I need to make samples disbersed on carbon-coated grids for collecting some EELS standards of various oxides in powder form. I have Maganese, Titiatium, and Ruthenium oxides in various states. Anyone know a good prep that will evenly disburse a thin layer onto a carbon-coated grid? My first thought would be to mix with a solvent and put a drop on a grid, but I figured someone here has probably already done this.
Thanks in advance, Leslie
Leslie Krupp (Thompson) IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
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On Jun 1, 2006, at 8:40 AM, lkrupp-at-us.ibm.com wrote:
} I need to make samples disbersed on carbon-coated grids for collecting } some } EELS standards of various oxides in powder form. I have Maganese, } Titiatium, and Ruthenium oxides in various states. Anyone know a good } prep } that will evenly disburse a thin layer onto a carbon-coated grid? My } first } thought would be to mix with a solvent and put a drop on a grid, but I } figured someone here has probably already done this. } Dear Leslie, My choice would be your first thought--it worked very well for river-bottom sediment. I would try various dilutions to find the one that gives the desired distribution of particles, and I would glow-discharge the grids. If using water leads to aggregation, try an organic solvent, and if the particles still have a tendency to aggregate, try putting the grid with the drop of solvent in an oven--faster evaporation might overcome that tendency. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
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Title-Subject: [Filtered] Equipment for phase extraction from steels
Question: Dear All,
I need to choose equipment for electrolytical phase extraction from steels (carbides, nitrides, etc.) and I need any advice about equipment and procedures for electrolytical phase extraction. We need this equipment for sample preparation for XRD. Thanks in advance.
A colleague has to serial section samples embedded in Araldite. He is trimming the block using a diamond trimming tool and is trimming to a very small block face. The problem is that he is not getting ribbons formed when he sections. Does anyone have a trick to get the sections to stick together and form ribbons? Thanks in advance.
Bob Temkin
Advanced Bioimaging Centre Mount Sinai Hospital Pathology and Laboratory Medicine
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Bill, We too have used the method you've described, but I am not familiar with "glow discharge the grids". What is tis and how does it work?
Thanks, Lou Ross
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==============================Original Headers============================== 6, 19 -- From RossLM-at-missouri.edu Thu Jun 1 13:44:31 2006 6, 19 -- Received: from um-nsmtpout1.um.umsystem.edu (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k51IiUUf020636 6, 19 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 13:44:31 -0500 6, 19 -- Received: from um-exproto9.um.umsystem.edu ([207.160.151.49]) by um-nsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 6, 19 -- Thu, 1 Jun 2006 13:44:30 -0500 6, 19 -- Received: from [128.206.78.127] ([128.206.78.39]) by um-exproto9.um.umsystem.edu over TLS secured channel with Microsoft SMTPSVC(6.0.3790.1830); 6, 19 -- Thu, 1 Jun 2006 13:44:30 -0500 6, 19 -- Mime-Version: 1.0 6, 19 -- X-Sender: RossLM-at-pop.missouri.edu 6, 19 -- Message-Id: {p05200f2fc0a4e55028d2-at-[128.206.78.127]} 6, 19 -- In-Reply-To: {200606011711.k51HBix4023116-at-ns.microscopy.com} 6, 19 -- References: {200606011711.k51HBix4023116-at-ns.microscopy.com} 6, 19 -- Date: Thu, 1 Jun 2006 13:44:29 -0500 6, 19 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 6, 19 -- From: Lou Ross {RossLM-at-missouri.edu} 6, 19 -- Subject: [Microscopy] Re: Sample prep for disbersing powder samples 6, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 6, 19 -- X-OriginalArrivalTime: 01 Jun 2006 18:44:30.0740 (UTC) FILETIME=[6AFB6540:01C685AB] ==============================End of - Headers==============================
I would like to be short....."glow discharging" is a kind of creating a kind of "air plasma" in order to get formvar-coated grids stickier (= more hydrophilic, discharging of surface charge).
This process necessarily /usually is done in a sputter coater or "plasma etcher" or the like (must have a recipient and vacuum pump(s) rotary, and - if (perhaps) you need higher vac, diffusion oil or tubomolecular pump...) as well as an inbuilt ionizing source.
Often such a technique is used for getting formvar-coated grids (used for negative staining procedure on virus particles) again hydrophilic as well as stickier for the adsorption of virus particles to the formvar film. I enclose here - FY convenience - a thread on that issue published in Microscopy Today (and therefore was a theme via the MSA Listserver in 2005.........
NetNotes aus MICROSCOPY TODAY Vol. 13, No 3, May 2005, p. 64-65
Formvar Grids (de-charging by glow discharging and other methods) ---------------- Q: We have been using Formvar/carbon coated grids for spreading magnetic nano-particles in water and for negative staining. But as you know, an aqueous solution does not spread well on grids because of the charge characteristics on the film surface. The only way I know of for making the film surface more hydrophilic is to do a glow discharge in a sputter coater, but we do not have such a device. I heard treating coated grids with ethanol vapor works, but not for me. Does anyone have any other tricks or suggestions? Hong Yi {hyi-at-emory.edu} 20 Feb 2005 ----------- Answers: Do you have a vacuum evaporator? It's simple to rig it for glow discharge with an inexpensive Tesla coil. A plastic vacuum desiccator, the same Tesla coil, and a rough vacuum source will do the job also. Caroline Schooley {schooley-at-mcn.org} 21 Feb 2005 ------------ I don't think that a splitter coater, old or new, is going to solve your problem. I have not heard of ethanol vapors making a carbon grid more hydrophilic. Carbon coated grids lose their hydrophilic nature as they age and they become more hydrophobic. The process can be "reversed" by (a) exposure to an RF "air" plasma in a small plasma etcher (effect will last 60-90 days) or (b) a thin evaporation of VictawetO onto the grids. Our own studies would suggest that Victawet can keep the grids highly hydrophilic essentially forever (e.g. more than one year). Just remember that it is a phosphate based surfactant so if you are doing elemental analysis work, you might not want to have P showing up in your data. But if you have an ordinary vacuum evaporator and tungsten baskets, and don't have a plasma etcher, you can solve your problem with Victawet. The best bet for having carbon coated grids with the greatest hydrophilic characteristics is to make or purchase your carbon coated grids always "fresh': If the grids are purchased, and their age is uncertain, contact the manufacturer of the carbon coated grids, give them the lot number and then you will know. Charles A. Garber {cgarber-at-2spi.com} 21 Feb 2005 ---------- Another method occasionally used to make C-coated grids hydrophilic is to expose them for some minutes to the direct illumination of a UV lamp. This is done at normal atmospheric pressure, so it needs no vacuum technology. James Chalcroft {jchalcro-at-neuro.mpg.de} 21 Feb 2005 ---------- I personally don't like glow discharge at all: it's very difficult to reproduce. It depends on the equipment and there is no way to control the "amount" of discharge. I use 0.5-1 % poly-lysine from any of the EM suppliers (don't try to make the solution yourself since there is some trick required). Place the EM grid on a 10 ?l poly-lysine drop for 5-10 min, wash on a few drops of deionized water, air dry - good for at least a month. Alcian Blue works in the similar way with a similar result. Of course, these methods will only work for positively charged molecules. Sergey Ryazantsev {sryazant-at-ucla.edu} 22 Feb 2005
hope this answers your question... best regards
Wolfgang MUSS Salzburg, Austria =============================} } { {======================== ---------- Von: RossLM-at-missouri.edu[SMTP:RossLM-at-missouri.edu] Antwort an: RossLM-at-missouri.edu Gesendet: Donnerstag, 01. Juni 2006 20:50 An: W.Muss-at-salk.at Betreff: [Microscopy] Re: Sample prep for disbersing powder samples
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Thanks, Lou Ross
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-- Senior Electron Microscope Specialist Electron Microscopy Core Facility W136 Veterinary Medicine University of Missouri Columbia, MO 65211-5120 (573) 882-4777, fax 884=2227 email: rosslm-at-missouri.edu http://www.emc.missouri.edu
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==============================Original Headers============================== 18, 27 -- From W.Muss-at-salk.at Thu Jun 1 14:48:54 2006 18, 27 -- Received: from Hermes.salk.at (hermes.lks.at [193.170.167.9]) 18, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k51JmqDI031970 18, 27 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 14:48:53 -0500 18, 27 -- Received: from localhost (localhost [127.0.0.1]) 18, 27 -- by hermes.lks.at (Postfix) with ESMTP id 037A65A901F; 18, 27 -- Thu, 1 Jun 2006 21:48:47 +0200 (CEST) 18, 27 -- Received: from hermes.lks.at ([127.0.0.1]) 18, 27 -- by localhost (hermes.lks.local [127.0.0.1]) (amavisd-new, port 10024) 18, 27 -- with ESMTP id 79044-02; Thu, 1 Jun 2006 21:48:46 +0200 (CEST) 18, 27 -- Received: from c1pa003 (unknown [192.168.42.3]) 18, 27 -- by hermes.lks.at (Postfix) with SMTP id 8E8E95A900A; 18, 27 -- Thu, 1 Jun 2006 21:48:46 +0200 (CEST) 18, 27 -- Received: by localhost with Microsoft MAPI; Thu, 1 Jun 2006 21:48:46 +0200 18, 27 -- Message-ID: {01C685C5.28851660.W.Muss-at-salk.at} 18, 27 -- From: MUSS Wolfgang PhD {W.Muss-at-salk.at} 18, 27 -- Reply-To: "W.Muss-at-salk.at" {W.Muss-at-salk.at} 18, 27 -- To: "'microscopy-at-msa.microscopy.com'" {microscopy-at-msa.microscopy.com} 18, 27 -- Subject: [Microscopy] Re: Sample prep for disbersing powder samples 18, 27 -- Date: Thu, 1 Jun 2006 21:48:45 +0200 18, 27 -- Return-Receipt-To: MUSS Wolfgang PhD {W.Muss-at-salk.at} 18, 27 -- Organization: SALK,Salzburger Landeskliniken, Pathologie, EM-Labor 18, 27 -- X-Mailer: Microsoft Internet E-Mail/MAPI - 8.0.0.4211 18, 27 -- MIME-Version: 1.0 18, 27 -- Content-Type: text/plain; charset="us-ascii" 18, 27 -- Content-Transfer-Encoding: 7bit 18, 27 -- X-Virii-Scanned: by Sophos/ClamAV at salk.at ==============================End of - Headers==============================
Bill, We too have used the method you've described, but I am not familiar with "glow discharge the grids". What is tis and how does it work?
Thanks, Lou Ross
--------------------------
Hello Lou,
Essentially, glow discharge is a procedure that you can carry out on a standard vacuum evaporator that is equipped with a high voltage AC input. Unfortunately, not many vac evaporators have this accessory. In operation, specimens are loaded into the chamber and one end of about 8-12 inches of high purity aluminum wire (3-5 mm) is plugged into the HV feedthrough and a large loop is formed to surround the grids. The chamber is pumped down using only the rotary pump until you get approximately 50-100 millitorr vacuum. At this point, you turn up the AC high voltage (using a variAC) and you will generate a plasma similar to what one sees in a sputter coater. The plasma is thought to both clean the grid surface as well as imparting a charge that renders the grid surface (of carbon-coated grids) hydrophilic. The effect lasts about a day--less if in a high humidity environment.
We have been able to approach this technique by placing the grids inside a sputter coater and shielding them from direct line of sight with the target (to prevent coating with metal). The plasma generated will impart a hydrophobic character to the grids. But you may also get metal deposition, so do some trial runs first.
I have been depositing a lot of nanocrystals as you describe by directly depositing them on carbon or silicon substrates. The powders are suspended in acetone, shaken and (after allowing the large particles to settle) a small volume taken up in a micropitettor (about 10-20 microliters). This is then dropped directly onto the grid surface from a distance of 10 or so mm.
I can take a picture of the glow discharge setup if you like, Lou.
JB
} } } } On Jun 1, 2006, at 8:40 AM, lkrupp-at-us.ibm.com wrote: } } } } } I need to make samples disbersed on carbon-coated grids for collecting } } } some } } } EELS standards of various oxides in powder form. I have Maganese, } } } Titiatium, and Ruthenium oxides in various states. Anyone know a good } } } prep } } } that will evenly disburse a thin layer onto a carbon-coated grid? My } } } first } } } thought would be to mix with a solvent and put a drop on a grid, but I } } } figured someone here has probably already done this. } } } } } Dear Leslie, } } My choice would be your first thought--it worked very well for } } river-bottom sediment. I would try various dilutions to find the one } } that gives the desired distribution of particles, and I would } } glow-discharge the grids. If using water leads to aggregation, try an } } organic solvent, and if the particles still have a tendency to } } aggregate, try putting the grid with the drop of solvent in an } } oven--faster evaporation might overcome that tendency. } } Yours, } } Bill Tivol, PhD } } EM Scientist and Manager } } Cryo-Electron Microscopy Facility } } Broad Center, Mail Code 114-96 } } California Institute of Technology } } Pasadena CA 91125 } } (626) 395-8833 } } tivol-at-caltech.edu } } } } } } ==============================Original Headers============================== } } 4, 22 -- From tivol-at-caltech.edu Thu Jun 1 12:10:36 2006 } } 4, 22 -- Received: from outgoing-mail.its.caltech.edu } } (outgoing-mail.its.caltech.edu [131.215.239.19]) } } 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } } ESMTP id k51HAawk020886 } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 } } 12:10:36 -0500 } } 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) } } 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id BEB94366A6 } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 } } 10:10:35 -0700 (PDT) } } 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) } } 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id 3A54510AE0C } } 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 } } 10:10:30 -0700 (PDT) } } 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) } } 4, 22 -- In-Reply-To: {200606011540.k51Fe4m9009135-at-ns.microscopy.com} } } 4, 22 -- References: {200606011540.k51Fe4m9009135-at-ns.microscopy.com} } } 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed } } 4, 22 -- Message-Id: {a6cd43e8d0c28b6d5fc583cdda84c003-at-caltech.edu} } } 4, 22 -- Content-Transfer-Encoding: 7bit } } 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} } } 4, 22 -- Subject: Re: [Microscopy] Sample prep for disbersing powder samples } } 4, 22 -- Date: Thu, 1 Jun 2006 10:21:39 -0700 } } 4, 22 -- To: microscopy-at-msa.microscopy.com } } 4, 22 -- X-Mailer: Apple Mail (2.624) } } 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 } } ==============================End of - Headers============================== } } } -- } Senior Electron Microscope Specialist } Electron Microscopy Core Facility } W136 Veterinary Medicine } University of Missouri } Columbia, MO 65211-5120 } (573) 882-4777, fax 884=2227 } email: rosslm-at-missouri.edu } http://www.emc.missouri.edu } } ==============================Original Headers============================== } 6, 19 -- From RossLM-at-missouri.edu Thu Jun 1 13:44:31 2006 } 6, 19 -- Received: from um-nsmtpout1.um.umsystem.edu } (um-nsmtpout1.um.umsystem.edu [209.106.228.53]) } 6, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k51IiUUf020636 } 6, 19 -- for {microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 13:44:31 -0500 } 6, 19 -- Received: from um-exproto9.um.umsystem.edu } ([207.160.151.49]) by um-nsmtpout1.um.umsystem.edu with Microsoft } SMTPSVC(6.0.3790.1830); } 6, 19 -- Thu, 1 Jun 2006 13:44:30 -0500 } 6, 19 -- Received: from [128.206.78.127] ([128.206.78.39]) by } um-exproto9.um.umsystem.edu over TLS secured channel with Microsoft } SMTPSVC(6.0.3790.1830); } 6, 19 -- Thu, 1 Jun 2006 13:44:30 -0500 } 6, 19 -- Mime-Version: 1.0 } 6, 19 -- X-Sender: RossLM-at-pop.missouri.edu } 6, 19 -- Message-Id: {p05200f2fc0a4e55028d2-at-[128.206.78.127]} } 6, 19 -- In-Reply-To: {200606011711.k51HBix4023116-at-ns.microscopy.com} } 6, 19 -- References: {200606011711.k51HBix4023116-at-ns.microscopy.com} } 6, 19 -- Date: Thu, 1 Jun 2006 13:44:29 -0500 } 6, 19 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} } 6, 19 -- From: Lou Ross {RossLM-at-missouri.edu} } 6, 19 -- Subject: [Microscopy] Re: Sample prep for disbersing powder samples } 6, 19 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" } 6, 19 -- X-OriginalArrivalTime: 01 Jun 2006 18:44:30.0740 (UTC) } FILETIME=[6AFB6540:01C685AB] } ==============================End of - Headers==============================
-- ############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
==============================Original Headers============================== 14, 18 -- From bozzola-at-siu.edu Thu Jun 1 14:51:44 2006 14, 18 -- Received: from abbmta2.siu.edu (abbmta2.siu.edu [131.230.254.206]) 14, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k51Jpidf002075 14, 18 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 14:51:44 -0500 14, 18 -- Received: from [131.230.177.142] (ws177142.microscope.siu.edu [131.230.177.142]) 14, 18 -- by abbmta2.siu.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k51JpeEE022652 14, 18 -- for {Microscopy-at-msa.microscopy.com} ; Thu, 1 Jun 2006 14:51:42 -0500 (CDT) 14, 18 -- Mime-Version: 1.0 14, 18 -- X-Sender: bozzola-at-saluki-mail.siu.edu 14, 18 -- Message-Id: {p06110416c0a4f1670b76-at-[131.230.177.142]} 14, 18 -- In-Reply-To: {200606011846.k51Ik0w7022529-at-ns.microscopy.com} 14, 18 -- References: {200606011846.k51Ik0w7022529-at-ns.microscopy.com} 14, 18 -- Date: Thu, 1 Jun 2006 14:51:44 -0500 14, 18 -- To: Microscopy-at-msa.microscopy.com 14, 18 -- From: "John J. Bozzola" {bozzola-at-siu.edu} 14, 18 -- Subject: [Microscopy] Re: Sample prep for disbersing powder samples 14, 18 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 14, 18 -- X-MASF: 0.00% ==============================End of - Headers==============================
Vendors who recondition LKB 7801 B knife makers please contact me. -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 1, 20 -- From oshel1pe-at-cmich.edu Thu Jun 1 14:54:26 2006 1, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 1, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k51JsP7S011509 1, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 14:54:26 -0500 1, 20 -- Received: from egateb.central.cmich.local ([141.209.15.85]) 1, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k51KTl4l023712 1, 20 -- for {Microscopy-at-microscopy.com} ; Thu, 1 Jun 2006 16:29:47 -0400 1, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egateb.central.cmich.local with Microsoft SMTPSVC(5.0.2195.6713); 1, 20 -- Thu, 1 Jun 2006 15:54:25 -0400 1, 20 -- Mime-Version: 1.0 1, 20 -- Message-Id: {f0623090ac0a4f6398d5a-at-[141.209.160.132]} 1, 20 -- Date: Thu, 1 Jun 2006 15:54:24 -0400 1, 20 -- To: Microscopy-at-microscopy.com 1, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 1, 20 -- Subject: LKB knifemaker service 1, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 1, 20 -- X-OriginalArrivalTime: 01 Jun 2006 19:54:25.0358 (UTC) FILETIME=[2F2B4EE0:01C685B5] 1, 20 -- X-CanItPRO-Stream: default 1, 20 -- X-Spam-Score: -4 () L_EXCH_MF 1, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
} ### apologies for multiple postings } } } } } The Department of Sedimentology and Environmental Geology, } Geoscience Center of the University } of Goettingen, is offering a } } } post-doc position in Sedimentology / Sedimentary Petrology } } } beginning 1st October, 2006. The appointment is for one year with an } option for another six month. } Salary is based on the wage agreement of the German civil service } (75% BAT IIa, approx. 30.000 EUR p.a.). } } The Geoscience Center in Goettingen offers a wide range of } state-of-the-art analytical facilities and an excellent environment } for geological, sedimentological and geochemical research ( } http://www.gzg.uni-goettingen.de ). } We are seeking for a gifted and motivated scientist with a several } years perspective for our research team. } } The successful candidate has finished his/her excellent doctoral } thesis no longer than two years ago. Ongoing and future research } should focus on one of the major research topics of the department, } such as exhumation & erosion, weathering processes, sediment } geochemistry and/or } provenance analysis. During the one-year period offered here a } research proposal should be set up to continue research on soft money. } } Teaching duties comprise sedimentary petrology as well as } sedimentary facies and basin analysis at } various levels, as well as field trips. The University of Goettingen } is attempting to increase the } proportion of women on the scientific staff and strongly encourages } qualified women to apply. } Disabled candidates with equal qualification will be preferred. } } Applicants should send a letter of interest, curriculum vitae, a } brief statement of research interests, list of publications and } contact information for two referees, no later than June 30, 2006, } to Prof. Dr. Hilmar von Eynatten, Department of Sedimentology and } Environmental Geology, Geoscience Center Goettingen, } Goldschmidtstrasse 3, D-37077 Goettingen, Germany } (hilmar.von.eynatten-at-geo.uni-goettingen.de, } http://www.sediment.uni-goettingen.de ). }
==============================Original Headers============================== 4, 20 -- From tamas.mikes-at-geo.uni-goettingen.de Fri Jun 2 09:52:03 2006 4, 20 -- Received: from mailer.gwdg.de (mailer.gwdg.de [134.76.10.26]) 4, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k52Eq205001096 4, 20 -- for {microscopy-at-microscopy.com} ; Fri, 2 Jun 2006 09:52:03 -0500 4, 20 -- Received: from sedi8.gzg.geo.uni-goettingen.de ([134.76.77.171] helo=sedi8.geo.uni-goettingen.de) 4, 20 -- by mailer.gwdg.de with esmtp (Exim 4.60) 4, 20 -- (envelope-from {tamas.mikes-at-geo.uni-goettingen.de} ) 4, 20 -- id 1FmB0I-0006jz-7I 4, 20 -- for microscopy-at-microscopy.com; Fri, 02 Jun 2006 16:52:02 +0200 4, 20 -- Message-Id: {7.0.1.0.0.20060602165219.0227a738-at-geo.uni-goettingen.de} 4, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 4, 20 -- Date: Fri, 02 Jun 2006 16:53:12 +0200 4, 20 -- To: "Microscopy Listserver" {microscopy-at-microscopy.com} 4, 20 -- From: Tamas Mikes {tamas.mikes-at-geo.uni-goettingen.de} 4, 20 -- Subject: post-doc position in sedimentary petrology available 4, 20 -- Mime-Version: 1.0 4, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 4, 20 -- X-Spam-Level: / 4, 20 -- X-Spam-Report: Content analysis: 0.0 points, 6.0 required 4, 20 -- X-Virus-Scanned: (clean) by exiscan+sophie ==============================End of - Headers==============================
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I have a SEM sample of bacteria in milk. Does anyone have any suggestions on how to reduce the milk debris/fat leaving the bacteria intact? I have centrifuged the sample, took from the formed pellet and deposited onto a millipore filter on a pump, buffer washed several times and followed a standard fixation protocol though HMDS. Bacteria of other samples (not in milk) looked great, but the milk sample left way too much debris behind.
Thanks in advance for any suggestions. Lynda Schneider University of Florida
It's that time again! We are in need for volunteers for the M&M meeting in Chicago.
Student volunteers will be paid through a voucher system. Other volunteers will receive benefits according to the hours provided. As soon as you are accepted as a volunteer, your name will be provided to meeting registration. All efforts will be made to place volunteers for sessions and booths that they are interested in.
For more information and a schedule of sessions/jobs, please contact John Shields at jshields-at-cb.uga.edu (706-542-4080), or to Bill Monroe at monroe-at-emcenter.msstate.edu (662-325- 3019). John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602
706-542-4080
==============================Original Headers============================== 4, 22 -- From jpshield-at-uga.edu Mon Jun 5 09:24:38 2006 4, 22 -- Received: from puntd5.cc.uga.edu (puntd5.cc.uga.edu [128.192.1.108]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k55EObbx001319 4, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 5 Jun 2006 09:24:38 -0500 4, 22 -- Received: from punts4.cc.uga.edu (punts4.cc.uga.edu [128.192.1.115]) 4, 22 -- by puntd5.cc.uga.edu (MOS 3.7.4b-GA) 4, 22 -- with ESMTP id CWW21100; 4, 22 -- Mon, 5 Jun 2006 10:24:36 -0400 (EDT) 4, 22 -- Received: (from punts4.cc.uga.edu [128.192.63.198]) 4, 22 -- by punts4.cc.uga.edu (MOS 3.7.4b-GA) 4, 22 -- with HTTPS/1.1 id AVU01645 (AUTH jpshield-at-uga.edu); 4, 22 -- Mon, 5 Jun 2006 10:24:34 -0400 (EDT) 4, 22 -- From: John Shields {jpshield-at-uga.edu} 4, 22 -- Subject: volunteer workers for M&M Chicago 4, 22 -- To: msa listserve {Microscopy-at-MSA.Microscopy.Com} 4, 22 -- Cc: {monroe-at-emcenter.msstate.edu} 4, 22 -- X-Mailer: Mirapoint Webmail Direct 3.7.4b-GA 4, 22 -- MIME-Version: 1.0 4, 22 -- Content-Type: text/plain; charset=us-ascii 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- Message-Id: {20060605102434.AVU01645-at-punts4.cc.uga.edu} 4, 22 -- Date: Mon, 5 Jun 2006 10:24:34 -0400 (EDT) ==============================End of - Headers==============================
We have a grad student user who is interested in thin sectioning carbon nanotubes. The tubes are in clusters and suspended in some buffer. Is it possible to embed the clumps in epoxy resin and section them? Is diamond knife going to survive after sectioning nanotubes? The average diameter of those tubes is approx. 1.4 nm. Any suggestion will be greatly appreciated.
Thanks in advance,
Soumitra
****************************************************************************** Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office) 505-646-3283 (lab) Fax: 505-646-3282 http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu
==============================Original Headers============================== 5, 20 -- From sghoshro-at-nmsu.edu Mon Jun 5 14:13:53 2006 5, 20 -- Received: from cheech.nmsu.edu (cheech.nmsu.edu [128.123.34.14]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k55JDrU9019226 5, 20 -- for {Microscopy-at-microscopy.com} ; Mon, 5 Jun 2006 14:13:53 -0500 5, 20 -- Received: (from nobody-at-localhost) 5, 20 -- by cheech.nmsu.edu (8.11.6/8.11.6) id k55JDrU09264 5, 20 -- for Microscopy-at-microscopy.com; Mon, 5 Jun 2006 13:13:53 -0600 5, 20 -- Received: from 128.123.105.122 ( [128.123.105.122]) 5, 20 -- as user sghoshro-at-imap.nmsu.edu by webmail.nmsu.edu with HTTP; 5, 20 -- Mon, 5 Jun 2006 13:13:53 -0600 5, 20 -- Message-ID: {1149534833.44848271153f4-at-webmail.nmsu.edu} 5, 20 -- Date: Mon, 5 Jun 2006 13:13:53 -0600 5, 20 -- From: Soumitra Ghoshroy {sghoshro-at-nmsu.edu} 5, 20 -- To: Microscopy-at-microscopy.com 5, 20 -- Subject: sectioning carbon nanotubes 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; charset=ISO-8859-1 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- User-Agent: Internet Messaging Program (IMP) 3.0 5, 20 -- X-Originating-IP: 128.123.105.122 ==============================End of - Headers==============================
We are planning to examine some cellulose made microspheres containing albumin with transmission EM. Is there anybody experienced with this material before? Are routine EM tissue processing methods suitable with this material? Thanks for any kind of helps...
Dr. Necat Yilmaz Mersin Universitesi Tip Fakultesi Histoloji ve Embriyoloji Anabilim Dali
==============================Original Headers============================== 4, 31 -- From nyilmaz-at-mersin.edu.tr Tue Jun 6 07:32:34 2006 4, 31 -- Received: from mail.mersin.edu.tr (mail.mersin.edu.tr [193.255.128.3]) 4, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k56CWVBm020888 4, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 6 Jun 2006 07:32:34 -0500 4, 31 -- Received: from localhost (localhost.mersin.edu.tr [127.0.0.1]) 4, 31 -- by mail.mersin.edu.tr (Postfix) with ESMTP id 2AFA245146 4, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 6 Jun 2006 15:32:34 +0300 (EEST) 4, 31 -- Received: from mail.mersin.edu.tr ([127.0.0.1]) 4, 31 -- by localhost (mail.mersin.edu.tr [127.0.0.1]) (amavisd-new, port 10024) 4, 31 -- with ESMTP id 86250-05 for {Microscopy-at-microscopy.com} ; 4, 31 -- Tue, 6 Jun 2006 15:32:21 +0300 (EEST) 4, 31 -- Received: from NEJAT1 (unknown [193.255.128.130]) 4, 31 -- by mail.mersin.edu.tr (Postfix) with SMTP id 176D84513A 4, 31 -- for {Microscopy-at-microscopy.com} ; Tue, 6 Jun 2006 15:32:17 +0300 (EEST) 4, 31 -- Message-ID: {002e01c68965$3c9f8fb0$5201a8c0-at-NEJAT1} 4, 31 -- From: "Nejat" {nyilmaz-at-mersin.edu.tr} 4, 31 -- To: "EM-Mail Group" {Microscopy-at-microscopy.com} 4, 31 -- Subject: Help for cellulose microsphere processing 4, 31 -- Date: Tue, 6 Jun 2006 15:32:06 +0300 4, 31 -- MIME-Version: 1.0 4, 31 -- Content-Type: text/plain; 4, 31 -- format=flowed; 4, 31 -- charset="windows-1254"; 4, 31 -- reply-type=original 4, 31 -- Content-Transfer-Encoding: 7bit 4, 31 -- X-Priority: 3 4, 31 -- X-MSMail-Priority: Normal 4, 31 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 4, 31 -- Disposition-Notification-To: "Nejat" {nyilmaz-at-mersin.edu.tr} 4, 31 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 4, 31 -- X-Virus-Scanned: by amavisd-new at mersin.edu.tr ==============================End of - Headers==============================
We solved our contrasting problems by just following simple rules found in this list. If it solved our problems, I thought it could be useful for others.
- We prepared "carbonated lead citrate" (the time required to "burn" it was much shorter as proposed though) - We bought decarbonated NaOH to a big american supplier
These are very simple rules which make all the difference between "I see nothing" and "oh my god I never had such a nice staining". And that's true: I've never ever had such a beautiful staining!
Thank you again for your support.
Stéphane
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==============================Original Headers============================== 7, 18 -- From nizets2-at-yahoo.com Tue Jun 6 09:10:06 2006 7, 18 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.87.65]) 7, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k56EA6M4002486 7, 18 -- for {microscopy-at-microscopy.com} ; Tue, 6 Jun 2006 09:10:06 -0500 7, 18 -- Received: (qmail 2845 invoked by uid 60001); 6 Jun 2006 14:10:06 -0000 7, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 7, 18 -- s=s1024; d=yahoo.com; 7, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 7, 18 -- b=nySdMzGji0kCEsaiYe3tpvougj/SVOO9ewwIWzMh2PBKN2eoYk1WF8iHp+JGNMcn+ly4RP7tBbHr5wlMS6iU4P42BsOTjHxoB1QGGLAU7XQL7doeX+fDLkwpvBBIcPd7VR0Rka5PTOGHmO3EMGBktWvXGdeK5WnyfMEqD6FpV5o= ; 7, 18 -- Message-ID: {20060606141006.2837.qmail-at-web37412.mail.mud.yahoo.com} 7, 18 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Tue, 06 Jun 2006 07:10:06 PDT 7, 18 -- Date: Tue, 6 Jun 2006 07:10:06 -0700 (PDT) 7, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 7, 18 -- Subject: Lead citrate staining: Thank you again and again.... 7, 18 -- To: microscopy-at-microscopy.com 7, 18 -- MIME-Version: 1.0 7, 18 -- Content-Type: text/plain; charset=iso-8859-1 7, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Here is another weird idea from me :-D I would like to localize a fluorescent molecule in the transversal plane of a cell monolayer(not from above). We have no material for the preparation of histological sections, but we have all necessary for TEM. I wonder if I could not embed the monolayer in Epon, then cut transversal semi-thin sections and observe in epifluroscence. Would Epon hinder fluorescence? Could it be done in another resin than Epon?
Regards,
Stéphane
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==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Wed Jun 7 03:03:17 2006 6, 18 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.87.63]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5783Hql015897 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 03:03:17 -0500 6, 18 -- Received: (qmail 21951 invoked by uid 60001); 7 Jun 2006 08:03:17 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.com; 6, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; 6, 18 -- b=mYPALc5dKwVqDFpaUMA79vMBlV2nctfE6rU9oK5u9cklXlbjaQfseuZYkqBTWh9dMv2Mk7fPbVFk87hXzbRCNMvxud5I4FFYAxJ+K7wDw3vTWrY/0foDb8GCrPhQjIEvPHM78A2MPWGG3n90jptl6XsGrL3IO0Qs+45yxbbeZT0= ; 6, 18 -- Message-ID: {20060607080317.21949.qmail-at-web37410.mail.mud.yahoo.com} 6, 18 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 01:03:17 PDT 6, 18 -- Date: Wed, 7 Jun 2006 01:03:17 -0700 (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: weird idea: fluorescence in Epon 6, 18 -- To: microscopy-at-microscopy.com 6, 18 -- MIME-Version: 1.0 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 6, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I would appreciate any comments, references, or first-hand experiences from those using a combined FIB/SEM+Cryo instrument with & without a field emission gun - where biological or polymer samples can be sliced by the ion beam then imaged. I would be interested to know things such as:
Why you chose the instrument you use? How easy is it in practice to do this sort of ion beam slicing & is it restricted to particular types of biological or polymer samples? Does the FIB mode impact on the column cleanliness & resolution over time particularly if used to thin samples for TEM? Is the instrument expensive to run & does it require an experienced operator? Anything else you feel would help those looking at this type of instrument for a multi-user facility.
Many thanks Ursula -------------------
Ursula J. Potter Centre for Electron Optical Studies (CEOS) Building 3 West 2.15 The University of Bath Claverton Down Bath BA2 7AY UK Tel: 01225 385651 Email: U.J.Potter-at-bath.ac.uk
==============================Original Headers============================== 6, 23 -- From U.J.Potter-at-bath.ac.uk Wed Jun 7 05:11:33 2006 6, 23 -- Received: from binda.bath.ac.uk (binda.bath.ac.uk [138.38.32.22]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k57ABWSl029055 6, 23 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 05:11:33 -0500 6, 23 -- Received: from mary.bath.ac.uk ([138.38.32.14] ident=mmdf) 6, 23 -- by binda.bath.ac.uk with smtp id 1Fnv0e-0001sg-A3 6, 23 -- for microscopy-at-microscopy.com 6, 23 -- (return-path {U.J.Potter-at-bath.ac.uk} ); Wed, 07 Jun 2006 11:11:32 +0100 6, 23 -- Received: from eapc-03.campus.bath.ac.uk 6, 23 -- ( eapc-03.campus.bath.ac.uk [138.38.136.63] ) by bath.ac.uk 6, 23 -- id ab27759 for {microscopy-at-microscopy.com} ; 7 Jun 2006 11:11 +0100 6, 23 -- Date: Wed, 07 Jun 2006 11:11:24 +0100 6, 23 -- From: Ursula Potter {U.J.Potter-at-bath.ac.uk} 6, 23 -- To: microscopy-at-microscopy.com 6, 23 -- Subject: FIB/SEM+Cryo 6, 23 -- Message-ID: {7688765.1149678684-at-eapc-03.campus.bath.ac.uk} 6, 23 -- Originator-Info: login-id=mssujp; server=imaphost.bath.ac.uk 6, 23 -- X-Mailer: Mulberry/3.1.0 (Win32) 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; charset=us-ascii; FORMAT=flowed 6, 23 -- Content-Transfer-Encoding: 7bit 6, 23 -- Content-Disposition: inline 6, 23 -- X-Scanner: 207b733ce87962cd23b7af89110027a7b5b8d829 ==============================End of - Headers==============================
Are there anyone out there using this model? I would need specially the low level protocol on commanding it's RS232C port for lab automation purposes on our lab set-up at the university. Your help is greatly appreciated.
Regards,
Berns B.
-- -------------------------------------------- Bernardino Jerez Buenaobra University Research Associate National Institute of Physics University of the Philippines Diliman Campus 1101 Quezon City Philippines VOIP : +6329205301-99 local 3703 Fax/Data: +6329280296 URL: http://www.nip.upd.edu.ph/ipl email: bbuenaobra-at-nip.upd.edu.ph --------------------------------------------------------------------------
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Stephane, Not so weird, but might not work. The two things to worry about are autofluorescence and loss of your fluorochrome. If you have some of your cells (unlabeled) in epon you can check some sections under a fluorescence microscope and see about the autofuor. I think the loss of fluorescence is a more serious problem, either because it is extracted during dehydration/infiltration or quenched by the embedment. You may have to embed a labeled sample and find out the hard way!
Tobias } } Dear listers, } } Here is another weird idea from me :-D } I would like to localize a fluorescent molecule in the } transversal plane of a cell monolayer(not from above). } We have no material for the preparation of } histological sections, but we have all necessary for } TEM. I wonder if I could not embed the monolayer in } Epon, then cut transversal semi-thin sections and } observe in epifluroscence. } Would Epon hinder fluorescence? } Could it be done in another resin than Epon? } } Regards, } } StÈphane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Wed Jun 7 03:03:17 2006 } 6, 18 -- Received: from } web37410.mail.mud.yahoo.com } (web37410.mail.mud.yahoo.com [209.191.87.63]) } 6, 18 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with SMTP id } k5783Hql015897 } 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 03:03:17 -0500 } 6, 18 -- Received: (qmail 21951 invoked by uid } 60001); 7 Jun 2006 08:03:17 -0000 } 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 6, 18 -- s=s1024; d=yahoo.com; } 6, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 6, 18 -- } b=mYPALc5dKwVqDFpaUMA79vMBlV2nctfE6rU9oK5u9cklXlbjaQfseuZYkqBTWh9dMv2Mk7fPbVFk87hXzbRCNMvxud5I4FFYAxJ+K7wDw3vTWrY/0foDb8GCrPhQjIEvPHM78A2MPWGG3n90jptl6XsGrL3IO0Qs+45yxbbeZT0= } ; } 6, 18 -- Message-ID: {20060607080317.21949.qmail-at-web37410.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by } web37410.mail.mud.yahoo.com via HTTP; Wed, 07 } Jun 2006 01:03:17 PDT } 6, 18 -- Date: Wed, 7 Jun 2006 01:03:17 -0700 (PDT) } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 6, 18 -- Subject: weird idea: fluorescence in Epon } 6, 18 -- To: microscopy-at-microscopy.com } 6, 18 -- MIME-Version: 1.0 } 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 6, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers==============================
Regarding the idea of localizing fluorescence label in epon, I would suggest that you not use Epon...there are autoflourescence problems (at least in our hands, using Spurrs resin). However, I think you may be happy using LR White, polymerized at 60C. We've done some similar work here and have been very impressed with the results.
Good luck,
Doug
Douglas R. Keene Assistant Investigator Micro-Imaging Center Shriners Hospital for Children 3102 S.W. Sam Jackson Park Road Portland, Oregon 97239 503-221-3434 drk-at-shcc.org -----Original Message----- X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] Sent: Wednesday, June 07, 2006 1:10 AM To: drk-at-SHCC.org
Dear listers,
Here is another weird idea from me :-D I would like to localize a fluorescent molecule in the transversal plane of a cell monolayer(not from above). We have no material for the preparation of histological sections, but we have all necessary for TEM. I wonder if I could not embed the monolayer in Epon, then cut transversal semi-thin sections and observe in epifluroscence. Would Epon hinder fluorescence? Could it be done in another resin than Epon?
