institute especially if the images are to be reused at a later date.
Should a multi-user microscopy facility demonstrate traceability of data or should we rely on the users?
Steve
Steve Bagley Associate Scientist Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute For Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
The videotapes from the 2002 MSA tutorials in Quebec City are now available. We are now in a positions to offer the entire library on DVD as well as VHS. We expect to phase out the VHS versions as the current supply that is on hand is exhausted. The most up to date catalog can be viewed at the MSA website or go directly to this URL: http://www.biotech.ufl.edu/sems/newtape00.htm As time permits, I am trying to make abstracts of each of the tapes available online as well so that one has a better idea of what was covered in the tutorial. If you are unable to access this site I can provide a printed copy or I can email a copy of the catalog. We are now also able to accept VISA or Mastercard for payment. We will also accept institutional purchase orders for which we will invoice. As you will notice many of the tapes are quite old and the topics covered certainly deserve being updated. The Education Committee invites interested parties to present tutorials or short courses on these topics, or topics never covered that might be appropriate for tutorials. The committee may be contact at this address MSAEdCommittee-at-MSA.Microscopy.Com
Please feel free to contact me concerning the video library.
Greg Erdos Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
It is the ScanMaker 6800. This scanner will scan reflective media up to 8.5x11" and transmissive media up to 4x5".
http://www.microtekusa.com/indexICEflash.html
It uses Digital ICE, as do the high end Nikon CoolScan scanners. The principle of image correction is based on a separate IR channel. It scans once to pick up greyscale or RGB info, then a second time for IR. Then the image is processed and "fixed" if bad. One web link says two passes while another says only one pass. ?? Anyway, about D-ICE... Sometimes it works really good. Other times not so good. So, case by case basis. I use the Nikon Super CoolScan 8000 for slides and medium format media. It is optical 4K x 4K dpi with D-ICE and GEM.
It (D-ICE) does not work on old Kodachrome transparencies since they block the IR. Other media seem OK. It has a bit of difficulty with b/w slides. The 6800 is 4800x2400 dpi optical, which ought to be OK for most applications. With Firewire and USB for MSRP $399, that is pretty good.
gary g.
At 09:49 AM 1/8/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Greetings, Tina's comment is right on. Note also that the original poster was seeking in part to move their 'good old' projection slides over to powerpoint. Well, if you are like me, over the years you have been showing your favorite slides they have accumulated plenty of dust. If its ok in the slide projector, why worry about it in powerpoint? It will give your presentation a comfy well worn look like an elbow patch on a tweed jacket. You can of course also do nice dodging and burning in photoshop or equivalent to remove egregious dust specks before dropping them into powerpoint.
Just a thought, Tobias
} } } } On Wed, 8 Jan 2003, Philip Oshel wrote: } } } Tony, } } } } Just a quick note: if you can wait a bit to buy, I would. Microtek } } (I'm 98% sure it's Microtek) has just come out with a flatbed scanner } } that uses ICE technology. This is used in the Nikon and other 35mm } } slide scanners to remove scratches and dust on the film from the } } scanned image. It's a hardware+software system, not just software. } } I don't know anything about this scanner yet, other than that it } } exists, but it would be worth checking out. } } But be aware that any camera or scanner hardware/software that "removes } dust and scratches" usually does so by blurring and eroding and then } unblurring, or similar processing, to despeckle or hide items identified } as dust or scratches, thereby changing image pixels and altering your } data. These programs may identify "real" objects in electron micrographs } as dust and delete them! The technology is great for restoring that old } photograph of your great-grandma, but has the potential to change your } scientific data. Try to find out how the hardware/software works before } deciding which to buy. } } Aloha, } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } ****************************************************************************
Many, many thanks for all your thoughts and generous replies!
Since the budget allows it, (at the moment), I'm going to follow the crowd (and FEI technicians) who have suggested the Jun-Air 6-25. (Many wrote just to me and not to the whole group. Jim, unfortunately the folks at Jun-Air would not recommend a model suited to my application when I asked them. They only offered to send a catalog.)
Hopefully, my particular application and environment won't generate the problems John experienced. We really need the quiet operation (specs say 45 dB), especially because the unexpected cycling of a louder compressor is scaring folks out of their wits. Not at all a good situation for handling delicate specimens/procedures.
I'm very tempted to just use nitrogen tanks, but the old 400 has a couple of slow leaks which I might not get to fixing right away, plus we'll probably have it running continuously for long periods, so I'd rather not worry about tank deliveries and running out at a bad time. (One of the same reasons I switched to natural gas from oil for home heating..., plus cleanliness of course.)
Vitaly, thanks for the reasonably priced alternative that employs a modified refrigeration compressor. I love being creative, but just don't have the time right now. Will keep it in mind though.
Thanks & best regards to all! -Eric
} } } Eric Anderson wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } New Year's Greetings! } } } } Say, can anyone recommend a super-reliable and quiet brand (and size) of } } air compressor to use with a Philips EM400T? } } } } Unfortunately, in addition to being very, very loud, the low grade air } } compressor we've been using has the intermittent habit of blowing it's } } circuit breaker (twice in the last couple of weeks), which has set us } } way back on getting our "new" microscope fully evacuated for the first } } time. } } } } I'm waiting for a catalog from Jun-Air, but would like to know what } } model/size has been used reliably by others over the years. The Philips } } installation manual doesn't give much more than the air pressure } } requirement, so I'm uncertain what is ideal in terms of pump and tank } } capacities. I've asked the folks at Jun-Air, but they are reluctant to } } make any recommendation based solely on the pressure spec. } } } } Thanks for any tips! } } } } Best regards, } } Eric Anderson } } Southern CT State University Physics Dept. } } 501 Crescent Street, JE108B } } New Haven, CT 06515 } } 203-392-6468
I have a GATAN Cryo-Transfer cold stage for a Philips EM420 TEM. The crystal needs to be replaced and I was wondering if anyone has knowledge of a third party who does this type of repair?
Thanks in advance.
Tom -- Thomas M. Murray Phone: (206)543-2836 Manager, Electron Microscopy Center Fax: (206)543-3100 Materials Science & Engineering email: murraytm-at-u.washington.edu 302 Roberts Hall, Box 352120 University of Washington Seattle, WA 98195-2120
before you rush out and buy the Scanmaker 6800 for its Digital ICE scratch correction you should be aware that this is only available for prints and not negatives. Although the scanner takes negatives, there is no infra red light source which is required for scratch correction on film - see PC magazine review at: http://www.microtekusa.com/images/pcmag6800.pdf This is pointed to on the Microtek page: http://www.microtekusa.com/indexICEflash.html
I also assume that negatives are scanned on a glass plate rather than a glassless carrier of the Microtek Scanmaker 8700 or 1800f types so you will get more dust marks etc.
Malcolm
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: Gary Gaugler {gary-at-gaugler.com}
Morning Folks,
I am in the process of restarting a lab which has a Zeiss EM900 TEM. When I got here, I found the manual waterlogged! Another flood in an EM Lab! I am in need of an operators and any other electrical and mechanical manuals that pertain to this scope. I am willing to pay copying and shipping costs.
Thanks,
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 calomeni-1-at-medctr.osu.edu
I've run into compressor issues with every Philips scope I've worked with, what has always gotten me though is the fact that reliable quite air compressors are reality not fantasy since JEOL provides air compressors that are just that, and the HVAC systems for most buildings also using 24/7 air compressor systems for the thermostats (the little hiss when you change the thermostat on the wall?) as seem to work well.
Our current system is on a Philips 505. But after having tried several compressors and repairs, I have wound up building my own compressor system out of various components.
(1) Using 20 gallon storage tank from previous compressor
(2) Added a Thomas Compact Oilless Reciprocating Air Compressor ($400). Go well over pressure and volumne needed so as to not stress the compressor. Remeber your really don't need much volumne.
(3) Built a mounting plate out of aluminium and large rubber bushings
(4) Using high quality air hose as vibration isolation and copper tubing.
(5) Using an Ashcroft indusdrial pressure switch for turning compressor on/off ($130). Mounted on the wall off the compressor (vibration its a killer) - NOTE: **Do not** go cheap on this component, trust me! I run the compressor pressure just high enough to minimize cycling and provide the needed pressure at the scope without relying heavily on pressure regulators. Pushing the compressors to their "rated" pressure mark and using a regulator will reduce cycling, but it adds wear to the compressor (Yes, you can run your car just below red line on the tach, but for how long, eh?)
(6) a few oil filled air pressure gauges
(7) Reusing an exisiting air drying system.
(8) Installed the compressor in the service chaseway behind the scope for more noise isolation. (I do recommend getting it out of the scope room if possible, along with the water chillers. Soldered copper piping workes well.)
---} This system has been running (knock-on-wood) very well for over four years now without a problem. Cycle duty is less than 20% even with heavy microscope use. The noise is very low, next to the compressor, and barely noticable in surrounding rooms (don't have a dB meter).
---} Get the air leaks fixed on your 400!
Good luck.
} } Many, many thanks for all your thoughts and generous replies! } } Since the budget allows it, (at the moment), I'm going to follow the crowd (and } FEI technicians) who have suggested the Jun-Air 6-25. (Many wrote just to me and } not to the whole group. Jim, unfortunately the folks at Jun-Air would not } recommend a model suited to my application when I asked them. They only offered } to send a catalog.) } } Hopefully, my particular application and environment won't generate the } problems John experienced. We really need the quiet operation (specs say 45 } dB), especially because the unexpected cycling of a louder compressor is scaring } folks out of their wits. Not at all a good situation for handling delicate } specimens/procedures. } } I'm very tempted to just use nitrogen tanks, but the old 400 has a couple of } slow leaks which I might not get to fixing right away, plus we'll probably have } it running continuously for long periods, so I'd rather not worry about tank } deliveries and running out at a bad time. (One of the same reasons I switched } to natural gas from oil for home heating..., plus cleanliness of course.) } } Vitaly, thanks for the reasonably priced alternative that employs a modified } refrigeration compressor. I love being creative, but just don't have the time } right now. Will keep it in mind though. } } Thanks & best regards to all! } -Eric } } } } } } } Eric Anderson wrote: } } } } } ------------------------------------------------------------------------ The } } } Microscopy ListServer -- Sponsor: The Microscopy Society of America To } } } Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line } } } Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } New Year's Greetings! } } } } } } Say, can anyone recommend a super-reliable and quiet brand (and size) of air } } } compressor to use with a Philips EM400T? } } } } } } Unfortunately, in addition to being very, very loud, the low grade air } } } compressor we've been using has the intermittent habit of blowing it's } } } circuit breaker (twice in the last couple of weeks), which has set us } } } way back on getting our "new" microscope fully evacuated for the first } } } time. } } } } } } I'm waiting for a catalog from Jun-Air, but would like to know what } } } model/size has been used reliably by others over the years. The Philips } } } installation manual doesn't give much more than the air pressure } } } requirement, so I'm uncertain what is ideal in terms of pump and tank } } } capacities. I've asked the folks at Jun-Air, but they are reluctant to make } } } any recommendation based solely on the pressure spec. } } } } } } Thanks for any tips! } } } } } } Best regards, } } } Eric Anderson } } } Southern CT State University Physics Dept. } } } 501 Crescent Street, JE108B } } } New Haven, CT 06515 } } } 203-392-6468 } } }
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
Hello. I am a graduate student at Hofstra University, currently working on a retinal analysis of a deep water shark. I am headed on my first specimen collection trip this week and have a question about my fixation protocol. I will be using a mix of paraformaldehyde/gluteraldehyde in cacodylate buffer at pH 7.2. Should I be using sucrose, CaCl2, etc..to adjust the tonicity?
Any advice would be appreciated. Thank you.
Jill Olin Graduate Student Department of Biology Hofstra University Hempstead, NY 11549
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{color} {param} 0100,0100,0100 {/param} {bigger} Hi everybody,
We are looking into buying a new vacuum evaporator. We would it like to be equipped in two electrodes (for carbon and metal evaporation), rotary/tilt stage and thickness monitor. Our laboratory deals mainly with biological/veterinary samples. I would like to hear your recommendations regarding the purchase: equipment servicing and make (which one is good which is not). Is Denton the best available on the market? Do they still supply DV 502A?
Contact Tom Schmelzer at TGS Technologies. And/or see his ad in the November Microscopy Today on page 44.
He can be reached at 724 453 3865
-----Original Message----- } From: Tom Murray [mailto:murraytm-at-u.washington.edu] Sent: Wednesday, January 08, 2003 7:59 PM To: Microscopy
Hi All,
I have a GATAN Cryo-Transfer cold stage for a Philips EM420 TEM. The crystal needs to be replaced and I was wondering if anyone has knowledge of a third party who does this type of repair?
Thanks in advance.
Tom -- Thomas M. Murray Phone: (206)543-2836 Manager, Electron Microscopy Center Fax: (206)543-3100 Materials Science & Engineering email: murraytm-at-u.washington.edu 302 Roberts Hall, Box 352120 University of Washington Seattle, WA 98195-2120
The consensus seems to be that Tom Schmelzer at TGS Technologies is a good choice.
Thanks for the replies.
Tom -- Thomas M. Murray Phone: (206)543-2836 Manager, Electron Microscopy Center Fax: (206)543-3100 Materials Science & Engineering email: murraytm-at-u.washington.edu 302 Roberts Hall, Box 352120 University of Washington Seattle, WA 98195-2120
The EM Facility at Utah State University is being closed, March 1st 2003 due to budget cuts to the University. I am looking for other facilities which will do service work, and pricing rates for that work. Some of the work will be for University personnel, but most will be for off-site industrial users. Please let me know if you can do work for one group or the other or both. This includes both biological and materials; TEM and SEM (high res, and environmental) and edx.
Thank you for any help in this matter.
Bill
William McManus Supervisor Microscopy Facility Utah State University Logan UT 84322-5305
Tina's comments are quite on the mark. I have a Nikon Coolscan 4000 ED with the scratch removal software (ice cube). I do not use it at all after some tests. The effect is usually too strong and there is no preview of the effect as it is possible in photoshop. So I scan in regular 35 mm slides with no scratch removal, and then take them over to photoshop, use "Dust and Scratches" diameter usually 2 pixel, threshold 8 to 12, and check on some areas with particular detail. Occasionally even that is too much, and then it is back to the cloning stamp and manual removal of dust and scratches. Use a feathered cloning stamp: I like diameter 9 - 13 pixels and 75% hardness, 5% spacing. The feathered tool avoids hard edges around the cloned areas. Takes a while on a 55-60 MB file (RGB TIFF). Note, that with the fine removal of dust, you may not be able to get the larger pieces, which still have to be removed manually. Also, take some time in prepping your hard copy images: use a dust removal spray, and possibly a fine optical lens brush. The 10 seconds spent on that are well worth the time not spent messing around in photoshop.
I would think that the scratch removal effects will be much more disastrous on TEM images, in which the images is directly lens formed (i.e., not pixelated), than with SEM images. In the latter, on good old 4 x 5" negatives you get glorified pixels on a fine grain emulsion, so scratch removal should be quite effective on SEM negatives, but may pose problems with TEM images.
Best wishes Daniel
At 7:49 AM -1000 1/8/03, Tina Carvalho wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} But be aware that any camera or scanner hardware/software that "removes } dust and scratches" usually does so by blurring and eroding and then } unblurring, or similar processing, ...
Not exactly true ... as mentioned by another post, if this scanner uses the ICE procedure, then it uses an infrared scan to detect dust and thereby corrects only the areas within the image which needs it.
What needs to be mentioned in addition is the fact IR cannot see through a silver emulsion, and will therefore see all exposed areas as needing correction. IR aided correction will only work with color transparencies and chromes.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
We do all manner of biological TEM and would be happy to help out if the need arises.
Sara Miller Contact info in footer
On Thu, 9 Jan 2003, William McManus wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear group: } } The EM Facility at Utah State University is being closed, March 1st 2003 } due to budget cuts to the University. I am looking for other facilities } which will do service work, and pricing rates for that work. Some of } the work will be for University personnel, but most will be for off-site } industrial users. Please let me know if you can do work for one group } or the other or both. This includes both biological and materials; TEM } and SEM (high res, and environmental) and edx. } } Thank you for any help in this matter. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Our industry is currently implementing systems to deal with regulatory agency requirements as stated by Mike Bode in his recent reply. There are a few companies working on database solutions to solve some of the traceability issues related to electronic raw data and data manipulation. I am aware of 2 companies that are marketing database applications to address these issues (Cyberlab / Nugenesis(SP?)), and there are probably many more companies. Keep in mind that these applications are not specifically targeted at imaging data, but rather will address most file types. In short, the raw data is sent to a server, the database then archives the file to the database (at some user specified interval). To view the information the files are "restored" to a specified location where they can be manipulated. These manipulated files are then returned to the database using version control methodologies. An audit trail is maintained which allows for complete tracking of raw data access and manipulations. In our installation the archived files are maintained on servers making raw data retrieval a simple affair i.e. no CD's or tapes need be changed / loaded.
In my opinion this type of archival system would be very cumbersome and prohibitively expensive for most EM facilities to implement AND maintain.
One final note: It's a sad state of affairs when the assumption is made that these systems are required to prevent fraud. Dishonest people will always find a way to "beat" the system. It was true for wet chemistry methods and it is true for digital imaging as well. In my mind these systems and procedures are required to prevent the inadvertent, unknowing alteration of raw data. Accidentally deleting or overwriting a file, "misplacing" a file, or possibly confusing a raw data file with one that you knowingly manipulated during analysis. I am speculating but I would assume that most of us have experienced some or possibly all of these situations at one point in our digital careers. Honest mistakes, yes, but troubling errors that must be avoided. They waste time, effort, money, and can be very costly in terms of your reputation and the credibility of your work.
My two cents worth.
John Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
Phone (203)798-5640 Fax (203)798-5698
e-mail jrobson-at-RDG.boehringer-ingelheim.com
-----Original Message----- } From: Steve Bagley [mailto:SBagley-at-picr.man.ac.uk] Sent: Wednesday, January 08, 2003 7:49 AM To: 'Micro Discussion Group'
Hi,
I am in the middle of writing a joint grant proposal for an optical disk jukebox / image archiving solution which should hopefully hold up to ten years microscopy data. This system hopefully will aid in 'traceability' of raw data and the image processed result. I was wondering what other research institutes have in place for image archiving regarding traceability of images.
