Precise language with universally accepted definitions form the basis of rational scientific discussion. If this is pedantry, it seems to be warranted here.
Betty Abajian-Seaman Wyeth Pearl River, NY
} } } Malcolm Haswell {malcolm.haswell-at-sunderland.ac.uk} 4/30/2003 4:32:28 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
There have been several listers who seem to be calling graticules and stage micrometers the same thing. In my experience in UK light microscopy a stage micrometer is a specific accurate 'ruler' which may be mounted on a light microscope slide. It is an absolute size such as 1 or 10mm and is subdivided for accurate absolute measurements. Most graticules are eyepiece devices which may have up to 100 linear divisions or be divided up in some other way such as by angles. You can calibrate an eyepiece graticule using a stage micrometer for a particular objective lens configuration but the graticule is still an arbitrary measure.
I suppose you could argue that a stage micrometer was a specific absolute graticule but that does not make the terms interchangeable. If I am being pedantic, I apologise.
Thanks for your attention
Malcolm Haswell e.m. unit University of Sunderland UK
Hi Karen, We have been using Xylene/Oso4 for many years.It works quickly and with good results if original fixation was buffered formalin. The reference is as follows: Kai Chien,R.L.Van deVeldeand R.C.Hauser, Fortieth Annual EMSA Meeting,1982,pp356-357 I don't know if it was ever published any where else. We use .01grm of OsO4 rather than using a whole grm,since we only do it a few times a year. Our procedure is as follows: two 20 minute changes in xylene one 20 minute change in xylene/OsO4 two 10 minute changes in Propylene oxide 15 minutes in 3:1 P.O./resin mixture 30 minutes 1:1 " " " 30 minutes 1:3 " " " 20 minutes in fresh resin-- embed
Almost easier than starting with fresh tissue! Cheers, Connie
-----Original Message----- } From: "kbovard-at-creighton.edu"-at-sparc5.microscopy.com [mailto:"kbovard-at-creighton.edu"-at-sparc5.microscopy.com] Sent: Wednesday, April 30, 2003 12:23 PM To: Microscopy-at-sparc5.microscopy.com
Does anyone have any information about whether osmium tetroxide can be used in xylene?
In our clinical pathology EM lab I am frequently called upon to *rescue* a piece of tissue from a paraffin block and work it up for EM. The standard protocol is to deparaffinate the tissue in xylene, then hydrate the tissue to osmicate it then dehydrate it again to go on to infiltrating with resin and finally embedding/polymerizing.
I have a faint recollection of a discussion years ago that osmium could be suspended in xylene saving the whole hydration/osmication/dehydration step. Thus one would deparaffinate the tissue in xylene then osmicate in xylene and go on to infiltration from there.
Any comments?
Karen Bovard Creighton University Medical Center Department of Pathology Electron Microscopy Laboratory Omaha, Nebraska
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (WOwsiany-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, April 30, 2003 at 11:59:57 ---------------------------------------------------------------------------
Email: WOwsiany-at-aol.com Name: Walter J. Owsiany
Organization: Vineyards Country Club
Education: Undergraduate College
Location: Naples, Florida 34119
Question: I am a golf course superintendent, what type of scope would I need to look at turfgrasses diseases in my office. Spending no more than $8oo to $1000.
Amanda, We convinced our IT dept. to install SNAP servers to accommodate the images generated by our Optical Microscopy Core Facility, and the rest of the department. It has worked out quite well. Lee
} -----------------------------------------------------------------------. } } } Hello all, } I am trying to gather information about saving digital images. } Currently I import all images to Quartz PCI and save them as TIFF files on } our network. I would like to continue to do this however, the IT department } is threatening to cut me off if I continue to use so much space on the } server. We do occasional image analysis, and since JPEG compression is lossy } and introduces artifacts, I am leary of heading in that direction. } I thought that if I could convince them that this is the best practice for } saving images we could come to some sort of agreement. Any information of } how you handle saving SEM images would be helpful in my quest for peace. } Thank you } } ~Amanda Marchese } } } PQ Corporation } Research & Development Center } 280 Cedar Grove Road } Conshohocken PA 19428 } (610) 651-4668
-- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
I concur. We are a pathology lab for a research group, and that is standard procedure: fixing a piece of each for LM and EM. It's a lot of work to deparaffinize and re-fix a piece of tissue for such poor results! However, sometimes it's unavoidable, if the tissue is archival and now they need to look at ultrastructure.
I do so few retrieval samples, that I was interested to learn about mixing osmium in xylene. Thanks for the information!
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of 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 Message----- } From: Garrison, Becky [SMTP:becky.garrison-at-jax.ufl.edu] } Sent: Wednesday, April 30, 2003 4:18 PM } To: '"kbovard-at-creighton.edu"-at-sparc5.microscopy.com'; } Microscopy-at-sparc5.microscopy.com } Subject: RE: Osmium in xylene } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Yes. Osmium dissloves in xylene. I have used a similar procedure. Biggest } problem is measuring a tiny amount of osmium, since such a small amount is } needed. Our Mettler is not in a hood, only an old swinging arm balance. I } keep one vial of OsO4 crystals for such use. } On the whole, over the years, such "rescued" tissue does not give good } results and in most cases the results are uninterpretable and } non-diagnostic. } It is much more effective to put a snip of tisue for any case which might } possibly need EM in EM fixative for potential use. The cost of processing } one case for EM with poor results far outweighs the cost of a batch (or 2) } of EM fixative and vials. } } Becky Garrison } Pathology Supervisor } } } -----Original Message----- } } From: "kbovard-at-creighton.edu"-at-sparc5.microscopy.com } [mailto:"kbovard-at-creighton.edu"-at-sparc5.microscopy.com] } Sent: Wednesday, April 30, 2003 12:23 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Osmium in xylene } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have any information about whether osmium tetroxide can be } used in xylene? } } In our clinical pathology EM lab I am frequently called } upon to *rescue* a piece of tissue from a paraffin block and work it up } for EM. The standard protocol is to deparaffinate the tissue in xylene, } then hydrate the tissue to osmicate it then dehydrate it again to go on to } infiltrating with resin and finally embedding/polymerizing. } } I have a faint recollection of a discussion years ago that osmium could be } suspended in xylene saving the whole hydration/osmication/dehydration } step. Thus one would deparaffinate the tissue in xylene then osmicate in } xylene and go on to infiltration from there. } } Any comments? } } } Karen Bovard } Creighton University Medical Center } Department of Pathology } Electron Microscopy Laboratory } Omaha, Nebraska
We also use PCI and, as you are aware, the big advantage of a database system is the availability of images. I certainly believe you are using a sound strategy and suggest you try to keep your images on the server. If you are not currently performing daily back-ups I would also suggest that you consider implementing a routine in order to protect against data loss.
Isn't it amazing? The IT department is charged with the difficult task of supporting an ever changing network environment. In this silicon world of flux they lose sight of their mission....to support the users of the network!!! Amanda, they work for you, or at least that's how it should work! My suggestion is to convince the IT department that it is in everyone's best interests if the IT "experts" assist with a network solution to the problem. They need to provide you with network storage space and a timely backup routine in order to protect your valuable research, which is much more costly that a few gigabits of hard drive space. Finally, I would avoid setting up a local server since it places much more responsibility on you now and, more importantly, in the future. Things change.....and look how difficult it is for the experts to keep things running! :)
Good luck, jr
John A. Robson Boehringer Ingelheim Pharmaceuticals, Inc. PO Box 368 900 Ridgebury Rd Ridgefield, CT 06877
-----Original Message----- } From: Amanda Marchese(RD) [mailto:Amanda.Marchese-at-PQCorp.com] Sent: Wednesday, April 30, 2003 1:16 PM To: 'Microscopy-at-MSA.Microscopy.Com'
Hello all, I am trying to gather information about saving digital images. Currently I import all images to Quartz PCI and save them as TIFF files on our network. I would like to continue to do this however, the IT department is threatening to cut me off if I continue to use so much space on the server. We do occasional image analysis, and since JPEG compression is lossy and introduces artifacts, I am leary of heading in that direction. I thought that if I could convince them that this is the best practice for saving images we could come to some sort of agreement. Any information of how you handle saving SEM images would be helpful in my quest for peace. Thank you
~Amanda Marchese
PQ Corporation Research & Development Center 280 Cedar Grove Road Conshohocken PA 19428 (610) 651-4668
On Wednesday, April 30, 2003, at 07:40 AM, Charles Murphy wrote:
} Hi: We are currently investigating various freeze sub and freeze } plunge systems and would appreciate any pros or cons people have } towards the different systems. Vendors feel free to email me with } your products and please include the prices. } Dear Charlie, If your budget will cover it, I strongly recommend getting a Vitrobot for plunge freezing; the cost is ~$70k. It provides a consistent, controlled environment and gives excellent results. I have no financial interest in FEI, which sells the Vitrobot; I'm just a satisfied customer. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
I'm a researcher, currently unaffiliated, and I hope to purchase a compound scope and a dissecting scope to use in my mycological work. I'll be studying mycorrhizae and endophytic fungi in grasses, and I need scopes that will allow me to examine fungal hyphae and spores, as well as grass inflorescences. (I have a master's in botany and will begin a Ph.D. in the fall of 2004, when I'll have access to high-quality equipment.)
I am looking for scopes that are durable (I'll be traveling with them to field stations), low maintenance, inexpensive (under $1000 each, and preferably less), and of course, adequate to the task of viewing fungi. Which brands and models would you recommend? Is it reasonable to buy used ones, say from ebay?
has anyone swapped out the 486DX2 that was common on many mid-late 90's 982s with a newer processor board. i have a PII 233 SBC that i'd like to install, but there seems to be a "timing" issue between it and the microscope hardware. any help would be appreciated...
Dennis Bellotto Research Scientist Pulmonary Research Mail code:9034 UT Southwestern Medical Center 5323 Harry Hines Blvd. Dallas, TX 75390-9034 ph:(214)648-3597/fax:(214)648-8027 pager: (214)822-0541 dennis.bellotto-at-utsouthwestern.edu Come to my aerobics classes: Dynastep Tue/Thur 5:20pm and Mon/Fri 12:15 at the Bryan Williams Center
Malcolm, that's always the way I learnt it, but then I also larned dis-section rather than di-section.
Being pedantic saddles one with the "p" word which is a lot better than being encumbered with either a "d" or "b" word.
Cheers,
Fred Monson
P.S. Speaking of reticles, reticules(#), graticles, graticules and mIcromEters, does anyone know where to find a 'reasonably' priced finder/locater (master) slide (square array of letters and numbers)? I purchased one from Scientific Products here in the USA some years back for around $50.00 and now the only ones I can find are closer to a very much bigger number. Any help would be appreciated. Apparently these things were all a lot less expensive to make when manufactured by hand by skilled technicians, but that was before automated lasers began turning them out. I guess now you don't have to send the locater slide with the specimen slide when you want to show your colleague where to look. Beer is more expensive and so are graticules, but now there is also a big market for brilliant yellow HummVeeez! Progress! Must be close to Friday.
-----Original Message----- } From: Malcolm Haswell [mailto:malcolm.haswell-at-sunderland.ac.uk] Sent: Wednesday, April 30, 2003 4:32 AM To: MSA microscopy listserver
There have been several listers who seem to be calling graticules and stage micrometers the same thing. In my experience in UK light microscopy a stage micrometer is a specific accurate 'ruler' which may be mounted on a light microscope slide. It is an absolute size such as 1 or 10mm and is subdivided for accurate absolute measurements. Most graticules are eyepiece devices which may have up to 100 linear divisions or be divided up in some other way such as by angles. You can calibrate an eyepiece graticule using a stage micrometer for a particular objective lens configuration but the graticule is still an arbitrary measure.
I suppose you could argue that a stage micrometer was a specific absolute graticule but that does not make the terms interchangeable. If I am being pedantic, I apologise.
