For changing of scientific information we are needing E-mail addresses following persons: Prof. J.-P. Chavalier, C.E.C.M.-C.N.R.S., France; Dr. A.G. Norman, Oxford University, UK; Dr. J.L. Hutchison, Oxford University, UK; Dr. W.M. Stobbs, Cambridge University, UK; Dr. C.S. Baxter, Cambridge University, UK; Dr. R.F. Broom, IBM Research Division, Zurich Research Laboratory, Sw; Prof. S.Mahajan, Carnegie Mellon University, US; Dr. I.T. Ferguson, Imperial College, London, UK; Dr. S.J. Pennicook, Oak Ridge National Laboratory, US; Dr. O. Ueda, Fujitsu Laboratories, Jupan.
Please reply on Microscopy list or directly on our E-mail addres: lemi-at-mx.iki.rssi.ru We will be grateful for any information. Sincerely Yours Prof.S.Maksimov CFPM, Electron microscopy laboratory, Moscow Institute of Electronic Technology, Russuia.
I am desperately trying to find someone who has a Durst Laborator S-45 EM enlarger that they no longer need. We are particular interested in finding the NEGA-138 carriers with glasses. Also the upper portion containing bellows and focusing devices. Any help would be very much appreciated. We have two enlargers and need to keep them in operational order. Thanks Cathy Kelloes
We used to prepare nanoparticles for TEM by simply touching a coated (carbon, formvar or collodium) grid directly on the surface of the powdered sample. We studied with this method the particle size and shape distribution of several inorganic pigments, zeolites, polymers, powdered bioinsecticides, etc. It is good to measure particle dimensions, but particle distribution is affected by preferential adsorption of particles (For example in the range of sizes 50-250 nm for iron pigments). Dagoberto Rodriguez Montreal, Qc, Canada
} i frequently have Al detected when doing EDXA on plain carbon } steel samples, even though i mount the samples using a carbon- } based adhesive. However, i do not detect it all the time. any } suggestions as to why this is occurring? } Dear Tania, Is there any Al in your 'scope (liners, etc.)? Do you see Al when you look at the background counts from a position right next to your sample? Do you suspect there may be Br in the specimen (overlap with Al-k, but with two peaks at } 10 keV)? Good luck. Yours, Bill Tivol
I am a graduate student at the Univeraity of Wisconsin-Milwaukee and I have a problem with a technique.
I embed bacteria in LR White. I did ultrastructural work and stained with UA and Pb and had no major problem with holes in the resin. The next project is to immunostain a protein in the membrane. I can use the same blocks as before and after immunostaining, there are multiple holes in most cells. I put the sections on nickel grids. NaCl for 1 min. New BSA 3% filtered for 1 min. Primary antibody for 1 hour. BSA washes. Secondary antibody IgG for 1 hour. BSA washes. NaCl wash. Water washes. The I stain with UA for 1 hour as per usual.
Help.....I'd like to get rid of the holes. I think that it is happening during this procedure. The new BSA has not eliminated them as was suggested that it might.
Thank You
Chris Brantner
cab-at-csd.uwm.edu University of Wisconsin-Milwaukee
Many years ago I used an ultra-low viscosity medium for rapid embedding of plant cells. This resin prevented plasmolysis and the tissue did not separate from the plastic. We published our findings and the recipe for this plastic (available from Polysciences) in the following article:
We often do analyses of ground up deposits which contain iron oxides. Sometimes, but not always, the EDS results will have an unreasonably large amount of iron, with not enough oxygen, even though we know from XRD that the material is iron oxide, not metallic iron. I don't know if it only occurs with a particular iron oxide or not. Has anyone experienced this or have some insight as to why this occurs?
} X-UIDL: bc714d162b18bc45931c0b85533415ac } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id } NAA17979 for dist-Microscopy; Sat, 2 Dec 1995 13:10:52 -0600 } Received: from mail06.mail.aol.com (mail06.mail.aol.com [152.163.172.108]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id NAA17976 for } {Microscopy-at-microscopy.com} ; Sat, 2 Dec 1995 13:10:51 -0600 } Received: by mail06.mail.aol.com (8.6.12/8.6.12) id OAA10021 for } Microscopy-at-microscopy.com; Sat, 2 Dec 1995 14:09:25 -0500 } From: MelanieOwl-at-aol.com } Date: Sat, 2 Dec 1995 14:09:25 -0500 } Message-ID: {951202140924_123023520-at-mail06.mail.aol.com} } To: Microscopy-at-microscopy.com } Subject: Non-stoichiometric iron oxide results with EDS } Content-Type: text } } We often do analyses of ground up deposits which contain iron oxides. } Sometimes, but not always, the EDS results will have an unreasonably large } amount of iron, with not enough oxygen, even though we know from XRD that the } material is iron oxide, not metallic iron. I don't know if it only occurs } with a particular iron oxide or not. Has anyone experienced this or have } some insight as to why this occurs? } } Thanks in advance, } Melanie Behrens } }
} X-UIDL: ae048b1c15b3a04492ad7391aa68499e } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id } OAA00310 for dist-Microscopy; Sat, 2 Dec 1995 14:20:08 -0600 } Received: from worf.uwsp.edu (worf.uwsp.edu [143.236.1.12]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id OAA00307 for } {microscopy-at-sparc5.microscopy.com} ; Sat, 2 Dec 1995 14:20:06 -0600 } Received: from macsrv1.uwsp.edu (macsrv1.uwsp.edu [143.236.1.28]) by } worf.uwsp.edu (8.6.5/8.6.5) with SMTP id OAA17716 for } {microscopy-at-msa.microscopy.com} ; Sat, 2 Dec 1995 14:04:17 -0600 } Subject: dusrt part needed also } To: microscopy-at-Sparc5.Microscopy.Com } From: "Robert Schmitz, Biology" {rschmitz-at-macsrv1.uwsp.edu} } Date: Sat, 2 Dec 95 14:08:42 +2000 } Message-ID: {951202.140842.13419-at-macsrv1.uwsp.edu} } Content-Type: text } } } } Subject: Durst enlarger parts } } } } Hello on the net: } } } } I am desperately trying to find someone who has a Durst Laborator } } S-45 EM enlarger that they no longer need. We are particular } } interested in finding the NEGA-138 carriers with glasses. Also the } } upper portion containing bellows and focusing devices. Any help would } } be very much appreciated. We have two enlargers and need to keep } } them in operational order. Thanks Cathy Kelloes } } Cathy isn't the only only individual looking for Durst S-45 parts. I am } looking } for NEGA-138 carrier for 4x5 negatives and a Carlwen negative carrier (in } particular, the negative carrier for 35mm film). I also would like to find an } extra set of Latico 240 and 200 lens, as on of my colleagues claims ours has } a } hot spot that shows up in enlargements 3x or greater. Please reply to Cathy } first but then if you can help me please contact me. Thanks!! } } rschmitz-at-macsrv1.uwsp.edu } Bob Schmitz, Biology, } Univ. WI Stevens Pt. } Stevens Point, Wisconsin 54481 } ph 715-346-2420 } }
} X-UIDL: 28f9cf215b1570116f02621cd0a5625d } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id } TAA00475 for dist-Microscopy; Sat, 2 Dec 1995 19:41:18 -0600 } Received: from hermes.bcm.tmc.edu (HERMES.BCM.TMC.EDU [128.249.2.129]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id TAA00472 for } {Microscopy-at-sparc5.microscopy.com} ; Sat, 2 Dec 1995 19:41:17 -0600 } Received: from bcm.tmc.edu (aburns-at-bcm.tmc.edu [128.249.2.1]) by } hermes.bcm.tmc.edu (8.7/8.6.11) with SMTP id TAA05389 for } {Microscopy-at-sparc5.microscopy.com} ; Sat, 2 Dec 1995 19:40:21 -0600 (CST) } Received: (from aburns-at-localhost) by bcm.tmc.edu (8.6.11/8.6.6) id TAA04633; } Sat, 2 Dec 1995 19:40:16 -0600 } Date: Sat, 2 Dec 1995 19:40:15 -0600 (CST) } From: "Alan R. Burns" {aburns-at-bcm.tmc.edu} } X-Sender: aburns-at-watson } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: RE: TEM of Sorghum leaves (fwd) } Message-ID: {Pine.SOL.3.91.951202193827.4291C-100000-at-watson} } MIME-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } } } } ---------- Forwarded message ---------- } Date: Sat, 2 Dec 1995 19:28:06 -0600 (CST) } From: Alan R. Burns {aburns-at-bcm.tmc.edu} } To: Phill {aarwharn-at-reading.ac.uk} } Subject: RE: TEM of Sorghum leaves } } Phill: } } Many years ago I used an ultra-low viscosity medium for rapid embedding } of plant cells. This resin prevented plasmolysis and the tissue } did not separate from the plastic. We published our findings and } the recipe for this plastic (available from Polysciences) in the } following article: } } Oliveira et al., 1983. J.Microscopy 132:195-202. } } Give it a try, it might just do the trick. } } }
} X-UIDL: 9d73692d7860aab95c5b4f95717eed4d } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id } UAA00505 for dist-Microscopy; Sat, 2 Dec 1995 20:06:02 -0600 } Received: from mail-relay1 (mail-relay1.cis.yale.edu [130.132.21.199]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id UAA00502 for } {Microscopy-at-sparc5.microscopy.com} ; Sat, 2 Dec 1995 20:06:01 -0600 } Received: from quickmail.yale.edu ([130.132.144.201]) by mail-relay1 with SMTP } id AA05608 } (5.67a/IDA-1.5 for {Microscopy-at-MSA.Microscopy.Com} ); Sat, 2 Dec 1995 } 21:03:51 -0500 } Message-Id: {n1394184190.65950-at-QuickMail.Yale.edu} } Date: 2 Dec 1995 21:04:25 -0500 } From: "Paul Webster" {Paul.Webster-at-quickmail.yale.edu} } Subject: Re: TEM of Sorghum leaves } To: "Microscopy BBS" {Microscopy-at-Sparc5.Microscopy.Com} } X-Mailer: Mail*Link SMTP-QM 3.0.2 } Content-Type: text } } Subject: Time: 8:46 PM } OFFICE MEMO Re} TEM of Sorghum leaves Date: 12/2/95 } } In message {Pine.SOL.3.91.951129162920.2689F-100000-at-suma3.reading.ac.uk} Phill } writes: } } } } Hello, } } I have recently started doing TEM on the infection of sorghum leaves by } } the fungus Colletotrichum graminicola. However, I have not had very much } } success as of yet because I have had a lot of trouble fixing and } } sectioning the leaf tissue. My main problems at the moment are poor } } fixation and the resin splitting away from the leaf along the cuticle } } when I am sectioning. } Phil, } } } The aldehydes normally used for EM fixation have been selected because they } work } well for mammalian tissue. They fix well because the material is full of } primary amines which the aldehydes can crosslink. Plant tissue is very } different and is therefore not easy to fix with these chemicals. Some } suggestions which I have heard to solve the problem of using these chemicals on } plants include adding extra primary amines to the material just prior to the } fixation (this is the theoretical basis of Nakane fix, which has lysine added } to } the fixative solution, and to the fixative described by Luther & Block). } } Another good idea is to cool the plants down by leaving then in a fridge or } cold } room for a few hours and then fixing them in cold fixative This method I } collected from a plant biologist, so I know it works. The logic behind the } method can be easily demonstrated by trying to fix 2% gelatin at room temp. It } will not fix because it is a liquid, but if it is cooled, the gel formed can be } crosslinked by aldehyde. The fixed gelatin can be easily cut up and embedded, } so should the leaves. } } Another alternative is to rapidly freeze the material by high pressure } freezing, } Muller et al (J. Microsc.) did this with apple leaves and achieved remarkable } morphology in the fully hydrated sections they cut afterwards. Although it is } technically demanding to freeze the material, the leaves need not be } cryosectioned, but could instead be embedded in resin by freeze substitution - } a } process that only takes a day or two at most using simple materials. } } Whatever method you choose, we wish you good luck. } } Paul Webster } Center for Cell Imaging } Yale School of Medicine } http://info.med.yale.edu/cellimg } }
} X-UIDL: 808c89fcc94947d5a9eef06f9ae8f364 } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id } XAA00631 for dist-Microscopy; Sat, 2 Dec 1995 23:12:18 -0600 } Received: from apollo.roundlake.baxter.com (apollo.roundlake.baxter.com } [159.198.1.9]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id XAA00628 } for {Microscopy-at-MSA.Microscopy.Com} ; Sat, 2 Dec 1995 23:12:17 -0600 } Received: from msmail_smtp.roundlake.baxter.com by } apollo.roundlake.baxter.com via Pony Express SMTP with TCP } (v8.1.1-dmr001); Sat, 2 Dec 95 23:02:25 CST } Received: by msmail_smtp.roundlake.baxter.com with Microsoft Mail id } {30C14B8D-at-msmail_smtp.roundlake.baxter.com} ; Sat, } 02 Dec 95 23:02:37 PST } From: "Neuberger, Damian" {neuberd-at-engrnd.roundlake.baxter.com} } To: "'Microscopy '" {Microscopy-at-Sparc5.Microscopy.Com} } Subject: TEM of Sorghum Leaves } Date: Sat, 02 Dec 95 23:10:00 PST } Message-ID: {30C14B8D-at-msmail_smtp.roundlake.baxter.com} } Encoding: 30 TEXT } X-Mailer: Microsoft Mail V3.0 } Content-Type: text } } } I don't have Phil's original message or address but would like to add a few } comments to the already good ones submited. } } You should not be having a problem fixing Sorghum using typical plant } fixation protocols, but without the specific details I can't comment } further. However, a protocol for fixing Pinus resinosa seedlings, which } were very difficult to fix and embed properly, utilized a 6% v/v } glutaraldehyde and 6% paraformaldehyde v/v in 0.08 M cacodylate buffer at pH } 7.2 for 2 hr. Following further dissection (I was looking at the phloem } sieve elements), the tissue was additionally fixed for 10 hr at RT with a } change of fresh fixative every hour. During this fixation step, the vials } were on a rotator for continuous mixing. The tissue was washed in buffer } and postfixed in cacodylate-buffered 2% OsO4 overnight in a refrigerator } The reason for the changes is the presence of phenolic compounds that were } reacting with the aldehydes (according to a biochemist). As for the poor } infiltration of embedding medium, I used Spurr's low viscosity resin and } diamond knives. } } If want to discuss this further and/or get references, we can do that } off-line. Hope this helps. } } Damian Neuberger, Ph.D. } Research Scientist } Baxter Healthcare, Corp. MS WG3-2S } P.O. Box 490 } Round Lake, IL 60073 } Tel: 708.270.5888 } Fax: 708.270.5897 } EMail: neuberd-at-baxter.com } }
} X-UIDL: c88639b22b35c7bd092a5248e3c00497 } Received: from strauss.udel.edu (strauss.udel.edu [128.175.13.74]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id IAA00924 for } {listserver-at-sparc5.microscopy.com} ; Sun, 3 Dec 1995 08:09:35 -0600 } Received: (from xianying-at-localhost) by strauss.udel.edu (8.6.12/8.6.12) id } JAA16551; Sun, 3 Dec 1995 09:08:34 -0500 } Date: Sun, 3 Dec 1995 09:08:34 -0500 (EST) } From: Xianying Burany {xianying-at-UDel.Edu} } To: listserver-at-sparc5.microscopy.com } Subject: Kevex & Tracor Northern } Message-ID: {Pine.SOL.3.91.951203085038.14180B-100000-at-strauss.udel.edu} } MIME-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } } I am looking for the email addresses of Kevex and Tracor Northern. I need } help for our two EDSs. Please reply directly to me at } } xianying-at-udel.edu } } I greatly appreciate your help. } } Sandy Burany, PhD } Dept. of Physics and Astronomy } University of Delaware } Newark, DE 19716 } (302) 831-3515 } Fax: (302) 831-1637 } }
} X-UIDL: 4527741dda7cffcd01422f0c149d3b9a } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) id } NAA01044 for dist-Microscopy; Sun, 3 Dec 1995 13:01:15 -0600 } Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id NAA01041 for } {Microscopy-at-sparc5.microscopy.com} ; Sun, 3 Dec 1995 13:00:50 -0600 } Date: Sat, 02 Dec 1995 13:12:40 -0500 (CDT) } Date-warning: Date header was inserted by uthscsa.edu } From: "Nancy K. R. Smith" {SMITHN-at-UTHSCSA.EDU} } Subject: Al signal in EDS } X-Sender: SMITHN-at-arweN.UTHSCSA.EDU } To: microscopy-at-aaem.amc.anl.gov } Message-id: {01HYC02TNILU00AFQC-at-uthscsa.edu} } X-Envelope-to: microscopy-at-aaem.amc.anl.gov } MIME-version: 1.0 } X-Mailer: Windows Eudora Light Version 1.5.2 } Content-transfer-encoding: 7BIT } Content-Type: text/plain; charset="us-ascii" } } } Date: Fri, 01 Dec 1995 19:19:05 } } To: jeff_jones-at-mindlink.bc.ca } } From: "Nancy K. R. Smith" {SMITHN-at-UTHSCSA.EDU} } } Subject: Al signal in EDS } } } } } } Tania Jones, c/o Jeff Jones: } } The extraneous Al signal comes from the x-ray detector housing. X-rays } and backscattered electrons strike the housing and generate Al x-rays. Some } of those originating from just in front of the Be window are able to have } line-of-sight entry into the window and be detected. } } } .............................................................................. } .Post note: My email addresses are as follows: } work nkrsmith-at-juno.com } home smithn-at-uthscsa.edu } BEST STRATEGY: address mail to smithn-at-uthscsa.edu and cc it to } nkrsmith-at-juno.com. (Ignore the fact that the work & home addresses seem } backwards.) } .............................................................................. } .Nancy K.Rodman Smith, Ph.D. } .Dept. Cellular & Structural Biology } .University of Texas Health Science Center at San Antonio } .7703 Floyd Curl Drive } .San Antonio TX 78284-7762 } .210-567-3861 Fax:210-567-3803 } .Email:SMITHN-at-UTHSCSA.EDU; NKRSMITH-at-JUNO.COM; NKRS-at-JUNO.COM } .............................................................................. } "God put me on earth to accomplish a certain number of things. Right now I } am so far behind that I will never die." (Calvin, from Calvin & Hobbes } cartoon) } **************************************************************************** } ***** } }
} Date: Fri, 01 Dec 1995 19:19:05 } To: jeff_jones-at-mindlink.bc.ca } From: "Nancy K. R. Smith" {SMITHN-at-UTHSCSA.EDU} } Subject: Al signal in EDS } } } Tania Jones, c/o Jeff Jones: } The extraneous Al signal comes from the x-ray detector housing. X-rays and backscattered electrons strike the housing and generate Al x-rays. Some of those originating from just in front of the Be window are able to have line-of-sight entry into the window and be detected. } .............................................................................. .Post note: My email addresses are as follows: work nkrsmith-at-juno.com home smithn-at-uthscsa.edu BEST STRATEGY: address mail to smithn-at-uthscsa.edu and cc it to nkrsmith-at-juno.com. (Ignore the fact that the work & home addresses seem backwards.) .............................................................................. .Nancy K.Rodman Smith, Ph.D. .Dept. Cellular & Structural Biology .University of Texas Health Science Center at San Antonio .7703 Floyd Curl Drive .San Antonio TX 78284-7762 .210-567-3861 Fax:210-567-3803 .Email:SMITHN-at-UTHSCSA.EDU; NKRSMITH-at-JUNO.COM; NKRS-at-JUNO.COM .............................................................................. "God put me on earth to accomplish a certain number of things. Right now I am so far behind that I will never die." (Calvin, from Calvin & Hobbes cartoon) **************************************************************************** *****
When looking into source of the extraneous Al signal you should also investigate the composition of your collimator to your EDS detector. You may have electrons hitting the collimator creating x-rays which are then detected by the SiLi system. This should be checked in addition to the other components within the EM Chamber.
BTW, I presumed that you have a collimator, if not that could also be part of the problem.
Also if you have a retractable EDS detector is the Al signal a function of the detector position?