Techniques of image manipulation that, in the past, could only be done by few, and with great care, are now routine, consequently readily available software has also created ample opportunities to falsify images. Obviously the raw data and the final data to be published should be stored long term, but what about the steps in-between? As most research institutes now are multi-user facilities, the onus of scientific proof should be on the owner of the data, but since students are replaced every three - four years there must also be a burden of proof on the research institute especially if the images are to be reused at a later date.
Should a multi-user microscopy facility demonstrate traceability of data or should we rely on the users?
Steve
Steve Bagley Associate Scientist Applied Imaging Facility Paterson Institute For Cancer Research Cancer Research UK Christie Hospital, Wilmslow Rd, Manchester. M20 9BX, UK
This email is confidential and intended solely for the use of the Person(s) ('the intended recipient') to whom it was addressed. Any views or opinions presented are solely those of the author and do not necessarily represent those of the Paterson Institute For Cancer Research or the Christie Hospital NHS Trust. It may contain information that is privileged & confidential within the meaning of applicable law. Accordingly any dissemination, distribution, copying, or other use of this message, or any of its contents, by any person other than the intended recipient may constitute a breach of civil or criminal law and is strictly prohibited. If you are NOT the intended recipient please contact the sender and dispose of this e-mail as soon as possible.
We are going to process a biofilm with benign strain of E. coli on surface of catheters for TEM. We are reluctant to scrape the biofilm off, so we are going to process the biofilm while attached to the catheter. We do not know if the biofilm, catheter and resin (Glauert Med Embed) will embed well and the sections will come out good. We appreciate any suggestions before we try this out.
Thank you,
Claire Haueter
Claire Haueter EM Technician Integrated Microscopy Core Baylor College of Medicine 713-798-4952
I am a graduate student with the University of Auckland requiring advice on (1) the post-fixation stage for TEM prep., (2) any advice concerning the treatment of fragile sample material for TEM (standard viewing or shadowing).
I am using TEM to look at freshly grown microstromatolitic films from a geothermal brine where the primary mineral component is colloidal silica, with a minor component of sulphur. The objective is to study biomineralisation. We have scrape one set of my samples from their glass substrate (a terribly destructive stage as far a texture goes as the samples are very brittle and porous), dehydrated them through an ethanol series and added osmium tetroxide as a post-fixative (after initial fixation with 2.5% glutaraldehyde/buffer), however in similar studies the osmium tetroxide stage has been omitted - any ideas why?
I would be greatful for any advice.
Kind Regards, Kim -- Kim M. Handley Geology Department University of Auckland k.handley-at-auckland.ac.nz
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} Steve:
There was symposium on this very topic at the MSA meeting in Quebec City this past summer. I taped most of it and when I find the time (ha!) the plan is to publish a synopsis in Microscopy Today and on the web.
The bottom line if I recall correctly, is that current Copyright Law currently provides most - but not all - protection....at least in the US. Probably there will be variations country-to-country though.
Peter Ingram Public Policy Chair and Legal Liaison Officer, MSA
} } } } } } } } } } } } } } } } } } This email is confidential and intended solely for the use of the Person(s) } ('the intended recipient') to whom it was addressed. Any views or opinions } presented are solely those of the author and do not necessarily represent } those of the Paterson Institute For Cancer Research or the Christie Hospital } NHS Trust. It may contain information that is privileged & confidential } within the meaning of applicable law. Accordingly any dissemination, } distribution, copying, or other use of this message, or any of its contents, } by any person other than the intended recipient may constitute a breach of } civil or criminal law and is strictly prohibited. If you are NOT the } intended recipient please contact the sender and dispose of this e-mail as } soon as possible.
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
We are going to process a biofilm with benign strain of E. coli on surface of catheters for TEM. We are reluctant to scrape the biofilm off, so we are going to process the biofilm while attached to the catheter. We do not know if the biofilm, catheter and resin (Glauert Med Embed) will embed well and the sections will come out good. We appreciate any suggestions before we try this out.
Thank you,
Claire Haueter
Claire Haueter EM Technician Integrated Microscopy Core Baylor College of Medicine 713-798-4952
Briefly, you can set the resin in a section of catheter. Then peel off the catheter and reembed your core in a suitable mold. A lot of the biofilm should remain in the resin.
Dave
On Thu, 9 Jan 2003 20:22:49 -0600 chaueter {chaueter-at-bcm.tmc.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello Listers, } } We are going to process a biofilm with benign strain of E. coli on surface of } catheters for TEM. We are reluctant to scrape the biofilm off, so we are } going to process the biofilm while attached to the catheter. We do not know } if the biofilm, catheter and resin (Glauert Med Embed) will embed well and the } sections will come out good. We appreciate any suggestions before we try this } out. } } Thank you, } } Claire Haueter } } Claire Haueter } EM Technician } Integrated Microscopy Core } Baylor College of Medicine } 713-798-4952 } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
As for your evaporator enquiry on the best evaporator, we have reason to believe Ladd Research provides the best vacuum evaporator available and has for many years.
Naturally I must confess to a little bias on the subject, but not too much. You can check our website for more information or contact us.
Disclaimer: Ladd sells all microscopy supplies and accessories, including the Ladd Vacuum Evaporator.
Best regards, John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- From: Dorota Wadowska To: microscopy-at-sparc5.microscopy.com Sent: Thursday, January 09, 2003 8:10 AM Subject: vacuum evaporator
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html ---------------- -------------------------------------------------------.
Hi everybody, We are looking into buying a new vacuum evaporator. We would it like to be equipped in two electrodes (for carbon and metal evaporation), rotary/tilt stage and thickness monitor. Our laboratory deals mainly with biological/veterinary samples. I would like to hear your recommendations regarding the purchase: equipment servicing and make (which one is good which is not). Is Denton the best available on the market? Do they still supply DV 502A? Thanks Dorota
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
Try to avoid disturbing the biofilm if yu substatum is cheap by adding resin etc to it. People have done this with cell cultures on coverslips. The resin and mold can be separated from the glass using liquid nitrogen. The biofilm should stay with the resin.
Dave
On Fri, 10 Jan 2003 16:18:33 +1300 Kim Handley {khan030-at-ec.auckland.ac.nz} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } } I am a graduate student with the University of Auckland requiring advice on (1) } the post-fixation stage for TEM prep., (2) any advice concerning the treatment } of fragile sample material for TEM (standard viewing or shadowing). } } I am using TEM to look at freshly grown microstromatolitic films from a } geothermal brine where the primary mineral component is colloidal silica, with } a minor component of sulphur. The objective is to study biomineralisation. } We have scrape one set of my samples from their glass substrate (a terribly } destructive stage as far a texture goes as the samples are very brittle and } porous), dehydrated them through an ethanol series and added osmium tetroxide } as a post-fixative (after initial fixation with 2.5% glutaraldehyde/buffer), } however in similar studies the osmium tetroxide stage has been omitted - any } ideas why? } } I would be greatful for any advice. } } Kind Regards, } Kim } -- } Kim M. Handley } Geology Department } University of Auckland } k.handley-at-auckland.ac.nz } } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Hi all A happy new year to all. May this year be the year your lottery numbers come up!
Anaspec is a EM Technical support company operating from South Africa but supporting clients around the world. We are about to open an office in Sydney, Australia and so need a few new support engineers. We realise that engineers in this field are as scarce as hens teeth or the correct lottery numbers these days, but we thought we would at least post a notice on this list server so that none of our friends and clients around the world can say, "But we wanted that job!" afterwards.
As with any technical support job the pay is never enough, the hours are long and the abuse is standard, but you get to work with some real characters and see laboratories all around the world.
If you do still have a slight interest in joining Anaspec, you can visit our web site for more details on our company and then send us an email.
Thanks for your time and hope to see some of you at a conference or laboratory this year. Cheers
Luc Harmsen ANASPEC South Africa www.anaspec.co.za Tel: +27 11 794 8340 Fax: +27 11 794 8349 P.O. Box 2561 Honeydew 2040 Gauteng, South Africa
We have kinda side stepped the issue of traceability by ensuring the original data is available. All images are collected into the Analysis database by Soft Imaging System. There are others out there, this one happened to be floating around the lab at the time I arrived and seems to work well. The database is automatically split into CD size chunks which serve as our archived originals. Users are given read access via a web browser interface. They can snatch their images when they want, and yes, they can do what they want with them, but they must always know that an original exists under my control if there is ever a question.
As an aside for your storage solution, after CD archiving I move the data to a Snap server (or NAS) and lock the database. Essentially it is a series of Raid 5 hard drives and an ethernet card. I can keep about 5 years on at all times (240G) for under $2k and it is completely scalable. Took 5 min to set up and another 5 to set proper access permissions. It was far cheaper than the jukeboxes we looked into and definitely faster serving the images. It works withing most environments PC Mac, unix.... Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Peter Ingram (by way ofMicroscopyListServer) {p.ingram-at-cellbio.duke.edu} 01/10/03 12:51AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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} Steve:
There was symposium on this very topic at the MSA meeting in Quebec City this past summer. I taped most of it and when I find the time (ha!) the plan is to publish a synopsis in Microscopy Today and on the web.
The bottom line if I recall correctly, is that current Copyright Law currently provides most - but not all - protection....at least in the US. Probably there will be variations country-to-country though.
Peter Ingram Public Policy Chair and Legal Liaison Officer, MSA
} } } } } } } } } } } } } } } } } } This email is confidential and intended solely for the use of the Person(s) } ('the intended recipient') to whom it was addressed. Any views or opinions } presented are solely those of the author and do not necessarily represent } those of the Paterson Institute For Cancer Research or the Christie Hospital } NHS Trust. It may contain information that is privileged & confidential } within the meaning of applicable law. Accordingly any dissemination, } distribution, copying, or other use of this message, or any of its contents, } by any person other than the intended recipient may constitute a breach of } civil or criminal law and is strictly prohibited. If you are NOT the } intended recipient please contact the sender and dispose of this e-mail as } soon as possible.
-- Peter Ingram Sr. Physicist Adj. Professor of Pathology, Duke University Medical Center Box 90319 LaSalle Street Extension DURHAM NC USA 27708-0319
I might be missing something here and wondered if some of you could answer a question for me. If we were to add EDS to our ESEM, we could minimize scattering effects by using helium as the charge suppression gas. It small size and low z won't produce spurious x-rays, has minimal beam spreading, and is ionizable for charge suppression. More importantly, many of you swear by it. Now for the question. How do you keep it out of the detector?? Aren't they sealed, and won't helium pass through the window?? Is it going to trash the detector vacuum?? or is my understanding of defector design flawed?? Maybe the pumps recover the vacuum after while as it diffused back out?? Manufacturers encouraged to chime in here. Thanks
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
Dear all, We have two PhD positions available at the moment. My (rough) translation of the original advert is below, followed by the original in German, for those who prefer that.
are available in the Structural Research Group of the Department of Materials and Geological Sciences, as part of the Special Research Area 595 Fatigue of Functional Ceramics. Employment is limited to 4 years.
Applicants should have a Bachelors or Masters Degree in Materials Science, Physics, or Chemistry, and a desire to work towards a PhD in the interdisciplinary field of piezoelectric ceramics. The primary methods used in the work shall be X-ray diffraction, Transmission electron microscopy and synchrotron radiation (at HASYLAB, Hamburg). There will be opportunities to spend time overseas at partner Institutions.
The Technical University of Darmstadt seeks to increase the proportion of women in the staff, particularly in technical subjects and suitably qualified female candidates are therefore especially encouraged to apply.
Suitably qualified disabled persons will be given preferential treatment.
Payment will be calculated using the BAT scale (pay scale for public sector workers in Germany).
Applications should be sent together with Curriculum Vitae to: Das Präsidium der TU Darmstadt, Karolinenplatz 5, 64289 Darmstadt
All applications should be received before 10th February 2003
Stellenausschreibung
Im Fachbereich Material- und Geowissenschaften, Fachgebiet Strukturforschung sind im Rahmen des Sonderforschungsbereichs 595 Ermüdung von Funktionskeramiken zwei Stellen für
Wissenschaftliche Mitarbeiter(innen) - halbtags
(Kenn-Nr. 1) in einem befristeten Arbeitsverhältnis (4 Jahre) zu besetzen.
Gesucht werden Materialwissenschaftler, Physiker oder Chemiker mit abgeschlossenem Diplom, die eine Dissertation in dem praxisnahen, interdisziplinären Arbeitsgebiet piezoelektrischer Keramiken mitarbeiten möchten. Eingesetzte Methoden sind Röntgenbeugung, Transmissionselektronenmikroskopie und Synchrotronstrahlung (am HASYLAB, Hamburg). Gelegenheit zu längeren Auslandsaufenthalten wird gegeben.
Die Technische Universität Darmstadt strebt eine Erhöhung des Anteils der Frauen am Personal, insbesondere in den technischen Bereichen an und fordert deshalb qualifizierte Frauen nachdrücklich auf, sich zu bewerben.
Schwerbehinderte werden bei gleicher Eignung bevorzugt.
Die Vergütung erfolgt nach dem BAT.
Bewerbungen sind mit den üblichen Unterlagen an das Präsidium der TU Darmstadt, Karolinenplatz 5, 64289 Darmstadt, zu senden.
Bewerbungsfrist: 10. Februar 2002
-- Ian MacLaren Strukturforschung, Materialwissenschaft, TU-Darmstadt, Germany ian.maclaren-at-physics.org / http://homepages.tu-darmstadt.de/~maclaren/
} On Thursday, January 9, 2003, at 05:10 AM, Dorota Wadowska wrote: } } } We are looking into buying a new vacuum evaporator. We would it like } } to be equipped in two electrodes (for carbon and metal evaporation), } } rotary/tilt stage and thickness monitor. Our laboratory deals mainly } } with biological/veterinary samples. I would like to hear your } } recommendations regarding the purchase: equipment servicing and make } } (which one is good which is not). Is Denton the best available on the } } market? Do they still supply DV 502A? } } Dear Dorota, } I have also been investigating this, and I've gotten info about } Cressington, Denton, and Ladd evaporators. The final decision of what } to buy depends on what you want to do. For routine carbon and metal } evaporation, Cressington makes a relatively inexpensive (~$4k) benchtop } carbon coater, which uses only a rotary pump (sold separately) and } works at a vacuum of ~10^-2 torr. Their high-end turbopumped system } goes for ~$30k. Denton makes a benchtop turbo system that will handle } both carbon and metals for about $20k, and their high-end system for } high-resolution coating--evaporation or sputtering--goes for ~$70k. } Ladd makes a free-standing system with a diffusion pump that will also } handle both carbon and metals for about $20k. Other manufacturers: } Please feel free to contact me off-list. Your requirements for } uniformity of coating and preference for benchtop vs free-standing } models--as well as your budget--will determine your choice.
Yours, Bill Tivol 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
Our service lab does TEM processing and examination of nearly all kinds of biological samples. We have particular expertise with human muscle, nerve, and kidney biopsies, and are a CAP certified facility. We also do a lot of work with Immunobed and JB-4 resins. We are happy to work with other universities and private industry. For specific information about pricing, turn-around time, etc., please contact our lab administrator, Donna Craft.
Ralph Common Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824
ralph.common-at-ht.msu.edu
Donna Craft Michigan State University Division of Human Pathology A608 East Fee Hall East Lansing, MI 48824
craftd-at-msu.edu
517-355-7558 -------------
On Thu, 9 Jan 2003, William McManus wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear group: } } The EM Facility at Utah State University is being closed, March 1st 2003 } due to budget cuts to the University. I am looking for other facilities } which will do service work, and pricing rates for that work. Some of } the work will be for University personnel, but most will be for off-site } industrial users. Please let me know if you can do work for one group } or the other or both. This includes both biological and materials; TEM } and SEM (high res, and environmental) and edx. } } Thank you for any help in this matter. } } Bill } } William McManus } Supervisor } Microscopy Facility } Utah State University } Logan UT 84322-5305 } } billemac-at-biology.usu.edu } office 435-797-1920 } cell 435-757-2976 } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Scott's got the right idea as far as I'm concerned. Whenever I finish a report I provide a CDROM with the report and [digital copies of] all the original images obtained during the project. I also include in the names of the edited images the serial numbers of the originals from which they were prepared. And I make only globally applied changes to contrast, brightness, tone, etc. in addition to rotation and cropping or enlargement. That means the originals (including the Mavica's thumbnails and HTML index) are available for all to compare with my versions. And I place another copy of each CDROM with the file in case the computer should go belly up.
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
Okay folks - you know the drill. A client walks into a materials lab with a biological problem or vice versa. I work in a materials lab with SEM and LM and had a client walk in today with a biological question. So I am out of my element. Can you help?
It seems this man's daughter is suffering from some kind of skin ailment and she has dug stuff out of MANY of the sores on her skin. The doctors in her HMO have not offered any relief. Her father (from another department on this campus) is quite determined to find out what the problem might be. He would take no for an answer and would accept whatever help we could give him.
I examined the samples he brought in have posted images on our web server at ftp://www.marl.iastate.edu/What_is_it/. Many of them seem to be either bundles or wads of fibers or else little nodules of "stuff" with the fibers running throughout them. I suppose the stuff was secreted by the body, but can't say for sure.
LM pictures show the fibers to be both clear and darkly colored. BSE images and EDS from the SEM show the fibers to be organic in nature. (The nodules contain notable amounts of Na, Cl, and K along with some Si. But the fibers are primarily C and O.) The images don't appear to be like any natural or manmade fibers that I have ever encountered before. I can't find a match in our copy of the Particle Atlas.
Have any of you in all your experience come across such things? I would appreciate whatever expertise you could lend.
Thanking you in advance, Warren
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
A client has approached us asking for assistance in examining some "mercury-wetted" relay contacts. They have had problems with the contacts sticking and would like us to examine a few sets of contacts to see if we can identify the source of the problem.
My question is whether I should be concerned with inserting such samples in the microscope?
I don't intend to parts to be obviously wetted with mercury. However, I presume whatever Hg goes in will probably get volatilized and run out through our diffusion and rotary pump. We have mist traps on the outlet of our rotary pump. Do I need to be concerned about it reacting in bad ways with either the pump oil or components in the scope? (This would be in our conventional SEM rather than our VP-SEM. Its EDS system is down at the moment.)