Thanks for your attention
Malcolm Haswell e.m. unit University of Sunderland UK
We want to reduce/eliminate vibration associated with some water cooling lines on our TEM. One suggestion has been to mount the water coolers (Haskris) on a platform supported by truck tire inner tubes (to prevent ground transmission) and put some constrictions in the water line to reduce vibration transmission. The first one sounds good; I suspect that some sort of in-line filter would be better to remove vibrations in the water.
I would be interested in how others have approached this; I think it is a fairly general issue.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
I know it's not the same thing but we have a Hitachi S3000N scanning e.m. It is controlled by Windows NT 4.0 and we have been instructed that we shouldn't even install the latest service pack for the operating system because of potential conflicts especially with peripherals and the electron microscope controls.
What you are proposing sounds much more drastic and it might be worth getting advice from LEO or a service engineer. Have you got a particular reason for the upgrade because any possible gains might be far outweighed by the problems you may get? I suspect you would definitely be in the area of "invalidating the manufacturer's warranty".
I get the impression that many of the computerised facilities in modern instruments are tied down to particular hardware and/or software/firmware with little hope of upgrades to keep up with the pace of modern computing technologies. An obvious example might be if we need to upgrade to a new printer or removeable disc technology - my answer to date has been to network a modern computer to our system for archiving and driving newer technologies when we need them.
My concern is that it's not just the storage media (CD, DVD, magneto optical) on modern systems will be obsolete in a few years but the computing technologies (Operating systems, software and hardware) will be unsupported. We have used Win 95, 98 and NT 4.0 which I believe Microsoft is about to drop or already has dropped technical support for.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: brian mcintyre {mcintyre-at-optics.rochester.edu}
Dear Amanda,
One other note on image storage: the Quartz PCI database will keep track of the CDs or DVDs that you burn full of images, if you tell the database, and tell you to put in e.g. "CD #12345" when you want to access that image.
Regards, Mary Mager
Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Amanda Marchese(RD)" {Amanda.Marchese-at-PQCorp.com} To: "'Microscopy-at-MSA.Microscopy.Com'" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 30, 2003 10:16 AM
Dear Ritchie, I used to use the DOS code from John Donovan at Berkeley called CITZAF. I see there is an up-dated version of it on the MSA web site. Look for "Reference and Education Activities" on the bottom of the first page, then "Microscopy and Microanalysis Software Libraries". Under "03-MASlib/" you will find "CITZAF2" and "CITZAF3". They should help, although I haven't tried them myself. Good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca
----- Original Message ----- } From: "Ritchie Sims" {r.sims-at-auckland.ac.nz} To: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} ; "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 29, 2003 1:44 PM
I would like to thank those of you who have provided me with the information regarding add-on of the Z-motorized function on my Olympus BX51 microscope.
We are currently considering to go with Prior OptiScan ES101, which fits our purpose best.
Have a nice weekend.
Xinran Liu
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center at Dallas Phone: 214-648-1830 Fax: 214-648-1801 E-mail: xinran.liu-at-utsouthwestern.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (bop00rav-at-sheffield.ac.uk) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May 4, 2003 at 10:41:32 ---------------------------------------------------------------------------
Email: bop00rav-at-sheffield.ac.uk Name: robert vasey
Organization: university of sheffield
Education: Graduate College
Location: sheffield, uk
Question: I am fixing and cutting sections of plant roots for light microscopy. I am embedding in LR White resin. My question relates to fixation. Is it better to use sodium phosphate buffer or sodium cacodylate buffer? If one is better than the other could you tell me why.
We have someone who would like to examine osteoclast ruffled border formation on adult mouse vertebrae through TEM. We compared two protocols that were used on osteoclast study. From those protocols, we are planning to perfuse the mouse with 2% Formaldehyde (EM grade), 2.5% glutaraldehyde in 0.1M cacodylate buffer, followed by immersion fixation for 24 hours at room temp with the same fixative as the perfusate fixative but with 0.5% Calcium Chloride. Each vertebra is approximately 4mm x 2mm x 3mm in dimension. Although the recommended size of the sample for EM must be less than 1 mm in dimension, is the fixative concentration and length of fixation sufficient enough to fix the sample? It wasn't mentioned whether the samples were fixed for 24 hours either at room temp or in the refrigerator? Is fixation at room temp for 24 hours reasonable? Are there other means of cutting each vertebra into smaller dimensions for EM during the primary fixation? Following immersion fixation, the sample will be decalcified with 14% EDTA for 2 days. The samples will then be further cut into smaller dimensions, washed with cacodylate buffer, post-fixed with 2% osmium tetroxide for 1 hour, followed by gradual dehydration with ethanol, intermediate dehydration with propylene oxide, and gradual embedding with Glauert (Med) EMBED. We appreciate your suggestions. Thank you for your time.
Sincerely,
Claire Haueter
Claire Haueter EM Technician Integrated Microscopy Core Baylor College of Medicine 713-798-4952
I’m working with meosporous materials. I would like to get a computer simulation of the MCM41 structure with a multislice program. Can anyone give me some information about it?
Thanks
Emma.
Emma Rossinyol
Departament d'Electrònica Universitat de Barcelona Martí i Franqués 1 08028 Barcelona
Morning Walter, From what I can see, the great majority of turfgrass diseases can be observed by the naked eye or with a magnfiying glass, so for closer inspection, without any other need than identification or confirmation of identification, my suggestion would be for a dissecting microscope with either a ring light (fluorescent or fiber optic light) or directed light (separate or integrated) You must decide whether you need both incident and transmitted light, but it would appear that you should start with a scope such as the one on the web page below: http://www.wme-inc.com/WebPages/MetEqPgs/b&l4.htm others here: http://www.wme-inc.com/WebPages/MetEqPgs/Met01.htm#Microscopes
This used equipment is being sold by a company in Fort Walton Beach, but here is a company with reconditioned scopes and prices. http://www.midwestbioservicecompany.com/used.asp
B&L dissecting scopes are usually very good. I have had several for years.
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: WOwsiany-at-aol.com [mailto:WOwsiany-at-aol.com] Sent: Thursday, May 01, 2003 8:34 AM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (WOwsiany-at-aol.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, April 30, 2003 at 11:59:57 ---------------------------------------------------------------------------
Email: WOwsiany-at-aol.com Name: Walter J. Owsiany
Organization: Vineyards Country Club
Education: Undergraduate College
Location: Naples, Florida 34119
Question: I am a golf course superintendent, what type of scope would I need to look at turfgrasses diseases in my office. Spending no more than $8oo to $1000.
This is to thank everybody who replied with a wealth of ideas concerning Roots and SEM. If haven't had any feedback yet from our soil science department, so I can't as yet report any progress on that front.
However, in regard to moulds, I inadvertently (honest!) let some bread go mouldy recently. So I've looked through all the suggestions. For fixing, one can't slowly go up through the progressions of increasingly concentrated ethanol, since the bread will go intractably soggy (does anyone remember the song "Mouldy Old Dough"?) Neither do I want to hit the mould with neat alcohol. So what I'm doing, as a first stage at least, is to put thin mouldy crusts of the bread over methanol in a vapour tank, and letting it stand over the long weekend (we British are having our May Day Holiday on Monday 5th!). If anything useful results, I'll keep you posted.
Bye for now,
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +----------------------------------------+ Phone:+44 (0) 118 9318572 Fax: +44 (0) 118 9750203 University internal extension 7867 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +----------------------------------------+
Greetings, There is probably no absolute 'better'. The choice of reagent typically depends on what questions you want to ask, and hence what protocols you plan to follow.
It is worth knowing that cacodylate contains arsenic and so is quite toxic and must be handled and disposed with care. Generally, cacodylate buffers are used to avoid precipitation between phosphate and heavy metal, esp osmium. If you are doing light microscopy, your most likely won't be osmitcating, etc, and so the simple phosphate buffer should work fine. There may exist some specific protocols for LM that feature cacodylate. If you are going to do immuno localization, most of protocols for that call for fixation in an organic buffer, pipes, but I don't know that anyone has ever tested rigorously whether that is really better than phosphate. Good control of pH probably is needed.
Hope this helps, TObias
} } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (bop00rav-at-sheffield.ac.uk) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Sunday, May } 4, 2003 at 10:41:32 } --------------------------------------------------------------------------- } } Email: bop00rav-at-sheffield.ac.uk } Name: robert vasey } } Organization: university of sheffield } } Education: Graduate College } } Location: sheffield, uk } } Question: I am fixing and cutting sections of plant roots for light } microscopy. I am embedding in LR White resin. My question relates } to fixation. Is it better to use sodium phosphate buffer or sodium } cacodylate buffer? If one is better than the other could you tell } me why. } } Thanks. } } Bob. } } ---------------------------------------------------------------------------
Phosphate can lead to calcium precipitation which is noticeable in the TEM. Cacodylate doesn't have this problem but it is arsenic based and therefore toxic. As a hazardous waste, it needs proper disposal which can be expensive but more importantly, something that should be minimized unless absolutely necessary. I recommend HEPES or PIPES.
At 11:42 AM 5/4/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
The Materials Characterization Facility at the University of Pennsylvania is interested in selling a Phillips 420T TEM/STEM. The instrument has been maintained under service contract and is equipped with a Kevex 8204 Delta 1 EDS system and a Gatan serial electron energy loss spectrometer. The instrument comes with single-tilt and double-tilt, tilt-rotate, tensile, heating and cryogenic holders.
Additional information (specifications, picture, etc.) can be found at:
Please contact Doug Yates for further details regarding terms of sale at dmyates-at-lrsm.upenn.edu. The purchaser will be responsible for all relocation costs.
********************************************************* Douglas M. Yates, Ph.D. Technical Director Materials Characterization Facility 215-898-2013 dmyates-at-lrsm.upenn.edu Department of Materials Science & Engineering 3231 Walnut Street Philadelphia, PA 19104 *********************************************************
I used to do a lot of bones and teeth for EM, using a protocol similar to the one you describe but with some differences. I perfused rats with glutaraldehyde in phosphate buffer, then continued the fixation in Karnowski's fixative (paraformaldehyde and glutaraldehyde in cacodylate) for 24h at 4 degrees with continuous stirring. I was looking at pieces of rat skull much bigger than mouse vertebrae, and they seemed to be fixed properly. For the decalcification stage, you might want to check the end-point by x-ray, if possible, rather than giving all the samples a standard two days. Hope this helps.
Lesley Weston.
on 04/05/2003 8:10 PM, chaueter at chaueter-at-bcm.tmc.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } } We have someone who would like to examine osteoclast ruffled border } formation on adult mouse vertebrae through TEM. We compared two protocols } that were used on osteoclast study. From those protocols, we are planning to } perfuse the mouse with 2% Formaldehyde (EM grade), 2.5% glutaraldehyde in 0.1M } cacodylate buffer, followed by immersion fixation for 24 hours at room temp } with the same fixative as the perfusate fixative but with 0.5% Calcium } Chloride. Each vertebra is approximately 4mm x 2mm x 3mm in dimension. } Although the recommended size of the sample for EM must be less than 1 mm in } dimension, is the fixative concentration and length of fixation sufficient } enough to fix the sample? It wasn't mentioned whether the samples were fixed } for 24 hours either at room temp or in the refrigerator? Is fixation at room } temp for 24 hours reasonable? Are there other means of cutting each vertebra } into smaller dimensions for EM during the primary fixation? } Following immersion fixation, the sample will be decalcified with 14% EDTA } for 2 days. The samples will then be further cut into smaller dimensions, } washed with cacodylate buffer, post-fixed with 2% osmium tetroxide for 1 hour, } followed by gradual dehydration with ethanol, intermediate dehydration with } propylene oxide, and gradual embedding with Glauert (Med) EMBED. } We appreciate your suggestions. Thank you for your time. } } Sincerely, } } Claire Haueter } } Claire Haueter } EM Technician } Integrated Microscopy Core } Baylor College of Medicine } 713-798-4952 } }
There is a straight-foward and easy approach to eliminating vibrations that emanate from the chillers: Add an extra 10-20 feet of tubing between chiller and microscope, wind up the extra tubing and place on the floor (preferably somewhere out of the way). The coil of extra tubing absorbs the chiller vibration, thus solving the problem. This is an old solution that I have successfully used on several microscopes. Of course, it always helps to locate the chiller as far from the instrument as possible. Contact Haskris for this maximum distance.