We regularly use LR White for immunocytochemistry of plant and microbial material. Our schedule is as follows. Fix briefll ( the smaller the sample the shorter the fixation- but typically 1H) Fixative is 2%paraformaldehyde + 0.5%glutaraldehyde in 50mMPIPES ph &.2-- you may have to adjust this to your specimen) Dehydrate Ihour in each of 30, 50 70 and 80% EtOH at +4;-15;;-25 and again -25oC. Leave for 1H in 50:50 80% EtoH;LR White. Leave overnight in the fresh 50:50 mixture, one hour in fresh 50:50 mixture. Embed in fresh resin (Do all resin changes in an inert atmosphere) Polymerize at -25oC for 24-48 hours. then place capsules in a window to get what passes for sunlight in a Cambridge winter. Immuno- schedule is 3 x 5min in a blocking buffer; 4-16 hours in primary antibody, wash x3; 90mins in a biotinylates secondary Ab; washx3; 90 mins in streptavidin label (FITC,Texas Red colloidal gold etc.; wash x3 Ready to go. If you want detailds of various solutions e-mAil me direct
With Seasons Greetings
Patrick Echlin Director Multi-Imaging Centre Schhol of Biological Sciences University of Cambridge United Kingdom
On Fri, 1 Dec 1995, Christine A Brantner wrote:
} Hello } } I am a graduate student at the Univeraity of Wisconsin-Milwaukee and I have } a problem with a technique. } } I embed bacteria in LR White. I did ultrastructural work and stained with } UA and Pb and had no major problem with holes in the resin. The next } project is to immunostain a protein in the membrane. I can use the same } blocks as before and after immunostaining, there are multiple holes in most } cells. I put the sections on nickel grids. NaCl for 1 min. New BSA 3% } filtered for 1 min. Primary antibody for 1 hour. BSA washes. Secondary } antibody IgG for 1 hour. BSA washes. NaCl wash. Water washes. The I } stain with UA for 1 hour as per usual. } } Help.....I'd like to get rid of the holes. I think that it is happening } during this procedure. The new BSA has not eliminated them as was suggested } that it might. } } Thank You } } Chris Brantner } } cab-at-csd.uwm.edu } University of Wisconsin-Milwaukee }
We have a grad student looking for an Image Analysis program to run on a PC. He is looking at polymer images on an optical microscope and wants to measure particle size, distribution and fibre length.
In addition of the other good ideas, try to use 0.18 M PIPES buffer at pH 6.8 vith 0.5 to 1.0% Caffeine, in BOTH fixative (ald. & osmium) Prepare solutions immediately before use. Extra long fixation times! All the best,
} You may "paint" your collimator inside and outside with pure } colloidal graphite (carbon paint) to minimize extraneous Al signal. } } Laszlo J. Veto
Make sure it's pure graphite! I've found carbon paints sold for mounting specimens for EDX that are contaminated with phosphorus . Phil Oshel
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
35mm film: 37TB frame to fit L-35 base, if you don't have the base, you have to order the L-35 base with opening for 35mm film. The frame has focusing target and both are black anodized. At Carlwen I talked to a Mr. Locken.
As I said, I do not know if either of the 3 companies are still in business or if they have moved. My P.O.s go back almost 10 years. It's certainly worth a try. Hope this helps.
Peace,
Phil Rutledge, Director Center for Electron Microscopy UMBC Dept. of Biology Catonsvill, MD 21228
A few thoughts on Melanie Behrens' posting regarding the aluminum peak in the EDS spectrum of the iron oxide:
First, I would agree that the problem is in the specimen and not an artifact from the microscope/detector since your colleague can obtain the same result using a different instrument. You might also try analyzing a sample that you are absolutely certain contains no aluminum using the same instrument settings (accelerating voltage, count rate, etc.) and see if the aluminum peak appears.
You also commented, "The sample was ground up but not prepared in epoxy and polished or coated, since we are not looking for a strictly quantitative result and want something of a bulk analysis as fast as possible." Are you running this truly quantitative (comparing it against a known standard) or are you using a "standardless miracle" routine, as the DTSA folks call it? This could cause you a lot of headaches.
Since you ground up the sample, and are after a bulk analysis, wouldn't it make more sense to analyze it via XRF?
Jim Stets Air Products and Chemicals, Inc. Allentown, PA stetsjr-at-apci.com
Greetings: Although I appreciate that fact that information from a plant biologist can be considered as gospel truth, I would just like to point out that the effects of cold treatment (chilling) can be considerable, from both a metabolic and morphological point of view. I would suggest that the first hour or so of the fixation should be carried out at room temperature, followed by an equal length of time (or longer) at 4 C. The initial fix at room temperature will act to preserve the tissue without introducing any structural variation due to chilling. I have a protocol that I can provide, that works well with dicot leaves. If interested drop me a note and I'll send it off. Damian makes reference to the use of both para- and glutaraldehyde to fix difficult tissue. I also use both in standard chemical fixations to enhance the likelyhood of rapid and adequate fixation. I use a lower percentage of PA, but it will vary according to the tissue type and the desired level of fixation. Best of luck with the fixation.
Dwight Beebe Institut de recherche en biologie vegetale beebed-at-ere.umontreal.ca Dept. de sciences biologiques Voice:514-872-4563 Universite de Montreal FAX:514-872-8496 4101, rue Sherbrooke est Montreal, PQ H1X 2B2 Canada
Dear microscopists, I need some information about fixation and contrast improvment of ovocyts and spermas for ESEM. I have no experiences with such of type of specimens. I ask you to give me some advices about preparing and technique. Thank You.
=========================================================== Jiri Spinka Faculty of Electrical Engineering and Computer Science Department of Electrotechnology Technical University of Brno EEEEEE TTTTTT Antoninska 1, B R N O EE TT Czech Republic EEEE TT Tel. 42-5-753741, Fax. 42-5-41211135 EE TT e-mail: spinka-at-uete.fee.vutbr.cz (Internet) EEEEEE(hi) TT ===========================================================
I am sending this response to the listserver so that each of those who wanted further details about the problem could get a more detailed description.
The detector is a Noran thin window light element detector with 134 eV resolution, with a Voyager II workstation. The detector has not been serviced for about a year, so cleanliness or ice buildup is possible, but the problem does not occur with all iron oxides or mixtures of metals with light elements, only with specific samples. A different instrument with a Kevex detector at another of our company's locations has also experienced this problem with the same samples, so I believe it is a sample problem, not a detector problem. The background shape appeared to be normal.
The sample may be a mixture of oxides, but the stoichiometry is so far off that even FeO would not account for it, and the XRD (if I remember correctly, since it has been awhile since I last saw this phenomenom) did not report FeO exclusively. I will check on this tomorrow at work. I will also check what other elements were present, but I believe the EDS gave the result as over 90 wt% iron, with perhaps 5 wt% oxygen, if that. The sample was ground up but not prepared in epoxy and polished or coated, since we are not looking for a strictly quantitative result and want something of a bulk analysis as fast as possible. I think it is natural iron oxide, since we did not synthesize it in the lab - it was found in an unwanted location. It did not appear to be iron metal either by XRD or BSE image.
Hope this answers some of your questions and encourages more discussion. Thanks to all who have already replied to my posting.
} i frequently have Al detected when doing EDXA on plain carbon } steel samples, even though i mount the samples using a carbon- } based adhesive. However, i do not detect it all the time. any } suggestions as to why this is occurring? } } thanks, } } tania jones Is the mounting stub made of Al and the sample 'relatively' thin? Recall the "volume of interaction" is typically greater than what you are viewing at 1000x...
Best regards,
Mark T. Biedrzycki Computer Networking Laboratory Materials Science and for Microscopy Education (CNLME) Engineering Department at the University of Arizona, Tucson
"When phenolic-containing plant cells are fixed with glutaraldehyde followed by OsO4, phenolics leach from the vacuoles into the cytoplasm where they subsequently react with OsO4. The result is a dense, osmiophilic cytoplasm, the details of which are obscured. This leaching can be prevented by fixation with glutaraldehyde containing 0.1-1.0% caffeine and then a rinse with a buffer containing caffeine."
Ref.: Hayat, M. A. (1986). "Basic Techniques for Transmission Electron Microscopy" Academic Press. Inc., Orlando, Florida. ISBN 0-12-333925-1 Hope it helps, regards,
Hello all, I would be grateful if anyone could point me in the direction of companies that manufacture or deal in tissue processors, for histological (paraffin embedding) use. I have turned up nothing through our sources here; Fischer discontinued their line. Thank you in advance, Pat Reynolds ___________________________ Patrick D. Reynolds Biology Department Hamilton College 198 College Hill Rd. Clinton, N.Y. 13323 Voice: (315) 859-4723 Fax: (315) 859-4807
Anyone out there have a good technique for obtaining fracture surfaces of fine (10-20um dia) fibers that remain flexible even at LN2 temps? Trying to get SEM images of fracture sections without smearing associated with microtomy or other cutting. Thanks
Anyone out there have a good technique for obtaining fracture surfaces of fine (10-20um dia) fibers that remain flexible even at LN2 temps? Trying to get SEM images of fracture sections without smearing associated with microtomy or other cutting. Thanks
We handle all types of repairs, modifications and custom design work for microscopy. We also do a lot of Imaging and aquisitions from the microscope. Please call us if you have any questions.
Yes, I am using a standardless PROZA routine and yes, I realize this is not the ideal way of performing quantitative analysis. However, I have gotten acceptable although not perfect results with other samples containing iron oxides. I think, however, that you may be right about the sample effects of grain size and orientation possibly being the reason for the problem. Thanks, Melanie Behrens Senior Chemist Texaco, Inc
You wrote: One thought to check for instrumental vs. sample effects in your anomalous Fe-oxide is to look not only at the Fe/O ratio, but also the Fe-K/Fe-L ratio (assuming that you are analyzing Fe by measuring the K-alpha line). Because of my WDS background I don't have a good opinion how well crushed samples work, but consider that with over a factor of 10 difference in energy, Fe and O information does not come from strictly the same part of the sample. Grain size or even grain orientation might make a big difference in the relative sizes of the Fe and O peaks. I studied this effect once (although unfortunately I don't have the data at hand any more) with Si, Ca, and Fe (much more similar to each other in energy than Fe and O), and concluded that it was hardly possible to distinguish Ca3Fe2Si3 and CaFeSi2 unless I was working with a surface of known orientation!
A further question--when you say over 90% Fe, is that relative to a standard, or are you using a "standardless" precedure that normalizes to 100% (another thing that I distrust with my WDS bias).
Best wishes, Alfred --- Alfred Kracher akracher-at-iastate.edu http://www.public.iastate.edu/~akracher/
} We have a grad student looking for an Image Analysis program to run } on a PC. He is looking at polymer images on an optical microscope and wants } to measure particle size, distribution and fibre length. } } Tom Bryner } }
Has anyone got a suggestion for an alternative transition fluid to replace the now banned and unavailable halocarbons which have been used in critical point drying using either ethanol of acetone. I'm not sure I want to use amyl acetate as its a bit toxic and remains in the seals of the critical point dryer. For Cambridge watchers, it's two degrees celcius and snowing
Patrick Echlin Multi-Imaging Centre School of Biological Sciences University of Cambridge.
Caffeine takes care of the poly=phrnols which abound in many plant tissues.
Patrick EchlinOn Mon, 4 Dec 1995, Jay Jerome wrote:
} } Lazlo- } } Why is caffeine added to the buffer? } } } Jay Jerome } ************************************************************** } * aka: W. Gray Jerome * } * Dept. of Pathology * } * Bowman Gray School of Medicine of Wake Forest University * } * Medical Center Blvd * } * Winston-Salem, NC 27157-1092 * } * 910-716-4972 * } * jjerome-at-bgsm.edu * } ************************************************************** } } On 4 Dec 1995 VETO-at-BCRSSU.AGR.CA wrote: } } } Phill: } } } } In addition of the other good ideas, try to use 0.18 M PIPES buffer at } } pH 6.8 vith 0.5 to 1.0% Caffeine, in BOTH fixative (ald. & osmium) } } Prepare solutions immediately before use. Extra long fixation times! } } All the best, } } } } Laszlo J. Veto } } }
Message-Id: {199512061509.KAA14889-at-post-ofc02.srv.cis.pitt.edu} To: "microscopy-at-aaem.amc.anl.gov" {microscopy-at-aaem.amc.anl.gov}
Does anyone know of a PPM to TIF converter (freeware, shareware or even commercial)?
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
please add me to your list: Ronald F. Mervims, Ph.D. (Ron Mervis-at-aol.com) President NeuroMetrix Rearch, Inc. 2109 West Fifth Ave., suite B Columbus, Ohio 43212
I am trying to find information on a cement microscopy conference to be held in Houston (?) this February (?). Any information would be appreciated. Thanks...
Larry Sutter Michigan Technological University Dept. of Civil and Environmental Engineering 1400 Townsend Dr. Houghton, MI 49931
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
We have in our laboratory a client who has a soft contact lens (polyhydroxyethyl methacrylate) that has been exposed to a dye, of the type that gives soft lenses different colors. The question is one of determining to what degree the dye is chemically bonded to the polymer vs. being entrapped in the polymer's porosity.
The less than 0.1 Wt. % of dye loading would seem to me to preclude any possible use of XPS. While SIMS is a lot more sensitive, I would not have thought one could get useful information of this type at these levels.
Wondered if anyone out there has ever been confronted with this kind of a problem and would offer any suggestions.
Thanks in advance.
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: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
} Has anyone got a suggestion for an alternative transition fluid to } replace the now banned and unavailable halocarbons which have been used } in critical point drying using either ethanol of acetone. I'm not sure I } want to use amyl acetate as its a bit toxic and remains in the seals of } the critical point dryer. } For Cambridge watchers, it's two degrees celcius and snowing } } Patrick Echlin } Multi-Imaging Centre } School of Biological Sciences } University of Cambridge.
We use acetone or in some cases ethanol straight through to the CO2 and for most purposes it works just as well. We increase the times a bit and make sure that the solvents are as waterfree as possible. If it is very critical you will need a molecular sieve to dry the solvent and also a water trap for the CO2.
We also tried dimethoxypropane some years ago. It works both for dehydration (it forms acetone and methanol when mixed with water) and subsequently as transition fluid. As I recall the result was about the same as with the normal procedure (dehydration with ethanol and Freon as transition fluid).
Hope this helps.
Uppsala is not any better, it's been down to -10 celcius the last couple of days and generally unpleasant.
If you get direct posts, would you please forward them to the listserver. I too, am interested in a substitute, and have not been very happy with the quality of the CPD material since we can't realistically use Freon TF. I am under the impression that even freshly opened alcohol, within a few seconds absorbs water, but it may be even more complex than that, such as miscibilities of alcohol and CO2. We have had numerous suggestions such as drying the CO2 tank out before refilling at the supplier, but I am afraid that our supplier is not that cooperative. To go the the next better grade of CO2 the price per tank goes from $15.00 to $130.00, and the specified composition with regard to water improves only marginally (I believe from 0.2% water to less than 0.1% water). A rather pricey experiment. We have a drier in the CO2 line and condense CO2 gas into the chamber, rather than use a siphon tube but these do little to improve the quality. Peldri II does work slightly better than CPD but does seem somewhat material dependent, and is relatively more expensive. Lastly, it should be mentioned that we do a large number of samples in a Polaron Jumbo CPD, so we do go through a fair amount of reagents. I have been reluctant to use anything thus far besides alcohol both from the point of view of flammability and toxicity. During the winter months, with low humidity, on release of the CO2 there is a lot of static discharges. Knock on wood, no ignitions yet, but I am concerned. Any responses that appear or that you pass along will be appreciated. later dlb
David Bentley Imaging Facility, Div. of Biotechnology, ARL The University of Arizona Tucson, Az 85721
for the Hitachi Office supporting Acoustic Microscopes ?
I need a manual for model AT-5000
Thanks Much !
Regards,
Ed Monberg, GM em-at-mediacity.com LMDC (Laser Motion Development Company) 3101 Whipple Road Union City, CA 94587-1216 510-429-1060 voice 510-429-1065 fax
Several months ago I asked a question about solder analysis via EDS. I promised to post a summary of the replies and had not done so. Here are the replies I received. I enclosed the majority of the replies. You will have to judge the techniques yourself, since we have not progressed any further in our quest, due to time and/or budget constraints.
I apologize for not doing this sooner.
If you get any good results please let me know.
John Giles
Here's the original post followed by the replies: ------------------------------------------------------------------------------
Hello,
I am interested on the community input on performing in-situ analysis of solder to determine composition. We have tried to do this in the past and have not had acceptable results. We used both multiple areas and multiple points and averaged the data.
I realize that this is probably an impossible task with EDS, but would like to confirm my opinion and obtain input as to any other techniques (micro probe XRF, etc.) which may provide a non-destructive method for in-situ solder analysis.
Thanks,
John Giles jegiles-at-space.honeywell.com ---------------------------------------------------------------------------
The beast raises its ugly head again.
Yes, you can get quant info this way. I have done it, but it is not very accurate. If you have a reference sample of a known solder you can use image processing to get a good idea of the solder composition you are examining. This is purely subjective of course, which is why you need a reference sample. You need to do a histogram analysis of the unkown and reference samples under the same conditions. The method you use to get the numbers to look correct on the reference is repeated on the unkown sample. This will usually get you ballpark type numbers, but again it is purely subjective. We can discuss this method further if you wish.
A far better way is to use an XRF thickness machine. There are standards available for the common binary solders. This method is much faster. We have gotten bars of trinary solders chemicaly analyzed and plan to set up calibration files for these solders. So far no one has had the time to do this.
What is your objective? Are you trying to verify the type of solder used or
are you trying to profile the change in composition under different conditions?
Al ----------
Al We have been getting a lot of conflicting data about analyzing solder on pwb's. Some people are claiming that they can get quant info by a combination of image analysis and eds. Know anything about this?
Ron -------------------------------------------------------------------
John, It seems to work best when I can polish the sample and look at an area at very low magnification. Low magnification being 100X in the SEM. For instance, making a polished section to check the composition in the solder pot for solderability testing. In general, the lower the magnification, the better the overall results. Looking at points hasn't worked.
A fellow metallurgist at GE Cincinnati says that you have to take photographs of many different areas, drop a ruler and get grain size and/or volume fraction of the two phases, and then calculate the composition. A backscatter detector would probably work the best to highlight the two phases. Old fashioned image analysis. Some of the new EDS systems have these capabilities-ours doesn't.
You are certainly correct with regard to homogeneity, but enough area samplings should be somewhat representative. We have a Link LZ5 detector and a Dapple System analyzer/software. Given consistent operating conditions, sample condition, good reference standards, etc, you should be able to get good results.
We have a good relationship with our supplier, Dapple, and have run quant studies for them. Though I had extreme difficulty with Kevex in the past, I believe those individuals have been replaced. Have you tried their tech support?
I will give you Bill Stewart's (Dapple System) telephone number. If Kevex cannot or has not helped you, then try calling him. He can tell you what limitations you can expect from EDS in general and if your Kevex system is insufficient for the task. If you do call Bill, tell him I suggested you contact him. I have always found him to be very helpful.
Bill Stewart Dapple Systems 408/733-3283
Please send me an update. Good Luck.
Alan Stone ASTON Metallurgical Services ------------------------------------------------------------------------
John Giles,
I am forwarding you some information I rec'd from a faculty member here in the Metallurgy department at Michigan Tech. Hope this is some help to you.
Owen P. Mills Dept. of Metallurgical & Materials Engineering Michigan Technological University Houghton, MI 49931 906-487-2002 opmills-at-mtu.edu
---------- Forwarded message ----------
Owen:
I haven't done any work, but have maintained a passing interest in soldering as related to wetting during joint formation. Aside from the standard SEM/EDS references, the following book (though not directly addressing the problem) might have some secondary references related to analysis of solders:
Solder Mechanics-A State of the Art Assessment D. R. Frear, W. B. Jones, and K. R. Kinsman TMS, Warrendale, PA
Authors of the various articles will also be good sources for information. I know that several of them are from Sandia Albuquerque which has had a large program on soldering for several years.
Cal Calvin L. White Department of Metallurgical and Materials Engineering Michigan Technological University Houghton, MI 49931 phone (906) 487-2036 fax (906) 487-2934 email cwhite-at-mtu.edu ---------------------------------------------------------------------
Hello John, Here at Princeton we have a 6 spectrometer (5 WDS/EDS) EPMA system that we use for a wide range of nondestructive analyses. ALthough I'm unfamiliar with your samples I see no reason your problem couldn't be solved by wavelength spectroscopy. If you are interested you could send me a specimen and I could produce some results for you. Just let me know. Ed Vicenzi
I have had a lot of problems determining the composition of solder with EDS. If life is good, and we process escape peaks, remove the background and quantify, the results will be very close. It is impossible if it is high temperature solder which may include 1-2% silver, whose peaks are severely overlapped with lead and tin. Since we usually just want to know the melting point of the solder, and the samples can be destroyed, we are now using differential scanning calorimetry (DSC) to determine the solder melting point. This can be correlated to the composition of standard tin-lead solder. The sample size is relatively small and the equipment is comparatively inexpensive. Most service labs have it also.