Thanks in advance for your advice.
Warren
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
The Helium will diffuse through the EDS window and slowly ruin the vacuum in the EDS detector. Fortunately you are running in a vacuum and not applying 1 atm He to the window.
When I was in ED XRF back in Be window days, we would say you could use He instead of vacuum to avoid the absorption of low energy x-rays ( {3 KeV) by air. But those customers who did so eventually had to send their detectors back for repumping. Today's UTWs also leak Helium. You will have slow EDS dewar vacuum degradation if you do this. Another negative is encountered if there is a Ion Pump on the electron gun. Ion pumps do not pump the noble gases well and get flooded.
Ron Vane XEI Scientific
----- Original Message ----- } From: "Scott Whittaker" {Whittaker.scott-at-nmnh.si.edu} To: {microscopy-at-microscopy.com} Sent: Friday, January 10, 2003 6:32 AM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (medvitz-at-pitt.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, January 10, 2003 at 13:13:07 ---------------------------------------------------------------------------
Email: medvitz-at-pitt.edu Name: Neil Medvitz
Organization: University of Pittsburgh
Education: Graduate College
Location: Pittsburgh, PA USA
Question: What is the advantage if any to using gold grids instead of Nickel for post-embedding immunochem.
I'd like to jump in here with a comment or two on Cyberlab that we are developing for ERES compliance (Electronic Record/Electron Signature) as part of validation efforts to be in compliance with FDA regulations.
Cyberlab will store any file type as far as I know, please correct me if you know otherwise. The server ("archive") system can be set up to poll the camera workstation computer whenever and however often the user or "management" decide. This allows for deletion of images that are of no use, such as poor exposure, poor focus, vibration without having to go through all the documentation for a totally useless image.
Once the image is taken off the camera workstation computer it is "archived" by the Cyberlab server "immediately" (users cannot set that on our system) and can be retrieved at a later time; the original can be retained or automatically deleted from the workstation after some user defined period of time. Once retrieved from the server to perform analysis, etc., if it is stored by the same file name, it is given a version number. If it has a different name or file extension it is stored by that new name but linkage to the original is set up. How we will ultimately store terabytes of data over the years remains to be seen as the issue of long term retrieval capability as hardware changes have to be addresssed.
I agree with your statement about assumptions of guilt but we have only to look around us in the business world to see that the need exists. This will only make it more difficult to "beat" the system. Since such systems are designed so that users don't have access to the servers or storage, well that is only a small part of the security of the data that we have to set up. We also have to have workstation computer security so that only authorized users can get access and only to their folders etc. so that the servers see the data as coming from that user. Much more has to be done to reach compliance or whatever rules are ultimately decided on.
Damian Neuberger Senior Research Scientist Baxter Healthcare Corp. PO Box 490 Round Lake, IL 6003 damian_neuberger-at-baxter.com
} Steve, } } Our industry is currently implementing systems to deal with regulatory } agency requirements as stated by Mike Bode in his recent reply. There are a } few companies working on database solutions to solve some of the } traceability issues related to electronic raw data and data manipulation. } I am aware of 2 companies that are marketing database applications to } address these issues (Cyberlab / Nugenesis(SP?)), and there are probably } many more companies. Keep in mind that these applications are not } specifically targeted at imaging data, but rather will address most file } types. In short, the raw data is sent to a server, the database then } archives the file to the database (at some user specified interval). To } view the information the files are "restored" to a specified location where } they can be manipulated. These manipulated files are then returned to the } database using version control methodologies. An audit trail is maintained } which allows for complete tracking of raw data access and manipulations. In } our installation the archived files are maintained on servers making raw } data retrieval a simple affair i.e. no CD's or tapes need be changed / } loaded. } } In my opinion this type of archival system would be very cumbersome and } prohibitively expensive for most EM facilities to implement AND maintain. } } One final note: It's a sad state of affairs when the assumption is made } that these systems are required to prevent fraud. Dishonest people will } always find a way to "beat" the system. It was true for wet chemistry } methods and it is true for digital imaging as well. In my mind these } systems and procedures are required to prevent the inadvertent, unknowing } alteration of raw data. Accidentally deleting or overwriting a file, } "misplacing" a file, or possibly confusing a raw data file with one that you } knowingly manipulated during analysis. I am speculating but I would assume } that most of us have experienced some or possibly all of these situations at } one point in our digital careers. Honest mistakes, yes, but troubling } errors that must be avoided. They waste time, effort, money, and can be } very costly in terms of your reputation and the credibility of your work. } } My two cents worth. } } } John Robson } Boehringer Ingelheim Pharmaceuticals, Inc. } PO Box 368 } 900 Ridgebury Rd } Ridgefield, CT 06877 } } Phone (203)798-5640 } Fax (203)798-5698 } } e-mail jrobson-at-RDG.boehringer-ingelheim.com
If they do, they could plug in a flash memory puck and dump up to a gig of data and walk out with it. Does your system prevent this?
Just a thought.
gary g.
At 10:40 PM 1/10/2003, you wrote:
} John, } } I'd like to jump in here with a comment or two on Cyberlab that we are } developing for ERES compliance (Electronic Record/Electron Signature) as } part of validation efforts to be in compliance with FDA regulations. } } Cyberlab will store any file type as far as I know, please correct me if you } know otherwise. The server ("archive") system can be set up to poll the } camera workstation computer whenever and however often the user or } "management" decide. This allows for deletion of images that are of no use, } such as poor exposure, poor focus, vibration without having to go through } all the documentation for a totally useless image. } } Once the image is taken off the camera workstation computer it is "archived" } by the Cyberlab server "immediately" (users cannot set that on our system) } and can be retrieved at a later time; the original can be retained or } automatically deleted from the workstation after some user defined period of } time. Once retrieved from the server to perform analysis, etc., if it is } stored by the same file name, it is given a version number. If it has a } different name or file extension it is stored by that new name but linkage } to the original is set up. How we will ultimately store terabytes of data } over the years remains to be seen as the issue of long term retrieval } capability as hardware changes have to be addresssed. } } I agree with your statement about assumptions of guilt but we have only to } look around us in the business world to see that the need exists. This will } only make it more difficult to "beat" the system. Since such systems are } designed so that users don't have access to the servers or storage, well } that is only a small part of the security of the data that we have to set } up. We also have to have workstation computer security so that only } authorized users can get access and only to their folders etc. so that the } servers see the data as coming from that user. Much more has to be done to } reach compliance or whatever rules are ultimately decided on. } } Damian Neuberger } Senior Research Scientist } Baxter Healthcare Corp. } PO Box 490 } Round Lake, IL 6003 } damian_neuberger-at-baxter.com } } } } } Steve, } } } } Our industry is currently implementing systems to deal with regulatory } } agency requirements as stated by Mike Bode in his recent reply. There are } a } } few companies working on database solutions to solve some of the } } traceability issues related to electronic raw data and data manipulation. } } I am aware of 2 companies that are marketing database applications to } } address these issues (Cyberlab / Nugenesis(SP?)), and there are probably } } many more companies. Keep in mind that these applications are not } } specifically targeted at imaging data, but rather will address most file } } types. In short, the raw data is sent to a server, the database then } } archives the file to the database (at some user specified interval). To } } view the information the files are "restored" to a specified location } where } } they can be manipulated. These manipulated files are then returned to the } } database using version control methodologies. An audit trail is } maintained } } which allows for complete tracking of raw data access and manipulations. } In } } our installation the archived files are maintained on servers making raw } } data retrieval a simple affair i.e. no CD's or tapes need be changed / } } loaded. } } } } In my opinion this type of archival system would be very cumbersome and } } prohibitively expensive for most EM facilities to implement AND maintain. } } } } One final note: It's a sad state of affairs when the assumption is made } } that these systems are required to prevent fraud. Dishonest people will } } always find a way to "beat" the system. It was true for wet chemistry } } methods and it is true for digital imaging as well. In my mind these } } systems and procedures are required to prevent the inadvertent, unknowing } } alteration of raw data. Accidentally deleting or overwriting a file, } } "misplacing" a file, or possibly confusing a raw data file with one that } you } } knowingly manipulated during analysis. I am speculating but I would } assume } } that most of us have experienced some or possibly all of these situations } at } } one point in our digital careers. Honest mistakes, yes, but troubling } } errors that must be avoided. They waste time, effort, money, and can be } } very costly in terms of your reputation and the credibility of your work. } } } } My two cents worth. } } } } } } John Robson } } Boehringer Ingelheim Pharmaceuticals, Inc. } } PO Box 368 } } 900 Ridgebury Rd } } Ridgefield, CT 06877 } } } } Phone (203)798-5640 } } Fax (203)798-5698 } } } } e-mail jrobson-at-RDG.boehringer-ingelheim.com
It should not be necessary to scrape the sample off the glass to process it. There was a discussion on this point recently, but it may not have been on this list - I subscribe to several. You can fix in a primary aldehyde fixative, followed by a buffer rinse and then secondary fixation with osmium tetroxide in an aqueous buffer . After rinsing in water, dehydrate through graded alcohols finishing with several changes of dry absolute alcohol or with propylene oxide, then in a disposable dish pour a layer of Epon (or other epoxy resin) onto the slide. Change the Epon at least once, then use Epon plus accelerator. Cut the bottom off a Beem capsule and place the capsule tube over the area of interest, then polymerise the whole thing. Next day, fill the Beem capsule with accelerated Epon and polymerise. Pulling the Beem capsule off the glass after cooling should bring the sample with it, or it is possible to chip the glass away before sectioning. I am assuming the glass is a slide; if it's not then it should be possible to modify this technique. Use a fume hood and glove;, all the reagents except alcohol are dangerous, especially OsO4. Hope this helps.
Lesley Weston.
on 09/01/2003 7:18 PM, Kim Handley at khan030-at-ec.auckland.ac.nz wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello all, } } I am a graduate student with the University of Auckland requiring advice on } (1) } the post-fixation stage for TEM prep., (2) any advice concerning the treatment } of fragile sample material for TEM (standard viewing or shadowing). } } I am using TEM to look at freshly grown microstromatolitic films from a } geothermal brine where the primary mineral component is colloidal silica, with } a minor component of sulphur. The objective is to study biomineralisation. } We have scrape one set of my samples from their glass substrate (a terribly } destructive stage as far a texture goes as the samples are very brittle and } porous), dehydrated them through an ethanol series and added osmium tetroxide } as a post-fixative (after initial fixation with 2.5% glutaraldehyde/buffer), } however in similar studies the osmium tetroxide stage has been omitted - any } ideas why? } } I would be greatful for any advice. } } Kind Regards, } Kim } -- } Kim M. Handley } Geology Department } University of Auckland } k.handley-at-auckland.ac.nz } } } }
Neil Medvitz asked: ========================================================================= Question: What is the advantage if any to using gold grids instead of Nickel for post-embedding immunochem. =========================================================================== There are several advantages which could be of greater or lesser importance depending on individual situations:
a) When a pair of normal antimagnetic stainless steel tweezers are used to hold a typical nickel grid during the typical immunoreaction, you have two dissimilar metals in contact in a low pH environment and you get a current flow (this is the basis of electrochemistry). And this current flow is thought to stunt the strength of the immuno reaction. I don't know to what degree that is published but this is what customers have told me over the years as to why they have concerns about nickel grids.
b) Gold grids don't suffer from this problems and they are far more inert under these same conditions than nickel. Consequently, there is for some a preference for gold. But gold is softer, and the grids tend to be a bit less self -supporting and therefore more difficult to work with, whereas nickel is more stiff, and does not bend as readily, that being the reason why some workers, at the end of the day, prefer nickel.
One can get around the electrochemistry issues when using Ni grids, however, if instead of using the normal antimagnetic stainless steel tweezer, gold plated tweezers are used. See URL http://www.2spi.com/catalog/tweezers/precision.html The gold plating acts as a passivation layer on the antimagnetic stainless steel tweezers, killing off the chances for an electro chemical reaction, also leading sometimes to corrosion product in one's samples.
Disclaimer: SPI Supplies offers both gold and nickel grids as well a range of tweezers including gold plated tweezers.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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Warren, I've not experienced it myself, but I've heard of the gun being shorted by the Hg vapor generated when the beam hits the Hg and heats it. Your VP system might be a better bet with the increased pressure and some sort of through-put of gas.
Ken Converse owner Quality Images third party SEM service Delta, PA
Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A client has approached us asking for assistance in examining some } "mercury-wetted" relay contacts. They have had problems with the } contacts sticking and would like us to examine a few sets of contacts } to see if we can identify the source of the problem. } } My question is whether I should be concerned with inserting such } samples in the microscope? } } I don't intend to parts to be obviously wetted with mercury. However, } I presume whatever Hg goes in will probably get volatilized and run } out through our diffusion and rotary pump. We have mist traps on the } outlet of our rotary pump. Do I need to be concerned about it reacting } in bad ways with either the pump oil or components in the scope? (This } would be in our conventional SEM rather than our VP-SEM. Its EDS } system is down at the moment.) } } Thanks in advance for your advice. } } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of } materials } Computer applications and networking } } } }
We put an Oxford GEM detector on our VP-SEM back in 1994. We routinely run Helium as the gas when we are running the scope. We switch over to air for idle periods. I suppose the column is filled with helium about 20 hours per week on average. (Sometimes it is operated in high vacuum mode and sometimes it is plain idle.) I don't think we had any changes in LN2 consumption over the years. We refilled the dewar twice a week and could probably count on it going 5 days between fills before going dry. The low LN2 alarm would go off before that happened.
The helium atmosphere certainly does much to improve the quality of the image and it also reduces the scattering effect on the x-ray signal. I can pass on details and images to prove the point. In fact I had to prove the point to one of our service engineers. They hadn't heard that the type of atmosphere made that big a difference.
During the same time we had a Kevex Quantum detector on our conventional SEM. We have 170 liter LN2 dewars in each scope room for filling the EDS dewars and we can count on both of the big dewars to run out within days of each other. I don't know how dependent that is on evaporation from the big dewars and how much of it is due to consumption in the EDS systems.
So I too now wonder what the manufacturers would say. Are some detector systems immune from He diffusion while components in other system render them more susceptible?
Warren
At 07:48 PM 1/10/03 -0800, you wrote:
} Scott, } } The Helium will diffuse through the EDS window and slowly ruin the } vacuum in the EDS detector. Fortunately you are running in a vacuum } and not applying 1 atm He to the window. } } When I was in ED XRF back in Be window days, we would say you could } use He instead of vacuum to avoid the absorption of low energy x-rays } ( {3 KeV) by air. But those customers who did so eventually had to send } their detectors back for repumping. Today's UTWs also leak Helium. } You will have slow EDS dewar vacuum degradation if you do this. } Another negative is encountered if there is a Ion Pump on the electron } gun. Ion pumps do not pump the noble gases well and get flooded. } } Ron Vane } XEI Scientific } } } ----- Original Message ----- } } From: "Scott Whittaker" {Whittaker.scott-at-nmnh.si.edu} } } To: {microscopy-at-microscopy.com} } } Sent: Friday, January 10, 2003 6:32 AM } } } } I might be missing something here and wondered if some of you could } } answer a question for me. If we were to add EDS to our ESEM, we could } } minimize scattering effects by using helium as the charge suppression } } gas. It small size and low z won't produce spurious x-rays, has minimal } } beam spreading, and is ionizable for charge suppression. More } } importantly, many of you swear by it. Now for the question. How do you } } keep it out of the detector?? Aren't they sealed, and won't helium pass } } through the window? Is it going to trash the detector vacuum? or is my } } understanding of detector design flawed? Maybe the pumps recover the } } vacuum after while as it diffused back out? Manufacturers encouraged to } } chime in here. } } Thanks } } } } Scott Whittaker } } Laboratories of Analytical Biology } } Smithsonian Institution } } National Museum of Natural History } } PO Box 37012 MRC104 } } Washington DC 20013-7012 } } 202-357-1651
This is very odd. I looked at the SEM images wondering if the fibres would look fungal. They do not look fungal to me. They look more like paper or cotton and some synthetic fibres (ribbed longitudinally). Do you think we are looking mostly at the material used to collect the samples?
Dave
On Fri, 10 Jan 2003 17:32:00 -0600 Warren E Straszheim {wesaia-at-iastate.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Okay folks - you know the drill. A client walks into a materials lab with a } biological problem or vice versa. I work in a materials lab with SEM and LM } and had a client walk in today with a biological question. So I am out of } my element. Can you help? } } It seems this man's daughter is suffering from some kind of skin ailment } and she has dug stuff out of MANY of the sores on her skin. The doctors in } her HMO have not offered any relief. Her father (from another department on } this campus) is quite determined to find out what the problem might be. He } would take no for an answer and would accept whatever help we could give him. } } I examined the samples he brought in have posted images on our web server } at ftp://www.marl.iastate.edu/What_is_it/. Many of them seem to be either } bundles or wads of fibers or else little nodules of "stuff" with the fibers } running throughout them. I suppose the stuff was secreted by the body, but } can't say for sure. } } LM pictures show the fibers to be both clear and darkly colored. BSE images } and EDS from the SEM show the fibers to be organic in nature. (The nodules } contain notable amounts of Na, Cl, and K along with some Si. But the fibers } are primarily C and O.) The images don't appear to be like any natural or } manmade fibers that I have ever encountered before. I can't find a match in } our copy of the Particle Atlas. } } Have any of you in all your experience come across such things? I would } appreciate whatever expertise you could lend. } } Thanking you in advance, } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Gold grids are a good choice for on-grid immunodetection when using conventional immunogold reagents (with a size suited for visualization without enhancement, for instance 10 nm gold particles). Gold grids are virtually chemically inert and thus won't easily interfere with components in incubation solutions. There is one exception: silver enhancement. Since gold particles act as catalysts in the deposition of metallic silver, gold grids may become covered with metallic silver as well, locally exhausting the enhancement reagents and leading to non-reproducible results. Nickel, as far as we have tested, does not seem to act as a catalyst for silver enhancement and in a practical sense is also chemically inert towards incubation solutions. In our experience handling such grids with non-magnetic tweezers (or better, with platinum loops) has never been a reason for doubting the immuno results. The main downside to using nickel grids is that they influence the electron beam and cause astigmatism making more frequent adjustments necessary while observing specimens in the TEM.