Regards,
Stéphane
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 6, 18 -- From nizets2-at-yahoo.com Wed Jun 7 03:03:17 2006 6, 18 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.87.63]) 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5783Hql015897 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 03:03:17 -0500 6, 18 -- Received: (qmail 21951 invoked by uid 60001); 7 Jun 2006 08:03:17 -0000 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 6, 18 -- s=s1024; d=yahoo.com; 6, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content -Transfer-Encoding; 6, 18 -- b=mYPALc5dKwVqDFpaUMA79vMBlV2nctfE6rU9oK5u9cklXlbjaQfseuZYkqBTWh9dMv2Mk7fPbV Fk87hXzbRCNMvxud5I4FFYAxJ+K7wDw3vTWrY/0foDb8GCrPhQjIEvPHM78A2MPWGG3n90jptl6X sGrL3IO0Qs+45yxbbeZT0= ; 6, 18 -- Message-ID: {20060607080317.21949.qmail-at-web37410.mail.mud.yahoo.com} 6, 18 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 01:03:17 PDT 6, 18 -- Date: Wed, 7 Jun 2006 01:03:17 -0700 (PDT) 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} 6, 18 -- Subject: weird idea: fluorescence in Epon 6, 18 -- To: microscopy-at-microscopy.com 6, 18 -- MIME-Version: 1.0 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 6, 18 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 16, 22 -- From drk-at-SHCC.org Wed Jun 7 12:46:36 2006 16, 22 -- Received: from shcc.org (mail.shcc.org [64.213.211.200]) 16, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k57HkZdU005537 16, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 7 Jun 2006 12:46:35 -0500 16, 22 -- Received: from DRK2 ("port 1395"-at-[64.213.211.3]) by SHCC.org (PMDF V6.1 #37411) 16, 22 -- with ESMTPA id {01M3CA5GN1460004DI-at-SHCC.org} for Microscopy-at-Microscopy.Com; 16, 22 -- Wed, 07 Jun 2006 10:50:12 -0800 (PST) 16, 22 -- Date: Wed, 07 Jun 2006 10:43:17 -0700 16, 22 -- From: Doug Keene {drk-at-SHCC.org} 16, 22 -- Subject: RE: [Microscopy] weird idea: fluorescence in Epon 16, 22 -- In-reply-to: {200606070809.k5789kOv024103-at-ns.microscopy.com} 16, 22 -- To: Microscopy-at-Microscopy.Com 16, 22 -- Reply-to: drk-at-SHCC.org 16, 22 -- Message-id: {01M3CA5GPO8O0004DI-at-SHCC.org} 16, 22 -- Organization: Shriners Hospitals for Children 16, 22 -- MIME-version: 1.0 16, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 16, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 16, 22 -- Content-type: text/plain; charset=iso-8859-1 16, 22 -- Thread-Index: AcaKCQiVuqpcDFhbQBCvgNSUNvkSUwAUE+Gg 16, 22 -- Content-Transfer-Encoding: 8bit 16, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k57HkZdU005537 ==============================End of - Headers==============================
Don't ever hesitate. some of my best things have happened because of an offbeat idea or insane shot in the dark question during discussions. Of course, I don't want to admit that to my wife. She hates to be referred to as an offbeat idea, just as the best thing that ever happened to me O:-) .
Frankly, you're keeping the rest of us fresh and on our toes.
Paul
==============================Original Headers============================== 4, 21 -- From paul_hazelton-at-umanitoba.ca Wed Jun 7 13:31:24 2006 4, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k57IVNQJ016492 4, 21 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 13:31:24 -0500 4, 21 -- Received: from [130.179.152.79] (cvx-016.cc.umanitoba.ca [130.179.152.79]) 4, 21 -- (authenticated bits=0) 4, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k57IVLmc025694; 4, 21 -- Wed, 7 Jun 2006 13:31:22 -0500 (CDT) 4, 21 -- Message-ID: {44871B76.7060704-at-umanitoba.ca} 4, 21 -- Date: Wed, 07 Jun 2006 13:31:18 -0500 4, 21 -- From: Paul Hazelton {paul_hazelton-at-umanitoba.ca} 4, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) Gecko/20040804 Netscape/7.2 (ax) 4, 21 -- X-Accept-Language: en-us, en 4, 21 -- MIME-Version: 1.0 4, 21 -- To: nizets2-at-yahoo.com 4, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 4, 21 -- Subject: Re: [Microscopy] weird idea: fluorescence in Epon 4, 21 -- References: {200606070806.k5786Xow019680-at-ns.microscopy.com} 4, 21 -- In-Reply-To: {200606070806.k5786Xow019680-at-ns.microscopy.com} 4, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 21 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Dear without an I, Success will depend upon the fluorophore. FITC and TRITC will lose their fluorescence. Alexa fluor dyes and cyanine dyes will work well. Actually, I think with reduced photobleaching. Autofluorescence will depend upon imaging modality. Spurr's looks great under the confocal, but the autolfuorescence creates an impossible haze with epi-fluorescence on thick samples. Deconvolution will clean up the haze. 3 micron sections looked very good. the new formulation of spurr's is too brittle so I'm going to try Eponate 812 or something similar for my next round of embedded fluorescence. I tried Histo-Resin and found the autofluorescence was noticeable under confocal, but didn't try LR-White.
Regards, Glen On Jun 7, 2006, at 10:49 AM, drk-at-SHCC.org wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Regarding the idea of localizing fluorescence label in epon, I } would suggest } that you not use Epon...there are autoflourescence problems (at } least in our } hands, using Spurrs resin). However, I think you may be happy } using LR } White, polymerized at 60C. We've done some similar work here and } have been } very impressed with the results. } } Good luck, } } Doug } } Douglas R. Keene } Assistant Investigator } Micro-Imaging Center } Shriners Hospital for Children } 3102 S.W. Sam Jackson Park Road } Portland, Oregon 97239 } 503-221-3434 } drk-at-shcc.org } -----Original Message----- } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } Sent: Wednesday, June 07, 2006 1:10 AM } To: drk-at-SHCC.org } Subject: [Microscopy] weird idea: fluorescence in Epon } } } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } Dear listers, } } Here is another weird idea from me :-D } I would like to localize a fluorescent molecule in the } transversal plane of a cell monolayer(not from above). } We have no material for the preparation of } histological sections, but we have all necessary for } TEM. I wonder if I could not embed the monolayer in } Epon, then cut transversal semi-thin sections and } observe in epifluroscence. } Would Epon hinder fluorescence? } Could it be done in another resin than Epon? } } Regards, } } Stéphane } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original } Headers============================== } 6, 18 -- From nizets2-at-yahoo.com Wed Jun 7 03:03:17 2006 } 6, 18 -- Received: from web37410.mail.mud.yahoo.com } (web37410.mail.mud.yahoo.com [209.191.87.63]) } 6, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id } k5783Hql015897 } 6, 18 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 03:03:17 } -0500 } 6, 18 -- Received: (qmail 21951 invoked by uid 60001); 7 Jun 2006 } 08:03:17 } -0000 } 6, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 6, 18 -- s=s1024; d=yahoo.com; } 6, 18 -- } h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content- } Type:Content } -Transfer-Encoding; } 6, 18 -- } b=mYPALc5dKwVqDFpaUMA79vMBlV2nctfE6rU9oK5u9cklXlbjaQfseuZYkqBTWh9dMv2M } k7fPbV } Fk87hXzbRCNMvxud5I4FFYAxJ+K7wDw3vTWrY/ } 0foDb8GCrPhQjIEvPHM78A2MPWGG3n90jptl6X } sGrL3IO0Qs+45yxbbeZT0= ; } 6, 18 -- Message-ID: } {20060607080317.21949.qmail-at-web37410.mail.mud.yahoo.com} } 6, 18 -- Received: from [80.122.101.102] by } web37410.mail.mud.yahoo.com via } HTTP; Wed, 07 Jun 2006 01:03:17 PDT } 6, 18 -- Date: Wed, 7 Jun 2006 01:03:17 -0700 (PDT) } 6, 18 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 6, 18 -- Subject: weird idea: fluorescence in Epon } 6, 18 -- To: microscopy-at-microscopy.com } 6, 18 -- MIME-Version: 1.0 } 6, 18 -- Content-Type: text/plain; charset=iso-8859-1 } 6, 18 -- Content-Transfer-Encoding: 8bit } ==============================End of - } Headers============================== } } } } ==============================Original } Headers============================== } 16, 22 -- From drk-at-SHCC.org Wed Jun 7 12:46:36 2006 } 16, 22 -- Received: from shcc.org (mail.shcc.org [64.213.211.200]) } 16, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k57HkZdU005537 } 16, 22 -- for {Microscopy-at-Microscopy.Com} ; Wed, 7 Jun 2006 } 12:46:35 -0500 } 16, 22 -- Received: from DRK2 ("port 1395"-at-[64.213.211.3]) by } SHCC.org (PMDF V6.1 #37411) } 16, 22 -- with ESMTPA id {01M3CA5GN1460004DI-at-SHCC.org} for } Microscopy-at-Microscopy.Com; } 16, 22 -- Wed, 07 Jun 2006 10:50:12 -0800 (PST) } 16, 22 -- Date: Wed, 07 Jun 2006 10:43:17 -0700 } 16, 22 -- From: Doug Keene {drk-at-SHCC.org} } 16, 22 -- Subject: RE: [Microscopy] weird idea: fluorescence in Epon } 16, 22 -- In-reply-to: {200606070809.k5789kOv024103-at-ns.microscopy.com} } 16, 22 -- To: Microscopy-at-Microscopy.Com } 16, 22 -- Reply-to: drk-at-SHCC.org } 16, 22 -- Message-id: {01M3CA5GPO8O0004DI-at-SHCC.org} } 16, 22 -- Organization: Shriners Hospitals for Children } 16, 22 -- MIME-version: 1.0 } 16, 22 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } 16, 22 -- X-Mailer: Microsoft Office Outlook, Build 11.0.5510 } 16, 22 -- Content-type: text/plain; charset=iso-8859-1 } 16, 22 -- Thread-Index: AcaKCQiVuqpcDFhbQBCvgNSUNvkSUwAUE+Gg } 16, 22 -- Content-Transfer-Encoding: 8bit } 16, 22 -- X-MIME-Autoconverted: from quoted-printable to 8bit by } ns.microscopy.com id k57HkZdU005537 } ==============================End of - } Headers==============================
==============================Original Headers============================== 6, 25 -- From glenmac-at-u.washington.edu Wed Jun 7 15:36:08 2006 6, 25 -- Received: from mxout7.cac.washington.edu (mxout7.cac.washington.edu [140.142.32.178]) 6, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k57Ka8xv029950 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 15:36:08 -0500 6, 25 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.32.139]) 6, 25 -- by mxout7.cac.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k57Ka6Qu010939 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 13:36:07 -0700 6, 25 -- X-Auth-Received: from [128.95.178.71] ([128.95.178.71]) 6, 25 -- (authenticated authid=glenmac) 6, 25 -- by smtp.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k57Ka6FG007766 6, 25 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 6, 25 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 13:36:06 -0700 6, 25 -- Mime-Version: 1.0 (Apple Message framework v750) 6, 25 -- In-Reply-To: {200606071749.k57HnXYf008396-at-ns.microscopy.com} 6, 25 -- References: {200606071749.k57HnXYf008396-at-ns.microscopy.com} 6, 25 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 6, 25 -- Message-Id: {B6197299-29B8-44FB-9547-C7E54AE7E765-at-u.washington.edu} 6, 25 -- From: Glen MacDonald {glenmac-at-u.washington.edu} 6, 25 -- Subject: RE: weird idea 6, 25 -- Date: Wed, 7 Jun 2006 13:36:03 -0700 6, 25 -- To: microscopy-at-microscopy.com 6, 25 -- X-Mailer: Apple Mail (2.750) 6, 25 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='IP_HTTP_ADDR 0, __C230066_P5 0, __CP_NAME_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __FRAUD_419_FUNWORDS 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0, __STOCK_PHRASE_24 0' 6, 25 -- Content-Transfer-Encoding: 8bit 6, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k57Ka8xv029950 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both wim.vandenbroeck-at-UGent.be as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: wim.vandenbroeck-at-UGent.be Name: Wim Van den Broeck
Organization: Morphology, Ghent University
Title-Subject: [Filtered] TEM - virus particles in cell culture
Question: Dear Friends,
I need to visualize viral particles grown in cell culture, but I do not have any experience with that (only with tissue samples). Could you give me any advice (collection of cells, fixation, embedding, Ö.).
Thanks in advance,
Wim Van den Broeck.
Wim Van den Broeck, DVM, MSc, PhD Professor in Cytology and Histology Department of Morphology Faculty of Veterinary Medicine, Ghent University Salisburylaan 133, B-9820 Merelbeke, BELGIUM tel.: +32 (0)9 264 77 16 fax: +32 (0)9 264 77 90 Email: wim.vandenbroeck-at-UGent.be
While on the Medical School Faculty of USC, Los Angeles I did extensive studies of hepatitis B virus in liver explants. The technique is straightforward.
Remove cells using a soft spatula or a pipette depending on whether or not the cells are attached to the surface of the culture container.
Place the cells in a centrifuge tube containg fixative. Carry out the dehydration and embedding fluid impregnation process in the centrifuge tube spinning between each stage. Finally transfer the embedding mixture with the cells into a Beem-type embedding capsule and spin the cells down into the tip.
If you wish I can send you the reference for one of my publications that outlines the techniques employed.
Best wishes,
Ted Dunn The EMscope Company Ltd. Thailand
--- wim.vandenbroeck-at-UGent.be wrote:
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==============================Original Headers============================== 12, 20 -- From drteddunne-at-yahoo.com Wed Jun 7 23:57:20 2006 12, 20 -- Received: from web33409.mail.mud.yahoo.com (web33409.mail.mud.yahoo.com [68.142.206.141]) 12, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k584vKhx028460 12, 20 -- for {microscopy-at-microscopy.com} ; Wed, 7 Jun 2006 23:57:20 -0500 12, 20 -- Received: (qmail 54631 invoked by uid 60001); 8 Jun 2006 04:57:19 -0000 12, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 12, 20 -- s=s1024; d=yahoo.com; 12, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 12, 20 -- b=ii5iIj/zzCWEX2M/ngtTuKWaGi8Z+F/IefIeH2PH3lxLPeW1epwoduWCMBPKHXXdMNBhf+QT33GPjKZYFC7TkD2ZIvkyySuyJZb+Ee3LkP4ReWtMu81pX1SxE14EKdXGKwtDlOvan5i90MACA9VNLQLULr+BoQvcXeaygnrVzIk= ; 12, 20 -- Message-ID: {20060608045719.54629.qmail-at-web33409.mail.mud.yahoo.com} 12, 20 -- Received: from [202.47.247.136] by web33409.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 21:57:19 PDT 12, 20 -- Date: Wed, 7 Jun 2006 21:57:19 -0700 (PDT) 12, 20 -- From: ted dunn {drteddunne-at-yahoo.com} 12, 20 -- Subject: Re: [Microscopy] viaWWW: TEM - virus particles in cell culture 12, 20 -- To: wim.vandenbroeck-at-UGent.be 12, 20 -- Cc: microscopy-at-microscopy.com 12, 20 -- In-Reply-To: {200606080040.k580eZFL018920-at-ns.microscopy.com} 12, 20 -- MIME-Version: 1.0 12, 20 -- Content-Type: text/plain; charset=iso-8859-1 12, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
It depends if you need to visualize the viral particles inside the cells or not. If so, I would consider usual protocol for classical cell morphology.
I already observed viral particles in cells I observed for other purposes. If you need the viral particles alone, you have 2 solutions depending on the concentration of the virus in medium.
1) If it is concentrated, you just collect the supernatant, centrifuge 5 min at 5000 RPM to pellet the cells and cell debris, and collect the supernatant again.
2) If it is diluted, you have to find an ultrafast centrifuge and perform an additional centrifugation to pellet your virus particles and concentrate them.
Then you can just do a negative staining (PTA worked well with rhinoviruses for me). You will find tons of protocols on the net.
Good luck.
Stephane
--- wim.vandenbroeck-at-UGent.be wrote:
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==============================Original Headers============================== 13, 20 -- From nizets2-at-yahoo.com Thu Jun 8 01:33:06 2006 13, 20 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.87.63]) 13, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k586X6Rv008226 13, 20 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 01:33:06 -0500 13, 20 -- Received: (qmail 55709 invoked by uid 60001); 8 Jun 2006 06:33:05 -0000 13, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 13, 20 -- s=s1024; d=yahoo.com; 13, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 13, 20 -- b=Fsb/2635sW1xpqYSJnpNsxRuXjWSQC4ZSMEQFn3Yquyv/aPwzxX0FRbUXkLKZbkhxW68SgzbmFvBvPPlenBsbYoFrfcTMqb1AEv3c+TGRtOO04pJOKy2iFrdunIgjwMtvkzWuNbp9OESpfyRU5/X8aoheanFV5OzQL4k8yq9fWU= ; 13, 20 -- Message-ID: {20060608063305.55707.qmail-at-web37410.mail.mud.yahoo.com} 13, 20 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 23:33:05 PDT 13, 20 -- Date: Wed, 7 Jun 2006 23:33:05 -0700 (PDT) 13, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 13, 20 -- Subject: Re: [Microscopy] viaWWW: TEM - virus particles in cell culture 13, 20 -- To: wim.vandenbroeck-at-UGent.be 13, 20 -- Cc: microscopy-at-microscopy.com 13, 20 -- In-Reply-To: {200606080042.k580gPDu021981-at-ns.microscopy.com} 13, 20 -- MIME-Version: 1.0 13, 20 -- Content-Type: text/plain; charset=iso-8859-1 13, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
For those interested in the technique, it is simply available in the internet site of EMS. Look in Technical-} tips and articles--} "A stable lead staining solution".
For the very practical side of the preparation of carbonated lead citrate, here is what I exactly did: I heated 2g at 250°C for 3h in a ceramic beaker (called "melting pot" on EMS site) with a glass cover. I have the chance to work with chemists so the equipment is not a problem: I used a temperature-controlled special oven connected to a mobile hood.
I controlled the color of the powder. It turns grey, then yellowish grey, a little bit like humid sand. I stopped then. If one is unsure, one can still heat several beakers each containing 1g of lead citrate and heat them for different times (2h, 3h, 4h).
I think it is pretty straithforward because it worked the first time I tried. I stored the powder in a sealed glass flask (don't know if it is the best though).
Now IMHO, seeing the easiness of the technique and the cheap price of carbonate-free NaOH, there is really no reason NOT to do it :-D
Stephane
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==============================Original Headers============================== 8, 19 -- From nizets2-at-yahoo.com Thu Jun 8 01:55:43 2006 8, 19 -- Received: from web37412.mail.mud.yahoo.com (web37412.mail.mud.yahoo.com [209.191.87.65]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k586thms018635 8, 19 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 01:55:43 -0500 8, 19 -- Received: (qmail 55159 invoked by uid 60001); 8 Jun 2006 06:55:43 -0000 8, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 19 -- s=s1024; d=yahoo.com; 8, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 19 -- b=HSwCug+LY+IF9G7UfVpv3nug1PPUOuKu932Mci/MLXwwynTrFZBvG/o4Fi3NLhxTH7BOceQ0qBQfwoAbazPN3jdO08n6Ux3BTmfGzWngftdTk9q0wgb4vzW4h+aWlkz5SlhxyofQMtJimVunPbuxbr/q6gUn8x9QmTJmDfyIHW8= ; 8, 19 -- Message-ID: {20060608065543.55157.qmail-at-web37412.mail.mud.yahoo.com} 8, 19 -- Received: from [80.122.101.102] by web37412.mail.mud.yahoo.com via HTTP; Wed, 07 Jun 2006 23:55:43 PDT 8, 19 -- Date: Wed, 7 Jun 2006 23:55:43 -0700 (PDT) 8, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 19 -- Subject: Re: [Microscopy] Lead citrate staining: Thank you again and again.... 8, 19 -- To: microscopy-at-microscopy.com 8, 19 -- In-Reply-To: {p0621021bc0acd7188676-at-[10.0.1.3]} 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: text/plain; charset=iso-8859-1 8, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Thank you for your numerous answers. Lots of them show a concern about autofluorescence, but I must say that I was more concerned about the quenching by Epon and the processing steps: dehydration and embedding. I wondered if and how it could "damage" the fluorochrome.
We use Epon 812 (glycidether 100) and we just cut empty block at 300 to 500nm thickness to observe the autofluorescence with the "green" filter (our fluorochrome is Alexa488). Actually there IS some autofluorescence, but not dramatic even with a thickness of 500nm. I don't think it will perturb the observation, but it depends of course how well the alexa will sustain the processing.
An excellent remark was made, pointing out that glutaraldehyde must be avoided because it brings autofluorescence. I knew it but recalling me was not bad :-D Anyway glutar is not necessary for LM observation.
I will definitely try light fixation (4% PFA for 20 min), fast dehydration in alcohol and direct embedding in Epon.
Another concern is the localisation of the cells and cell comparments. Do some of you have an idea how well work DIC or phase contrast with cells embedded in Epon? Could I just use general staining protocols hematoxylin-eosin before embedding in Epon?
Stephane
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==============================Original Headers============================== 11, 19 -- From nizets2-at-yahoo.com Thu Jun 8 07:47:23 2006 11, 19 -- Received: from web37402.mail.mud.yahoo.com (web37402.mail.mud.yahoo.com [209.191.87.55]) 11, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k58ClNTT023147 11, 19 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 07:47:23 -0500 11, 19 -- Received: (qmail 73767 invoked by uid 60001); 8 Jun 2006 12:47:22 -0000 11, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 11, 19 -- s=s1024; d=yahoo.com; 11, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 11, 19 -- b=iV+QTQg28QV+UWQjdDyBjy4AJOhD8LlQroQUt++DI1fpwtngYP1g0BTye9C09Ku/OrUlcNGCcYXn305nvBYPPrCCh5x8htA4EZTmi/om8WXtIK2/iQjH179gPv46rnhLUg/D+9WRqfRTh7T0s4plthEYocx1ZegPqnfIsczbFfE= ; 11, 19 -- Message-ID: {20060608124722.73765.qmail-at-web37402.mail.mud.yahoo.com} 11, 19 -- Received: from [80.122.101.102] by web37402.mail.mud.yahoo.com via HTTP; Thu, 08 Jun 2006 05:47:22 PDT 11, 19 -- Date: Thu, 8 Jun 2006 05:47:22 -0700 (PDT) 11, 19 -- From: Stephane Nizet {nizets2-at-yahoo.com} 11, 19 -- Subject: Re: [Microscopy] RE: weird idea 11, 19 -- To: microscopy-at-microscopy.com 11, 19 -- In-Reply-To: {200606072041.k57KfLwR006361-at-ns.microscopy.com} 11, 19 -- MIME-Version: 1.0 11, 19 -- Content-Type: text/plain; charset=iso-8859-1 11, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
Dear Stephane, Don't use eosin...it fluoresces very intensely (in the usual "Rhodamine" wavelengths). In fact, the slide I use when teaching people how to use the confocal is a standard paraffin section stained with H&E. It never seems to bleach, and it fluoresces like mad at 488 (the connective tissue), 543, and 633.
I've looked at semi-thin resin sections with DIC...it works. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 2, 23 -- From lcgould-at-med.cornell.edu Thu Jun 8 08:28:44 2006 2, 23 -- Received: from smtp-gw2.med.cornell.edu (smtp-gw2.med.cornell.edu [157.139.3.45]) 2, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58DSgdm001635 2, 23 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 08:28:43 -0500 2, 23 -- Received: from mpx2.med.cornell.edu (pc113142-10.med.cornell.edu [140.251.11.119]) 2, 23 -- by smtp-gw2.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k58DSdim006316 2, 23 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 09:28:41 -0400 (EDT) 2, 23 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 2, 23 -- by mpx2.med.cornell.edu 2, 23 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 2, 23 -- with ESMTPA id {0J0J00KT0MRQ1450-at-mpx2.med.cornell.edu} for 2, 23 -- microscopy-at-microscopy.com; Thu, 08 Jun 2006 09:28:39 -0400 (EDT) 2, 23 -- Date: Thu, 08 Jun 2006 09:21:47 -0400 2, 23 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 2, 23 -- Subject: Re: [Microscopy] weird idea 2, 23 -- In-reply-to: {200606081248.k58Cm7jk024433-at-ns.microscopy.com} 2, 23 -- Sender: lcgould-at-med.cornell.edu 2, 23 -- To: nizets2-at-yahoo.com, Microscopy Listserver {microscopy-at-microscopy.com} 2, 23 -- Message-id: {p06230903c0add44359ef-at-[140.251.48.23]} 2, 23 -- MIME-version: 1.0 2, 23 -- Content-type: text/plain; charset=us-ascii; format=flowed 2, 23 -- References: {200606081248.k58Cm7jk024433-at-ns.microscopy.com} 2, 23 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.6.8.60432 ==============================End of - Headers==============================
I put my TEM negatives on a lightbox and photograph them with a Canon G3 digital camera. Then I convert to a positive with PhotoShop.
Geoff
aarti_harle-at-yahoo.co.in wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
==============================Original Headers============================== 9, 35 -- From mcauliff-at-umdnj.edu Thu Jun 8 08:35:24 2006 9, 35 -- Received: from zix04.umdnj.edu (zix04.UMDNJ.EDU [130.219.34.127]) 9, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58DZOFj011595 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:24 -0500 9, 35 -- Received: from zix04.umdnj.edu (ZixVPM [127.0.0.1]) 9, 35 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id C348C4BE51 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:23 -0500 (CDT) 9, 35 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) 9, 35 -- by zix04.umdnj.edu (Proprietary) with ESMTP id 05B9F4BE6A 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:21 -0500 (CDT) 9, 35 -- Received: from ([130.219.34.131]) 9, 35 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.16981519; 9, 35 -- Thu, 08 Jun 2006 09:35:01 -0400 9, 35 -- Received: from conversion-daemon.Polaris.umdnj.edu by Polaris.umdnj.edu 9, 35 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) 9, 35 -- id {0J0J00501N192Q-at-Polaris.umdnj.edu} (original mail from mcauliff-at-umdnj.edu) 9, 35 -- for microscopy-at-msa.microscopy.com; Thu, 08 Jun 2006 09:35:01 -0400 (EDT) 9, 35 -- Received: from [127.0.0.1] ([10.138.2.123]) 9, 35 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 9, 35 -- 2004)) with ESMTP id {0J0J005AJN211E-at-Polaris.umdnj.edu} ; Thu, 9, 35 -- 08 Jun 2006 09:34:50 -0400 (EDT) 9, 35 -- Date: Thu, 08 Jun 2006 09:35:48 -0400 9, 35 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} 9, 35 -- Subject: Re: [Microscopy] Scanner 9, 35 -- In-reply-to: {200606081144.k58BiOek013698-at-ns.microscopy.com} 9, 35 -- To: aarti_harle-at-yahoo.co.in, 9, 35 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} 9, 35 -- Message-id: {448827B4.5000709-at-umdnj.edu} 9, 35 -- MIME-version: 1.0 9, 35 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 9, 35 -- Content-transfer-encoding: 7BIT 9, 35 -- X-Accept-Language: en-us, en 9, 35 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) 9, 35 -- Gecko/20040804 Netscape/7.2 (ax) 9, 35 -- References: {200606081144.k58BiOek013698-at-ns.microscopy.com} ==============================End of - Headers==============================
I'll send a pdf of a short article on scanners I wrote for Microscopy Today (May 2006). My personal favourite scanner is the Epson V750 Pro (£600) and the Cheaper sister the Epson V700 (£400). The older model Epson 4990 Photo is not quite as good but cheaper at £280. All these scan film up to full A4 size, and have an easy to use twain interface. They also do reflective scans for paper and photographs. My favourite scanner review site for pro-sumer (i.e. cheap) scanners is http://www.photo-i.co.uk.
As mentioned you can use a camera and stand to copy film (used to be called an epidiascope in my Quantimet image analyser days) - and you can get pretty good optical microscope images just by resting a compact digital camera lens against the eyepiece. These days though I think for film, 6,400 dpi pro-sumer flatbed scanners do it better, more conveniently and relatively cheaply.
For image analysis there's lot to choose from, e.g. Improvision OpenLabs and Velocity (http://www.improvision.co.uk) ImageProPlus (http://www.mediacy.com) Kinetic Imaging (now http://www.andor.com/products) MetaMorph (http://www.moleculardevices.com/pages/software/metamorph.html)
These packages are expensive though (typically £3k+). They all deal with scanned tif images (and much more).
However if you don't have £3,000 for the above software, have at look at the freeware ImageJ (for the PC) at http://rsb.info.nih.gov/ij - you often need some of the plug-ins as well (there is also the sister program NIHImage for Apple fans at http://rsb.info.nih.gov/nih-image). Not always an easy program to get to grips with quickly (like most image analysis software), but quite powerful with plug-ins. For simple image manipulation, Photoshop CS2 is the obvious choice - otherwise the cut-down Photoshop Elements is pretty good and is supplied free with the above scanners (as a slightly older version).
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
I wonder if the autofluorescence might be due primarily to the MNA (aka NMA) in the usual Luft formulation?
When i found that NMA caused the cured resin to react horribly with MnO4 section stain --- (useful because the Mn04-Pb staining sequence gives much more contrast than any other stain we've ever tried) --- I learned to avoid MNA and just empirically readjusted DDSA and Epon ratios empirically for a firm enough cured resin, needing no MNA, yet compatible with MnO4 staining. However, I soon abandoned Epon and Spurr of all kinds in favor of an Araldite 506-DDSA-DER 736 mixture (10:15:2 gm by weight) because it sections thinner and accepts Mn-Pb staining with less graininess. So that Araldite mix is what i will try soon with Alexa 488-phalloidin.
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-- -mike reedy-
************************ Michael K. Reedy, M.D. Duke Univ. Med. Center Dept. Cell Biology, Box 3011 (for U.S. Mail) 458 Alex Sands Bldg, Research Dr (courier) Durham, NC 27710
Hi Mike, Interesting thought about the NMA, and your Araldite recipe looks worth a try. I had dropped the DMAE in Spurr's to 1/2 and then to 1/4 of the original recipe, thinking that it might generate radicals reacting with the fluorophores. The result was slightly reduced autofluorescence after a 3 day cure time at 60 C. No apparent effect on the fluorophores themselves, but didn't spend any time measuring.
FYI, alcohols will cause phalloidin to dissociate from the actin. Maybe it will survive a higher alcohol, propyl or butyl, but an actin antibody would be a better choice if needing to dehydrate the sample.
Regards, Glen
Glen MacDonald Core for Communication Research Virginia Merrill Bloedel Hearing Research Center Box 357923 University of Washington Seattle, WA 98195-7923 USA (206) 616-4156 glenmac-at-u.washington.edu
************************************************************************ ****** The box said "Requires Windows 95 or better", so I bought a Macintosh. ************************************************************************ ******
On Jun 8, 2006, at 9:17 AM, Mike Reedy wrote:
} Wonderful weird idea! } } I wonder if the autofluorescence might be due primarily to the MNA } (aka NMA) in the usual Luft formulation? } } When i found that NMA caused the cured resin to react horribly with } MnO4 section stain --- (useful because the Mn04-Pb staining } sequence gives much more contrast than any other stain we've ever } tried) --- I learned to avoid MNA and just empirically readjusted } DDSA and Epon ratios empirically for a firm enough cured resin, } needing no MNA, yet compatible with MnO4 staining. However, I soon } abandoned Epon and Spurr of all kinds in favor of an Araldite 506- } DDSA-DER 736 mixture (10:15:2 gm by weight) because it sections } thinner and accepts Mn-Pb staining with less graininess. So that } Araldite mix is what i will try soon with Alexa 488-phalloidin. } } -mike reedy- } } --------------------------------------------------------------------- } } ------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } --------------------------------------------------------------------- } } ------- } } } } Dear without an I, } } Success will depend upon the fluorophore. FITC and TRITC will lose } } their fluorescence. Alexa fluor dyes and cyanine dyes will work } } well. Actually, I think with reduced photobleaching. } } Autofluorescence will depend upon imaging modality. Spurr's looks } } great under the confocal, but the autolfuorescence creates an } } impossible haze with epi-fluorescence on thick samples. } } Deconvolution will clean up the haze. 3 micron sections looked very } } good. the new formulation of spurr's is too brittle so I'm going to } } try Eponate 812 or something similar for my next round of embedded } } fluorescence. I tried Histo-Resin and found the autofluorescence was } } noticeable under confocal, but didn't try LR-White. } } } } Hardie, MacDonald, Rubel, Brain Res. 1000:200-210, 2003 } } } } Regards, } } Glen } } On Jun 7, 2006, at 10:49 AM, drk-at-SHCC.org wrote: } } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } -- } } } ------ } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } America } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } } MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/ } } } FAQ.html } } } } } } -------------------------------------------------------------------- } } } -- } } } ------ } } } } } } Regarding the idea of localizing fluorescence label in epon, I } } } would suggest } } } that you not use Epon...there are autoflourescence problems (at } } } least in our } } } hands, using Spurrs resin). However, I think you may be happy } } } using LR } } } White, polymerized at 60C. We've done some similar work here and } } } have been } } } very impressed with the results. } } } } } } Good luck, } } } } } } Doug } } } } } } Douglas R. Keene } } } Assistant Investigator } } } Micro-Imaging Center } } } Shriners Hospital for Children } } } 3102 S.W. Sam Jackson Park Road } } } Portland, Oregon 97239 } } } 503-221-3434 } } } drk-at-shcc.org } } } -----Original Message----- } } } X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com] } } } Sent: Wednesday, June 07, 2006 1:10 AM } } } To: drk-at-SHCC.org } } } Subject: [Microscopy] weird idea: fluorescence in Epon } } } } } } } } } } } } } } } } } } -------------------------------------------------------------------- } } } -- } } } ------ } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } } } America } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } } } MicroscopyListserver } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/ } } } FAQ.html } } } } } } -------------------------------------------------------------------- } } } -- } } } ------ } } } } } } Dear listers, } } } } } } Here is another weird idea from me :-D } } } I would like to localize a fluorescent molecule in the } } } transversal plane of a cell monolayer(not from above). } } } We have no material for the preparation of } } } histological sections, but we have all necessary for } } } TEM. I wonder if I could not embed the monolayer in } } } Epon, then cut transversal semi-thin sections and } } } observe in epifluroscence. } } } Would Epon hinder fluorescence? } } } Could it be done in another resin than Epon? } } } } } } Regards, } } } } } } StÈphane } } }
==============================Original Headers============================== 9, 25 -- From glenmac-at-u.washington.edu Thu Jun 8 11:55:26 2006 9, 25 -- Received: from mxout1.cac.washington.edu (mxout1.cac.washington.edu [140.142.32.134]) 9, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58GtQZE014722 9, 25 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 11:55:26 -0500 9, 25 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.33.9]) 9, 25 -- by mxout1.cac.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k58GtPsV019189 9, 25 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 09:55:25 -0700 9, 25 -- X-Auth-Received: from [128.95.178.71] ([128.95.178.71]) 9, 25 -- (authenticated authid=glenmac) 9, 25 -- by smtp.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k58GtMgj017005 9, 25 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 9, 25 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 09:55:25 -0700 9, 25 -- Mime-Version: 1.0 (Apple Message framework v750) 9, 25 -- In-Reply-To: {p06210229c0adfa70cf1a-at-[10.0.1.3]} 9, 25 -- References: {200606072039.k57KdsxX003708-at-ns.microscopy.com} {p06210229c0adfa70cf1a-at-[10.0.1.3]} 9, 25 -- Content-Type: text/plain; charset=ISO-8859-1; delsp=yes; format=flowed 9, 25 -- Message-Id: {208F7BAE-D0C3-4CBF-B33A-BBFD3BDF8880-at-u.washington.edu} 9, 25 -- From: Glen MacDonald {glenmac-at-u.washington.edu} 9, 25 -- Subject: RE: weird idea 9, 25 -- Date: Thu, 8 Jun 2006 09:55:17 -0700 9, 25 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 9, 25 -- X-Mailer: Apple Mail (2.750) 9, 25 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_NAME_BODY 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __FRAUD_419_FUNWORDS 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0' 9, 25 -- Content-Transfer-Encoding: 8bit 9, 25 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k58GtQZE014722 ==============================End of - Headers==============================
Shrunali Could I suggest you check out the Microscopy List Server Archive? There has been extensive and intelligent discussion about this subject quite frequently over the last couple of years
Richard Harris Laboratory Supervisor, Imaging and Analysis Department of Biology, University of Western Ontario, London Ontario, CANADA. N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
-----Original Message----- X-from: aarti_harle-at-yahoo.co.in [mailto:aarti_harle-at-yahoo.co.in] Sent: June 8, 2006 7:48 AM To: rjharris-at-uwo.ca
Hello,
Can anyone suggest good scanner to scan TEM negatives and prints and and latest Imange Analysis software.
Regards Shrunali Kulkarni Scientist IMTech
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 4, 18 -- From aarti_harle-at-yahoo.co.in Thu Jun 8 06:42:41 2006 4, 18 -- Received: from web8324.mail.in.yahoo.com (web8324.mail.in.yahoo.com [202.43.219.162]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k58BgdP1011677 4, 18 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 06:42:40 -0500 4, 18 -- Received: (qmail 80614 invoked by uid 60001); 8 Jun 2006 11:42:38 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 18 -- s=s1024; d=yahoo.co.in; 4, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content -Transfer-Encoding; 4, 18 -- b=4O7v0G9DukweatO+2R8G5gg8q2vMGakVzH/e8sNmdfB5lHYsou5keT5GIf5MCJ3QXABL+p6TYN 2J8ErZVja7YfTq2GEVJSWlBmkVhK7a8GWj2EHuUnSD/dSKfIhsaVfnZmLPKTxxwkjJsKthJ4k7ai d29uMqt8F5DjHJQgxz+8E= ; 4, 18 -- Message-ID: {20060608114238.80612.qmail-at-web8324.mail.in.yahoo.com} 4, 18 -- Received: from [203.197.210.214] by web8324.mail.in.yahoo.com via HTTP; Thu, 08 Jun 2006 04:42:38 PDT 4, 18 -- Date: Thu, 8 Jun 2006 04:42:38 -0700 (PDT) 4, 18 -- From: Aarti Harle {aarti_harle-at-yahoo.co.in} 4, 18 --
You can purchase many different scanners for prices under $500 that will do the job for routine negatives. What is critical is that they are capable of both reflective and transmittal illumination. For negatives you need to be able to transmit the light through the negative while normal scanning of documents utilizes reflective light.
We normally scan routine negatives at 600dpi. However, if we need to enlarge the image substantially, we will often scan at 1200dpi or higher.