Not quite what you asked, Sharon -------------------------------------------------------------------------
John, Having been frustrated by solder I thought I would pass on my "wisdom." The problem with trying to analyze solder by any microbeam technique is the nature of the material as it solidifies. I assume you are dealing with a tin-lead system on the lead rich side of the eutectic composition. Because solder solidifies through the same "pasty" range (liquidus-solidus region above the eutectic temperature and below the liquidus temperature) it went through on fusion the composition will vary throughout the mass beginning where the first nucleation occurred and ending with the material that solidified last. If you follow the solidification process on the Pb-Sn equilibrium diagram you will get an idea of the potential problems brought on by compositional variations. Also, when solder solidifies the size of the individual grains will vary due to localized compositional variations, and more importantly the rate of change of temperature during the solidification process. From previous experience in attempting to do semi-quantitative analysis of solders using an SEM/EDS approach we soon discovered it is not practical to do it in situ. Our procedure was to carve away a good size chunk of solder, place it between two steel plates then flatten and thin it out with a healthy "calibrated" hammer blow. The flattening reduced the compositional variation due to depth by spreading it out and making it more accessible to excitation. Collecting a number of spectra which covered most of the surface area which would be summed together and used for "quantification" gave reasonable results when compared with known materials handled in the same way. Trying to use a beam in spot mode, whether in an SEM or EPMA, would not work because of the uncertainty of which phase in the solder the beam is exciting. XRF holds a little better hope, particularly if you can use the higher energy lines, i.e., Sn -K and Pb-L for analysis and have an EDXRF system. However, what do you use for standards? and how do you account for geometry affects? Wavelength methods would be very difficult without precise geometry relationships.I think you get the point that I don't believe in situ is the way to go!
I hope this input has been of some value to you, and good luck! I would be interested in hearing how you make out with this search, could you post a summary of responses.
Bob Craig OSRAM SYLVANIA INC. Lighting Research Center Beverly, MA 01915 craig-at-hq.sylvania.com ---------------------------------------------------------------------------
John, We're looking at approx 0.5 mm dots of solder mounted on aluminum stubs. We haven't needed to check solder on circuit boards, but that ought to be possible with appropriate grounding. The WDS has been used to look at oxygen and some trace elements. We use ZAF quantitation with EDS to look at ratios of lead and tin. We haven't done any absolute quantitation with standards, but that ought to be possible too. Would you like to send a specimen here and see what our spectra look like? Mike ---------------- } Thanks for your reply. The specific application I am looking at is analyzing } solder joints on a PC board. One of our production engineers would like to use } this as an inspection technique for solder composition as opposed to our } typical analysis of bulk samples from the solder pot prior to soldering. I do } not have a WDS on our system, but would be interested in the results you get on } a known solder standard. } } John
John, At MCNC, we've been analyzing solder using a combination of EDS and WDS. Those techniques are nearer to bulk composition analysis than the surface-oriented techniques are. Maybe I didn't understand exactly what you need to analyze. If you want to send more details, I could pass on your questions to our group that works on solder. Perhaps they can help. Mike Lamvik
John, We perform analysis of solders quite frequently here at Sandia National Laboratories. I do not believe that the average or bulk composition of the solder can be determined by scanning. What we have done with good success is to determine the volume fractions of the phases present in the solder and then determine the composition of each of the phases. At this point it is a fairly straight forward exercise to calculate the bulk solder composition. Please contact me if you have any questions.
Joe Michael jrmicha-at-sandia.gov -----------------------------------------------------------------------------
John,
We have performed the analyis on our AMRAY 1645 which is equipped with a large specimen stage and a Noran x-ray system. Our particular application was to determine the composition of solder as it flowed down a copper conductor on a printed circuit board. The solders we were interested in were Pb-Sn and I think it was 60-40. Anyway, we used the image analysis on the Noran to determine the
area fraction of the two phases on the surface of unpolished solder lines. Then we very carefully measured the composition of the two phases. We also made the assumption that the volume fraction of the two phases was equivalent to the
area fraction that we measured. At this point you need to write a simple equation that allows you to calculate the bulk composition of the solder from the volume fractions and the compositions of the phases. You will also need to
know or assume a density for the phases. We published some results from this technique in Acta Met vol, 43, pp 299-305, 1995 the title was Extensive wetting
due to roughness and was authored by F. G. Yost, J. R. Michael and E. T. Eisenmann. If you need I will send you a reprint of the paper.
John, Did you ever get any positive responses. Find anything that works? John Hunt} } Hello, } } I am interested on the community input on performing in-situ analysis of solder } to determine composition. We have tried to do this in the past and have not } had acceptable results. We used both multiple areas and multiple points and } averaged the data. } } I realize that this is probably an impossible task with EDS, but would like to } confirm my opinion and obtain input as to any other techniques (micro probe } XRF, etc.) which may provide a non-destructive method for in-situ solder } analysis. } } Thanks, } } John Giles } jegiles-at-space.honeywell.com }
------------------ RFC822 Header Follows ------------------ Received: by smtpmm.space.honeywell.com with SMTP;5 Dec 1995 15:36:22 U Received: from WARF.MSC.CORNELL.EDU by fl51mail.space.honeywell.com with SMTP (1.38.193.5/16.2) id AA03705; Tue, 5 Dec 1995 15:33:34 -0500 Received: from borg.msc.cornell.edu (BORG.MSC.CORNELL.EDU [128.84.249.239]) by warf.msc.cornell.edu (8.6.12/8.6.12) with ESMTP id PAA19651 for {giles_john_e_jr-at-space.honeywell.com} ; Tue, 5 Dec 1995 15:33:58 -0500 (8.6.12/Cornell-MSC-IDA-client for msc.cornell.edu); Tue, 5 Dec 1995 20:33:58 GMT Reply-To: hunt-at-msc.cornell.edu Message-Id: {199512052033.UAA52807-at-borg.msc.cornell.edu} X-Originated-From: borg.msc.cornell.edu X-Msc-Version: IDA Client - AIX
We are contemplating eliminating Osmium Tetroxide as a post fix because of toxicity and disposal concerns. We are interested in documenting morphological changes of the rabbit retina as part of a drug toxicity, efficacy study. Retinas are aldehyde fixed in 2.5% Glutaraldehyde/2.0% Paraformaldehyde solution, followed by Osmium, ethanol dehydration and epoxy embedding. 95% of the blocks will be for light evaluation only. 5% will be selected for TEM low mag wide field. TEM magnification range is 1200-2400X. No intermediate or higher mag's are ever done.
1) by eliminating OsO4, will the contrast and fixation be adversely affected at A) the light level? B) the TEM level (1200-2400x mag)?
if so,
2) is there a suitable alternative to OsO4?
3) at a TEM mag of 1200-2400X, will the differences in contrast and/or fixation be highly noticeable if OsO4 is eliminated?
The 18th International Conference on Cement Microscopy will take place in Houston 21-25 April 1996. You can contact Linda Hills, Construction Technology Labs Inc., 5420 Old Orchard Road, Skokie, IL 60077, USA. Phone: +1 708 965 7500, fax: +1 708 965 6541.
Yours sincerely,
J. B. Bilde-Soerensen Materials Dept. Risoe National Laboratory DK-4000 Roskilde Denmark.
While examining the crystallography of the intermetallic TaRu phase, I discovered that very weak superlattice spots were present in the electron diffraction patterns which were not observable in x-ray powder diffraction patterns, even with high resolution and slow scans. I've heard that this is not uncommon, but was wondering what the reason for this might be. I can imagine a number of possible contributing factors, including an inherent S/N difference, orientation effects, and the non-linearity of film exposures, but what is the best explanation?
Dick Fonda
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 _____________________________________________________________
I thought others in the group might interested in this as well. For replacement photographic bellows, if you have no luck finding OEM bellows, try:
Gortite A and A Manufacturing Co. Inc. 2300 S. Calhoun Rd. New Berlin Wisconsin 53151
414-786-1500 fax: 414-786-3280
They have a whole line of 'photographic' bellows (cameras, enlarger, copy machines, etc.). I just found them, but I haven't tried them yet. Good luck.
(No financial relationship)
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu internet
I am sectioning cod eggs & newly hatched fry embedded in paraffin and preparing them for the Apop Tag DNA stain. I am using silanized slides and drying the slides on a slide warmer overnight and I am still having trouble with losing a significant amount of my specimens after the Apop procedure. Any advice on improving the adhearance of my specimens or a way to decrease the loss of my specimens during the staining procedure would be greatly appreciated.
Contact Diane Nicks at 314 875 5399
or via this e-mail address: Diana_Papoulias-at-nbs.gov
quantitative analysis software , for unix or run on PC ( i need buy) standard for differents minerals and elements ( Au,Ag, As,Fe) ( i need analysis result microprobe of differents minerals i have microprobe cameca MS#46 ( are good) and i need study gold-pip of river
quantitative analysis software , for unix or run on PC ( i need buy) standard for differents minerals and elements ( Au,Ag, As,Fe) ( i need analysis result microprobe of differents minerals i have microprobe cameca MS#46 ( are good) and i need study gold-pip of river
We have not specifically looked at retina, but have on occassion left Osmium out of the soup for various reasons. In general the results are:
1. Reduction in contrast in thin sections (can't comment on thicks) even with heavy Ua/Pb staining.
2. Very poor preservation of lipids.
We also have concerns about Osmium, but I don't know of any suitable substitue. I will be interested in other replies.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Wed, 6 Dec 1995, Fred Hayes wrote:
} We are contemplating eliminating Osmium Tetroxide as a post fix because of } toxicity and disposal concerns. We are interested in documenting } morphological changes of the rabbit retina as part of a drug toxicity, } efficacy study. Retinas are aldehyde fixed in 2.5% Glutaraldehyde/2.0% } Paraformaldehyde solution, followed by Osmium, ethanol dehydration and epoxy } embedding. 95% of the blocks will be for light evaluation only. 5% will be } selected for TEM low mag wide field. TEM magnification range is 1200-2400X. } No intermediate or higher mag's are ever done. } } } 1) by eliminating OsO4, will the contrast and fixation be adversely affected at } A) the light level? } B) the TEM level (1200-2400x mag)? } } if so, } } 2) is there a suitable alternative to OsO4? } } 3) at a TEM mag of 1200-2400X, will the differences in contrast and/or } fixation be highly noticeable if OsO4 is eliminated? } } sections for TEM are post stained in UA/Pb } } Many thanks, } } Fred Hayes } }
If anyone has a Hitachi SEM (or Jeol) with a LaB6 source that would be available for making some comparative pictures, please let me know. Location in southeastern USA, particularly near Atlanta, is preferred but not required. Thanks, Michael Lamvik {mlamvik-at-mcnc.org} MCNC, Research Triangle Park, NC
We are hoping to find some parts for our Philips 201 TEM. We are currently in great need of specimen holders (including both injector rods and tips). We are also low on plate film holders (about 3"x4" in size). We are interested in finding a 70mm camera hook up for roll film; as now we are using only plate film which is quite expensive. We would also be interested in a TV adaptor for the scope. So, if you have these parts please let us know, and contact me at aslamkha-at-hawaii.edu.
} While examining the crystallography of the intermetallic TaRu phase, I } discovered that very weak superlattice spots were present in the electron } diffraction patterns which were not observable in x-ray powder diffraction } patterns, even with high resolution and slow scans. I've heard that this is not } uncommon, but was wondering what the reason for this might be. I can imagine a } number of possible contributing factors, including an inherent S/N difference, } orientation effects, and the non-linearity of film exposures, but what is the } best explanation? } Dear Dick, I don't know the parameters either of the diffraction set-ups or your material; however, with those caviats, since ED can use a small number of unit cells in a selected area, and XD requires more unit cells due to the lower interaction cross-section, if there are changes in the superlattice over distances large compared to the selected area but small compared to the powder grain size, this could account for it. Another possibility is a change in the structure during the preparation of the powder (unless the ED was obtained from individual powder grains). Non-linearity of the film should not affect the presence of the spots--only their intensities. Sat- uration would, in fact, enhance the fainter spots. Yours, Bill Tivol
I need to connect two SCSI-2 cables end to end and am wondering if someone makes a SCSI-2 adapter to do this? The adapter would have two female receptacles to accept the male mini 50-pin connectors of the cables. I have checked all of the usual specialty cable suppliers like Datatek but they could not help. Suggestions? Thanks.
############################################################################# John J. Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Subject: Time: 1:00 PM OFFICE MEMO RE: Xray vs Electron Diffr Date: 12/7/95
At least part of the reason the superlattice spots show up in the electron diffraction patterns, but not in x-ray diffraction patterns, stems from the fact that at small scattering angles the atomic scattering factors for electrons are several orders of magnitude larger than those for x-rays. This matter is discussed more fully in a chapter I wrote on electron diffraction 'way back in 1960 in "Physical Methods in Chemical Analysis", W. G. Berl, Ed. 2nd. edition, p. 620, Academic Press, 1960. W. C. Bigelow (bigelow-at-umich.edu)
Meeting Name: American Association of Feed Microscopists Meeting Dates: June 17 - 21 at Iowa City, Ia Meeting Topic : 43rd Annual Meeting on 17, 18, 19 with topics related to feed microscopy.
Short Course on 19, 20, 21. The short course will teach a person how to I.D. feed ingredient with the microscope. A certificate to those that pass. Meeting Sponsor: AAFM
Contact Person: Marjorie McCutcheon P. O. Box 5246 Charleston, W.V. 25361
I am looking for a source of commercial software to do 3D image reconstruction in a work station type environment (Silicon Graphics, Sun, etc.) using the input from a microscope mounted TV system or, preferably, a slow scan camera. The application involves the use of biological serial sections at approximately 10,000x microscope magnification. It has also been mentioned that the reconstruction could be done using fewer, thicker sections along with the eucentric tilt of the microscope to provide input to such a program.
My feeling is that most folks using work stations have devised their own methods or modified software to suit their unique purposes.
I need the address and cost of the NIST EDS software but don't have a phone number or address. If someone has it please reply to my e mail. Thanks Judy Murphy
Judy Murphy, PhD Dept. of Microscopy San Joaquin Delta College 5151 Pacific Ave Stockton, CA 95207
} 1) by eliminating OsO4, will the contrast and fixation be adversely affected at } A) the light level? } B) the TEM level (1200-2400x mag)? } } 2) is there a suitable alternative to OsO4? } } 3) at a TEM mag of 1200-2400X, will the differences in contrast and/or } fixation be highly noticeable if OsO4 is eliminated?
I sometimes leave out osmium from retina fixation, for LR White embedding. At the low mag you need fixation defects won't be too obvious, but contrast will be poor. I have to stain with UAc/PbC for much longer, use a smaller aperture and print on harder paper. LM isn't affected.
Diana van Driel Dept Ophthalmology Sydney University 2006 NSW, AUSTRALIA
Alternate-Recipient: allowed Auto-Forwarded: prohibited Content-Return: allowed Disclose-Recipients: prohibited Conversion: allowed Importance: normal Priority: normal Sensitivity: Company-Confidential
Several months ago I received a flyer from one of the supply houses announcing a new style of section collection loop that was made by the same process used to make electron microscope grids. The flyer claimed that the design caused the sections to concentrate in the center of the loop and aid transfer to the center of the grid. Now I cannot find the advertisement.
1. Do any of my fellow microscopists remember the advertisement? Can you point me to the correct vendor?
2. Have any of you used these new loops? Do they perform as advertised?
To save bandwidth and prevent those "my mailbox runneth over blues" please respond directly to minter-at-kodak.com and I will post a summary to the newsgroup.
Thanks. -- Best Regards, John
John R. Minter, Ph. D. Phone: (716) 722-3407 Eastman Kodak Company FAX: (716) 477-3029 Analytical Technology Division email: minter-at-kodak.com Rochester, NY 14562-3712
You might try staining with tannic acid/ferric chloride, which we have used as an alternative to OsO4 to preserve antigenicity, etc. TA is added to the fix (must be Na-PIPES or NaPO4 buffer not K or TA precipitates) then you postfix with the same concentration of aqueous FeCl3 (no buffer). This is described in a paper by Overall et al. 1982 Protoplasma 111:134 (used 2% TA in glut/form fix followed by 2% FeCl3, each for 2h), in which older references are cited. Have no idea if it would work well with animal tissues but it brings out all plant membranes and other cell components (i.e. ones that we're interested in!) very well. I haven't used OsO4 for years. We now use only 0.5% TA and FeCl3. If you add up to 10% DMSO to the fixative, penetration is very good (may not do much for membranes, but we're only after the cytoskeleton). UA/Pb staining of sections is fine. More info on TA is given in Mollenhauer and Morre 1987 Histochemistry 88:17
For some immunoEM work I don't use any postfix at all - membranes are fuzzy but UA/Pb stain after gold labelling is OK as long as the Reynolds Pb is at the correct pH.
Hope this helps, Rosemary WHite
____________________________________________________________ Rosemary White __ / Department of Ecology _/ \__/ \ and Evolutionary Biology / \ Monash University / Australia \ Clayton, Victoria 3168 \ ____ / phone 61-3-9905 5670 \_/ \_*_/ fax 61-3-9905 5613 __ email r.g.white-at-sci.monash.edu.au \/
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
There has been some recent discussion about carbon paint used for SEM sample prep:
Make sure it's pure graphite! I've found carbon paints sold for mounting specimens for EDX that are contaminated with phosphorus . Phil Oshel
This is a good example of seemingly similar products not being so similar. If you compare the EDS spectra from a "dab" of the SPI Carbon paint on a spectroscopically pure carbon SEM mount vs. the carbon mount only control spectra, by stretching the imagination, one might see a bit of an Si peak in both scans and in addition, on the carbon paint scan, but not on the control, you again, by really stretching it, some claim to "see" a very tiny indication of Cl. Other than that the paint is "clean". This to me is the acid test of purity.
We have tested other carbon paints in our laboratory over the years. All paints are most definitely not created equal when subjected to the above quality test and there is some variability in purity and also, quality of the brush/cap assembly (which can also release unwanted elements)..
I would be happy to send copies of the original data should anyone want to receive a copy. Send your FAX number with your request.
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: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
I have received a request from Professor S.Maksimov of the Laboratory of electron microscope investigations at Moscow Institute of Electronic Technology Russia (E-mail: lemi-at-mx.iki.rssi.ru) to post a long message to the listserve= r.
Basically his group is looking for collaborators (and sources of funding for a project they are working on). on A3B5 semiconductor compounds.
As a favor, I have edited their long text message and attached an abbreviated copy herein. If anyone is interested in this project or knows of someone that might be can you pass this along. If you want the longer version I'm sure he will be happy to send it to you via Email.
} Please, could You study our information and if it is possible } place it on Microscopy Listserver or give us a council concerning a } place of its location. } } New Mechanism of Dissipative Structure Formation in A(3)B(5). } } We are looking for partners in investigations of regularities of } non-equilibrium ordering (atomic ordering and composition automodula- } tion) in epitaxial layers of the A3B5 compositions: GaAlAs, GaAsP, etc. } We have carried out such investigations beginning with 1978.(S.K. } Maksimov, E.N.Nagdaev, Doklady Akademii Nauk SSSR, (in Russian) 1979, } 245, 1369). } We are interested in receiving of the financial support of the } scientific project from international organization for continuation of } investigations what we have carried out recently (really without any } financial support) and in which we obtained the results allowing to } examine the problem in the new light. At present we have finished the } interpretation of part of these results and prepare them for a publi- } cation (see below the short description of these results). } We are trying to get the financial support in the frames of the } joint research projects of INTAS and RFBR (but, of course we will be } glad any other possibility of a support) and for these purposes are } looking for two research groups in West Europe, which would be } interested in mutual investigations and could participate in them. } Unfortunately,the dead-line of the presentation of the project is 15 } of December. } } Information via Internet } } The information package is also available via: } + World Wide Web on the following sites through CORDIS (the } Community R&D Information Service), INTAS and RFBR: } o CORDIS ([8]http://www.cordis.lu/cordis/) } o INTAS ([9]http://www.ib.be/intas/) } o RFBR ([10]http://www.rfbr.ru/kon/intas/toc.en.html) } } The idea of these investigations (at least, in the theoretical } aspect) arose during the conference "Microscopy of Semiconductor Mate- } rials" which took place at Oxford University on 25-28 March 1991, but } absent of a financial support dragged out its realization.
} Our results. (THIS PART IS EDITED FROM THE ORIGINAL)
} We studied structure and physical properties of GaAlAs epitaxial } layers with a help of techniques: TEM, HREM, TED, reflection ED, ca- } thodoluminescence, etc. In Ga(1-x)Al(x)As/(001)GaAs produced by the } MOCVD technique two types of the dissipative structures competes: the } atomic ordering can be changed by the composition automodulation. At x } =3D 0.2 the ordering direction coincide with the growth direction, but } at x =DA 0.3 it is changed by more traditional that: [010] lying in the } surface plane. Transition from the ordering to the modulation is pro- } voked by breaking of the layer growth mechanism and transition to the } many-center origin (this type of the origin stimulated an acceleration } of the growth). Modulation direction is perpendicular or incline to } the growth direction and does not correspond to models of dissipative } structure formation. Specimens keeping the atomic ordering have only } one line of cathodoluminescence corresponding to an average compositi- } on of the layer but for Ga(0.7)Al(0.3)As (for which structural inves- } tigations showed existence of the modulation)the lines corresponding } to GaAs, Ga(0.3)Al(0.7)As and Ga(0.4)Al(0.6)As are observed. Existence } of modulation is the indubitable phenomenon but its specific features } demands an explanation. Besides the theoretical analysis demonstrates } that an existence of the modulation leads possibly to corrections of } mechanisms of the atomic ordering. } The same dissipative structures are observed for different } technologies and materials and this fact has led us to the idea that a } reason of the non-equilibrium phase transformations must be } fundamental and characteristic for the all A(3)B(5) compounds and does } not connect with separate technologies.