I doubt that mounting the chiller on an anti-vibration platform will not work because you are only isolating the vibration from the floor, not from the tubing leading to and from the microscope.
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Cheers,
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"L. D. Marks" {ldm-at-risc4.numis.n To: Microscopy List wu.edu} {microscopy-at-sparc5.microscopy.com} cc: Subject: Antivibration measures in water lines (TEM) 05/02/03 06:24 AM
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We want to reduce/eliminate vibration associated with some water cooling lines on our TEM. One suggestion has been to mount the water coolers (Haskris) on a platform supported by truck tire inner tubes (to prevent ground transmission) and put some constrictions in the water line to reduce vibration transmission. The first one sounds good; I suspect that some sort of in-line filter would be better to remove vibrations in the water.
I would be interested in how others have approached this; I think it is a fairly general issue.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
A holding tank, and gravity feed of water to the microscopes is in use at the JEOL factory. Seems to work quite well. Inner tubes for floor vibration sounds like a great idea, but are truck tire inner tubes perhaps a bit big? I would have thought motorcycle inner tubes to be more like the right size, especially for stability of the platform..
John Mardinly Intel
-----Original Message----- } From: L. D. Marks [mailto:ldm-at-risc4.numis.nwu.edu] Sent: Friday, May 02, 2003 4:24 AM To: Microscopy List
We want to reduce/eliminate vibration associated with some water cooling lines on our TEM. One suggestion has been to mount the water coolers (Haskris) on a platform supported by truck tire inner tubes (to prevent ground transmission) and put some constrictions in the water line to reduce vibration transmission. The first one sounds good; I suspect that some sort of in-line filter would be better to remove vibrations in the water.
I would be interested in how others have approached this; I think it is a fairly general issue.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
We want to examine negative-stained liposomes that contain a protein inserted in the lipid membrane, using the TEM.
The technique we use for plant viruses is:
Place a drop of the suspension on parafilm Place a couple of formvar coated grids on the suspension for 5 minutes Blot Place grids on UA for 1-5 minutes Blot
Can we use the same technique for the liposomes? Are there tricks involved to maintain structural integrity and to avoid loss of specimens from the grid surface?
Thanks,
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
On Friday, May 2, 2003, at 04:24 AM, L. D. Marks wrote:
} We want to reduce/eliminate vibration associated with some } water cooling lines on our TEM. One suggestion has been to } mount the water coolers (Haskris) on a platform supported } by truck tire inner tubes (to prevent ground transmission) } and put some constrictions in the water line to reduce } vibration transmission. The first one sounds good; I suspect } that some sort of in-line filter would be better to remove } vibrations in the water. } } I would be interested in how others have approached this; } I think it is a fairly general issue. } Dear Laurence, In Albany, we added vibration dampers, which were cylinders with inflatable bladders inside. We maintained the bladders at about 2/3 the pressure of the water--which varied in the several locations and between the inflow and outflow lines. These dampers were recommended for a low-frequency range of vibrations, tens to a hundred or so Hz. They were mounted on T's off the water lines in locations near the Haskris, the lines to the lenses, the electronics racks and the camera DP. We also had in-line filters to keep the cooling water clean. Both types of installation were equipped with bypass valves, so we could measure the bladder pressure with and without water flow to the damper or the flow rate with and without the filter. This way we could maintain the system, which we did at regular intervals. I hope there is still someone at Albany who could give you more detailed info, such as the manufacturers, costs, etc. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
A vacancy for a technical assistant in the Structural Virology Group of Purdue University is now available. This position is ideal for an individual interested in cryo-electron microscopy studies of viruses. The position will involve sample preparation, initial sample assessment, sample freezing, data collection, image analysis and interpretation. A BS with a major in biochemistry or a related area of biology and experience with transmission electron microscopy is considered a minimal requirement. Additional backgrounds in physics and computing along with a willingness to learn and the ability to balance multiple projects would be highly desirable. Employment will entail comprehensive training during the first year and extensive daily interactions with a team of graduate students, post-doctoral scholars and faculty. Our facilities include state of the art 200 and 300kv TEMs. Applicants can find the official posting on Purdue University's website (http://www.adpc.purdue.edu/Personnel/currjobs/lablist.htm). Requests for further information should be sent to Michael Rossmann or Richard Kuhn, Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA. Tel: 765-494-4911; FAX 765-496-1189; e-mail: mgr-at-indiana.bio.purdue.edu or rjkuhn-at-bragg.bio.purdue.edu.
Paul Chipman Research Electron Microscopist Purdue University Structural Virology
I just recei ved an inquiry from a safety engineer asking about the nature of the oil that was used in the high voltage tank of the RCA EML and EMU-3 TEMs. It seem that their university has one of these instruments that has been standing unused for a long time, and which now is to be disposed of, and which he has the task of doing the disposing. His problem is that he doesn't know what the character of the several gallons of oil in the high voltage tank might be, and what the proper disposal procedures are for it.
As I recall the high voltage tanks on some TEMs of that vintage were filled with polychlorinated biphenyl fluids. Then, after these fluids fell into disfavor highly refined hydrocarbon oils were used. However, I don't know what RCA was using at the time (circa 1955).
Any reliable info anyone can offer will be much appreciated. Incidentalyy, if anyone would like to have the instrument for a museum piece (it is now nearly 50 years old and therefore officially an antique) let me know before the disposal process goes too far.
Best regards, Wil Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Dear Wil I would suspect, the oil in the old EM is pretty the same as in the X-ray generators. Probably, regulation for X-ray Roentgen stations in the hospital should be OK for the vintage EMs. Your safe engineer, probably, don't need to know the exact formulation for disposal purpose. If s/he is so curious, s/he may perform mass-spectrum analysis for exact formulation. Another idea comes to me: high voltage transformers in power grill. Sergey
At 02:09 PM 5/5/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Rick Perhaps, you need some more sophisticated technique to see protein incorporated into lipid bi-layer. I would suggest cryo-EM of non-stained samples. I do see a couple problems in your neg-staining setup: you will not see the part, incorporated into the lipid because water solution will not penetrate into lipids, so you may see only extra-membrane portion of the protein. I afraid, plastic film, you are using is too thick for such job, you better use thin carbon film. Neg staining in general deformed the liposome (they becomes flat), so there is no way to do morphometry and so on such samples. Neg-staining is OK just to check liposomes quality (size, homogenesity, concentration etc). Sergey
At 12:13 PM 5/5/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
-----Original Message----- } From: Monson, Frederick C. Sent: Tuesday, May 06, 2003 8:15 AM To: 'Wil Bigelow'
Listers:
I just recei ved an inquiry from a safety engineer asking about the nature of the oil that was used in the high voltage tank of the RCA EML and EMU-3 TEMs. It seem that their university has one of these instruments that has been standing unused for a long time, and which now is to be disposed of, and which he has the task of doing the disposing. His problem is that he doesn't know what the character of the several gallons of oil in the high voltage tank might be, and what the proper disposal procedures are for it.
As I recall the high voltage tanks on some TEMs of that vintage were filled with polychlorinated biphenyl fluids. Then, after these fluids fell into disfavor highly refined hydrocarbon oils were used. However, I don't know what RCA was using at the time (circa 1955).
Any reliable info anyone can offer will be much appreciated. Incidentalyy, if anyone would like to have the instrument for a museum piece (it is now nearly 50 years old and therefore officially an antique) let me know before the disposal process goes too far.
Best regards, Wil Bigelow -- Wilbur C. Bigelow, Prof. Emeritus Materials Sci. & Engr., University of Michigan 3062 Dow Bldg.; 2300 Hayward St. Ann Arbor, MI 48109-2136 e-mail: bigelow-at-umich.edu; Fx:734-763-4788; Ph:734-662-5237
Lista de E-MAILS DE COLOMBIA 255.000 Direcciones Empresariales
Hemos terminado la actualización de direcciones E-mail empresariales de Colombia. 255.000 direcciones Verificadas a Mayo 1 del 2003. Son direciones Empresariales de mas de 35.000 sitios de empresas Colombianas o directamente relacionadas a Colombia Garantizados Se entrega en Archivo con formato Excel® o archivo plano listo para importar a su software de envio de correo. Para compra en Pesos en Colombia. envíe sus datos a: colpe3-at-netscape.net Para compra en US$ con tarjeta de credito, envie sus datos a: coldol3-at-netscape.net Precio Económico. Por menos de lo que usted invierte en un aviso de prensa, obtenga el medio y uselo convenientemente. Disponibles tambien listas de correo E-Mail de Mexico, Chile, Costarica, Guatemala. Para mas información envie sus datos a: otrospais-at-netscape.net Opcional : Software para envío masivo de E-Mail
Rick: To add to Sergey's comments, projections of vitrified liposomes will give you nice bilayer definition but you still may want to consider a gold-conjugated Ab to an exposed part of your protein to confirm its presence. Don
Donald Gantz Dept. Physiology & Biophysics Center Advanced Biomedical Research Boston Univ. School of Medicine Boston, MA 02118 Email: gantz-at-biophysics.bumc.bu.edu Phone: 617-638-4017 (voice mail)
I goofed this morning while updating the Junk Mail filters trying to get rid of the Spammer from Colombia. Sorry gang, but nearly all mail today was rejected.
If you were rejected, please try reposting your message. I hopefully got things fixed and things are back to some semblance of normal operation.
:-(
Nestor Your Friendly Neighborhood (and Ocassionaly Human) SysOp
Disclaimer to all: if this subject has been breached and burned out before - I apologize.
We are currently looking to purchase a system(s) that will meet our sample requirements for both metal deposition carbon evaporation. differences with regards to cost.
What I could use is feedback from users of various systems. I am interested in the reliability, repeatability and deposition consistency. The equipment will be used to } support EMPA and SEM analysis.
Thank you, Evelyn
Scripps Institute of Oceanography University of California, San Diego
I suppose you need to establish, if you haven't already, how the vibration is getting to the TEM. If the vibration is being transmitted through the water line (rather than being transmitted from the chiller through the floor), then isolating the chiller body won't help - you need to dampen the pulses in the water resulting from the rotor vanes in the pump.
Since water doesn't really compress (or expand) under the kind of pressures we're dealing with I'm not sure that putting restrictors on the line would help much, although it will reduce the flow too, which may be bad. Gary Brown suggests any extra length of flexible tubing - another quite elegant way of achieving the same thing is by mounting an ordinary filter housing upside down, minus filter, in line with the output of your chiller. This looks slightly odd but it works well, as the few litres of compressible air trapped in the top of the housing adsorbs much of the pressure fluctuation. A JEOL engineer suggested this inexpensive remedy during the installation of a FE-SEM here recently, seems to work well.