Electrochemical phenomena may already occur whenever a metal surface is brought into contact with a solution. It does not necessarily involve a second (different) metal. Whether this actually results in a chemical reaction depends a.o. on the redox potential of the components involved and whether the overall conditions are favorable for a reaction to occur.
I would be interested to learn if there are any listers who would be willing to comment on the issue of electrochemical reactions influencing immunodetection. Any documented experience may teach us how to obtain better results.
Jan Leunissen Aurion - ImmunoGold Reagents and Accessories http://www.aurion.nl
Dear Sirs, I would like to know if somebody can help me to find the procedures for the baking of this field emission (JSM6400F). We have the connections to the heaters, we need to know if we have to dismount something in the gun before baking and if there is a difference between baking time on the gun and the column. All other information are well accepted.
I have no experience with the JSM6400F, however that said, I would strongly suspect that the gun bake time should be the longest in the system. The reason is that residual gas will tend to be driven from the hottest to the coolest region of the system, and for any microscope the best vacuum is desired in the gun. For comparison, the bakeout procedure on our SEM (CamScan 44FE) is: entire system, pumped by diffusion pump /w cold trap, all heaters on - 1 day cool lower ion pump; isolate from chamber; pump column+gun+upper ion pump with lower ion pump - 1 day cool upper ion pump; seal upper/lower pumping line; pump with both ion pumps - 1 day finally, cool gun assembly.
hope this helps, Ben Simkin (simkin-at-egr.msu.edu)
I am looking for a manual (operation instructions and technical specs.) for an Edwards E306 thermal evaporator. Of course I will pay for copy-costs and postage. If you can help me with that, please contact me directly: stefan.geimer-at-uni-koeln.de
Morning Warren, While on one of my searches to attempt to insure that our new FEI ESEM remains as uncontaminated as possible, I came across the following related to mercury in amalgams and SEM/vacuum:
http://www.gbg.bonet.se/bwf/art/instability.html
I can't speak to the validity of the experimentation, but I have bookmarked the site.
Hope this helps,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] Sent: Friday, January 10, 2003 6:39 PM To: Microscopy-at-sparc5.microscopy.com
A client has approached us asking for assistance in examining some "mercury-wetted" relay contacts. They have had problems with the contacts sticking and would like us to examine a few sets of contacts to see if we can identify the source of the problem.
My question is whether I should be concerned with inserting such samples in the microscope?
I don't intend to parts to be obviously wetted with mercury. However, I presume whatever Hg goes in will probably get volatilized and run out through our diffusion and rotary pump. We have mist traps on the outlet of our rotary pump. Do I need to be concerned about it reacting in bad ways with either the pump oil or components in the scope? (This would be in our conventional SEM rather than our VP-SEM. Its EDS system is down at the moment.)
Thanks in advance for your advice.
Warren
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Why not just call JEOL and talk to a service engineer? They are very helpful. You do not have to be on a service contract to get their help either.
Ron L
-----Original Message----- } From: Benjamin - Simkin [mailto:simkin-at-egr.msu.edu] Sent: Monday, January 13, 2003 7:02 AM To: Microscopy-at-sparc5.microscopy.com; b.rapillo-at-mclink.it
I have no experience with the JSM6400F, however that said, I would strongly suspect that the gun bake time should be the longest in the system. The reason is that residual gas will tend to be driven from the hottest to the coolest region of the system, and for any microscope the best vacuum is desired in the gun. For comparison, the bakeout procedure on our SEM (CamScan 44FE) is: entire system, pumped by diffusion pump /w cold trap, all heaters on - 1 day cool lower ion pump; isolate from chamber; pump column+gun+upper ion pump with lower ion pump - 1 day cool upper ion pump; seal upper/lower pumping line; pump with both ion pumps - 1 day finally, cool gun assembly.
hope this helps, Ben Simkin (simkin-at-egr.msu.edu)
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (burtonjoyner-at-insightbb.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Monday, January 13, 2003 at 09:22:02 ---------------------------------------------------------------------------
Question: I am new to electron microscopy and have the opportunity to use the TEM at UK. Are there any comprehensive TEM sample preparation references in print? I am not having much luck finding good resources.
We've never had any trouble using Ni for immuno. Cu sometimes can react with reagents, particularly if you incubate overnignt, leaving blobs of copper sulfate (they turn greenish). As for Au, use them for growing cells on support films because the Au is not toxic to the cells. However, I think Au is overkill for immuno; Au grids are EXPENSIVE. If you have some sort of really corrosive treatment (why would you???), you might consider Au.
You should use several grids (at least 2; we use 3) for every different condition (different antibody--positive & negative, different antibody dilution, different tissue/cell type, etc). This means you'll be using lots of grids, which would be expensive if you're using Au ones.
Good luck, Sara Miller
On Fri, 10 Jan 2003, by way of MicroscopyListServer wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (medvitz-at-pitt.edu) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, } January 10, 2003 at 13:13:07 } --------------------------------------------------------------------------- } } Email: medvitz-at-pitt.edu } Name: Neil Medvitz } } Organization: University of Pittsburgh } } Education: Graduate College } } Location: Pittsburgh, PA USA } } Question: What is the advantage if any to using gold grids instead of } Nickel for post-embedding immunochem. } } --------------------------------------------------------------------------- } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
As always, Chuck has some good ideas. One additional one is that we don't use forceps at all. We use loops made with either Ni or Au wire (like the wire you put into the vacuum evaporator) glued onto sticks (we use plastic disposable loops normally used for spreading bacteria onto plates--with the plastic loop cut off). At the very end, we pick up the grid with forceps and dry it.
The loops are made with 1-2 inches of wire, wrapped around a drill bit just larger than the grid and twisted tightly to make a loop at one end. The other end is wrapped around the end of a plastic handle and glued with epoxy.
The advantages are that loops are reusable, and there's less likelyhood of bending the grids. Just make sure you wash grids between solutions sufficiently. We use 5-6 washes (drops of buffer--which is also probably overkill).
Good luck, Sara Miller
On Sat, 11 Jan 2003, Garber, Charles A. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } -- [ From: Garber, Charles A. * EMC.Ver #3.1 ] -- } } Neil Medvitz asked: } ========================================================================= } Question: What is the advantage if any to using gold grids instead of } Nickel for post-embedding immunochem. } =========================================================================== } There are several advantages which could be of greater or lesser importance } depending on individual situations: } } a) When a pair of normal antimagnetic stainless steel tweezers are used to } hold a typical nickel grid during the typical immunoreaction, you have two } dissimilar metals in contact in a low pH environment and you get a current } flow (this is the basis of electrochemistry). And this current flow is } thought to stunt the strength of the immuno reaction. I don't know to what } degree that is published but this is what customers have told me over the } years as to why they have concerns about nickel grids. } } b) Gold grids don't suffer from this problems and they are far more inert } under these same conditions than nickel. Consequently, there is for some a } preference for gold. But gold is softer, and the grids tend to be a bit } less self -supporting and therefore more difficult to work with, whereas } nickel is more stiff, and does not bend as readily, that being the reason } why some workers, at the end of the day, prefer nickel. } } One can get around the electrochemistry issues when using Ni grids, however, } if instead of using the normal antimagnetic stainless steel tweezer, gold } plated tweezers are used. See URL } http://www.2spi.com/catalog/tweezers/precision.html } The gold plating acts as a passivation layer on the antimagnetic stainless } steel tweezers, killing off the chances for an electro chemical reaction, } also leading sometimes to corrosion product in one's samples. } } Disclaimer: SPI Supplies offers both gold and nickel grids as well a range } of tweezers including gold plated tweezers. } } Chuck } } PS: Remember that we are striving to be 100% paperless, therefore there } are no paper copies kept of this correspondence. Please be sure to always } reply by way of "reply" on your software so that the entire string of } correspondence can be kept in one place. } ============================================ } } Charles A. Garber, Ph. D. Ph: 1-610-436-5400 } President 1-800-2424-SPI } SPI SUPPLIES FAX: 1-610-436-5755 } PO BOX 656 e-mail:cgarber-at-2spi.com } West Chester, PA 19381-0656 USA } Cust.Service: spi2spi-at-2spi.com } } Look for us! } ######################## } WWW: http://www.2spi.com } ######################## } ============================================ } } } } }
Sara E. Miller, Ph. D. P. O. Box 3712 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-3265
Have you considered putting the device into a vacuum system first to get any residual Hg off the surfaces prior to EM? That is, a vacuum system that is not the SEM column.
Peter Tomic Anadigics, Inc.
-----Original Message----- } From: qualityimages [mailto:qualityimages-at-netrax.net] Sent: Saturday, January 11, 2003 5:31 PM To: Warren E Straszheim; Microscopy
Warren, I've not experienced it myself, but I've heard of the gun being shorted by the Hg vapor generated when the beam hits the Hg and heats it. Your VP system might be a better bet with the increased pressure and some sort of through-put of gas.
Ken Converse owner Quality Images third party SEM service Delta, PA
Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A client has approached us asking for assistance in examining some } "mercury-wetted" relay contacts. They have had problems with the } contacts sticking and would like us to examine a few sets of contacts } to see if we can identify the source of the problem. } } My question is whether I should be concerned with inserting such } samples in the microscope? } } I don't intend to parts to be obviously wetted with mercury. However, } I presume whatever Hg goes in will probably get volatilized and run } out through our diffusion and rotary pump. We have mist traps on the } outlet of our rotary pump. Do I need to be concerned about it reacting } in bad ways with either the pump oil or components in the scope? (This } would be in our conventional SEM rather than our VP-SEM. Its EDS } system is down at the moment.) } } Thanks in advance for your advice. } } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of } materials } Computer applications and networking } } } }
I am interested in your opinions on the various brands of automatic black and white processors that produce a dry print. For the present, our Center must rely in part on silver emulsion processing as well as digital images. The processors I am looking at are the Colex, the Fujimoto CP31 and CP51. I would consider other brands.
Your comments, if you wish, can be sent to my e-mail address. In advance, I than you for your time.
Dr. Lewis B. Coons, Director Integrated Microscopy Center The University of Memphis Memphis, TN e-mail lcoons-at-memphis.edu
I have read with some interest the recent discussions of methods for cleaning cold cathode discharge gauges. In this connection, it might be of interest to note that there are two different types of CCD gauges. These are described in Sect 3.2.2 of my book Vacuum Methods in Electron Microscopy (for a description see http://www.2spi.com/catalog/books/book48.html). One type, perhaps the first type developed, is the classical 'Penning Gauge'. In modern gauges of this type the anode is usually a loop of tungsten wire that is located in the center of a surrounding cylindrical cathod, as shown in Fig. 3.12. The other type is commonly known as the 'Magneton Gauge". In this gauge the cylindrical case of the gauge is the anode, and the cathode is a rod with circular end plates that is located along the axis of the case, as shown in Fig. 3.13. Magnetron gauges are usually considered to be capable of recording considerably lower pressures than Penning gauges, and to be more stable and more accurate.
It sounds to me as though most of those who commented on the problem were describing procedures for refurbishing a Magnetron gauge, and indeed, my experience also suggests that it is best to replace the central element of this type gauge (the cathode) if it shows significang erosion, and that a local machine shop can produce a new one for you much cheaper than you can purchase one from the gauge manufacturer.
Happy New Year!! -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
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----- Original Message ----- } From: Nanovision S.R.L. {b.rapillo-at-mclink.it} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, January 13, 2003 5:36 AM
Hi Warren
The first care could be to limit evaporation, by use of a cold stage. The Pelltier stage from an ESEM schould be enought, to be just a bit above the melting point of Hg.
The second care could be the use of a cold trap in front of the sample, one like these used in HR-SEM, mounted on the objectif lens, or better, inserting a cold finger in the scope chamber and tilting the sample in it direction. This last configuration would limite the risk to evaporate something in the electron tube. Of coarse, this trap have to be cleaned after the observation, and the mounting of such a cold finger depends of what free port you have on the scope in tilting direction.
Third, use the lowest beam current possible (imaging or EDS ?).
An other care could be to pack the sample in Al foil, with a hole at the place you want to observe, to limit direct relation between the rest of the sample and the SEM vaccum. The vapor flux is proportionnal to the surface of metal exposed to the vaccum.
I don't think that you will have a strong evaporation rate in the vacuum level of a SEM, at room temperature or at -20C. The 10 to 200 pA beam current from a SEM in current observation conditions are far from the that of a MBE gun. Have a look at the vapor pressure curves, or at Holland's book about thin film technologie. Have a look at the vapor pressure curves, or at Holland's books about thin film technology. He gives the way to calculate the vapor flux.
J. Faerber IPCMS-GSI (Institut de Physique et Chimie des Matériaux de Strasbourg Groupe Surface et Interfaces) 23, rue de Loess ; BP43 67034 Strasbourg CEDEX 2 France
} } } -----Original Message----- } } From: Warren E Straszheim [mailto:wesaia-at-iastate.edu] } Sent: Friday, January 10, 2003 6:39 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM: Examining mercury-wetted contacts } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } A client has approached us asking for assistance in examining some } "mercury-wetted" relay contacts. They have had problems with the contacts } sticking and would like us to examine a few sets of contacts to see if we } can identify the source of the problem. } } My question is whether I should be concerned with inserting such samples in } the microscope? } } I don't intend to parts to be obviously wetted with mercury. However, I } presume whatever Hg goes in will probably get volatilized and run out } through our diffusion and rotary pump. We have mist traps on the outlet of } our rotary pump. Do I need to be concerned about it reacting in bad ways } with either the pump oil or components in the scope? (This would be in our } conventional SEM rather than our VP-SEM. Its EDS system is down at the } moment.) } } Thanks in advance for your advice. } } Warren } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 23 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of } materials } Computer applications and networking } } }
You'd better restrict yourself to techniques that are specific to the materials you need to prepare for TEM. Talk to someone knowledgeable at UK. Tell the Microscopy list the kind of materials you need to work on.
At 9:26 AM -0600 1/13/03, by way of MicroscopyListServer wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ----------------------------------------- Russell E. Cook, Ph.D. Electron Microscopy Center Argonne National Laboratory Materials Science Division, Building 212 9700 South Cass Avenue Argonne, IL 60439-4838 USA Telephone: 630-252-7194 FAX: 630-252-4289 recook-at-anl.gov
I have always just used scissors to cut mica to the desired shape and size.
Roger Moretz, PhD Dept of Toxicology BI Pharmaceuticals -- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I am familiar with the techniques of cleaving mica, but what is the } recommended method for cutting mica to a desired shape or size? } } Linda Rinko } Design Engineer } Harvard Medical School } 060 Seeley G. Mudd } Boston, MA 02115 } 617-432-2052 } } }
there is one more possibility - gold-gilded copper grids by Gilder. I buy mine from Electron Microscopy Sciences and like them for most of post-embedding labeling, except silver-enhancement of course. They have proved sturdy and easy to handle and do not seem to react with PBS. They are not much more expensive than Ni grids. I dislike Ni grids for the reasons Jan and others listed, but pure gold ones were too flimsy.
Vlad
} Email: medvitz-at-pitt.edu } Name: Neil Medvitz } } Organization: University of Pittsburgh } } Education: Graduate College } } Location: Pittsburgh, PA USA } } Question: What is the advantage if any to using gold grids instead } of Nickel for post-embedding immunochem. } } ---------------------------------------------------------------------------
-- -------------------------------------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
We recently obtained an older metallograph, namely an Aus Jena Neophot 21, originally sold by Leco Corp. One of the lamps uses a halogen bulb with an extended base. We are having trouble finding a supplier for this type of bulb. If there are any Neophot 21 users out there that have found a bulb supplier, a recommendation would be greatly appreciated. Thanks in advance.
Joseph
Joseph M. Oparowski Center for Materials Science - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone:(508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
Jibao He wrote: ============================================================================ ==== A my friend wants to know which company can produce High Oriented Pyrolyzed Graphite slides used for AFM? or where he can order it? ============================================================================ ==== SPI Supplies has offered highly ordered pyrolytic graphite (HOPG) for many years for AFM applications, the details for which can be found on URL http://www.2spi.com/catalog/new/hopgsub.shtml
A number of custom sizes are shown, but we can make just about any specific size one might want. It is not easy to cut down to size without the special equipment and experience, so it is better to order to the size you want instead of a larger size you might try to cut down.
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 ######################## ============================================
Linda Rinko wrote: ============================================================================ == I am familiar with the techniques of cleaving mica, but what is the recommended method for cutting mica to a desired shape or size? ============================================================================ == One must first understand that mica is very very hard and cutting a thin sheet of mica is nearly like cutting a strip of equal thickness of steel! So no matter what the edge is that you would be using for the cutting, it is going to lose its sharpness very quickly, but just how quickly will depend on the thickness of the mica being cut.
All of the mica pieces described on the SPI Supplies website See URL http://www.2spi.com/catalog/submat/mic_shet.shtml are prepared by die cutting, other wise there will be delamination at the edges and also, a large generation of particulates. Carbide tools are always used for the die cutting, and that is why there is sometimes a not-so -small tooling charge when special sizes or shapes are needed. This is also why the mica sheets and discs sold by SPI Supplies have edges far more free of fracturing and splitting vs. other methods of cutting.
We are often times asked how mica purchased from SPI Supplies can be cut down to smaller pieces. And the answer depends on the thickness of the piece being cut.
Thickness is less than 15 mils (0.015" or 0.381 mm): For sheets of this thickness or less, one can use either a very sharp scissors or an Exacto type knife or even a single edge razor blade. Be sure to wear good eye protection (which should also be worn by any one nearby watching). One can expect some edge fracturing and splitting , the amount depending on the sharpness of the scissors or blade. The splitting can be minimized by holding down the mica, as it is being cut with a ruler or other "straight edge" instrument, the higher the pressure applied , the better the result (e.g. less fracturing). The thinnest mica cleavings can be scissors cut almost as if they were a piece of paper.
Thickness is greater than 15 mils (0.015" or 0.381 mm) but less than 30 mils (0.030" or 0.762 mm): Scissors and razor blades will not work. The only possibility is to use a guillotine type cutter perhaps even a paper cutter. There will be a strong tendency for the mica to edge fracture, but that can be minimized by applying higher downward pressures to the side of the mica on the flat cutting surface. It can be further minimized by being sure that the blade is the sharpest possible.