Debby
Debby Sherman, Director Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
} From: {mcauliff-at-umdnj.edu} } Reply-To: {mcauliff-at-umdnj.edu} } Date: Thu, 8 Jun 2006 08:37:52 -0500 } To: {dsherman-at-purdue.edu} } Subject: [Microscopy] Re: Scanner } } } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I put my TEM negatives on a lightbox and photograph them with a Canon G3 } digital camera. Then I convert to a positive with PhotoShop. } } Geoff } } } aarti_harle-at-yahoo.co.in wrote: } } } ---------------------------------------------------------------------------- } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } } Hello, } } } } Can anyone suggest good scanner to scan TEM negatives } } and prints and and latest Imange Analysis software. } } } } Regards } } Shrunali Kulkarni } } Scientist } } IMTech } } } } __________________________________________________ } } Do You Yahoo!? } } Tired of spam? Yahoo! Mail has the best spam protection around } } http://mail.yahoo.com } } } } } } } } } -- } -- } ********************************************** } Geoff McAuliffe, Ph.D. } Neuroscience and Cell Biology } Robert Wood Johnson Medical School } 675 Hoes Lane, Piscataway, NJ 08854 } voice: (732)-235-4583; fax: -4029 } mcauliff-at-umdnj.edu } ********************************************** } } } } ==============================Original Headers============================== } 9, 35 -- From mcauliff-at-umdnj.edu Thu Jun 8 08:35:24 2006 } 9, 35 -- Received: from zix04.umdnj.edu (zix04.UMDNJ.EDU [130.219.34.127]) } 9, 35 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id } k58DZOFj011595 } 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:24 -0500 } 9, 35 -- Received: from zix04.umdnj.edu (ZixVPM [127.0.0.1]) } 9, 35 -- by Outbound.umdnj.edu (Proprietary) with ESMTP id C348C4BE51 } 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:23 -0500 } (CDT) } 9, 35 -- Received: from umdnj.edu (imail2.UMDNJ.EDU [130.219.34.139]) } 9, 35 -- by zix04.umdnj.edu (Proprietary) with ESMTP id 05B9F4BE6A } 9, 35 -- for {microscopy-at-msa.microscopy.com} ; Thu, 8 Jun 2006 08:35:21 -0500 } (CDT) } 9, 35 -- Received: from ([130.219.34.131]) } 9, 35 -- by imail2.umdnj.edu with ESMTP id KP-BRCRF.16981519; } 9, 35 -- Thu, 08 Jun 2006 09:35:01 -0400 } 9, 35 -- Received: from conversion-daemon.Polaris.umdnj.edu by } Polaris.umdnj.edu } 9, 35 -- (iPlanet Messaging Server 5.2 HotFix 2.02 (built Oct 21 2004)) } 9, 35 -- id {0J0J00501N192Q-at-Polaris.umdnj.edu} (original mail from } mcauliff-at-umdnj.edu) } 9, 35 -- for microscopy-at-msa.microscopy.com; Thu, 08 Jun 2006 09:35:01 -0400 } (EDT) } 9, 35 -- Received: from [127.0.0.1] ([10.138.2.123]) } 9, 35 -- by Polaris.umdnj.edu (iPlanet Messaging Server 5.2 HotFix 2.02 } (built Oct 21 } 9, 35 -- 2004)) with ESMTP id {0J0J005AJN211E-at-Polaris.umdnj.edu} ; Thu, } 9, 35 -- 08 Jun 2006 09:34:50 -0400 (EDT) } 9, 35 -- Date: Thu, 08 Jun 2006 09:35:48 -0400 } 9, 35 -- From: Geoff McAuliffe {mcauliff-at-umdnj.edu} } 9, 35 -- Subject: Re: [Microscopy] Scanner } 9, 35 -- In-reply-to: {200606081144.k58BiOek013698-at-ns.microscopy.com} } 9, 35 -- To: aarti_harle-at-yahoo.co.in, } 9, 35 -- MicroscopyListserver {microscopy-at-msa.microscopy.com} } 9, 35 -- Message-id: {448827B4.5000709-at-umdnj.edu} } 9, 35 -- MIME-version: 1.0 } 9, 35 -- Content-type: text/plain; format=flowed; charset=ISO-8859-1 } 9, 35 -- Content-transfer-encoding: 7BIT } 9, 35 -- X-Accept-Language: en-us, en } 9, 35 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.7.2) } 9, 35 -- Gecko/20040804 Netscape/7.2 (ax) } 9, 35 -- References: {200606081144.k58BiOek013698-at-ns.microscopy.com} } ==============================End of - Headers==============================
==============================Original Headers============================== 7, 23 -- From dsherman-at-purdue.edu Thu Jun 8 13:23:12 2006 7, 23 -- Received: from 1061exfe02.adpc.purdue.edu (1061exfe02.adpc.purdue.edu [128.210.63.223]) 7, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58INBT5004486 7, 23 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 13:23:12 -0500 7, 23 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe02.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 23 -- Thu, 8 Jun 2006 14:23:11 -0400 7, 23 -- Received: from 128.210.161.94 ([128.210.161.94]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.239]) with Microsoft Exchange Server HTTP-DAV ; 7, 23 -- Thu, 8 Jun 2006 18:23:08 +0000 7, 23 -- User-Agent: Microsoft-Entourage/11.2.3.060209 7, 23 -- Date: Thu, 08 Jun 2006 14:23:07 -0400 7, 23 -- Subject: Re: [Microscopy] Re: Scanner 7, 23 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 23 -- To: {mcauliff-at-umdnj.edu} 7, 23 -- CC: "message to: MSA list" {microscopy-at-microscopy.com} 7, 23 -- Message-ID: {C0ADE34B.127D7%dsherman-at-purdue.edu} 7, 23 -- Thread-Topic: [Microscopy] Re: Scanner 7, 23 -- Thread-Index: AcaLKJa01VvqA/cbEdq0UwARJN08Mg== 7, 23 -- In-Reply-To: {200606081337.k58DbqJa015892-at-ns.microscopy.com} 7, 23 -- Mime-version: 1.0 7, 23 -- Content-type: text/plain; 7, 23 -- charset="US-ASCII" 7, 23 -- Content-transfer-encoding: 7bit 7, 23 -- X-OriginalArrivalTime: 08 Jun 2006 18:23:11.0835 (UTC) FILETIME=[99963AB0:01C68B28] ==============================End of - Headers==============================
Seeing at the Nanoscale IV, July 17-20, University of Pennsylvania, Philadelphia.
All users of SPM and metrology instruments are invited to Philadelphia next month to meet and discuss their work informally with colleagues and nanoscience pioneers from around the world!
The three-day, event-filled "Seeing at the Nanoscale" international conference includes two-and-a-half days of technical presentations, an evening poster session, and a reception at the National Constitution Center.
Technical Program: · Nanomechanical and Local Property Measurements · Visualization I: Biomolecules and Biological Processes · Visualization II: Materials and Polymer Systems · Measurements of Electrical, Optical, Magnetic, and Thermal Properties of Materials at the Nanoscale · Instrumentation: New Tools and Techniques for Nanoscience
For more information and to register online, please go to: www.veeco.com/nanoconference
____________________________ Marlene Carlyle Veeco Instruments Conference Coordinator 112 Robin Hill Road Santa Barbara, CA 93117 Tel: 805-967-1400 (ext. 2312) Fax: 805-967-7717 Email: mcarlyle-at-veeco.com ____________________________
==============================Original Headers============================== 8, 20 -- From MCarlyle-at-veeco.com Thu Jun 8 17:50:46 2006 8, 20 -- Received: from sboexch2.int.veeco.com (SBOEXCH2.VEECO.com [68.111.35.167]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k58Mojq9021045 8, 20 -- for {Microscopy-at-Microscopy.com} ; Thu, 8 Jun 2006 17:50:45 -0500 8, 20 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 20 -- content-class: urn:content-classes:message 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; 8, 20 -- charset="iso-8859-1" 8, 20 -- Subject: SPM - Seeing at the Nanoscale IV Conference 8, 20 -- Date: Thu, 8 Jun 2006 15:50:44 -0700 8, 20 -- Message-ID: {625BD6DD96DE554DBB0558FFE70E0F4201AD54D5-at-sboexch2.int.veeco.com} 8, 20 -- X-MS-Has-Attach: 8, 20 -- X-MS-TNEF-Correlator: 8, 20 -- Thread-Topic: SPM - Seeing at the Nanoscale IV Conference 8, 20 -- Thread-Index: AcaLTfmt6Y/+7q5oQh++fBJptUPOrw== 8, 20 -- From: "Marlene Carlyle" {MCarlyle-at-veeco.com} 8, 20 -- To: {Microscopy-at-Microscopy.com} 8, 20 -- Content-Transfer-Encoding: 8bit 8, 20 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k58Mojq9021045 ==============================End of - Headers==============================
Yes, this has been a recurring issue. The archives are a good source for reflection.
Also, one should scan as a transparency rather than as a negative. Then invert in PS or whatever image app you use. Maximum D value is highly desirable and this is engaged with transparencies.
gary g.
At 10:54 AM 6/8/2006, you wrote:
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==============================Original Headers============================== 9, 20 -- From gary-at-gaugler.com Thu Jun 8 18:06:35 2006 9, 20 -- Received: from qsmtp1.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 9, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k58N6ZD1031643 9, 20 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 18:06:35 -0500 9, 20 -- Received: (qmail 18405 invoked from network); 8 Jun 2006 16:06:34 -0700 9, 20 -- Received: by simscan 1.1.0 ppid: 18402, pid: 18403, t: 0.2173s 9, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 9, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 9, 20 -- by qsmtp1 with SMTP; 8 Jun 2006 16:06:34 -0700 9, 20 -- Message-Id: {7.0.1.0.2.20060608160413.023955c8-at-gaugler.com} 9, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 9, 20 -- Date: Thu, 08 Jun 2006 16:06:35 -0700 9, 20 -- To: rjharris-at-uwo.ca 9, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 9, 20 -- Subject: Re: [Microscopy] RE: Scanner 9, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 9, 20 -- In-Reply-To: {200606081754.k58Hslnn029206-at-ns.microscopy.com} 9, 20 -- References: {200606081754.k58Hslnn029206-at-ns.microscopy.com} 9, 20 -- Mime-Version: 1.0 9, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both fisher.phyllis-at-mayo.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Question: I am considering adding a cathode luminescence tip to an old Centaurus backscatter detector on an Hitachi 4700 FESEM. Does anyone have experience with this tip and/or suggestions or comments regarding such an installation?
Dear Group, I am looking for someone who is going to participate the meeting on Microscopy and Microanalysis 2006 in Chicago and wants to share a room (hotel, hostel or guest house). Please inform me offline!
Grzegorz Tylko Department of Cytology and Histology Institute of Zoology Jagiellonian Univeristy Ingardena 6, 30-060 Krakow Poland tel.: +48 12 663 24 25 fax: +48 12 634 49 51 mobile: +48 698 55 55 85 e-mail: tylko-at-zuk.iz.uj.edu.pl
==============================Original Headers============================== 4, 37 -- From tylko-at-zuk.iz.uj.edu.pl Fri Jun 9 01:45:34 2006 4, 37 -- Received: from theta.uoks.uj.edu.pl (theta.uoks.uj.edu.pl [149.156.89.30]) 4, 37 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k596jXRs028765 4, 37 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 01:45:33 -0500 4, 37 -- Received: from zuk.iz.uj.edu.pl (zuk.iz.uj.edu.pl [149.156.72.18]) 4, 37 -- by theta.uoks.uj.edu.pl (8.13.6/8.13.6) with ESMTP id k596jUlp002534 4, 37 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 08:45:31 +0200 (CEST) 4, 37 -- Received: from ZUK/SpoolDir by zuk.iz.uj.edu.pl (Mercury 1.48); 4, 37 -- 9 Jun 06 08:45:31 +0100 4, 37 -- Received: from SpoolDir by ZUK (Mercury 1.48); 9 Jun 06 08:45:08 +0100 4, 37 -- Received: from gt (149.156.77.111) by zuk.iz.uj.edu.pl (Mercury 1.48); 4, 37 -- 9 Jun 06 08:45:04 +0100 4, 37 -- Message-ID: {003301c68b90$3d231a60$6f4d9c95-at-gt} 4, 37 -- From: "Grzegorz Tylko" {tylko-at-zuk.iz.uj.edu.pl} 4, 37 -- To: {microscopy-at-microscopy.com} 4, 37 -- Subject: MM2006-accommodation 4, 37 -- Date: Fri, 9 Jun 2006 08:45:04 +0200 4, 37 -- MIME-Version: 1.0 4, 37 -- Content-Type: text/plain; 4, 37 -- format=flowed; 4, 37 -- charset="iso-8859-2"; 4, 37 -- reply-type=original 4, 37 -- Content-Transfer-Encoding: 7bit 4, 37 -- X-Priority: 3 4, 37 -- X-MSMail-Priority: Normal 4, 37 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 4, 37 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 4, 37 -- X-Antivirus: Dr.Web (R) for Mail Servers on pandora host 4, 37 -- X-Antivirus-Code: 100000 4, 37 -- X-Spam-Flag: NO 4, 37 -- X-Spam-Status: score=-5.9 required=5.0 4, 37 -- X-Spam-Report: 4, 37 -- * -3.3 ALL_TRUSTED Passed through trusted hosts only via SMTP 4, 37 -- * -2.6 BAYES_00 BODY: Bayesian spam probability is 0 to 1% 4, 37 -- * [score: 0.0000] 4, 37 -- X-Spam-Checker-Version: SpamAssassin 3.0.5 (2005-11-28) on 4, 37 -- pandora.uoks.uj.edu.pl ==============================End of - Headers==============================
The only problem with archived scanner info is that it goes out of date the minute a new scanner hits the streets. I bought a £250 Canon 9550F at home [Xmas 2005] under the mistaken view that consumer flatbeds can't get any better than this for the price - I was very wrong (and annoyingly it was my money). Going back 5 years a scanner that produced images like the 9950F would have cost £4,000.
And I remember paying £5,000 for an office 386 computer with 8Mb of RAM and a 120Mb hard drive back in 1989 - I wouldn't recommend it as particularly good value these days. It did allow me to ditch the expensive IBM central mainframe and run Fortran FTN77 independently on my PC though.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: rjharris-at-uwo.ca [mailto:rjharris-at-uwo.ca] Sent: 08 June 2006 18:57 To: keith.morris-at-ucl.ac.uk
Shrunali Could I suggest you check out the Microscopy List Server Archive? There has been extensive and intelligent discussion about this subject quite frequently over the last couple of years
Richard Harris Laboratory Supervisor, Imaging and Analysis Department of Biology, University of Western Ontario, London Ontario, CANADA. N6A 5B7 Ph. 519-661-2111 ext. 86780 Fax 519-661-3935
-----Original Message----- X-from: aarti_harle-at-yahoo.co.in [mailto:aarti_harle-at-yahoo.co.in] Sent: June 8, 2006 7:48 AM To: rjharris-at-uwo.ca
Hello,
Can anyone suggest good scanner to scan TEM negatives and prints and and latest Imange Analysis software.
Regards Shrunali Kulkarni Scientist IMTech
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 4, 18 -- From aarti_harle-at-yahoo.co.in Thu Jun 8 06:42:41 2006 4, 18 -- Received: from web8324.mail.in.yahoo.com (web8324.mail.in.yahoo.com [202.43.219.162]) 4, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k58BgdP1011677 4, 18 -- for {microscopy-at-microscopy.com} ; Thu, 8 Jun 2006 06:42:40 -0500 4, 18 -- Received: (qmail 80614 invoked by uid 60001); 8 Jun 2006 11:42:38 -0000 4, 18 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 4, 18 -- s=s1024; d=yahoo.co.in; 4, 18 -- h=Message-ID:Received:Date:From:Subject:To:MIME-Version:Content-Type:Content -Transfer-Encoding; 4, 18 -- b=4O7v0G9DukweatO+2R8G5gg8q2vMGakVzH/e8sNmdfB5lHYsou5keT5GIf5MCJ3QXABL+p6TYN 2J8ErZVja7YfTq2GEVJSWlBmkVhK7a8GWj2EHuUnSD/dSKfIhsaVfnZmLPKTxxwkjJsKthJ4k7ai d29uMqt8F5DjHJQgxz+8E= ; 4, 18 -- Message-ID: {20060608114238.80612.qmail-at-web8324.mail.in.yahoo.com} 4, 18 -- Received: from [203.197.210.214] by web8324.mail.in.yahoo.com via HTTP; Thu, 08 Jun 2006 04:42:38 PDT 4, 18 -- Date: Thu, 8 Jun 2006 04:42:38 -0700 (PDT) 4, 18 -- From: Aarti Harle {aarti_harle-at-yahoo.co.in} 4, 18 --
Hi all,
I have been attempting to prepare cross sectional TEM samples of anodic tantala (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an approximate thickness of 200um. The oxide film is grown anodically on the Ta using electrochemistry.
In the literature, I have seen cross sectional TEM images of anodic tantala grown on sputter deposited tantalum; an aluminum or silicon substrate is typically used. I am curious if anyone has had success in preparing anodic Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)
My main problem seems to be with de-lamination of the cross section sandwich. Here is my attempted procedure:
I first bound two faces of my sample together and placed the sandwich in a press while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich frequently breaks apart when I begin polishing. To address this problem, I encased my sandwich in a brass tube and fitted the tube with supporting brass shims, all held in place with epoxy. Using this technique, I was able to thin some parts of the specimen to 20-50 um, but the epoxy was no longer present at this point. When I attempted to ion-mill, the sandwiches splintered apart, and any of the oxide that may have been surviving was obliterated in the ion mill.
Any suggestions or advice would be greatly appreciated.
Thank You,
Jennifer Ray Sloppy Materials Science & Engineering Department Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752 9, 17 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 11:45:10 -0500 9, 17 -- Received: (from webmail-at-localhost) 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431; 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Message-Id: {200606091645.MAA01431-at-webmail15.cac.psu.edu} 9, 17 -- To: microscopy-at-microscopy.com 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep 9, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} 9, 17 -- X-Mailer: Penn State WebMail 9, 17 -- X-Sender: jds451 9, 17 -- X-Originating-IP: 128.118.37.54 9, 17 -- MIME-Version: 1.0 9, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
Jennifer Ray Sloppy wrote: ==================================================== I have been attempting to prepare cross sectional TEM samples of anodic tantala (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an approximate thickness of 200um. The oxide film is grown anodically on the Ta using electrochemistry.
In the literature, I have seen cross sectional TEM images of anodic tantala grown on sputter deposited tantalum; an aluminum or silicon substrate is typically used. I am curious if anyone has had success in preparing anodic Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)
My main problem seems to be with de-lamination of the cross section sandwich. Here is my attempted procedure:
I first bound two faces of my sample together and placed the sandwich in a press while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich frequently breaks apart when I begin polishing. To address this problem, I encased my sandwich in a brass tube and fitted the tube with supporting brass shims, all held in place with epoxy. Using this technique, I was able to thin some parts of the specimen to 20-50 um, but the epoxy was no longer present at this point. When I attempted to ion-mill, the sandwiches splintered apart, and any of the oxide that may have been surviving was obliterated in the ion mill.
Any suggestions or advice would be greatly appreciated. ============================================================ Some number of years ago, when our firm was first getting active in surface analysis and the need for calibrating etch rates became apparent, and the "standard" at the time for calibrating etching rates was to etch through an anodic film grown on a thin tantalum foil of known thickness, we found this could be cross-sectioned by diamond knife ultramicrotomy. Other than possessing a lot of experience with the ultramicrotomy of "difficult and hard" samples, we vacuum embedded the sample with our own SPI-Pon™ 812 epoxy embedding resin, and did the thin sectioning with a 45 deg angle SPI Supplies® Brand Materials Science Diamond Knife but with a fairly ordinary (e.g. not very new) ultramicrotome. Separation of the anodized layer from the embedding resin could be minimized by using a vacuum impregnation. We were not always successful in keeping the anodized layer/substrate tantalum interface intact. We could get less separation by using a 35 deg knife but this wore out the knife very quickly and since we did not mind some separation, we used mainly a 45 deg. knife.
But one could get good TEM views of the ultramicrotome-produced cross-section for making (anodized layer) thickness measurements and high resolution images showing the morphology and fine structure of the anodized layer.
Disclaimer: SPI Supplies offers both the above-mentioned embedding resin and diamond knife and we also offer, through Structure Probe, Inc. this kind of a laboratory service for others.
Chuck
================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: http://www.2spi.com ############################ ==================================================
==============================Original Headers============================== 16, 24 -- From cgarber-at-2spi.com Fri Jun 9 13:43:39 2006 16, 24 -- Received: from diskless5.axs2000.net (mail.axs2000.net [209.120.196.43]) 16, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59IhdBg009769 16, 24 -- for {microscopy-at-msa.microscopy.com} ; Fri, 9 Jun 2006 13:43:39 -0500 16, 24 -- Received: from ibm1x23g2abfyg ([68.246.87.223]) 16, 24 -- by diskless5.axs2000.net (8.12.11/8.12.11) with SMTP id k59IhXNu008521; 16, 24 -- Fri, 9 Jun 2006 14:43:36 -0400 16, 24 -- X-IDV-FirstRcvd: [68.246.87.223] 16, 24 -- X-IDV-HELO: ibm1x23g2abfyg 16, 24 -- Message-ID: {00b401c68bf4$9b4db500$df57f644-at-ibm1x23g2abfyg} 16, 24 -- Reply-To: "Garber, Charles A." {cgarber-at-2spi.com} 16, 24 -- From: "Garber, Charles A." {cgarber-at-2spi.com} 16, 24 -- To: "Microscopy Listserver" {microscopy-at-msa.microscopy.com} 16, 24 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep 16, 24 -- Date: Fri, 9 Jun 2006 14:39:43 -0400 16, 24 -- MIME-Version: 1.0 16, 24 -- Content-Type: text/plain; 16, 24 -- charset="Windows-1252" 16, 24 -- X-Priority: 3 16, 24 -- X-MSMail-Priority: Normal 16, 24 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 16, 24 -- X-MIMEOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 16, 24 -- Content-Transfer-Encoding: 8bit 16, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k59IhdBg009769 ==============================End of - Headers==============================
I have a colleague who wants to do in situ hybridizations on sponge larvae (yes, sponges are animals and they have larval forms!). The larvae are less than a millimeter long and quite delicate.
She tried fixing some in 4% paraformaldehyde in sea water, and the larvae immediately exploded. She also tried Carnoy's fix, and 4% PFA in Millonig's phosphate buffer, and the same thing happened.
The only thing I could think of to suggest was dropping to 1 or 2% PFA in sea water. I've seen references to using Bouins for light microscopy or osmium tetroxide for TEM, but I don't think either of these approaches would be good for in situs.
Any other suggestions?
Gary P. Radice Department of Biology 804-289-8107 (voice) University of Richmond 804-289-8233 (FAX) Richmond VA 23173
==============================Original Headers============================== 9, 21 -- From gradice-at-richmond.edu Fri Jun 9 13:48:42 2006 9, 21 -- Received: from monty.richmond.edu (monty.richmond.edu [141.166.24.29]) 9, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59Imfs9015519 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 13:48:42 -0500 9, 21 -- Received: from rayon.richmond.edu (rayon.richmond.edu [141.166.30.10]) 9, 21 -- by monty.richmond.edu (8.11.6/8.11.6) with ESMTP id k59Imf119051 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 14:48:41 -0400 9, 21 -- Received: from [141.166.185.175] (ffa185175.richmond.edu [141.166.185.175]) 9, 21 -- by rayon.richmond.edu (8.12.11.20060308/8.12.11) with ESMTP id k59ImfT5023124 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 14:48:41 -0400 9, 21 -- Mime-Version: 1.0 (Apple Message framework v750) 9, 21 -- Content-Transfer-Encoding: 7bit 9, 21 -- Message-Id: {063B77D0-FAF3-4B5D-AB90-04354229FE6A-at-richmond.edu} 9, 21 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 9, 21 -- To: microscopy-at-microscopy.com 9, 21 -- From: Gary Radice {gradice-at-richmond.edu} 9, 21 -- Subject: fixing sponge larvae for in situ hybridization 9, 21 -- Date: Fri, 9 Jun 2006 14:47:52 -0400 9, 21 -- X-Mailer: Apple Mail (2.750) 9, 21 -- X-Spam-Detail: -1.44 ALL_TRUSTED 9, 21 -- X-Scanned-By: MIMEDefang 2.49 on 141.166.30.10 ==============================End of - Headers==============================
There is a trick that I pulled with MoS2 films on Si in which I had similar problems as you are having. What I did was to first deposit the film through a gridded mask (I think that it was 400 mesh, but can't remember for sure.) Then I prepared the samples as normal. What happens is that you have enough Si-epoxy-Si areas when the samples are put together to hold the sample stack together. It worked quite well. For you, you will have to make a mask to prevent the anodic Ta2O5 from forming and then remove your mask prior to making your cross section.
I also have another solution that might work for you. These are Stainless steel rods that were electrospark cut with slits in them. You must precisely cut the width of two strips of material and remove the burrs and put them together between the rods. This gives you more support with less glue interfaces. If you send me your address, I can send you some 3mm tubes and some rods to try out.
The third method that might work for you is the Technoorg-Linda method. This is illustrated in the Arpad Barna article: Barna, A. (1992) "Topographic kinetics and practice of low-angle ion beam thinning", Specimen Preparation for TEM of Materials. III, Mat. Res. Soc. Symp. Proc. Vol. 254. R. Anderson, B. Tracy, and J. Bravman, eds. pp. 3–22. You can also check the following articles: "Preparation of Cross-Sectional TEM Samples for Low-Angle Ion Milling", J.P. MCCAFFREY AND A. BARNA, MICROSCOPY RESEARCH AND TECHNIQUE 36:362–367 (1997). I can also send you a copy of a video that shows how to do it. Basically, two pieces of the sample are cut down to precise sizes and put in a special titanium grid face-to-face. Then the grid is manipulated with tools to clamp the samples in place tightly. The assembly is then put into an epoxy. You can use M-bond 610, but T-L recommends a special Araldit mixture with carbon that avoids the differential sputter removal problem with non-filled epoxies. The sample is then ground down with 600 grit paper to a thickness of about 30-50 µm. You polish with 1 µm paste to finish. It is then ion milled at a high voltage and a low angle, say about 5 to 7 degrees with full rotation for say about an hour depending on the sputter rate. This will give a dimple in the epoxy joint. Then go to a very low angle, ~2 degrees, on one side of the sample and mill until the topography is pushed to one side of the sample. Not all commercially available ion mills are capable of going to such a low angle. You must use a holder that is capable of it. Some mills have holders that are more massive that allow the heat to be dissipated more readily. Use the holder that is made to be milled from one side and at very low angles. You will see the topography of the sample pushed to the opposite side of the sample from where the gun is. Turn the sample over and mill in such a way that the ion direction would appear to oppose the direction from the first side milling. When you are done, the topography will again be pushed away from the ion gun and it will be on the opposite side of the sample of the glue joint than where it is on the first side. Finish the sample using the same angles, but low energy. Unless you have a special low energy gun such as the one available from Technoorg-Linda, most commercially available guns are limited to about 3 kV because they do not efficiently ionize the gas and the beam spread. The low energy guns available from T-L will go as low as 100 eV.
If you have any questions, please give me a call.
-Scott
Scott D. Walck, Ph.D. Technical Director South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673
US Toll Free: 1-800-728-2233 Tel: (949) 492-2600 Fax: (949) 492-1499
www.southbaytech.com walck-at-southbaytech.com
-----Original Message----- X-from: jds451-at-psu.edu [mailto:jds451-at-psu.edu] Sent: Friday, June 09, 2006 9:49 AM To: Walck-at-SouthBayTech.com
Hi all,
I have been attempting to prepare cross sectional TEM samples of anodic tantala (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an approximate thickness of 200um. The oxide film is grown anodically on the Ta using electrochemistry.
In the literature, I have seen cross sectional TEM images of anodic tantala grown on sputter deposited tantalum; an aluminum or silicon substrate is typically used. I am curious if anyone has had success in preparing anodic Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)
My main problem seems to be with de-lamination of the cross section sandwich. Here is my attempted procedure:
I first bound two faces of my sample together and placed the sandwich in a press while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich frequently breaks apart when I begin polishing. To address this problem, I encased my sandwich in a brass tube and fitted the tube with supporting brass shims, all held in place with epoxy. Using this technique, I was able to thin some parts of the specimen to 20-50 um, but the epoxy was no longer present at this point. When I attempted to ion-mill, the sandwiches splintered apart, and any of the oxide that may have been surviving was obliterated in the ion mill.
Any suggestions or advice would be greatly appreciated.
Thank You,
Jennifer Ray Sloppy Materials Science & Engineering Department Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752 9, 17 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 11:45:10 -0500 9, 17 -- Received: (from webmail-at-localhost) 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431; 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Message-Id: {200606091645.MAA01431-at-webmail15.cac.psu.edu} 9, 17 -- To: microscopy-at-microscopy.com 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep 9, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} 9, 17 -- X-Mailer: Penn State WebMail 9, 17 -- X-Sender: jds451 9, 17 -- X-Originating-IP: 128.118.37.54 9, 17 -- MIME-Version: 1.0 9, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
==============================Original Headers============================== 24, 28 -- From walck-at-southbaytech.com Fri Jun 9 14:21:22 2006 24, 28 -- Received: from ylpvm29.prodigy.net (ylpvm29-ext.prodigy.net [207.115.57.60]) 24, 28 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59JLLic030645 24, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 14:21:21 -0500 24, 28 -- Received: from pimout5-ext.prodigy.net (pimout5-int.prodigy.net [207.115.4.21]) 24, 28 -- by ylpvm29.prodigy.net (8.12.10 outbound/8.12.10) with ESMTP id k59JKHVu031016 24, 28 -- for {Microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 15:20:18 -0400 24, 28 -- X-ORBL: [64.169.193.90] 24, 28 -- Received: from dynamicbl8uno3 (adsl-64-169-193-90.dsl.lsan03.pacbell.net [64.169.193.90]) 24, 28 -- by pimout5-ext.prodigy.net (8.13.6 out.dk/8.13.6) with ESMTP id k59JLG3o212652; 24, 28 -- Fri, 9 Jun 2006 15:21:16 -0400 24, 28 -- From: "Scott Walck" {walck-at-southbaytech.com} 24, 28 -- To: {jds451-at-psu.edu} 24, 28 -- Cc: {Microscopy-at-microscopy.com} 24, 28 -- Subject: RE: [Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep 24, 28 -- Date: Fri, 9 Jun 2006 12:21:21 -0700 24, 28 -- Message-ID: {002801c68bf9$e4fd4940$7801a8c0-at-dynamicbl8uno3} 24, 28 -- MIME-Version: 1.0 24, 28 -- Content-Type: text/plain; 24, 28 -- charset="iso-8859-1" 24, 28 -- X-Priority: 3 (Normal) 24, 28 -- X-MSMail-Priority: Normal 24, 28 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 24, 28 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2180 24, 28 -- Importance: Normal 24, 28 -- In-Reply-To: {200606091648.k59GmbKg003550-at-ns.microscopy.com} 24, 28 -- Content-Transfer-Encoding: 8bit 24, 28 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k59JLLic030645 ==============================End of - Headers==============================
For cross sectional samples of metal/metal oxide, a rocking motion in ion mill (or single section mode in PIPS) serves best. Align the glue line perpendicular with the ion beam, allowing the beam ONLY bombarding from the backside of the film. With such arrangement, the substrate is acting as a shield for the oxide film, protecting it from being overly milled.
Chengyu Song National Center for Electron Microscopy Lawrence Berkeley National Laboratory
jds451-at-psu.edu wrote:
} ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi all, } } I have been attempting to prepare cross sectional TEM samples of anodic tantala } (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an } approximate thickness of 200um. The oxide film is grown anodically on the Ta } using electrochemistry. } } In the literature, I have seen cross sectional TEM images of anodic tantala } grown on sputter deposited tantalum; an aluminum or silicon substrate is } typically used. I am curious if anyone has had success in preparing anodic } Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer) } } My main problem seems to be with de-lamination of the cross section sandwich. } Here is my attempted procedure: } } I first bound two faces of my sample together and placed the sandwich in a press } while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich } frequently breaks apart when I begin polishing. To address this problem, I } encased my sandwich in a brass tube and fitted the tube with supporting brass } shims, all held in place with epoxy. Using this technique, I was able to thin } some parts of the specimen to 20-50 um, but the epoxy was no longer present at } this point. When I attempted to ion-mill, the sandwiches splintered apart, and } any of the oxide that may have been surviving was obliterated in the ion mill. } } Any suggestions or advice would be greatly appreciated. } } Thank You, } } Jennifer Ray Sloppy } Materials Science & Engineering Department } Ph.D. Candidate } Pennsylvania State University } } } ==============================Original Headers============================== } 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006 } 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201]) } 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752 } 9, 17 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 11:45:10 -0500 } 9, 17 -- Received: (from webmail-at-localhost) } 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431; } 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT) } 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT) } 9, 17 -- Message-Id: {200606091645.MAA01431-at-webmail15.cac.psu.edu} } 9, 17 -- To: microscopy-at-microscopy.com } 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep } 9, 17 -- From: "Jen Sloppy" {jds451-at-psu.edu} } 9, 17 -- X-Mailer: Penn State WebMail } 9, 17 -- X-Sender: jds451 } 9, 17 -- X-Originating-IP: 128.118.37.54 } 9, 17 -- MIME-Version: 1.0 } 9, 17 -- Content-Type: text/plain } ==============================End of - Headers==============================
==============================Original Headers============================== 6, 25 -- From csong-at-lbl.gov Fri Jun 9 17:56:04 2006 6, 25 -- Received: from mta2.lbl.gov (mta2.lbl.gov [128.3.41.12]) 6, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59Mu3DI012665 6, 25 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 17:56:03 -0500 6, 25 -- Received: from mta2.lbl.gov (localhost [127.0.0.1]) 6, 25 -- by mta2.lbl.gov (8.13.6/8.13.6) with ESMTP id k59Mu2tk022358 6, 25 -- for {microscopy-at-microscopy.com} ; Fri, 9 Jun 2006 15:56:02 -0700 (PDT) 6, 25 -- Received: from lbl.gov (apple-0-5-2-e0-6a-f3.dhcp.lbl.gov [131.243.3.239]) 6, 25 -- by mta2.lbl.gov (8.13.6/8.13.6) with ESMTP id k59Mu1Ys022355; 6, 25 -- Fri, 9 Jun 2006 15:56:01 -0700 (PDT) 6, 25 -- Message-ID: {4489FCBD.6117147D-at-lbl.gov} 6, 25 -- Date: Fri, 09 Jun 2006 15:57:49 -0700 6, 25 -- From: chengyu song {csong-at-lbl.gov} 6, 25 -- Reply-To: csong-at-lbl.gov 6, 25 -- Organization: lbl 6, 25 -- X-Mailer: Mozilla 4.73C-CCK-MCD LBNL V4.73 Build 2 (Macintosh; U; PPC) 6, 25 -- X-Accept-Language: en,pdf 6, 25 -- MIME-Version: 1.0 6, 25 -- To: jds451-at-psu.edu, microscopy-at-microscopy.com 6, 25 -- Subject: Re: [Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep 6, 25 -- References: {200606091648.k59GmVj4003324-at-ns.microscopy.com} 6, 25 -- Content-Type: text/plain; charset=gb2312; x-mac-type="54455854"; x-mac-creator="4D4F5353" 6, 25 -- Content-Transfer-Encoding: 7bit 6, 25 -- X-Virus-Scanned: ClamAV 0.88.2/1524/Fri Jun 9 14:28:03 2006 on mta2 6, 25 -- X-Virus-Status: Clean ==============================End of - Headers==============================
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Title-Subject: [Filtered] SEM - cathode luminescence detector
Question: I am considering adding a cathode luminescence tip to an old Centaurus backscatter detector on an Hitachi 4700 FESEM. Do you have any suggestions or comments on such a move?
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Email: warner.rr-at-pg.com Name: Ron Warner
Organization: Procter & Gamble Co.
Title-Subject: [Filtered] Reichert CS Auto, free to a good home
Question: We have the ancient Reichert CS Auto freeze substitution unit available to a good home. It was working when we last used it, about 3 years ago. The body shows some of its years (so does mine), but the internals should be fine. You pay the shipping.
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Email: aetmicro-at-optonline.net Name: Andrew Thelian
Organization: Nanoprobes, Inc.
Title-Subject: [Filtered] Philips EM 300 camera chamber issues
Question: Hi All :),
I had an issues with the plate chambers on my Philips EM 300... The venting valve jammed... So I naturally took it upon myself to attempt a repair... the overall repair seemed to have worked, the valve knob no longer jams or sticks like it had...
my problem now is...
when turning the knob(clockwise) to 10/4 o clock... the vacuum pump initiates like it is suppose to... i then press the knob in creating a seal(i wait on the vacuum light sequence)... then the last step is to finish the circuit and leave the knob at its resting position at 9/3 o clock... the rough pump shuts off as i attempt to complete the circuit...
I really doubt that PFA in itseld could make a structure explode. Your colleague should look at the osmoticity of the medium used for the fixation. I don't know sponges very well, but there is a chance that the intracellular osmoticity is regulated and is not the same as the extracellular medium (sea water). If the fixative stops the ion pumps present in the membrane, it may be possible that the high salt concentration of sea water provokes an osmotic shock (even though in this case one should expect a shrinkage of the cell, but you never know). If the intracellular osmotocity of sponge cells is unknown, he'd better try fixation media with different osmotic forces.
Stephane
--- gradice-at-richmond.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The } Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.microscopy.com/MicroscopyListserver } On-Line Help } http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } I have a colleague who wants to do in situ } hybridizations on sponge } larvae (yes, sponges are animals and they have } larval forms!). The } larvae are less than a millimeter long and quite } delicate. } } } She tried fixing some in 4% paraformaldehyde in sea } water, and the } larvae immediately exploded. She also tried Carnoy's } fix, and 4% PFA } in Millonig's } phosphate buffer, and the same thing happened. } } The only thing I could think of to suggest was } dropping to 1 or 2% } PFA in sea water. I've seen references to using } Bouins for light } microscopy or osmium tetroxide for TEM, but I don't } think either of } these approaches would be good for in situs. } } } Any other suggestions? } } } Gary P. Radice } Department of Biology 804-289-8107 (voice) } University of Richmond 804-289-8233 (FAX) } Richmond VA 23173 } } } ==============================Original } Headers============================== } 9, 21 -- From gradice-at-richmond.edu Fri Jun 9 } 13:48:42 2006 } 9, 21 -- Received: from monty.richmond.edu } (monty.richmond.edu [141.166.24.29]) } 9, 21 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k59Imfs9015519 } 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 } Jun 2006 13:48:42 -0500 } 9, 21 -- Received: from rayon.richmond.edu } (rayon.richmond.edu [141.166.30.10]) } 9, 21 -- by monty.richmond.edu (8.11.6/8.11.6) with } ESMTP id k59Imf119051 } 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 } Jun 2006 14:48:41 -0400 } 9, 21 -- Received: from [141.166.185.175] } (ffa185175.richmond.edu [141.166.185.175]) } 9, 21 -- by rayon.richmond.edu } (8.12.11.20060308/8.12.11) with ESMTP id } k59ImfT5023124 } 9, 21 -- for {microscopy-at-microscopy.com} ; Fri, 9 } Jun 2006 14:48:41 -0400 } 9, 21 -- Mime-Version: 1.0 (Apple Message framework } v750) } 9, 21 -- Content-Transfer-Encoding: 7bit } 9, 21 -- Message-Id: } {063B77D0-FAF3-4B5D-AB90-04354229FE6A-at-richmond.edu} } 9, 21 -- Content-Type: text/plain; charset=US-ASCII; } delsp=yes; format=flowed } 9, 21 -- To: microscopy-at-microscopy.com } 9, 21 -- From: Gary Radice {gradice-at-richmond.edu} } 9, 21 -- Subject: fixing sponge larvae for in situ } hybridization } 9, 21 -- Date: Fri, 9 Jun 2006 14:47:52 -0400 } 9, 21 -- X-Mailer: Apple Mail (2.750) } 9, 21 -- X-Spam-Detail: -1.44 ALL_TRUSTED } 9, 21 -- X-Scanned-By: MIMEDefang 2.49 on } 141.166.30.10 } ==============================End of - } Headers============================== }
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==============================Original Headers============================== 8, 20 -- From nizets2-at-yahoo.com Mon Jun 12 02:48:31 2006 8, 20 -- Received: from web37410.mail.mud.yahoo.com (web37410.mail.mud.yahoo.com [209.191.87.63]) 8, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5C7mVOh008885 8, 20 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 02:48:31 -0500 8, 20 -- Received: (qmail 1895 invoked by uid 60001); 12 Jun 2006 07:48:30 -0000 8, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 8, 20 -- s=s1024; d=yahoo.com; 8, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 8, 20 -- b=aQZxRVgMXGOewWw84RPkTRum1TKUECd1jKiE4vTgiSiUlxyZU6MXAexB+h8c1oO9fBMN2EDCBtGE2krNmPSuTCm4032fu7lSkwMC39aWgnfANNEbEv7I6ZUoKxkaTk14YiSdlp2eZqP8YPWj7g0CxPiCufoTJy7eFTBIm1rVQr4= ; 8, 20 -- Message-ID: {20060612074830.1893.qmail-at-web37410.mail.mud.yahoo.com} 8, 20 -- Received: from [80.122.101.102] by web37410.mail.mud.yahoo.com via HTTP; Mon, 12 Jun 2006 00:48:30 PDT 8, 20 -- Date: Mon, 12 Jun 2006 00:48:30 -0700 (PDT) 8, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 8, 20 -- Subject: Re: [Microscopy] fixing sponge larvae for in situ hybridization 8, 20 -- To: gradice-at-richmond.edu 8, 20 -- Cc: microscopy-at-microscopy.com 8, 20 -- In-Reply-To: {200606091852.k59Iqpon027420-at-ns.microscopy.com} 8, 20 -- MIME-Version: 1.0 8, 20 -- Content-Type: text/plain; charset=iso-8859-1 8, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I notice that the Epson V750 Pro flatbed scanner now has an optional fluid mounting accessory that involves a bottom sealed lift out tray (that 'prevents' fluid seepage into the scanner and is 'easy clean'). See it at:
Professional drum scanners use mounting fluid to improve film scan quality if you really need the best possible scan, and have a private income (most of us don't). If you are flatbed scanning odd specimens or really want to use oil or mounting fluid, this accessory looks very useful. I don't think the kit's generally available yet though.