Regards,
Richard
} } On Friday, May 2, 2003, at 04:24 AM, L. D. Marks wrote: } } } We want to reduce/eliminate vibration associated with some } } water cooling lines on our TEM. One suggestion has been to } } mount the water coolers (Haskris) on a platform supported } } by truck tire inner tubes (to prevent ground transmission) } } and put some constrictions in the water line to reduce } } vibration transmission. The first one sounds good; I suspect } } that some sort of in-line filter would be better to remove } } vibrations in the water. } } } } I would be interested in how others have approached this; } } I think it is a fairly general issue. =
Richard Easingwood Otago Centre for Electron Microscopy Department of Anatomy and Structural Biology School of Medical Sciences, University of Otago PO Box 913, Dunedin NEW ZEALAND Telephone: office: 0064 3 479 7301 Facsimile: 0064 3 479 7254 GSM: 0064 21 222 4759 mailto:richard.easingwood-at-stonebow.otago.ac.nz Web site: http://www.otago.ac.nz/anatomy/emunit/ or http://anatomy.otago.ac.nz/ocem/contact.html
If anyone has an old Reichert Ultracut suitable for spares that they would be willing to part with (preferably in the UK), could they kindly contact me off list.
Many thanks.
Bob Phillips
MicroServiS 11 Grafton Close, St. Ives, Cambridgeshire, PE27 3DL UK bob.phillips-at-microservis.co.uk www.microservis.co.uk
Laurence, My experience has been that the majority of the vibration comes from the return line. When you adjust the feed pressure, the system bypasses the excess flow directly to the return (assuming you have a positive displacement pump instead of a centrifugal pump).
A quick check, and sometimes permanent solution, is to remove your return line from the chiller and cap the chiller side. Then provide a direct return to the holding tank, eliminating back-pressure. The mixing will not be as good as the manufacturer's set-up, but there should be a large reduction in the pulses. The by-pass from the pressure regulator will still use their return system.
If you can't find the diaphragm units mentioned earlier, you can build a copper tube system that consists of a tee with horizontal connections to your water lines and a vertical capped section full of air. The tubing should be considerably larger than the inlet and outlet fittings. The main drawback is that you will have to recharge the air from time to time due to it dissolving in the water. The diaphragm eliminates this problem.
Ken Converse owner Quality Images third party SEM service Delta, PA
L. D. Marks wrote:
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I would be grateful if you would post this position advertisement and bring it to the attention of those who might be interested.
Thanks, Susan Belfry, UNB, Fredericton, *********************************** Employment Opportunity
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The University of New Brunswick is seeking an Electron Microscopist - Materials Science Specialist to join the Microscopy and Microanalysis Facility. This is a regional facility providing research services both to the University community and to a wide range of external clients from universities, government and industry in the Maritime provinces. The Microscopy and Microanalysis Facility is currently undergoing a major upgrade to its facilities. The expanded range of instrumentation will include both transmission and scanning electron microscopes, an electron microprobe and a confocal laser scanning microscope. Analytical equipment will include EDS, WDS, and EELS. The successful applicant will join an existing team of three support personnel operating the facility.
Responsibilities: The incumbent will be responsible primarily for the application of imaging and microanalysis to the investigation of materials and duties will include the operation and maintenance of a new 200KV analytical STEM equipped with EDS, EELS, and cryo capabilities.
Requirements: Applicants should have the minimum of a university undergraduate degree in physical science or engineering, plus several years experience working with electron beam instruments, microanalytical equipment, or related instrumentation, preferably with a materials focus. Applicants should also have a sound knowledge of electrical, electronic and vacuum systems, together with experience in troubleshooting and repairing these systems. Specific experience with EFTEM/EELS applications, scientific computer systems and the nature and composition of materials would be a considerable asset. Applicants must also possess good organizational and interpersonal skills and be proficient in verbal and written communications.
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Since y'all are discussing disposing of old oil from a high-voltage tank, I thought someone might have an idea where to purchase such oil for refilling a high voltage tank on an SEM. Someone drained the HV tank before they stored the microscope, thinking the oil was somehow too hazardous to leave in it. I don't know how they disposed of it.
I'm told that the brand and type for this specific instrument (1984 ISI DS-130) is Shell Diala. I found it on the web, but the smallest quantity the distributor sells is a 55 gallon drum for $174 (which is not out of the question, just wasteful if the tank only takes 7-10 gallons). Any ideas on where to get 10 gallons of suitable dielectric oil?
Thanks in advance.
Regards, Andrew T. Werner Chief Metallurgist Shaped Charge Research - Metallurgy Laboratory Schlumberger Reservoir Completions Technology Center 14910 Airline Road, Rosharon, TX 77583-1590 Voice (281) 285-5272 Fax (281) 285-5273
Enclosed is the table of contents for the May issue of Microscopy Today.
I will close the magazine's subscription list on Friday, May 9th for this issue.
MT is free for any one interested in, or connected with, microscopy anywhere in Canada, Mexico, and the USA. MT is also free for Microscopy Society of America members anywhere.
The subscription rate for those not qualifying for free subscriptions has been reduced from $80 or $110 to $35US per year.
Please subscribe via our web page: http://www.microscopy-today.com
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Ron Anderson, Editor Microscopy Today
Cryoelectron Tomography Stephen W. Carmichael, Mayo Clinic Optimizing the Sampling Design of Morphometric Experiments John M. Basgen, University of Minnesota FTIR Hyperspectral Images of Microscopic Droplets of Splattered Blood Scott W.Huffman*, Kara B. Lukasiewicz** and Chris W. Brown*** *NIH, **Mayo Clinic, ***University of Rhode Island Setting White, Black and Gamma on Continuous Tone Grayscale and Color Images using Photoshop Jerry Sedgewick, University of Minnesota How to Recognize and Avoid AFM Image Artifacts Paul West and Natalia Starostina, Pacific Nanotechnology, Inc. Quality in Electron Microscopy Tony Bruton*, Steve Chapman** and Paul Harding*** *E.M. Unit, University of Natal, **Protrain Courses, ***Integrated Systems Management, Nissan Motor Co. Use of a New Imaging Technique to Document Deformations Recorded in the Environmental Scanning Electron Microscope Edward F. O’Neil,* Hamlin M. Jennings,** and Jeffrey J. Thomas** *US Army Corps of Engineers, **Northwestern University On The Limits Of Limits Tobias I. Baskin, Division of Biological Sciences, University of Missouri Examination of Hydrated Bacteria in An Environmental Scanning Electron Microscope (ESEM) Ann Fook Yang and Miloslav Kaláb, Agriculture and Agri-Food Canada Dye is Money Christian Lohr, University of Kaiserslautern, Germany Water Recirculator (Chiller) Maintenance Owen Mills, Michigan Technological Institute EDS Spectral Artifacts / Sum Peaks: A Reminder Steven S. Hurban, Endicott Interconnect Technologies, Inc.
A colleague is looking for a used (or surplus to requirement) field cancelling system for use with an SEM.
Best regards,
Mike Wombwell
**************************************************************************** ***************************** Quorum Technologies Unit 15A, Euro Business Park New Road, Newhaven East Sussex, BN9 0DQ, UK Tel: +44(0)1273 510535 (main switch board) Tel: +44(0)1273 511063 (direct line) Mobile +44(0)7989 426 431 Fax: +44(0)1273 510536 mike.wombwell-at-quorumtech.com http://www.quorumtech.com
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The University of Central Florida is an Equal Opportunity/Affirmative Action Employer As an agency of the State of Florida, UCF makes all search materials, including transcripts, available for public review upon request.
Hi gang - what is your best receipe for cells in culture on a small filter for SEM? This guy is growing astrocytes on one side of filter and on the other side he is growing endothelial cells. He wants to look at blood barriers. Any ideas will be greatly appreciated. Thanks Barb
We have discontinued making single hole Be grids for a variety of reasons. We can do a single hole 3 mm disc with holes ranging from 0.5um to 2 mm in a variety of other materials such as SS, aluminum, tantalum, moly, Pt, tungsten etc. Much depends upon the aspect ratio of the hole dia to thickness. Let me know if we can assist.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "David Henriks" {henriks-at-southbaytech.com} To: "K.N. Bozhilov" {bozhilov-at-mail.ucr.edu} Sent: Thursday, May 08, 2003 12:09 PM
I would like to know supplier of high quality Carbon rod (99.9999% or better). 1/8 inch is fine. Please advise.
Thank you,
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
To: Vendors of used light microscopes. See the following message for details:
} I'm only looking for the basic frame; I'll remove the } optics so they can be busted or missing or whatever. This is } something like what I'm after: } } http://www.geocities.com/nicholl.geo/stuff/d6/aoframe.jpg } } Any scope with an X-Y mechanical stage (the only critical } part) and a basic focus mechanism, then I'm set.
Please reply to Scott Childs {scott-at-rre.net} thanks for your help, Beth
********************************************************************** Beth Richardson EM Lab Coordinator Plant Biology Department University of Georgia Athens, GA 30602-7271
Phone - (706) 542-1790 & FAX - (706) 542-1805
"Between the two evils, I always pick the one I never tried before". Mae West (1893-1980) **********************************************************************
"And it's only the giving that makes you what you are". Wond'ring Aloud, Jethro Tull (Aqualung)
I do believe the smallest quantity you can buy is 55 Gallons - at least in the US. For that reason, I have a significant quantity surplus to my requirements (about 30 gallons, in fact) - but it would probably cost more to ship it to you in Texas than it would for you to but your own 55 gallons! (My oil is actually Exxon Univolt 60, but I think that and Shell Diala are equivalents).
Years ago, you could get this oil in 5 gallon containers, but they leaked (as I can attest). I was told this was the reason they were discontinued.
Tony.
At 08:20 AM 5/7/2003 -0500, Andrew Werner wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
** Anthony J. Garratt-Reed ** Manager, CMSE Shared Experimental Facilities ** MIT Rooms 13-1027 or 13-3090 ** 77 Massachusetts Avenue ** Cambridge, MA 02139-4307 ** USA ** ** Phone: (+) 1-617-253-4622 ** Fax: (+) 1-617-258-6478 **
A number of years ago I was able to dispose of old oil (no PCBs) and get a 5-gallon carboy of new from our local electric company. They also had facilities for disposing of PCB-contining oils. I'm fairly sure that these days I'd have to fill out EPA forms and all, but a few years ago the electric comapany was amenable because 5 gallons was peanuts to them! You might look into official ways to do the same.
Good luck!
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* ****************************************************************************
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (kelidrwil-at-seneca24.net) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, May 8, 2003 at 09:07:15 ---------------------------------------------------------------------------
Email: kelidrwil-at-seneca24.net Name: R.F.Wilson
Organization: Self
Education: Graduate College
Location: Waterloo NY USA
Question: What is the best method for mounting pollen for study with the optical microscope ?
Check with a local power company, they use the oil in high voltage transformers in the powergrid. I've been able to buy small quantities when I needed it for a project.
As a sidenote, a full tankwaggon was stolen here in Umeå a couple of years ago. It was standing outside a powerstation and somebody thought it contained diesel.... I guess a number of cars broke down that winter. :-)
Regards, Göran Axelsson
Anthony J. Garratt-Reed wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I do believe the smallest quantity you can buy is 55 Gallons - at } least in the US. For that reason, I have a significant quantity } surplus to my requirements (about 30 gallons, in fact) - but it would } probably cost more to ship it to you in Texas than it would for you to } but your own 55 gallons! (My oil is actually Exxon Univolt 60, but I } think that and Shell Diala are equivalents). } } Years ago, you could get this oil in 5 gallon containers, but they } leaked (as I can attest). I was told this was the reason they were } discontinued. } } Tony. } } At 08:20 AM 5/7/2003 -0500, Andrew Werner wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Since y'all are discussing disposing of old oil from a high-voltage } } tank, I thought someone might have an idea where to purchase such oil } } for refilling a high voltage tank on an SEM. Someone drained the HV } } tank before they stored the microscope, thinking the oil was somehow } } too hazardous to leave in it. I don't know how they disposed of it. } } } } I'm told that the brand and type for this specific instrument (1984 } } ISI DS-130) is Shell Diala. I found it on the web, but the smallest } } quantity the distributor sells is a 55 gallon drum for $174 (which is } } not out of the question, just wasteful if the tank only takes 7-10 } } gallons). Any ideas on where to get 10 gallons of suitable } } dielectric oil? } } } } Thanks in advance. } } } } Regards, } } Andrew T. Werner } } Chief Metallurgist } } Shaped Charge Research - Metallurgy Laboratory } } Schlumberger Reservoir Completions Technology Center } } 14910 Airline Road, Rosharon, TX 77583-1590 } } Voice (281) 285-5272 Fax (281) 285-5273 } } } } } ** Anthony J. Garratt-Reed } ** Manager, CMSE Shared Experimental Facilities } ** MIT Rooms 13-1027 or 13-3090 } ** 77 Massachusetts Avenue } ** Cambridge, MA 02139-4307 } ** USA } ** } ** Phone: (+) 1-617-253-4622 } ** Fax: (+) 1-617-258-6478 } ** } } }
Here's an article on the subject that might be of use.
http://www.nttworldwide.com/tech2308.htm
Good luck, your problem does not have a simple solution.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Andrew Werner [mailto:werner-at-rosharon.oilfield.slb.com] Sent: Wednesday, May 07, 2003 9:21 AM To: Microscopy Listserver
Since y'all are discussing disposing of old oil from a high-voltage tank, I thought someone might have an idea where to purchase such oil for refilling a high voltage tank on an SEM. Someone drained the HV tank before they stored the microscope, thinking the oil was somehow too hazardous to leave in it. I don't know how they disposed of it.