Lots of supporting TEXT DELETED you may get the rest from Maksimov at the address listed below.
} Professor S.Maksimov } } Laboratory of electron microscope investigations } Moscow Institute of Electronic Technology } Russia } E-mail: lemi-at-mx.iki.rssi.ru } } }
The Oxford Instruments Microanalysis Group has just acquired the Microspec Company in Fremont, CA, adding Microspec's WDX400/600 wavelength dispersive (WDX) spectrometers to its range of Link products and systems. There will be no change in the management or staff at Microspec, and as Richard Wolf, President of Microspec, points out to their customers "Only the company name will change". Don Grimes, Microscopy Today
} I need the address and cost of the NIST EDS software but don't have a phone } number or address. If someone has it please reply to my e mail. } Thanks } Judy Murphy
The person in charge of DeskTop Spectrum Analyzer (DTSA) at NIST is Bob Myklebust. The cost is $815 for version 2.0. Call (301) 975-2208 for more information.
Ed Vicenzi tel (609) 258-1464 office Princeton University tel (609) 258-1406 lab Princeton Materials Inst. fax (609) 258-6878 70 Prospect Ave. Princeton, N.J. 08540-5211 vicenzi-at-princeton.edu
We are looking for an alternate company to coat fluorescent TEM screens. We have used Grant Scientific in the past with somewhat satisfactory results but find that the recoated surface does not last. Is there another company or EM supplier that coats TEM screens?
Thanks in advance
---------------------------------------------------------------------------- --------- Karen Vaughn Tel.(904) 392-1184 EM Technician University of Florida Fax.(904) 846-0251 Electron Microscopy Core Laboratory Email. KLV-at-biotech.ufl.edu Interdisciplinary Center for Biotechnology Research http://www.biotech.ufl.edu/~emcl/ 214 Bartram Hall Gainesville, Fl 32611
Does any one have a good protocol or references to techniques localizing oxidases that generate peroxides at the light or electron microscopic level? Thanks in advance.
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
Try APS Technologies, 800-304-9057 They have a variety of SCSI products and have very good service support.
} I need to connect two SCSI-2 cables end to end and am wondering if someone } makes a SCSI-2 adapter to do this? The adapter would have two female } receptacles to accept the male mini 50-pin connectors of the cables. I have } checked all of the usual specialty cable suppliers like Datatek but they } could not help. Suggestions? } Thanks.
R. Howard Berg, Ph.D. Biology Department University of Memphis Memphis, TN, 38152 E mail: bergrh-at-cc.memphis.edu phone: 901-678-4449 fax: 901-678-4457
Message-Id: {199512081615.LAA25190-at-thomas.ge.com} X-Authentication-Warning: thomas.ge.com: daemon owned process doing -bs
I had a similar experience with intermetallic Fe3Al-C prepared by rapid solidification techniques. The samples had an FCC structure in X-ray diffraction but the electron diffraction patterns had weak superlattice reflections. In my case, when the superlattice reflections were used to form a dark field image, small areas within the matrix were imaged. These areas were an ordered phase in a disordered matrix. With different solidification rates, the intensity of the superlattice reflections changed and the volume fraction of ordered regions correspondingly increased or decreased.* I had a lot of help from Joe Horton at Oak Ridge National lab who is an intermetallic, TEM, X-ray wizard. Good luck, Sharon
*S.A. Myers and C.C. Koch, "Electron Diffraction Structure Analysis of Rapidly Solidified (Fe,Ni)3Al-C Alloys",Ultramicroscopy 30 (1989) 193.
Thank you all very much. I needed it very quickly and got the info in a very timely fashion. To respond to someone's comment, I did call NIST but didn't hit the right place. Thanks again, Happy Holidays, Judy Murphy
Are you familiar with the large package of PC software that came out of Caltech (mostly from John Armstrong?) known as CITZAF? It would do what you need, but is not set up for automated work. It can do just about any kind of ZAF correction as well as Bence-Albe, and it is freely available. Let me know and I can send you a copy. Best, Ken Severin
Ken Severin Voice: 907-474-5821 Geology and Geophysics 324 Natural Science Bld. Fax: 907-474-5163 Univ Alaska Fairbanks Box 755780 INTERNET:FNKPS-at-AURORA.ALASKA.EDU Fairbanks AK 99775-5780
We have 2 labs in our hospital that process tissue into methacrylate. One lab uses plastic stubs and a metal adaptor to fit them into the JB-4 microtome. They often experience problems when cutting, the blocks cannot be held firmly enough, resulting in uneven sectioning. The other lab recycles the aluminum stubs, a time consuming task; we use a hammer and single sided razor blade to chop off the specimens for storage and then soak the stubs in water until the methacrylate is soft enough to remove. Has anyone any suggestions for improving: 1) the cutting of sections using the plastic stubs? 2) the method for recycling the aluminum ones????
Thanks in advance
Mark Elliott, PhD UBC-Pulmonary Research Lab Vancouver, BC Canada
} I am looking for a source of commercial software to do 3D image } reconstruction in a work station type environment (Silicon Graphics, Sun, } etc.) using the input from a microscope mounted TV system or, preferably, a } slow scan camera. The application involves the use of biological serial } sections at approximately 10,000x microscope magnification. It has also } been mentioned that the reconstruction could be done using fewer, thicker } sections along with the eucentric tilt of the microscope to provide input to } such a program. } Dear Dennis, If you contact Joachim Frank, he can tell you about the avail- ability of the SPIDER image processing system. I think it can meet your needs since we use it here to do 3D reconstruction by serial sections, thich-section contouring, and other methods. Both Fourier methods and weighted back-projection are used, and projection onto convex sets is also. Once you have a computer file, the program doesn't care whether the data are from film, TV or SSCCD; you will, of course, have to provide the frame-grabber or storage buffer which converts the video image into a digitized file. You can contact Joachim Frank at joachim-at-wadsworth.org or look at the web page "http://www.wadsworth.org". Yours, Bill Tivol
} I am looking for a source of commercial software to do 3D image } reconstruction in a work station type environment (Silicon Graphics, Sun, } etc.) using the input from a microscope mounted TV system or, preferably, a } slow scan camera. The application involves the use of biological serial } sections at approximately 10,000x microscope magnification. It has also } been mentioned that the reconstruction could be done using fewer, thicker } sections along with the eucentric tilt of the microscope to provide input to } such a program.
The following is NOT a commercial.
If you accept manual tracing of interesting structures on an overlay-layer of imported tif images, you might use 3D - T.O.P. XW (of IMATEC, Neufahrn, Germany) running on SGI (IRIX 5.3). It has powerful interactive visualization capabilites (realtime rotation and zooming in wireframe mode), good reconstruction and shading caps, but so far rudimentary import caps only. A new tracer/import module for different file format import (and autotrace features) is announced by early 1996.
I am PhD student at Kaunas University of Technology. Currently I am working in Research Centre "Vibrotechnika". Our research work cover acoustic waves investigation. We are looking for some low priced Atomic Force Microscope for further investigations. This microscope can be used and not new. I will be really thankful to everybody who will send me any proposal.
Address Mindaugas Rackaitis Mickeviciaus 11 3000 Kaunas, Lithuania e-mail: vibro-at-vb.ktu.lt
Which laboratory in the Netherlands can help me? I am urgently looking for a HBO-50W/3 DC lamp. The supplier needs approx. 4 weeks before they can deliver the lamp while we are using the lamp for 40 hrs a week. Can we buy one from you, or borrow, please contact me.
Winfried Leeman TNO Nutrition and Food Research Institute Toxicology Division Phone (+31) 030 - 694 41 44 Direct phone (+31) 030 -694 44 97
I have used JB-4 quite a bit and also (due to the high cost of the metal chucks), switched to the plastic ones. I have really not had too much dificulty in making the switch,although I have occationally used a pair of pliars to secure them into the metal adaptors.
The best way to recycle the aluminuim ones is to secure them in a vise, and use a hammer and straight screwdriver at a 45 degree angle at the base. (be sure and use eye protection) This method also relieves frustration.
Good Luck, Kathy Walters U of Iowa
On Fri, 8 Dec 1995 MELLIOTT-at-prl.pulmonary.ubc.ca wrote:
} JB-4 Specimens Using Metal or Plastic Stubs } } We have 2 labs in our hospital that process tissue into methacrylate. } One lab uses plastic stubs and a metal adaptor to fit them into the } JB-4 microtome. They often experience problems when cutting, the } blocks cannot be held firmly enough, resulting in uneven sectioning. } The other lab recycles the aluminum stubs, a time consuming task; we } use a hammer and single sided razor blade to chop off the specimens } for storage and then soak the stubs in water until the methacrylate } is soft enough to remove. } Has anyone any suggestions for improving: } 1) the cutting of sections using the plastic stubs? } 2) the method for recycling the aluminum ones???? } } Thanks in advance } } Mark Elliott, PhD } UBC-Pulmonary Research Lab } Vancouver, BC } Canada
George Smith from Union College asks for help to refurbish his critical point dryer. He has a Balzers CDP 010. It has a small leak in the line somewhere, plus the top of the bomb is busted and needs to be replaced or repaired. In addition, George is looking for an oil diffusion pump for a Hitachi Vacuum evaporator. If you have any leads contact:
George Smith Biology Union College Schenectady, NY 12308 (518) 388-6241
In response to klv-at-biotech.ufl.edu (Karen L Vaughn), seeking a company that recoats fluorescent TEM screens, Ernest F. Fullam, Inc does TEM screen recoating and has for many years. Please contact us for pricing and more information. Dianne Fullam Ernest F. Fullam, Inc. Phone: (518) 785-5533 FAX: (518) 785-8647 E-Mail: dbf-at-fullam.com
If you have enough of the metal stubs, don't bother breaking off the methacrylate. Just hold them in water for a week or so, then cut off the block. Then soak some more until you can push out the 'tail'.
Regards, Glen
On Fri, 8 Dec 1995 MELLIOTT-at-prl.pulmonary.ubc.ca wrote:
} JB-4 Specimens Using Metal or Plastic Stubs } } We have 2 labs in our hospital that process tissue into methacrylate. } One lab uses plastic stubs and a metal adaptor to fit them into the } JB-4 microtome. They often experience problems when cutting, the } blocks cannot be held firmly enough, resulting in uneven sectioning. } The other lab recycles the aluminum stubs, a time consuming task; we } use a hammer and single sided razor blade to chop off the specimens } for storage and then soak the stubs in water until the methacrylate } is soft enough to remove. } Has anyone any suggestions for improving: } 1) the cutting of sections using the plastic stubs? } 2) the method for recycling the aluminum ones???? } } Thanks in advance } } Mark Elliott, PhD } UBC-Pulmonary Research Lab } Vancouver, BC } Canada }
} I need to connect two SCSI-2 cables end to end and am wondering if someone } makes a SCSI-2 adapter to do this? The adapter would have two female } receptacles to accept the male mini 50-pin connectors of the cables. I have } checked all of the usual specialty cable suppliers like Datatek but they } could not help. Suggestions? } Thanks. } } ############################################################################# } John J. Bozzola, Director } Center for Electron Microscopy } Southern Illinois University } Carbondale, IL 62901-4402 } U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ############################################################################# } } }
John -
Based on painful personal experiences you may not want to do this. First there is a max length for your old run-of-the-mill SCSI-2 cables, its about 6 ft. If the combined cables that make up a classic SCSI-2 chain are longer than that you might experience problems.
My advice is to find a cable that has the correct ends and is as short as possible. Don't try to save a few $'s here, it just ain't worth it!
Thanks to everyone who responded to my request for information regarding 3D Image Reconstruction Software for Workstations. I have included all the responses I have received thus far (with minimal editing). Each response is separated with a line. Other interested parties might find something useful here.
Regards, Dennis
Summary follows:
NASA's biotechnology division released a 3D recon. package specifically for EM and confocal work about 3 or 4 months ago. It was for US use only and was apparently available free of charge if you could justify its use (and were a US citizen). Perhaps you could contact them regarding this. It was " advertised" on the WWW NASA pg.
Universal Imaging have a package called "metamorph" that I have'nt looked at (asked for more info. and did'nt get a response) that claims to do 3D recon. as well as photomontage work that can integrate images from a CCD camera. It supposedly has a number of other abilities and is available from Universal Imaging Corp., 502 Brandywine Parkway, West Chester, PA 19380.
There is also a SW package called VoxalView (sp.) from Vital Images that does 3D recon. It is, however, quite expensive (} $10K). I've asked around about it and been told it is not so good for some specific applications.
We currently have a package that has been custom designed for our purposes (3D recon. from 35 mm EM negs. with automated alignment to do volume and area measurements of smooth muscle cells and associated nerves). We have'nt gotten as far as publishing with it yet, however when we have we intend to have it available for general distribution and use. According to the guy thats developing the program there are a number of useful bits on the WWW, but I think that's for people who have the time and know how to find them (as I certainly would'nt know where to start).
Regards,
Shaun Sandow, Division of Neuroscience, John Curtin School of Medical Research, Australian National University, ACT 0200
I have been associated with a couple labs trying to solve this problem, and we at NRL have also investigated the problem for an upcoming program. In a nutshell, there do exist commercial programs out there, particularly geared towards the medical community. However, they typically have a low resolution and cannot handle the more complex structures often seen in materials science. In every case I know of, this has led to (at least an attempted) development of the actual software. Unfortunately, the research group which appeared to be heading this effort, at least in the area of solid-solid phase transformations (headed by Gary Shiflet of UVa) just lost much of their computer support and no longer has a feasible system (as far as I know). We will be working to develop our own system probably within the next 1/2 year because of this.
For the application you describe, the commercially available software may have sufficient resolution. We have not checked out that software, but Dr. Shiflet did before he started developing their own system. You might want to contact him at gjs-at-gjssgi.ms.virginia.edu for further information.
Dick Fonda [Naval Research Lab, Washington, DC] _______________________________________________________________________
VayTek, Inc. 305 West Lowe, Suite 109 P.O. Box 732 Fairfield, Iowa 52556 515-472-2227 FAX: 515-472-8131
Christopher MacLean, Ph.D.
vaytek-at-ins.infonet.net
Software for PC's (DOS/Windows 3.11, and Windows NT), Unix, and Mac's. Their main software for 3D reconstruction is called VoxBlast, they also have assisant software for aligning 2D sections, and they have deconvolution microscope controlling software as well. All of which is designed to be intergrated with each other and Image Pro Plus.
I have tried their demo software (available on line at ftp://128.255.88.195, User: vbdemo, password: 5UZY2fCn [case sensitive]), and I have had demos done for me but I have yet had the funding to get my own copy of the software. The technical support and sales reps are friendly, knowledgable, and very willing to help.
A second set of software for Unix (and PC's?) is VoxelView, but I do not have the contact info available for them.
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu _______________________________________________________________________
I am using Spyglass, which is now distrubuted by Fortner Research (tel: 800 252-6479 or http://www.langsys.com/langsys.
I am analyzing serial sections obtained by SEM and run the application on a Mac (good enough for this purpose). However, Spyglass is also available for UNIX platforms. Excellent software and easy to use (I never used a manual). I typically have 20 sections, each 640*512 pixels and they are rendered rapidly.This software is only for data reprssentation and not for data anaysis. It would be desirable to be able to extract stereological features from any 3D data set, however, this is not build in.
I hope that this helps. Please let me know if you come across other sources for this applications.
Hasso Weiland Senior Scientist Aluminum Company of America Alcoa Technical Center Alcoa Center, PA 15069 412-337-3133 weiland_h-at-atc.alcoa.com ______________________________________________________________________
Have you seen Voxel View? We used it at Texas A&M to reconstruct confocal sections on a SGI system. I think it would fit your app, but like most workstation software - it's expensive.
Good Luck,
James C. Long Manager/Materials Analysis Lab Electrosource, Inc. 512-445-6606 jlong-at-bga.com _______________________________________________________________________
Try the John Steven Groug at Playfair in Toronto.
Contact Judy Trogadis judy-at-CAMTWH.ERIC.ON.CA or John, john-at-camtwh.eric.on.ca
He has written the book on this type of thing which is very different that using confocal input data because TEM sections get distorted and misaligned before they are recorded.
Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU _____________________________________________________________________
If you contact Joachim Frank, he can tell you about the availability of the SPIDER image processing system. I think it can meet your needs since we use it here to do 3D reconstruction by serial sections, thich-section contouring, and other methods. Both Fourier methods and weighted back-projection are used, and projection onto convex sets is also. Once you have a computer file, the program doesn't care whether the data are from film, TV or SSCCD; you will, of course, have to provide the frame-grabber or storage buffer which converts the video image into a digitized file. You can contact Joachim Frank at joachim-at-wadsworth.org or look at the web page "http://www.wadsworth.org". Yours, Bill Tivol ________________________________________________________________________
Your're right-most individuals with workstations have developed their own custom software. Jill Gemmill, at the Neurobiology Research Center at University of Alabama at Birmingham, Birmingham, Al (205-934-7111) developed an excellent 3-D reconstruction package on a Silicon Graphics Workstation. The software was used to perform three dimensional serial reconstruction from images generated by photographs of serial sections in an electron microscope. Granted the interface to capture images directly to the workstation via slow scan camera is not there, but the reconstruction software that Jill developed is super. I cannot find an E-mail address for her, but above is a direct line to her facility. If anything, she might be able to help you out a bit.
Best Regards
Kevin Kevin McCarthy Assistant Professor Department of Cell Biology Digital Imaging Microscopy Facility University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029 "Seeing the World Through Different Eyes" ________________________________________________________________________
Mayo Clinic puts out a software package that can do almost anything you want it to, however the onsite cost is around $60,000.00.
I have a friend and College at our Beckman Research Institute that is going to teach this software to me. Her name is Janet Sinn-Hanlon, and her email address is:
Janet-at-delphi.beckman.uiuc.edu
* Janet works only part time, so it may take her a few days to get back to you. Tell her I sent you, she would be glad to discuss the software with you. It could be an individual purchase would be much less expensive.
Good Luck,
Lou Ann Miller Microscopic Imaging Lab College of Vet. Medicine University of Illinois 2001 S Lincoln Ave Urbana,Illinois 61801 217-244-1566 lmiller-at-ux1.cso.uiuc.edu
Microscopy Home Page: http://www.cvm.uiuc.edu/MicImagLab/MicImagLab.html
Personal Home Page: http://www.cvm.uiuc.edu/HomePages/LouAnnMiller/Homepage.html ________________________________________________________________________
The following is NOT a commercial.
If you accept manual tracing of interesting structures on an overlay-layer of imported tif images, you might use 3D - T.O.P. XW (of IMATEC, Neufahrn, Germany) running on SGI (IRIX 5.3). It has powerful interactive visualization capabilites (realtime rotation and zooming in wireframe mode), good reconstruction and shading caps, but so far rudimentary import caps only. A new tracer/import module for different file format import (and autotrace features) is announced by early 1996.
E.A. Fischione Instruments, Inc is seeking a highly skilled and motivated individual for a senior machinist/instrument maker position. Duties will include working with the engineering staff throughout the design and development of innovative electron microscopy accessories including the fabrication and intricate assembly of ultra-precise mechanical/electro- mechanical components. Additional activity will be in the support of production utilizing state-of-the-art CNC turning and vertical machining centers. The candidate must be able to read engineering drawings. Knowledge of manufacturing practices for vacuum usage is a plus.
E.A. Fischione Instruments, Inc. was founded in 1966 and specializes in the development and manufacture of TEM specimen preparation devices. The company is located in Export, PA, approximately 23 miles east of the city of Pittsburgh.
Resumes should be sent to: Paul E. Fischione, President E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632
Has anyone had dealings with Imidazole + Osmium tetroxide post fixation methods.... And if they have, have they experienced very dark, electron dense circular deposits around the outside of the tissue? These deposits are not within the tissue so could this be a precipitate of some sort with some of the other chemicals used in processing? Does Imidazole react with aldehydes/solvents to cause this type of thing?
Any comments from anyone with experience in the use of the above technique would be appreciated!
Regards,
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
E.A. Fischione Instruments, Inc. is seeking a highly motivated individual for an applications engineering position. Duties will be to provide both input for new product developments and the field support of existing products. An extensive knowledge of TEM specimen preparation techniques is essential. The candidate must be well versed in both theoretical and operational aspects of Transmission Electron Microscopes and must be able to conduct routine TEM maintenance.
The ideal candidate should possess a Ph.D. in Materials Science with a B.S. or M.S. in either engineering or physics. As an applications engineer a great deal of activity will focus on interactions with both existing and potential customers. Technical publications including techniques and results developed with Fischione instrumentation will be encouraged. The position requires excellent verbal and written communication skills and a willingness to do extensive travel.
E.A. Fischione Instruments, Inc. was founded in 1966 and specializes in the development and manufacture of TEM specimen preparation devices. The company is based in Export, PA, approximately 23 miles east of the city of Pittsburgh.