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: mey-at-amnh.org [mailto:mey-at-amnh.org] Sent: 25 May 2006 20:47 To: keith.morris-at-ucl.ac.uk
Hi Mike, I have been exploring and having success using a flatbed scanner for particle analysis, but actually started out in this endeavor by scanning rocks and rock cores. We now use an Epson XL 1640 scanner, which has a focus function, essentially allowing us to get right on the surface of the samples. However, a pretty flat cut is needed to get overall excellent quality. I found that this works much better than greasing up the bed with oils, glycerin or other sticky stuff. We used a thin plastic sheet (same material as thin clear view-foils) to protect the glass from the rocks. As a side note, I too saw some strange artifacts with oils, which could (?) be due to "internal" reflections or interference patterns between the glass-plastic or micro-bubbles on or close the sample surface. My major concern was contaminating the samples with oils, especially when doing trace element work or isotope analysis for age dating purposes of the rock cores. Not that the oil itself would be the major culprit, but more the rock dust flying around everywhere in the cutting lab sticking to the greasy surfaces. Rock cores sometimes have oil on them from the drilling process and is usually washed off with sulfo. However, the porosity of the sample may contribute to deeper contamination, especially is if the sample shows cracks or micro-fractures. I had to consider this for archival purposes and potential future analysis on the same samples as new technologies emerge.
michael-at-Shaffer.net wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } We have been using a flatbed scanner for documenting rock core before } processing it for other analytical techniques. Now, we want to explore the } technique a bit further, possibly towards applied image analysis, but need } one last ingredient for better quality scans. We want to use some akin to } immersion oil for the interface between rock and flatbed, but want to find } something inexpensive, non-toxic and easy to clean. Any thoughts? } } TIA & genuinely :o) } michael shaffer } } SEM/MLA Research Coordinator } (709) 737-6799 (ofc) } (709) 737-6790 (lab) } (709) 737-6193 (FAX) } {http://www.mun.ca/creait/maf/} } {http://www.esd.mun.ca/epma/} } } Inco Innovation Centre } c/o Memorial University } 230 Elizabeth Avenue } P.O. Box 4200 } St. John's, NL A1C 5S7 } } } } } ==============================Original Headers============================== } 7, 19 -- From Michael-at-Shaffer.net Thu May 25 05:52:02 2006 } 7, 19 -- Received: from ws6-4.us4.outblaze.com (ws6-4.us4.outblaze.com [205.158.62.107]) } 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k4PAq1qK008574 } 7, 19 -- for {Microscopy-at-microscopy.com} ; Thu, 25 May 2006 05:52:01 -0500 } 7, 19 -- Received: (qmail 17426 invoked from network); 25 May 2006 10:52:00 -0000 } 7, 19 -- Received: from unknown (HELO roamingwolf) (Michael-at-Shaffer.net-at-134.153.130.141) } 7, 19 -- by ws6-4.us4.outblaze.com with SMTP; 25 May 2006 10:52:00 -0000 } 7, 19 -- From: "michael shaffer" {michael-at-Shaffer.net} } 7, 19 -- To: "MSA Microscopy list" {Microscopy-at-microscopy.com} } 7, 19 -- Subject: imaging rock core with flatbed scanner } 7, 19 -- Date: Thu, 25 May 2006 08:21:59 -0230 } 7, 19 -- Message-ID: {000f01c67fe9$40c556e0$8d829986-at-roamingwolf} } 7, 19 -- MIME-Version: 1.0 } 7, 19 -- Content-Type: text/plain; } 7, 19 -- charset="US-ASCII" } 7, 19 -- Content-Transfer-Encoding: 7bit } 7, 19 -- X-Mailer: Microsoft Office Outlook 11 } 7, 19 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } 7, 19 -- Thread-Index: AcZ/6T9zCvUQjGd1R8+CGbe9ZHz2pA== } ==============================End of - Headers============================== } }
Greetings listers. We have an image analysis problem, and I could use your help. We are frequently asked to characterize the cell size and structure of various polymer foams for our customers. To date we have relied upon an ASTM method that involves counting the number of cell intersections with a reference line drawn on the image to compute average cell size. The method assumes a degree of homogeneity and cell density that is often an imperfect representation of our customer's samples, and in any event it would be far more desirable to select and measure the cells directly with our ImageProPlus software. There's the rub. We're having the devil of a time getting good segmentation of the open cells against the background matrix. We've tried a number of filters and operations, but thus far without success. I have posted two examples through the link listed below (I disabled it to get through the spam filter). To download, just right click on the file name and "save target as".
**http://www.impactanalytical.com/data/**
If any of you experts out there can set us on the right track, or alternatively explain why we're wasting our time, I would GREATLY appreciate it. My head hurts and the walls are in need of repair!
Incidentally, I have tried varying kV, and collecting in backscatter as well as backscatter plus/minus for topography. Thus far these efforts have not solved the fundamental problem that contrast along the rim of a given cell shifts from light to dark, confounding segmentation. Thanks for your consideration - there's a windy city beer in the bargain if my travel request goes through! Yours, Matt Matthew Stephenson Impact Analytical/MMI 1910 W. Saint Andrews Rd. Midland, MI 48640 (989)832-5555 X506
==============================Original Headers============================== 1, 14 -- From stephenson-at-impactanalytical.com Mon Jun 12 07:19:49 2006 1, 14 -- Received: from mitconexch.mitcon.org (mitconexch.mitcon.org [192.251.56.2]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CCJnEA003629 1, 14 -- for {Microscopy-at-Microscopy.Com} ; Mon, 12 Jun 2006 07:19:49 -0500 1, 14 -- Received: by mitconexch.mitcon.org with Internet Mail Service (5.5.2656.59) 1, 14 -- id {M30M2R2Q} ; Mon, 12 Jun 2006 08:19:47 -0400 1, 14 -- Message-ID: {089C7709BE9235448E3622C6F38D828F06C69196-at-mitconexch.mitcon.org} 1, 14 -- From: "Stephenson, Matthew" {stephenson-at-impactanalytical.com} 1, 14 -- To: "'Microscopy-at-Microscopy.Com'" {Microscopy-at-Microscopy.Com} 1, 14 -- Subject: Image Analysis problem 1, 14 -- Date: Mon, 12 Jun 2006 08:19:46 -0400 1, 14 -- MIME-Version: 1.0 1, 14 -- X-Mailer: Internet Mail Service (5.5.2656.59) 1, 14 -- Content-Type: text/plain ==============================End of - Headers==============================
Your images are nearly identical to one that I use as a lab experiment in the image analysis courses I teach.
If you examine the image visually, how do you "see" the edges of the pores? There is always some local brightness change associated with the edge. A good way to highlight these edges is to duplicate the image, and perform a morphological erosion to replace each pixel with its darkest neighbor. Then subtract that image from the original, which will leave the locallly lightest pixels. These highlight the edges. Of course, you will want to do a bit of cleanup on the images first (a median filter or one of its derivatives is probably better than a smoothing convolution).
Your major problem is not ** measuring ** the circular features in your image, it is relating those measurements to the pores. This is not an imaging problem but a stereological one. The sections through the pores are NOT the pore sizes. There is a lot of literature on the relationships between the sizes of 2D sections through features and the sizes of the 3D structures.
John Russ
-----Original Message----- X-from: stephenson-at-impactanalytical.com To: DrJohnRuss-at-AOL.com Sent: Mon, 12 Jun 2006 07:20:35 -0500
Greetings listers. We have an image analysis problem, and I could use your help. We are frequently asked to characterize the cell size and structure of various polymer foams for our customers. To date we have relied upon an ASTM method that involves counting the number of cell intersections with a reference line drawn on the image to compute average cell size. The method assumes a degree of homogeneity and cell density that is often an imperfect representation of our customer's samples, and in any event it would be far more desirable to select and measure the cells directly with our ImageProPlus software. There's the rub. We're having the devil of a time getting good segmentation of the open cells against the background matrix. We've tried a number of filters and operations, but thus far without success. I have posted two examples through the link listed below (I disabled it to get through the spam filter). To download, just right click on the file name and "save target as".
**http://www.impactanalytical.com/data/**
If any of you experts out there can set us on the right track, or alternatively explain why we're wasting our time, I would GREATLY appreciate it. My head hurts and the walls are in need of repair!
Incidentally, I have tried varying kV, and collecting in backscatter as well as backscatter plus/minus for topography. Thus far these efforts have not solved the fundamental problem that contrast along the rim of a given cell shifts from light to dark, confounding segmentation. Thanks for your consideration - there's a windy city beer in the bargain if my travel request goes through! Yours, Matt Matthew Stephenson Impact Analytical/MMI 1910 W. Saint Andrews Rd. Midland, MI 48640 (989)832-5555 X506
==============================Original Headers============================== 1, 14 -- From stephenson-at-impactanalytical.com Mon Jun 12 07:19:49 2006 1, 14 -- Received: from mitconexch.mitcon.org (mitconexch.mitcon.org [192.251.56.2]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CCJnEA003629 1, 14 -- for {Microscopy-at-Microscopy.Com} ; Mon, 12 Jun 2006 07:19:49 -0500 1, 14 -- Received: by mitconexch.mitcon.org with Internet Mail Service (5.5.2656.59) 1, 14 -- id {M30M2R2Q} ; Mon, 12 Jun 2006 08:19:47 -0400 1, 14 -- Message-ID: {089C7709BE9235448E3622C6F38D828F06C69196-at-mitconexch.mitcon.org} 1, 14 -- From: "Stephenson, Matthew" {stephenson-at-impactanalytical.com} 1, 14 -- To: "'Microscopy-at-Microscopy.Com'" {Microscopy-at-Microscopy.Com} 1, 14 -- Subject: Image Analysis problem 1, 14 -- Date: Mon, 12 Jun 2006 08:19:46 -0400 1, 14 -- MIME-Version: 1.0 1, 14 -- X-Mailer: Internet Mail Service (5.5.2656.59) 1, 14 -- Content-Type: text/plain ==============================End of - Headers==============================
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==============================Original Headers============================== 17, 25 -- From DrJohnRuss-at-aol.com Mon Jun 12 08:09:54 2006 17, 25 -- Received: from imo-m20.mx.aol.com (imo-m20.mx.aol.com [64.12.137.1]) 17, 25 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CD9r0E014603 17, 25 -- for {microscopy-at-sparc5.microscopy.com} ; Mon, 12 Jun 2006 08:09:53 -0500 17, 25 -- Received: from DrJohnRuss-at-aol.com 17, 25 -- by imo-m20.mx.aol.com (mail_out_v38_r7.5.) id v.4c9.f9cb4e (15886); 17, 25 -- Mon, 12 Jun 2006 09:09:47 -0400 (EDT) 17, 25 -- Received: from MBLK-M40 (mblk-m40.mblk.aol.com [64.12.136.84]) by air-id08.mx.aol.com (v109.13) with ESMTP id MAILINID81-3e0e448d679b241; Mon, 12 Jun 2006 09:09:47 -0400 17, 25 -- Content-Transfer-Encoding: 7bit 17, 25 -- Date: Mon, 12 Jun 2006 09:09:47 -0400 17, 25 -- Message-Id: {8C85C389D78B718-17B0-4362-at-MBLK-M40.sysops.aol.com} 17, 25 -- From: drjohnruss-at-aol.com 17, 25 -- References: {200606121220.k5CCKZt8004417-at-ns.microscopy.com} 17, 25 -- Cc: stephenson-at-impactanalytical.com 17, 25 -- Received: from 65.190.140.228 by MBLK-M40.sysops.aol.com (64.12.136.84) with HTTP (WebMailUI); Mon, 12 Jun 2006 09:09:47 -0400 17, 25 -- X-MB-Message-Source: WebUI 17, 25 -- X-MB-Message-Type: User 17, 25 -- In-Reply-To: {200606121220.k5CCKZt8004417-at-ns.microscopy.com} 17, 25 -- X-Mailer: AOL WebMail 17789 17, 25 -- Subject: Re: [Microscopy] Image Analysis problem 17, 25 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 17, 25 -- MIME-Version: 1.0 17, 25 -- To: microscopy-at-ns.microscopy.com 17, 25 -- X-AOL-IP: 64.12.136.84 17, 25 -- X-Spam-Flag: NO ==============================End of - Headers==============================
There have been several good suggestions already, so I will try to add a few hints that have worked for me. First of all, do make sure you are not
milling in the same direction as the epoxy line, as that will preferentially mill the epoxy away. Use beam modulation or the equivalent
to restrict the beam to milling perpendicular to the interface. Also, I've noticed the thinner the epoxy line, the better the milling. When gluing your two sample pieces together, make sure the surfaces are absolutely clean, check the epoxy (I always use G-1) for specks of 'stuff', and use the strongest clamp/vise you can. This will protect the epoxy line as well. Finally, when you do have a hard-to-glue surface, sometimes this can be improved by coating it with a thin layer of a material that will adhere to the G-1, such as chromium. You didn't mention dimpling in your sample prep, but this might also help. In my experience dimpling is less preferential than ion milling, and can help you get a thinner sample that requires less milling.
Leslie
Leslie Krupp (Thompson) IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
jds451-at-psu.edu 06/09/2006 09:49 AM Please respond to jds451-at-psu.edu
To Leslie E Krupp/Almaden/IBM-at-IBMUS cc
Subject [Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep
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Hi all,
I have been attempting to prepare cross sectional TEM samples of anodic tantala (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an approximate thickness of 200um. The oxide film is grown anodically on the
Ta using electrochemistry.
In the literature, I have seen cross sectional TEM images of anodic tantala grown on sputter deposited tantalum; an aluminum or silicon substrate is typically used. I am curious if anyone has had success in preparing anodic Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)
My main problem seems to be with de-lamination of the cross section sandwich. Here is my attempted procedure:
I first bound two faces of my sample together and placed the sandwich in a
press while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich frequently breaks apart when I begin polishing. To address this problem, I encased my sandwich in a brass tube and fitted the tube with supporting brass shims, all held in place with epoxy. Using this technique, I was able to thin some parts of the specimen to 20-50 um, but the epoxy was no longer present at this point. When I attempted to ion-mill, the sandwiches splintered apart, and any of the oxide that may have been surviving was obliterated in the ion mill.
Any suggestions or advice would be greatly appreciated.
Thank You,
Jennifer Ray Sloppy Materials Science & Engineering Department Ph.D. Candidate Pennsylvania State University
==============================Original Headers============================== 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201]) 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752 9, 17 -- for |--microscopy-at-microscopy.com--|; Fri, 9 Jun 2006 11:45:10 -0500 9, 17 -- Received: (from webmail-at-localhost) 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431; 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT) 9, 17 -- Message-Id: |--200606091645.MAA01431-at-webmail15.cac.psu.edu--| 9, 17 -- To: microscopy-at-microscopy.com 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep 9, 17 -- From: "Jen Sloppy" |--jds451-at-psu.edu--| 9, 17 -- X-Mailer: Penn State WebMail 9, 17 -- X-Sender: jds451 9, 17 -- X-Originating-IP: 128.118.37.54 9, 17 -- MIME-Version: 1.0 9, 17 -- Content-Type: text/plain ==============================End of - Headers==============================
|--br--||--font size=2 face="sans-serif"--|Hi Jennifer-|--/font--| |--br--| |--br--||--font size=2 face="sans-serif"--|There have been several good suggestions already, so I will try to add a few hints that have worked for me. First of all, do make sure you are not milling in the same direction as the epoxy line, as that will preferentially mill the epoxy away. Use beam modulation or the equivalent to restrict the beam to milling perpendicular to the interface. Also, I've noticed the thinner the epoxy line, the better the milling. When gluing your two sample pieces together, make sure the surfaces are absolutely clean, check the epoxy (I always use G-1) for specks of 'stuff', and use the strongest clamp/vise you can. This will protect the epoxy line as well. Finally, when you do have a hard-to-glue surface, sometimes this can be improved by coating it with a thin layer of a material that will adhere to the G-1, such as chromium. You didn't mention dimpling in your sample prep, but this might also help. In my experience dimpling is less preferential than ion milling, and can help you get a thinner sample that requires less milling.|--/font--| |--br--| |--br--||--font size=2 face="sans-serif"--|Leslie|--/font--| |--br--| |--br--||--font size=2 face="sans-serif"--|Leslie Krupp (Thompson)|--br--| IBM Almaden Research|--br--| 650 Harry Road, K19/D1|--br--| San Jose, CA 95120-6099|--br--| (408) 927-3856|--/font--| |--br--| |--br--| |--br--| |--table width=100%--| |--tr valign=top--| |--td width=40%--||--font size=1 face="sans-serif"--||--b--|jds451-at-psu.edu|--/b--| |--/font--| |--p--||--font size=1 face="sans-serif"--|06/09/2006 09:49 AM|--/font--| |--table border--| |--tr valign=top--| |--td bgcolor=white--| |--div align=center--||--font size=1 face="sans-serif"--|Please respond to|--br--| jds451-at-psu.edu|--/font--||--/div--||--/table--| |--br--| |--td width=59%--| |--table width=100%--| |--tr valign=top--| |--td--| |--div align=right--||--font size=1 face="sans-serif"--|To|--/font--||--/div--| |--td--||--font size=1 face="sans-serif"--|Leslie E Krupp/Almaden/IBM-at-IBMUS|--/font--| |--tr valign=top--| |--td--| |--div align=right--||--font size=1 face="sans-serif"--|cc|--/font--||--/div--| |--td--| |--tr valign=top--| |--td--| |--div align=right--||--font size=1 face="sans-serif"--|Subject|--/font--||--/div--| |--td--||--font size=1 face="sans-serif"--|[Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep|--/font--||--/table--| |--br--| |--table--| |--tr valign=top--| |--td--| |--td--||--/table--| |--br--||--/table--| |--br--| |--br--| |--br--||--tt--||--font size=2--||--br--| |--br--| |--br--| ----------------------------------------------------------------------------|--br--| The Microscopy ListServer -- CoSponsor: The Microscopy Society of America|--br--| To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver|--br--| On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html|--br--| ----------------------------------------------------------------------------|--br--| |--br--| Hi all,|--br--| |--br--| I have been attempting to prepare cross sectional TEM samples of anodic tantala|--br--| (Ta2O5) by mechanical polishing. The substrate is tantalum metal with an|--br--| approximate thickness of 200um. The oxide film is grown anodically on the Ta|--br--| using electrochemistry. |--br--| |--br--| In the literature, I have seen cross sectional TEM images of anodic tantala|--br--| grown on sputter deposited tantalum; an aluminum or silicon substrate is|--br--| typically used. I am curious if anyone has had success in preparing anodic|--br--| Ta2O5 samples on a tantalum substrate. (not a sputtered tantalum layer)|--br--| |--br--| My main problem seems to be with de-lamination of the cross section sandwich.|--br--| Here is my attempted procedure: |--br--| |--br--| I first bound two faces of my sample together and placed the sandwich in a press|--br--| while the epoxy cured (I have tried M-Bond and G1 from Gatan), but the sandwich|--br--| frequently breaks apart when I begin polishing. To address this problem, I|--br--| encased my sandwich in a brass tube and fitted the tube with supporting brass|--br--| shims, all held in place with epoxy. Using this technique, I was able to thin|--br--| some parts of the specimen to 20-50 um, but the epoxy was no longer present at|--br--| this point. When I attempted to ion-mill, the sandwiches splintered apart, and|--br--| any of the oxide that may have been surviving was obliterated in the ion mill.|--br--| |--br--| Any suggestions or advice would be greatly appreciated.|--br--| |--br--| Thank You,|--br--| |--br--| Jennifer Ray Sloppy|--br--| Materials Science & Engineering Department|--br--| Ph.D. Candidate|--br--| Pennsylvania State University|--br--| |--br--| |--br--| ==============================Original Headers==============================|--br--| 9, 17 -- From jds451-at-psu.edu Fri Jun 9 11:45:10 2006|--br--| 9, 17 -- Received: from webmail15.cac.psu.edu (webmail15.cac.psu.edu [128.118.1.201])|--br--| 9, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k59GjAoI029752|--br--| 9, 17 -- for <microscopy-at-microscopy.com>; Fri, 9 Jun 2006 11:45:10 -0500|--br--| 9, 17 -- Received: (from webmail-at-localhost)|--br--| 9, 17 -- by webmail15.cac.psu.edu (8.9.3/8.9.0) id MAA01431;|--br--| 9, 17 -- Fri, 9 Jun 2006 12:45:09 -0400 (EDT)|--br--| 9, 17 -- Date: Fri, 9 Jun 2006 12:45:09 -0400 (EDT)|--br--| 9, 17 -- Message-Id: <200606091645.MAA01431-at-webmail15.cac.psu.edu>|--br--| 9, 17 -- To: microscopy-at-microscopy.com|--br--| 9, 17 -- Subject: Tantalum / Ta Oxide Cross Section TEM Sample Prep|--br--| 9, 17 -- From: "Jen Sloppy" <jds451-at-psu.edu>|--br--| 9, 17 -- X-Mailer: Penn State WebMail|--br--| 9, 17 -- X-Sender: jds451|--br--| 9, 17 -- X-Originating-IP: 128.118.37.54|--br--| 9, 17 -- MIME-Version: 1.0|--br--| 9, 17 -- Content-Type: text/plain|--br--| ==============================End of - Headers==============================|--br--| |--/font--||--/tt--| |--br--| --=_alternative 005BDC628825718B_=--
==============================Original Headers============================== 33, 44 -- From lkrupp-at-us.ibm.com Mon Jun 12 11:45:33 2006 33, 44 -- Received: from e2.ny.us.ibm.com (e2.ny.us.ibm.com [32.97.182.142]) 33, 44 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CGjWFJ030320 33, 44 -- for |--Microscopy-at-microscopy.com--|; Mon, 12 Jun 2006 11:45:33 -0500 33, 44 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 33, 44 -- by e2.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CGjWqG004007 33, 44 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 33, 44 -- for |--Microscopy-at-microscopy.com--|; Mon, 12 Jun 2006 12:45:32 -0400 33, 44 -- Received: from d01av04.pok.ibm.com (d01av04.pok.ibm.com [9.56.224.64]) 33, 44 -- by d01relay02.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k5CGjW0x268382 33, 44 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 33, 44 -- for |--Microscopy-at-microscopy.com--|; Mon, 12 Jun 2006 12:45:32 -0400 33, 44 -- Received: from d01av04.pok.ibm.com (loopback [127.0.0.1]) 33, 44 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k5CGjVvI008729 33, 44 -- for |--Microscopy-at-microscopy.com--|; Mon, 12 Jun 2006 12:45:31 -0400 33, 44 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 33, 44 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CGjVZ8008714; 33, 44 -- Mon, 12 Jun 2006 12:45:31 -0400 33, 44 -- In-Reply-To: |--200606091649.k59GnIlh004856-at-ns.microscopy.com--| 33, 44 -- MIME-Version: 1.0 33, 44 -- X-MIMETrack: S/MIME Sign by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF144|February 33, 44 -- 01, 2006) at 06/12/2006 09:43:15 AM, 33, 44 -- Serialize by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF144|February 33, 44 -- 01, 2006) at 06/12/2006 09:43:15 AM, 33, 44 -- Serialize complete at 06/12/2006 09:43:15 AM, 33, 44 -- S/MIME Sign failed at 06/12/2006 09:43:15 AM: The cryptographic key was not 33, 44 -- found, 33, 44 -- S/MIME Sign by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF144|February 33, 44 -- 01, 2006) at 06/12/2006 09:43:22 AM, 33, 44 -- Serialize by Notes Client on Leslie E Krupp/Almaden/IBM(Release 7.0 HF144|February 33, 44 -- 01, 2006) at 06/12/2006 09:43:22 AM, 33, 44 -- Serialize complete at 06/12/2006 09:43:22 AM, 33, 44 -- S/MIME Sign failed at 06/12/2006 09:43:22 AM: The cryptographic key was not 33, 44 -- found, 33, 44 -- Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF123 | April 14, 2006) at 33, 44 -- 06/12/2006 12:45:30, 33, 44 -- Serialize complete at 06/12/2006 12:45:30 33, 44 -- To: jds451-at-psu.edu, Microscopy-at-microscopy.com 33, 44 -- Subject: Re: [Microscopy] Tantalum / Ta Oxide Cross Section TEM Sample Prep 33, 44 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 33, 44 -- Message-ID: |--OF2B7D685D.EA6215AC-ON8525718B.005B2D78-8825718B.005BDC65-at-us.ibm.com--| 33, 44 -- From: Leslie E Krupp |--lkrupp-at-us.ibm.com--| 33, 44 -- Date: Mon, 12 Jun 2006 12:45:27 -0400 33, 44 -- Content-Type: multipart/alternative; boundary="=_alternative 005BDC628825718B_=" ==============================End of - Headers==============================
==============================Original Headers============================== 37, 29 -- From lkrupp-at-us.ibm.com Mon Jun 12 11:52:44 2006 37, 29 -- Received: from e2.ny.us.ibm.com (e2.ny.us.ibm.com [32.97.182.142]) 37, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CGqhxa030442 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 11:52:43 -0500 37, 29 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 37, 29 -- by e2.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CGqhDZ018610 37, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:52:43 -0400 37, 29 -- Received: from d01av04.pok.ibm.com (d01av04.pok.ibm.com [9.56.224.64]) 37, 29 -- by d01relay04.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k5CGqhL1140474 37, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:52:43 -0400 37, 29 -- Received: from d01av04.pok.ibm.com (loopback [127.0.0.1]) 37, 29 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k5CGqgWC001568 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:52:42 -0400 37, 29 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 37, 29 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CGqgYV001558 37, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:52:42 -0400 37, 29 -- To: microscopy-at-microscopy.com 37, 29 -- Subject: Re: Tantalum / Ta Oxide Cross Section TEM Sample Prep 37, 29 -- MIME-Version: 1.0 37, 29 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 37, 29 -- Message-ID: {OF138984EA.85B0B778-ON8525718B.005C97EB-8825718B.005D1109-at-us.ibm.com} 37, 29 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 37, 29 -- Date: Mon, 12 Jun 2006 09:52:41 -0700 37, 29 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF123 | April 14, 2006) at 37, 29 -- 06/12/2006 12:52:42, 37, 29 -- Serialize complete at 06/12/2006 12:52:42 37, 29 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
Many thanks to all the folks who responded to my post. Here is a summary of the suggestions.
At least 4 or 5 people suggested sonicating the powders in a solvent prior to disbursing. After this step, some suggested putting a drop on a filmed grid, or using some type of atomizer to spray the dispersion onto a grid. (Perfume sprayer, air can held in a jar, nebulizer)
A couple people recommended simply mixing the powder with a solvent without sonicating and either letting a drop dry on the grid, or heating it to aid evaporation.
There were two mentions of dry loading the grid, which is dumping some of the powder on the grid and tapping off the excess. This is the method I tried first as it was the easiest, but I have not looked at the grid yet to see if there is enough material to do EELS on.
Thanks again, Leslie
Leslie Krupp (Thompson) IBM Almaden Research 650 Harry Road, K19/D1 San Jose, CA 95120-6099 (408) 927-3856
==============================Original Headers============================== 7, 29 -- From lkrupp-at-us.ibm.com Mon Jun 12 13:11:36 2006 7, 29 -- Received: from e4.ny.us.ibm.com (e4.ny.us.ibm.com [32.97.182.144]) 7, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CIBZl6009945 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 13:11:36 -0500 7, 29 -- Received: from d01relay02.pok.ibm.com (d01relay02.pok.ibm.com [9.56.227.234]) 7, 29 -- by e4.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CIBZMS030830 7, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:11:35 -0400 7, 29 -- Received: from d01av04.pok.ibm.com (d01av04.pok.ibm.com [9.56.224.64]) 7, 29 -- by d01relay02.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k5CIA9EN157808 7, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:10:09 -0400 7, 29 -- Received: from d01av04.pok.ibm.com (loopback [127.0.0.1]) 7, 29 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k5CIA9B0006658 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:10:09 -0400 7, 29 -- Received: from d01ml604.pok.ibm.com (d01ml604.pok.ibm.com [9.56.227.90]) 7, 29 -- by d01av04.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5CIA8Mr006651 7, 29 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:10:08 -0400 7, 29 -- To: microscopy-at-microscopy.com 7, 29 -- Subject: Powder sample prep summary 7, 29 -- MIME-Version: 1.0 7, 29 -- X-Mailer: Lotus Notes Release 7.0 HF144 February 01, 2006 7, 29 -- Message-ID: {OF7E95A364.DAB1C69A-ON8525718B.005CF6C7-8825718B.00642817-at-us.ibm.com} 7, 29 -- From: Leslie E Krupp {lkrupp-at-us.ibm.com} 7, 29 -- Date: Mon, 12 Jun 2006 11:10:07 -0700 7, 29 -- X-MIMETrack: Serialize by Router on D01ML604/01/M/IBM(Release 7.0.1HF123 | April 14, 2006) at 7, 29 -- 06/12/2006 14:10:08, 7, 29 -- Serialize complete at 06/12/2006 14:10:08 7, 29 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
Great suggestions, John, I will give them a try. And point well taken regarding the indirect relation of the 2D feature to the 3D pore size. The ASTM method we use (#3576) makes that dimensional correction, but only by assuming close-packed, spherical features. That said, our customers have real use for comparative data from their less ideal samples. If we can select their 2D open cells, they can compare one region to another as a function of molding conditions, one sample to another as a function of processing or R&D variations, or see how they stack up with the competition. But if I can get your suggestions to work, we can offer them stereological analysis as well. Just have to see if they're willing to pay...Thanks again! Yours, Matt
-----Original Message----- X-from: drjohnruss-at-aol.com [mailto:drjohnruss-at-aol.com] Sent: Monday, June 12, 2006 9:10 AM To: microscopy-at-sparc5.microscopy.com Cc: stephenson-at-impactanalytical.com
Greetings listers. We have an image analysis problem, and I could use your help. We are frequently asked to characterize the cell size and structure of various polymer foams for our customers. To date we have relied upon an ASTM method that involves counting the number of cell intersections with a reference line drawn on the image to compute average cell size. The method assumes a degree of homogeneity and cell density that is often an imperfect representation of our customer's samples, and in any event it would be far more desirable to select and measure the cells directly with our ImageProPlus software. There's the rub. We're having the devil of a time getting good segmentation of the open cells against the background matrix. We've tried a number of filters and operations, but thus far without success. I have posted two examples through the link listed below (I disabled it to get through the spam filter). To download, just right click on the file name and "save target as".
**http://www.impactanalytical.com/data/**
If any of you experts out there can set us on the right track, or alternatively explain why we're wasting our time, I would GREATLY appreciate it. My head hurts and the walls are in need of repair!
Incidentally, I have tried varying kV, and collecting in backscatter as well as backscatter plus/minus for topography. Thus far these efforts have not solved the fundamental problem that contrast along the rim of a given cell shifts from light to dark, confounding segmentation. Thanks for your consideration - there's a windy city beer in the bargain if my travel request goes through! Yours, Matt Matthew Stephenson Impact Analytical/MMI 1910 W. Saint Andrews Rd. Midland, MI 48640 (989)832-5555 X506
==============================Original Headers============================== 1, 14 -- From stephenson-at-impactanalytical.com Mon Jun 12 07:19:49 2006 1, 14 -- Received: from mitconexch.mitcon.org (mitconexch.mitcon.org [192.251.56.2]) 1, 14 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CCJnEA003629 1, 14 -- for {Microscopy-at-Microscopy.Com} ; Mon, 12 Jun 2006 07:19:49 -0500 1, 14 -- Received: by mitconexch.mitcon.org with Internet Mail Service (5.5.2656.59) 1, 14 -- id {M30M2R2Q} ; Mon, 12 Jun 2006 08:19:47 -0400 1, 14 -- Message-ID: {089C7709BE9235448E3622C6F38D828F06C69196-at-mitconexch.mitcon.org} 1, 14 -- From: "Stephenson, Matthew" {stephenson-at-impactanalytical.com} 1, 14 -- To: "'Microscopy-at-Microscopy.Com'" {Microscopy-at-Microscopy.Com} 1, 14 --
Hi All,
I have a user who has presented a very tough problem.
They are looking at a polymer tube (ID ~ .1 - 1 micron) which is filled with a liquid with CNT suspended in it. The tube is the result of drawing from a larger tube and they are hoping to have aligned CNT. They would like to verify this in some way.
I have access to a FESEM and a conventional TEM. The SEM is not environmental. I could not come up with a way to image these. The SEM will not be able to resolve the CNT in any case I think. The TEM could but the sample would have to be thinned to the point that it is likely the inner core of the tube is penetrated. I don't think liquid in the TEM is viable.
I have suggested XRD as a possibility, but the CNT signal on top of the tube and liquid signal may be problematic.
Any thoughts?
Thanks,
Tom
------------------------------------------------------------------------ --------------------------------------------- Thomas M Murray email: murraytm-at-u.washington.edu Electron Microscopy Center Manager Phone: (206)543-2836 Materials Science & Engineering Fax: (206)543-3100 Box 352120 302 Roberts Hall Cell: (425)345-0083 University of Washington Seattle, WA 98195
==============================Original Headers============================== 10, 23 -- From murraytm-at-u.washington.edu Mon Jun 12 14:42:08 2006 10, 23 -- Received: from mxout7.cac.washington.edu (mxout7.cac.washington.edu [140.142.32.178]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CJg7v8000335 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 14:42:08 -0500 10, 23 -- Received: from smtp.washington.edu (smtp.washington.edu [140.142.33.9]) 10, 23 -- by mxout7.cac.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k5CJg7wC030917 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:42:07 -0700 10, 23 -- X-Auth-Received: from [128.95.118.89] (tstoebe-mse.ad.engr.washington.edu [128.95.118.89]) 10, 23 -- (authenticated authid=murraytm) 10, 23 -- by smtp.washington.edu (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k5CJg4aU013369 10, 23 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 12:42:07 -0700 10, 23 -- Mime-Version: 1.0 (Apple Message framework v749.3) 10, 23 -- X-Priority: 3 10, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 10, 23 -- Message-Id: {427C2B76-E9DC-4F69-9B49-0256CEBCB34D-at-u.washington.edu} 10, 23 -- Content-Transfer-Encoding: 7bit 10, 23 -- From: Tom Murray {murraytm-at-u.washington.edu} 10, 23 -- Subject: Imaging Carbon Nanotubes 10, 23 -- Date: Mon, 12 Jun 2006 12:42:03 -0700 10, 23 -- To: "Microscopy ListServer (E-mail)" {Microscopy-at-microscopy.com} 10, 23 -- X-Mailer: Apple Mail (2.749.3) 10, 23 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_MEDIA_BODY 0, __CP_NOT_1 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __HAS_X_PRIORITY 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0, __STOCK_CRUFT 0' ==============================End of - Headers==============================
Any suggestions on the best way to image protein crystals? They are 20-100 um in length and very stable in high molarity salt solutions or PEG solutions, but dissolve readily in less dilute aqueous solutions. We would like to avoid chemical fixation.
Joe Neilly
==============================Original Headers============================== 2, 18 -- From joe.p.neilly-at-abbott.com Mon Jun 12 15:33:39 2006 2, 18 -- Received: from abtmx3.abbott.com (abtmx3.abbott.com [130.36.44.93]) 2, 18 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CKXdov011623 2, 18 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 15:33:39 -0500 2, 18 -- Received: from abtapn561.northamerica.intra.abbott.com (abtapn561.northamerica.intra.abbott.com [10.236.188.103]) 2, 18 -- by abtmx3.abbott.com (Switch-3.1.8/Switch-3.1.7) with ESMTP id k5CKXcRN008161 2, 18 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 15:33:38 -0500 (CDT) 2, 18 -- To: microscopy-at-microscopy.com 2, 18 -- Subject: Imaging Protein Crystals 2, 18 -- MIME-Version: 1.0 2, 18 -- X-Mailer: Lotus Notes Release 6.5.4 HF761 November 21, 2005 2, 18 -- Message-ID: {OF00F34D2B.F71960B2-ON8625718B.00707FED-8625718B.0070F0E1-at-abbott.com} 2, 18 -- From: Joseph P Neilly {joe.p.neilly-at-abbott.com} 2, 18 -- Date: Mon, 12 Jun 2006 15:33:01 -0500 2, 18 -- X-MIMETrack: Serialize by Router on ABTAPN561/ESVR/ABBOTT(Release 6.5.4FP2|September 12, 2005) at 2, 18 -- 06/12/2006 15:33:33, 2, 18 -- Serialize complete at 06/12/2006 15:33:33 2, 18 -- Content-Type: text/plain; charset="US-ASCII" ==============================End of - Headers==============================
Greetings, If the goal is not to image the tubes per se but to tell whether they have a net alignment, polarized light microscopy could work. I think sensitivity is ok (With a good instrument, you can detect birefringent retardation of 1 nm and with a really good instrument you can get down to almost 0.1 nm). I expect the problem will come from the polymer tube. Its material could be birefringent and if the inner diameter is really 0.1 micron (rather than 1 micron) then that will probably be too small. But if your i.d.s are closer to 1 micron and the polymer tube itself behaves in the light, it could work...