I'm told that the brand and type for this specific instrument (1984 ISI DS-130) is Shell Diala. I found it on the web, but the smallest quantity the distributor sells is a 55 gallon drum for $174 (which is not out of the question, just wasteful if the tank only takes 7-10 gallons). Any ideas on where to get 10 gallons of suitable dielectric oil?
Thanks in advance.
Regards, Andrew T. Werner Chief Metallurgist Shaped Charge Research - Metallurgy Laboratory Schlumberger Reservoir Completions Technology Center 14910 Airline Road, Rosharon, TX 77583-1590 Voice (281) 285-5272 Fax (281) 285-5273
Jim Darley, at Proscitech (www.proscitech.com.au) has both graphite and carbon 1/8" rods, and ships to anywhere at good prices
I have no connection apart from being a regular satisfied customer.
cheers
rtch
To: Microscopy-at-sparc5.microscopy.com
Howdy Fred,
Thank you (and several others! - I need to be better at responding) for your help and suggestions.
David Sundin at DS Fluids will sell me 5 gallon pails but, since the remaining portion of the drum will require re-processing, cost for 10 gallons will be the same as 55 gallons. It is a viable option though - does not waste the rest.
Others have suggested the local electric utility - a possibility but finding the right person to ask would be a challenge - or donating the excess from a 55 gal drum to the Physics Dept. of the University.
One person referred me to an outfit in Houston that sells him Exxon Univolt, and I have a call in to the sales dept. If this pans out it will be the exact answer to my problem.
Thanks again to all who offered advice. If it is not wasting your time, I will post about how this turns out.
Regards, Andrew
At 03:08 PM 5/9/2003 -0400, Monson, Frederick C. wrote:
} Afternoon Andrew, } It appears that you are within hailing distance of an expert in the } area of dielectric oil replacements, etc. } } These folks certainly appear to concentrate on your problem. Perhaps they } would provide a 25Gal lot for a fellow Texan. } } http://www.dsifluids.com/dielectric_fluids_home.htm } } http://www.dsifluids.com/beta.html } } Here's an article on the subject that might be of use. } } http://www.nttworldwide.com/tech2308.htm } } Good luck, your problem does not have a simple solution. } } Cheers, } } Fred Monson } } Frederick C. Monson, PhD } Center for Advanced Scientific Imaging } Mail Drop: Geology } West Chester University } West Chester, PA, 19383 } http://darwin.wcupa.edu/casi/ } Phone/FAX: 610-738-0437
Email: daniel.smith-at-liquidminerals.com Name: Daniel Smith
Organization: Liquid Minerals Group, Inc.
Education: Graduate College
Location: Houston, TX USA
Question: I am interested in measuring particle size of metal oxide particles suspended in a process oil. The particles are on the order of 1 - 300 nanometers. I have used indirect methods (DLS, etc.), but I would really like to take photos of the particles and determine particle size, crystal structure, agglomeration and other properties.
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Hiromi Konishi wrote: ================================================================== I would like to know supplier of high quality Carbon rod (99.9999% or better ). 1/8 inch is fine. Please advise. ================================================================== The purity of carbon and/or graphite rods is normally discussed in terms of so many ppm ash left over after ashing in a high tempeature muffle furnace. I understand that there is some amount of "controvery" with regard to the **exact** meaning of these terms, but generally speaking, if the carbon (or graphite) material exhibits less than 10 ppm ash, it is described as being "spectrographic" purity, and if less than 6 ppm ash, then "spectroscopic" purity.
As you might imagine, there is a difference in the cost to reduce the ash content from 10 down to 6 ppm. SPI Supplies offers both spectroscopically pure carbon and also graphite rods in "spectroscopic" purity as standard products, in 1/8" and other diameters. Several of our major competitors offer similar purity, at least in terms of graphite rods, so our product is not unique in that respect.
However, it is not clear if you really wanted (literally) **carbon** or if you wanted graphite rods (but like many of us tend to call them "carbon"). More information about our carbon and graphite rod products and their purity can be found on URL www.2spi.com\catalog\spec_prep\carbon-rods.shtml
For vacuum evaporation work, most workers seem to prefer graphite to carbon rods.
Disclaimer: SPI Supplies offers both carbon as well as grahite rods in high purity (less than 6 ppm ash).
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 ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id TAA00806 for dist-Microscopy; Sun, 11 May 2003 19:01:16 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id TAA00801 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Sun, 11 May 2003 19:00:45 -0500 (CDT) Received: from mailhost2.auckland.ac.nz (mailhost2.auckland.ac.nz [130.216.191.4]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id TAA00789 for {Microscopy-at-sparc5.microscopy.com} ; Sun, 11 May 2003 19:00:32 -0500 (CDT) Received: from glgnov2.auckland.ac.nz (glgnov2.auckland.ac.nz [130.216.59.90]) by mailhost2.auckland.ac.nz (8.12.9/8.12.9/8.12.9-ua) with ESMTP id h4BNracX021457; Mon, 12 May 2003 11:53:36 +1200 (NZST) Received: from GLGNOV2/SpoolDir by glgnov2.auckland.ac.nz (Mercury 1.48); 12 May 03 11:54:51 +1200 Received: from SpoolDir by GLGNOV2 (Mercury 1.48); 12 May 03 11:54:50 +1200 MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
This may well be so, but my old Edwards 306 and I prefer 'carbon', as the graphite ones need appreciably higher temperature to evaporate it, and the rod holders get realy hot.
Give me 'carbon' any day
cheers
rtch
To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Hiromi Konishi wrote: ================================================================== I would like to know supplier of high quality Carbon rod (99.9999% or better ). 1/8 inch is fine. Please advise. ================================================================== The purity of carbon and/or graphite rods is normally discussed in terms of so many ppm ash left over after ashing in a high tempeature muffle furnace. I understand that there is some amount of "controvery" with regard to the **exact** meaning of these terms, but generally speaking, if the carbon (or graphite) material exhibits less than 10 ppm ash, it is described as being "spectrographic" purity, and if less than 6 ppm ash, then "spectroscopic" purity.
As you might imagine, there is a difference in the cost to reduce the ash content from 10 down to 6 ppm. SPI Supplies offers both spectroscopically pure carbon and also graphite rods in "spectroscopic" purity as standard products, in 1/8" and other diameters. Several of our major competitors offer similar purity, at least in terms of graphite rods, so our product is not unique in that respect.
However, it is not clear if you really wanted (literally) **carbon** or if you wanted graphite rods (but like many of us tend to call them "carbon"). More information about our carbon and graphite rod products and their purity can be found on URL www.2spi.com\catalog\spec_prep\carbon-rods.shtml
For vacuum evaporation work, most workers seem to prefer graphite to carbon rods.
Disclaimer: SPI Supplies offers both carbon as well as grahite rods in high purity (less than 6 ppm ash).
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 ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id KAA03944 for dist-Microscopy; Mon, 12 May 2003 10:11:36 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id KAA03939 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Mon, 12 May 2003 10:11:05 -0500 (CDT) Received: from tumor.soft-imaging.com ([67.104.115.34]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id KAA03931 for {Microscopy-at-sparc5.Microscopy.Com} ; Mon, 12 May 2003 10:10:53 -0500 (CDT) Received: from lakewood.soft-imaging.com (lakewood.soft-imaging.com [192.168.5.225]) by tumor.soft-imaging.com (8.10.2/8.10.2/SuSE Linux 8.10.0-0.3) with ESMTP id h4CF4lc19564 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 12 May 2003 09:04:47 -0600 Received: by hq-dc2.soft-imaging.com with Internet Mail Service (5.5.2656.59) id {J6F4H6QZ} ; Mon, 12 May 2003 09:04:48 -0600 Message-ID: {6127CE87B9BDD511B59D0001028A497D478685-at-hq-dc2.soft-imaging.com}
Daniel,
I think you need to be more specific. Are you looking for help regarding SEM or TEM, sample preparation, observation methods, image acquisition, further image processing, etc?
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: daniel.smith-at-liquidminerals.com [mailto:daniel.smith-at-liquidminerals.com] Sent: Friday, May 09, 2003 5:28 PM To: MicroscopyListserver
Email: daniel.smith-at-liquidminerals.com Name: Daniel Smith
Organization: Liquid Minerals Group, Inc.
Education: Graduate College
Location: Houston, TX USA
Question: I am interested in measuring particle size of metal oxide particles suspended in a process oil. The particles are on the order of 1 - 300 nanometers. I have used indirect methods (DLS, etc.), but I would really like to take photos of the particles and determine particle size, crystal structure, agglomeration and other properties.
A colleague would like to purchase a camera for use in phase contrast and fluorescence microscopy of bacteria. For some of his specimens, he expects the fluorescence may be weak.
He would like a camera with a non-proprietary (firewire) interface, and any included software should be available for the Macintosh platform.
Among the cameras he is interested in are the Qimaging Micropublisher and Qimaging Micropublisher 5.0, and the Optronics Magnafire and Microfire. He would like to have feedback from anyone who has experience with one or more of these cameras, or anyone who has evaluated one of them but has chosen another camera.
Thank you,
Ed King --
Edward J. King Department of Biology University of Utah 257 South 1400 East Salt Lake City, UT 84112
Hi, we have sprung a water leak from our 3 year old TEM. We are using mains 'tap water' to cool as this was deemed (at the time) better than installing a chiller system. Has anyone any experience/advice they can give me? Is this a common occurrence?
Gillian Brown Histology Section, Asthma Biology RI CEDD, GlaxoSmithKline UK gillian.2.brown-at-gsk.com
Do you mean carbon thread? I'm trying to figure out what is better for our samples, graphite rods or carbon thread? Any information will be appreciated. Thank You, Evelyn
At 11:54 AM 5/12/2003 +1200, Ritchie Sims wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A colleague of mine needs to cut frozen sections of adult Drosophila heads for a combination immunohistochemistry/in situ hybridization study. Any suggestions?
Thank you very much,
Joel
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 6646 http://astro.temple.edu/~jbs
The College of Physical & Mathematical Sciences at Brigham Young University (BYU) invites applications for a technical administrative staff position in support of the transmission electron microscopy laboratory. The position will be administered in the Department of Physics & Astronomy.
Facility:
The laboratory is located in a new state-of-the-art facility that will house both a 200KV ultra-high-resolution (S)TEM and a 300KV TEMs, with multiple specimen preparation support labs. Analytical capabilities include EDS and EELS.