Resumes should be sent to: Paul E. Fischione E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632
Message-Id: {199512121243.HAA27970-at-aretha.jax.org} X-Sender: meh-at-aretha.jax.org X-Mailer: Windows Eudora Version 1.4.4 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
I am looking for vendors that carry glass strips for making microtome knives.
Specifically,
4inch x 2inch x 100mm 25mm x 6.35mm x 400mm
names and phone numbers would be appreciated,
thank you!
Margaret E. Hogan, PhD, Supervisor Biological Imaging Service (Histology, Electron Microscopy, Image Analysis) The Jackson Laboratory 600 Main Street Bar Harbor, Maine 04609 (207) 288-6191 (207) 288-5079 FAX meh-at-aretha.jax.org
Message-Id: {199512121329.IAA00651-at-aretha.jax.org} X-Sender: meh-at-aretha.jax.org X-Mailer: Windows Eudora Version 1.4.4 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Sorry, I just noticed my mistake on this message, the message and dimensions should read as:
I am looking for vendors that carry glass strips for making microtome knives.
Specifically,
4inch x 2inch x 10mm 25mm x 6.35mm x 400mm
names and phone numbers would be appreciated,
thank you!
Margaret E. Hogan, PhD, Supervisor Biological Imaging Service (Histology, Electron Microscopy, Image Analysis) The Jackson Laboratory 600 Main Street Bar Harbor, Maine 04609 (207) 288-6191 (207) 288-5079 FAX meh-at-aretha.jax.org
Dear sir: I would like to subscribe to the microscopy newsgroup. Please send any particulars. George W. Smith Biological Sciences Union College Schenectady, NY 12308 518-388-6245 518-388-6429 fax smithg-at-gar.union.edu
HELP. I'VE BEEN ASKED TO DO A ROTAVIRUS STUDY ON STOOL SPECIMENS. THE PROTOCOL CALLS FOR INCUBATING THE SPECIMEN WITH ROTAVIRUS ANTIBODY, CENTRIFUGING AT 12,000 RPM FOR 90 MINUTES, THEN TREATING THE SPECIMEN AS A NEGATIVELY STAINED SPECIMEN. OUR FASTEST CENTRIFUGE IS 13,000 RPM. HAVE ORDERED A POLYCOLNAL ANTIBODY (HUMAN) TYPES 1,2,3,4,8. QUESTIONS: CAN A SHORTER CENTRIFUGE TIME BE USED? CONCERNED ABOUT HEAT BUILD UP, ETC. BESIDES FLAMING ( NOT ALLOWED AT THIS HOSPITAL), WHAT IS AN EFFECTIVE WAY TO STERILIZE THE FORCEPS/ PREVENT CARRYOVER? BEST WAY TO FILTER/ EXTRACT VIRUS FROM STOOL PARTICULATES? COMMENTS FROM ANYONE WITH EXPERIENCE WITH THIS OR SIMILAR PROCEDURES WOULD BE APPRECIATED. BECKY GARRISON EM LAB PH 904-549-4218 PATH DEPT/UMC FX 904-549-4290 JACKSONVILLE FL
We are in need of a user tracking program that will monitor the use of a multi-user image analysis system which runs on OS/2. Does anyone have a idea where to find such a program? Any help is appreciated.
Thank you in advance, Davin
If you are running on Novell, you can try a program called LAN Record.
We are in need of a user tracking program that will monitor the use of a multi-user image analysis system which runs on OS/2. Does anyone have a idea where to find such a program? Any help is appreciated.
Thank you in advance, Davin
Davin Jutila Research Imaging/Flow Cytometry Facility Southern Illinois University School of Medicine Springfield, IL 62794-9230 E-mail: djutila-at-wpsmtp.siumed.edu
Perchance, does anyone have the phone number for Univeristy Micorfilms of Ann Arbor MI?
Bob
Robert R. Wise, PhD Department of Biology University of Wisconsin Oshkosh Oshkosh, WI 54901 (414) 424-3404 tel (414) 424-1101 fax wise-at-vaxa.cis.uwosh.edu
Although I haven't tested imidazole+OTO per se, I have tried a variety of (osmium ligands+ OTO) in developing freeze substitution media, using acetone as the solvent. Visually dark products in these samples tend to be leached out during acetone-only rinses, so I keep the ligand in post-OTO solutions, in which case the embedded tissue appears darker. This work is in progress, sorry I can't offer a more complete analysis.
} Richard Lander asks: } } Has anyone had dealings with Imidazole + Osmium tetroxide post fixation } methods.... And if they have, have they experienced very dark, electron } dense circular deposits around the outside of the tissue? These deposits } are not within the tissue so could this be a precipitate of some sort with } some of the other chemicals used in processing? Does Imidazole react with } aldehydes/solvents to cause this type of thing?
R. Howard Berg, Ph.D. Biology Department University of Memphis Memphis, TN, 38152 E mail: bergrh-at-cc.memphis.edu phone: 901-678-4449 fax: 901-678-4457
Does anyone have a reliable way to make carbon-coated grids for high resolution work? I have tried floating carbon films off mica-- they seem to fall apart before I can pick them up on grids. Any tips or suggestions?
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
Margaret Hogan Wrote: } Subject: glass for knives (correction)
I am looking for vendors that carry glass strips for making microtome knives. Specifically,
a) 4inch x 2inch x 10mm and b) 25mm x 6.35mm x 400mm } } names and phone numbers would be appreciated, thank you!
Please specify whether you are seeking to make thin sections for TEM or thick sections for LM as the choice of glass will depend on which kind of sections you are wanting to make.
Both types of glass are available from SPI Supplies as well as several of the other major suppliers of EM consumables.
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: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
The usage of slow-scan cameras is increasing very much, so there is a need to transfer those images between different programs. Since those slow scan CCDs produce grayscale images with more than 8 bits/pixel ( normally 12 or 14 bits/pixel ) there is a file format required that supports data up to 16 bits/pixel.
I downloaded the Aldus Tiff specification Revision 6.0, Final Q, june 3, 1992 that seems to me to be the latest one. In this specification I found the tag 258 (102H) BitsPerSample that contains the bits per pixel. It would be easy to write here a file with BitsPerSample=16 but in the specification is explicitly written "Allowable values for Baseline TIFF grayscale are 4 or 8, allowing 16 or 256 distinct shades of gray."
So what is the use of a program that could write such a file if there is no other (different) program that is able to read this file. (See also the discussion here about "different" TIFF formats ).
Has anyone experience how to transfer 16 bit images in a file format that is known to a couple of image processing or graphic programs ? I am not fixed to TIFF.
At moment I either use 8bit TIFF with previously rescaled data ( that means loss of information) or raw format (without any header, etc.). But also the raw format is only supported by very few programs and you have the big/liddle endian problem.
My thanks for the many messages and helpful suggestions about a=20 replacement for halocarbons. I am grateful too for the gloating=20 meterological information from Australia (25-30C is obscene at this time=20 of year). Holiday greetings from Cambride where the snow has been replaced by gale=20 force fog.
E.A. Fischione Instruments, Inc is seeking a highly skilled and motivated individual for a senior machinist/instrument maker position. Duties will include working with the engineering staff throughout the design and development of innovative electron microscopy accessories including the fabrication and intricate assembly of ultra-precise mechanical/electro- mechanical components. Additional activity will be in the support of production utilizing state-of-the-art CNC turning and vertical machining centers. The candidate must be able to read engineering drawings. Knowledge of manufacturing practices for vacuum usage is a plus.
E.A. Fischione Instruments, Inc. was founded in 1966 and specializes in the development and manufacture of TEM specimen preparation devices. The company is located in Export, PA, approximately 23 miles east of the city of Pittsburgh.
Resumes should be sent to: Paul E. Fischione, President E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632
Kristof KOVACS University of Veszprem, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684
To: psphicas-at-pipeline.com, Microscopy-at-Sparc5.Microscopy.Com
} Does anyone have a reliable way to make carbon-coated grids for high } resolution work? } I have tried floating carbon films off mica-- they seem to fall apart } before I can pick them up on grids. } Any tips or suggestions?
Dear Phil and all other interested people,
The method we use here is as follows (as learnt from our TEM technicians): 1) Place a drop of washing up liquid or teepol on a glass slide and wipe it all over with some tissue paper wiping most of it off in the process (leaving the slide sightly sticky). 2) Form a thin carbon film on this slide in a carbon coater. 3) Score the carbon film with a razor blade or scalpel to suitable size pieces. 4) Float off the pieces of carbon film with water and catch with copper grids. I usually prefer 200 mesh grids and it works fine every time. I use them to support small ceramic particles in the TEM and they usually behave very well up to at least 250k magnification.
Hope this helps
Ian MacLaren, IRC in Materials for High Telephone: 0121 414 3447 Performance Applications, FAX: 0121 414 3441 The University of Birmingham, email: I.MacLaren-at-bham.ac.uk Birmingham B15 2TT, England
I've used two ways to fabricate carbon coated grids.
Plan A. Much as you have done. Generally it seems better to make the carbon film on the mica surface quite thin, something around 10nm (sorry my only monitor is mounted in a freeze-fracture machine so I can't more than speculate on how thin). Score the resulting film into about 4mm squares, float on dH2O, and pick them up on flamed grids. Blot at the edge to dry.
Plan B. Make conventional plastic film coated grids, by whatever means are at your disposal, and pick them up from above on clean paper. Allow to dry then carbon coat. Put the coated grids (film side up) into a chamber made from (e.g.) a glass petri dish ca. 90mm in which there is a pad of filterpaper soaked with a solvent of the film plastic. I usually let them sit over night but the plastic is likely gone and diluted into the solvent more quickly than that. If you want them real clean you can repeat with a fresh chamber assembly. Also makes really nice holey carbon if you start out with glycerol doped plastic.
cheers, John heckman-at-pilot.msu.edu
} Does anyone have a reliable way to make carbon-coated grids for high } resolution work? } I have tried floating carbon films off mica-- they seem to fall apart } before I can pick them up on grids. } Any tips or suggestions? }
E.A. Fischione Instruments, Inc. is seeking a highly motivated individual for an applications engineering position. Duties will be to provide both input for new product developments and the field support of existing products. An extensive knowledge of TEM specimen preparation techniques is essential. The candidate must be well versed in both theoretical and operational aspects of Transmission Electron Microscopes and must be able to conduct routine TEM maintenance.
The ideal candidate should possess a Ph.D. in Materials Science with a B.S. or M.S. in either engineering or physics. As an applications engineer a great deal of activity will focus on interactions with both existing and potential customers. Technical publications including techniques and results developed with Fischione instrumentation will be encouraged. The position requires excellent verbal and written communication skills and a willingness to do extensive travel.
E.A. Fischione Instruments, Inc. was founded in 1966 and specializes in the development and manufacture of TEM specimen preparation devices. The company is based in Export, PA, approximately 23 miles east of the city of Pittsburgh.
Resumes should be sent to: Paul E. Fischione E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632
Kristof KOVACS University of Veszprem, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684
Your request for glass for knives has been forwarded to me by the Pres. of SPI. We can offer you the 2 sizes you have inquired about. The exact sizes in metric are as follows: 101mm x 50.8mm x 10mm , selling price is $121.90 per box of 30 and 25.4mm x 6.4mm x 400mm, selling price is $45.00 per box of 20
Since these are not stock items for SPI, we will require a minimum purchase of 5 boxes of each size. You need only buy one size (5 boxes) you are not required to buy both sizes. Delivery will be about 3-4 weeks.
If you have nay questions, you may contact me by e-mail or use our toll- free # 1-800-242-4SPI.
I would NOT RECOMMEND the use of Teepol or any other commercial washing-up liquid for the preparation of carbon films for ANALYTICAL studies using EDX or EELS - especially with biological specimens where one is interest- ed in serious elemental quantitation! Peter Ingram SEND
I have a user interested in obtaining baskets and wax baths for the AO/Leica TP 8000 tissue processor (catalog #8004 for the basket; #8005 for the wax bath).
If you have some of these items and wish to part with them , please contact Phil Langlais directly at planglai-at-sunstroke.sdsu.edu to discuss terms
Happy Holidays and Best Wishes for the New Year!
steve
--------------------------------------------------------------------------- ------------------------------------------------------------------------------ Dr. Steve Barlow EM Facility/Biology Dept. San Diego State University San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu
Our lab routinely processes human and animal stools for viral diagnosis, and we often use IEM in one form or another. If you plan to agglutinate the virus with specific antibody prior to applying it to the grid, you do not need such extended centrifugation times if you are centrifuging in 1.5 ml "microfuge" tubes. We use a speed of 12 - 13K (about 11,000 xg) for only 5 minutes, and find this to be quite adequate. Another approach which is worth trying is to coat the grid with antibody first (float grid on drop of serum for about 30 min., then blot dry), then apply the stool suspension. After incubating for 30 min., lightly rinse the grid to remove unbound particulates, then negative stain. All that remains is bound, agglutinated virus. This approach is useful for viral titer quantitation. For sterilization, dipping forceps in a 50% clorox solution is more than adequate (at least its acceptable to the CLIA inspectors). We prepare stool suspensions by combining equal parts of stool and DI water, mixing, then clarifying by centrifugation on a microfuge at 13,000 rpm (11,000 xg on our machine) for 2 minutes.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia
EMAS'96 2nd Regional Workshop on Modern development and applications in Microbeam Analysis Organised by:=20 European Microbeam Analysis society in co-operation with Hungarian Microscopical Society Kossuth Lajos University, Debrecen
International Scientific and Organising Committee
J=E1nos L=E1b=E1r (Chairman) Hungary, Luc Van't dack Belgium, Johann= Wernisch Austria, Peter Karduck Germany, Patrick Nicholson U.K, Antonin Simunek Czech Republic, Csaba Cserhati Hungary, Kristof Kovacs Hungary, Peter Nagy Hungary
Just wanted to post my thanks for all the responses I received to my query on Video Archiving Systems for our CD measurement SEM. It was a huge help to hear from other folks who are currently utilizing these types of systems.
The Dow Chemical Company Analytical Sciences 1897E Building Midland, MI 48667
Contact: John Blackson
A R&D position in analytical transmission electron microscopy (ATEM) is available in Dow's Surface Analysis, Microscopy and Xray Group. The applicant should be a self motivated individual having an advanced degree or experience in either ceramics, metallurgy or material science. Candidate must be well versed in electron microscopy. Experience in electron diffraction, energy dispersive spectroscopy, PEELS and sample preparation is desirable. Other areas of competence could include: FEG- SEM, high resolution TEM (HRTEM) and STEM. The position involves the use of state-of-the-art equipment to solve complex industial problems. The group is equipped with: 1 HRTEM, 1 ATEM, a dedicated STEM with EDS and PEELS, 1 CTEM, 2 FEG-SEM's, Enviromental SEM, Analytical SEM, 2 Electron Probes, 1 SAM, 2 XPS units, Scanning Probe Microscopies, 1 HREELS, Confocal laser light microscope, many other light microscopes, full metallographic capabilities, and a variety of x-ray diffractometers.
This position is also being posted on other mail reflectors and newsgroups. 12/7/95
Center of Microanalysis Department of Materials and Nuclear Engineering A. James Clark School of Engineering University of Maryland College Park, MD 20742
The position of Facility Operator of the Center of Microanalysis is vacant, and applications are invited. The person filling this position reports to the Director of the Center. Interested individuals are invited to apply by submitting a letter of interest, resume and list of references (at least three) to the address below. For optimum consideration, applications should be received by February 1, 1996.
Duties of the position include: training and assisting users in operating equipment and preparing samples, routine maintenance of equipment, interfacing with service engineers to resolve major equipment problems, working with Maryland companies to solve microanalytical problems, and coordinating schedules and operations.
The ideal candidate should have the following characteristics: At least a BS degree in a field relevant to microanalysis, knowledge of SEM, EDS, WDS, FTIR, AFM and sample preparation and skills in personal interactions, computer operation, report writing, coordinating schedules and ordering and maintaining materials and supplies.
Submit applications to:
Chairman, Search Committee Department of Materials and Nuclear Engineering A. James Clark School of Engineering University of Maryland College Park, MD 20742
THE UNIVERSITY OF MARYLAND AT COLLEGE PARK IS AN EQUAL OPPORTUNITY EMPLOYER
} I downloaded the Aldus Tiff specification Revision 6.0, Final Q, june 3, 1992 } that seems to me to be the latest one. } In this specification I found the tag 258 (102H) BitsPerSample that } contains the } bits per pixel. It would be easy to write here a file with BitsPerSample=16 but } in the specification is explicitly written "Allowable values for Baseline TIFF } grayscale are 4 or 8, allowing 16 or 256 distinct shades of gray." } } So what is the use of a program that could write such a file if there is no } other (different) program that is able to read this file. (See also the } discussion here about "different" TIFF formats ). } } Peter Sparlinek } 100115.66-at-compuserve.com
If you read the fine print in the TIFF spec, you will find that a TIFF reader (your program) is expected to be able to handle TIFF images with a BitsPerSample of 4 or 8. Most good TIFF readers will also handle 16. Very good TIFF readers will handle BitsPerSample=n where n can range from 1 to big. A good example of this is if your hardware is a 12 bit ADC. Why waste 4 bits storing the image as a 16 bit TIFF when all you need is 12. For our software packages, we decided that for the best cross-program compatibility in reading our TIFF files, if our bit usage was greater that 8, we would use 16. Not as efficent in raw storage but cross-program compatibility was more important. When you read the TIFF spec remember that the original TIFF was designed for visual (look at it) images, not scientific images (crunch the numbers). It was just flexible enough to applied to scientific images (how about BitsPerSample=32 and the data type is 32 bit floating point -- check the TIFF spec, a valid TIFF image but I don't think many programs could read it).
Scott D. Davilla 4pi Analysis, Inc. 3500 Westgate Drive, Suite 403 919 489-1757 (tel) Durham, North Carolina 27707 919 489-1487 (fax)
YOU MIGHT HAVE RECEIVED SOME JOB OPENING ANNOUNCEMENTS FROM E.A. Fischione Instruments, Inc. OR MAYBE SOME OTHER MAIL ORIGINATED BY SOMEONE ELSE, AND ALSO HAVING MY NAME AT THE END.
BELIEVE ME: I HAVE NOT BEEN INVOLVED IN THESE POSTINGS. HERE'S THE TRUTH:
THE SERVER I WAS CONNECTED TO IS ABOUT TO SHUT DOWN OPERATION AND IS FORWARDING ALL MY CORRESPONDANCE TO MY NEW E-MAIL ADDRESS. WE HAVE OUR OWN PROBLEMS WITH THIS SWITCH-OVER BUT APPARENTLY THE SYSOP OF OUR SERVER DID AGAIN SOMETHING WRONG, AND POSTED SOME OF MY INCOMING MESSAGES AS OUTGOING TO VARIOUS ADDRESSES, INCLUDING THIS LISTSERVER. I DON'T KNOW WHAT SHOULD I EXPECT IN THE FUTURE, BUT PLEASE ACCEPT MY APOLOGIES FOR ANY INCONVENIENCE.
Kris
Kristof KOVACS University of Veszprem, Central Laboratory P.O.Box 158, Veszprem, HUNGARY H-8201 Phone: +36-(88)-421-684
A few questions about why you plan on doing these experiments. Do you need only to ID rotavirus, ie. for clinical settings? or do you want to observe the structure of the virus? In either case, mixing the virus and antibody together will end up with large aggragates of virus and identification will be harder. (unless you have the proper ratio of virus:ab) The method of adding ab to a grid then floating on top of a viral suspension works very well (this is called immunosorbent IEM). My recommendations for a stain would be 1% ammonium molybdate and not PTA because it has been shown that PTA can be detrimental to viral particle structures. Another method to consider is not an immuno procedure, one called pseudoreplication. Here a drop of 10% viral solution (which has been centrifuged) is placed on a square of 2-4% agar, allowed to dry, a drop of parlodion solution is placed on agar spread around and tipped on side to drain and dry. The agar plug (on a microscope slide) is then tilted into a dish of stain, upon which the film is floated off, a grid is placed on top of film and then the grid/film is picked up with the use of a small brass rod (2 cm in length and 5mm dia) and forceps. Excess stain is drained and dried then examined. The advantage of this procedure is that any and all viruses are ID'ed and not just a single one. If you need more info, please write or call.
Best of luck, Ed Calomeni Dept Pathology Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu 419-381-3484
Does anyone have any suggestions for fixation and embedding small crustaceans (clam shrimp) about 4mm in lenght?. I'm concerned about penetration of fixative and embedding medium through the exoskeleton; also, the type of fixative and resin. This is for conventional LM and TEM. I would appreciate any suggestions. Thank you, Hank Adams EML New Mexico State University Las Cruces, NM email: hadams-at-nmsu.edu
We still do not know how it happened, because the listserver is not even in the address book of the computer from which the message was sent, but somehow a commercial message intended for a single person made its way to the microscopy listserver. We have to assume that somehow, somewhere we did something wrong. As a result, we are instituting a special program to make sure that it doesn't happen again.
Chuck Garber has asked me to send this message for him, because he has just left on an international trip.
Again, we're sorry.