Good luck! Tobias } } } Hi All, } } I have a user who has presented a very tough problem. } } They are looking at a polymer tube (ID ~ .1 - 1 micron) which is } filled with a liquid with CNT suspended in it. The tube is the } result of drawing from a larger tube and they are hoping to have } aligned CNT. They would like to verify this in some way. } } I have access to a FESEM and a conventional TEM. The SEM is not } environmental. I could not come up with a way to image these. The } SEM will not be able to resolve the CNT in any case I think. The TEM } could but the sample would have to be thinned to the point that it is } likely the inner core of the tube is penetrated. I don't think } liquid in the TEM is viable. } } I have suggested XRD as a possibility, but the CNT signal on top of } the tube and liquid signal may be problematic. } } Any thoughts? } } Thanks, } } Tom } } ------------------------------------------------------------------------ } --------------------------------------------- } Thomas M Murray email: } murraytm-at-u.washington.edu } Electron Microscopy Center Manager Phone: (206)543-2836 } Materials Science & Engineering Fax: (206)543-3100 } Box 352120 302 Roberts Hall Cell: (425)345-0083 } University of Washington } Seattle, WA 98195 } } } ==============================Original Headers============================== } 10, 23 -- From murraytm-at-u.washington.edu Mon Jun 12 14:42:08 2006 } 10, 23 -- Received: from mxout7.cac.washington.edu } (mxout7.cac.washington.edu [140.142.32.178]) } 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k5CJg7v8000335 } 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 } 14:42:08 -0500 } 10, 23 -- Received: from smtp.washington.edu (smtp.washington.edu } [140.142.33.9]) } 10, 23 -- by mxout7.cac.washington.edu } (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k5CJg7wC030917 } 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 } 12:42:07 -0700 } 10, 23 -- X-Auth-Received: from [128.95.118.89] } (tstoebe-mse.ad.engr.washington.edu [128.95.118.89]) } 10, 23 -- (authenticated authid=murraytm) } 10, 23 -- by smtp.washington.edu } (8.13.6+UW06.03/8.13.6+UW06.03) with ESMTP id k5CJg4aU013369 } 10, 23 -- (version=TLSv1/SSLv3 cipher=RC4-SHA bits=128 verify=NOT) } 10, 23 -- for {Microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 } 12:42:07 -0700 } 10, 23 -- Mime-Version: 1.0 (Apple Message framework v749.3) } 10, 23 -- X-Priority: 3 } 10, 23 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed } 10, 23 -- Message-Id: {427C2B76-E9DC-4F69-9B49-0256CEBCB34D-at-u.washington.edu} } 10, 23 -- Content-Transfer-Encoding: 7bit } 10, 23 -- From: Tom Murray {murraytm-at-u.washington.edu} } 10, 23 -- Subject: Imaging Carbon Nanotubes } 10, 23 -- Date: Mon, 12 Jun 2006 12:42:03 -0700 } 10, 23 -- To: "Microscopy ListServer (E-mail)" {Microscopy-at-microscopy.com} } 10, 23 -- X-Mailer: Apple Mail (2.749.3) } 10, 23 -- X-Uwash-Spam: Gauge=IIIIIII, Probability=7%, } Report='__C230066_P5 0, __CP_MEDIA_BODY 0, __CP_NOT_1 0, __CT 0, } __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, } __HAS_X_PRIORITY 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, } __SANE_MSGID 0, __STOCK_CRUFT 0' } ==============================End of - Headers==============================
On Jun 12, 2006, at 1:33 PM, joe.p.neilly-at-abbott.com wrote:
} Any suggestions on the best way to image protein crystals? They are } 20-100 um in length and very stable in high molarity salt solutions or } PEG } solutions, but dissolve readily in less dilute aqueous solutions. We } would like to avoid chemical fixation. } Dear Joe, CryoTEM is a good method if the crystals are thin enough--10s of nm. They would appear lighter against the darker buffer if the salt concentration is high enough, but they might have very little contrast in a PEG buffer. If the crystals are well-ordered, you should see strong diffraction spots and strong spots in the FFT of an image. Plunge freezing would be good for thin crystals. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Mon Jun 12 16:25:51 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CLPoGs032600 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 12 Jun 2006 16:25:51 -0500 4, 22 -- Received: from water-dog.caltech.edu (water-dog [192.168.1.26]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 438BB109B07 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 12 Jun 2006 14:25:50 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by fire-ox.its.caltech.edu (Postfix) with ESMTP id E83C33546E 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Mon, 12 Jun 2006 14:25:43 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200606122033.k5CKXmWv011796-at-ns.microscopy.com} 4, 22 -- References: {200606122033.k5CKXmWv011796-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {7af473644f8ee4dcc48438e217b44feb-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] Imaging Protein Crystals 4, 22 -- Date: Mon, 12 Jun 2006 14:37:12 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on water-dog by amavisd-2.3.3 ==============================End of - Headers==============================
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==============================Original Headers============================== 9, 16 -- From zaluzec-at-aaem.amc.anl.gov Mon Jun 12 16:51:05 2006 9, 16 -- Received: from aaem.amc.anl.gov (aaem.amc.anl.gov [146.139.72.3]) 9, 16 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5CLp3Ho010662 9, 16 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 16:51:04 -0500 9, 16 -- Received: from [146.139.72.105] (aem105.amc.anl.gov [146.139.72.105]) 9, 16 -- by aaem.amc.anl.gov (8.12.11.20060308/8.12.10) with ESMTP id k5CLouFN027186 9, 16 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 16:50:57 -0500 9, 16 -- Mime-Version: 1.0 9, 16 -- Message-Id: {p06110404c0b391c03333-at-[146.139.72.105]} 9, 16 -- In-Reply-To: {200606122033.k5CKXdgd011630-at-ns.microscopy.com} 9, 16 -- References: {200606122033.k5CKXdgd011630-at-ns.microscopy.com} 9, 16 -- Date: Mon, 12 Jun 2006 16:50:55 -0500 9, 16 -- To: microscopy-at-microscopy.com 9, 16 -- From: "Nestor J. Zaluzec" {zaluzec-at-aaem.amc.anl.gov} 9, 16 -- Subject: Re: [Microscopy] Imaging Protein Crystals 9, 16 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (mhunt-at-flinnsci.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Friday, May 26, 2006 at 09:59:18 ---------------------------------------------------------------------------
Email: mhunt-at-flinnsci.com Name: Maureen Hunt
Organization: Flinn
Education: 9-12th Grade High School
Location: Batavia, Illinois, USA
Question: I am a former materials microscopist and teacher who is trying to find a more exciting way to teach mitosis and meiosis. Currently, everyone uses prepared slides of whitefish blastula to teach animal mitosis and either prepared slides or unprepared slides of allium root tips to teach plant mitosis. Is there a protist or other "animal-like" eukaryote that can be given to the students, squashed and stained to show animal mitosis? I have googled and come up short. Any suggestions? Thank-you for your time.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sullivanmassi-at-comcast.net) from http://microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 12, 2006 at 09:12:29 ---------------------------------------------------------------------------
Question: Do you have any recommendations on purchasing microscopes for high school biology? Is there a particular manufacturer that you would go with in terms of value, reliability, etc? Are there new technologies that would fall within a typical high school budget to consider?
Since MItosis and Meiosis are both dynamic processes, I would recommend that you look for movies of the processes.I did a Google search on "mitosis video", and came up with about 300,000 hits. The first few looked quite impressive.
I would also recommend that you teach the function rather than the structure. That is, mitosis functions in normal organismal growth, while meiosis has a significant role only in reproduction.
I can't resist one story. After a lecture that I gave on mitosis in a non-majors Biology course, a student came up to me and said "You have finally cleared up a major mystery for me. I've always wondered how it is that the skin completely covers a little baby, and that it still completely covers the adult that the baby becomes. Now I understand"
Date sent: Mon, 12 Jun 2006 22:04:50 -0500 To: jbs-at-temple.edu X-from: mhunt-at-flinnsci.com Send reply to: mhunt-at-flinnsci.com
This is an extremely broad and unbounded question. How much money are you willing to spend and what analysis methods do you want (LM, DF, DIC, PH, POL, etc.)?
I would recommend querying the usenet group about your question, as it comes up frequently.
sci.techniques.microscopy
Many Web apps have Usenet avenues. There is a large number of informed and experienced folks who deal with LMs on a daily basis. Many are quite helpful for non-professional applications. This is good since the opposite can get out of hand for some folks (i.e., too pricey).
gary g.
At 08:07 PM 6/12/2006, you wrote:
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==============================Original Headers============================== 12, 21 -- From gary-at-gaugler.com Mon Jun 12 23:09:20 2006 12, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5D49KoL020603 12, 21 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 23:09:20 -0500 12, 21 -- Received: (qmail 22096 invoked from network); 12 Jun 2006 21:09:19 -0700 12, 21 -- Received: by simscan 1.1.0 ppid: 22093, pid: 22094, t: 0.1669s 12, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 21 -- by qsmtp3 with SMTP; 12 Jun 2006 21:09:19 -0700 12, 21 -- Message-Id: {7.0.1.0.2.20060612210013.024ea218-at-gaugler.com} 12, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 21 -- Date: Mon, 12 Jun 2006 21:09:21 -0700 12, 21 -- To: sullivanmassi-at-comcast.net 12, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 21 -- Subject: Re: [Microscopy] AskAMicroscopist: purchasing microscopes for 12, 21 -- high school biology 12, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 21 -- In-Reply-To: {200606130307.k5D37AWv001585-at-ns.microscopy.com} 12, 21 -- References: {200606130307.k5D37AWv001585-at-ns.microscopy.com} 12, 21 -- Mime-Version: 1.0 12, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
This Question was submitted to Ask-A-Microscopist by (Goinoutonalamb-at-yahoo.com) from http://www.microscopy.org/Ask-A-Microscopist/Ask-A-Microscopist.html on Monday, June 12, 2006 at 23:04:35 Remember to consider the Grade/Age of the student when considering the Question --------------------------------------------------------------------------- Please reply to both Goinoutonalamb-at-yahoo.com as well as to the Microscopy Listserver ---------------------------------------------------------------------------
Email: Goinoutonalamb-at-yahoo.com Name: Michael Lamb
Organization: Nassau Community College
Education: Undergraduate College
Location: Upton, New York
Title: Tomographic Reconstruction
Question: I am collaborating with a group on a 3D visualization project through Brookhaven National Labs and I have a question pertaining to 3D reconstruction by means of electron tomography. From the literature that I have read, I've gathered that it's recommended that a 200 kv beam or higher is used when working with sections of 120 nm or thicker. Unfortunately, the philips EM-300 that we are working with is only capable of producing a 100 kv beam. Because of the time constraints of our project (10 weeks) it will be very difficult to determine what section thicknesses/maximum tilt angles will work with 100kv with trial and error. Is there a formula, or a guideline of some sort to follow to determine how much mass a 100kv beam can pass with minimal loss of resolution? We are planning on doing a tilt series from -60 to +60 degrees, in 2 degree increments, at 120 nm sections (hopefully). If anyone could lend me some insight as to whether or not this is feasible, and how far we can go, I would greatly appreciate it!
I'm sure there are several articles (also in books, like the one from Joachim Frank on electron tomography) in which the problem of specimen thickness vs. resolution has been discussed.
see e.g. the following article:
Biophys J, February 1998, p. 1031-1042, Vol. 74, No. 2
Electron Tomography of Ice-Embedded Prokaryotic Cells Rudo Grimm,* Hapreet Singh,* Reinhard Rachel,# Dieter Typke,* Wolfram Zillig,* and Wolfgang Baumeister*
best regards, Reinhard Rachel --------------------- PD Dr.Reinhard Rachel Universität Regensburg Zentrum für EM der NWF III am Lehrstuhl fuer Anatomie D-93053 Regensburg - Germany fon +49 941 943 1666, 1720 fax +49 941 943 2868
==============================Original Headers============================== 7, 27 -- From reinhard.rachel-at-biologie.uni-regensburg.de Tue Jun 13 01:49:39 2006 7, 27 -- Received: from rrzmta2.rz.uni-regensburg.de (rrzmta2.rz.uni-regensburg.de [132.199.1.17]) 7, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5D6ncdB017577 7, 27 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 01:49:39 -0500 7, 27 -- Received: from rrzmta2.rz.uni-regensburg.de (localhost [127.0.0.1]) 7, 27 -- by localhost (Postfix) with SMTP id 707C3432F3; 7, 27 -- Tue, 13 Jun 2006 08:49:40 +0200 (CEST) 7, 27 -- Received: from listserv.uni-regensburg.de (titan5.rz.uni-regensburg.de [132.199.4.105]) 7, 27 -- by rrzmta2.rz.uni-regensburg.de (Postfix) with ESMTP id 59096434F0; 7, 27 -- Tue, 13 Jun 2006 08:49:40 +0200 (CEST) 7, 27 -- Received: from TITAN5/SpoolDir by listserv.uni-regensburg.de (Mercury 1.48); 7, 27 -- 13 Jun 06 08:49:38 +0200 7, 27 -- Received: from SpoolDir by TITAN5 (Mercury 1.48); 13 Jun 06 08:49:23 +0200 7, 27 -- From: "Reinhard Rachel" {reinhard.rachel-at-biologie.uni-regensburg.de} 7, 27 -- Organization: Universitaet Regensburg 7, 27 -- To: microscopy-at-microscopy.com 7, 27 -- Date: Tue, 13 Jun 2006 08:49:18 +0200 7, 27 -- MIME-Version: 1.0 7, 27 -- Subject: Tomographic Reconstruction: section thickness 7, 27 -- Cc: Goinoutonalamb-at-yahoo.com 7, 27 -- Priority: normal 7, 27 -- X-mailer: Pegasus Mail for Windows (4.31) 7, 27 -- Content-type: text/plain; charset=ISO-8859-1 7, 27 -- Content-description: Mail message body 7, 27 -- Message-Id: {20060613064940.59096434F0-at-rrzmta2.rz.uni-regensburg.de} 7, 27 -- Content-Transfer-Encoding: 8bit 7, 27 -- X-MIME-Autoconverted: from Quoted-printable to 8bit by ns.microscopy.com id k5D6ncdB017577 ==============================End of - Headers==============================
Probably the most interesting way to teach mitosis is to synchronize cells in culture. It is probably not the easiest though ;-). It is probable that unsynchronized but very actively growing cells will show enough mitotic events to cover the whole cycle. Most of the time they are easy to recognize by normal phase constrast, although fluorescent staining like DAPI are very fast to perform and very impressive to observe. I must admit that after working now for more than 10 years with cells in culture, seeing them split in late telophase is still an exciting event. I witness events which are actually occuring in my own body at that very moment!!!
I agree that you should insist on the functional part though. It is not easy to comprehend these phenomenon in themself, but understand the difference between mitosis and meiosis is really a hard task.
Good luck
Stephane
--- mhunt-at-flinnsci.com wrote:
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==============================Original Headers============================== 10, 20 -- From nizets2-at-yahoo.com Tue Jun 13 03:02:15 2006 10, 20 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5D82EIE028952 10, 20 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 03:02:15 -0500 10, 20 -- Received: (qmail 61561 invoked by uid 60001); 13 Jun 2006 08:02:14 -0000 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 10, 20 -- s=s1024; d=yahoo.com; 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 10, 20 -- b=tOIkUDC8s+gOtnIjmKkJvWbDTGAQj+LFZnsv1ozdZzAzDVLXwfVMD/L2Wg73iyegOLD44ZKSqYVGjNScvzxwgYuXu9AWm8tLly+hjFfmeL0wkxd9mT+mpTwPjeXPGCz7M934lDezDdmiDR+9C3R1aYtmYl6GWYsoE0iyVEXRny4= ; 10, 20 -- Message-ID: {20060613080214.61559.qmail-at-web37413.mail.mud.yahoo.com} 10, 20 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Tue, 13 Jun 2006 01:02:14 PDT 10, 20 -- Date: Tue, 13 Jun 2006 01:02:14 -0700 (PDT) 10, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} 10, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: trying to find a more exciting way to teach 10, 20 -- To: mhunt-at-flinnsci.com 10, 20 -- Cc: microscopy-at-microscopy.com 10, 20 -- In-Reply-To: {200606130311.k5D3B2ck012854-at-ns.microscopy.com} 10, 20 -- MIME-Version: 1.0 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 10, 20 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.com/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both iztok.dogsa-at-ijs.si as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: iztok.dogsa-at-ijs.si Name: iztok dogsa
Organization: IJS
Title-Subject: [Filtered] confocal spinning disk microscope
Question: Has anyone experince with confocal spinning disk microscopy?
Which system would you suggest?
We'd like to achieve really high frame rates, but don't want to buy the laser. The arc lamp is Ok for us.
What about BD CARV II imaging system? Is it better than olympus DSU?
We get asked this fairly regularly by schools and parents (so we have stock replies). Often professional microscopists aren't the best source of information for cheap microscope's as all our optical systems are always over £10,000, and most lab mainstays like our three confocal microscopes come in at nearer £200,000 each (and users still complain about image quality).
Cheap microscopes under £100 are always a disappointment and toy (often called student) microscopes can be very poor. You can easily buy second-hand via ebay, but again there are risks that the optics will be damaged or simply very dirty and difficult to clean (look for old branded 'laboratory' microscopes e.g. Bausch & Lomb) and besides you can't really fill a classroom quickly that way, just get a good one for demos. Local labs may have a good old microscope they no longer need and they may give it to schools (or sell them cheaply) if you acknowledge them - do ask parents and write begging letters if you any spare time (but don't expect anything - look for old establishments that will have years of stuff accumulated). Also try any local microscope enthusiast clubs (they may even be persuaded to come in and talk/demo to pupils) and visit other schools for info and a try-out.
There are suppliers geared up to providing microscopes for schools, so ask around at other school's science departments, but expect to pay nearer £500 each for a quality setup (although with those like bottom end Meade [www.meade.com] at around £100 you can see something at low mag (~20x objective i.e. around 180x mag) with a quality stained section. For pond life etc. a stereoscopic 'dissecting' microscope (40x to 120x mag) is ideal, and of course you can get a really long way with a good magnifying glass (not the really small hi-mag cheap lens ones, try before you buy) - I have a few excellent ones at home for £1 and a good low mag Osram one that includes an illuminating halogen bulb at £8.
Generally prepared slides can rapidly get a bit boring for under 14s, but living or unusual things always attract an audience. Also try your flatbed film scanner (not LiDe and from around £60 upwards) that will be good for looking at soft static things: leaves, fruit, nuts, household objects (scan at max resolution and try reflected and 'film' mode). In the UK there are sites like http://www.wedgwood-group.com/microscopes_digital.htm that cater for schools and colleges, providing standard compound and stereo microscopes as well as cheap PC video based microscope solutions where the whole class can look at a computer screen with some pushing and shoving. Best to try them on 'approval' as many cheap microscopes can be disappointing if you expect too much.
Excellent pre-prepared stained slides of plant stems and leaves or bits of rats, insects etc.. can be bought, but they tend to be expensive and are easily broken by any age-group. Mounted slides keep well though, so 'vintage' ones even from 50 years ago can still look OK - most schools will have some specimen slides knocking about.
At home and our Primary school (under 11) we use the Digital Blue QX-5 (£70) - it's fun but pretty useless for serious microscope work as it's so low res (but at 640 x 480 better than the old Intel QX-3 it replaced). It puts the image on a PC screen. See http://micro.magnet.fsu.edu/optics/intelplay/index.html (not updated for the QX-5 but all applies - the site even discusses ways of contrast enhancement etc..). Once on the PC the 640x480 images can be manipulated and pasted etc, and the QX-5 does time-lapse for things like crystal growth. living plants growing and small animals. The similar but far better built Olympus MIC-D was great but being over £500 it was just too expensive for most schools and is now discontinued - there are other similar budget systems about though.
The macro on a good digital camera (like the image stabilised 5MP Canon S2IS going cheap at £200 over here - http://www.dpreview.com/reviews/canons2is) is also worth a try, particularly with a small tripod and halogen bendy desk lamp if very close-up, but I'm not sure I'd like a class with 20 boys near my Olympus E500 digital SLR system though. You can get quite reasonable pictures by resting a small compact digital camera lens against the eyepiece of a microscope.
By the way do try growing crystals on a slide, a few drops of a saturated solution of salt (NaCl) or copper sulphate will grow superb crystals on the surface of a slide when viewed under a microscope (but it takes a few hours for the crystals to form and they often look best before the liquids all gone). Just make sure they don't drive the objective tips into the solution. It's not biology but its fun.
Regards
Keith
Try looking in Amazon.com for decent microscope books with lots of pictures (and they have a good customer review system). Plus try web searches for general sites like these (and for more specific topics) :
http://www.101science.com/Microscope.htm http://micro.magnet.fsu.edu/ http://www.microscopy-uk.org.uk/index.html http://www.btinternet.com/~stephen.durr/ http://www.mccroneatlas.com http://www.diatoms.co.uk [for fun images made of diatoms]
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: gary-at-gaugler.com [mailto:gary-at-gaugler.com] Sent: 13 June 2006 05:12 To: keith.morris-at-ucl.ac.uk
This is an extremely broad and unbounded question. How much money are you willing to spend and what analysis methods do you want (LM, DF, DIC, PH, POL, etc.)?
I would recommend querying the usenet group about your question, as it comes up frequently.
sci.techniques.microscopy
Many Web apps have Usenet avenues. There is a large number of informed and experienced folks who deal with LMs on a daily basis. Many are quite helpful for non-professional applications. This is good since the opposite can get out of hand for some folks (i.e., too pricey).
gary g.
At 08:07 PM 6/12/2006, you wrote:
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==============================Original Headers============================== 12, 21 -- From gary-at-gaugler.com Mon Jun 12 23:09:20 2006 12, 21 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 12, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5D49KoL020603 12, 21 -- for {microscopy-at-microscopy.com} ; Mon, 12 Jun 2006 23:09:20 -0500 12, 21 -- Received: (qmail 22096 invoked from network); 12 Jun 2006 21:09:19 -0700 12, 21 -- Received: by simscan 1.1.0 ppid: 22093, pid: 22094, t: 0.1669s 12, 21 -- scanners: regex: 1.1.0 attach: 1.1.0 12, 21 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 12, 21 -- by qsmtp3 with SMTP; 12 Jun 2006 21:09:19 -0700 12, 21 -- Message-Id: {7.0.1.0.2.20060612210013.024ea218-at-gaugler.com} 12, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 12, 21 -- Date: Mon, 12 Jun 2006 21:09:21 -0700 12, 21 -- To: sullivanmassi-at-comcast.net 12, 21 -- From: Gary Gaugler {gary-at-gaugler.com} 12, 21 -- Subject: Re: [Microscopy] AskAMicroscopist: purchasing microscopes for 12, 21 -- high school biology 12, 21 -- Cc: MSA listserver {microscopy-at-microscopy.com} 12, 21 -- In-Reply-To: {200606130307.k5D37AWv001585-at-ns.microscopy.com} 12, 21 -- References: {200606130307.k5D37AWv001585-at-ns.microscopy.com} 12, 21 -- Mime-Version: 1.0 12, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
==============================Original Headers============================== 39, 27 -- From keith.morris-at-ucl.ac.uk Tue Jun 13 08:34:16 2006 39, 27 -- Received: from vscani-e2.ucl.ac.uk (vscani-e2.ucl.ac.uk [144.82.108.34]) 39, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DDYFDs002151 39, 27 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 08:34:16 -0500 39, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 39, 27 -- by vscani-e.ucl.ac.uk with esmtp (Exim 4.60) 39, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 39, 27 -- id 1Fq922-000671-Gd; Tue, 13 Jun 2006 14:34:10 +0100 39, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 39, 27 -- To: {sullivanmassi-at-comcast.net} 39, 27 -- Cc: {Microscopy-at-microscopy.com} 39, 27 -- Subject: RE: [Microscopy] Re: AskAMicroscopist: purchasing microscopes for 39, 27 -- Date: Tue, 13 Jun 2006 14:34:09 +0100 39, 27 -- Message-ID: {001b01c68eee$0d1cf500$7b865290-at-keithhigrade} 39, 27 -- MIME-Version: 1.0 39, 27 -- Content-Type: text/plain; 39, 27 -- charset="iso-8859-1" 39, 27 -- X-Mailer: Microsoft Office Outlook 11 39, 27 -- thread-index: AcaOn5P2KNX7omNpRVK9M3jp3oFQuwANfAkw 39, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 39, 27 -- In-Reply-To: {200606130412.k5D4CMXC030741-at-ns.microscopy.com} 39, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 39, 27 -- X-UCL-MailScanner: Found to be clean 39, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 39, 27 -- X-Spam-Status: No 39, 27 -- Content-Transfer-Encoding: 8bit 39, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5DDYFDs002151 ==============================End of - Headers==============================
On topic: The May issue of Microscopy Today contained the following joke:
"Scientifically, maybe body cells -do- replace themselves completely in seven years − but, legally, you’re still married."
Ron Anderson, MT Editor
nizets2-at-yahoo.com wrote: } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi, } } Probably the most interesting way to teach mitosis is } to synchronize cells in culture. It is probably not } the easiest though ;-). It is probable that } unsynchronized but very actively growing cells will } show enough mitotic events to cover the whole cycle. } Most of the time they are easy to recognize by normal } phase constrast, although fluorescent staining like } DAPI are very fast to perform and very impressive to } observe. } I must admit that after working now for more than 10 } years with cells in culture, seeing them split in late } telophase is still an exciting event. I witness events } which are actually occuring in my own body at that } very moment!!! } } I agree that you should insist on the functional part } though. It is not easy to comprehend these phenomenon } in themself, but understand the difference between } mitosis and meiosis is really a hard task. } } Good luck } } Stephane } } --- mhunt-at-flinnsci.com wrote: } } } } } ---------------------------------------------------------------------------- } } } The Microscopy ListServer -- CoSponsor: The } } Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.microscopy.com/MicroscopyListserver } } On-Line Help } } } } } http://www.microscopy.com/MicroscopyListserver/FAQ.html } } ---------------------------------------------------------------------------- } } } Below is the result of your feedback form } } (NJZFM-ultra-55). It was submitted by } } (mhunt-at-flinnsci.com) from } } } } } http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html } } } on Friday, May 26, 2006 at 09:59:18 } } } } } --------------------------------------------------------------------------- } } } Email: mhunt-at-flinnsci.com } } Name: Maureen Hunt } } } } Organization: Flinn } } } } Education: 9-12th Grade High School } } } } Location: Batavia, Illinois, USA } } } } Question: I am a former materials microscopist and } } teacher who is trying to find a more exciting way to } } teach mitosis and meiosis. Currently, everyone uses } } prepared slides of whitefish blastula to teach } } animal mitosis and either prepared slides or } } unprepared slides of allium root tips to teach plant } } mitosis. Is there a protist or other "animal-like" } } eukaryote that can be given to the students, } } squashed and stained to show animal mitosis? I have } } googled and come up short. Any suggestions? } } Thank-you for your time. } } } } } } } --------------------------------------------------------------------------- } } } } } ==============================Original } } Headers============================== } } 7, 13 -- From zaluzec-at-ultra5.microscopy.com Mon Jun } } 12 22:04:39 2006 } } 7, 13 -- Received: from [206.69.208.22] } } (mac22.zaluzec.com [206.69.208.22]) } } 7, 13 -- by ns.microscopy.com } } (8.12.11.20060308/8.12.8) with ESMTP id } } k5D34caY027718 } } 7, 13 -- for {microscopy-at-microscopy.com} ; Mon, 12 } } Jun 2006 22:04:38 -0500 } } 7, 13 -- Mime-Version: 1.0 } } 7, 13 -- X-Sender: zaluzec-at-ultra5.microscopy.com } } (Unverified) } } 7, 13 -- Message-Id: } } {p06110403c0b3db9f2108-at-[206.69.208.22]} } } 7, 13 -- Date: Mon, 12 Jun 2006 22:04:36 -0500 } } 7, 13 -- To: microscopy-at-microscopy.com } } 7, 13 -- From: mhunt-at-flinnsci.com (by way of } } Ask-A-Microscopist) } } 7, 13 -- Subject: AskAMicroscopist: trying to find } } a more exciting way to teach } } 7, 13 -- mitosis } } 7, 13 -- Content-Type: text/plain; } } charset="us-ascii" } } ==============================End of - } } Headers============================== } } } } } } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam } protection around } http://mail.yahoo.com } } __________________________________________________ } Do You Yahoo!? } Tired of spam? Yahoo! Mail has the best spam protection around } http://mail.yahoo.com } } ==============================Original Headers============================== } 10, 20 -- From nizets2-at-yahoo.com Tue Jun 13 03:02:15 2006 } 10, 20 -- Received: from web37413.mail.mud.yahoo.com (web37413.mail.mud.yahoo.com [209.191.87.66]) } 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5D82EIE028952 } 10, 20 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 03:02:15 -0500 } 10, 20 -- Received: (qmail 61561 invoked by uid 60001); 13 Jun 2006 08:02:14 -0000 } 10, 20 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 10, 20 -- s=s1024; d=yahoo.com; } 10, 20 -- h=Message-ID:Received:Date:From:Subject:To:Cc:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; } 10, 20 -- b=tOIkUDC8s+gOtnIjmKkJvWbDTGAQj+LFZnsv1ozdZzAzDVLXwfVMD/L2Wg73iyegOLD44ZKSqYVGjNScvzxwgYuXu9AWm8tLly+hjFfmeL0wkxd9mT+mpTwPjeXPGCz7M934lDezDdmiDR+9C3R1aYtmYl6GWYsoE0iyVEXRny4= ; } 10, 20 -- Message-ID: {20060613080214.61559.qmail-at-web37413.mail.mud.yahoo.com} } 10, 20 -- Received: from [80.122.101.102] by web37413.mail.mud.yahoo.com via HTTP; Tue, 13 Jun 2006 01:02:14 PDT } 10, 20 -- Date: Tue, 13 Jun 2006 01:02:14 -0700 (PDT) } 10, 20 -- From: Stephane Nizet {nizets2-at-yahoo.com} } 10, 20 -- Subject: Re: [Microscopy] AskAMicroscopist: trying to find a more exciting way to teach } 10, 20 -- To: mhunt-at-flinnsci.com } 10, 20 -- Cc: microscopy-at-microscopy.com } 10, 20 -- In-Reply-To: {200606130311.k5D3B2ck012854-at-ns.microscopy.com} } 10, 20 -- MIME-Version: 1.0 } 10, 20 -- Content-Type: text/plain; charset=iso-8859-1 } 10, 20 -- Content-Transfer-Encoding: 8bit } ==============================End of - Headers============================== } } }
==============================Original Headers============================== 5, 20 -- From randerson20-at-tampabay.rr.com Tue Jun 13 09:09:14 2006 5, 20 -- Received: from ms-smtp-01.tampabay.rr.com (ms-smtp-01.tampabay.rr.com [65.32.5.131]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DE9DBu012872 5, 20 -- for {Microscopy-at-Microscopy.Com} ; Tue, 13 Jun 2006 09:09:13 -0500 5, 20 -- Received: from [127.0.0.1] (rrcs-24-73-73-214.se.biz.rr.com [24.73.73.214]) 5, 20 -- by ms-smtp-01.tampabay.rr.com (8.13.6/8.13.6) with ESMTP id k5DE994f016559; 5, 20 -- Tue, 13 Jun 2006 10:09:12 -0400 (EDT) 5, 20 -- Message-ID: {448EC703.2040702-at-tampabay.rr.com} 5, 20 -- Date: Tue, 13 Jun 2006 10:09:07 -0400 5, 20 -- From: Ronald Anderson {randerson20-at-tampabay.rr.com} 5, 20 -- User-Agent: Thunderbird 1.5.0.4 (Windows/20060516) 5, 20 -- MIME-Version: 1.0 5, 20 -- To: nizets2-at-yahoo.com, Listserver {Microscopy-at-Microscopy.Com} 5, 20 -- Subject: Re: [Microscopy] Re: AskAMicroscopist: trying to find a more exciting 5, 20 -- way to teach 5, 20 -- References: {200606130802.k5D82UNl029179-at-ns.microscopy.com} 5, 20 -- In-Reply-To: {200606130802.k5D82UNl029179-at-ns.microscopy.com} 5, 20 -- Content-Type: text/plain; charset=UTF-8; format=flowed 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- X-Virus-Scanned: Symantec AntiVirus Scan Engine ==============================End of - Headers==============================
The Olympus with the EM camera is great for high frame rates. No laser. The spinning slits are not as confocal as spinning pinholes, but it is quite bright. I was recently torturing the EM camera and were getting rates that I did not think were possible. Check it out. David
On Jun 13, 2006, at 5:21 AM, iztok.dogsa-at-ijs.si wrote:
} } } } ---------------------------------------------------------------------- } ------ } The Microscopy ListServer -- CoSponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- http://www.microscopy.com/ } MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } ------ } } This Question/Comment was submitted to the Microscopy Listserver } using the WWW based Form at http://www.microscopy.com/MLFormMail.html } ---------------------------------------------------------------------- } ----- } Remember this posting is most likely not from a Subscriber, so when } replying } please copy both iztok.dogsa-at-ijs.si as well as the MIcroscopy } Listserver } ---------------------------------------------------------------------- } ----- } } Email: iztok.dogsa-at-ijs.si } Name: iztok dogsa } } Organization: IJS } } Title-Subject: [Filtered] confocal spinning disk microscope } } Question: Has anyone experince with confocal spinning disk microscopy? } } Which system would you suggest? } } We'd like to achieve really high frame rates, but don't want to buy } the laser. The arc lamp is Ok for us. } } What about BD CARV II imaging system? Is it better than olympus DSU? } } Thanks & regards to you all, } } Iztok } } } ---------------------------------------------------------------------- } ----- } } ==============================Original } Headers============================== } 12, 12 -- From zaluzec-at-microscopy.com Tue Jun 13 07:15:39 2006 } 12, 12 -- Received: from [206.69.208.22] (mac22.zaluzec.com } [206.69.208.22]) } 12, 12 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k5DCFcbR018836 } 12, 12 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 } 07:15:39 -0500 } 12, 12 -- Mime-Version: 1.0 } 12, 12 -- X-Sender: (Unverified) } 12, 12 -- Message-Id: {p0611040ec0b45c3f4728-at-[206.69.208.22]} } 12, 12 -- Date: Tue, 13 Jun 2006 07:15:37 -0500 } 12, 12 -- To: microscopy-at-microscopy.com } 12, 12 -- From: iztok.dogsa-at-ijs.si (by way of MicroscopyListserver) } 12, 12 -- Subject: viaWWW: confocal spinning disk microscope } 12, 12 -- Content-Type: text/plain; charset="us-ascii" } ==============================End of - } Headers==============================
==============================Original Headers============================== 6, 22 -- From Elliott-at-arizona.edu Tue Jun 13 09:58:05 2006 6, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DEw5Rp024151 6, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 09:58:05 -0500 6, 22 -- Received: from localhost (eomer.email.arizona.edu [10.0.0.219]) 6, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id A917BE400CD 6, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 07:58:04 -0700 (MST) 6, 22 -- Received: from [192.168.0.30] (doctorelliott.us [70.57.226.104]) 6, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id BBDDBE41125 6, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 07:58:00 -0700 (MST) 6, 22 -- Mime-Version: 1.0 (Apple Message framework v750) 6, 22 -- In-Reply-To: {200606131221.k5DCLHVB025321-at-ns.microscopy.com} 6, 22 -- References: {200606131221.k5DCLHVB025321-at-ns.microscopy.com} 6, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 6, 22 -- Message-Id: {FC930766-D51F-4A66-9CC4-F179FC978836-at-arizona.edu} 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- From: David Elliott {Elliott-at-arizona.edu} 6, 22 -- Subject: Re: [Microscopy] viaWWW: confocal spinning disk microscope 6, 22 -- Date: Tue, 13 Jun 2006 07:57:57 -0700 6, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 6, 22 -- X-Mailer: Apple Mail (2.750) 6, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
Thanks to everyone who responded to my question regarding sectioning carbon nanotubes. This listserve is a great resource.
Best wishes,
Soumitra
********************************************************************************** Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office) 505-646-3283 (lab) Fax: 505-646-3282 http://biology-web.nmsu.edu/ghoshroy/ghoshroy.htm http://emldata.nmsu.edu
==============================Original Headers============================== 5, 20 -- From sghoshro-at-nmsu.edu Tue Jun 13 10:02:51 2006 5, 20 -- Received: from cheech.nmsu.edu (cheech.nmsu.edu [128.123.34.14]) 5, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DF2oXd032025 5, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 10:02:51 -0500 5, 20 -- Received: (from nobody-at-localhost) 5, 20 -- by cheech.nmsu.edu (8.11.6/8.11.6) id k5DF2oI04183 5, 20 -- for Microscopy-at-microscopy.com; Tue, 13 Jun 2006 09:02:50 -0600 5, 20 -- Received: from 128.123.105.122 ( [128.123.105.122]) 5, 20 -- as user sghoshro-at-imap.nmsu.edu by webmail.nmsu.edu with HTTP; 5, 20 -- Tue, 13 Jun 2006 09:02:50 -0600 5, 20 -- Message-ID: {1150210970.448ed39a418dd-at-webmail.nmsu.edu} 5, 20 -- Date: Tue, 13 Jun 2006 09:02:50 -0600 5, 20 -- From: Soumitra Ghoshroy {sghoshro-at-nmsu.edu} 5, 20 -- To: Microscopy-at-microscopy.com 5, 20 -- Subject: sectioning carbon nanotubes 5, 20 -- MIME-Version: 1.0 5, 20 -- Content-Type: text/plain; charset=ISO-8859-1 5, 20 -- Content-Transfer-Encoding: 8bit 5, 20 -- User-Agent: Internet Messaging Program (IMP) 3.0 5, 20 -- X-Originating-IP: 128.123.105.122 ==============================End of - Headers==============================
I'm in the market for a stand alone, oil-less, glow discharge unit. Recommendations would be greatly appreciated.
Thanks, Kim
Kimberly A. Riddle Florida State University Biological Science Imaging Resource 119 Bio Unit I, 4370 Tallahassee, FL 32306 tel: 850.644.6519 fax: 850.644.0481 http://bsir.bio.fsu.edu
==============================Original Headers============================== 8, 21 -- From riddle-at-bio.fsu.edu Tue Jun 13 11:07:07 2006 8, 21 -- Received: from bio.fsu.edu (bio.fsu.edu [128.186.38.55]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DG77pA013652 8, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 11:07:07 -0500 8, 21 -- Received: from riddlepc.bio.fsu.edu (riddlepc.bio.fsu.edu [128.186.14.73]) 8, 21 -- by bio.fsu.edu (8.13.1/8.13.1) with ESMTP id k5DG2ioB085887 8, 21 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 8, 21 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 12:02:44 -0400 (EDT) 8, 21 -- (envelope-from riddle-at-bio.fsu.edu) 8, 21 -- Message-Id: {6.2.5.6.2.20060613120255.02985e88-at-bio.fsu.edu} 8, 21 -- X-Mailer: QUALCOMM Windows Eudora Version 6.2.5.6 8, 21 -- Date: Tue, 13 Jun 2006 12:06:10 -0400 8, 21 -- To: Microscopy-at-microscopy.com 8, 21 -- From: Kim Riddle {riddle-at-bio.fsu.edu} 8, 21 -- Subject: glow discharge units 8, 21 -- Mime-Version: 1.0 8, 21 -- Content-Type: text/plain; charset="us-ascii"; format=flowed 8, 21 -- X-FSU-Biology-MailScanner: Found to be clean 8, 21 -- X-FSU-Biology-MailScanner-SpamCheck: not spam, SpamAssassin (score=-4.399, 8, 21 -- required 8, autolearn=not spam, ALL_TRUSTED -1.80, BAYES_00 -2.60) 8, 21 -- X-FSU-Biology-MailScanner-From: riddle-at-bio.fsu.edu ==============================End of - Headers==============================
On Jun 13, 2006, at 9:07 AM, riddle-at-bio.fsu.edu wrote:
} I'm in the market for a stand alone, oil-less, glow discharge unit. } Recommendations would be greatly appreciated. } Dear Kim, We have used our Harrick Basic Plasma Cleaner for several trouble-free years, and they also make an Extended Plasma Cleaner, which is a larger model. You would have to buy an oil-free pump to go with either unit, but a diaphragm pump would be adequate for glow-discharging. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Tue Jun 13 13:49:31 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DInUSV004811 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 13:49:31 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by earth-ox-postvirus (Postfix) with ESMTP id 71257109A09 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 11:49:30 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id EE7CC109B2A 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 11:49:24 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200606131607.k5DG7FwU013823-at-ns.microscopy.com} 4, 22 -- References: {200606131607.k5DG7FwU013823-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {3476722cb03ade7cd1e8f052a6ab2203-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] glow discharge units 4, 22 -- Date: Tue, 13 Jun 2006 11:49:40 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
I have an individual learning TEM. He will be doing a lot of mouse brain tissue infected with HIV. What sort of safety or handling procedures should we follow? I don't have experience in this area with this sort of pathogen. The tissue is arriving in my lab fixed in 2%glutaraldehyde, 2%paraformaldehyde, 0.5%acrolein in .1M phosphate buffer which I provided to his lab. So, for processing up through embedding can we just handle the samples the same as any other tissue? When sectioning the blocks do the shavings from trimming the block, sectioning, etc. have to be specially collected? If he nicks himself with the razor blade while trimming the block is there an increased risk? Should I have him wearing exam gloves while trimming? Any and all advice will be greatly appreciated.