Requirements:
An M.S. degree or comparable experience, (Ph.D. degree preferred) in physics, materials science, or a related field is required, plus at least 2 years experience in operating and maintaining transmission electron microscopes for materials applications. The successful candidate must have the ability to train users in both sample preparation and TEM use, to interact effectively and collaborate with users from advanced undergraduates through faculty from a variety of disciplines, and to maintain the instruments and interact with manufacturersservice engineers, as appropriate. BYU is sponsored by The Church of Jesus Christ of Latter-day Saints (LDS, Mormon) and is an equal employment opportunity/affirmative action employer. Strong preference is given to LDS applicants. Employees are required to abide by standards consistent with LDS values.
Application:
Contact E. Daniel Johnson, N-181 ESC, BYU, Provo, UT 84602. Applications will be accepted until July 1, 2003 or until a successful candidate is identified, whichever is later. This is a permanent, full-time, budgeted position.
On Friday, May 9, 2003, at 04:28 PM, by way of MicroscopyListServer wrote:
} Question: I am interested in measuring particle size of metal oxide } particles suspended in a process oil. The particles are on the order } of 1 - 300 nanometers. I have used indirect methods (DLS, etc.), but } I would really like to take photos of the particles and determine } particle size, crystal structure, agglomeration and other properties. } } Any help would be appreciated. } Dear Daniel, You can use TEM to image the particles, but you'll have to get rid of the oil. If it is a volatile oil, things may be as simple as putting a microliter or so on a formvar-carbon grid, evaporating the oil, and inserting the grid into the scope; however, if this can't be done, you may lose some of the particles in the separation process--e.g., the smaller ones could be trapped on a filter membrane. If the oil is soluble in something like acetone or chloroform, you might be able to add about 1ml of solvent, centrifuge the particles, resuspend them, and place an aliquot on a formvar-carbon grid. (Here, carbon is essential, since the formvar will dissolve in the solvent.) Particle size and structure should be measurable, with the caveat that, if you lose particles, the size distribution will likely be altered. Agglomeration could easily be lost, since, even if you have a volatile oil, the particles can aggregate as the oil evaporates. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Senior EM Technician position available in the Department of Pathology at George Washington University Medical Center.
Must have experience in all phases of TEM: fixation, embedding, thick & thin sectioning, scoping and scope maintenance(filament changing and alignment, mostly).
Experience with clinical samples a plus, but not neccessary.
This is a service lab mostly and handles primarily clinical samples though we do get some research samples.
Interested?
Contact by e-mail at patpxs-at-gwumc.edu
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Charles Interestingly and without any "scientific" explanation, I do find that "carbon (not graphite) rods" works better to me. Somehow the properties of the evaporated film is different. To me, films from "carbon" (not graphite) is "better"... Rational (not necessary scientific) explanation may be that we have deal with "sublimation" of carbon/graphite. During this process we generate a cloud of particles (clusters), which being condensed on the surface formed the film. We have to understand that thermal evaporation of carbon creates actually particles from a few to tens atoms in size, it's not mono-atomic cloud. Therefore, the film quality will dependent from the composition of cloud. Personally, I prefer "Electron Gun" carbon evaporation: the films are much more stable and uniform. Have a good day. Sergey
At 11:17 PM 5/10/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
I'm shopping around for a replacement for the inkjet printer I've been using to output my SEM images. I have been using an HP 970 very happily, but it is getting worn and cranky about feeding paper consistently. I have tried cleaning the feed rollers (have lots of green ink on them by this time), but with limited success.
My reason for writing is that the SEM data system (Hitachi S3000N) uses Windows 95 (haven't gotten funding for the overhaul to allow running Windows 2000) and it does not have USB ports available. In principle USB might work under late Windows 95, but I haven't gotten that going. So I am limited to parallel port or Ethernet connections to the computer. I also hope to get a system with more than a small amount of memory built in to help speed up the return of control to the microscope while the images print out.
Any recommendations for a good printer to use based on these limitations?
I have some choices, but none seem 100% satisfactory. I'd be very happy with an HP 990, but those are off the market and the HP 995 only does USB and wireless. A number of possibilities are Windows 98 and up only. Earlier I tried an HP CP1160, but I wasn't happy with the image tones despite aggressive adjustments to the printing and it also seemed to require frequent ink changes (small cartridges only available). Image quality was ok although it was slower than the 970. So I'm wondering about other choices (Epson, Canon) before buying a unit blind and being stuck with it like I am with the 1160.
Richard Shalvoy Arch Chemicals, Inc. 350 Knotter Drive Cheshire, CT 06410 (203) 271-4394 rbshalvoy-at-archchemicals.com
Sorry for any double posting. I'm just learning how to use this listserv.
A colleague of mine needs to cut frozen sections of adult Drosophila heads for a combination immunohistochemistry/in situ hybridization study. Any suggestions?
Thank you very much,
Joel
Joel B. Sheffield, Ph.D. Biology Department, Temple University 1900 North 12th Street Philadelphia, PA 19122 jbs-at-temple.edu (215) 204 8839, fax (215) 204 6646 http://astro.temple.edu/~jbs
We are looking for an experienced EM tech. for a busy Clinical service lab. Details are online at:http://employ.uchc.edu/ It is listed under Anatomic Pathology Search code 2003-161.
Where in the cooling system did the leak occur? I assume it wasn't a coupling or someplace obvious. If it wasn't, I would look into two possible causes: 1. You have electrolysis from improper grounding between the cooling system and/or your TEM. 2. Turbulence from air in the water lines are eroding the copper cooling lines.
We have had the latter happen to us this past year. Nothing spoils a morning more than walking into your lab and finding water all over the floor. Especially if the pumps are running and there is water between you and them, along with a few power supplies in the high kV range.
Good luck, I hope this helps.
Bill
Bill Carmichael EM Faculty Madison Area Technical College (MATC) 3550 Anderson St. Madison, Wisconsin 53562 http://www.matcmadison.edu/electronmicros wcarmichael-at-matcmadison.edu
Ladd Research produces hundreds of carbon substrates daily and this is what we've found concerning carbon and graphite.
Carbon 1. Requires less current 2. burns cooler 3. doesn't chunk as much
Graphite 1. Sparks more, especially with a poor vacuum. 2. Messier to handle 3. Easier to cut and point
Since we cut and point a great deal of rods our personnel prefer to use graphite. The resulting substrates are excellent with either product. Since we also manufacture vacuum deposition systems and high volume rod sharpeners we can handle graphite easier. Bottom line is that we feel it is customer's preference, based on the available equipment and volume.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, May 12, 2003 2:58 PM
To whom it may concern:
Does anyone know of an independent service contractor for JEOL TEM's that take care of the Los Angeles area. Our current contractor will soon be retiring.
Thank You,
Michael Pidgeon University of Southern California Dept. of Cell & Neurobiology
We have had a rash of cooling leaks in our facility in copper lines running to and from our refrigerated water recirculators and in the brass plumbing connectors. The pipes and fittings are being eaten out from the inside, resulting in pinhole leaks at unpredictable times. Fortunately, the leaks usually start small and so far we've caught them before the deluge causes major damage.
There have been two causes for these leaks. One is that, until recently, I was unaware of the presence of a shut-off valve inside our water chillers, which turns the cooling water to the chiller on and off as the compressor turns on and off. The water cooling the coils should only be running when the compressor is delivering a heat load to dissipate. (The water from the tank to the scope runs constantly, of course.) These shut-off valves wear out routinely every couple of years and should be checked once a year, at least, I now know. If they wear out, the water runs constantly and can cause major internal "etching" of the metal cooling lines. Also, one of our chillers had the flow rate to the coils set WAY too high, with the same effect. Moral: have a refrigeration person check your shut-off valves and flow rates.
The other reason is that some water supplies are simply more corrosive than others (our refrigeration guy used the term "hungry" water) and will eat your copper lines and metal fittings from the inside out over varying periods of time, regardless of flow rates. Have your water checked for hardness and other water voodoo by some qualified person with a water voodoo test kit. Regular copper lines can be replaced by hardened copper (I think it's called K-copper) or stainless steel lines, but there is an expense factor involved.
So far, our main leak detectors have been our squeaky shoes on wet floors, rather than soaked and electrocuted microscopes. We now watch our lines carefully. One consistent leak trait is that they occur over weekends, so we suspect the Leak God does its thing at 5:15 p.m. on a Friday afternoon, just as I'm ordering a Katy Trail Pale Ale down at the old Flatbranch Brewpub and Grill (no financial interest, just a satisfied customer).
Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Bill Carmichael [mailto:wcarmichael-at-charter.net] Sent: Tuesday, May 13, 2003 9:54 AM To: Microscopy-at-sparc5.microscopy.com
Gillian,
Where in the cooling system did the leak occur? I assume it wasn't a coupling or someplace obvious. If it wasn't, I would look into two possible causes: 1. You have electrolysis from improper grounding between the cooling system and/or your TEM. 2. Turbulence from air in the water lines are eroding the copper cooling lines.
We have had the latter happen to us this past year. Nothing spoils a morning more than walking into your lab and finding water all over the floor. Especially if the pumps are running and there is water between you and them, along with a few power supplies in the high kV range.
Good luck, I hope this helps.
Bill
Bill Carmichael EM Faculty Madison Area Technical College (MATC) 3550 Anderson St. Madison, Wisconsin 53562 http://www.matcmadison.edu/electronmicros wcarmichael-at-matcmadison.edu
Does anyone know of an independent service contractor for JEOL TEM's that take care of the Los Angeles area. Our current contractor will be retiring very soon.
Thank You,
Michael Pidgeon University of Southern California Dept. of Cell & Neurobiology
We too have had these internal leaks in copper cooling lines. Thousands of dollars of damage has been done and many of my hours spent making repairs.
We tracked the source to impingement corrosion, particulate laden water moved through small diameter tubing at high speed. We have (no, had) street water cooled condensers on our refrigeration chillers. As Randy said, without careful monitoring and adjustment, the valves controlling the required amount of street water necessary to remove the heat load stay open all the time allowing tremendous amounts of water to flow through that circuit. At high pressure (our street water is 85 psi) and with some micron size particles in it and it will eat copper tubing at any bends in the tubing. Oh, and yes, always on weekends.
We've also seen the problem on street water cooled diffusion pumps, where the coiled copper tubing soldered to the side wall of the diff stack will perforate. Yes, this tubing can be replaced in the field in a few hours with common plumbing tools.
So the result is we no longer have refrigerant cooled chillers. With Haskris' help, I converted all to water-water cooled systems with the new "braze-pak" style heat exchangers that are resistant to this corrosion. No problems since.
RE: Water. I pulled this bit of info from this list recently posted by Andrei Shchukarev who wrote that Neslab recommends water for long term usage in the 1 to 3 megohn-cm (compensated to 25oC) range.
Speaking of Neslab, they have a nice detail on their big chillers, a level switch that shuts the system down if the tank level drops below the switch setting. With that at least you won't pump all of reservoir onto floor. Easily added to any chiller.
I have no financial interest in either company mentioned in this email.