Thickness is greater than 30 mils (0.030" or 0.762 mm): We do not believe that anyone can do a credible job cutting such mica pieces with out resorting to the use of good tooling and die cutting.
Just remember that any cutting edge that you might use to cut the mica is going to be become dull quickly. While razor blades and even scissors can be discarded and new ones then used, one normally does not think of a paper cutter as being a disposable instrument. At the same time, we are not aware of any service that resharpens the guillotine blades from worn out paper cutters.
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 ######################## ============================================
Try Kreonite and Ilford processors. Used ones can be found at:
http://dunningphoto.com/used.html
http://www.kreonite.com/procpm2.htm
Kreonite has been around a long time.
gary g.
At 09:32 AM 1/23/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Greetings, } } We recently obtained an older metallograph, namely an Aus Jena Neophot 21, } originally sold by Leco Corp. One of the lamps uses a halogen bulb with an } extended base. We are having trouble finding a supplier for this type of } bulb. If there are any Neophot 21 users out there that have found a bulb } supplier, a recommendation would be greatly appreciated. Thanks in advance. } } } Joseph } } Joseph M. Oparowski } Center for Materials Science - Consulting and Failure Analysis } Bose Corporation Joseph_Oparowski-at-bose.com } The Mountain, M/S 415 Phone:(508) 766-1371 } Framingham, MA 01701-9168 Fax: (508) 766-1313
Also check out Mohrpro processors. They are available in various sizes. Reliable and easy to use. http://www.mohrpro.com/
George
George Laing National Graphic Supply ph 800.223.7130 x3109 f 800.832.2205 email scisales-at-ngscorp.com
Dear Listers:
I am interested in your opinions on the various brands of automatic black and white processors that produce a dry print. For the present, our Center must rely in part on silver emulsion processing as well as digital images. The processors I am looking at are the Colex, the Fujimoto CP31 and CP51. I would consider other brands.
Your comments, if you wish, can be sent to my e-mail address. In advance, I than you for your time.
Dr. Lewis B. Coons, Director Integrated Microscopy Center The University of Memphis Memphis, TN e-mail lcoons-at-memphis.edu
I don't know if they supply the bulb you are looking for, but we buy most of our bulbs from Gray Supply. You can find them at 1-800-TOP-BULB (1-800-867-2852) or on the web at www.topbulb.com . They have a large supply of specialty bulbs and one of them may fit your application.
Mike
=========================================== Michael D. Standing Electron Microscopy Technologist Brigham Young University 401 WIDB Provo, UT 84602
-----Original Message----- } From: Oparowski, Joseph [mailto:Joseph_Oparowski-at-bose.com] Sent: Tuesday, January 14, 2003 11:44 AM To: 'microscopy-at-sparc5.microscopy.com' Cc: Shaw, Douglas
Greetings,
We recently obtained an older metallograph, namely an Aus Jena Neophot 21, originally sold by Leco Corp. One of the lamps uses a halogen bulb with an extended base. We are having trouble finding a supplier for this type of bulb. If there are any Neophot 21 users out there that have found a bulb supplier, a recommendation would be greatly appreciated. Thanks in advance.
Joseph
Joseph M. Oparowski Center for Materials Science - Consulting and Failure Analysis Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone:(508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
On Tuesday, January 14, 2003, at 06:58 AM, Linda Rinko wrote:
} I am familiar with the techniques of cleaving mica, but what is the } recommended method for cutting mica to a desired shape or size? } Dear Linda, I buy mica in ~1 cm x 3 cm x 1 mm pieces, then just use a scissors to cut a piece lengthwise. I also cut off a corner so I can tell which side has been carbon coated. Yours, Bill Tivol 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
For more than ten years we've been using a Hammamatsu SIT camera for imaging at video rates at low light fluorescence. However,the camera has serious vignetting and noise. Also, the tube has gotten dirty (dust) and we can't clean much of it.
We'd like to replace this camera with one with a flatter field, similar sensitivity, and at least 15 FPS at 640X480 or greater.
Yes, the Roper/Cooke/Hammamatsu $16k cooled CCDs could do this in many of the situation we image, but what about for much less money?
Thanks!
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
I am doing work on fluorescent staining of blood vessel tumor cross-sections. I was told that blood vessels will autofluoresce without the aid of staining. Is this true and if so at what wavelength will it fluoresce? Currently we are double staining to get our results (staining vessel cells and proteins inside of the cells), but if we only have to stain once, it would be beneficial. If the autofluorescing does not work well enough, does anyone have any ideas on what colors/stains would work for the double staining of cells that makeup the blood vessels (endothelial) and then tagging something within those cells? Right now we are using texas red and I am getting a ton of background noise (red) that makes it to difficult to see. Any information can be mailed to me directly, please, at bened008-at-umn.edu
Thanks,
Ryan Benedict Oral Sciences Department University of Minnesota 612-626-3562
I had to face the same problem a while ago (got an old Neophot 21). What I figured out is that the prices up here in Canada is much lower, compared to the States. The company I bought the bulbs from is Microlites Scientific. You can call them at: 1-800-263-8902 or 416-2995301. Speaking about the scope, does anybody have one for parts or has some spare parts for the machine?
Regards,
Vlad Igoshev, Ph.D.
Toronto, Canada
----- Original Message ----- } From: "Oparowski, Joseph" {Joseph_Oparowski-at-bose.com} To: {microscopy-at-sparc5.microscopy.com} Cc: "Shaw, Douglas" {Douglas_Shaw-at-bose.com} Sent: Tuesday, January 14, 2003 1:44 PM
Wil-
May be you could explain to me what mean "Inverted Magnetron" (or point some page in your book)? I got AIM-x (Active Inverted Magnetron) Gauge from Edwards a few years ago. Edwards claimed that this gauge is good up to 10-9 mbar. In recent discussion, somebody (I am sorry, I don't remember the name) claimed that "cold cathode gauge" could not perform well over 10-5 torr. How it may happening that my "inverted magnetron" gauge performed perfectly up to 10-8 torr? Thanks, Sergey.
At 12:45 PM 1/13/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Michael, you are unlikely to find a camera sensitive enough to decently record fluorescence image at 15 frames per second, with software, for much less than $16K. However if, you agree to wait for 12 long seconds while image downloads from the camera, then check out http://www.starlight-xpress.co.uk/ . They have Peltier cooled 1300 x 1030 CCD camera with SONY ICX085AL sensor, at $2,500. Ask for industrial version, it is designed for long duty cycle. That's the least expensive one of such resolution on the market. And if you use it in 650 x 515 mode, (which you are looking for), the download takes only 3 seconds. Another company, SBIG, offers the least expensive 640 x 480 camera with Texas Instrument TC37 sensor, also Peltier cooled, and also very sensitive, with integrated color filter wheel, at $1,700. You can choose from a huge variety of filters. Check www.sbig.com . Both cameras come as a complete package with the power supply, camera/filters control and image processing software, and all cables. SBIG software is somewhat better for processing. Control capabilities are similar.
Disclaimer: I have no commercial interest in neither Starlight Express nor SBIG companies, other than being a satisfied customer.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: Michael Cammer {cammer-at-aecom.yu.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, January 15, 2003 4:31 PM
Dear All,
We are disposing of a trusty old friend, a Cambridge S250 MkIII. I am keeping several vacuum components and the BSE detector but the remainder could form useful spares for someone. Removal/shipping costs only.
Stu ---------------------- SL Kearns, Earth Sciences University of Bristol Bristol, UK +44 (0)117 954 5421 Stuart.Kearns-at-bristol.ac.uk
We are shortly saying goodbye to two Philips 301 transmission electron microscopes. One has a double tilt facility - the other one is not so high-powered, but might be considered as a source of parts for the first.
Would anyone be interested? If so, please let me know and I will take the matter higher up as regards arrangements.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +-----------------------------------------+ Phone: {direct line +44 (0) 118 9318572 {University internal extension 7867 Fax: +44 (0) 118 9750203 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +-----------------------------------------+
We, too, have the same metallograph (AusJena Neophot 21 from LECO) in our lab. If the other leads listed on the Microscopy Listserver don't pan out, you may want to contact the contractors that service our equipment:
Hi-Point Optical Calibration 567 North Park Road Bellefontaine, Ohio 43311 (937) 592-2641
Bob & Mark Pickering (specialists) Julia Pickering (office clerk)
They purport to be very familiar and experienced with this particular microscope. You may want to keep them in mind for any future spare parts needed for your AusJena metallograph.
Regards,
Stu Smalinskas Senior Metallurgist SKF USA Plymouth, Michigan
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Hi Ryan, Indeed, blood vessels will be autofluorescent and the problem is that they will autofluoresce in all the channels we are usually using e.g. red, green and blue. I am facing a similar problem right now and to solve it you would have to go with a fluorophore that will emit at a wavelenght close to the infra red end of the spectrum. That would mean around 670-700 nm or higher. You would have to avoid the 450-615 part of the spectrum were a lot of biological material will naturally fluoresce. The problem now is to find the (secondary or tertiary) antibody coupled with the probe you have selected (ex: Cy5, Cy 5.5 or others ...coupled Ab) and the other problem is that you can't see the labeling with your eyes because we can't see the infra-red or close wavelenght. So you need a digital camera linked to a computer to see anything. Obviously you also need the right filter on your microscope... Hope this helps. Emmanuelle
I have inherited a EMITECH freeze drier K775X which has never been setup. The instructions are not very good for someone who has never used any kind of freeze drier. It would be helpful to have step by step instructions for this instrument. Actually it looks pretty simple, but that is relative, isn't it?
Can someone help me? You may respond back to me directly.
Thanks.
Winnie
Edwina W. Westbrook Electron Microscopy Laboratories Agriculture Research Station P.O.Box 9061 Virginia State University Petersburg, VA 23806 (804)-524-5659
I created a Power point presentation last November 2002 consisting of several fluorescence micrographs. The file which was rather large ( 90 Mb) was left in my laptop all this time. When I opened it again this week I find that the images are now too dark and I needed to increase brightness by 3- 4 clicks on the brightness icon of Power point.I increased brightness of all the micrographs and copied the file to a CD hoping that the image will not deteriorate there and then compare it with the one in my laptop several weeks from now. Has this happened to anyone else? Is there something I should have done to prevent this?
Any suggestions or comments will be greatly appreciated.
Let me assure you that the images on your laptop have not degraded. If they had, you would have probably encountered an error reading the file, or you might have found a bunch of junk pixels in your images.
The culprit here is probably the brightness setting on your laptop display. Others who do this more than me will quickly tell you that it can be a real bugaboo trying to get monitors set right so that the image displays correctly on them and also prints out correctly on the various printers. I understand that some folks command a pretty good fee for calibrating monitors and printers for print houses.
I would suggest getting a test image to use for setting your monitor. I have a grid of gray swatches ranging from black to white. I set up the controls so black is black, white is white, and I have a pretty good range of mid-tones.
There is also a control panel from Adobe to help generate an ICM file to account for monitor characteristics. I have used it some, but am not real confident about its use.
I have made a test file (testgrad.gif) and the Adobe control panel available on our server. You may reach them by pointing your browser at ftp://www.marl.iastate.edu/ and then navigating to the "Test images" folder. You will need to drop the control applet into your Windows\System folder and then open up the Control Panel. I use it under Windows 98. It should work under other versions, but I cannot vouch for it. You will be on your own.
Hopefully, these will help you consistently setup you laptop so that the appearance stops changing from month to month.
Warren
At 11:11 AM 1/16/03 -0600, you wrote:
} I created a Power point presentation last November 2002 consisting of } several fluorescence micrographs. The file which was rather large ( 90 } Mb) was left in my laptop all this time. When I opened it again this week } I find that the images are now too dark and I needed to increase } brightness by 3- 4 clicks on the brightness icon of Power point.I } increased brightness of all the micrographs and copied the file to a CD } hoping that the image will not deteriorate there and then compare it with } the one in my laptop several weeks from now. Has this happened to anyone } else? Is there something I should have done to prevent this? } } Any suggestions or comments will be greatly appreciated. } } Cora Bucana
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Hello Everyone, I came across a new problem with our Electroscan E3 ESEM, that I've not been able to track down. At times, the image will become very noisy and streaky with really bright streaks across the image making it near impossible to focus. The problem goes away with time by venting and repumping the chamber, or fiddling with the detector ESD hook wire - but I'm not sure exactly what is causing it. Anyone ever seen this before? Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Following the comments I made a few days ago about magnetron cold cathod discharge gauges, several people have asked about 'inverted magnetron gauges'.
As it turns out, in addition to the two most common types of cold cathode discharge gauges that I described in my book, there are at least three others: (1) inverted magnerton gauges; (2) Redhead magnetron gauges; and (3) alphatron CCD gauges. For those interested, these are described in reasonable detail in Sect. 6.7.3 of the book 'Vacuum Technology' (2nd. Ed.) by Alexander Roth (ISBN 0-444-86027-4 (Elsevier))
Briefly, the inverted magnetron has a central anode rod that is surrounded by a cylindrical cathode, as in the ordinary magnetron gauge, but in addition there is an auxiliary cylindrical cathode that surrounds theis 'inner cathode', which modifieds the field distribution at the ends of the anode. These gauges are said to be useful over the range from 10-4 to 10-13 Torr.
The Redhead magnetron gauge is a modification of the inverted magnetron which has perforations in the cathode cylinder to improve conductance. It reportedly is useful down to 10-10 Torr.
The alphatron gauge contains a radioactive source of alpha particles that produce ionazition of gas molecules at high pressures, allowing measurements in the range from 10-3 up to 40 Torr.
May all your vacuum leaks be small ones,
Wil Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
The only explanation for what you are observing is that the screen of your laptop has deteriorated or changed. It is statistically impossible that an image "darkens" while stored on the computer. An image on the computer consists basically of millions of numbers that describe the intensity at each point. It is of course possible, that a number gets corrupted and you get a different brightness at a pixel, but that all pixels deteriorate in the same fashion is statistically impossible. Another possibility could be, that your settings for the brightness in Powerpoint changed.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Corazon D. Bucana [mailto:bucana-at-audumla.mdacc.tmc.edu] Sent: Thursday, January 16, 2003 10:11 AM To: Microscopy-at-sparc5.microscopy.com
I created a Power point presentation last November 2002 consisting of several fluorescence micrographs. The file which was rather large ( 90 Mb) was left in my laptop all this time. When I opened it again this week I find that the images are now too dark and I needed to increase brightness by 3- 4 clicks on the brightness icon of Power point.I increased brightness of all the micrographs and copied the file to a CD hoping that the image will not deteriorate there and then compare it with the one in my laptop several weeks from now. Has this happened to anyone else? Is there something I should have done to prevent this?
Any suggestions or comments will be greatly appreciated.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gtg187h-at-mail.gatech.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, January 16, 2003 at 10:28:03 ---------------------------------------------------------------------------
Email: gtg187h-at-mail.gatech.edu Name: Leslie Smith
Organization: Georgia Instituite of Technology
Education: Undergraduate College
Location: Atlanta, GA, USA
Question: My BioMedical Engineeering class is working ona problem dealing with Electonic Muscle Stimulation. we were wonderng what types of microscopy would be best for viewing muscles. Thanks so much Leslie Smith
Did your gamma change? i.e., brightness and/or contrast? That will make a huge difference. It is totally unlikely that the native files are changed or perturbed. your viewing situation is more likely the problem. Try using the Adobe gamma adjust app to set your display's gamma. Also note that LCDs are not all that great for pictures. Active matrix LCDs are better. CRTs are best...IMO.
gary g.
At 09:11 AM 1/16/2003, you wrote:
} I created a Power point presentation last November 2002 consisting of } several fluorescence micrographs. The file which was rather large ( 90 } Mb) was left in my laptop all this time. When I opened it again this week } I find that the images are now too dark and I needed to increase } brightness by 3- 4 clicks on the brightness icon of Power point.I } increased brightness of all the micrographs and copied the file to a CD } hoping that the image will not deteriorate there and then compare it with } the one in my laptop several weeks from now. Has this happened to anyone } else? Is there something I should have done to prevent this? } } Any suggestions or comments will be greatly appreciated. } } Cora Bucana }
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I created a Power point presentation last November 2002 consisting of } several fluorescence micrographs. The file which was rather large ( 90 Mb) } was left in my laptop all this time. When I opened it again this week I } find that the images are now too dark and I needed to increase brightness } by 3- 4 clicks on the brightness icon of Power point.I increased brightness } of all the micrographs and copied the file to a CD hoping that the image } will not deteriorate there and then compare it with the one in my laptop } several weeks from now. Has this happened to anyone else? Is there } something I should have done to prevent this? } } Any suggestions or comments will be greatly appreciated. } } Cora Bucana } }
Compliments for the season to all. And for all TEM fans " May all darkness be filled with stable clear reproducible publishable results!" We are if full swing here in Gaborone hiding from the scorching 42 degree Celsius heat in the cold EM lab battling with all the weird and wonderful samples frown at us a "quick and easy to do"
Please give help in getting a cross section of Poly polysulfone membrane microdialysis filter to reveal the structure for SEM investigation. We would not like to embed the filter. All sectioning with blades, scalpels and under LN did not result in the expected crisp clean filter surface. Fracture mess and badly smeared surfaces is all we get!
Thanks in advance.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
No experience with Electros can models. , but on an XL-30 it sounds like electrical breakdown in the gas field. The "noise" is a good indication you are very close to full breakdown and arcing is imminent. Bright pulses indicate arcing. Is detector voltage too high or sample too close to the detector. From your message it sounds like gas regulation might be flaky if imaging conditions and sample are similar to what you used in the past. . Sample charging +or- may also play a role. Again no experience with an E3 though and I almost never use the hook and cone detector system you describe. Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Gordon Vrololjak {gvrdolja-at-nature.Berkeley.EDU} 01/16/03 02:42PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hello Everyone, I came across a new problem with our Electroscan E3 ESEM, that I've not been able to track down. At times, the image will become very noisy and streaky with really bright streaks across the image making it near impossible to focus. The problem goes away with time by venting and repumping the chamber, or fiddling with the detector ESD hook wire - but I'm not sure exactly what is causing it. Anyone ever seen this before? Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
Gordon, Sounds like a dirty hi-vac gauge - Cold Cathode, Penning, etc.. Follow the tips on the List Server lately about cleaning Discharge gauges.