Tom Bargar Core Electron Microscopy Research Facility University of Nebraska Medical Center 986395 Nebraska Medical Center Omaha, NE 68198-6395 tbargar-at-unmc.edu 402-559-7347
==============================Original Headers============================== 6, 17 -- From tbargar-at-unmc.edu Tue Jun 13 13:56:53 2006 6, 17 -- Received: from zixvpm01.unmc.edu (zixvpm01.unmc.edu [192.198.54.126]) 6, 17 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DIurm7014950 6, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 13:56:53 -0500 6, 17 -- Received: from zixvpm01.unmc.edu (ZixVPM [127.0.0.1]) 6, 17 -- by Outbound.unmc.edu (Proprietary) with ESMTP id 1395C4C0CE 6, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 13:48:17 -0500 (CDT) 6, 17 -- Received: from unmcnotes01.unmc.edu (host-137-197-103-35.unmc.edu [137.197.103.35]) 6, 17 -- by zixvpm01.unmc.edu (Proprietary) with ESMTP id 07BB54C0C9 6, 17 -- for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 13 Jun 2006 13:48:16 -0500 (CDT) 6, 17 -- Subject: TEM of HIV in brain tissue 6, 17 -- To: Microscopy-at-MSA.Microscopy.Com 6, 17 -- Message-ID: {OF5A64AA61.19F4D29B-ON8625718C.0065E182-8625718C.006814A8-at-unmc.edu} 6, 17 -- From: Tom W Bargar {tbargar-at-unmc.edu} 6, 17 -- Date: Tue, 13 Jun 2006 13:56:50 -0500 6, 17 -- MIME-Version: 1.0 6, 17 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
Does anyone have a copy of a service manual for the Sorvall MT-2B microtomes? I have the (original) manuals that came with the things, but not a take-it-apart-and-fix-it service manual. If such things still exist, I would appreciate a photocopy. Many thanks!
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 2, 20 -- From oshel1pe-at-cmich.edu Tue Jun 13 15:08:08 2006 2, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 2, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DK88C8026809 2, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 15:08:08 -0500 2, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 2, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5DKh34l019524 2, 20 -- for {Microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 16:43:03 -0400 2, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 2, 20 -- Tue, 13 Jun 2006 16:08:07 -0400 2, 20 -- Mime-Version: 1.0 2, 20 -- Message-Id: {f06230913c0b4cb45f81f-at-[141.209.160.132]} 2, 20 -- Date: Tue, 13 Jun 2006 16:08:05 -0400 2, 20 -- To: Microscopy-at-microscopy.com 2, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 2, 20 -- Subject: Sorvall MT-2B service manuals? 2, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 2, 20 -- X-OriginalArrivalTime: 13 Jun 2006 20:08:08.0177 (UTC) FILETIME=[16908610:01C68F25] 2, 20 -- X-CanItPRO-Stream: default 2, 20 -- X-Spam-Score: -4 () L_EXCH_MF 2, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
Glutaraldehyde and Formaldehyde are both very good inactivators. When I do level 4 stuff they treat in 4%PF for 48hr to 4 weeks, depending on the protocol they follow at the time, so maybe you want to increase to a more Karnovsky like fix and make the PF 4%. That works for them, it should work for you. Under either condition, by the time you get your stuff it will be well inactivated, and safe.
Paul
==============================Original Headers============================== 4, 21 -- From paul_hazelton-at-umanitoba.ca Tue Jun 13 15:11:29 2006 4, 21 -- Received: from electra.cc.umanitoba.ca (electra.cc.umanitoba.ca [130.179.16.23]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5DKBSB0031349 4, 21 -- for {microscopy-at-microscopy.com} ; Tue, 13 Jun 2006 15:11:29 -0500 4, 21 -- Received: from umanitoba.ca (net40.medmb.umanitoba.ca [140.193.11.160]) 4, 21 -- (authenticated bits=0) 4, 21 -- by electra.cc.umanitoba.ca (8.13.0/8.13.0) with ESMTP id k5DKBQrf001617; 4, 21 -- Tue, 13 Jun 2006 15:11:26 -0500 (CDT) 4, 21 -- Message-ID: {448F1BEA.8000609-at-umanitoba.ca} 4, 21 -- Date: Tue, 13 Jun 2006 15:11:22 -0500 4, 21 -- From: paul r hazelton {paul_hazelton-at-umanitoba.ca} 4, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.1; en-US; rv:1.4) Gecko/20030624 Netscape/7.1 (ax) 4, 21 -- X-Accept-Language: en-us, en 4, 21 -- MIME-Version: 1.0 4, 21 -- To: tbargar-at-unmc.edu 4, 21 -- CC: Microscopy Listserver {microscopy-at-microscopy.com} 4, 21 -- Subject: Re: [Microscopy] TEM of HIV in brain tissue 4, 21 -- References: {200606131858.k5DIwHVW018669-at-ns.microscopy.com} 4, 21 -- In-Reply-To: {200606131858.k5DIwHVW018669-at-ns.microscopy.com} 4, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 4, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
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Question: Hi All, I was wondering if anyone call suggest a marker for endothelial cells in the rat brain (cryostat sections). I wish to count the density of blood vessels in the brains of rat pups who have been exposed to undernutrition or glucocorticoid treatment. I have tried CD31 from Chemicon with little success. I am looking for a structural marker of blood vessels- a marker which will stain all of the blood vessels and not be influenced or altered by undernutrition or hypoxia. Thanks in advance, Emily
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Email: Dandersen-at-optimedica.com Name: Dan Andersen
Organization: OptiMedica
Title-Subject: [Filtered] Histo Prep of ophthalmic samples for EM
Question: I am looking for a private laboraory to process some ocular samples I wish to analyze under EM.
On Jun 12, 2006, at 9:23 PM, Goinoutonalamb-at-yahoo.com wrote:
} Question: I am collaborating with a group on a 3D visualization } project through Brookhaven National Labs and I have a question } pertaining to 3D reconstruction by means of electron tomography. From } the literature that I have read, I've gathered that it's recommended } that a 200 kv beam or higher is used when working with sections of 120 } nm or thicker. Unfortunately, the philips EM-300 that we are working } with is only capable of producing a 100 kv beam. Because of the time } constraints of our project (10 weeks) it will be very difficult to } determine what section thicknesses/maximum tilt angles will work with } 100kv with trial and error. Is there a formula, or a guideline of } some sort to follow to determine how much mass a 100kv beam can pass } with minimal loss of resolution? We are planning on doing a tilt } series from -60 to +60 degrees, in 2 degree increments, at 120 nm } sections (hopefully). If anyone could lend me some insight as to } whether or not this is feasible, and how ! } far we can go, I would greatly appreciate it! } Dear Michael, Some issues not answered by the other person who responded to your post are what the specimen consists of, and what resolution you want. For biological materials and polymers consisting of low-Z materials, the penetration of the beam will be greater than for electronics components consisting of silicon or germanium, and minerals containing high-Z atoms will be more restricted still. On the other hand, some of the restriction on thickness comes from the fact that at high tilt, the effective thickness is greater than the specimen thickness--you mention 60 degrees, at which angle it is twice the thickness. You may be able to get significant information by taking stereo pairs; these are obtained at small tilts, usually {10 degrees, so a thicker specimen can be observed in 3-D through a stereo viewer, which may be all you need, and 3-D reconstructions can be made with such software as sterecon, which was used at the Wadsworth Center in Albany NY when I was there. If your specimen exceeds 120 nm by a small amount, you can still try tomography with an angular range less that +/-60 degrees and obtain useful data (although artifacts from the missing wedge will be more pronounced the smaller the angular range), and you can take dual-axis tilt series, which will reduce the missing wedge to a missing pyramid and lessen the missing data artifacts. Finally, if the Wadsworth Center is still a Regional Resource, you might be able to get time on one of their higher-voltage EMs, and they have a 1.2 MV scope, which should provide sufficient penetrating power even for roughly 1 um sections and/or for higher-Z materials. Good luck. Yours, Bill Tivol, PhD EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
==============================Original Headers============================== 4, 22 -- From tivol-at-caltech.edu Tue Jun 13 19:38:17 2006 4, 22 -- Received: from outgoing-mail.its.caltech.edu (outgoing-mail.its.caltech.edu [131.215.239.19]) 4, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5E0cGUs007753 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 19:38:17 -0500 4, 22 -- Received: from earth-dog.caltech.edu (earth-dog [192.168.1.3]) 4, 22 -- by fire-ox-postvirus (Postfix) with ESMTP id A8AF235624 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 17:38:15 -0700 (PDT) 4, 22 -- Received: from [192.168.157.234] (pix-1.caltech.edu [131.215.2.21]) 4, 22 -- by earth-ox.its.caltech.edu (Postfix) with ESMTP id C25E7109C6C 4, 22 -- for {microscopy-at-msa.microscopy.com} ; Tue, 13 Jun 2006 17:38:09 -0700 (PDT) 4, 22 -- Mime-Version: 1.0 (Apple Message framework v624) 4, 22 -- In-Reply-To: {200606130423.k5D4Nlk9004749-at-ns.microscopy.com} 4, 22 -- References: {200606130423.k5D4Nlk9004749-at-ns.microscopy.com} 4, 22 -- Content-Type: text/plain; charset=US-ASCII; format=flowed 4, 22 -- Message-Id: {fee7f3cf2eefaec0e4d6d3b0b3620142-at-caltech.edu} 4, 22 -- Content-Transfer-Encoding: 7bit 4, 22 -- From: Bill Tivol {tivol-at-caltech.edu} 4, 22 -- Subject: Re: [Microscopy] AskAMicroscopist: Tomographic Reconstruction 4, 22 -- Date: Tue, 13 Jun 2006 17:38:26 -0700 4, 22 -- To: microscopy-at-msa.microscopy.com 4, 22 -- X-Mailer: Apple Mail (2.624) 4, 22 -- X-Spam-Scanned: at Caltech-ITS on earth-dog by amavisd-2.3.3 ==============================End of - Headers==============================
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Email: aribaudo-at-triprinceton.org Name: Anthony Ribaudo
Organization: TRI Princeton
Title-Subject: [Filtered] TEM Sample Preparation of Paper using Foraldehyde
Question: Dear All,
Can anyone suggest the experimental setup, i.e. the apparatus needed to prepare paper samples that utilizes formaldehyde to crosslink the paper fibers. Formaldehyde imparts water-resistant qualities and increases the dimensional stability against moisture. The formaldehyde reaction conditions recommended are as follow: Temperature 140¢C. Reaction period 0.5-48 hours. Formaldehyde concentration introduced in vapor phase 2.19 x 10-2 mol/L.
After treatment the paper sample is embedded in epoxy resin and ultrasectioning.
Thank you,
Anthony Ribaudo TRI Princeton 601 Prospect Avenue Princeton, NJ 08542
I was asked to look into the morphology and cause of tin whiskers in high tin content lead free solders by a guy I meet. Some of the following questions have been asked for 50 years. Maybe someone out there can share some more recent expertise about tin whiskers.
* What is the root cause of crystalline tin whiskers? (It's probably not resolved yet. There are lots of conflicting opinions and observations on Google.)
* How do you prevent tin whiskers from growing out of almost pure tin solder joints? (He is not that interested in a thin film coating solution.) * What is the mechanism(s) that can cause 2 micron diameter or less "fibers" (et al) to grow right out of the surface of solder joins to a length of up to one observed to be 10 mm long? * What are the EDS results of tin whisker analysis? * Are the EDS analysis results settled science? * Have solder joints been ion milled to explore the interior of the solder or is this strictly a compression/stress surface phenomena? Or both?
He is NOT interested in high humidity dendritic growth of tin on PCB surfaces reputed to be caused by electric lines and associated equipotential lines. He is interested in the shorts caused by tin whiskers bridging solder joints through the air or a vacuum.
This raises the following big question. * How do you accelerate the growth of tin whiskers so that they can be studied without waiting 1-15 years for them to develop?
Please reply off list with any insights you have on this problem. I don't think this subject will have wide appeal. Thank you in advance for any perspectives that can be shared with him. If you are speculating on a solution or theory, please say so in your reply.
Paul
Disclaimers: I do not work for this guy's company, the guy, or any company or organization working on tin whiskers. I was astonished at the phenomenon, morphology, and how they ruined an obiting satellite. Tin whiskers grow in air or vacuum.
==============================Original Headers============================== 13, 24 -- From beaurega-at-westol.com Thu Jun 15 09:54:50 2006 13, 24 -- Received: from smtp-gateway-5.winbeam.com (smtp-gateway-5.winbeam.com [64.84.97.70]) 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5FEsnwQ003359 13, 24 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 09:54:50 -0500 13, 24 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) 13, 24 -- by smtp-gateway-5.winbeam.com (8.13.2/8.12.8) with SMTP id k5FEs1LD015079 13, 24 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 10:54:21 -0400 13, 24 -- Received: (qmail 30540 invoked by uid 89); 15 Jun 2006 14:53:29 -0000 13, 24 -- Received: from pitts-69-72-12-3.dynamic-dialup.coretel.net (HELO millenium) (69.72.12.3) 13, 24 -- by mail.winbeam.com with SMTP; 15 Jun 2006 14:53:29 -0000 13, 24 -- Message-Id: {3.0.6.32.20060615105431.007edaf0-at-pop3.norton.antivirus} 13, 24 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus 13, 24 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) 13, 24 -- Date: Thu, 15 Jun 2006 10:54:31 -0400 13, 24 -- To: microscopy-at-microscopy.com 13, 24 -- From: Beaurega {beaurega-at-westol.com} 13, 24 -- Subject: Tin Whiskers in high tin content solders. 13, 24 -- Mime-Version: 1.0 13, 24 -- Content-Type: text/plain; charset="us-ascii" 13, 24 -- X-Winbeam-MailScanner-Information: - Please contact Technical Support for more information 13, 24 -- X-Winbeam-MailScanner: Found to be clean [] (courtesy of Winbeam) 13, 24 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin (notcached, score=0, 13, 24 -- required 6, autolearn=disabled) 13, 24 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com ==============================End of - Headers==============================
You're right. There is not a clear concensus on the formation of tin whiskers. A couple of great sites on the subject are http://www.inemi.org/cms/newsroom/Presentations/TinWhiskerWorkshop_ECTC0 6.html and http://nepp.nasa.gov/whisker/
If you wander around on these sites, you'll find enough reports to read to keep you busy for quite a while. I notice that an April 2006 report from NASA ( http://nepp.nasa.gov/whisker/reference/tech_papers/2006-Leidecker-Tin-Wh isker-Failures.pdf ) states that "If you can't live with tin whiskers, then "Don't Use Tin"."
There are things that will keep whiskers from growing, most notably adding lead (but that probably doesn't help much).
Scratching the surface of the tin will influence more whisker growth (but I'm not sure if it'll be faster whisker growth). There are some extensive studies that have been done to try to find out what accelerates whisker growth, so I'd read those before starting my own study.
Hope this helps.
Diane _____________________________ Diane Ciaburri Principal Materials Engineer General Dynamics 100 Plastics Ave., Rm 2552A Pittsfield MA 01201 phone: (413) 494-3430 email: diane.ciaburri-at-gd-ais.com
-----Original Message----- X-from: beaurega-at-westol.com [mailto:beaurega-at-westol.com] Sent: Thursday, June 15, 2006 10:55 AM To: Ciaburri, Diane A.
Hello,
Anyone with contact info for a 3rd party service person/organization that can provide support services for a Rigaku Model: Ultra-18 XRD instrument, please contact Mr. Ross Potoff at the University of Arizona (520) 621-8101 or email to Potoff-at-email.arizona.edu.
Regards,
Bob Roberts EM Lab Services, Inc. 449 NW 62nd St. Topeka, Kansas 66617-1780 785.246.1232 voice 785.246.0168 fax www.emlabservices.com
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I would be curious to see any feedback on this subject, in the name of education.
I have a couple questions. Some of the lead-free solders have silver in them. Are you (or he) sure the whiskers are tin, or could they be silver? When you say "vacuum", are you referring to the low pressures in orbit, or a clean, high vacuum in a chamber of some sort? (or, perhaps, vacuum tube electronic component)
I believe the fields associated with a bias have something to do with it, but don't have any facts, or know what the mechanism is. I wonder if a high (strong) bias, in a vacuum chamber, would accelerate the phenomenon.
Looking forward to learning more. Thanks, Darrell
beaurega-at-westol.com wrote on 06/15/2006 10:55:59 AM:
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} } Hi, } } I was asked to look into the morphology and cause of tin whiskers in high } tin content lead free solders by a guy I meet. Some of the following } questions have been asked for 50 years. Maybe someone out there can share } some more recent expertise about tin whiskers. } } * What is the root cause of crystalline tin whiskers? (It's probably not } resolved yet. There are lots of conflicting opinions and observations on } Google.) } } * How do you prevent tin whiskers from growing out of almost pure tin } solder joints? } (He is not that interested in a thin film coating solution.) } * What is the mechanism(s) that can cause 2 micron diameter or less } "fibers" (et al) to grow right out of the surface of solder joins to a } length of up to one observed to be 10 mm long? } * What are the EDS results of tin whisker analysis? } * Are the EDS analysis results settled science? } * Have solder joints been ion milled to explore the interior of the solder } or is this strictly a compression/stress surface phenomena? Or both? } } He is NOT interested in high humidity dendritic growth of tin on PCB } surfaces reputed to be caused by electric lines and associated } equipotential lines. He is interested in the shorts caused by tin whiskers } bridging solder joints through the air or a vacuum. } } This raises the following big question. } * How do you accelerate the growth of tin whiskers so that they can be } studied without waiting 1-15 years for them to develop? } } Please reply off list with any insights you have on this problem. I don't } think this subject will have wide appeal. Thank you in advance for any } perspectives that can be shared with him. } If you are speculating on a solution or theory, please say so in your reply. } } Paul } } Disclaimers: I do not work for this guy's company, the guy, or any company } or organization working on tin whiskers. } I was astonished at the phenomenon, morphology, and how they ruined an } obiting satellite. } Tin whiskers grow in air or vacuum. } } } } } } } ==============================Original Headers============================== } 13, 24 -- From beaurega-at-westol.com Thu Jun 15 09:54:50 2006 } 13, 24 -- Received: from smtp-gateway-5.winbeam.com (smtp-gateway-5. } winbeam.com [64.84.97.70]) } 13, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with } ESMTP id k5FEsnwQ003359 } 13, 24 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 09:54:50 -0500 } 13, 24 -- Received: from mail.winbeam.com (mail.winbeam.com [64.84.96.10]) } 13, 24 -- by smtp-gateway-5.winbeam.com (8.13.2/8.12.8) with SMTP } id k5FEs1LD015079 } 13, 24 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 10:54:21 -0400 } 13, 24 -- Received: (qmail 30540 invoked by uid 89); 15 Jun 2006 14: } 53:29 -0000 } 13, 24 -- Received: from pitts-69-72-12-3.dynamic-dialup.coretel.net } (HELO millenium) (69.72.12.3) } 13, 24 -- by mail.winbeam.com with SMTP; 15 Jun 2006 14:53:29 -0000 } 13, 24 -- Message-Id: {3.0.6.32.20060615105431.007 edaf0-at-pop3.norton.antivirus} } 13, 24 -- X-Sender: beaurega/mail.westol.com-at-pop3.norton.antivirus } 13, 24 -- X-Mailer: QUALCOMM Windows Eudora Light Version 3.0.6 (32) } 13, 24 -- Date: Thu, 15 Jun 2006 10:54:31 -0400 } 13, 24 -- To: microscopy-at-microscopy.com } 13, 24 -- From: Beaurega {beaurega-at-westol.com} } 13, 24 -- Subject: Tin Whiskers in high tin content solders. } 13, 24 -- Mime-Version: 1.0 } 13, 24 -- Content-Type: text/plain; charset="us-ascii" } 13, 24 -- X-Winbeam-MailScanner-Information: - Please contact } Technical Support for more information } 13, 24 -- X-Winbeam-MailScanner: Found to be clean [] (courtesy of Winbeam) } 13, 24 -- X-Winbeam-MailScanner-SpamCheck: notspam, spamassassin } (notcached, score=0, } 13, 24 -- required 6, autolearn=disabled) } 13, 24 -- X-Winbeam-MailScanner-From: beaurega-at-westol.com } ==============================End of - Headers==============================
==============================Original Headers============================== 10, 29 -- From milesd-at-us.ibm.com Thu Jun 15 15:33:05 2006 10, 29 -- Received: from e3.ny.us.ibm.com (e3.ny.us.ibm.com [32.97.182.143]) 10, 29 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5FKX5Xm010224 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 15:33:05 -0500 10, 29 -- Received: from d01relay04.pok.ibm.com (d01relay04.pok.ibm.com [9.56.227.236]) 10, 29 -- by e3.ny.us.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5FKX3tL022193 10, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=OK) 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 16:33:03 -0400 10, 29 -- Received: from d01av02.pok.ibm.com (d01av02.pok.ibm.com [9.56.224.216]) 10, 29 -- by d01relay04.pok.ibm.com (8.13.6/NCO/VER7.0) with ESMTP id k5FKX1ld264774 10, 29 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 16:33:03 -0400 10, 29 -- Received: from d01av02.pok.ibm.com (loopback [127.0.0.1]) 10, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.13.3) with ESMTP id k5FKX1nS002256 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 16:33:01 -0400 10, 29 -- Received: from d01ml076.pok.ibm.com (d01ml076.pok.ibm.com [9.56.229.50]) 10, 29 -- by d01av02.pok.ibm.com (8.12.11.20060308/8.12.11) with ESMTP id k5FKX1Ll002235 10, 29 -- for {Microscopy-at-MSA.Microscopy.Com} ; Thu, 15 Jun 2006 16:33:01 -0400 10, 29 -- In-Reply-To: {200606151455.k5FEtxZZ004881-at-ns.microscopy.com} 10, 29 -- Subject: Re: [Microscopy] Tin Whiskers in high tin content solders. 10, 29 -- To: Microscopy-at-MSA.Microscopy.Com 10, 29 -- X-Mailer: Lotus Notes Release 7.0 HF85 November 04, 2005 10, 29 -- Message-ID: {OF7455FAA7.1B918C5E-ON8525718E.006F3FB0-8525718E.0070E1DC-at-us.ibm.com} 10, 29 -- From: Darrell Miles {milesd-at-us.ibm.com} 10, 29 -- Date: Thu, 15 Jun 2006 16:32:59 -0400 10, 29 -- X-MIMETrack: Serialize by Router on D01ML076/01/M/IBM(Release 7.0.1 HF4|May 16, 2006) at 10, 29 -- 06/15/2006 16:33:00 10, 29 -- MIME-Version: 1.0 10, 29 -- Content-type: text/plain; charset=US-ASCII ==============================End of - Headers==============================
The whiskers are tin. Silver is added to lead solder to reduce the melting point.
Whiskering occurs at room temperature, high and low humidity and all sorts of conditions. It is a nasty problem for commercial and especially MIL applications. Alternate solders are being studied (different combinations of elements) but their melting point is sufficiently higher than Pb solder to be a real problem.
gary g.
At 01:35 PM 6/15/2006, you wrote:
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==============================Original Headers============================== 10, 20 -- From gary-at-gaugler.com Thu Jun 15 16:10:07 2006 10, 20 -- Received: from qsmtp3.mc.surewest.net (qsmtp.mc.surewest.net [66.60.130.145]) 10, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5FLA6D2021040 10, 20 -- for {microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 16:10:06 -0500 10, 20 -- Received: (qmail 5097 invoked from network); 15 Jun 2006 14:10:06 -0700 10, 20 -- Received: by simscan 1.1.0 ppid: 5093, pid: 5094, t: 0.2385s 10, 20 -- scanners: regex: 1.1.0 attach: 1.1.0 10, 20 -- Received: from unknown (HELO ivan.gaugler.com) (66.60.171.211) 10, 20 -- by qsmtp3 with SMTP; 15 Jun 2006 14:10:05 -0700 10, 20 -- Message-Id: {7.0.1.0.2.20060615140636.024fa7d0-at-gaugler.com} 10, 20 -- X-Mailer: QUALCOMM Windows Eudora Version 7.0.1.0 10, 20 -- Date: Thu, 15 Jun 2006 14:10:06 -0700 10, 20 -- To: milesd-at-us.ibm.com 10, 20 -- From: Gary Gaugler {gary-at-gaugler.com} 10, 20 -- Subject: Re: [Microscopy] Re: Tin Whiskers in high tin content solders. 10, 20 -- Cc: MSA listserver {microscopy-at-microscopy.com} 10, 20 -- In-Reply-To: {200606152035.k5FKZcD3013387-at-ns.microscopy.com} 10, 20 -- References: {200606152035.k5FKZcD3013387-at-ns.microscopy.com} 10, 20 -- Mime-Version: 1.0 10, 20 -- Content-Type: text/plain; charset="us-ascii"; format=flowed ==============================End of - Headers==============================
I am in the middle of efforts to transfer manufacturing operations from Lead:Tin solder to Lead Free solder. Presently Silver and copper are used with the tin to reduce the melting point to about 220C. There are many different theories about why tin whiskers form including a compressive stress on the tin layer. The stress may be caused by copper diffusing into the tin at grain boundaries and forming copper:tin complexes which take up more volume.
There is a historical paper covering the history of the research entitled: A History of Tin Whisker Theory: 1946 to 2004 George T. Galyon IBM eSG Group Poughkeepsie, New York
I'm sorry I can't find the original source for the article. It isn't mentioned on my document. But George T. Gaylon has been (and may still be) associated with iNEMI. Some of his material is available through them at http://www.inemi.org/cms/newsroom/NEMI_NIST_TMS_workshop.html
Hope this helps. The electronics industry has done a lot of work in this area in the past five or so years.
Mark Woolley PTRL Lab Avaya, Inc. 1300 West 120th Ave. Westminster, CO 80234 (303) 538-9179 Office (303) 538-2166 Lab woolleym-at-avaya.com
==============================Original Headers============================== 8, 24 -- From woolleym-at-avaya.com Thu Jun 15 18:21:15 2006 8, 24 -- Received: from co300216-ier2.net.avaya.com (co300216-ier2.net.avaya.com [198.152.13.103]) 8, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5FNLFaH006392 8, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 18:21:15 -0500 8, 24 -- Received: from cof110avexu2.global.avaya.com (h135-9-6-17.avaya.com [135.9.6.17]) 8, 24 -- by co300216-ier2.net.avaya.com (Switch-3.1.8/Switch-3.1.7) with ESMTP id k5FNIbcK023470 8, 24 -- for {Microscopy-at-microscopy.com} ; Thu, 15 Jun 2006 19:18:38 -0400 8, 24 -- X-MimeOLE: Produced By Microsoft Exchange V6.0.6603.0 8, 24 -- content-class: urn:content-classes:message 8, 24 -- MIME-Version: 1.0 8, 24 -- Content-Type: text/plain; 8, 24 -- charset="iso-8859-1" 8, 24 -- Subject: Tin Whiskers 8, 24 -- Date: Thu, 15 Jun 2006 17:21:14 -0600 8, 24 -- Message-ID: {A49F94D2EEDFB74E816FF4277B7A602C01BF13A2-at-cof110avexu2.global.avaya.com} 8, 24 -- X-MS-Has-Attach: 8, 24 -- X-MS-TNEF-Correlator: 8, 24 -- Thread-Topic: Tin Whiskers 8, 24 -- Thread-Index: AcaQ0mWIOWa4wn+1TAy0Ej7r8a8LEA== 8, 24 -- From: "Woolley, Mark D \(Mark\)" {woolleym-at-avaya.com} 8, 24 -- To: {Microscopy-at-microscopy.com} 8, 24 -- X-Scanner: InterScan AntiVirus for Sendmail 8, 24 -- Content-Transfer-Encoding: 8bit 8, 24 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5FNLFaH006392 ==============================End of - Headers==============================
Hi all. I would like to take a look a the moth eye structure or a fly, but I'm completely new to biological sample... What kind of sample preparation is recommended (any essiccation procedure)? Or it is simple possible to coat it with a metal layer and this will avoid it to explode and splatter around the column? what will happen in the latter case? (just will have to wait longer for a good vacuum next time, or ppermanent damage to any parts?)
Please let me know. Any help is welcome. All the best, Marco.
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx Dr. Marco Salerno, Ph.D.: marco.salerno-at-unile.it CNR-INFM Center NNL: www.nnl.it c/o Palazzine Garrisi, via per Arnesano 16 (km 5) 73100 Lecce - Italy Off. +39-0832-29.8236 Lab.+39-0832-29.8336/8347 Mob.+39-349-26.75.277 Fax +39-0832-29.8238 Home +39-0832-32.51.45 xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
==============================Original Headers============================== 6, 22 -- From marco.salerno-at-unile.it Fri Jun 16 02:03:27 2006 6, 22 -- Received: from mailing.unile.it (mailing.unile.it [193.204.77.251]) 6, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5G73QZp026611 6, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 02:03:27 -0500 6, 22 -- Received: from MSALERNO (unknown [193.204.74.205]) 6, 22 -- by mailing.unile.it (Postfix) with SMTP id 04BE8104B71 6, 22 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 09:03:25 +0200 (CEST) 6, 22 -- Message-ID: {029d01c69113$067b2e20$cc1e0a0a-at-Utente} 6, 22 -- From: "Marco Salerno" {marco.salerno-at-unile.it} 6, 22 -- To: {Microscopy-at-microscopy.com} 6, 22 -- Subject: first time of a soft matter specimen, please help! 6, 22 -- Date: Fri, 16 Jun 2006 09:03:52 +0200 6, 22 -- MIME-Version: 1.0 6, 22 -- Content-Type: text/plain; 6, 22 -- format=flowed; 6, 22 -- charset="iso-8859-1"; 6, 22 -- reply-type=original 6, 22 -- Content-Transfer-Encoding: 7bit 6, 22 -- X-Priority: 3 6, 22 -- X-MSMail-Priority: Normal 6, 22 -- X-Mailer: Microsoft Outlook Express 6.00.2900.2869 6, 22 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 ==============================End of - Headers==============================
you will find some EM analytical labs at this site: http://microanalysis.mikroanalytik.de/service%20microanalysis%20e.phtml
Regards
Frank
Dandersen-at-optimedica.com wrote:
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==============================Original Headers============================== 7, 21 -- From eggert-at-mikroanalytik.de Fri Jun 16 04:48:19 2006 7, 21 -- Received: from Mail1.KONTENT.De (mail1.kontent.de [81.88.34.36]) 7, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5G9mIxG008591 7, 21 -- for {microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 04:48:18 -0500 7, 21 -- Received: from mikroanalytik.de (p54BDF1C1.dip.t-dialin.net [84.189.241.193]) 7, 21 -- by Mail1.KONTENT.De (Postfix) with ESMTP id 858DE104A3F2; 7, 21 -- Fri, 16 Jun 2006 11:48:19 +0200 (CEST) 7, 21 -- Message-ID: {44927F30.4060702-at-mikroanalytik.de} 7, 21 -- Date: Fri, 16 Jun 2006 11:51:44 +0200 7, 21 -- From: Frank Eggert {eggert-at-mikroanalytik.de} 7, 21 -- User-Agent: Mozilla/5.0 (Windows; U; Windows NT 5.0; de-DE; rv:1.4) Gecko/20030619 Netscape/7.1 (ax) 7, 21 -- X-Accept-Language: de, de-de, en 7, 21 -- MIME-Version: 1.0 7, 21 -- To: microscopy-at-microscopy.com 7, 21 -- Cc: Dandersen-at-optimedica.com 7, 21 -- Subject: Re: [Microscopy] viaWWW: private laboraory to process some ocular 7, 21 -- samples 7, 21 -- References: {200606140030.k5E0Um81027205-at-ns.microscopy.com} 7, 21 -- In-Reply-To: {200606140030.k5E0Um81027205-at-ns.microscopy.com} 7, 21 -- Content-Type: text/plain; charset=ISO-8859-1; format=flowed 7, 21 -- Content-Transfer-Encoding: 7bit ==============================End of - Headers==============================
I routinely image insects in the SEM without any preparation other than sputter coating. I kill the insects first with ethyl acetate fumes or CO2. Then mount using a double-sticky carbon conductive tab, and coat. Note: if the insect/etc. only contacts the mounting tab in a small area because of convexity, you may have to also use some silver paint to improve the electrical ground. This is commonly needed (something I do automatically). The paint requires a wait to dry. Nor is it always needed -- the University president brought his 6 year-old son to our facility with a beetle to see the SEM. It was a quick mount, no paint, 3 minute Au coat, and in the SEM. Good points for the facility, too. If the critters aren't particularly hairy or you have access to an ESEM, you might be able to leave off the coating. Some insects and arachnids can be imaged at low kVs without coating, and will survive the experience. Note though, that soft-bodied arthropods can (and will) shrink and distort in the vacuum. Robust ones hold up better. Beetles are particularly good for this. Make certain your insect is *intact*. Any missing limbs, antennae, etc. will allow for haemolymph to be sucked out into the chamber. This will cause the body to collapse and coat the innards of your 'scope with bug goo. Not good. I wouldn't do this at all in an instrument used for high-resolution or analytical work. Personally, though, external eye structure is a bit boring. Feet and especially mouthparts are much more interesting. Plus, their morphology is directly related to ecology. Eyes are, yes, but less obviously in external SEM-level anatomy. There are lots of examples on the web, as well as books of example images, with technique chapters.
Phil
} Hi all. } I would like to take a look } a the moth eye structure or a fly, } but I'm completely new to } biological sample... } What kind of sample preparation is recommended } (any essiccation procedure)? } Or it is simple possible to coat it with a } metal layer and this will avoid it to explode and splatter } around the column? } what will happen in the latter case? } (just will have to wait longer for a good vacuum } next time, or ppermanent damage to any parts?) } } Please let me know. } Any help is welcome. } All the best, } Marco. } } } xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx } Dr. Marco Salerno, Ph.D.: } marco.salerno-at-unile.it } CNR-INFM Center NNL: } www.nnl.it } c/o Palazzine Garrisi, } via per Arnesano 16 (km 5) } 73100 Lecce - Italy } Off. +39-0832-29.8236 } Lab.+39-0832-29.8336/8347 } Mob.+39-349-26.75.277 } Fax +39-0832-29.8238 } Home +39-0832-32.51.45 } xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 4, 24 -- From oshel1pe-at-cmich.edu Fri Jun 16 07:40:12 2006 4, 24 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 4, 24 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GCeCvW022096 4, 24 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 07:40:12 -0500 4, 24 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 4, 24 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5GDEx4l012931; 4, 24 -- Fri, 16 Jun 2006 09:14:59 -0400 4, 24 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 4, 24 -- Fri, 16 Jun 2006 08:40:09 -0400 4, 24 -- Mime-Version: 1.0 4, 24 -- Message-Id: {f06230903c0b851ca1e3b-at-[141.209.160.132]} 4, 24 -- In-Reply-To: {200606160710.k5G7AHYu001950-at-ns.microscopy.com} 4, 24 -- References: {200606160710.k5G7AHYu001950-at-ns.microscopy.com} 4, 24 -- Date: Fri, 16 Jun 2006 08:40:06 -0400 4, 24 -- To: marco.salerno-at-unile.it 4, 24 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 4, 24 -- Subject: Re: [Microscopy] first time of a soft matter specimen, please 4, 24 -- help! 4, 24 -- Cc: Microscopy-at-microscopy.com 4, 24 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 4, 24 -- X-OriginalArrivalTime: 16 Jun 2006 12:40:10.0015 (UTC) FILETIME=[012BBAF0:01C69142] 4, 24 -- X-CanItPRO-Stream: default 4, 24 -- X-Spam-Score: -4 () L_EXCH_MF 4, 24 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both mbisher-at-princeton.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: mbisher-at-princeton.edu Name: Margaret Bisher
Organization: Princeton University
Title-Subject: [Filtered] TEM of Soil Microbes
Question: Over the summer we offer a short course for high school science teachers in our department (Molecular Biology). Something the instructors would like to incorporate this year, would be to look at, if possible, their samples in the TEM. We did try it last year and met with limited sucess so I thought I would post this question to the group before this year's class. I am wondering if there is a method for them to clean up their "dirt" to get a better yield (I did find some phage and bacteria but I had to spend hours looking, it would have more exciting for them if there been more to look at) and then I can possibly make a suspension and then do negative stain on the sample, to see if we have any bacteria, etc. in the sample.
I'd appreciate any suggestions you might be able to supply.
Thanks, Peggy Bisher
Electron Microscopy & Histology Core Facility Manager Princeton University Department of Molecular Biology Moffett Laboratory, Room 113B Washington Road Princeton, New Jersey 08544 Voice: (609) 258-7026 Fax: (609) 258-8468 Email: mbisher-at-princeton.edu
If this phenomenon can be reliably reproduced, the effect from bias should be easily confirmed or denied.
This appears to be some type of transport mechanism... vapor transport, surface transport, bulk diffusion from the body of the metal, or grain boundary diffusion. It would be interesting to find where tin is being depleted in the solder joint, perhaps using EDS methods and metallographic cross sections. The challenge would be to distinguish tin depletion from normal solder casting segregation.
At the moment I'm assuming the whisker is a single crystal. Perhaps the whisker can be studied using x-ray crystallographic methods - either Laue or TEM methods. This may give a clue to the growth mechanism.
Stu Smalinskas, P.E. Metallurgist SKF Plymouth, Michigan
--- gary-at-gaugler.com wrote:
The whiskers are tin. Silver is added to lead solder to reduce the melting point.
Whiskering occurs at room temperature, high and low humidity and all sorts of conditions. It is a nasty problem for commercial and especially MIL applications. Alternate solders are being studied (different combinations of elements) but their melting point is sufficiently higher than Pb solder to be a real problem.
gary g.
---------------------------------------
At 01:35 PM 6/15/2006, you wrote:
Hi,
I would be curious to see any feedback on this subject, in the name of education.
I have a couple questions. Some of the lead-free solders have silver in them. Are you (or he) sure the whiskers are tin, or could they be silver? When you say "vacuum", are you referring to the low pressures in orbit, or a clean, high vacuum in a chamber of some sort? (or, perhaps, vacuum tube electronic component)
I believe the fields associated with a bias have something to do with it, but don't have any facts, or know what the mechanism is. I wonder if a high (strong) bias, in a vacuum chamber, would accelerate the phenomenon.
Looking forward to learning more. Thanks, Darrell
-----------------------------------------------
beaurega-at-westol.com wrote on 06/15/2006 10:55:59 AM:
Hi, I was asked to look into the morphology and cause of tin whiskers in high tin content lead free solders by a guy I meet. Some of the following questions have been asked for 50 years. Maybe someone out there can share some more recent expertise about tin whiskers.
* What is the root cause of crystalline tin whiskers? (It's probably not resolved yet. There are lots of conflicting opinions and observations on Google.)
* How do you prevent tin whiskers from growing out of almost pure tin solder joints? (He is not that interested in a thin film coating solution.)
* What is the mechanism(s) that can cause 2 micron diameter or less "fibers" (et al) to grow right out of the surface of solder joins to a length of up to one observed to be 10 mm long?
* What are the EDS results of tin whisker analysis?
* Are the EDS analysis results settled science?
* Have solder joints been ion milled to explore the interior of the solder or is this strictly a compression/stress surface phenomena? Or both?
He is NOT interested in high humidity dendritic growth of tin on PCB surfaces reputed to be caused by electric lines and associated equipotential lines. He is interested in the shorts caused by tin whiskers bridging solder joints through the air or a vacuum.
This raises the following big question. * How do you accelerate the growth of tin whiskers so that they can be studied without waiting 1-15 years for them to develop?
Please reply off list with any insights you have on this problem. I don't think this subject will have wide appeal. Thank you in advance for any perspectives that can be shared with him. If you are speculating on a solution or theory, please say so in your reply.
Paul
Disclaimers: I do not work for this guy's company, the guy, or any company or organization working on tin whiskers. I was astonished at the phenomenon, morphology, and how they ruined an obiting satellite. Tin whiskers grow in air or vacuum.