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. 63 Unquowa Road Fairfield, CT 06430-5015 Ph: 1 203 254 0000 FAX: 1 203 254 2262 e-mail: AWBlackwoo-at-aol.com WWW--http://mail.cccbi.chester.pa.us/spi/spihome.html
The University of the West Indies in St. Augustine, Trinidad, is planning to offer a 2 week Workshop in Light Microscopy in the summer of 1996, subject to funding approval. The course is aimed at postgraduates, technicians, and faculty from all three campuses (in Trinidad, Jamaica, and Barbados), as well as participants from health, government and industrial organisations from the Carribean. I am looking for 2 - 3 teachers to help in the following areas: immunocytochemistry, karyotyping, and in situ hybridization, as well as the fundamental areas of imaging techniques and photomicrography. As far as possible it will be a hands-on course, bearing in mind that such equipment as a confocal microscope does not exist here. The purpose is more to present the capabilities of the techniques rather than exhaustively perform every method. There is tremendous need and interest in the Carribean for this kind of workshop. The funding agency will fund travel, accomodation, etc. I invite serious inquiries by FAX only (not e-mail, I am unsubscribing from this user group in self defense). Dr. G. F. Barclay, Plant Science Dept., U.W.I., FAX:809-645-7132
} To: psphicas-at-pipeline.com (Phil Sphicas) } From: gwe-at-biotech.ufl.edu (Greg Erdos) } Subject: Re: TEM-Carbon coated grids } } } } } Does anyone have a reliable way to make carbon-coated grids for high } } resolution work? } } I have tried floating carbon films off mica-- they seem to fall apart } } before I can pick them up on grids. } } Any tips or suggestions? } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } We routinely float carbon from mica and mount on sticky grids. } } after cleaving the mica, I cut it in 3-4 mm wide strips, evaporate the carbon by a resistance method (very important),. Then I clamp the strip carbon side up in one pair of tweezers, then I grab the other end and cut off a square of mica. I float it off in a white porcelain spot plate and pick it up on the grid, being careful to always blot from the side. } } Let me know how it goes } -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
Is there a reason why you cannot evaporate carbon on to collodion coated grids? The collodion quickly sublimes in the beam creating a very clear window to look through. I doubt that it would be much better with a floated film and the coating would probably be more beam stabile.
Dan
On Tue, 12 Dec 1995, Phil Sphicas wrote:
} Date: Tue, 12 Dec 1995 21:45:57 -0500 } From: Phil Sphicas {psphicas-at-pipeline.com} } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: TEM-Carbon coated grids } } } Does anyone have a reliable way to make carbon-coated grids for high } resolution work? } I have tried floating carbon films off mica-- they seem to fall apart } before I can pick them up on grids. } Any tips or suggestions? }
%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%% % % % Daniel Possin Work: 206/ 543-7489 % % Dept. of Ophthalmology RJ-10 FAX: 206/ 543-4414 % % University of Washington Home: 206/ 778-1714 % % Seattle WA 98195 USA Email: oemlab-at-u.washington.edu % % % % "The chinese expression 'cheung meng ba sui, gong hey fat choy' is % % equilvalent to Vulcan expression 'live long and prosper'. It's a % % small universe and getting smaller everyday". % % % %%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%%
} Does anyone have any suggestions for fixation and embedding small } crustaceans (clam shrimp) about 4mm in lenght?. I'm concerned about } penetration of fixative and embedding medium through the exoskeleton; also, } the type of fixative and resin. This is for conventional LM and TEM. } I would appreciate any suggestions. } Thank you, Hank Adams } EML } New Mexico State University } Las Cruces, NM } email: hadams-at-nmsu.edu
Hank,
Last July I asked a similar question and go the following answers. I have yet to try any of them so cannot add to them my experience. I'd appreciate any additional tips you get as I have not been able to find anything in the textbook literature.
Bob Wise
****************
Haven't had a whole lot of experience with insect tissue, but I have done some. If you get better advice from more experienced bug people great (but lest you get very little response). 1) Using a sharp razor blade, cut through the insect, exposing the interior to the fixative. Doing this in fixative has seemed to work well for the specimens I have dealt with. Cut through as close to the area of interest as possible. (For legs we found cutting the top and bottom of the leg was needed, I would assume the same would be true for things like antenna, etc.). Gills are penetrated by the fixative just fine (after all they are designed for chemical penetration). 2) Fixatives, insects tend to be hydrophobic and the addition of a wetting agent is needed. I have used both Tween and photo-flo with comparable results (just a few drops for say 20ml). 3) Use of a combination of 2-3% glut, 1% formaldehyde, and 1-3% (up to 10%) acrolein (acrylic aldehyde) works well. The acrolein is a recommended fixative for insects. I also recommended sodium cacodylate buffer. 4) En bloc staining with UAc helps with staining, and if you're interested specifically in walls barium permanganate post section staining, in addition to PbCit and UAc, does unbelievable things to the walls! 5) Physically breaking the exoskeleton allows penetration of the of the fixes, and even more lets the resins penetrate. I would use a hard mixture of a low viscosity resin, i.e. Spurr's or quetol 651 and extend infiltration times *****************
Microwave-accelerated chemical fixation methods have been used with success to fix insects for TEM:
1. Lindley VA: A new procedure for handling impervious biological specimens. Microsc Res Tech 1992, 21:355-360
2. Smid HM, Schooneveld H, Meerloo T: Microwave fixation of water-cooled insect tissues for immunohistochemistry. Histochem J 1990, 22:313-320
3. Contact me directly for detailed info.
Gary R. Login, D.M.D., D.M.Sc. Assistant Professor of Oral Pathology Beth Israel Hospital Department of Pathology 330 Brookline Avenue Boston, Massachusetts, 02215
Routine TEM fixation in our hands is 2.5% glutaraldehyde in 0.1M PO4 (with 0.15M sucrose) for 1-2 hours at 4 or 20 C; wash then post fix in 1% OsO4 for 1 hr in the same buffer; wash in dist water; dehydrated in a graded series of ethanol (acetone extracts lipids more) and infiltrate into your resin of choice.
If you have a time consuming dissection and/or don't want to breathe too much glut you should dissect under insect Ringers. Yes you will have trouble getting fix thru cuticle. At very least you should inject fix into body of larvae making sure it's in the haemolymph not the gut lumen. If insect is big (10mm) dissection is best (if possible). If not simply punch tiny holes thru the cuticle with needle / dissect off the terminal half mm(if not necessary for observation / cut larvae into 2 or 3 portions and fixseparately.
If you require more exact recipes contact us directly at: erich-at-ento.csiro.au
Eric Hines, EM Unit, CSIRO Entomology, PO Box 1700, Canberra 2601 Oz ************
I did some Drosophila scanning for Sean Carroll on campus; search for his lab's papers using SEM and confocal for possible technical ideas.
Mark W. Tengowski, DVM, MS, Postdoctoral Fellow, 309 Zoology Research, 1117 West Johnson Street, Madison, WI 53706 (608) 262-2048 lab, (608) 262-7319 fax ********
Dear Microscopists, I am in struggle with a possibly quiet simple and well understood problem. Upon observations on haevily deformed fcc and bcc metal powders, e.g. Ag, Cu, Fe and V, very typical electron-diffraction patterns were found, showing intensity modulations on the diffraction rings. This indicates that the materials are textured. In a book I have found a very similar diffraction pattern to those of Ag and Cu taken from gold leaf. Unfortunately the texture of the golg leaf is not specified in that book, but it appears to be a fibre texture. I would be happy if someone could answer to the following questions. 1. What is the texture of gold leaf and of the analogous treated bcc metalls, i.e. hammered Fe. 2. Where can I find references on these topics.
Thank you very much yours sincerely Christof Gente
Just off the cuff, I would say that one advantage of getting the PC interface would be that you could network the PC to your system and store images on your LAN, so that you wouldn't need a separate storage media. I have an even older Amray 1645 (1985 model) which is analog only, and therefore I have to use a scanner to produce images for transfer to customers electronically.
Let me know of the other responses to your questions, maybe something would be useful to me as well.
Thanks, Melanie Behrens Senior Chemist/Microscopist Texaco, Inc. Beacon, NY
Anyone have a good supplier for cellulose acetate sheets for making good resolution surface replicas of hard surfaces? Thanks.
############################################################################# John J. Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
The Alberta Microelectronic Centre is contemplating marketing 3mm TEM grids made from silicon, with each grid opening covered by a thin electron-transparent membrane of single-crystal silicon or of amorphous silicon dioxide or silicon nitride.
Possible applications might include direct TEM observation of thin films deposited on the membranes, or their use as a support for biological structures.
AMC would greatly appreciate advice from the TEM community on any of the following questions: (1) Would you (or someone you know) make use of such grids; if so, in what quantity ? (2) Are there other applications, not mentioned above ? (3) What is the ideal membrane thickness? Are there limits on the acceptable amount of variation in thickness? Is a grid-bar thickness of 0.4mm thickness compatible with most TEM specimen holders, or is 0.2 mm preferable ?
Please reply to me and I will forward the information.
Gentle networkers, Some time ago, I asked about the lifetimes in practise for 50 W mercury arc lamps. I recieved many answers, over an extended period. Many thanks, and here is a digest for anyone intested. The question also came up on the confocal list, so this summary is cross posted.
Robert Josephs (bob-at-befvax.uchicago.edu) wrote: I have a suggestion. Get a small fan (the kind used to cool electronic circuits) to blow air through the lamp housing. This keeps the temperature around 20-30 degrees instaed of the 200C or more. When the lamp runs this cool it last and lasts and.... I don't know how long they last now. Before I used the fan a 100W Hg lamp would last ca 400 hours but during the last 100 hours or so the intensity fluctiated a lot. Since I have been using the fan it has not been necessary to replace the lamp. The current lamp has 500 hours and is perfectly stable. *************
William Tivol {tivol-at-wadsworth.org} wrote: ... The biggest contribution to failure comes from thermal stress, so if the lamp is not turned on and off, but is stabile at a particular tempera- ture, it will last a long time. ...If you can arrange your work so that it can all get done in one stretch, you should not be afraid to try using the lamp for 400-500 hrs, but if you are using it in equipment which is turned on only when in use, change the lamp when the mfr says, unless you have a surplus of housings and a shortage of lamps. *************
Edward J. Huff (huffe-at-pgl7.chem.nyu.edu) wrote: We use HBO 100W/2 Zeiss lamphouse with (added) external fan, and run for 400 to 500 hours. Lamps are Osram HBO 100W/2. I understand that without the fan they burn out faster but I never ran them without the fan myself. *************
DALE A CALLAHAM {dac-at-oitunix.oit.umass.edu} wrote: A fan is a step in the right direction for keeping things cool, but usually isnt used since air currents on the lamp envelope can cause excessive gradients and stress (etc....kaboom!), though people may get away with it for the low power lamps. Also that isn't used in cases where photometric functions are required, for stability reasons. Osram suggests cooling of the lamp bases, the envelope needs to be hot to develop the designed operating pressures. ****************
shaun.sandow-at-anu.edu.au (Shaun Sandow) wrote: We run our Hg lamps (both 50 and 100W) for about 400 hours before replacing them. After this time the 50W lamps become dim and thus need replacing regardless of the manufacturers recommendation. *************
R.G.White-at-sci.monash.edu.au (Rosemary White) wrote: We've run our last two 50W HBO Hg lamps for 1) just under 300 h (a mistake! - changed when the fluorescence got too dim) and 2) just over 200 h. As long as they don't get turned on and off too frequently they seem OK. It gives our Leica rep kittens, though. ***************
Benjamin Walcott {bwalcott-at-ccmail.sunysb.edu} wrote: I find that I can run the 50 watt mercury arcs for about 170-190 hours before I find that they either begin to flicker or the intensity of the staining that I am using them to detect seems to drop in intensity. I have also been told, probably by an arc lamp salesperson, that if you run them too long beyond their normal life, they can explode. I have never had that happen in the 10 or more years that I have been using them. *******************
imgp-at-mbimp1.mbl.tno.nl (Kees van der Wulp) wrote: In our setup (an older Zeiss LSM), where we only use the 50W Hg arc lamp for focussing purposes in combination with a excitation filter that does not affect the fluorochrome of interest, I normally use it for 300 hours and then replace it. During the last 5 years an explosion only happened once. The reflective mirror was broken, but the collector in the lamp house was not damaged. You have to be careful because of the mercury vapour that is set free. I always have a spare refletor on stock (only $ 10 and a first componenent of the collector lens, just a few $ more).The savings by using the lamp 300 hours are far more than the price of the spare parts. I agree with William Tivol that switching the lamp on and off will surely shorten the life of the lamp dramatically. I was told by some reps that UV intensity drops remarkable after 100 hours of burning. I don't know wether that is true, but we don't use uv excitation. ********************
DALE A CALLAHAM {dac-at-oitunix.oit.umass.edu} wrote: The details of arc lamp operation are very simple, but also as complex as to the differing designs of equipment used for their operation.
To start with, the ignition process always damages the lamp at each ignition. How much damage depends on the ignitor design. The manufacturer's life suggestion is based on 30 min of operation per start using the brute force ignitor designs of earlier years. Some newer equipment is far more gentle at producing a stable initial discharge and this together with longer operation per start should be reflected in a longer life.
As for operation of the lamps, the electrode tips are eroded back to a wider separation with ignition and operation times. This larger spacing increases the voltage across the arc and if the lamp is operated in a constant current mode, this means more power dissipation in the lamp and thus more heat. Osram and others specify a maximum electrode temperature for which the lamps are designed. The design of the lamp holder and lamphouse will be important in determining the maximum power at which a lamp can be safely used.
Early arc lamp equipment and some of the simpler models today (AC models and the low priced unregulated models typically) use inductive ballasting to effectively operate at constant current, so users with this equipment should be cautious in exceeding recommendations.
Power supplies that operate in a constant power mode and ones that display power readings and are regularly adjusted are probably safer to use if playing with lamp life ratings, since the lamp temperature will be limited to the initial value and likelyhood of explosion is reduced. In the constant power mode, the current will be reduced as the electrode voltage incerases and the lamp will dim until the electronics can no longer maintain the arc (design dependent). I know that the HBO100W/2 can be operated at 1 amp with the right power supply.
I made some measurements on a number of 100W Hg lamps (HBO 100W/2) that I used and found that the initial arc voltage was in the range of 14.8V (warmed up and operating at 5 amps). The arc voltage at 200 hours was as high as 28V (-at- 5 amps) and this is 140 W!
A lamp blew the lab and trashed a quartz collector lens - price one of those lately? I would hate to see people lose good equipment for trying to squeeze out the last few hours of lamp life....it's not just an attempt by the vendors to sell more lamps, although I'm sure they don't mind. The lamps are rated safely, based on average characteristics.
Strech those dollars carefully! *************************
"Crossman, Harold" {crossman-at-rd.sylvania.com} wrote: My company manufactures 50W Hg microscopy lamps, and in reference to the recent postings I would like to offer the following suggestion. Like any other mechanical device, pushing a lamp beyond its designed capabilities can result in failure. I suggest changing lamps just like changing oil in a car; when scheduled, not when things break down. We have a national customer support center that can provide specific technical details. The toll-free number is 1-800-LIGHTBULB. *************************
ScottE57-at-aol.com (Scott E. Berman) wrote: I have previously worked for Zeiss for over 10 years in microscope sales - in that time I have found that the 100 DC HBO burners would run for 200 to 600 hours with the only problem of light output diminishing and problems then getting even illumination over the 175 to 225 hour mark. The 50W HBO AC is a different story - the bulb is rated at 100 hours but depending upon operating conditions they will cloud up and darken as fast as 60-75 hours of use and I have had in 15 years in the indusrty 4 blow up violently - they do not go quitely like the 100 DC models. All four were in the 75 to 150 hour range. As such I have highly encouraged people to change at 100 hours!! - the Bulbs are same price but upon intial purchase the DC power suplly is considerably more expensive - you pay up front and get long and safe bulb life or you save now and pay in the number of bulbs purchased. Depending upon manufacture the difference could be $300 for AC aupply vs. $2500 for DC supply. $2000 buys 10 to 15 bulbs and if you are a heavy fluoresence user than get the 100, if not the 50 is a good way to put more money into the optics where it belongs. ***************************
DALE A CALLAHAM {dac-at-bio.umass.edu} wrote: The question was about using arc lamps for more than the rated life. The characteristics of arc lamps are the key to understanding what may be reasonably done with them, but the wide variey of power supplies that are available are the other part of that equation and often little information is available to help users in this regard. Armed with the detailed characteristics of mercury arc lamps, please go after your manual or vendor for the missing part so that you may make the right decisions.
Mercury arc lamps are given a nominal lifetime rating based on 30 min of operation per start and operation at the specified conditions. Each ignition of a mercury arc does some damage to the electrodes. A sufficient current at a high voltage is required to break over the arc in the cold lamp where the mercury is not in the high pressure vapor state. This energy serves to vaporize some mercury and get the arc established, at which point it can be maintained at a lower current and voltage. This large surge erodes the tips of the electrodes, opening the spacing a little each time. Both in starting and in operation, the electrode spacing is involved in establishing the arc voltage at the operating current. When the spacing is too large, ignition and operation become more difficult as described below. The ignition circuitry that Osram, for example, describes in their literature of a few years back, is simple passive electronics that simply do the job. More modern electronic units CAN start the lamps more gently and this, together with longer operation per start, will be reflected in longer usable lamp life.
Mercury arc lamps have nominal ratings: a 100W lamp operated at the nominal 5 amps current will only give 100 watts at one brief part of its cycle. This is because of the change of the arc electrode spacing (and thus arc voltage). Most early power supplies, whether DC or AC used a choke to limit and stabilize the lamp current; since the arc voltage is much lower than the line voltage, this gave an approximation of constant current operation. I have found that a new HBO100W/2 lamp warmed-up and operated at 5 amps with a good DC supply has an arc voltage of about 14.8V, giving only about 75W. At 200 hours of life, this lamp had 28V across the arc (-at- 5 amps) and this is 140W! No doubt this relationship would continue until a) the lamp gets too hot and explodes, b) the power supply operating voltage becomes insufficient to maintain 5A current (giving a decrease in power and dimming of lamp output), or c) the power supply is unable to ignite and stabilize the lamp at the higher arc voltage. It is possible to operate the HBO100W/2 lamp at currents as low as 1 amp with the right equipment, although the light output is also way down. If you have a proper power supply and the light output is adequate, operating at less than the nominal rating should give a real extension of lamp life.
Another issue is the operating temperature of the lamp. Osram specifies that the lamp bases should be no higher than 230C degrees and that it SHOULD be operated at below 200C if possible. Lamp housings should be designed to provide sufficient heat dissipation at the nominal wattage by passive heat flow or air flow directed at the lamp bases: the quartz envelope must be hot to have stable Hg temperature and pressure but an excessive gradient along the lamp will produce excessive stress on the lamp. Clearly, with economy power supplies the power dissipation cannot be limited and at some point the lamp will get too hot and you risk catastrophic failure. More expensive electronic supplies allow for various sorts of automatic control or manual readout and adjustments to control or limit power levels.
In summary, there are very good reasons that modern equipment can give some lengthening of lamp life, but there are very real reasons not to push this too far: the lamps simply DO get consumed in the process of giving light. I have personally seen a 2" quartz collector shattered by a lamp explosion and that is quite an expense to replace. **************************************
Lots of people run them much longer than the specified time. There are potentially two big problems here:
1. After the life of the bulb is reached, the bulb starts a progressive drop in intensity. The first wavelength affected is the UV and it creeps up from there. Most people using FITC/Rhodamine, don't notice as much until the bulb is affected in their wavelength (400+ hours). If you are doing quantitative work, you will definitely want to change bulbs at the specified times.
2. Stability. The longer the bulb is run, the more the electrode and anode are worn. As they wear, the arc becomes unstable and dances on the ends. This causes flickering and some noise that can interfere with patch clamping. If a bulb explodes in your lamphouse, you will find that you now have to replace the collector lens, relay optics and sometimes the heat shields and mirrors. The biggest cause of exploding bulbs is on/off witching too frequently. A mercury bulb must be allowed to completely cool before be turned back on. A xenon bulb can be refired while still hot. To prolong the life of a bulb, it technically should be left on for long periods of time.
It should be noted that Osram has just come out with a new HBO 103 bulb which is the new replacement for the HBO 100W and is now rated to 300 hours for about $15-20 more than the old bulb. ***********************************
Robert Parsons {rparsons-at-INTERLOG.COM} wrote: HBO 50 AC Arc lamps have a rated lifespan 0f 100 hours according to Osram. (The manufacturer). This however varies according to the frequency of starting the burner.The fewer times you start the burner the longer the bulb life. The ratedtime of 100 hour refers to a average of two hours use for each start.
The rule of thumb is , If the bulb is used a lot, lots of starts change it every 100 hours. If the bulb is used for long periods, i.e. 4-8 hours at a time, you should get 150 hours.
If the bulbs starts to flicker after 100 hours change it, if you notice longing exposures change it.