Gary M. Easton, Pres. Scanners Corporation SEM Service/EDS/Digital Imaging Products 410-857-7633
----- Original Message ----- } From: "Gordon Vrololjak" {gvrdolja-at-nature.Berkeley.EDU} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, January 16, 2003 2:42 PM
This same phenomenon happens with my reports in Microsoft Word. In addition to images darkening, they sometimes shift to other pages and/or change size and dimension. Adding tables and text boxes to the report adds to the fun.
The software that we use is networked. Our IS department has told us that the networked software has a bug that causes these things to happen when file sizes increase, and that there is not a patch for it. So we have just have to deal with it.
Jeff Oakley
-----Original Message----- } From: Corazon D. Bucana [mailto:bucana-at-audumla.mdacc.tmc.edu] Sent: Thursday, January 16, 2003 11:11 AM To: Microscopy-at-sparc5.microscopy.com
I created a Power point presentation last November 2002 consisting of several fluorescence micrographs. The file which was rather large ( 90 Mb) was left in my laptop all this time. When I opened it again this week I find that the images are now too dark and I needed to increase brightness by 3- 4 clicks on the brightness icon of Power point.I increased brightness of all the micrographs and copied the file to a CD hoping that the image will not deteriorate there and then compare it with the one in my laptop several weeks from now. Has this happened to anyone else? Is there something I should have done to prevent this?
Any suggestions or comments will be greatly appreciated.
In principle, the beam current measured on the CUP should be the same as the current measured using a faraday cup on the sample holder (the "ABS" or absorbed current). Also, as one increases the beam current, the ratio between the two (ABS/CUP), which should be close to one, should not change. At least that's what we thought.
When we did a very simple and quick test, we found that at low currents ( {1 na) the ABS current was slightly (a few percent) lower than the CUP current. Then, as we increased current and compared the two, the ratio ABS/CUP increased to a value of one at a few na, then went above one (to about 1.02) with increasing current. In other words, the ratio wasn't always one, and wasn't constant - it varied systematically, increasing several percent over a change in current from {1 to a few hundred na.
This only becomes a problem if the analyst calibrates at one current and analyzes at another (which we sometimes do for reasons I won't go into) and if it is the CUP current that is causing this variation (which we are convinced it is). So we're trying to sort it out.
I'm looking for others on microprobes to run the same test: simply put a good faraday cup on your sample holder (you can make one with an old objective aperture mounted on top of a larger hole drilled part way into brass), then at different currents, measure the ratio of ABS/CUP. Then plot them versus CUP.
Please send results as an Excel file (if possible) directly to me (so as not to bore the rest of the list population), and I'll compile results to share with everyone later.
If you run this test, please let me know what sort of machine you are using (we have a 15-yr old JEOL 733).
In advance, thanks,
Dave -- _________________________________________________ David A. Wark Dept of Earth & Environmental Sciences 1C16 Science Center; Rensselaer Polytechnic Institute Troy, New York 12180 office: 518-276-2674 fax: 518-276-6680 http://www.rpi.edu/~warkd/wark.html
Have you tried Focussed Ion Beam (FIB)? I didn't take the time to look up the Tg of polysulfone but would expect it to be rather high. FIB has been shown useful in such samples for preparing sections or flat faces for TEM and SEM.
Good luck,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"Coetzee, Mr S. H Physics Science" To: "Listserver Microscopy (E-mail)" {COETZEES-at-mopipi.u {Microscopy-at-sparc5.microscopy.com} b.bw} cc: Subject: Cross section of Poly polysulfone membrane microdialysis filter - SEM 01/17/03 03:57 AM
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Dear All
Compliments for the season to all. And for all TEM fans " May all darkness be filled with stable clear reproducible publishable results!" We are if full swing here in Gaborone hiding from the scorching 42 degree Celsius heat in the cold EM lab battling with all the weird and wonderful samples frown at us a "quick and easy to do"
Please give help in getting a cross section of Poly polysulfone membrane microdialysis filter to reveal the structure for SEM investigation. We would not like to embed the filter. All sectioning with blades, scalpels and under LN did not result in the expected crisp clean filter surface. Fracture mess and badly smeared surfaces is all we get!
Thanks in advance.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
I'm sort of conducting a survey - how do you freeze blocks prior to cryostat sectioning? A PI in our department is writing a grant and offered to include a freezing set-up (we just use a dewar of LN2 now) - the only equipment I've come across on the web is the giant NesLab bath. Is a styrofoam cooler of LN2 with a beaker of some solvent still a reasonable way to go, or is there some spiffy gizmo we can buy?
Thanks! (Sorry about the cross-post for those of you on both lists)
Tamara
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
A Microwave Specimen Preparation Workshop sponsored by the Life Science Microscopy Facility in conjunction with Ted Pella, Inc. will be held at Purdue University (West Lafayette, IN) on March 20-22, 2003. The workshop will be conducted by Mr. Rick Giberson and will be structured as follows:
Thursday, March 20: Introduction to microwave preparation theory. Preparation of samples Friday, March 21: Use of the microwave for Immunocytochemical localization for LM & EM - theory.... practical for TEM Saturday, March 22: Evaluating samples, discussion of decalcification, and other potential uses for scientific microwave ovens.
The course will be limited to 10 hands-on participants. Course emphasis may change depending on participant's requests.
The cost for the workshop will be $350. This fee includes registration and meals (3 breakfasts, 3 lunches, 2 dinners). Participants will be responsible for their transportation and lodging.
Please contact us if you would like additional information or a registration form.
Thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building West Lafayette, IN 47907
Hi Leslie, I you were going to use the taenia coli from a mouse, then you should consider confocal laser scanning microscopy with an inverted LM. Perhaps the same would be true if you stripped the muscularis from the mouse esophagus proximal to the oral cavity. In the former you would get only smooth muscle and in the latter a transition from skeletal to smooth(I think that's true for the mouse, though I have no sections by which to check.) The simplest, multi-cellular 'smooth muscle-like' multicellular contractile system in the mammal that I have found is the so-called myoid (or peritubular) layer on the surface of the seminiferous tubule. It is aneural, thus its contractions are myogenic with potential contributions from neighbors since they are coordinated and produce a traveling wave that can be easily seen with a LM. Finally, it responds strongly to oxytocin and related octapeptides (how many amino acids?) which often leads, in a Petrie dish, to contractual extrusion of tubular contents leaving the empty peritubular cylinder. We used Hank's MEM as a medium as well as PBS, and we did simple things like varying the temp to demonstrate the effect of T on contraction rates, etc. With an isolated seminiferous tubule from a rat (they're easier to extract than those in a mouse testis), you could run some patch-clamp type electrophysiology and couple that with caged-fluorescence. Simple but sophisticated, non-neural, monolayer myo-cellular system from which you can visualize intracellular fluorescence while you are treating the cylinder as a wire!
If you use {seminiferous tubule contractility} as a search string on PubMed you should find plenty of information to demonstrate how far behind I am in the literature of the subject.
Hope this helps and good luck,
Fred Monson Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail to Geology West Chester University of Pennsylvania Schmucker II Science Center, Room SS024 South Church Street and Rosedale Avenue West Chester, PA, 19383 Phone: 610-738-0437 eMail: fmonson-at-wcupa.edu An FEI (Quanta 400 and Technai 12), Oxford INCA Energy 400, and Olympus FV-300 Shop.
-----Original Message----- } From: gtg187h-at-mail.gatech.edu [mailto:gtg187h-at-mail.gatech.edu] Sent: Thursday, January 16, 2003 8:00 PM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (gtg187h-at-mail.gatech.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, January 16, 2003 at 10:28:03 ---------------------------------------------------------------------------
Email: gtg187h-at-mail.gatech.edu Name: Leslie Smith
Organization: Georgia Instituite of Technology
Education: Undergraduate College
Location: Atlanta, GA, USA
Question: My BioMedical Engineeering class is working ona problem dealing with Electonic Muscle Stimulation. we were wonderng what types of microscopy would be best for viewing muscles. Thanks so much Leslie Smith
I've been digging through the archives looking at printer comments, and find no mention of the new (?) Epson 2200 inkjet. Has anyone used or tested the print quality of this printer? Especially as regards print resolution?
Phil -- Philip Oshel Supervisor, BBPIC microscopy facility Department of Animal Sciences University of Wisconsin 1675 Observatory Drive Madison, WI 53706 - 1284 voice: (608) 263-4162 fax: (608) 262-5157 (dept. fax)
Where can I get the Photoshop plug-in for our Sony DKC 5000 "Catseye" digital camera? It got lost while the computer was upgraded. Thanks.
Chris A. Edwards, Mgr. Microscopy and Image-analysis Laboratory Dept. Cell & Developmental Biology The University of Michigan, School of Medicine 4747 Medical Sciences II Bldg. Ann Arbor, Michigan 48109-0616 Office: 734-936-4912 Lab: 734-763-1170 FAX: 734-763-1166
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (keithi-at-svusd.k12.ca.us) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Saturday, January 18, 2003 at 01:25:37 ---------------------------------------------------------------------------
Email: keithi-at-svusd.k12.ca.us Name: Ian Keith
Organization: La Paz Intermediate School
Education: 6-8th Grade Middle School
Location: Mission Viejo CA US
Question: I am trying to find black and white drawings of some of the most common protozoans found in pond water. I have identified several single and multicellular organisms but I don't know what they are. I would like to make a guide that shows these protozoans so my students can properly identify them. Do you know where I can find such an item??
} In principle, the beam current measured on the CUP should be the same } as the current measured using a faraday cup on the sample holder (the } "ABS" or absorbed current). Also, as one increases the beam current, } the ratio between the two (ABS/CUP), which should be close to one, } should not change. At least that's what we thought. } } When we did a very simple and quick test, we found that at low } currents ( {1 na) the ABS current was slightly (a few percent) lower } than the CUP current. Then, as we increased current and compared the } two, the ratio ABS/CUP increased to a value of one at a few na, then } went above one (to about 1.02) with increasing current. In other } words, the ratio wasn't always one, and wasn't constant - it varied } systematically, increasing several percent over a change in current } from {1 to a few hundred na.
Are both of these methods using the identical circuitry and picoAmp meter. I daresay not, and small differences could be due to some impedance or resistance differences(?) In any case, conclusions from such a study would be dependent these factors and the variety between manufacturers.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
} I've been digging through the archives looking at printer comments, } and find no mention of the new (?) Epson 2200 inkjet. Has anyone used } or tested the print quality of this printer? Especially as regards } print resolution?
Relative to my experience with an Epson 1280, I believe you'll find the printer's resolution everything you require. However, there will probably be gray neutrality issues because it will print grayscale with the colored inks (if you choose "black ink only", the resolution will be worse by an approximate factor of 4). The only other issue will probably be speed (altho this printer is probably faster than my 1280).
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
I'm lookiing to get a couple of complete EDX / EDS systems suitable for an old Cambridge Instruments S-250 Mk III, and an Electroscan 2020 ESEM. Does anyone have such , or know of someone who does, and would be willing to donate to me??? Would of course pay for package and shipping, or if in the UK can collect. Anything considered,
Cheers,
Jim
Jim Buckman Pet Eng Heriot-Watt University Edinburgh Scotland
Can you also share your comments on the highest-resolution dye-sublimation printers? In particular competition to Codonics (so far unmatched). Vendors welcome !
At 04:56 PM 1/17/03 -0600 Friday, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
And here in Ottawa, Canada, we have a non-scorching -30C at the moment, so maybe if we could average things out, we'd both be better off!
A key question, is the filter large enough to cut off a piece a few mm in cross-section and a cm or so in length? If so, then microtomy (diamond knife sectioning, found perhaps in Biology or the medical school or a major hospital) could do you just fine. If large enough, but smearing when sectioned at room temperature (hopefully less than 42C!), you would require what is called cryoultramicrotomy, ie a microtome outfitted with a liquid nitrogen attachment to cool the specimen and render it more brittle, hence easier to section smoothly. As to Gary Gaugler's suggestion of using FIB, I agree that is probably the best solution, but I suspect that, at several hundred $k US, there are not too many FIBs in Botswana.
Tom
Dr. Tom Malis Scientist Advisor / Conseiller Scientifique CANMET-Mineral Technology Branch / Direction de la technologie minerale Natural Resources Canada / Ressources naturelles Canada 555 Booth St. / 555 rue Booth, Ottawa, Ontario K1A 0G1 Tel.: (613) 995-7358 FAX: (613) 947-6606 malis-at-nrcan.gc.ca
-----Original Message----- } From: Coetzee, Mr S. H Physics Science To: Listserver Microscopy (E-mail) Sent: 1/17/2003 4:57 AM
Dear All
Compliments for the season to all. And for all TEM fans " May all darkness be filled with stable clear reproducible publishable results!" We are if full swing here in Gaborone hiding from the scorching 42 degree Celsius heat in the cold EM lab battling with all the weird and wonderful samples frown at us a "quick and easy to do"
Please give help in getting a cross section of Poly polysulfone membrane microdialysis filter to reveal the structure for SEM investigation. We would not like to embed the filter. All sectioning with blades, scalpels and under LN did not result in the expected crisp clean filter surface. Fracture mess and badly smeared surfaces is all we get!
Thanks in advance.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
I don't know, Wil. Sounds like a curse to me (you know, "May you get what you wish for."). I think I'd prefer one big leak to 50 small leaks. Same net effect vacuum-wise, but usually easier to find and fix ;-)
Ken Converse owner Quality Images Third party SEM service Delta, PA
I have always made my Reynold's lead citrate using what I believe is the standard method (see below) and never tested the pH. Recently I checked it and it was just over 13. Some sources say it should be 12 +/- 0.1. What's the consensus - do people check? How do you adjust it if you find it is high - restart and use less NaOH? Thanks, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
True about the cost of a FIB. But there are probably places that would be suitable for outsourcing at a few hundred dollars. They are all over the place here in California.
gary g.
At 11:05 AM 1/18/2003, you wrote:
} And here in Ottawa, Canada, we have a non-scorching -30C at the moment, so } maybe if we could average things out, we'd both be better off! } } A key question, is the filter large enough to cut off a piece a few mm in } cross-section and a cm or so in length? If so, then microtomy (diamond } knife sectioning, found perhaps in Biology or the medical school or a major } hospital) could do you just fine. If large enough, but smearing when } sectioned at room temperature (hopefully less than 42C!), you would require } what is called cryoultramicrotomy, ie a microtome outfitted with a liquid } nitrogen attachment to cool the specimen and render it more brittle, hence } easier to section smoothly. As to Gary Gaugler's suggestion of using FIB, I } agree that is probably the best solution, but I suspect that, at several } hundred $k US, there are not too many FIBs in Botswana. } } Tom } } Dr. Tom Malis } Scientist Advisor / Conseiller Scientifique } CANMET-Mineral Technology Branch / Direction de la technologie minerale } Natural Resources Canada / Ressources naturelles Canada } 555 Booth St. / 555 rue Booth, Ottawa, Ontario K1A 0G1 } Tel.: (613) 995-7358 FAX: (613) 947-6606 } malis-at-nrcan.gc.ca } } } } -----Original Message----- } } From: Coetzee, Mr S. H Physics Science } To: Listserver Microscopy (E-mail) } Sent: 1/17/2003 4:57 AM } Subject: Cross section of Poly polysulfone membrane microdialysis filter - } SEM } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
keithi-at-svusd.k12.ca.us (by way of Ask-A-Microscopist) wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good luck, Nelson Conti
164 Ferne Court Palo Alto, CA 94306 Email: [ncontiSFSU-at-netscape.net]
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I am going to make cross section sample of the interface between carbon nanotube film (a few hundred micrometer thick) and Si substrate for TEM. I tried the standard method to make cross section (gluing the samples with M-bond, mechanical polishing and ion milling), but the film turned to be peeled off from substrate during polishing or ion milling, due to the large thickness of film and weak bonding between the film and substrate. Any ideas and suggestions from your experience?
Best regards,
Yiming ------------------------------------ Dr. Yiming Yao
Microscopy and Microanalysis Department of Experimental Physics Chalmers University of Technology SE-41296, Göteborg Sweden
} Email: keithi-at-svusd.k12.ca.us } Name: Ian Keith } } Organization: La Paz Intermediate School } } Education: 6-8th Grade Middle School } } Location: Mission Viejo CA US } } Question: I am trying to find black and white drawings of some of the } most common protozoans found in pond water. I have identified several } single and multicellular organisms but I don't know what they are. I } would like to make a guide that shows these protozoans so my students } can properly identify them. Do you know where I can find such an } item?? } } Thanks } } Ian -
"How to know the protozoa" is a classic; you should be able to locate a cheap copy on the web. It won't, of course, help with "multicellular organisms". But I'd really like you to get two books that are listed in the Project MICRO bibliography (URL below): ---------------------------- Loewer, P. 1996 Pond Water Zoo 90pp, 7x9", hardback, $16.00. ISBN 0-689-31736-0 Athenium Books for Young Readers, Simon & Schuster. Out of print; try a used book dealer. This is a well-written natural history of commonly encountered pond life, from monera to micro-arthropods. The illustrations are excellent black and white drawings, and it's written for young readers without being simplistic. Middle school.
Rainis, K.G. and Russell, B.J. 1996 Guide to Microlife 287pp, 5.5x8.5", paperback, $40.00. ISBN 0-531-11266-7 Franklin Watts, Danbury, CT. The price of this book is both unfortunate and understandable. Unfortunate, because it should be in the library of every class that studies the microlife of our environment; understandable, because almost every page has one or more excellent color light micrographs. It's a comprehensive field guide to the microworld. The authors say that the 115 microorganisms described comprise 75-90% of those that may be encountered in the "wild". The habitats described are diverse: the home, soils, plants and debris, and four aquatic environments, with detailed advice on collecting methods for each. Described organisms are equally diverse, ranging from monerans to millimeter-sized arthropods. Species descriptions include ecological information, advice on collection and culture, and frequent suggestions for further investigation. Middle school - adult. RECOMMENDED ---------------------- If you'd like to share your completed guide with MICRO (a pdf would be nice), contact me any time.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
personally i prefer the venable's modification. as for pH, the alkalinity is what makes it go into solution and keeps CO3 precipitates down. but i never check it.
I seem to recall that there is a company which refurbishes ARL EPMAs and sells them.
Am I remembering right?
Who is it?