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 32, 19 -- From smalinskas-at-yahoo.com Fri Jun 16 09:53:13 2006 32, 19 -- Received: from web34410.mail.mud.yahoo.com (web34410.mail.mud.yahoo.com [66.163.178.159]) 32, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5GErDcf012840 32, 19 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 16 Jun 2006 09:53:13 -0500 32, 19 -- Received: (qmail 66812 invoked by uid 60001); 16 Jun 2006 14:53:12 -0000 32, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 32, 19 -- s=s1024; d=yahoo.com; 32, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Content-Type:Content-Transfer-Encoding; 32, 19 -- b=ZaqnxzqR/WQm9UGzGSDQ9QDR2u7EqRGDp1gUTTsmNfHkKxWlQH1BwTvFTXVYpBXKX4SmcKwCyVc8BUbFNLgWPbdC7Ww1zCaT+83IV0/q39xwxFYWr3jIz33E5TV99VdjgCAJSXnlo9AnVLNU6uYPnQKuaZuNjlylNT6ruPENBMQ= ; 32, 19 -- Message-ID: {20060616145312.66810.qmail-at-web34410.mail.mud.yahoo.com} 32, 19 -- Received: from [141.151.33.213] by web34410.mail.mud.yahoo.com via HTTP; Fri, 16 Jun 2006 07:53:12 PDT 32, 19 -- Date: Fri, 16 Jun 2006 07:53:12 -0700 (PDT) 32, 19 -- From: Kestutis Smalinskas {smalinskas-at-yahoo.com} 32, 19 -- Subject: Re: [Microscopy] Tin Whiskers in high tin content solders. 32, 19 -- To: microscopy-at-ns.microscopy.com 32, 19 -- In-Reply-To: {200606152112.k5FLCRUK023781-at-ns.microscopy.com} 32, 19 -- MIME-Version: 1.0 32, 19 -- Content-Type: text/plain; charset=iso-8859-1 32, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
I recall that there is a MT-2B in rm 110. What I don't remember is whether or not you have anywhere a *service* manual for one. We have the manuals that were shipped with the microtomes, even one old enough to be in a binder. We don't have a service manual. Do you have one, and if yes, can I get a photocopy of it? We have 3 of them, one horrible that's at best parts, and one I'm trying to resurrect before classes start this fall. Thanks.
Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 3, 20 -- From oshel1pe-at-cmich.edu Fri Jun 16 12:34:22 2006 3, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 3, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GHYMD1026600 3, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 12:34:22 -0500 3, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 3, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5GI9B4l003317 3, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 14:09:11 -0400 3, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 3, 20 -- Fri, 16 Jun 2006 13:34:22 -0400 3, 20 -- Mime-Version: 1.0 3, 20 -- Message-Id: {f0623090fc0b89b645ec8-at-[141.209.160.132]} 3, 20 -- Date: Fri, 16 Jun 2006 13:34:20 -0400 3, 20 -- To: Microscopy-at-microscopy.com 3, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 3, 20 -- Subject: MT-2B 3, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 3, 20 -- X-OriginalArrivalTime: 16 Jun 2006 17:34:22.0477 (UTC) FILETIME=[1ADBABD0:01C6916B] 3, 20 -- X-CanItPRO-Stream: default 3, 20 -- X-Spam-Score: -4 () L_EXCH_MF 3, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
If anyone out there has been doing any immunogold labeling for GFP, I would be interested in what primary you're using and how well it labels, depending on fixation.
Have a good weekend....
Lesley
Lesley S. Bechtold Senior Manager, Histopathology & Microscopy Sciences The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6322
==============================Original Headers============================== 8, 21 -- From lesley.bechtold-at-jax.org Fri Jun 16 12:47:10 2006 8, 21 -- Received: from carmen.jax.org (carmen.jax.org [192.43.249.16]) 8, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GHlAhu004528 8, 21 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 16 Jun 2006 12:47:10 -0500 8, 21 -- Received: from jcs-mid-prod.jax.org (jcs-mid-prod.jax.org [192.43.249.134]) 8, 21 -- by carmen.jax.org (8.12.11/8.12.11) with ESMTP id k5GHP7ph004720 8, 21 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 16 Jun 2006 13:47:08 -0400 (EDT) 8, 21 -- (envelope-from lesley.bechtold-at-jax.org) 8, 21 -- Received: from spikey.jax.org by jcs-mid-prod.jax.org 8, 21 -- with ESMTP id 99719541150479984; Fri, 16 Jun 2006 13:46:24 -0400 8, 21 -- From: "Lesley Bechtold" {lesley.bechtold-at-jax.org} 8, 21 -- To: "Microscopy Network" {microscopy-at-ns.microscopy.com} 8, 21 -- Subject: Immunogold Labeling for GFP 8, 21 -- Date: Fri, 16 Jun 2006 13:46:23 -0400 8, 21 -- Message-ID: {20060616134623491.00000004792-at-spikey} 8, 21 -- X-Mailer: Oracle Connector for Outlook 10.1.1.0.5 71011 (11.0.8010) 8, 21 -- X-Accept-Language: en-us, en 8, 21 -- MIME-Version: 1.0 8, 21 -- Content-Type: text/plain; charset=us-ascii 8, 21 -- Content-Transfer-Encoding: 8bit 8, 21 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5GHlAhu004528 ==============================End of - Headers==============================
Sorry. My mousing finger glitched and picked the wrong name in my address book. Phil -- Philip Oshel Microscopy Facility Supervisor Department of Biology Central Michigan University 024C Brooks Hall Mt. Pleasant, MI 48859 (989) 774-3576
==============================Original Headers============================== 1, 20 -- From oshel1pe-at-cmich.edu Fri Jun 16 13:30:17 2006 1, 20 -- Received: from ob2.cmich.edu (ob2.cmich.edu [141.209.20.21]) 1, 20 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GIUHA5015534 1, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 13:30:17 -0500 1, 20 -- Received: from egatea.central.cmich.local ([141.209.15.74]) 1, 20 -- by ob2.cmich.edu (8.12.10/8.12.10) with ESMTP id k5GJ554l007634 1, 20 -- for {Microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 15:05:05 -0400 1, 20 -- Received: from [141.209.160.132] ([141.209.160.132]) by egatea.central.cmich.local with Microsoft SMTPSVC(6.0.3790.1830); 1, 20 -- Fri, 16 Jun 2006 14:30:16 -0400 1, 20 -- Mime-Version: 1.0 1, 20 -- Message-Id: {f06230914c0b8a9028fd5-at-[141.209.160.132]} 1, 20 -- Date: Fri, 16 Jun 2006 14:30:14 -0400 1, 20 -- To: Microscopy-at-microscopy.com 1, 20 -- From: Philip Oshel {oshel1pe-at-cmich.edu} 1, 20 -- Subject: Oops MT-2B 1, 20 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed" 1, 20 -- X-OriginalArrivalTime: 16 Jun 2006 18:30:16.0791 (UTC) FILETIME=[EA2F5E70:01C69172] 1, 20 -- X-CanItPRO-Stream: default 1, 20 -- X-Spam-Score: -4 () L_EXCH_MF 1, 20 -- X-Scanned-By: CanIt (www . roaringpenguin . com) on 141.209.20.21 ==============================End of - Headers==============================
This is a well known problem that goes back to the early space programs. It has shown-up again since the "world" is switching to no-lead solders due the EU RoHS initiatives. Components cannot contain lead (Pb)in their lead (frame) finishes, so pure tin is the easiest solution. Unfortunately, component manufacturers are not aware of or forgot about the whisker problem also.
One of the most plausible scenarios was presented by Dr. Tu of UCLA at the 2003 ISTFA conference (See "Solder Failure Mechanisms, K.N. Tu, 29th International Symposium for Testing and Failure Analysis - Microelectronics Seminar, ASM International, 2003, P.187). He asserts that the whiskers grow in response to induced stresses in the tin plate deposit, caused by the volumetric changes due to Cu-Sn intermetallic compound formation. A nickel underplate slows this process due to slower Ni-Sn intermetallic compound growth kinetics.
For more info on tin whisker failures, go to http://nepp.nasa.gov/whisker/
Joseph
Joseph M. Oparowski Center for Materials Science Bose Corporation Joseph_Oparowski-at-bose.com The Mountain, M/S 415 Phone: (508) 766-1371 Framingham, MA 01701-9168 Fax: (508) 766-1313
-----Original Message----- X-from: smalinskas-at-yahoo.com [mailto:smalinskas-at-yahoo.com] Sent: Friday, June 16, 2006 10:59 AM To: Oparowski, Joseph
Here are my thoughts:
If this phenomenon can be reliably reproduced, the effect from bias should be easily confirmed or denied.
This appears to be some type of transport mechanism... vapor transport, surface transport, bulk diffusion from the body of the metal, or grain boundary diffusion. It would be interesting to find where tin is being depleted in the solder joint, perhaps using EDS methods and metallographic cross sections. The challenge would be to distinguish tin depletion from normal solder casting segregation.
At the moment I'm assuming the whisker is a single crystal. Perhaps the whisker can be studied using x-ray crystallographic methods - either Laue or TEM methods. This may give a clue to the growth mechanism.
Stu Smalinskas, P.E. Metallurgist SKF Plymouth, Michigan
--- gary-at-gaugler.com wrote:
The whiskers are tin. Silver is added to lead solder to reduce the melting point.
Whiskering occurs at room temperature, high and low humidity and all sorts of conditions. It is a nasty problem for commercial and especially MIL applications. Alternate solders are being studied (different combinations of elements) but their melting point is sufficiently higher than Pb solder to be a real problem.
gary g.
---------------------------------------
At 01:35 PM 6/15/2006, you wrote:
Hi,
I would be curious to see any feedback on this subject, in the name of education.
I have a couple questions. Some of the lead-free solders have silver in them. Are you (or he) sure the whiskers are tin, or could they be silver? When you say "vacuum", are you referring to the low pressures in orbit, or a clean, high vacuum in a chamber of some sort? (or, perhaps, vacuum tube electronic component)
I believe the fields associated with a bias have something to do with it, but don't have any facts, or know what the mechanism is. I wonder if a high (strong) bias, in a vacuum chamber, would accelerate the phenomenon.
Looking forward to learning more. Thanks, Darrell
-----------------------------------------------
beaurega-at-westol.com wrote on 06/15/2006 10:55:59 AM:
Hi, I was asked to look into the morphology and cause of tin whiskers in high tin content lead free solders by a guy I meet. Some of the following questions have been asked for 50 years. Maybe someone out there can share some more recent expertise about tin whiskers.
* What is the root cause of crystalline tin whiskers? (It's probably not resolved yet. There are lots of conflicting opinions and observations on Google.)
* How do you prevent tin whiskers from growing out of almost pure tin solder joints? (He is not that interested in a thin film coating solution.)
* What is the mechanism(s) that can cause 2 micron diameter or less "fibers" (et al) to grow right out of the surface of solder joins to a length of up to one observed to be 10 mm long?
* What are the EDS results of tin whisker analysis?
* Are the EDS analysis results settled science?
* Have solder joints been ion milled to explore the interior of the solder or is this strictly a compression/stress surface phenomena? Or both?
He is NOT interested in high humidity dendritic growth of tin on PCB surfaces reputed to be caused by electric lines and associated equipotential lines. He is interested in the shorts caused by tin whiskers bridging solder joints through the air or a vacuum.
This raises the following big question. * How do you accelerate the growth of tin whiskers so that they can be studied without waiting 1-15 years for them to develop?
Please reply off list with any insights you have on this problem. I don't think this subject will have wide appeal. Thank you in advance for any perspectives that can be shared with him. If you are speculating on a solution or theory, please say so in your reply.
Paul
Disclaimers: I do not work for this guy's company, the guy, or any company or organization working on tin whiskers. I was astonished at the phenomenon, morphology, and how they ruined an obiting satellite. Tin whiskers grow in air or vacuum.
__________________________________________________ Do You Yahoo!? Tired of spam? Yahoo! Mail has the best spam protection around http://mail.yahoo.com
==============================Original Headers============================== 32, 19 -- From smalinskas-at-yahoo.com Fri Jun 16 09:53:13 2006 32, 19 -- Received: from web34410.mail.mud.yahoo.com (web34410.mail.mud.yahoo.com [66.163.178.159]) 32, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5GErDcf012840 32, 19 -- for {microscopy-at-sparc5.microscopy.com} ; Fri, 16 Jun 2006 09:53:13 -0500 32, 19 -- Received: (qmail 66812 invoked by uid 60001); 16 Jun 2006 14:53:12 -0000 32, 19 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 32, 19 -- s=s1024; d=yahoo.com; 32, 19 -- h=Message-ID:Received:Date:From:Subject:To:In-Reply-To:MIME-Version:Cont ent-Type:Content-Transfer-Encoding; 32, 19 -- b=ZaqnxzqR/WQm9UGzGSDQ9QDR2u7EqRGDp1gUTTsmNfHkKxWlQH1BwTvFTXVYpBXKX4SmcK wCyVc8BUbFNLgWPbdC7Ww1zCaT+83IV0/q39xwxFYWr3jIz33E5TV99VdjgCAJSXnlo9AnVL NU6uYPnQKuaZuNjlylNT6ruPENBMQ= ; 32, 19 -- Message-ID: {20060616145312.66810.qmail-at-web34410.mail.mud.yahoo.com} 32, 19 -- Received: from [141.151.33.213] by web34410.mail.mud.yahoo.com via HTTP; Fri, 16 Jun 2006 07:53:12 PDT 32, 19 -- Date: Fri, 16 Jun 2006 07:53:12 -0700 (PDT) 32, 19 -- From: Kestutis Smalinskas {smalinskas-at-yahoo.com} 32, 19 -- Subject: Re: [Microscopy] Tin Whiskers in high tin content solders. 32, 19 -- To: microscopy-at-ns.microscopy.com 32, 19 -- In-Reply-To: {200606152112.k5FLCRUK023781-at-ns.microscopy.com} 32, 19 -- MIME-Version: 1.0 32, 19 -- Content-Type: text/plain; charset=iso-8859-1 32, 19 -- Content-Transfer-Encoding: 8bit ==============================End of - Headers==============================
==============================Original Headers============================== 46, 23 -- From Joseph_Oparowski-at-bose.com Fri Jun 16 13:33:00 2006 46, 23 -- Received: from BOSEMX02.bose.com (bosemx02.bose.com [139.68.146.21]) 46, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with SMTP id k5GIWx1W019696 46, 23 -- for {Microscopy-at-Microscopy.Com} ; Fri, 16 Jun 2006 13:33:00 -0500 46, 23 -- Received: from USMAFREXMB02.bose.com ([139.68.132.238]) by USMAFREXCN02.bose.com with Microsoft SMTPSVC(6.0.3790.211); 46, 23 -- Fri, 16 Jun 2006 14:32:58 -0400 46, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 46, 23 -- Content-class: urn:content-classes:message 46, 23 -- MIME-Version: 1.0 46, 23 -- Content-Type: text/plain; 46, 23 -- charset="us-ascii" 46, 23 -- Subject: RE: [Microscopy] Re: Tin Whiskers in high tin content solders. 46, 23 -- Date: Fri, 16 Jun 2006 14:32:58 -0400 46, 23 -- Message-ID: {210C73AB4F6E0B4FBCB77B969C0BD84A029A7E2D-at-USMAFREXMB02.bose.com} 46, 23 -- X-MS-Has-Attach: 46, 23 -- X-MS-TNEF-Correlator: 46, 23 -- Thread-Topic: [Microscopy] Re: Tin Whiskers in high tin content solders. 46, 23 -- Thread-Index: AcaRVYq5+zcAKxcmTGi61vt0DqKzBAAGnAEA 46, 23 -- From: "Oparowski, Joseph" {Joseph_Oparowski-at-bose.com} 46, 23 -- To: {Microscopy-at-Microscopy.Com} 46, 23 -- X-OriginalArrivalTime: 16 Jun 2006 18:32:58.0282 (UTC) FILETIME=[4A70F0A0:01C69173] 46, 23 -- Content-Transfer-Encoding: 8bit 46, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5GIWx1W019696 ==============================End of - Headers==============================
We are starting a project looking at otolith microstructure, and I was wondering if folks have recommendations for mounting resins. We have one reference that recommends spurr's resin. Does anybody have experience with this type of preparation or a recommendation?
Karl Hagglund Biological Sciences, SC-102B Northern Kentucky University, Nunn Drive Highland Heights, KY 41099 859-572-5238 http://semlab.nku.edu
==============================Original Headers============================== 4, 23 -- From hagglundk1-at-nku.edu Fri Jun 16 15:25:46 2006 4, 23 -- Received: from mailfe2.nku.edu (mailfe2.nku.edu [192.122.237.68]) 4, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5GKPkgG005767 4, 23 -- for {microscopy-at-microscopy.com} ; Fri, 16 Jun 2006 15:25:46 -0500 4, 23 -- Received: from mailfac1.nku.edu ([172.28.102.15]) by mailfe2.nku.edu with Microsoft SMTPSVC(6.0.3790.1830); 4, 23 -- Fri, 16 Jun 2006 16:25:45 -0400 4, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 4, 23 -- Content-class: urn:content-classes:message 4, 23 -- MIME-Version: 1.0 4, 23 -- Content-Type: text/plain; 4, 23 -- charset="US-ASCII" 4, 23 -- Subject: LM/SEM Mounting Otoliths 4, 23 -- Date: Fri, 16 Jun 2006 16:25:45 -0400 4, 23 -- Message-ID: {F01E4499C4EC5842A8AB7198141AC72F35875F-at-mailfac1.hh.nku.edu} 4, 23 -- X-MS-Has-Attach: 4, 23 -- X-MS-TNEF-Correlator: 4, 23 -- Thread-Topic: LM/SEM Mounting Otoliths 4, 23 -- Thread-Index: AcaRgwv65/QFS15YQtiOCGTvbfq4SQ== 4, 23 -- From: "Karl Hagglund" {hagglundk1-at-nku.edu} 4, 23 -- To: {microscopy-at-microscopy.com} 4, 23 -- X-OriginalArrivalTime: 16 Jun 2006 20:25:46.0084 (UTC) FILETIME=[0C5D9240:01C69183] 4, 23 -- Content-Transfer-Encoding: 8bit 4, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5GKPkgG005767 ==============================End of - Headers==============================
It's not too late to enter the International Metallographic Contest and Exhibit co-sponsored since 1972 by the International Metallographic Society and ASM International. The contest will be held in conjunction with the 39th annual technical meeting of the IMS and the M&M 2006 meeting this summer in Chicago. Twelve categories of competition. Best in show receives $3000 and the prestigious Jaquet-Lucas Award. Cash awards for first, second, and third place winners in eleven of the categories. Entries are prominently displayed during the M&M 2006 meeting and again in the fall during the ASM Annual Event. Deadline for entries is July 17. For additional information including rules, tips for creating a winning entry, judging guidelines, and examples of winning entries contact me or visit http://www.internationalmetallographicsociety.org/contest.html.
Jeff Stewart International Metallographic Contest Chair Metallographic Laboratory Manager Stern-Leach Co. 49 Pearl Street Attleboro, MA 02703 USA Phone: 508-222-7400 extension 1329 FAX: 508-699-4030
==============================Original Headers============================== 4, 21 -- From jeff-at-metallography.com Sat Jun 17 06:38:18 2006 4, 21 -- Received: from vw-cust-mail-app2-plano-126.emailscience.com (vw-cust-mail-app2-plano-126.emailscience.com [207.235.126.31]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5HBcI74012896 4, 21 -- for {Microscopy-at-microscopy.com} ; Sat, 17 Jun 2006 06:38:18 -0500 4, 21 -- Received: (qmail 95723 invoked by uid 1009); 17 Jun 2006 11:26:30 -0000 4, 21 -- Received: from unknown (HELO jstewart) (4.154.215.5) 4, 21 -- by vw-cust-mail-app2-plano.emailscience.com with SMTP; 17 Jun 2006 11:26:30 -0000 4, 21 -- Message-ID: {002001c69202$c801f840$959610ac-at-sternleach.com} 4, 21 -- Reply-To: "Jeff Stewart" {jeff-at-metallography.com} 4, 21 -- From: "Jeff Stewart" {jeff-at-metallography.com} 4, 21 -- To: {Microscopy-at-microscopy.com} 4, 21 -- Subject: International Metallographic Contest announcement 4, 21 -- Date: Sat, 17 Jun 2006 07:39:33 -0400 4, 21 -- MIME-Version: 1.0 4, 21 -- Content-Type: text/plain; 4, 21 -- charset="iso-8859-1" 4, 21 -- Content-Transfer-Encoding: 7bit 4, 21 -- X-Priority: 3 4, 21 -- X-MSMail-Priority: Normal 4, 21 -- X-Mailer: Microsoft Outlook Express 6.00.2800.1409 4, 21 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2800.1409 ==============================End of - Headers==============================
Our Hitachi S-4700 is suffering from 60 Hz EMF's in our lab. We see jagged lines in the horizontal direction at mags } /= to 150kX. I just had a vibration survey done and the results are within Hitachi specifications.
Do any of you pay particular attention to movement (of people, people in chairs, talking, etc.) when you are operating the SEM } 200kX? We pay particular attention to this in our HR-TEM lab. Just wondering if it is an issue in the FESEM lab too.
I'm looking for solutions. Cheap(er) DYI solutions. Any ideas?
Thanks!
OWen
Owen P. Mills Director, Materials Characterization & Fabrication Facilities Electron Optics Engineer, Applied Chemical & Morphological Analysis Laboratory
Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
==============================Original Headers============================== 12, 31 -- From opmills-at-mtu.edu Mon Jun 19 10:34:23 2006 12, 31 -- Received: from node8.mtu.edu (node8.mtu.edu [141.219.69.8]) 12, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JFYNfX014474 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 10:34:23 -0500 12, 31 -- Received: from node5.edge.dcsint.mtu.edu (node5.mtu.edu [141.219.69.5]) 12, 31 -- by node8.mtu.edu (8.13.1/8.13.1) with ESMTP id k5JFYNe6005289 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:23 -0400 12, 31 -- Received: from node34.edge.dcsint.mtu.edu (node34.mtu.edu [141.219.69.34]) 12, 31 -- by node5.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.8) with ESMTP id k5JFYMjZ018584 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:22 -0400 12, 31 -- Received: from mail.mtu.edu (campus4.mtu.edu [141.219.70.4]) 12, 31 -- by node34.edge.dcsint.mtu.edu (8.12.11/8.12.11) with ESMTP id k5JFYLtJ011265 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:22 -0400 12, 31 -- (envelope-from opmills-at-mtu.edu) 12, 31 -- Received: from node6.edge.dcsint.mtu.edu (node6.mtu.edu [141.219.69.6]) 12, 31 -- by mail.mtu.edu (8.13.6/8.11.6) with ESMTP id k5JFYLlX008693 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:21 -0400 (EDT) 12, 31 -- Received: from [141.219.192.45] (opmills.eecn.sabo.mtu.edu [141.219.192.45]) 12, 31 -- by node6.edge.dcsint.mtu.edu (8.12.11.20060308/8.12.3/auth_ssl.mc v1.0) with ESMTP id k5JFYLJ6022941 12, 31 -- for {Microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 11:34:21 -0400 12, 31 -- Mime-Version: 1.0 (Apple Message framework v750) 12, 31 -- Content-Transfer-Encoding: 7bit 12, 31 -- Message-Id: {F22556D6-9A4C-4046-94CF-A6BF69E16726-at-mtu.edu} 12, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 31 -- To: Microscopy Listserver {Microscopy-at-microscopy.com} 12, 31 -- From: "Owen P. Mills" {opmills-at-mtu.edu} 12, 31 -- Subject: SEM - electromagnetic fields 12, 31 -- Date: Mon, 19 Jun 2006 11:34:19 -0400 12, 31 -- X-Mailer: Apple Mail (2.750) 12, 31 -- X-PMX-Version: 4.7.1.128075, Antispam-Engine: 2.4.0.264935, Antispam-Data: 2006.6.19.81433 12, 31 -- X-PerlMx-Spam: Gauge=IIIIIII, Probability=7%, Report='__C230066_P5 0, __CP_URI_IN_BODY 0, __CT 0, __CTE 0, __CT_TEXT_PLAIN 0, __HAS_MSGID 0, __HAS_X_MAILER 0, __MIME_TEXT_ONLY 0, __MIME_VERSION 0, __SANE_MSGID 0' ==============================End of - Headers==============================
I am sorry to say but round the world one of the problems that we all have is electromagnetic fields. To prove your problem compare working at a long WD with working at a very short WD. If the problem changes for the better at short WD it is a field problem. If it stays the same its most probably vibration or an instrument fault.
Check laboratories above, below and on all sides to track down the equipment that may be causing the problem, try switching equipment off.
As a warning to all, magnetic fields now prove to be the biggest FEG installation problem so always have a professional check your site. Remember the field will change by the inverse square so in some cases to move the instrument in the room may help. Field reduction systems work if a field is not over strong but the best route is to find a room which is truly field free.
Good luck
Steve Chapman Senior Consultant Protrain For electron microscopy consultancy and training world wide Tel +44 1280 816512 Fax +44 1280 814007 Mobile +44 7711 606967 Web www.emcourses.com
----- Original Message ----- X-from: {opmills-at-mtu.edu} To: {protrain-at-emcourses.com} Sent: Monday, June 19, 2006 4:37 PM
Hi All, I have been trying unsuccessfully to immuno-gold label HUVEC cells that have been embedded in LR White resin. The cells were plated and grown to supposed confluence, incubated with primary antibodies to PECAM and another adherence molecule, then fixed in 2% pfa, 0.5% glut., dehydrated, & embedded. The resin was cured at 50 deg. C overnight. En face sections were hydrated & blocked with buffer, then incubated with gold-tagged secondary Ab (anti-mouse or anti-rabbit). The sections were contrasted with aqueous Ur. Ac. then Pb citrate. I"ve seen nothing. For secondaries, I tried both F(ab)2s and whole IgG. I have been very successful with the same F(ab)2 secondaries on other samples processed the same way, but on which both primary and secondary Ab incubations were done on grid. In the past, we have successfully localized each of these primary Abs with peroxidase-DAB labelling prior to stringent fixation and embedding in Spurr's resin.
One of the problems is that HUVECs (human umbilical cord endothelial cells) are so darned flat...(less than 0.3 micrometers except for the nucleus) that its easy to cut right through them just trying to get a smooth section with no holes off of the raw block face. Another problem is that the proteins of interest are located at the cell junctions, in little vesicles and its not always easy to find cell-cell contacts.
My question (finally) is this: Has anyone tried to do an immuno-gold approach after immuno-peroxidase? We thought we'd like to try repeating the successful labelling of PECAM with DAB, pre-embedding, then try for the other Ab post-embedding.
Any ideas? (I'm sure there are....) thanks in advance, Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy & Histology Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175 http://www.cornellcelldevbiology.org http://www.cornellbiochem.org
==============================Original Headers============================== 4, 21 -- From lcgould-at-med.cornell.edu Mon Jun 19 13:48:43 2006 4, 21 -- Received: from smtp-gw1.med.cornell.edu (smtp-gw1.med.cornell.edu [140.251.3.74]) 4, 21 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JImg1C014989 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 13:48:43 -0500 4, 21 -- Received: from mpx1.med.cornell.edu (biglb-vlan511vip.med.cornell.edu [140.251.11.120]) 4, 21 -- by smtp-gw1.med.cornell.edu (Switch-3.1.8/Switch-3.1.7) with ESMTP id k5JImWJi025260 4, 21 -- for {microscopy-at-microscopy.com} ; Mon, 19 Jun 2006 14:48:36 -0400 (EDT) 4, 21 -- Received: from [140.251.48.23] (mac110773.med.cornell.edu [140.251.48.23]) 4, 21 -- by mpx1.med.cornell.edu 4, 21 -- (Sun Java System Messaging Server 6.1 HotFix 0.11 (built Jan 28 2005)) 4, 21 -- with ESMTPA id {0J1400HVGEWW1R30-at-mpx1.med.cornell.edu} for 4, 21 -- microscopy-at-microscopy.com; Mon, 19 Jun 2006 14:48:32 -0400 (EDT) 4, 21 -- Date: Mon, 19 Jun 2006 14:41:57 -0400 4, 21 -- From: Leona Cohen-Gould {lcgould-at-med.cornell.edu} 4, 21 -- Subject: EM: immuno labelling DAB & gold 4, 21 -- Sender: lcgould-at-med.cornell.edu 4, 21 -- To: Microscopy Listserver {microscopy-at-microscopy.com} 4, 21 -- Message-id: {p0623091ac0bc9c974d8d-at-[140.251.48.23]} 4, 21 -- MIME-version: 1.0 4, 21 -- Content-type: text/plain; charset=us-ascii; format=flowed 4, 21 -- X-PMX-Version: 5.1.2.240295, Antispam-Engine: 2.3.0.1, Antispam-Data: 2006.6.19.113432 ==============================End of - Headers==============================
1) I would worry that the DAB black mass would obscure the gold. I have not tried it, but it seems like this would be a problem. 2) Why not do primary and secondary Ab before fix, then fix hard and go into Epon? Your gold would be in the Epon. No post-cutting Ab work. 3) I would recommend cutting cross-sections of the cells. Then you do not have to worry about missing the flat cells when facing the block. Also, cell-cell junctions are much easier to see. I have protocols to do this if you want them.
David
_____________________
David Elliott Ph.D. Assistant Professor Department of Cell Biology and Anatomy University of Arizona College of Medicine PO Box 245004 Tucson, AZ 85724
Voice: 520-626-7870 Fax: 520-626-2097
On Jun 19, 2006, at 11:54 AM, lcgould-at-med.cornell.edu wrote:
} } Hi All, } I have been trying unsuccessfully to immuno-gold label HUVEC cells } that have been embedded in LR White resin. The cells were plated and } grown to supposed confluence, incubated with primary antibodies to } PECAM and another adherence molecule, then fixed in 2% pfa, 0.5% } glut., dehydrated, & embedded. The resin was cured at 50 deg. C } overnight. En face sections were hydrated & blocked with buffer, } then incubated with gold-tagged secondary Ab (anti-mouse or } anti-rabbit). The sections were contrasted with aqueous Ur. Ac. then } Pb citrate. I"ve seen nothing. } For secondaries, I tried both F(ab)2s and whole IgG. I have been } very successful with the same F(ab)2 secondaries on other samples } processed the same way, but on which both primary and secondary Ab } incubations were done on grid. } In the past, we have successfully localized each of these primary Abs } with peroxidase-DAB labelling prior to stringent fixation and } embedding in Spurr's resin. } } One of the problems is that HUVECs (human umbilical cord endothelial } cells) are so darned flat...(less than 0.3 micrometers except for the } nucleus) that its easy to cut right through them just trying to get a } smooth section with no holes off of the raw block face. Another } problem is that the proteins of interest are located at the cell } junctions, in little vesicles and its not always easy to find } cell-cell contacts. } } My question (finally) is this: Has anyone tried to do an immuno-gold } approach after immuno-peroxidase? We thought we'd like to try } repeating the successful labelling of PECAM with DAB, pre-embedding, } then try for the other Ab post-embedding. } } Any ideas? (I'm sure there are....) } thanks in advance, } Lee } -- } Leona Cohen-Gould, M.S. } Sr. Staff Associate } Director, Electron Microscopy & Histology Core Facility } Manager, Optical Microscopy Core Facility } Joan & Sanford I. Weill Medical College } of Cornell University } voice (212)746-6146 } fax (212)746-8175 } http://www.cornellcelldevbiology.org } http://www.cornellbiochem.org
==============================Original Headers============================== 12, 22 -- From Elliott-at-arizona.edu Mon Jun 19 14:25:53 2006 12, 22 -- Received: from smtpgate.email.arizona.edu (deagol.email.Arizona.EDU [128.196.133.142]) 12, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JJPrMx001845 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 14:25:53 -0500 12, 22 -- Received: from localhost (legolas.email.arizona.edu [10.0.0.224]) 12, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 9E5D5E4B48E 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 12:25:52 -0700 (MST) 12, 22 -- Received: from [150.135.145.126] (unknown [150.135.145.126]) 12, 22 -- by smtpgate.email.arizona.edu (Postfix) with ESMTP id 5748BE4B45B 12, 22 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 12:25:51 -0700 (MST) 12, 22 -- Mime-Version: 1.0 (Apple Message framework v750) 12, 22 -- In-Reply-To: {200606191854.k5JIsHFb022009-at-ns.microscopy.com} 12, 22 -- References: {200606191854.k5JIsHFb022009-at-ns.microscopy.com} 12, 22 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 12, 22 -- Message-Id: {6807EC57-D3D2-43A6-A1E0-8EEE3EED6C93-at-arizona.edu} 12, 22 -- Content-Transfer-Encoding: 7bit 12, 22 -- From: David Elliott {Elliott-at-arizona.edu} 12, 22 -- Subject: Re: [Microscopy] EM: immuno labelling DAB & gold 12, 22 -- Date: Mon, 19 Jun 2006 12:25:50 -0700 12, 22 -- To: Microscopy ListServer {Microscopy-at-MSA.Microscopy.Com} 12, 22 -- X-Mailer: Apple Mail (2.750) 12, 22 -- X-Virus-Scanned: amavisd-new at email.arizona.edu ==============================End of - Headers==============================
A friend wants to upgrade/keep going their Zeiss DSM 940A SEM.
First, they would like to know of any recommendations for a digital image acquisition upgrade available for this model. Contact off-list by commercial companies welcome.
Also, they are interested in anyone they can contact who might have the technical service manual, electrical schematics, etc., for this machine and/or spare parts that they might purchase or have donated. They are in Maine.
Mahalo! Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
==============================Original Headers============================== 8, 19 -- From tina-at-pbrc.hawaii.edu Mon Jun 19 16:35:13 2006 8, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JLZBUg023698 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 16:35:12 -0500 8, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) 8, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k5JLZ6HC009676 8, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 11:35:06 -1000 (HST) 8, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k5JLZ5iU009673 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 11:35:05 -1000 (HST) 8, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs 8, 19 -- Date: Mon, 19 Jun 2006 11:35:04 -1000 (HST) 8, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} 8, 19 -- X-Sender: tina-at-halia 8, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} 8, 19 -- Subject: Digital acquisition/parts for Zeiss DSM 940A 8, 19 -- Message-ID: {Pine.GSO.4.21.0606191129200.9568-100000-at-halia} 8, 19 -- MIME-Version: 1.0 8, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
My, isn't this a popular instrument to keep up and running... I have asked the same in the past few months...
Tell your friend in Maine to contact me, as I have been actively looking for replacement parts for this instrument, I have run across a fair amount of information. In fact, there is a group in Maine that has a Zeiss DM940A that they may be interested in getting to know....
Also, I would be interested in any information you may get from your request for digital image acquisition.
dj
On Mon, 19 Jun 2006, tina-at-pbrc.hawaii.edu wrote:
} } } } ---------------------------------------------------------------------------- } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------------- } } Hi, All- } } A friend wants to upgrade/keep going their Zeiss DSM 940A SEM. } } First, they would like to know of any recommendations for a digital image } acquisition upgrade available for this model. Contact off-list by } commercial companies welcome. } } Also, they are interested in anyone they can contact who might have the } technical service manual, electrical schematics, etc., for this machine } and/or spare parts that they might purchase or have donated. They are in } Maine. } } Mahalo! } Tina } } **************************************************************************** } * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * } * Biological Electron Microscope Facility * (808) 956-6251 * } * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* } **************************************************************************** } } } } ==============================Original Headers============================== } 8, 19 -- From tina-at-pbrc.hawaii.edu Mon Jun 19 16:35:13 2006 } 8, 19 -- Received: from halia.pbrc.hawaii.edu (halia.pbrc.hawaii.edu [128.171.22.7]) } 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5JLZBUg023698 } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 16:35:12 -0500 } 8, 19 -- Received: from halia.pbrc.hawaii.edu (localhost [127.0.0.1]) } 8, 19 -- by halia.pbrc.hawaii.edu (8.12.11/8.12.11) with ESMTP id k5JLZ6HC009676 } 8, 19 -- (version=TLSv1/SSLv3 cipher=DHE-RSA-AES256-SHA bits=256 verify=NO) } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 11:35:06 -1000 (HST) } 8, 19 -- Received: from localhost by halia.pbrc.hawaii.edu (8.12.11/8.12.11/Submit) with ESMTP id k5JLZ5iU009673 } 8, 19 -- for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 19 Jun 2006 11:35:05 -1000 (HST) } 8, 19 -- X-Authentication-Warning: halia.pbrc.hawaii.edu: tina owned process doing -bs } 8, 19 -- Date: Mon, 19 Jun 2006 11:35:04 -1000 (HST) } 8, 19 -- From: Tina Carvalho {tina-at-pbrc.hawaii.edu} } 8, 19 -- X-Sender: tina-at-halia } 8, 19 -- To: Microscopy Listserver {Microscopy-at-MSA.Microscopy.Com} } 8, 19 -- Subject: Digital acquisition/parts for Zeiss DSM 940A } 8, 19 -- Message-ID: {Pine.GSO.4.21.0606191129200.9568-100000-at-halia} } 8, 19 -- MIME-Version: 1.0 } 8, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII } ==============================End of - Headers============================== }
==============================Original Headers============================== 8, 19 -- From dljones-at-bestweb.net Tue Jun 20 07:33:03 2006 8, 19 -- Received: from vms046pub.verizon.net (vms046pub.verizon.net [206.46.252.46]) 8, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5KCX3gS021164 8, 19 -- for {Microscopy-at-microscopy.com} ; Tue, 20 Jun 2006 07:33:03 -0500 8, 19 -- Received: from localhost ([71.249.29.149]) 8, 19 -- by vms046.mailsrvcs.net (Sun Java System Messaging Server 6.2-4.02 (built Sep 8, 19 -- 9 2005)) with ESMTPA id {0J1500ES8S6LJ2CA-at-vms046.mailsrvcs.net} for 8, 19 -- Microscopy-at-microscopy.com; Tue, 20 Jun 2006 07:32:59 -0500 (CDT) 8, 19 -- Date: Tue, 20 Jun 2006 08:43:52 -0400 (Eastern Standard Time) 8, 19 -- From: "David L. Jones" {dljones-at-bestweb.net} 8, 19 -- Subject: Re: [Microscopy] Digital acquisition/parts for Zeiss DSM 940A 8, 19 -- In-reply-to: {200606192141.k5JLfBEa030525-at-ns.microscopy.com} 8, 19 -- X-X-Sender: dljones-at-pop-croton.bestweb.net 8, 19 -- To: tina-at-pbrc.hawaii.edu 8, 19 -- Cc: Microscopy-at-microscopy.com 8, 19 -- Message-id: {Pine.WNT.4.64.0606200837420.3168-at-H-F1} 8, 19 -- MIME-version: 1.0 8, 19 -- Content-type: TEXT/PLAIN; charset=US-ASCII; format=flowed 8, 19 -- References: {200606192141.k5JLfBEa030525-at-ns.microscopy.com} ==============================End of - Headers==============================
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cloverbags-at-yahoo.com) from http://www.microscopy.com/Ask-A-Microscopist/Ask-A-Microscopist.html on Tuesday, June 20, 2006 at 09:34:21 ---------------------------------------------------------------------------
Email: cloverbags-at-yahoo.com Name: raffaela
Organization: cebu doctors' university
Education: Undergraduate College
Location: Cebu City, Philippines,Asia
Question: what is the pH of a cedarwood oilJuniperus virginiana) for oil immersion objective? what is the pH of lemongrass oil(Cymbopogon citratus)?