Owen Owen P. Mills Electron Optics Engineer 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
At 03:55 PM 5/13/2003 -0500, Tindall, Randy D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear Richard, I have been very happy with a couple of Epson inkjet printers I have. They seem to have a good life and never give me any trouble. They are expensive in ink cartridges and the right paper, but cheap to buy initially. I don't think you can put much memory in them. They just are slow. Better to buy several to speed things up. The last time I bought one (one year ago) they could use parallel or USB connection. You can also buy USB cards for your WIN95 computer that should have software. I have also used a Datalink parallel to Ethernet connector to put my laser printer on the network. Using the network, I have five printers in my lab I can print to. I use the 1200 dpi laser printer for quick SEM images and the inkjets for x-ray maps and glossy SEM photos. Good luck. I have a S3000N, too.. Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Shalvoy, Richard B **CHES" {RBShalvoy-at-archchemicals.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, May 12, 2003 1:04 PM
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (s54andrews-at-stanford.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 13, 2003 at 19:01:58 ---------------------------------------------------------------------------
Email: s54andrews-at-stanford.edu Name: Scott Andrews
Organization: Stanford University
Education: Graduate College
Location: Stanford, CA
Question: Hello,
I am trying to find out the proper configuration for aligning a condenser in a microscope. Unlike all information I can find, it is a reflective setup, i.e. the light source passes through the objective on its way to the sample. To make things more complex, in the middle of the aperture kit, there is a diffusing glass plate. This can be removed, but it also removes all apertures (aperture stop and field stop) and a intermediate lens. I am unable to tell what the intermediate lens does. It lies between the aperture stop and field stop. With the diffusive glass plate in place, I cannot image the arc poles on the lamp. The light source is a mercury arc lamp with vertical and horizontal bulb alignment knobs. There is a curved, reflective mirror behind the lamp with tilt and vertical position knobs. There is a final knob that moves the mirror along the optical axis. Can you tell me how to align these five knobs so that I achieve Koehler illumination?
Hi My colleague is looking for a universal stage for optical microscope. Petorologists used it to measure optical properties of minerals probably ~30 years ago. You can rotate thin section three dimensionally using the Universal Stage. Nowadays Leica, Zeiss, Nikon, and Olympus do not produce such stage, so I am looking for a used one. You can see the stage: http://www.gemmary.com/insts/forum/messages/8797.html
If you are selling a universal stage or you know a vender for selling such stage, please advise.
hkonishi-at-unm.edu
Hiromi Konishi, Ph.D. Dept. of earth and Planetary Sciences, The University of New Mexico
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
Charles Supper Company 15 Tech Circle - Natick, MA 01760 Tel: (508) 655-4610 Fax: (508) 655-3913 Toll Free: (800) 323-9645 http://www.charles-supper.com/
Thank you, Bob Voigt (bobvoigt-at-restechimage.com) Resolution Technology, Inc. (www.restechimage.com) Phone (614)921-0045 Fax (614)921-0046
-----Original Message----- } From: Hiromi Konishi [mailto:hkonishi-at-unm.edu] Sent: Wednesday, May 14, 2003 3:07 PM To: Microscopy-at-sparc5.microscopy.com
Hi My colleague is looking for a universal stage for optical microscope. Petorologists used it to measure optical properties of minerals probably ~30 years ago. You can rotate thin section three dimensionally using the Universal Stage. Nowadays Leica, Zeiss, Nikon, and Olympus do not produce such stage, so I am looking for a used one. You can see the stage: http://www.gemmary.com/insts/forum/messages/8797.html
If you are selling a universal stage or you know a vender for selling such stage, please advise.
hkonishi-at-unm.edu
Hiromi Konishi, Ph.D. Dept. of earth and Planetary Sciences, The University of New Mexico
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
The following was sent to the MSA Business Office. We are forwarding it to the microscopy listserver in the hope that the originator can find some help.
Please, DO NOT reply to me. Send your responses to the requestor directly, axelschlitt-at-gmx.net, as he apparently does not subscribe to the microscopy listserver. COPY the listserver if you want.
----- Original Message ----- } From: "Axel Schlitt" {axelschlitt-at-gmx.net} To: {BusinessOffice-at-sparc5.microscopy.com} Sent: Tuesday, May 13, 2003 3:16 PM
Hello Becky,
Have you tried Duke Scientific? Their product line may not include exactly what you need (2mm spheres are glass), but they may have something I missed or know where to send you.
Regards, Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Becky Holdford [mailto:r-holdford-at-ti.com] Sent: Thursday, May 15, 2003 3:09 PM To: Microscopy ListServer
All: one of my engineers needs a source for polystyrene beads approximately 2-3 millimeters in diameter. Any suggestions?
-- ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Becky Holdford (r-holdford-at-ti.com) 972-995-2360 972-648-8743 (pager) SC Packaging FA Development Texas Instruments, Inc. Dallas, TX ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
We use the Leica Ultrastainers in our EM Facility. Recently we've experienced problems with a nasty precipitate on the sections.
We contacted our Leica Representative and their response was as follows:
" We have a hold on all stain. there is an issue with exporting and new EU country laws.Plus leaking bags. We have an interim solution. We can supply a glass bottle and special bracket for you to mount to the stainer. You can then make up your own stain. We hope top have the issues resolved by the middle to end of the summer."
The lot #s in particular are: 16 Oct 2004 and 20 Juni 2004. We checked some older lots from 2003, and they do not have this problem.
By sending this alert, we are hoping to save you the headache we've just gone through trying to figure out what went wrong.
It is unfortunate that Leica didn't issue a recall on these lots. It would have saved us a great deal of trouble.
I have been working on Ovary samples of fishes,but never been sucessful. The infiltration always seem to be incomplete resulting in the specimen becoming powdery on sectioning. I have also faced the same problem while doing Histology of the same and so also is a friend of mine working on amphibian ovaries.
I would be grateful if any of you can give a solution to this problem of mine.
Joston Paul (Jos)Sophisticated Analytical Instrumetation Facality Shillong, Meghalaya.India
__________________________________________________ Yahoo! Plus For a better Internet experience http://www.yahoo.co.uk/btoffer
I am looking for a method for fixing and embedding red blood cells for TEM so that the normal shape and architecture of the cell is preserved. Using my normal TEM fix and embedding, the cells end up all different shapes. Also, does any one have a protocol for freeze substitution embedding of RBC's? I am interested in looking at membranes and cell cytoskeleton.
Thank you David ____________________
David Elliott
Yale University School of Medicine 810 LCI 333 Cedar Street New Haven, CT 06520-8022
This in reference to my previous problem on the TEM sample preparation of Fish Ovary The step-by-step procedure followed are: Primary fixation in cacodylate-buffered 2.5%Glutaraldehyde;washed in 1M Cac buffer; post-fixed in 1M OsO4 and washed in 1M cac-buffer; dehydration was carried out in grades of acetone; cleared in Propylene Oxide; infiltrated in mixtures of Propylene oxide and Spurr embedding medium;Embedding was done in the pure embedding medium and then left for polymerisation. Apart from the above steps I have tried to manipulate the timings, such as fixation, infiltration, etc but to no avail. I would always get powdery sections of the ova. SEE IF YOU CAN HELP ME OUT.
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Does anybody have a Tektronix Phaser IISDX color printer that is still in use ? I need to have some information about the availability of its parts and service.
Thank you!
Pat Sadhukhan Bridgestone/Firestone Research, LLC 330-379-7518
I seem recall from way back in my biological EM days that we fixed RBC's in glutaraldehyde prior to spinning down into a pellet. The alternative to centrifugation may be to allow the fixed RBC's to slowly settle out of the plasma and form a layer that one may be able to dehydrate and embed. I just don't remember if I tried the settling approach or not. Good luck
Cheers,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
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I am looking for a method for fixing and embedding red blood cells for TEM so that the normal shape and architecture of the cell is preserved. Using my normal TEM fix and embedding, the cells end up all different shapes. Also, does any one have a protocol for freeze substitution embedding of RBC's? I am interested in looking at membranes and cell cytoskeleton.
Thank you David ____________________
David Elliott
Yale University School of Medicine 810 LCI 333 Cedar Street New Haven, CT 06520-8022
David- WE fix the cells first in glutaraldehyde and then pellet it down and treat pellet as a bulk and proceed for further processing. Cells do appear in diffeent shapes depending on their orientation in cutting plane. The membranes are well fixed and can be further improved by using tannic acid which imprves visualzation of both membrane and cytoskeleton. Good luck. Shashi Singh CCMB Hyderabad INDIA
I am looking for a method for fixing and embedding red blood cells for TEM so that the normal shape and architecture of the cell is preserved. Using my normal TEM fix and embedding, the cells end up all different shapes. Also, does any one have a protocol for freeze substitution embedding of RBC's? I am interested in looking at membranes and cell cytoskeleton.
Thank you David ____________________
David Elliott
Yale University School of Medicine 810 LCI 333 Cedar Street New Haven, CT 06520-8022
Tel: (203) 785-7573 Fax: (203) 785-3864
__________________________________ Do you Yahoo!? The New Yahoo! Search - Faster. Easier. Bingo. http://search.yahoo.com
Dear Josten, There is problem with ovaries because of yolk, Did you try methacrylates or spurr resin. Give longer infilteration times with diluted grades. Like for spurr start with 1:6(Resin:alcohal/acetone)followed by 1:2, 1:1 and back to 6:1 before going to pure resin. Shashi SIngh CCMB Hyderabad INDIA
I have been working on Ovary samples of fishes,but never been sucessful. The infiltration always seem to be incomplete resulting in the specimen becoming powdery on sectioning. I have also faced the same problem while doing Histology of the same and so also is a friend of mine working on amphibian ovaries.
I would be grateful if any of you can give a solution to this problem of mine.
Joston Paul (Jos)Sophisticated Analytical Instrumetation Facality Shillong, Meghalaya.India
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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PRINCIPLES OF FLUORESCENCE TECHNIQUES COURSE LAMBS, department of Physics, University of Geno, June 11-13, 2003 The use and application of fluorescence techniques is increasing daily both in the academia and in industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence instrumentation and techniques, as well as for researchers and industrial scientists who intend to deepen their knowledge of fluorescence techniques. The theoretical lectures delivered by key scientists in the field are complemented by the direct utilization of steady state and lifetime fluorescence instrumentation provided by leading companies. It is recommended that participants have at least a bachelor's degree in the life science, physical sciences or engineering before attending this course. Topics addressed in this course include:
Basic Definitions and Principles of Fluorescence Fluorescence Polarization Time-resolved Fluorescence Instrumentation Data Manipulation and Data Analysis Fiber Optic Sensors Confocal Fluorescence Microscopy Lifetime Imaging Fluorescence Correlation Spectroscopy
The Principles of Fluorescence Techniques course will be held at: Laboratory for Advanced Microscopy, Bioimaging and Spectroscopy Department of Physics, University of Genoa via Dodecaneso, 33 16146 Genova, Italy Details: www.lambs.it; diaspro-at-fisica.unige.it The number of participants to the Principles of Fluorescence Techniques Course is limited to 40 people. PLEASE RESERVE YOUR PARTICIPATION AS SOON AS POSSIBLE SENDING AN E-MAIL TO: coordinator-at-fluorescence-foundation.org. Details : www.fluorescence-foundation.org
------------------------------------------------------------------------ DO NOT MISS PICTURES and NEWS on www.focusonmicroscopy.org ------------------------------------------------------------------------ - Alberto Diaspro Department of Physics, University of Genoa Via Dodecaneso 33, 16146 Genoa Italy voice +390103536426/480, facsimile +39010314218 visit http://www.lambs.it visit http://www.societaitalianalaserchirurgia.it ------------------------------------------------------------------------ - ALLA RISCOSSA! ALLA RISCOSSA! ALLA RISCOSSA! ------------------------------------------------------------------------ -
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (michael_burnham-at-ceo.cudenver.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 20, 2003 at 09:14:34 ---------------------------------------------------------------------------
Email: michael_burnham-at-ceo.cudenver.edu Name: Michael Burnham
Organization: Kent Denver School
Education: 9-12th Grade High School
Location: Denver, Colorado
Question: My questions about an older Zeiss EM-9 TEM that I've been working on for use in our High School science program. It's now fully functional and working well. However, my most important concern is to make sure that no X-rays are being emitted from around the gun. I had assumed that a Geiger tube would detect the presence of X-rays although I'm not sure that this is true. I've tried this method with negative results, yet I do want to make 100% certain before I have students learning to use this EM.
Can you please advise??