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
The 2200, and its overseas equivalent, the 2100, is a newish (2002 some time) Epson printer with 5 coloured inks and 2 shades of black. I imagine it will be at least equivalent to the 1290/1280 in resolution, etc. Have just been looking into getting a new printer and think this is the one we'll go for. cheers, Rosemary
} } } } Listers, } } } } I've been digging through the archives looking at printer comments, } } and find no mention of the new (?) Epson 2200 inkjet. Has anyone } } used or tested the print quality of this printer? Especially as } } regards print resolution? } } } } Phil } } -- } } Philip Oshel } } Supervisor, BBPIC microscopy facility } } Department of Animal Sciences } } University of Wisconsin } } 1675 Observatory Drive } } Madison, WI 53706 - 1284 } } voice: (608) 263-4162 } } fax: (608) 262-5157 (dept. fax)
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
Can You help with some prooven instruments/vendors offering TEM preparation technique as Jet electropolishing (similar like Fischione). Also appreciate Yr opinion about use of it and reliability of different models.
regards
Krzysztof Herman ================================ LABSOFT / FEI Service ul.Ba¿ancia 45A, 02-892 Warszawa tel/fax: (0 - 22) 6449753, 6449750 kom: (0- 601) 307456 E-mail: kherman-at-labsoft.com.pl ================================
Here at Millipore, we take cross sectional images of Polysulfone and Polyethersulfone just about every day. The key to getting a good freeze fracture in LN2 is to make sure the membrane is wet first. Here is the procedure we use:
Setup: Piece of membrane 1 or 2 pair(s) of forceps 1 weighing tin with Isopropyl Alcohol (just enough to cover the bottom of the tin) 1 weighing tin with DI or RO water (just enough to cover the bottom of the tin) 1 Styrofoam coffee cup trimmed down to make it about 2-3 inches high (5-8cm) LN2 Paper towel single edge razor blade
1. Cut a 5mm by 2cm portion of membrane from the sample. 2. Submerge membrane in Isopropyl Alcohol for approximately 1 minute to make sure the membrane is fully wet with IPA. 3. Submerge the sample in water. During this step, the sample will want to "jump around" in the water, so it may be necessary to hold it under the water with the forceps. I also find that dipping it repeatedly back in the IPA then back in the water helps with the exchange. Remember, there is TONS of surface area we're dealing with! 4. Once the piece of membrane appears to wet out with the water, pat the surfaces dry with a paper towel. Make sure to remove water from the forceps as well. 5. Grasp the sample around one third of the way up with the forceps and submerge into LN2 until the membrane and forceps are chilled. (the major bubbling around the forceps will stop. This takes about 10-20 seconds) 6. On this step, timing is key. With the forceps in your right hand, bring the index finger of your left hand close to the top of the cup. As soon as you remove the membrane from the LN2, apply pressure to the top of the strip of membrane with your left index finger until the sample fractures. This will yield the cleanest fracture. 7. You can now trim the fractured surface from the rest of the strip to about 1mm and mount it on the stub. Two-sided carbon tape works best for this.
Alternate fracture method: If you find the "finger" method to not work well for you, you can use 2 pairs of forceps to hold the strip in the LN2. When it is all frozen and still in the LN2, pull one pair of forceps towards you, and one away from you with a bit of a snap, and the membrane should break. The caveat with this method is that there is less control of where the break will occur, and sometimes it will break at the edge of the forceps, resulting in some smearing of the exposed surface.
I hope this is helpful to you, and provides an inexpensive way for you to achieve your desired results. If you would like to contact me further regarding this procedure, please feel free to do so!
Sincerely,
David Bell Scientist Electron Microscopy Lab Millipore Corporation 80 Ashby Road Bedford, MA 01730 (781) 533-2108
"Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} 01/17/2003 04:57 AM
To: "Listserver Microscopy (E-mail)" {Microscopy-at-sparc5.microscopy.com} cc: Subject: Cross section of Poly polysulfone membrane microdialysis filter - SEM
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear All
Compliments for the season to all. And for all TEM fans " May all darkness be filled with stable clear reproducible publishable results!" We are if full swing here in Gaborone hiding from the scorching 42 degree Celsius heat in the cold EM lab battling with all the weird and wonderful samples frown at us a "quick and easy to do"
Please give help in getting a cross section of Poly polysulfone membrane microdialysis filter to reveal the structure for SEM investigation. We would not like to embed the filter. All sectioning with blades, scalpels and under LN did not result in the expected crisp clean filter surface. Fracture mess and badly smeared surfaces is all we get!
Thanks in advance.
Mr S. H. Coetzee Electron Microscope Unit University of Botswana Private Bag 0022 Gaborone Botswana
I would like to mount a standard for SEM calibration, and was wondering what the best method was to do so? It appears my predesessor simply 'glued' the standard to a metal plate that was inserted into the sample holder. But it seems to me that I should use some sort of conductive adhesive? Thanks,
Jeff Roth
-- ================================================== Jeffrey Roth Science Technician James Center 205 phone: 717.245.1109 Department of Geology cell: 717.579.0644 Dickinson College fax: 717.245.1971 Carlisle, PA 17013 email: rothj-at-dickinson.edu ==================================================
I, too, have had significant problems working with images imported into Microsoft Word. However, my software is located on the PC hard drive. The biggest problem that I have encountered with images in Microsoft Word occurs when annotating images. After the image is imported into Word, annotation may be done in two ways (to my knowledge): (1) Text, arrows, etc. may be simply superimposed over the images. The problem with this approach is that the annotations are not linked to the image and may not remain superimposed on the image if the image moves. (2) Annotations can also be linked (probably not the best choice of words) to the image by double-clicking on the image to open the image field, adding the annotation, then closing the image field. These annotations are permanent unless intentionally moved or deleted.
The problems occur when one implements the second option. Comparing images before and after annotation, I found that the annotated images often sustained substantial changes in gray or color levels. Case in point, EDS maps were so badly affected that the color key was no longer correct.
My solutions follow: (1) Annotate images in Adobe Photoshop before importing into Word. Note that the effects of lossy compression on annotations (blurred edges) may be pronounced. (2) Use Microsoft PowerPoint for image presentation. I have encountered no problems with image files in PowerPoint.
Good luck to you in your endeavors.
Cheers,
"The opinions expressed are those of Gary M. Brown and do not represent the opinions of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
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This same phenomenon happens with my reports in Microsoft Word. In addition to images darkening, they sometimes shift to other pages and/or change size and dimension. Adding tables and text boxes to the report adds to the fun.
The software that we use is networked. Our IS department has told us that the networked software has a bug that causes these things to happen when file sizes increase, and that there is not a patch for it. So we have just have to deal with it.
Jeff Oakley
-----Original Message----- } From: Corazon D. Bucana [mailto:bucana-at-audumla.mdacc.tmc.edu] Sent: Thursday, January 16, 2003 11:11 AM To: Microscopy-at-sparc5.microscopy.com
I created a Power point presentation last November 2002 consisting of several fluorescence micrographs. The file which was rather large ( 90 Mb)
was left in my laptop all this time. When I opened it again this week I find that the images are now too dark and I needed to increase brightness by 3- 4 clicks on the brightness icon of Power point.I increased brightness
of all the micrographs and copied the file to a CD hoping that the image will not deteriorate there and then compare it with the one in my laptop several weeks from now. Has this happened to anyone else? Is there something I should have done to prevent this?
Any suggestions or comments will be greatly appreciated.
Hi all! The Limnology people here gave me bottles of riverine water samples + flocs. They would like SEM images of their natural riverine flocs. Do you have suggestions on how to process these floc samples? We're interested in looking at the overall floc morphology and microbial cells as well. We're also interested in looking at their inorganic matter composition.
Thanks.
Lizette Tuason Univ. of Northern British Columbia 3333 University Way Prince George, BC V2N 4Z9 Canada Phone (250) 960-5677
I would like to mount a standard for SEM calibration, and was wondering what the best method was to do so? It appears my predesessor simply 'glued' the standard to a specialized metal plate that was inserted into the sample holder. But it seems to me that I should use some sort of conductive adhesive? Thanks,
Jeff Roth
======================== Jeffrey Roth Science Technician Department of Geology Dickinson College Carlisle, PA 17013 rothj-at-dickinson.edu ========================
If you purchase a Geller MRS standard, they can be pre-mounted according to your stage's needs. These standards are for magnification and resolution cal. Mostly mag. These standards can be purchased with ISO/NIST certification. Other standards from Pella, SPI, et. al. are usually not mounted. But I suspect that you could get them mounted. There are spheres, Gold on Tin and Tin balls. These are good for checking stig and rez. Baseline at purchase and re-check periodically to see if rez or stig has shifted appreciably. Then check apertures and/or column liner.
If you buy standards, I'd recommend storing them under vacuum when not being used.
gary g.
At 05:56 AM 1/20/2003, you wrote:
} I would like to mount a standard for SEM calibration, and was wondering } what the best method was to do so? It appears my predesessor simply } 'glued' the standard to a metal plate that was inserted into the sample } holder. But it seems to me that I should use some sort of conductive } adhesive? Thanks, } } Jeff Roth } } -- } ================================================== } Jeffrey Roth Science Technician } James Center 205 phone: 717.245.1109 } Department of Geology cell: 717.579.0644 } Dickinson College fax: 717.245.1971 } Carlisle, PA 17013 email: rothj-at-dickinson.edu } ================================================== } } }
The Fuji Pictrograph 3500 blows away the Codonics and costs less, although we have had two blown circuit boards in ours.
John Mardinly Intel
-----Original Message----- } From: Marek Malecki, M.D., Ph.D., Professor [mailto:MMalecki-at-wisc.edu] Sent: Saturday, January 18, 2003 11:05 AM To: Microscopy-at-sparc5.microscopy.com
Can you also share your comments on the highest-resolution dye-sublimation printers? In particular competition to Codonics (so far unmatched). Vendors welcome !
At 04:56 PM 1/17/03 -0600 Friday, you wrote: } ----------------------------------------------------------------------- - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I haven't been able to find any laser printer sheets of appropriately sized light microscope slide labels.
ie: 7/8" wide X 6/8" high.
As a result, we have always been forced to use rolls, and type out each label tediously on a typewriter.
I was just wondering if anyone knows of any slicker system for labelling microscope slide labels. Even upstairs with the thousands of slide labels that they make, they have to tediously type them all out 1 by 1 on the typewriter.
I have found for micrographs and negative envelopes, that a combination of Microsoft Word mail merged with an appropriate Excel data base does a nice job using Avery 5160 address labels, but unfortunately, I haven't as yet been able to find a properly sized label for microscope slides, or even one that I could cut nicely. :-(
I was just wondering how other people tackled this problem. Do you all do it the tedious way too? Here is a marketing opportunity for some ambitious person who wants to make quick money, by solving a problem in the workplace.
John, Did you mean the "Fuji Pictrography 3500"? http://www.electroimage.com/fp3k.htm -Mike
"Mardinly, John" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } The Fuji Pictrograph 3500 blows away the Codonics and costs less, } although we have had two blown circuit boards in ours. } } John Mardinly } Intel } } -----Original Message----- } } From: Marek Malecki, M.D., Ph.D., Professor [mailto:MMalecki-at-wisc.edu] } Sent: Saturday, January 18, 2003 11:05 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: the highest-resolution dye-sublimation printers WAS: Re: } printer hunt } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Can you also share your comments on the highest-resolution } dye-sublimation printers? } In particular competition to Codonics (so far unmatched). } Vendors welcome ! } } At 04:56 PM 1/17/03 -0600 Friday, you wrote: } } ----------------------------------------------------------------------- } - } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------
} Nelson Conti wrote: } } } Dear Ian: } } One guide that I am aware of is titled "The Illustrated Guide to the Protozoa" published by the Society of Protozoologists (URL:http://www.uga.protozoa.edu) (pretty sure). I am a member of that Society which is how I know about such a guide. I have no idea how much such a guide would cost, but perhaps if you tried their site, there should be information about the guide I've suggested. } } The correct URL is www.uga.edu/~protozoa . } } } ______________________________________________________________________ } } Michael Nesson, Ph.D. Department of Biochemistry & Biophysics } 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 } (541)737-5245 FAX:(541)737-0481 nessonm-at-onid.orst.edu
-- _______________________________________________________________________ Michael Nesson, Ph.D. Department of Biochemistry & Biophysics 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 (541)737-5245 FAX:(541)737-0481 nessonm-at-onid.orst.edu
Yes. John Mardinly Phone: 408-765-2346 Pager: 877-277-1182
-----Original Message----- } From: Michael O'Keefe [mailto:MAOKeefe-at-lbl.gov] Sent: Monday, January 20, 2003 12:40 PM To: Mardinly, John Cc: Marek Malecki, M.D., Ph.D., Professor; Microscopy-at-sparc5.microscopy.com
Help Listservers,
I need to explain, justify, etc., to colleagues (who have no microscopy/microtomy experience or background) reasons why you should isloate TEM rooms from SEM rooms from microtome rooms? In other words, why you should avoid putting a TEM, an SEM and a cryoultramicrotome all in the same room. Thanks
Fred A. Hayes Analyst Polymer Microscopy Collins and Aikman IntelliMold Systems 4651 Platt Lane Ann Arbor, MI 48108 734-477-7029 direct 734-477-9214 fax 734-477-9212 office www.IntelliMold.net www.collinsaikman.com fred.hayes-at-colaik.com
In my last posting about the pH of Reynold's lead citrate, I didn't give the exact method we use. Here it is:
1.33 gm Pb(NO3)2 1.76 gm Na3 citrate -2H20 30 ml dH2O (all water used for this prep is freshly boiled and cooled to get rid of CO2) shake vigorously let stand 30 min Add 8 ml of freshly made 1 N NaOH dilute to 50 ml total volume.
Reynold's says the pH was "routinely found to be 12.0 +/- 0.1". The only real difference between my method and Reynold's is that he used 1 N NaOH prepared by diluting a commercial 10 N carbonate free stock. I make my NaOH from pellets immediately before use. Assuming I accurately weigh it, any error would presumably result from some water absorption by the pellets so the error would be to decrease the NaOH concentration. But the pH we get implies too much NaOH.
We got a pH of ~13.5 (with good quality pH paper - i don't want to use my pH meter for this). We cut the volume of NaOH to 7 ml and got a pH of ~12.5 or slightly less (pH paper has 0.5 unit steps). Reynold's notes that the staining intensity decreases with higher pH
My question remains - do people measure the pH of their Reynolds? do they get 12? I assume you can't adjust the pH with HCl but I am open to correction. I fully understand the concept of emphirecally testing it and seeing the results. I am not sure if my results are the best and furthermore, I would like to have a method that is consistent and that I understand so I don't need to re-invent this wheel each time. I am aware of the other lead formulations but am interested in people's experience with Reynolds. I may switch but I think switching everytime a problem comes up without attempting to understand the problem is poor science. Thanks for any comments. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I was wondering if anyone could help me with information about the proceedings for the 1993 and 1994 (51st and 52nd, respectively) Annual MSA Meetings. I am looking for the publishers for each of these years. Was it San Francisco Press or Jones & Begell? Also, I am looking for the editors for the 1993 year.
If the standard was coated after it was glued on, the coating may provide sufficient conductive pathway. If it works, it was done OK. I often mount things with Duco cement but use silver paint on edges to make sure I have a good electrical connection after coating. A conductive adhesive is another way to go, for sure.
Ron L
-----Original Message----- } From: Jeff Roth [mailto:rothj-at-dickinson.edu] Sent: Monday, January 20, 2003 1:31 PM To: Microscopy-at-sparc5.microscopy.com
I would like to mount a standard for SEM calibration, and was wondering what the best method was to do so? It appears my predesessor simply 'glued' the standard to a specialized metal plate that was inserted into
the sample holder. But it seems to me that I should use some sort of conductive adhesive? Thanks,
Jeff Roth
======================== Jeffrey Roth Science Technician Department of Geology Dickinson College Carlisle, PA 17013 rothj-at-dickinson.edu ========================
Ian, In addition to numerous sites accessible via google, I've used earlier versions of the following for many years and the current version is available at amazon.com.
Pond Life: A Guide to Common Plants and Animals of North American Ponds and Lakes (Golden Guide) by George Kell Reid, Herbert S. Zim (Editor), George S. Fichter (Editor), Jonathan P. Latimer, Karen Stray Nolting, John L. Brooks, Sally D. Kaicher (Illustrator), Tom Dolan (Illustrator) Paperback: 160 pages ; Dimensions (in inches): 0.34 x 5.94 x 3.93 Publisher: St. Martin's Press; ; Revised and Updated edition (April 2001) ISBN: 1582381305 $6.95 Rosemary
I was wondering if anyone could help me with information about the proceedings for the 1993 and 1994 (51st and 52nd, respectively) Annual MSA Meetings. I am looking for the publishers for each of these years. Also, I am looking for the editors for the 1993 year.
Check out the Struers Tenupol. Expensive, but the Hasselblad of Jet electropolishing. John Mardinly
-----Original Message----- } From: Krzysztof Herman [mailto:kherman-at-labsoft.com.pl] Sent: Monday, January 20, 2003 4:57 AM To: MSA
Hi list
Can You help with some prooven instruments/vendors offering TEM preparation technique as Jet electropolishing (similar like Fischione). Also appreciate Yr opinion about use of it and reliability of different models.
regards
Krzysztof Herman ================================ LABSOFT / FEI Service ul.Ba¿ancia 45A, 02-892 Warszawa tel/fax: (0 - 22) 6449753, 6449750 kom: (0- 601) 307456 E-mail: kherman-at-labsoft.com.pl ================================
A TEM Specimen Preparation Course Will be offered March 12-14, 2003 at the Materials Characterization Facility at the University of Central Florida, Orlando, FL USA
A review of all techniques will be given, with primary emphasis on tripod polishing, ion milling, and all latest FIB techniques
Instructors: Ron Anderson, Microscopy Today Fred Stevie, NC State Lucille Giannuzzi, UCF Brian Kempshall, UCF
For more information contact Lucille Giannuzzi, lag-at-mail.ucf.edu
******************************************************************* Lucille A. Giannuzzi, Ph.D.
Associate Professor Mechanical, Materials, and Aerospace Engineering University of Central Florida 4000 Central Florida Blvd. PO Box 162450 Orlando, FL 32816-2450 email lag-at-mail.ucf.edu phone 407-823-5770 fax 407-823-0208 -------------------------------------------------------------------- "Good judgement comes from experience.