Dear friends, please find the following CALL for PAPERS for Journal of Biomedical Optics All the best. Alby and Peri
---- May/June 2007
Visible Fluorescent Proteins
Guest Editors:
Alberto "Alby" Diaspro University of Genoa Department of Physics MicroScoBio-LAMBS/IFOM-NANOMED Via Dodecaneso 33 16146 Genova, Italy Tel: +39 10 353 6426 Fax: +39 10 314 218 E-mail: diaspro-at-fisica.unige.it
Ammasi Periasamy University of Virginia Keck Center for Cellular Imaging Biology Department Gilmer Hall (064), McCormick Rd Charlottesville, VA 22904 Tel: 434-243-7602 Fax: 434-982-5210 E-mail: ap3t-at-virginia.edu
Call for Papers: This special section will focus on the utilization of visible fluorescent proteins (VFPs) in fluorescence microscopy and spectroscopy. The use of fluorescent proteins and live-cell imaging has greatly improved our understanding of cell functioning in recent years. Discoveries in different organisms and development of fluorescent proteins are revolutionizing the study of cell behavior by providing convenient in vivo markers for gene expression and protein targeting in intact cells and organisms. VFPs coupled to three-dimensional fluorescence imaging methods such as confocal and multiphoton excitation microscopy are very effective in live-cell imaging. Moreover, the need to understand the fluorescence mechanism under different conditions has inspired a renaissance in single- molecule methods. Areas of interest also include papers related to fluorescence resonance energy transfer (FRET) and fluorescence recovery after photobleaching (FRAP) applications, VFP photophysical characterization, and photoconvertible and photactivatable mutants. In vivo applications are welcome as well as comparisons with potential competitors such as new fluorescent molecules and quantum dots.
Manuscripts due September 1, 2006
---------
--------------------------------------------- [...] Dance with wolves and count the stars, including the unseen...(L.Ferlinghetti, Challenges to young poet, 2001) --------------------------------------------- Alberto Diaspro, MicroScoBIO Research Center, LAMBS-IFOM, Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genova, Italy facsimile +39-010314218 - voice +39-0103536426/480/309; URL: http://www.lambs.it http://www.wiley.com/WileyCDA/WileyTitle/productCd-0471409200.html ----------------------------------------------
==============================Original Headers============================== 15, 31 -- From diaspro-at-fisica.unige.it Wed Jun 21 18:27:29 2006 15, 31 -- Received: from phobos.ge.infm.it (phobos.ge.infm.it [193.205.153.2]) 15, 31 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5LNRSGJ009011 15, 31 -- for {microscopy-at-microscopy.com} ; Wed, 21 Jun 2006 18:27:28 -0500 15, 31 -- Received: from [193.205.153.216] (albypc.ge.infm.it [193.205.153.216]) 15, 31 -- by phobos.ge.infm.it (8.11.6/8.11.0) with ESMTP id k5L6ZXh10248; 15, 31 -- Wed, 21 Jun 2006 08:35:33 +0200 15, 31 -- Mime-Version: 1.0 (Apple Message framework v750) 15, 31 -- Content-Transfer-Encoding: 7bit 15, 31 -- Message-Id: {85421244-1096-48EB-89F8-934BB7B5F916-at-fisica.unige.it} 15, 31 -- Content-Type: text/plain; charset=US-ASCII; delsp=yes; format=flowed 15, 31 -- To: mplsm-users-at-yahoogroups.com, 15, 31 -- Confocal List Microscopy {CONFOCAL-at-LISTSERV.BUFFALO.EDU} , 15, 31 -- microscopy-at-microscopy.com, Ronzitti Emiliano {ronzi81-at-virgilio.it} , 15, 31 -- Cella Francesca {fra_cella-at-yahoo.it} , 15, 31 -- Mondal Partha Pratim {partha-at-fisica.unige.it} , 15, 31 -- Magrassi Raffaella {magrassi-at-fisica.unige.it} , 15, 31 -- Caorsi Valentina {caorsi-at-fisica.unige.it} , 15, 31 -- Diaspro Alberto {diaspro-at-fisica.unige.it} , 15, 31 -- Bianchini Paolo {bianchini-at-ge.infm.it} , 15, 31 -- Gattodub Gattodub {gattodub-at-email.it} , 15, 31 -- Mazza Davide {mazza-at-fisica.unige.it} , 15, 31 -- Morotti Federica {morottina-at-yahoo.it} , 15, 31 -- Testa Ilaria {ilaria.testa-at-email.it} , 15, 31 -- Vicidomini Giuseppe {vicidomini-at-fisica.unige.it} , 15, 31 -- Testa Ilaria {testa-at-fisica.unige.it} , 15, 31 -- Mario Faretta {mario.faretta-at-ifom-ieo-campus.it} 15, 31 -- From: diaspro {diaspro-at-fisica.unige.it} 15, 31 -- Subject: JBO special issue - call for papers 15, 31 -- Date: Wed, 21 Jun 2006 08:30:47 +0200 15, 31 -- X-Mailer: Apple Mail (2.750) ==============================End of - Headers==============================
A enthusiastic construction worker cut a major telecom trunk line out here in the suburbs of Chicago a little over 24 hours ago. Took out ALL telecom links, phone lines, networks the whole system. It made for a very quiet and annoying 24 hours here. Amazing how rapidly wedded we become to communications links.
Anyway, although the Listserver was running it was not able to "talk to anyone".
If you attempted posting anything during the las 24 hours there is a good likelihood it was held in queue somewhere on the net and it may get resent. If your not sure then feel free to repost any messages.
Cheers...
Nestor Your Friendly Neighborhood SysOp (who had a very quiet 24 + hours )
==============================Original Headers============================== 9, 11 -- From zaluzec-at-microscopy.com Wed Jun 21 19:57:21 2006 9, 11 -- Received: from [206.69.208.22] (mac22.zaluzec.com [206.69.208.22]) 9, 11 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5M0vJXJ023886 9, 11 -- for {microscopy-at-microscopy.com} ; Wed, 21 Jun 2006 19:57:20 -0500 9, 11 -- Mime-Version: 1.0 9, 11 -- Message-Id: {p06110402c0bf988fd43d-at-[206.69.208.22]} 9, 11 -- Date: Wed, 21 Jun 2006 19:57:19 -0500 9, 11 -- To: microscopy-at-microscopy.com 9, 11 -- From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com} 9, 11 -- Subject: Administrivia: Network Connections have been restored 9, 11 -- Content-Type: text/plain; charset="us-ascii" ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://microscopy.com/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both kjl226-at-vt.edu as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: kjl226-at-vt.edu Name: Kathy
Organization: Virginia Tech
Title-Subject: [Filtered] Salary ranges
Question: What is a fair salary for a supervisory position, where the person will run an Electron Microscope Service Laboratory? This lab will offer all phases of processing of SEM amd TEM samples. Sectioning of the TEM samples, thick and/or thins, negative staining. Running both scopes and assist with use of the scopes including capturing digital images.
This person will also be responible for ordering all supplies, undating SOP's as needed, keeping labortories maintained, helping students with projects, keeping inventory of equipment, keeping all equipment in good working condition, calling service personnel when scopes on service contracts need work, and calulating bills for EM services.
Check out the Microscopy Today archives for the salary survey done in 2004 by Ron Anderson and Barb Foster. It contains a lot of valuable comparison data that should help you determine an appropriate salary range for your geographical location and responsibilities.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907 http://www.agriculture.purdue.edu/microscopy
On 6/21/06 9:04 PM, "kjl226-at-vt.edu" {kjl226-at-vt.edu} wrote:
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==============================Original Headers============================== 7, 22 -- From dsherman-at-purdue.edu Wed Jun 21 20:46:36 2006 7, 22 -- Received: from 1061exfe01.adpc.purdue.edu (1061exfe01.adpc.purdue.edu [128.210.63.221]) 7, 22 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5M1kaBE012448 7, 22 -- for {microscopy-at-microscopy.com} ; Wed, 21 Jun 2006 20:46:36 -0500 7, 22 -- Received: from exchange.purdue.edu ([172.21.6.24]) by 1061exfe01.adpc.purdue.edu with Microsoft SMTPSVC(6.0.3790.211); 7, 22 -- Wed, 21 Jun 2006 21:46:35 -0400 7, 22 -- Received: from 74.132.211.81 ([74.132.211.81]) by EXCH04.purdue.lcl ([172.21.6.26]) via Exchange Front-End Server exch.itap.purdue.edu ([128.210.63.104]) with Microsoft Exchange Server HTTP-DAV ; 7, 22 -- Thu, 22 Jun 2006 01:46:34 +0000 7, 22 -- User-Agent: Microsoft-Entourage/11.2.3.060209 7, 22 -- Date: Wed, 21 Jun 2006 21:46:37 -0400 7, 22 -- Subject: Re: [Microscopy] viaWWW: Salary ranges 7, 22 -- From: Debby Sherman {dsherman-at-purdue.edu} 7, 22 -- To: {kjl226-at-vt.edu} , "message to: MSA list" {microscopy-at-microscopy.com} 7, 22 -- Message-ID: {C0BF6EBD.37F0%dsherman-at-purdue.edu} 7, 22 -- Thread-Topic: [Microscopy] viaWWW: Salary ranges 7, 22 -- Thread-Index: AcaVnbLe8ZosfgGQEduvUQAKlcoUxg== 7, 22 -- In-Reply-To: {200606220104.k5M14nlM005892-at-ns.microscopy.com} 7, 22 -- Mime-version: 1.0 7, 22 -- Content-type: text/plain; 7, 22 -- charset="US-ASCII" 7, 22 -- Content-transfer-encoding: 7bit 7, 22 -- X-OriginalArrivalTime: 22 Jun 2006 01:46:35.0710 (UTC) FILETIME=[B21A25E0:01C6959D] ==============================End of - Headers==============================
Question: what is the pH of a cedarwood oilJuniperus virginiana) for oil immersion objective? what is the pH of lemongrass oil(Cymbopogon citratus)?
1. There are three types of cedar oil:
Virginia cedarwood oil (Juniperus virginiana), Texas cedarwood oil (Juniperus ashei or mexicana), and Western red cedar (Thuja plicata)
I have cedar oil (Texas) from Polysciences. Their MSDS does not list a pH. Nor does Merck list one. The Sigma Aldrich MSDS does not list one, nor does the Libety Nautral Products (Virginia), nor does JT Baker, nor does Well, Naturally Products (Virginia & Texas & Western).
I will try to get a pH on my Texas cedar oil next week and report this at least.
The constituents are mainly cedrene (a terpene) and cedral (cedar camphor).
2. regarding lemongrass oil:
I have found no pH. Well, Naturally Products does not have it in their MSDS.
3. I have a client that extracts, sells, distributes these oils and will check for their data as well next week.
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
This message is from pH2. This message and any attachments may contain legally privileged or confidential information, and are intended only for the individual or entity identified above as the addressee. If you are not the addressee, or if this message has been addressed to you in error, you are not authorized to read, copy, or distribute this message and any attachments, and we ask that you please delete this message and attachments (including all copies) and notify the sender by return e-mail or by phone at 317-752-6386. Delivery of this message and any attachments to any person other than the intended recipient(s) is not intended in any way to waive confidentiality or a privilege. All personal messages express views only of the sender, which are not to be attributed to pH2 and may not be copied or distributed without this statement.
==============================Original Headers============================== 21, 32 -- From ph2-at-sprynet.com Wed Jun 21 22:34:37 2006 21, 32 -- Received: from elasmtp-junco.atl.sa.earthlink.net (elasmtp-junco.atl.sa.earthlink.net [209.86.89.63]) 21, 32 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5M3Yano024776 21, 32 -- for {Microscopy-at-Microscopy.Com} ; Wed, 21 Jun 2006 22:34:37 -0500 21, 32 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; 21, 32 -- s=dk20050327; d=sprynet.com; 21, 32 -- b=Xd/a6Vln7K7YcDyDRjxr7rahw0V65rGdYJkVgR8/2tw4AvfSEPsCF2sUI+xxbFg6; 21, 32 -- h=Received:Reply-To:From:To:Cc:Subject:Date:Organization:Message-ID:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Priority:X-MSMail-Priority:X-Mailer:X-MimeOLE:Importance:X-ELNK-Trace:X-Originating-IP; 21, 32 -- Received: from [4.224.249.140] (helo=yourw92p4bhlzg) 21, 32 -- by elasmtp-junco.atl.sa.earthlink.net with asmtp (TLSv1:RC4-MD5:128) 21, 32 -- (Exim 4.34) 21, 32 -- id 1FtFxi-0004ct-Pf; Wed, 21 Jun 2006 23:34:36 -0400 21, 32 -- Reply-To: {ph2-at-sprynet.com} 21, 32 -- From: "Tony Havics" {ph2-at-sprynet.com} 21, 32 -- To: "Microscopy Listserve" {Microscopy-at-Microscopy.Com} 21, 32 -- Cc: {cloverbags-at-yahoo.com} 21, 32 -- Subject: pH of Oils 21, 32 -- Date: Wed, 21 Jun 2006 23:39:37 -0500 21, 32 -- Organization: pH2 21, 32 -- Message-ID: {002901c695b5$e0e0dcd0$8cf9e004-at-QEPI.local} 21, 32 -- MIME-Version: 1.0 21, 32 -- Content-Type: text/plain; 21, 32 -- charset="UTF-8" 21, 32 -- X-Priority: 3 (Normal) 21, 32 -- X-MSMail-Priority: Normal 21, 32 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 21, 32 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 21, 32 -- Importance: Normal 21, 32 -- X-ELNK-Trace: 6888e50b2be9b4fee5331016acda17f9bccc22a0f0e71d8e2dcc2d4da497cf5a350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c 21, 32 -- X-Originating-IP: 4.224.249.140 21, 32 -- Content-Transfer-Encoding: 8bit 21, 32 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5M3Yano024776 ==============================End of - Headers==============================
Greetings, It is likely that the pH of any oil is 7, almost by definition. I am sure a real chemist will correct me but pH expresses the relative abundance of protons over hydroxyls. As oils are non polar, they presumably repel a positive and negative charge equally and hence the pH is 'neutral', or a value of 7. Furthermore, the whole pH system is developed for aqueous solutions and there is presumably very little water indeed in the cedar oil, so it may not even be sensible to speak of pH for oils. Presumably that is why you can't find values in MSDSs, ehgo. Perhaps there is some solid-state notion of 'proton activity' that applies? Again maybe a chemist or physicist will have a more rigorous answer. Why do you care about the pH of these oils?
Hope this helps, Tobias } } Regarding: } } mail: cloverbags-at-yahoo.com } Name: raffaela } } Organization: cebu doctors' university } } Education: Undergraduate College } } Location: Cebu City, Philippines,Asia } } Question: what is the pH of a cedarwood } oilJuniperus virginiana) for oil immersion } objective? what is the pH of lemongrass } oil(Cymbopogon citratus)? } } 1. There are three types of cedar oil: } } Virginia cedarwood oil (Juniperus virginiana), } Texas cedarwood oil (Juniperus ashei or } mexicana), and Western red cedar (Thuja plicata) } } I have cedar oil (Texas) from Polysciences. } Their MSDS does not list a pH. Nor does Merck } list one. The Sigma Aldrich MSDS does not list } one, nor does the Libety Nautral Products } (Virginia), nor does JT Baker, nor does Well, } Naturally Products (Virginia & Texas & Western). } } I will try to get a pH on my Texas cedar oil } next week and report this at least. } } The constituents are mainly cedrene (a terpene) and cedral (cedar camphor). } } 2. regarding lemongrass oil: } } I have found no pH. Well, Naturally Products does not have it in their MSDS. } } Constituents (as listed by Merck): 75-85% } citral, methylheptenone, citronellel, geraniol, } limonene } } 3. I have a client that extracts, sells, } distributes these oils and will check for their } data as well next week. } } Tony } } .......................................................................... } Andrew Anthony "Tony" Havics, CHMM, CIH, PE } pH2, LLC } PO Box 34140 } Indianapolis, IN 46234 } (317) 752-6386 } (317) 409-3238 cell } } 90% of Risk Management is knowing where to place } the decimal point...any consultant can give you } the other 10%’ÑÝ } } This message is from pH2. This message and any } attachments may contain legally privileged or } confidential information, and are intended only } for the individual or entity identified above as } the addressee. If you are not the addressee, or } if this message has been addressed to you in } error, you are not authorized to read, copy, or } distribute this message and any attachments, and } we ask that you please delete this message and } attachments (including all copies) and notify } the sender by return e-mail or by phone at } 317-752-6386. Delivery of this message and any } attachments to any person other than the } intended recipient(s) is not intended in any way } to waive confidentiality or a privilege. All } personal messages express views only of the } sender, which are not to be attributed to pH2 } and may not be copied or distributed without } this statement. } } } } ==============================Original Headers============================== } 21, 32 -- From ph2-at-sprynet.com Wed Jun 21 22:34:37 2006 } 21, 32 -- Received: from } elasmtp-junco.atl.sa.earthlink.net } (elasmtp-junco.atl.sa.earthlink.net } [209.86.89.63]) } 21, 32 -- by ns.microscopy.com } (8.12.11.20060308/8.12.8) with ESMTP id } k5M3Yano024776 } 21, 32 -- for {Microscopy-at-Microscopy.Com} ; } Wed, 21 Jun 2006 22:34:37 -0500 } 21, 32 -- DomainKey-Signature: a=rsa-sha1; q=dns; c=nofws; } 21, 32 -- s=dk20050327; d=sprynet.com; } 21, 32 -- } b=Xd/a6Vln7K7YcDyDRjxr7rahw0V65rGdYJkVgR8/2tw4AvfSEPsCF2sUI+xxbFg6; } 21, 32 -- } h=Received:Reply-To:From:To:Cc:Subject:Date:Organization:Message-ID:MIME-Version:Content-Type:Content-Transfer-Encoding:X-Priority:X-MSMail-Priority:X-Mailer:X-MimeOLE:Importance:X-ELNK-Trace:X-Originating-IP; } 21, 32 -- Received: from [4.224.249.140] (helo=yourw92p4bhlzg) } 21, 32 -- by } elasmtp-junco.atl.sa.earthlink.net with asmtp } (TLSv1:RC4-MD5:128) } 21, 32 -- (Exim 4.34) } 21, 32 -- id 1FtFxi-0004ct-Pf; Wed, 21 Jun 2006 23:34:36 -0400 } 21, 32 -- Reply-To: {ph2-at-sprynet.com} } 21, 32 -- From: "Tony Havics" {ph2-at-sprynet.com} } 21, 32 -- To: "Microscopy Listserve" {Microscopy-at-Microscopy.Com} } 21, 32 -- Cc: {cloverbags-at-yahoo.com} } 21, 32 -- Subject: pH of Oils } 21, 32 -- Date: Wed, 21 Jun 2006 23:39:37 -0500 } 21, 32 -- Organization: pH2 } 21, 32 -- Message-ID: {002901c695b5$e0e0dcd0$8cf9e004-at-QEPI.local} } 21, 32 -- MIME-Version: 1.0 } 21, 32 -- Content-Type: text/plain; } 21, 32 -- charset="UTF-8" } 21, 32 -- X-Priority: 3 (Normal) } 21, 32 -- X-MSMail-Priority: Normal } 21, 32 -- X-Mailer: Microsoft Outlook, Build 10.0.2627 } 21, 32 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 } 21, 32 -- Importance: Normal } 21, 32 -- X-ELNK-Trace: } 6888e50b2be9b4fee5331016acda17f9bccc22a0f0e71d8e2dcc2d4da497cf5a350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c350badd9bab72f9c } 21, 32 -- X-Originating-IP: 4.224.249.140 } 21, 32 -- Content-Transfer-Encoding: 8bit } 21, 32 -- X-MIME-Autoconverted: from } quoted-printable to 8bit by ns.microscopy.com id } k5M3Yano024776 } ==============================End of - Headers==============================
This Question/Comment was submitted to the Microscopy Listserver using the WWW based Form at http://www.microscopy.org/MicroscopyListserver/MLFormMail.html --------------------------------------------------------------------------- Remember this posting is most likely not from a Subscriber, so when replying please copy both rpowell-at-nanoprobes.com as well as the MIcroscopy Listserver ---------------------------------------------------------------------------
Email: rpowell-at-nanoprobes.com Name: Rick Powell
Not sure how flexible you are in terms of your staining and preparation procedure... a PubMed search with the terms "post-embedding AND gold AND peroxidase" gave me 35 results, citing a variety of combinations of methods.
We have encountered several papers where silver-enhanced Nanogold (disclaimer - Nanogold is our product!) was used with DAB for double, or in the most recent case triple immunolabeling. We have summarized a couple of these in our newsletter - here are the references and links:
(1) Triple labeling, pre-embedding procedure:
Feng, Y.-P.; Li, Y.-Q.; Wang, W.; Wu, S.-X.; Chen, T.; Shigemoto, R., and Mizuno, N.: Morphological evidence for GABA/glycine-cocontaining terminals in synaptic contact with neurokinin-1 receptor-expressing neurons in the sacral dorsal commissural nucleus of the rat. Neurosci. Lett., 18, 144-148.
Reference describing methods in more detail:
Li, J. L.; Wang, D.; Kaneko, T.; Shigemoto, R.; Nomura, S., and Mizuno, N.: Relationship between neurokinin-1 receptor and substance P in the striatum: light and electron microscopic immunohistochemical study in the rat. J. Comp. Neurol., 418, 156ñ163 (2000).
Our article:
http://www.nanoprobes.com/Vol6_Iss11.html#3
(2) Pre-embedding localization of m2R and either ChAT or VAChT in the nucleus basalis magnocellularis (NBM) in rat brain sections where m2R was detected using pre-embedding ultrasmall (0.8 nm) gold enhanced with HQ Silver, and ChAT or VAChT by pre-embedding immunoperoxidase using the peroxidase anti-peroxidase (PAP) technique:
Decossas, M.; Doudnikoff, E.; Bloch, B., and Bernard, V.: Aging and subcellular localization of m2 muscarinic autoreceptor in basalocortical neurons in vivo. Neurobiol. Aging, 26, 1061-1072 (2005).
Our article:
http://www.nanoprobes.com/Vol6_Iss3.html#5
(3) Similar (pre-embedding labeling of vesicular glutamate transporter 1 (VGLUT1) and choline acetyltransferase (ChAT) immuno-reactivity respectively in the rat lumbar spinal cord):
Wu, S.-X.; Koshimizu, Y.; Feng, Y. P.; Okamoto, K.; Fujiyama, F.; Hioki, H.; Li, Y. Q.; Kaneko, T., and Mizuno, N.: Vesicular glutamate transporter immunoreactivity in the central and peripheral endings of muscle-spindle afferents. Brain Res.,} 1011, 247-251 (2004).
Other pre-embedding method:
Gutekunst, C. A.; Torre, E. R.; Sheng, Z.; Yi, H.; Coleman, S. H.; Riedel, I. B., and Bujo, H.: Stigmoid Bodies Contain Type I Receptor Proteins SorLA/LR11 and Sortilin. New perspectives on their function. J. Histochem. Cytochem., 51, 841-852 (2003).
Our article:
http://www.nanoprobes.com/Vol5_Iss7.html#4
Hope some of these are helpful,
Rick Powell Nanoprobes, Incorporated www.nanoprobes.com
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Hi All, I have been trying unsuccessfully to immuno-gold label HUVEC cells that have been embedded in LR White resin. The cells were plated and grown to supposed confluence, incubated with primary antibodies to PECAM and another adherence molecule, then fixed in 2% pfa, 0.5% glut., dehydrated, & embedded. The resin was cured at 50 deg. C overnight. En face sections were hydrated & blocked with buffer, then incubated with gold-tagged secondary Ab (anti-mouse or anti-rabbit). The sections were contrasted with aqueous Ur. Ac. then Pb citrate. I"ve seen nothing. For secondaries, I tried both F(ab)2s and whole IgG. I have been very successful with the same F(ab)2 secondaries on other samples processed the same way, but on which both primary and secondary Ab incubations were done on grid. In the past, we have successfully localized each of these primary Abs with peroxidase-DAB labelling prior to stringent fixation and embedding in Spurr's resin.
One of the problems is that HUVECs (human umbilical cord endothelial cells) are so darned flat...(less than 0.3 micrometers except for the nucleus) that its easy to cut right through them just trying to get a smooth section with no holes off of the raw block face. Another problem is that the proteins of interest are located at the cell junctions, in little vesicles and its not always easy to find cell-cell contacts.
My question (finally) is this: Has anyone tried to do an immuno-gold approach after immuno-peroxidase? We thought we'd like to try repeating the successful labelling of PECAM with DAB, pre-embedding, then try for the other Ab post-embedding.
Any ideas? (I'm sure there are....) thanks in advance, Lee
The histotechnologist in our group is having difficulty keeping formalin-fixed, paraffin-embedded sections of pig hooves on the slide for staining. Does anyone have any suggestions for her? She uses superfrost slides which works for all other tissues. If you have any hints for changes in fixation, etc. please pass those along.
Thanks! Peggy Peggy Sherwood Lab Associate, Photopathology Wellman Center for Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
==============================Original Headers============================== 6, 23 -- From MSHERWOOD-at-PARTNERS.ORG Thu Jun 22 09:31:02 2006 6, 23 -- Received: from PHSXCON5.partners.org (phsxcon5.mgh.harvard.edu [132.183.130.38]) 6, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MEV2ls001555 6, 23 -- for {Microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 09:31:02 -0500 6, 23 -- Received: from PHSXMB1.partners.org ([132.183.118.117]) by PHSXCON5.partners.org with Microsoft SMTPSVC(6.0.3790.211); 6, 23 -- Thu, 22 Jun 2006 10:31:00 -0400 6, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5.7226.0 6, 23 -- Content-class: urn:content-classes:message 6, 23 -- MIME-Version: 1.0 6, 23 -- Content-Type: text/plain; 6, 23 -- charset="iso-8859-1" 6, 23 -- Subject: FW: [LM Microsocpy] Re: staining of animal hooves and/or nails 6, 23 -- Date: Thu, 22 Jun 2006 10:31:00 -0400 6, 23 -- Message-ID: {1AF23D0AD12E7444A5DB083CA978B73407A309DD-at-PHSXMB1.partners.org} 6, 23 -- X-MS-Has-Attach: 6, 23 -- X-MS-TNEF-Correlator: 6, 23 -- Thread-Topic: [LM Microsocpy] Re: staining of animal hooves and/or nails 6, 23 -- Thread-Index: AcaWCBNPN2zboBv3S4eAdaMG2lZvqwAAFWkQ 6, 23 -- From: "Sherwood, Margaret " {MSHERWOOD-at-PARTNERS.ORG} 6, 23 -- To: {Microscopy-at-microscopy.com} 6, 23 -- X-OriginalArrivalTime: 22 Jun 2006 14:31:00.0271 (UTC) FILETIME=[7B82C3F0:01C69608] 6, 23 -- Content-Transfer-Encoding: 8bit 6, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MEV2ls001555 ==============================End of - Headers==============================
I was under the impression that, although oils can contain acidic contaminates, these are often removed during the refining process (not being good for engines etc.). Acidity (the acid-type constituents) "is usually defined in terms of total acid number (TAN). These constituents vary in nature and may or may not markedly influence the behaviour of the lubricant". Low acidity oils can oxidise during use and become more acidic over time (years) - e.g. the acid number goes to over 0.4 - while the newly refined 'non-corrosive' oil would be expected to be nearer to 0.05. The acids can be weak organic and strong inorganic. Acidity in oil can be useful for things like improved water resistance. Oils don't have an aqueous pH as such though, just the TAN number (total acidic number).
Olive oil for example is also graded for consumption on it's oleic acid acidity - around 3% in cheaper olive oil, and down to below 1% in 'extra virgin'. Oils are generally insoluble in water, which rather helps reduce it's corrosive properties during use.
Cargille use the Neutralization Number for their immersion oils. This is a measure of the acidity or alkalinity of the oil and is the mass in milligrams of the amount of acid (HCl) or base (KOH) required to neutralize one gram of the oil. For their standard immersion oils and type FF the value is very low [acidity] at 0.01 [KOH] (whereas the type DF is a rather high at 0.15 and the bottle states that it may damage objectives in constant use - it has lower auto-fluorescence than FF though). Cargille states that Cedarwood oil has a KOH neutralisation number greater than these values (ie. it is more acidic).
When used for objective immersion, the main things of interest in an oil would be things like : refractive index, colour, fluorescent properties, likely damage to the lens cement, crystallisation when cold (reversible), and whether the oil evaporates leaving a hard residue. The metal outside parts of the objective generally have an anti-corrosion finish, although here with our inverted microscopes spillage of corrosive culture media has sometimes caused some damage at the base of the objective, in particular corrosion of the area around the screw thread. The immersion oils generally dissolve in things like ethyl alcohol. Rather like engine oils for your car one tends to stick with the manufacturers [typically expensive] recommended one and not look too deeply into it's chemistry (microscope manufacturer's will soak test their objectives immersed for 6 months or more in an immersion oil before recommending it).
Oil acids can attack the lens mounting cement [see Cargille's comments below]. Plus the organic solvents used to remove lens immersion oil (like Xylene and ethanol) have been implicated in lens cement damage - I use ether generally for cleaning (and only clean rarely, when the lens is too dirty to be cleaned by just a lens tissue). Modern [low acid content] immersion oils supplied by the microscope manufacturer seem to be safe for longer term contact with the objective and don't evaporate to a hard residue (unlike say corn oil).
Cargille quote immersion oil shelf lives of 10 years (FF) and 5 years (DF) - and half that when opened.
On Cargille's site [http://www.cargille.com/immersionoilmicroscope.shtml] John Cargille states:
"To the knowledge of the author, no immersion oil matches the optics of the microscope perfectly. One of the closest matches is thickened cedarwood oil which, for many years, was the most widely used, if not the only immersion oil available. The disadvantageous properties of cedarwood oil are: high absorption of blue and UV light, yellowing with age, a tendency to harden on lenses due to uneven volatility, acidity, and changing viscosity (diluting with solvent changes the index and dispersion). The synthetic immersion oils eliminate many of the disadvantages cited above.
The acid value of immersion oil should be very low. The synthetics usually have acid values lower than cedarwood oil.
High acidity can, in time, affect the condition of the metal parts of the objective, and possibly more important, can cause deterioration of lens cements. The lens cement is also a seal that prevents oil from penetrating to the back of the lens. A crack or perforation in the cement draws oil by capillary attraction and a thin film of oil slowly creeps over the back of the objective lens; a poor image may develop without being immediately noticed. When the microscopist realizes the image has deteriorated, he may not realize the cause unless he has faced this problem before."
See Cargille's excellent site for more immersion oil info.
Regards
Keith
---------------------------------------------------------- Dr Keith J Morris Imaging Facilities Manager Cell Biology Division Institute of Ophthalmology University College London 11-43 Bath Street London EC1V 9EL
-----Original Message----- X-from: ph2-at-sprynet.com [mailto:ph2-at-sprynet.com] Sent: 22 June 2006 04:40 To: keith.morris-at-ucl.ac.uk
Regarding:
mail: cloverbags-at-yahoo.com Name: raffaela
Organization: cebu doctors' university
Education: Undergraduate College
Location: Cebu City, Philippines,Asia
Question: what is the pH of a cedarwood oilJuniperus virginiana) for oil immersion objective? what is the pH of lemongrass oil(Cymbopogon citratus)?
1. There are three types of cedar oil:
Virginia cedarwood oil (Juniperus virginiana), Texas cedarwood oil (Juniperus ashei or mexicana), and Western red cedar (Thuja plicata)
I have cedar oil (Texas) from Polysciences. Their MSDS does not list a pH. Nor does Merck list one. The Sigma Aldrich MSDS does not list one, nor does the Libety Nautral Products (Virginia), nor does JT Baker, nor does Well, Naturally Products (Virginia & Texas & Western).
I will try to get a pH on my Texas cedar oil next week and report this at least.
The constituents are mainly cedrene (a terpene) and cedral (cedar camphor).
2. regarding lemongrass oil:
I have found no pH. Well, Naturally Products does not have it in their MSDS.
3. I have a client that extracts, sells, distributes these oils and will check for their data as well next week.
Tony
.......................................................................... Andrew Anthony "Tony" Havics, CHMM, CIH, PE pH2, LLC PO Box 34140 Indianapolis, IN 46234 (317) 752-6386 (317) 409-3238 cell
90% of Risk Management is knowing where to place the decimal point...any consultant can give you the other 10%â„
==============================Original Headers============================== 43, 27 -- From keith.morris-at-ucl.ac.uk Thu Jun 22 09:52:39 2006 43, 27 -- Received: from vscani-d.ucl.ac.uk (vscani-d.ucl.ac.uk [144.82.108.132]) 43, 27 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MEqdvX011845 43, 27 -- for {Microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 09:52:39 -0500 43, 27 -- Received: from cell23.ioo.ucl.ac.uk ([144.82.134.123] helo=keithhigrade) 43, 27 -- by vscani-d.ucl.ac.uk with esmtp (Exim 4.60) 43, 27 -- (envelope-from {keith.morris-at-ucl.ac.uk} ) 43, 27 -- id 1FtQXs-0002j4-SV; Thu, 22 Jun 2006 15:52:37 +0100 43, 27 -- From: "Keith J Morris" {keith.morris-at-ucl.ac.uk} 43, 27 -- To: {Microscopy-at-microscopy.com} 43, 27 -- Cc: {cloverbags-at-yahoo.com} 43, 27 -- Subject: RE: [Microscopy] pH of Oils 43, 27 -- Date: Thu, 22 Jun 2006 15:52:33 +0100 43, 27 -- Message-ID: {000001c6960b$7ea22500$7b865290-at-keithhigrade} 43, 27 -- MIME-Version: 1.0 43, 27 -- Content-Type: text/plain; 43, 27 -- charset="iso-8859-1" 43, 27 -- X-Mailer: Microsoft Office Outlook 11 43, 27 -- Thread-Index: AcaVrZneujlpeoxMTIOEKedyM/LyNwATipAg 43, 27 -- In-Reply-To: {200606220340.k5M3eN8d031970-at-ns.microscopy.com} 43, 27 -- X-MimeOLE: Produced By Microsoft MimeOLE V6.00.2900.2869 43, 27 -- X-UCL-MailScanner-Information: Please contact the UCL Helpdesk, helpdesk-at-ucl.ac.uk for more information 43, 27 -- X-UCL-MailScanner: Found to be clean 43, 27 -- X-UCL-MailScanner-From: keith.morris-at-ucl.ac.uk 43, 27 -- X-Spam-Status: No 43, 27 -- Content-Transfer-Encoding: 8bit 43, 27 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MEqdvX011845 ==============================End of - Headers==============================
We have always used ethanol as the dehydration solvent with our Balzers CPD 020. Does anyone know whether the seals and other components of this instrument are compatible with methanol? It is used as a primary fixative for plant tissue in a publication, but it is unclear from the methods section whether the tissue is changed to ethanol before critical point drying. We have sent e-mails to the author and US supplier of Balzers equipment, but not yet received any replies. Also, does anyone know whether absolute ethanol can be directly changed for anhydrous methanol without damaging plant tissue?
Heather Owen
Dr. Heather A. Owen, Director Electron Microscope Laboratory Department of Biological Sciences University of Wisconsin - Milwaukee Lapham Hall 3209 N. Maryland Ave. Milwaukee, WI 53211 USA
Phone: (414)229-6816
==============================Original Headers============================== 7, 19 -- From owenha-at-csd.uwm.edu Thu Jun 22 11:33:17 2006 7, 19 -- Received: from batch3.csd.uwm.edu (batch3.csd.uwm.edu [129.89.169.226]) 7, 19 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MGXGdi023787 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 11:33:17 -0500 7, 19 -- Received: from alpha2.csd.uwm.edu (alpha2.csd.uwm.edu [129.89.7.202] (may be forged)) 7, 19 -- by batch3.csd.uwm.edu (8.13.6/8.12.6) with ESMTP id k5MGXEav030866 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 11:33:14 -0500 (CDT) 7, 19 -- Received: from localhost (owenha-at-localhost) 7, 19 -- by alpha2.csd.uwm.edu (8.13.6/8.12.6) with SMTP id k5MGXEOd004137 7, 19 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 11:33:14 -0500 (CDT) 7, 19 -- X-Authentication-Warning: alpha2.csd.uwm.edu: owenha owned process doing -bs 7, 19 -- Date: Thu, 22 Jun 2006 11:33:14 -0500 (CDT) 7, 19 -- From: Heather A Owen {owenha-at-csd.uwm.edu} 7, 19 -- Reply-To: Heather A Owen {owenha-at-csd.uwm.edu} 7, 19 -- To: microscopy-at-microscopy.com 7, 19 -- Subject: Balzers Critical Point Drier solvents 7, 19 -- Message-ID: {Pine.OSF.3.96.1060622111531.10763A-100000-at-alpha2.csd.uwm.edu} 7, 19 -- MIME-Version: 1.0 7, 19 -- Content-Type: TEXT/PLAIN; charset=US-ASCII ==============================End of - Headers==============================
We have been polymerizing our excess resins from infiltrations, etc., in the same oven we use for polymerizing our actual samples in resin. My question is whether this could have an effect on the quality of the resin blocks, maybe due to the solvents in the leftovers?
We had a couple batches of semi-gummy bears recently and this question has been resinating around the lab ever since. No samples lost, but a definite annoyance.
Thanks for any thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Thu Jun 22 12:52:34 2006 10, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MHqYXe002752 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 12:52:34 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: TEM: polymerization question 10, 23 -- Date: Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- Message-ID: {91108EF9255B394CBF8B7E3789814A411E332E-at-UM-XMAIL08.um.umsystem.edu} 10, 23 -- X-MS-Has-Attach: 10, 23 -- X-MS-TNEF-Correlator: 10, 23 -- Thread-Topic: TEM: polymerization question 10, 23 -- thread-index: AcaWJKO/b+LSDKGdS4WFWVdZe1OTcA== 10, 23 -- From: "Tindall, Randy D." {TindallR-at-missouri.edu} 10, 23 -- To: {microscopy-at-microscopy.com} 10, 23 -- X-OriginalArrivalTime: 22 Jun 2006 17:52:34.0055 (UTC) FILETIME=[A3F7C170:01C69624] 10, 23 -- Content-Transfer-Encoding: 8bit 10, 23 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id k5MHqYXe002752 ==============================End of - Headers==============================
First a question: Is the oven in a hood? And is the oven ventilated on top (most of them I've run across either have pluggable holes or some sort of vane type shutter on top.
I haven't noticed any issue polymerizing with alcohol contaminated resin in the oven at the same time as the blocks. Although more often than not I will let the waste sit out in the hood until the blocks are cured and then stick in the waste to cure until next time I'm ready to put a batch in.
HTH
Geoff Williams Leduc Bioimaging Facility Manager Brown University
-----Original Message----- X-from: TindallR-at-missouri.edu [mailto:TindallR-at-missouri.edu] Sent: Thursday, June 22, 2006 1:57 PM To: Williams, Geoffrey
Hi all,
We have been polymerizing our excess resins from infiltrations, etc., in the same oven we use for polymerizing our actual samples in resin. My question is whether this could have an effect on the quality of the resin blocks, maybe due to the solvents in the leftovers?
We had a couple batches of semi-gummy bears recently and this question has been resinating around the lab ever since. No samples lost, but a definite annoyance.
Thanks for any thoughts.
Randy
Randy Tindall EM Specialist Electron Microscopy Core Facility---We Do Small Well! W125 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-2227 Email: tindallr-at-missouri.edu Web: http://www.emc.missouri.edu
==============================Original Headers============================== 10, 23 -- From TindallR-at-missouri.edu Thu Jun 22 12:52:34 2006 10, 23 -- Received: from um-tsmtpout1.um.umsystem.edu (um-tsmtpout1.um.umsystem.edu [209.106.228.23]) 10, 23 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id k5MHqYXe002752 10, 23 -- for {microscopy-at-microscopy.com} ; Thu, 22 Jun 2006 12:52:34 -0500 10, 23 -- Received: from UM-XMAIL08.um.umsystem.edu ([209.106.228.34]) by um-tsmtpout1.um.umsystem.edu with Microsoft SMTPSVC(6.0.3790.1830); 10, 23 -- Thu, 22 Jun 2006 12:52:33 -0500 10, 23 -- X-MimeOLE: Produced By Microsoft Exchange V6.5 10, 23 -- Content-class: urn:content-classes:message 10, 23 -- MIME-Version: 1.0 10, 23 -- Content-Type: text/plain; 10, 23 -- charset="us-ascii" 10, 23 -- Subject: TEM: polymerization question 10, 23 -- Date: T