For detailed information in the U.S. Contact Osram 1-800-431-9980 or in Canada 416-673-1996
In article "Stadden, David R" {DRSTADDE+aCENTRAL%Armstrong-at-MCIMAIL.COM} writes: } Date: Thu, 14 Dec 95 16:03 EST } From: "Stadden, David R" {DRSTADDE+aCENTRAL%Armstrong-at-MCIMAIL.COM} } Subject: Amray Image Archiving, etc.
. Is there something about our } particular configuration, i.e., Amray to PC system, that requires us } to go this comparatively expensive route to a dedicated computer with } hard drive?
NO. You can do what we have done and install an ImageSlave. These are inexpensive (about $3500-$5000 depending on resolution) boards which fit in a standard PC (386, 486, whatever you have) and digitise the photo signal from the microscope. Interfacing is very simple. We have 4. One on each STEM Unit and one on each SEM. Since they fit in a standard PC you can send the images out via ethernet, print them out on a laser etc. etc. Very versatile.
} Second, is there any way we could get more bang for the buck and } archive images from our PLM with common hardware?
We have installed a CD-ROM writer. These are now $1000 or less and CD blanks are around $10. Write two each time. A Gigabyte hard disk to store images until you have a CD full is about $300. CD readers are a virtual standard.
} } Apart from archiving, I'd be interested in hearing (especially from} Amray users of older scopes) about other image exploits. Sending them} to your customers? Feasible but can be slow and unreliable as everyone has climbed onto the net.
Printing them on laser printers? Of course. A 1200 DPI printer makes a great job of pix. The ImageSlave software runs in Windows so you can do instant prints off the microscope.
Photorealistic images are still pricey though.
Any special} considerations for future compatability?
Once you have images in the PC domain as a TIFF file you can do whatever you can imagine.} } Thanks for whatever you may have to share, } } Dave Stadden } DRStadde+aCENTRAL%ARMSTRONG-at-MCIMAIL.COM }
I am NOT a retailer, but this system has been very satisfactory for us.
In the USA, your contact is: Jim Hilton, Advanced Database Systems, 7931 South Broadway #322 LITTLETON CO 80122 Tel 303-761-5635
Mel Dickson, University of New South Wales. Sydney Australia.
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Andreas Brech Electron Microscopical Unit for Biological Sciences Department of Biology, University of Oslo. P.O.Box 1062 Blindern N-0316 Oslo 3 Norway Tel.: + 43-22 85 61 89 (work) + 43-22 43 83 23 (privat) Fax.: + 43-22 85 47 26 e-mail.: abrech-at-bio.uio.no
} Just off the cuff, I would say that one advantage of getting the PC interface } would be that you could network the PC to your system and store images on } your LAN, so that you wouldn't need a separate storage media. } I have an even older Amray 1645 (1985 model) which is analog only, and } therefore I have to use a scanner to produce images for transfer to customers } electronically. } } Let me know of the other responses to your questions, maybe something would } be useful to me as well. } } Thanks, } Melanie Behrens } Senior Chemist/Microscopist } Texaco, Inc. } Beacon, NY
Dear All, Melanie makes a good point about the advantages of having a PC connected to a network on your SEM, but where did you get the idea that only digital SEMs can have computer imaging on them? There are many PC systems for taking images off your SEM and storing them as digital images on the PC computer and as far as I know they all work fine on any SEM. There are both active systems, which have their own scan generators (beam steer) and passive systems which use the wave forms generated by your SEM's own scan generator to form the image on the computer (passive capture). Some of the systems I know of are: Quartz PCI, Image Slave (Australia), Printerface from GW, Semicaps and I'm sure there are others I have neglected. They will all interface to old, analog SEMs. I know of a Quartz PCI system running on a 1970 Nanolab and a Cambridge 250 (1980). The price of big hard drives is quite low now and the Iomega Zip drive, which stores 100 MBytes on each removeable disk is a reasonable price (~$225US). 1K by 1K by 8 bit images are less than 1MByte each in TIFF format, so you can get a lot on a gigabyte drive. Even burning your own CD-ROMs for image archiving has dropped to an almost reasonable price ($900US). Best of luck and have fun. You'll love your computer imaging system. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
We have directed this question to several researchers in the US and Europe, to the Usenet and now - as a last resort - to this microscopy list. Hopefully, I won't need to keep my fingers crossed for too long :)
We are trying to use SEM to study possible changes in morphology of spores of Bacillus licheniformes added to ice cream mix that contains 15% fat (which in turn undergoes a special treatment).
The first fixation protococol (glutaraldehyde & osmium tetroxide, centrifuging at all steps, etc) resulted in only being able to see the processed mix fat and no visible spore. It seems that a fine coat of lipid material (???) covers the "lumps", which we assume should be the spores. Acetone and chloroform was also used, but without any success.
We have/had no problem to visualize the spores when added to phosphate buffer; the SEM protocol used did work fine. However, it seems that the high fat content and the "fluid" nature of the mix are the real problem for us. Cheese, which is a solid matrix of fat + proteins, does not pose any problem. We can remove all the fat (leaving holes) and nicely see the bacterias present as well as the protein matrix (casein) that constitute cheeses in general.
Can anyone suggest a method of removing the fat and leaving the spores "uncoated" so they can be visualized during SEM preparation ?
Thanks for your comments and Merry Christmas to all.
Mary Mager mentioned PC-based systems (Quartz PCI, Image Slave, Printerface from GW, Semicaps). We have a Mac-based system to acuire digital SEM images from an older SEM. The interface is produced by 4pi, Inc.
Russell E. Cook Electron Microscopy Center for Materials Research Argonne National Laboratory 9700 South Cass Avenue MSD-212 Argonne, IL 60439 (708)252-7194 FAX: (708)252-4798
I need to embed plant material (leaf) for immunocytochemistry. The problem is that the protein we're looking for is believed to be embedded in the waxy cuticle. We're afraid of losing the protein with conventional dehydration and embedding protocols. I have no idea of the solubilities of the waxes which are esters if fatty acids and alcohols having 26 - 34 carbon atoms. Does anyone have any experience with this type of material and the use of water miscible resins. I know there are several different resins that are water miscible & commercially available, but are they applicable in this case and useful for immunocytochemistry? Thank you, Hank Adams EML, New Mexico State Univ.
Does anyone know of a good, reliable way to produce images of negatively stained specimens after immunogold labeling? I have tried the following two methods with bacterophages.
Method A. The protocol followed was: (1) Adsorb properly diluted phage sample onto glow-discharge plastic-carbon coated grid for one minute. (2) Incubate with 1st antibody for 30 minutes. (3) Rinse the grids with buffer, or pass the grids over drops of buffer. (4) Incubate with 2nd antibody-gold for 30 minutes. (5) Rinse again. (6) Negative stain. The difficulty with this method I have encountered is that after the procedure is applied either the film breaks as soon as the electron beam hits, or most of the phages are lost. (Without IEM, the negative-stained phages were fine).
Method B: Incubate the phages with 1st antibody and 2nd antibody-gold in a microfuge tube and then negative stain. This method works well, but the specimen appears to have a very high background of gold particles.
I wonder if others have similar difficulties and how they are dealing with them. Any information or suggestions would be appreciated.
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INSTRUMENTATION AVAILABLE: A Philips CM300 FEG equipped with a Gatan Imaging Filter and a Philips CM30 with a PEELS unit are the primary TEM instruments. There are a variety of associated instruments available including scanning electron microscopes, secondary ion microprobes, x-ray diffraction units, etc.
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} } Does anyone know of a good, reliable way to produce images of negatively } stained specimens after immunogold labeling? I have tried the following } two methods with bacterophages. } } Method A. The protocol followed was: } (1) Adsorb properly diluted phage sample onto glow-discharge plastic-carbon } coated grid for one minute. } (2) Incubate with 1st antibody for 30 minutes. } (3) Rinse the grids with buffer, or pass the grids over drops of buffer. } (4) Incubate with 2nd antibody-gold for 30 minutes. } (5) Rinse again. } (6) Negative stain. } The difficulty with this method I have encountered is that after the } procedure is applied either the film breaks as soon as the electron beam } hits, or most of the phages are lost. (Without IEM, the negative-stained } phages were fine). } } Method B: Incubate the phages with 1st antibody and 2nd antibody-gold in a } microfuge tube and then negative stain. This method works well, but the } specimen appears to have a very high background of gold particles. } } I wonder if others have similar difficulties and how they are dealing with } them. Any information or suggestions would be appreciated. } } Eleana Sphicas } Rockefeller University } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
In general I have always had success with your method #1. I would suggest a Blocking step after applying the phage and before applying the primary antibody.
Also use as small a mesh size for your grid as possible and use nickel not copper. I have always done it with 400 mesh grids, but smaller are available if you a re have trouble with the film breaking. Other wise just try to minimize mechanical damage as much as possible.
Good luck -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
} } } } We have directed this question to several researchers in the US } and Europe, to the Usenet and now - as a last resort - to this } microscopy list. Hopefully, I won't need to keep my fingers } crossed for too long :) } } We are trying to use SEM to study possible changes in morphology } of spores of Bacillus licheniformes added to ice cream mix that } contains 15% fat (which in turn undergoes a special treatment). } } The first fixation protococol (glutaraldehyde & osmium tetroxide, } centrifuging at all steps, etc) resulted in only being able to see } the processed mix fat and no visible spore. It seems that a fine } coat of lipid material (???) covers the "lumps", which we assume } should be the spores. Acetone and chloroform was also used, but } without any success. } } We have/had no problem to visualize the spores when added to phosphate } buffer; the SEM protocol used did work fine. However, it seems that } the high fat content and the "fluid" nature of the mix are the real } problem for us. Cheese, which is a solid matrix of fat + proteins, } does not pose any problem. We can remove all the fat (leaving holes) } and nicely see the bacterias present as well as the protein matrix } (casein) that constitute cheeses in general. } } Can anyone suggest a method of removing the fat and leaving the spores } "uncoated" so they can be visualized during SEM preparation ? } } Thanks for your comments and Merry Christmas to all. } } } } Sergio Feijoo } scf1-at-ra.msstate.edu } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } } }
The best thing you might do is to use cryo-SEM and fracture the frozen specimen, sublime away some of the ice, coat it and observe it in the frozen hdrated condition. This requires special aparattus. If you have a freeze-fracture device available it might be possible to do that and observe the C-Pt replicas either by SEM or better by TEM. If you don't have one, there is one at the Univ. of Georgia.
It might also be possible to dry fracture your sample in some way just before you coat it. I don't know what kind of a chunk you end up with or is it a powder. If it is a power, try putting it between two pieces of scoth tape and then rip it apart, hoping to expose your spores. If it is a piece, tease it apart with needles ar scalpels.
I have found high lipid stuff very difficult to deal with by conventional SEM -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
A colleague has asked for information. They are establishing a centrally administered SEM unit which will be available for researchers at his university. There will be a chargeback system and there will also be internal money or time available for instrument useage. This would be on a competitive basis (i.e., internal competitive money). Does anyone have in place a document or application that would detail procedures for applying for such internal research money or for scope time? Please direct your responses to Dr. Michael Dingerson, Assoc. Vice Chancellor for Research at mdingers-at-sunset.backbone.olemiss.edu OR you may phone him at: 601-232-7482. Many thanks and happy holidays to all! John
############################################################################# John J. Bozzola, Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
} MHS: Source date is: 19-Dec-95 07:55 EDT } } I am looking for a source of polarized light micrographs } (preferably of colourful materials ie LC, spherulites } etc.) that have been enlarged and are suitable for framing. } } Any suggestions? }
Try pp-DDT coming out of a melt. It give beautiful and colorful flowers. I use it regularly to give a "gift of flowers" to tours, watching them form on the video screen.
I hope this helps.
Shalom from Jerusalem, Azriel Gorski
++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Azriel Gorski, PhD Head, Optical Microscopy Laboratory Division of Identification and Forensic Science Israel National Police Jerusalem, ISRAEL
\We are searching an used SEM ( working, if it is possible )for received it in donation . Please, contact me for any questions.
E-mail : RNBALDUC-at-arcide.edu.ar
Fernando Balducci Laboratory of Electron Microscopy School of Bioengineering. National University of Entre Rios. C.C 57 Suc 3 Parana - Entre Rios Argentina.
I have had several questions about what I mean when I refer to resistance evaporation. If current passes directly through the carbon source and thus heating it, this is resistance evaporation. So if one is using carbon thread or a pair of carbon rods that touch, you are using electrical resistance to generate the heat.
The alternate method is electron beam evaporation where the carbon source is positioned in the center of a tungsten coil, but not touching it. When current is applied to the tungsten under vacuum, electrons are produced which then bombard the carbon and thus evaporate it.
Sorry for the confusion. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
We have been using LR White embedding medium for LM and EM immunocytochemistry in our lab for several years now and lately we have been having a problem we have been unable to solve. LR White has always been more difficult to mount to slides and/or grids than other mediums, but the problem has seemed to become worse. It has become particularly annoying to find that it has developed many small puckers or bubbles in sections mounted on formvar coated grids after processing for immunogold labeling. We believe that there may have been a change in the formulation of the product because sections from blocks embedded over a year ago do not seem to have this problem and our embedding procedure hasn't changed. Are any of you having this same problem? Have any of you solved this problem? If you don't have this problem, please tell me what you are doing so we can compare notes. Thanks in advance.
% Daniel Possin Work: 206/ 543-7489 % % Dept. of Ophthalmology RJ-10 FAX: 206/ 543-4414 % % University of Washington Home: 206/ 778-1714 % % Seattle WA 98195 USA Email: oemlab-at-u.washington.edu % % % % "The chinese expression 'cheung meng ba sui, gong hey fat choy' is % % equilvalent to Vulcan expression 'live long and prosper'. It's a % % small universe and getting smaller everyday". %
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
I have seen the "commerical" advertising posted by Evex Analytical. Since the postings continued after a warning from me. I have removed their address from the server.
Thank you for expressing interest in the VIDX Scan Active Digital Imaging System, the Fast, High Resolution SEM Image Acquisition.
Features VIDX Scan Active Digital Imaging System, - Menu Driven - Quick button operation - X-ray mapping in minutes - Multisouces acquisition - 12 Bit high resolution image acquisition 4096 gray Levels - Independent contrast for each source - User defined resolution - Pixel-by-pixel, multi-signal digitization
Now capture up to 6 high resolution images from SEM, STEM, EDS, WDS. Images are scanned from the SEM, directly into the PC as an image file. All images are stored in standard TIFF format. Archive your images on your Hard Drive, Tape Backup, CD Writer. Process your images using Image Pro, the world's greatest image processing software. Write Reports - Import images effortlessly into your word processor. Print to the 1200x600 Laser Printer giving Photo realistic quality. Network your system via the PC.
You will be able to capture images quickly and economically without the use of expensive and time consuming micrographs.
The VIDX Scan Active Digital Imaging System, actively controls the SEM Scan, thus allowing multi-frame image integration. Multiple time constants allow the scan to be slowed for high noise situation such as low voltage or low spot size conditions. Images can be stored with SEM data (micron bars, user comments) permanently overlaid on the image, or stored in the image file without overwriting any part of the image.
The pixel-by-pixel, multi-signal digitization method will assure the precision necessary to overlay images acquired from different sources.
If you would like to learn more about VIDX Technology products contact your Service & Sales representative at (908)874-3800. We look forward to satisfying your X-ray Microanalysis & Imaging needs.
[Please, skip this message if you like, and I am sorry for filling up your mailbox. I tried several times to reply directly to the author with out success]
Well, me throw in my two cents worth:
} We're looking to begin electronically archiving our SEM images from } a vintage 1988 Amray 1820. (This is digital imaging, but not PC } controlled.)
I take it by "digital imaging, but not PC controlled, what you mean is that you already have a bunch of digital images, but they're in an Amray format and you are wishing to store them on a standard PC format media, in a standard format (TIFF,PCX, GIF, etc.) yes?
Assuming I've got this correct than you have just two problems: (1) Getting the Armay stored images onto a PC based system. This is done either by getting a some type of drive which will read the Amray disks and then re-save them to a PC-format disk drive, or "networking" the Amray system to a PC. (Let me address the second problem first and come back to this one below) (2) You have image files in an Amray format, which will need to be comverted into a standard image format (just about all PC based imaging software will handle a wide number of image formats, and will easily handle conversions between standard formats, which format you choose is of little consideration in this situation BUT should be considered since some formats will result in a reduction of data, i.e TIFF of an image is not equal to JPEG of the same image). But unless you know that the Amray images are stored in a standard format (i.e. TIFF most likely) or someelse out there can provide you with and Amray to something else conversion program, you will either have to write your own conversion software or have to get this conversion software from Amray.
Networking the Amray with a PC: This can be done a number of different ways: (1) via an ethernet connection, (2) via a serial line connection, or (3) via a high speed SCSI connection. And aparently Amray suggests the SCSI connection. This is a very common PC (and Mac.) interface, which means its commonly available and realatively inexpensive. SCSI (pronounced Skuzzy) currently has three PC configurations: SCSI - I, SCSI - II (Fast) , and SCSI-III (Fast & Wide). I would guess that the Amray will handle SCSI - I, but they are downwardly compatible, i.e. SCSI-III will handle -II & -I. Not having talked with Amray, or having experience with this Amray situation, I don't know exactly how they plan on handling the data transfer but my guess is basically the Amray system (Microscope) will be treated as simply a "device" (Like a printer, scanner or external diskdrive) connected to the PC system via the SCSI. And you will be able to copy data from the Amray system directly to the PC system and store it to disk if you want. You can add SCSI to any 386 or higher PC system (386 will only handle SCSI-I, a 486 or pentium is needed for SCSI-II or -III). And I strongly suggest you get an Adaptec SCSI controller (Prices range from ~$100 - $300) as I have found them to be very reliable and very strongly support throughout the industry. What you can do is purchase your own PC computer system, to handle whatever you would like so long as it can also handle SCSI (~$800 for a bare bones system, upto ~6,000 for a really great imaging system, but you can figgure on ~$2,000-2,500 for a good solid system (I do not recommend a notebook or laptop system for this purpose), and have it run DOS/Windows, OS/2, Windows 95 or Windows NT (UNIX can be a pain in the .... neck?) whatever your pleasure. And feel free to make it networkable.
SCSI drives: Each SCSI controller card can handle upto 7 devices connected to it (SCSI-III upto 15). Therefore you can use the SCSI controller to connect (1) the PC hard disk, (2) the AMRAY connection, (3) a removable storage device, and still have room for 5 more devices (i.e. a CD-ROM drive and/or a scanner)
} About the actual mechanics of doing this, I have a couple } questions. First, Amray recommends a system they will supply, that } consists of a dedicated computer with normally big hard drive, } monitor, and image transfer utility via SCSI interface. You get } about 3 TIFF images/megabyte of hard disk storage. I'm not well } versed in this field, but seem to recall hearing more than once } about inexpensive magneto-optical drives and cheap, removable data } storage disks that hold lots of images. We are PC, not Mac based, } and are corporately networked. Is there something about our } particular configuration, i.e., Amray to PC system, that requires us } to go this comparatively expensive route to a dedicated computer } with hard drive? Second, is there any way we could get more bang for } the buck and archive images from our PLM with common hardware?
Removable drives: There are a number of options here.
(1) Magneto Optical disk drives, these can store upto 4.6Gb. Common storage sizes are 650Mb, 1.3Gb, and 4.6Gb $800 for a 650Mb (Panasonic), $999 for a 1.3Gb (NEC), $1700 for a 1.3Gb ( SONY or Panasonic) to $2100- 2,600 for a 4.6 Gb drive (Prices vary on disk size and speed for writting and reading). Disk prices vary from $65/650Mb, $90/1.3Gb, and 4.6Gb/ ? ~$135. These drives are the fastest, nearly as fast as current hard drives, thought they write slower than the read. Generally the larger drives can handle the smaller disks, but this may vary depending on price for the drive (more $$ for more flexiblity). These disk cartridges are 1cm high by 15cm wide. The drives are single sided, i.e you have to remove, flip and reinsert the disk. But they are 'infinitely' rewrittable (some are WORM - write once read many), and they are randomly readable. I have worked with these for three years now and haven't had many problems with them.
(2) remoavble hard disks: these can store from 44Mb (Syquest or Bournolli), to 270Mb. Common storage sizes are 44Mb, 88Mb, 105Mb, 135Mb, 200Mb, and 270Mb. And the dirve prices vary from $239 to $600. Disk prices vary: $50 / 44 Mb, $65/105Mb, $70 /200Mb. These are fast, the disks don't hold much and you don't really save much $ per disk, but they are more common to find others in your area with the ability to read/ write with disks.
(3) CD-ROM recordables: These are strickly write-once read many (WORM). Drive prices have fallen to ~$1000 (2x or 4x) to $3000 for 6x. The disk cost $10, and hold upto 650Mb. Adavantage: cheaper media, VERY common reader drives, very stable for LONG term storage. Disadvantage: problems writting disks (you would still have to copy your data to a harddisk and ten write them to the CD-r, very slow writting disks, slow reading disks, WORM.