Experience comes from making bad judgement."
Mark Twain ********************************************************************
Rosemary, The two shades of black are for use with different papers. One black for ordinary papers, the other for glossy photo papers. You can get details on the Epson web site. Damian
} The 2200, and its overseas equivalent, the 2100, is a newish (2002 some } time) Epson printer with 5 coloured inks and 2 shades of black. I imagine } it will be at least equivalent to the 1290/1280 in resolution, etc. Have } just been looking into getting a new printer and think this is the one } we'll go for. } cheers, } Rosemary
Hi all We have a client with an old JEOL 1200 TEM that was modified by the factory to have two diffusion pumps as the vacuum system. We now have a problem that the EPROMs that control the vacuum system have gone faulty and are have a real challenge trying to get some replacements. Also the manuals are all for systems with either an Ion pump or a Turbo pump as a second pump for the gun and column. This means we are really having fun trying to work out the logics.
Can anyone help us out with a set of EPROMS, the software for the EPROMS and or manuals specific for the double diffusion pump version of the JEOL 1200?
Thanks
Luc Harmsen ANASPEC South Africa www.anaspec.co.za Tel: +27 11 794 8340 Fax: +27 11 794 8349 P.O. Box 2561 Honeydew 2040 Gauteng, South Africa
This is to inform you that the Call for Abstracts for the forthcoming International Meeting on Applied Physics (APHYS-2003), to be held in Badajoz (Spain), during October 14-18th 2003, is now opened.
Website www.formatex.org/aphys2003/aphys2003.htm
Abstracts submission deadline: March 24th 2003
As you can see, Imaging Techniques and Applied Microscopy will have very important weight in the scientific Program of APHYS-2003. Some of topics to be covered will be:
- Surfaces, Interfaces and Colloids - Imaging Techniques, Microscopy - Nano-sciences and Technologies - Materials Science - Biomedical Engineering and Biomaterials Science&Engineering - Computational Physics
In addition to the regular Scientific Program, several International Workshops will be held as pre-conference events. The following two will be of high interest for the Microscopy community:
1. Workshop on Modern Applied Microscopy in Molecular and Cell Biophysics Research(please visit the website for details)
2. Pre-conference Workshop: International Interdisciplinar Workshop on Bioengineered Non-crystalline Solids
For all the information regarding the conference, please visit www.formatex.org/aphys2003/aphys2003.htm
Potencial reviewers are kindly asked to write us with personal data, field(s) of expertise and a list of publication. The official list of reviewers will be also included in the conference publications.
Proceedings will be published within several international journals, depending on each paper topic:
- Journal of Microscopy - Journal of Non-crystalline Solids - Applied Surface Science - Physica Scripta
For issues regarding Commercial Exhibition and Sponsorship, please refer to the Conference website or contact Ines Solo de Zaldivar (secretariat-at-formatex.org )
Thank you in advance and please contact us for any suggestion or question.
Antonio Mendez-Vilas APHYS-2003 Co-ordinator secretariat-at-formatex.org www.formatex.org/aphys2003/aphys2003.htm
} Can anyone suggest a UK supplier of Polyvinyl Alcohol } at 24-32 centipoise viscosity?
Another possibility is mixing PV-acetate with ethanol. I believe my recipe is 20% by weight, which will lay down, dry and become a heat-sensitive "glue" for relatively large samples. I'd make the mixture 10% for very small particles.
cheerios ... shAf :o) Avalon Peninsula, Newfoundland www.micro-investigations.com (in progress)
Yes, pH the stain to around 12 +/- 0.1 You do get erratic staining with a higher pH.
Best of Luck,
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-8806 calomeni-1-at-medctr.osu.edu
} } } Tom Phillips {phillipst-at-missouri.edu} 01/18/03 05:47PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I have always made my Reynold's lead citrate using what I believe is the standard method (see below) and never tested the pH. Recently I checked it and it was just over 13. Some sources say it should be 12 +/- 0.1. What's the consensus - do people check? How do you adjust it if you find it is high - restart and use less NaOH? Thanks, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
I am a happy user of commercial CO2-free, titrated NaOH. I buy mine from Electron Microscopy Sciences [in which I have no commercial interest, wish I had :)]. It is inexpensive, something like $10 a bottle. This *is* the consistent method, as long as you keep using the same water of course. When I was making Reynolds from pellets, sometimes I would get stain that peppers easily and sometimes the stain would come out too weak. I think NaOH stock is the key, once you have good water.
Paper shows pH of my stain about 12. I never measured with pH meter but I know there are electrodes which would allow you to do that, I once asked an Orion tech. I do remember seeing in publications that it is important that the pH be 12+/-0.1, and if I was running a teaching lab, I would probably get a proper electrode and measure and maybe experiment adjusting if the pH turned out different from 12+/-0.1. Here, however, I am content with the fact that my Reynolds stains reproducibly well.
As for the other lead stain recipes, I often use the Venable-Cogeshall [sp?] instead of Reynolds. It does not make as "sweet" a picture 20k and above as Reynolds but does seem to give more contrast. I use it for no more than 2 days, then make fresh the next time (have pre-weighed LC aliquotes sitting in 15 ml Falcons and just add water and NaOH when the time comes). Again, keeping the same water and NaOH stock makes it highly consistent.
All the best, Vlad
} In my last posting about the pH of Reynold's lead citrate, I didn't } give the exact method we use. Here it is: } } 1.33 gm Pb(NO3)2 } 1.76 gm Na3 citrate -2H20 } 30 ml dH2O (all water used for this prep is freshly boiled and } cooled to get rid of CO2) } shake vigorously let stand 30 min } Add 8 ml of freshly made 1 N NaOH } dilute to 50 ml total volume. } } Reynold's says the pH was "routinely found to be 12.0 +/- 0.1". The } only real difference between my method and Reynold's is that he used } 1 N NaOH prepared by diluting a commercial 10 N carbonate free } stock. I make my NaOH from pellets immediately before use. } Assuming I accurately weigh it, any error would presumably result } from some water absorption by the pellets so the error would be to } decrease the NaOH concentration. But the pH we get implies too much } NaOH. } } We got a pH of ~13.5 (with good quality pH paper - i don't want to } use my pH meter for this). We cut the volume of NaOH to 7 ml and } got a pH of ~12.5 or slightly less (pH paper has 0.5 unit steps). } Reynold's notes that the staining intensity decreases with higher pH } } } } My question remains - do people measure the pH of their Reynolds? } do they get 12? I assume you can't adjust the pH with HCl but I am } open to correction. I fully understand the concept of emphirecally } testing it and seeing the results. I am not sure if my results are } the best and furthermore, I would like to have a method that is } consistent and that I understand so I don't need to re-invent this } wheel each time. I am aware of the other lead formulations but am } interested in people's experience with Reynolds. I may switch but I } think switching everytime a problem comes up without attempting to } understand the problem is poor science. Thanks for any comments. Tom } } } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu
-- -------------------------------------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
Fred, Get you hands on a copy of the book in the Glauert series :Design of the Electron Microscope Laboratory (or something very close to that name, I don't have it in from of me). I have found it of enormous value, although it remains to be seen if I too will succeed in a similar argument. good luck, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
Post-Doctoral Research Associate Stevens Institute of Technology
There is a post-doctoral opportunity within the Department of Chemical, Biochemical, and Materials Engineering at the Stevens Institute of Technology to develop and apply advanced techniques of transmission electron microscopy to quantitatively study the morphology of both dry and hydrated synthetic and natural polymers.
The ideal candidate will have experience in electron optics, electron energy-loss spectroscopy, and cryo-TEM. Acceptable candidates will at least have: (1) experience with transmission electron microscopy, either
from a physical or biological sciences perspective; (2) a willingness to
develop a leadership role in problems associated with the nanoscale morphology of hydrated polymeric biomaterials; and (3) good communication skills.
This is an NIH-funded position associated with a new inter-institutional
NCRR on Polymeric Biomaterials. This NCRR is associated with the New Jersey Center for Biomaterials and involves core laboratories at Rutgers, NJIT, and Stevens. A multi-year appointment for this position is anticipated.
The Stevens Institute of Technology is a small private university concentrated on engineering, science, and technology management. Stevens is located in very close proximity to New York City. The Stevens electron-optics laboratory contains a Philips CM20 FEG TEM/STEM,
a Philips CM30 SuperTwin TEM, and a LEO 982 FEG SEM. The CM20 FEG TEM/STEM is equipped with a Gatan Enfina ccd PEELS system and a Gatan Multiscan digital camera. Both are interfaced to an Emispec Vision acquisition and control system. The facility is fully equipped with cryomicrotomy and cryo-transfer capabilities to deal with frozen hydrated materials.
For further information please contact:
Professor Matthew R. Libera Dept. Chemical, Biochemical, and Materials Engineering Stevens Institute of Technology Hoboken, New Jersey 07030 ph: 201-216-5259 fax: 201-216-8306 mlibera-at-stevens-tech.edu
MS Word is such a pain because of the way it works with images. A couple of tricks I've discovered over the years.
1) Place the image within a text box, rather than directly on the page, it seems to give you much greater control over the location on the page. It also makes it much easier to add text annotations that stay with the image.
If you don't want the text box to have a border you can remove it by selecting the box outline and look for the "paint brush" icon on the Draw toolbar, then use the down arrow and select "none". If you'd like the text box to be transparent, select the text box outline and look for the "paint bucket" icon on the Draw toolbar, then use the down arrow and select "none". If you discover its hard to find the text box border once you made the edge "invisible", first select the image with a single left click (it should have the solid black resizing "handles") and then use the keyboard left or right arrow keys and the selection with move out to the textbox outline (with black bordered resizing "handles").
2) Always use the INSERT | PICTURE | FROM FILE option as opposed to copying and pasting an image into Word. I find that the images are harder to work with if I paste them in. Also, you can insert TIFF or BMP images into Word, you don't have to use JPEG.
FYI: Last known, there was inexpensive software on the web that compressed bloated Powerpoint files. I also have a freeware utility on my PC called "packword" that compresses Word 97 and 2000 files. I'm sorry I don't have the URLs, I'm writing this from home.
Best regards, Doug
} -- Original Message -- } Date: Mon, 20 Jan 2003 09:30:47 -0600 } From: "gary.m.brown-at-exxonmobil.com"-at-sparc5.microscopy.com } Subject: RE: presentation images in Microsoft Word } To: oakleyj-at-rayovac.com } Cc: Microscopy-at-sparc5.microscopy.com } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} } "Oakley, Jeff"
} } {oakleyj-at-rayovac.c To: {Microscopy-at-sparc5.microscopy.com} } } om} cc:
} } Subject: RE: presentation } images }
} } 01/17/03 07:50 AM
} }
} }
} } } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
.......................................................................... Douglas W. Cromey, M.S., Principal Research Specialist Dept. of Cell Biology & Anatomy, University of Arizona Tucson, AZ 85724-5044 USA 520-626-2824 {cromey-at-arizona.edu} .......................................................................... http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW"
Usually all microscope columns are sensitive to electromagnetic fields and should be placed on distances at least 10 feet from each other and from power lines. Additionally microscopes are sources of vibrations and acoustic noise which could degrade images of high resolution instruments.
And, sure, it is absolutely inconvenient for operators to have SEM and TEM in the same room: working TEM requires full darkness and this will give SEM operator only unnecessary eye fatigue and problems with specimen replacement. I could agree that it is possible to install couple of older (and less sensitive) SEMs in the same room, but each TEM should have separate light-tight room.
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } Help Listservers, } } I need to explain, justify, etc., to colleagues (who have no } microscopy/microtomy experience or background) reasons why you should } isloate TEM rooms from SEM rooms from microtome rooms? In } other words, why } you should avoid putting a TEM, an SEM and a } cryoultramicrotome all in the } same room. Thanks } } Fred A. Hayes } Analyst } Polymer Microscopy } Collins and Aikman } IntelliMold Systems } 4651 Platt Lane } Ann Arbor, MI 48108 } 734-477-7029 direct } 734-477-9214 fax } 734-477-9212 office } www.IntelliMold.net } www.collinsaikman.com } fred.hayes-at-colaik.com } } }
I posed this question in late December when many were on Christmas vacation so will try again. Does anyone know the original citation for double stick carbon tape used on SEM specimen holders? I think it was first used in the mid 1990's but I have been unable to locate when and who introduced it.
Dear Fred, TEM rooms must be able to be completely darkened and are the ones that must be isolated from noise, vibration, air currents, etc. Microtomes would have to be vibration isolated from the pumps on SEMs and TEMs. ----- Original Message ----- } From: "Hayes, Fred" {Fred.Hayes-at-colaik.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, January 20, 2003 3:48 PM
We have a CoStar label maker that we purchased some time ago for regular mailing labels. It takes stock of various widths and factors the appropriate pitch into its software. We just run off a ribbon of labels one wide by as many long as we need.
Warren
At 02:20 PM 1/20/03 -0600, you wrote:
} I haven't been able to find any laser printer sheets of appropriately sized } light microscope slide labels. } } ie: 7/8" wide X 6/8" high. } } As a result, we have always been forced to use rolls, and type out each } label tediously on a typewriter. } } I was just wondering if anyone knows of any slicker system for labelling } microscope slide labels. Even upstairs with the thousands of slide labels } that they make, they have to tediously type them all out 1 by 1 on the } typewriter. } } I have found for micrographs and negative envelopes, that a combination of } Microsoft Word mail merged with an appropriate Excel data base does a nice } job using Avery 5160 address labels, but unfortunately, I haven't as yet } been able to find a properly sized label for microscope slides, or even one } that I could cut nicely. :-( } } I was just wondering how other people tackled this problem. Do you all do } it the tedious way too? Here is a marketing opportunity for some ambitious } person who wants to make quick money, by solving a problem in the workplace.
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 23 Town Engineering Ames IA, 50011-3232
Listers, We are going to try to plunge freeze and freeze substitute mouse pancreas. Now I know that the most desirable way to do this would be with high pressure freezing but that is not an option at the moment. So I am hoping to get a bit of help with what we do have available.
We have done quite a bit of plunge freezing microbes and fungi into Liquid nitrogen cooled liquid propane and then substitution in ethanol, acetone, etc. with good results. Embedding resin depends on tissue...Epoxy (LX112) or HM20 lowacryl as needed.
I would like advice on using a cryo-protectant for the mouse pancreas ..thinking of using 18% glycerol or 18% propylene glycol. Is this necessary and do you recommend one over the other?
Also for substitution, I was going to use acetone + 2% osmium for standard ultrastructure and just acetone for ICC (switched to ETOH for infiltration with HM20.
I would appreciate comments on above general approach and any other information you would like to share that may enhance contrast on final sections.
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 West Lafayette, IN 47907
} It seems this man's daughter is suffering from some kind of skin ailment } and she has dug stuff out of MANY of the sores on her skin. The } doctors in her HMO have not offered any relief. Her father (from another } department on this campus) is quite determined to find out what the problem } might be.
OK, I'm not a doctor and, compared to most on this list, I know very little about microscopy, but...
when I was young I had a skin ailment that sounds somewhat like what you are describing. You provided very little information on the skin ailment itself, so I may be way off base here. What I had was a rash of boils (at one point I had 42 of them at one time on various parts of my body). I would expect the doctors at your HMO to be able to recognize boils, but perhaps they're not all that common anymore. Anyway, the eruptions had a fibrous core, sometimes more than one. This core was so cohesive that when finally removed it left a cylindrical hole in the sore. Sometimes the core would come out on its own along with gobs of pus, sometimes I would grab it with a pair of tweezers and pull it out. After removal of the core the skin eruption would subside and disappear.
Hope that helps.
Bruce Girrell
BTW - The doctor prescribed moist heat to help the boils drain and would have us wash them regularly with PhisoHex (no longer available) soap. Sometimes the doctor would lance the boils to drain them. None of this did anything to prevent recurrances. My grandfather said "You need a blood cleanser. Drink burdock root tea." So I drank that nasty stuff (no sweetening or anything else) for about two weeks. It may just be a coincidence, but I have never had another boil.
I'd suggest much earlier than the mid 1990s; I first learned SEM in 1993, and my impression then was it was old hat already. Perhaps the mid 1980s? Ben Simkin (simkin-at-egr.msu.edu) dpt. Chemical Engineering and Materials Science Michigan State University
We are experiencing artifacts in thick epon-araldite resin sections and are about at our wit's end. You can view the artifact by linking to http://biotech.missouri.edu/emc/stain_artifact.jpg and noting the dark magenta blobs in the nuclear layer of the retinal tissue in the image. We have determined that the "blobs" are associated with "dimples" in the sections, but they have never shown up in hundreds of other samples done previous to this particular project. We are virtually certain this is artifact because serial sections show the blobs in different places and in different concentrations.
What we have done so far is to try varying the section thicknesses from from 0.4 microns to 3.0 microns, mixing the aqueous toluidine blue 1:1 with varying concentrations of ethanol from 20% to absolute, diluting the stain with water, mixing fresh stain, cutting on both glass and diamond histo knives, trying different temperatures on the hot plates, trying untreated slides and slides treated with BSA, staining in the microwave at varying wattages and times, and dipping slides and sections in methanol before staining, then staining normally.
So far, the only things that have made any difference are using ethanol in the stain and dipping the slides in methanol prior to staining, but neither are consistent. They might work one time, but not the next and we still get the blobs showing up in different places (but ALWAYS in the nuclear layer only) and in different numbers.
HEELLLPPP!!!
Most grateful for ANY assistance with this one!
Randy
Randy Tindall EM Specialist Electron Microscopy Core----We're the Fun Core! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
Hi Debby: We just got our new HPF so that will teach you for leaving us! For freeze-sub without aldehydes or osmium, add 0.1% uranyl to the acetone since it will increase membrane contrast. a good paper on this is Moreira et al. J Histochem Cytochem 46(7):847-854. Leica's web site has a large compilation of freeze sub protocols. I have used cryoprotectants with fixed material (e.g., the Tokyasu method) but I haven't frozen unfixed material with cryo-protectants. I would worry about the osmotic stress and diffusion of the material. I am a little doubtful about the quality of freezing you will achieve with this approach. Tom
At 01:53 PM 1/21/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400