(4) Zip drives: very cost effective, but very slow since the data is compressed both during writting and reading.
(5) Tape drives: Too slow for anything but long term back up storage.
} Apart from archiving, I'd be interested in hearing } (especially from Amray users of older scopes) about other image } exploits. Sending them to your customers? Printing them on laser } printers? Any special considerations for future compatability? } Thanks for whatever you may have to share, Dave Stadden } DRStadde+aCENTRAL%ARMSTRONG-at-MCIMAIL.COM
There I think I have written far too much for you to consider at present, so I'm not going to comment on anything else at the moment. But do feel free to ask if anything I have written doesn't make sense. I'm in the never ending process of developing a digital imaging network here in my facility, which is why I have all the above info in hand.
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
Anyone having difficulty reaching the organizers of this conference using the telephone and fax numbers quoted, for example, in the Proceedings of the RMS, should note that there should be an extra "2" in the numbers.
Dr E.C. Chew's correct telephone and fax numbers are:
tel: +852 2609 6853 fax: +852 2603 5031
Robin Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za (tel: +27 461 318168 - fax: +27 461 24377)
Have you looked at Tomer's book Structure of metals through optical microscopy? The english edition is from American Society for Metals, Metals Park Ohio, but if I recall correctly the original hebrew edition has many more nice pictures. The ASM book quotes his location as Nuclear Research Center Negev, Beer Sheva, Israel. Prof.Walter A.Mannheimer Metallurgy and Materials Engineering Federal University of Rio de Janeiro POBox 68505 21945 Rio de Janeiro Brazil Voice 5521 280-7443 (Dept.office) 5521 590-0579 (direct) Fax 5521 290-6626 Email: wamann-at-metalmat.ufrj.br
Does anyone have any hints on reducing the background and debris from DAB labelling?
Ted Bell, bell-at-medcolpa.edu Medical College of Pennsylvania and Hahnemann University Neurobiology and Anatomy http://ussanatomy.medcolpa.edu/FaberWeb/confocal.html
} } Does anyone know of a good, reliable way to produce images of negatively } stained specimens after immunogold labeling? I have tried the following } two methods with bacterophages. } } Method A. The protocol followed was: } (1) Adsorb properly diluted phage sample onto glow-discharge plastic-carbon } coated grid for one minute. } (2) Incubate with 1st antibody for 30 minutes. } (3) Rinse the grids with buffer, or pass the grids over drops of buffer. } (4) Incubate with 2nd antibody-gold for 30 minutes. } (5) Rinse again. } (6) Negative stain. } The difficulty with this method I have encountered is that after the } procedure is applied either the film breaks as soon as the electron beam } hits, or most of the phages are lost. (Without IEM, the negative-stained } phages were fine). } } Method B: Incubate the phages with 1st antibody and 2nd antibody-gold in a } microfuge tube and then negative stain. This method works well, but the } specimen appears to have a very high background of gold particles. } } I wonder if others have similar difficulties and how they are dealing with } them. Any information or suggestions would be appreciated. } } Eleana Sphicas } Rockefeller University
I have had sucess in the past with a slight variation of method B:
My antibody-labeled sample was incubated with Protein A-gold for 60 = min at 20=B0C, and then separated from the unbound labed by wasing (four cycles) in phosphate-buffered saline, employing a Beckman airfuge centrifugation conditions of 134,000g with rotor 95A or at 196,000g in the 110A rotor.
The final pellet was taken up in 10=B5l of buffer and then negativel= y stained for electron microscopy.
Message-Id: {199512211718.LAA21069-at-Sparc5.Microscopy.Com} Comments: Authenticated sender is {semlab-at-mail.ims.uconn.edu}
Hi! I'm a Polymer Science student, and I would like to know which LM techniques are used to prevent evaporation of volatile solvents from solutions. Special slides & cover slips? Special sealing waxes? Thin cuvettes with Teflon stoppers? Other ideas? Solvent example: heptane.
I'd greatly appreciate your suggestions! Happy Chanukah and Merry Christmas to you all!
Sincerely, UConn/IMS SEM Lab (from David J. Moonay)
We have recently experienced some problems with frozen tissue samples being damaged at some point before they are sectioned. We are looking at teeth that have been fixed in aldehydes then decalcified in formic acid/formate before freezing and sectioning. I suspect that the culprit is either the freezing procedure or the storage of the frozen blocks. The tissue that has disappointed us was frozen in OCT slowly in the cryostat and then stored there. I realize that for unfixed tissue 'snap' freezing is usually advocated but thought that gradual freezing was kinder if the tissue had been stabilized by chemical fixation. I realize this is a nieve point but we may be missing something obvious. I would appreciate hearing what other people use even if it is mundane and routine.
Ted, How about providing some specifics about your immunolabelling protocol and sample type? There are a number of ways to get background.
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Thu, 21 Dec 1995, Ted Bell wrote:
} Does anyone have any hints on reducing the background and debris from DAB } labelling? } } } Ted Bell, bell-at-medcolpa.edu } Medical College of Pennsylvania and Hahnemann University } Neurobiology and Anatomy } http://ussanatomy.medcolpa.edu/FaberWeb/confocal.html } } }
A newly created web page for activities of the Fermin laboratory at Tulane Department of Pathology is now available: http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html Send your feed back. Thanks, Cesar
What is the damage and which type of microscopy are you doing? The problem might be the formic acid solution, very very rough on tissues. Is the calcium all removed?
Best of luck Ed Calomeni Medical College of Ohio Toledo, OH emlab-at-opus.mco.edu
Subject: Time: 3:07 PM OFFICE MEMO RE: Texture of Au Leaf Date: 12/21/95
The book "Theory and Practice of Electron Diffraction" by Thomson and Cjochrane, Macmillan, 1939 contains a considerable amount of discussion on electron diffraction patterns obtained from gold leaf. W. C. Bigelow (bigelow-at-umich.edu)
Last week I posted a request for information about a new style of loop for the collection of ultrathin sections. Many thanks to all who responded. I was particularly impressed when I received a phone call from one vendor who recognized that I was decribing a competitor's product and phoned with the information anyway. Below is a summary of the information I received.
Many of you reported that I was recalling the "Perfect Loop (TM)" from Electron Microscopy Science. (I think I'm OK in mentioning the trade name, I have no commercial interest in EMS and, by the way, I have deleted price info from the summaries below.) Others suggested using slot grids. We will try both approaches and see which is best...
Jean-Louis Lavergne wrote: } I'm using a perfect loop from Electron Microscopy Science. It size fits } perfectly with the grid diameter. I'm not sure that this "concentrate" } the sections to the center but the meniscus formed during the picking of } the section is helpful for "destressing" room temperature sections. } } This instrument is really nice to have if you have to collect Room temp } sections. For cryo work I've never try it.
Scott Walck {walcksd-at-ml.wpafb.af.mil} wrote: } Electron Microscopy Sciences sells them. Talk to Stacie Kirsch. I have one } and they work well for what I do. I collect films that I float off NaCl } crystals. If you like, I can let our microtomist try it out and she can get } back to you
John. J. Bozzola {bozzola-at-siu.edu} wrote: } Could you be referring to the "Perfect Loop" made by Electron Microscopy } Sciences (215-646-1566). [price info deleted] I have used one and they } work fairly well on uncoated grids. They're almost perfect but we usually } pick up grids onto slots and then deposit the grids with sections floating } on the retained droplet onto Butvar films.
Mike Veith {veith-at-wustlb.wustl.edu} wrote: } ... I use the EM version and never section without } it. It's the next best thing to sliced bread--in my opinion! It does what } the vendor says it will.
And thanks for similar information from rw9-at-psu.edu (Rosemary Walsh) and Patty Jansma {plj-at-manduca.neurobio.arizona.edu} for info similar to the above.
Paula Falk (pmf-at-avery.med.virginia.edu) wrote: } I did not see the advertised product. I have always used slot } grids with on corner folded up for easier pickup. The slots } concentrate the sections in the middle on mesh grids, and with } the aid of an eyelash they can be aligned to the filmed slot grid } so all sections are on the slot. This also prevents stress on } filmed slot grids because they don't have to be submerged in the } boat.
And Chris Pella also wrote to suggest slot grids, passing on K. Chien et al., "A simple procedure for obtaining clean sections for TEM," 42nd EMSA Meeting, pp. 42 (1984).
Rex, Snap freezing will give better results even for fixed tissue. Fixation usually will further improve morphologic detail over that obtained with unfixed samples because of greater resistance to crystalization of fwater. Slow freezing allows formation of more and larger ice crystals within the tissue. For example, slow freezing of vibratome sections helps antibody penetration by ripping big holes in the membranes. Although formic acid solutions are better than other acids for preservation of detail, you might experiment with warm EDTA for better results. Further discussion of decalicification techniques may be found in "Preparation of Decalcified Sections" by Edward B. Brain, pub. by Charles C. Thomas.
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Thu, 21 Dec 1995, Rex Holland wrote:
} We have recently experienced some problems with frozen tissue samples being } damaged at some point before they are sectioned. We are looking at teeth } that have been fixed in aldehydes then decalcified in formic acid/formate } before freezing and sectioning. I suspect that the culprit is either the } freezing procedure or the storage of the frozen blocks. The tissue that has } disappointed us was frozen in OCT slowly in the cryostat and then stored } there. I realize that for unfixed tissue 'snap' freezing is usually } advocated but thought that gradual freezing was kinder if the tissue had } been stabilized by chemical fixation. I realize this is a nieve point but we } may be missing something obvious. I would appreciate hearing what other } people use even if it is mundane and routine. } } Rex Holland } }
Hi We usually use laser diffraction to determine the size distribution of our product, which is a suspension of roughly 15 micron diameter (range 5 to 50 or more microns) translucent spheres in saline. I want to use light microscopy in order to verify that instrument's accuracy. But it seems that the errors in microscopy are almost as large as the error of the laser diffraction instrument. Has anyone found a good solution to this?
My problem is basically that I want to measure particles that have a range of diameters, and that have settled on to the surface of the glass slide, so the equators are all at different heights above the slide. I've been using the 40x objective in brightfield mode with the condenser aperture stopped down to keep the depth of field greatest, but just now realized what that (N.A. 0.2) does to the resolution. Guess I'll have to take a huge number of pictures with the 100x oil objective & aperture open. Still, if I have several particles in each field, on average it will be rare to have the equator in focus. I'd rather not spend time working this out if it has already been done. Any suggestions?
The translucent particles act as lenses, so the apparent diameter is usually larger than the actual diameter, and I'm hoping to be very accurate. For instance, Duke latex beads 15.03 microns diameter (R.I. ~1.5) appear 16 microns if I measure individual particles, though a chain of 10 beads (to minimize error per bead) gives 15.1 microns per bead. And this is with the particles all in focus ! (The high R.I. seems to exacerbate the problem.) The apparent diameter seems to vary with refractive index, radius of curvature, and whether the plane of focus is above, at, or below the particle's equator. I'm interested not only in mean size, but also in the shape of the distribution out in the tails. Any suggestions? Have I reached the limits of the technique? For that matter, can anyone tell me how to calculate how many particles I need to measure in order to get acceptible statistical error in a given size interval out in the tail of the size distribution?
Please respond directly to me. Thanks! Richard_Thrift-at-Depotech.com
Although I've inquired on this subject before, things change, and I'm wondering what may be new. We're looking to begin electronically archiving our SEM images from a vintage 1988 Amray 1820. (This is...
Although I am not an expert in the theory and methods of freezing biological tissues, I do know that the techniques used to prevent ice crystal formation are not trivial.
Vitrification (freezing without ice crystal formation), although theoretically simple, is not easy to achieve in practice if cryoprotectants are not used. The theory is to remove the heat so fast that the water has no time to form a crystal. To remove the heat this fast requires special equipment and very small specimens. Snap freezing, which I assume means plunging a specimen into a beaker of liquid nitrogen or pentane, is not usually sufficient to freeze large histological specimens without ice crystal damage. Neither will fixing the specimen prevent ice crystal damage.
The simplest solution for preventing freezing damage in routine histological specimens, which are to be sectioned in a cryostat, is probably to cryoprotect the fixed material before plunging in liquid nitrogen. Tokuyasu has shown that sucrose solutions that are over 1.6 M can be frozen by immersion in liquid nitrogen without ice crystal damage.
As an aside, I thought that the discussion about being able to quantify the number and size of ice crystals in frozen tissue was effectively closed in the 1980's by the work of Dubochet, Unwin and others.
My advice to any histologist worried about the enormous tissue damage that occurs when freezing fixed or unfixed specimens in cryogenic liquid, is to cryoprotect before freezing, then freeze by immersion in liquid nitrogen. Sucrose will easily penetrate fixed tissues but should not be used as a cryoprotectant for unfixed material.
We are interesting by a camera equipment for HITACHI H9000 NAR in order to perform High resolution in solid state chemistry area. Could you send me your user opinion about TV rate intensified or Slow Scan camera such GATAN or LHESA and so on ( Resolution, FFT convenience, performance...). TV rate: GATAN 622, 692, 672 LHESA LH72LL SLOW scan: LHESA GATAN Thank you Sincerely your,
} Can anyone recommend introductory texts (theory, application and technique) } for confocal microscopy and cryomicrotomy ?
Regarding Confocal microscopy I recommend the second edition of the *Handbook of biological confocal microscopy* (Editor: James Pawley; Plenum Press, N.Y., 1995). Probably far beyond an introductory text, it is a really comprehensive collection of all topics regarding confocal microscopy.
-Dietmar-
+++ Dietmar Reiter {dietmar.reiter-at-uibk.ac.at} ++++++++++++++++ +++ Dept. of Zoology and Limnology, University of Innsbruck ++++ +++ Technikerstrasse 25, A - 6020 Innsbruck, Austria +++++++++ +++ ph: (+43)-512-507-6170 (-6161), fax: ..-507-2930 (-2957) +++
Does anyone know of a formal or informal users group for either the Visilog image analysis program (under Windows in particular) and/or the Link ISIS EDS system?
I am beginning some image analysis in earnest under the Visilog system on a Link and have run into a number of difficulties. Things aren't always working as I think they ought. While I think know image analysis, I definitely do not know its implementation on this software. Maybe I am misunderstanding the documentation.
I am also particularly interested in developing a custom script or scripts so that we can have a fairly automatic system. Anyone out there with experience in this area? ---------------------------------------------------- Warren E. Straszheim 270 MD Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091 E-Mail: wes-at-ameslab.gov (or: wesaia-at-iastate.edu)
coal characterization and processing electron microscopy, x-ray analysis, image analysis computer applications
As Dietmar says, Dr. Pawley's book is an excellent reference. For a more practical approach, see Methods in Cell Biology, Vol. 38, edited by Brian Matsumoto, and for theory, Confocal Microscopy, edited by T. Wilson (Academic press) is a good choice.
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
On Thu, 21 Dec 1995, Fred Hayes wrote:
} Can anyone recommend introductory texts (theory, application and technique) } for confocal microscopy and cryomicrotomy ? } } Thank you, } } Fred A. Hayes } 916-678-6280 } }
THANKS to all who responded to my queries about use of Si-membrane grids. I will discuss the results with AMC and provide individual answers (in January) to some of the questions raised. Ray Egerton.
Message-Id: {199512221930.OAA29935-at-thomas.ge.com} X-Authentication-Warning: thomas.ge.com: daemon owned process doing -bs
Merry Christmas
Please sign me up to this newsgroup.
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
Dear Richard, Do you have the possibility of checking your diffraction and LM particle size distributions with measurements taken from either digitized SEM photomicrographs or from SEM images collected and measured with image analysis software? Scored sample holders or finder grids are aids to insure the scanning of the area covered by sample particles. I generally use this method when checking on volumetric distributions. I would fix,dehydrate, critical point dry and sputter-coat "wet" spheres for examination in a conventional SEM. Other possibilities include the use of a CRYO-SEM or a Wet SEM. Rosemary
At 7:05 PM 12/21/95 -0800, Richard Thrift wrote: } Hi } We usually use laser diffraction to determine the size distribution of our } product, which is a suspension of roughly 15 micron diameter (range 5 } to 50 or more microns) translucent spheres in saline. I want to use light } microscopy in order to verify that instrument's accuracy. But it seems } that the errors in microscopy are almost as large as the error of the laser } diffraction instrument. Has anyone found a good solution to this? } } My problem is basically that I want to measure particles that have a } range of diameters, and that have settled on to the surface of the glass } slide, so the equators are all at different heights above the slide. I've } been using the 40x objective in brightfield mode with the condenser } aperture stopped down to keep the depth of field greatest, but just now } realized what that (N.A. 0.2) does to the resolution. Guess I'll have to } take a huge number of pictures with the 100x oil objective & aperture } open. Still, if I have several particles in each field, on average it will be } rare to have the equator in focus. I'd rather not spend time working this } out if it has already been done. Any suggestions? } } The translucent particles act as lenses, so the apparent diameter is } usually larger than the actual diameter, and I'm hoping to be very } accurate. For instance, Duke latex beads 15.03 microns diameter (R.I. } ~1.5) appear 16 microns if I measure individual particles, though a chain } of 10 beads (to minimize error per bead) gives 15.1 microns per bead. } And this is with the particles all in focus ! (The high R.I. seems to } exacerbate the problem.) The apparent diameter seems to vary with } refractive index, radius of curvature, and whether the plane of focus is } above, at, or below the particle's equator. I'm interested not only in } mean size, but also in the shape of the distribution out in the tails. Any } suggestions? Have I reached the limits of the technique? For that } matter, can anyone tell me how to calculate how many particles I need to } measure in order to get acceptible statistical error in a given size interval } out in the tail of the size distribution? } } Please respond directly to me. } Thanks! } Richard_Thrift-at-Depotech.com
_________________________________________________________________________ Marija Gajdardziska - Josifovska AssistantProfessor Department of Physics University of Wisonsin Milwaukee P.O.Box 413 Milwaukee, WI 53201 phone: (414) 229 4965 Fax: (414) 229 5589
Subject: Time: 4:43 PM OFFICE MEMO RE} Texts for confocal and RE} Texts... Date: 12/22/95
The question about what is a useful introductory text for cryomicrotomy has two answers. The first is if the cryomicrotomy is to be performed on unfixed, vitrified material. A useful introductory text can be found in the Royal Microscopy Society Handbook series (number 21, I think). The authors are Roos and Morgan and they thoroughly cover the basic theory and practical details of rapid freezing, vitrification and sectioning.
There are also two chapters covering the second subject, cryomicrotomy of fixed, cryoprotected material for immunocytochemistry. However, the best reference work on immunocytochemical methods is still the Griffiths book (1993, Springer Verlag, "Fine Structure Immunocytochemistry").
An even better way of picking up this technique is to attend a course (or visit a laboratory) specializing in cryosectioning.
Good Luck and Happy Holidays. Paul Webster, Ph.D. Center for Cell Imaging Yale University School of Medicine.
We import microscopes from China. They are not used, but high quality at pretty low prices.
At this time, we have quite a few stereos, including single power, dual power, and zooms. We have a few demo models with some discount at this time of year.
Please contact us if your colleague has any interest in more information.
Arthur Gillman Argyle International Inc Princeton, NJ
We import microscopes from China. They are not used, but high quality at pretty low prices.
At this time, we have quite a few stereos, including single power, dual power, and zooms. We have a few demo models with some discount at this time of year.
Please contact us if your colleague has any interest in more information.
Arthur Gillman Argyle International Inc Princeton, NJ
I was at a government sale and in the process of bidding on 'lots' of goods I aquired an Hitachi HU-12 electron microscope. It is super heavy, but I managed to get it into my shop. It apears to have tipped over at some point in it;s life and a few adjustment knobs are bent/broken. What I would like to find is documentation on it. I went through a telephone marathon only to find a guy that had it but would not help? Can anyone out there give me any assistance. I would like to make the thing operational as a diversion from .... Thanksin advance for any help or clues to find help. Don Hall
Mr-Received: by mta RANDD; Relayed; Tue, 26 Dec 1995 10:24:40 -0600 Mr-Received: by mta MCM$RAND; Relayed; Tue, 26 Dec 1995 10:24:41 -0600 Mr-Received: by mta RANDB; Relayed; Tue, 26 Dec 1995 10:24:53 -0600 Alternate-Recipient: prohibited Disclose-Recipients: prohibited Content-Return: prohibited
30% sucrose in PBS is a good cryoprotectant for lungs and would probably work for hard tissues, also. Fix the tissues first, then remove fixative in three changes of PBS (10 minutes each for lungs, probably longer for your tissue). Soak tissues in cryoprotectant at least overnight, at 4C. Blot off excess liquid, then snap-freeze the tissues in embedding medium. I found that tissues embedded in Lipshaw's medium were easier to section than those embedded in OCT. You might give the Lipshaw medium a try. I also concur with the comment about your decal solution - I think EDTA is considered to be a "kinder and gentler" decalcifying agent than acid treatments.
Good luck!
Jane A. Fagerland, Ph.D. Dept. Cellular and Microscopic Research Abbott Laboratories Abbott Park IL 60064