Thank you greatly,
Michael Burnham Kent Denver School Chair Science Dept. 4000 E. Quincy Ave Englewood, CO 80110
I am planning experiments to use a polyclonal antibody against hemagglutinin (Santa Cruz Biotechnology) to localize the HA epitope at the EM level, using a post-embedding protocol on transgenic nematodes expressing an HA-tagged protein. I am uncertain how strongly I can fix the HA epitope and still get specific binding on thin sections.
Does anyone have experience with aldehyde fixations for HA-tagged proteins, either for immunofluorescence or for immunoEM? Experience with any species might be helpful in guessing how the epitope will behave. Will the HA tag survive a fixation with buffered 2.5% glutaraldehyde?
I'm planning a microwave fixation in aldehydes, dehydration through methanol, and embedding into LR Gold.
Thanks in advance for any advice you may have.
Best regards,
Dave -- David H. Hall, Ph.D. Center for C. elegans Anatomy Department of Neuroscience Albert Einstein College of Medicine 1410 Pelham Parkway Bronx, NY 10461
David, Gluataraldehyde is not at all good for immunostudies. Fix your specimens briefly in paraformaldehyde 4% and follow it up for dehydration and LR gold embedding as usual. Set the conditions (fixative, blocking etc)for LM before and then proceed for Immunogold. Shashi
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Dear Colleagues, we are searching for an antibody recognizing the active form of Caspase 3 (mouse) to identify apoptotic cells in cryo sections which were formaldehyde-prefixed. Any experiences, tricks, tips and recommendations are welcome.
Thanks in advance
Daniel
-- (-)-(-) --------------------------- \"/ --- Daniel Eberhard =V= Developmental Biology & Molecular Pathology University of Bielefeld D 33501 Bielefeld/Germany
I am Dr Luisa Pimentel Estrada, the wife of Joseph Ejercito Estrada,former president of Philippines. I have children with my husband Jose, Jacqueline and Jude.Two sons and a daugther.This mail may be coming to you as a surprise or an article but it is very real.I gave the mail to my daugther Jude to send the mail to any contacts she sees and may be a God fearing person will listen to our plight.I will want you as the receiver to read through it and think very well if you can help or render us any assistance. My husband Joseph Ejercito estrada was elected as the 13th President of the Philippines in May 1998 by the people of Philippines due to his popularity in the film industry made him to win the largest popularity in the history of election in Philippines.He has attain the position of Senate in 1987,then vice-president in May 1992 and later become the president 1998.My husband became mayor of his hometown, San Juan in 1969 but it was 1972 that he had a string of public successes. My husband was named one of the ten Outstanding Young Men in Public Administration. He was also named Most Outstanding Mayor and Foremost Nationalist and Most Outstanding Metro Manila Mayor. My husband is recently accused of illegal acquire some four Billion Peso ($80M) during his 31 months in office as President backed up by an uprising of mass Demonstrators and Senate Traitors. They also said that he has skimmed off tobacco excise Taxes benefitting from government business deals.Most of them benefitted from my husband's generousity when he was in office. But they just turned around to be the ones to impeach him. I have tried every possible means to get him out of Detention without success. The Despotic forces in power appear bent in deriding him, rubbishing his achievements while freezing all his known Bank accounts.He has been accused of economic plunder carrying the maximum penalty of death. To the worst of it all,all other wives of my husband especailly Guia Gomez and some of his children born outside wedlock are testifying against us.In conjunction with the PCGG set up by the recent President Arroyo Macapagal Gloria. These are some of the allegations file before my husband in the impeachment trial; 1. Gov. Luis Singson, a longtime friend of my husband, said he provided the my husband with more than $8 million in payoffs from illegal gambling and $2.7 million from tobacco taxes. 2. Witnesses testify one of an account in the Philippines third largest bank held millions of dollars in bribes collected by my husband. Equitable PCI Bank President George Go resigns. The banks senior vice president, Clarissa Ocampo, said she saw my husband sign a false name to documents withdrawing $10 million from a secret personal account. 3. On Dec 31 Five synchronized bomb attacks kill 22 people and injure more than 120 in Manila, days before the trial is to return from holiday recess. Police accuse Muslim rebels but many fear the bombs may be linked to the trial. 4. That my husband received about $8.5 million in pay-offs from illegal gambling operators. 5. That my husband participated in a real estate business controlled by me and my son Jose despite a prohibition on outside business interests while in office. In fact, My husband is suffering from bronchitis and emphysema right now and he detained at the Veterans Memorial Medical Center in Quezon City in hospital prison outside Manila where the life of my husband is in danger.I will let you know that it is political motivated by Gloria Arroyo. Meanwhile,the government has said that it may drop rebellion charges against my husband allies Senator Juan Ponce Enrile and the former ambassador to the United State Ernesto Maceda because they were in the side of my husband,both men were later jailed by the government of Gloria Arroyo that they instigated a march on the presidential palace by 50,000 supporters of my husband.These are all political.All these are done to have my husband death. Presently life has not been easy for my children and I,my travelling documents have been seized by the government and they restricted me and my children to have access to my husband. Haven noticed the way things are going with us,my husband decided to let me know that he deposited some money with some Banks .These funds are presently deposited in his private Bank accounts, Three in whole, one in Europe,one in Central America and one in the Bahamas and all deposited in our name. The Fund in question is put all together exceed One hundred Million. 45 Million USD deposited with a Bank in CommonWealth of Dominica,79. Million USD with a Bank in Spain - and the Bahamas- 35. Million USD. As a prominent person here in Philippines I have no intention of getting involved in any criminality, and as such, no matter the gain I may stand to make I will not attempt to secure the Transfer of any of these sums if the process is not legal or is avoidable due to contravention of International Banking Statutes. I have in this regard studied the present status of the said funds against the order of the Government of Philippines as supported by the government of America, The Americas and Europe to freeze and terminate all Money Transfers to or From an account in our name. And I find it impossible to ensure the safe transfer of the Money in Bahamas, hence I will not attempt to.I have also found a way to tackle the Bank in Spain by using the service of a credit Commission who assist us in the lifting of the said fund from our bank PNB to the bank in Spain.Lastly,And I have reached an understanding with a Director of the bank in Commonwealth of Dominica and we have both devised a plan ( In which he is to play no active role). The above plans I mentioned are both legal, and feascible in so far as well as we can get the participation of a Foreign entity whether individual or corporate who would participate in the execution of our intention which I can safely declare to be risk free in all absolute. Your participation in this regards will entail that you enter into a trust agreement with me, wherein, in my capacity as Administrator of My Estate I will appoint you as Trustee and I, Trustor over the said sum, deposited at the bank in Dominica and the Bank in Spain. This agreement will form a legal basis for the Transfer of the amount to you, where as you will open an account with the bank of the same branch, because this is the only way we can execute a Transfer from the fund in the account without provoking an enquiry. You will noticed that when Monies are transfered from a Bank to another bank exceeding Hundred thousand American Dollars, the banks must report such transaction to the Apex Bank and if any of the parties whose itinerary is stated in the Bill of Transfer does not have clearance from the Government or is having such status as would ensure the money is siezed and the transfer will be aborted, Terminated and the fund confisticated, but if such transfer is done within the same bank ,the bank MAY NOT report such to the government Bank. Thus, if you were to execute an agreement of Trusteeship with me and open an account with the banks.I will have the banks wire the fund in our account into yours within the same bank and you will inturn wire the money to your oversea account in your country and hold the money in Trust under the terms of the agreement we will enter for a period that would not exceed four months. And then you will transfer the fund to a Bank in Malaysia where we are making arrangements to open a special account.
Now I want to start a new life with the money.I want to invest the money outside Philippines. I will want you to appreciate the fact that we are yet unaquainted, thus in your response a brief profile of yourself will certainly promote Trust on my side. I will stop here hoping to recieve an earnest positive response from you, where as I will implore you to handle this matter with utmost confidentiality and sincerity, so that we can have many other profitable interaction in the near future. I await your response. I would want us to be in partnership in any good business you may suggest in your country. Please handle this transaction with maturity and sincerity. Best Regards, Dr Mrs Luisa Pimental Estrada __________________________________________________________ Get your Private, Free Email from HTTP://www.DmailMan.Com
I'm looking for tips on sectioning 5 to 50 micron thick polymer films. Typically samples would be polyolefins, but sometimes (including my current set) they are nylons. I have a room temp microtome available, and have had *limited* success sectioning some nylons to around 2 microns, but sometimes need to go thinner.
I would appreciate any tips?embedding medium, sectioning conditions, etc.
Commercial responses are welcome off-list, both from vendors and service labs.
thanks,
Jim Passmore Sr. Analytical Chemist Cryovac Division Sealed Air Corporation james.passmore-at-sealedair.com
HI Dave, I've done immunfluor. of HA fixing with 2% pfa in the presence of 0.075% saponin. Never tried glut. Lee -- Leona Cohen-Gould, M.S. Sr. Staff Associate Director, Electron Microscopy Core Facility Manager, Optical Microscopy Core Facility Joan & Sanford I. Weill Medical College of Cornell University voice (212)746-6146 fax (212)746-8175
On Tuesday, May 20, 2003, at 09:51 AM, by way of Ask-A-Microscopist wrote:
} Question: My questions about an older Zeiss EM-9 TEM that I've been } working on for use in our High School science program. It's now fully } functional and working well. However, my most important concern is to } make sure that no X-rays are being emitted from around the gun. I had } assumed that a Geiger tube would detect the presence of X-rays } although I'm not sure that this is true. I've tried this method with } negative results, yet I do want to make 100% certain before I have } students learning to use this EM. } } Can you please advise?? } Dear Michael, A Geiger counter--the simple, hand-held type--should be sufficient. You could also use a hand-held ionization-chamber detector (one brand is called QT-pi), or you could put a film badge on the gun as a continuous monitor. Be sure to check all around to eliminate the possibility of directional x-ray leaks; e.g., between pieces of lead shielding. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
To keep the resin from splitting away from the film, you will need to section a small piece of the film that is surrounded by the resin. To do this, cut your polymer film into small triangles and then glue a thin wire with super glue to back end of the triangle. Put the pointed end into a beam capsule and wrap the end of the wire around the little piece of plastic that holds the cap on and try to wrap it tight enough to hold securely. You may have to bend the wire so that the pointed end is positioned properly for sectioning. Add some medium grade LRWhite and polymerize. We have cut both thick and thin sections from these blocks. Use formvar coated grids for thin sections.
Phoebe J. Doss Manager/Adjunct Instructor Electron Microscope Lab Department of Physiological Sciences 264 McElroy Hall Oklahoma State University Stillwater, OK 74078 Phone: 405-744-6765 Fax: 405-744-8263 http://www.cvm.okstate.edu/research/Facilities/EMLAB
In the footsteps of the ImageJ-mailinglist, I started a new mailinglist today for all Zeiss KS300/400 users among us. So if you use this program, and you would like to exchange your ideas/knowledge about the program and macro-editing in it, or you might have questions about the program or writing the macro's for image-analysis, I hereby invite you to join this mailinglist.
To subscribe, send a message to: KS300-subscribe-at-topica.com or click: http://www.topica.com/lists/KS300/subscribe/?location=listinfo
To send a message: KS300-at-topica.com
If you don't use the program, but you know some people who do, please forward this email to them. Just as the ImageJ-mailinglist, I think it might be a very helpful tool in improving the image analysis and knowledge and to get to know other persons who use these programs or to announce courses, workshops etc.
Also Zeiss LSM-macro-editors, or Axioplan-users are welcome to join the list.
I'm looking forward to your response. Best regards,
Sven Terclavers
LM/CLSM Microscopist Research Assistent Center for Transgene Technology and Gene Therapy Campus Gasthuisberg O&N Herestraat 49 3000 Leuven Belgium Tel: +32 (0)16 34 61 73 Fax: +32 (0)16 34 59 90 Email: Sven.Terclavers-at-med.kuleuven.ac.be Web: www.kuleuven.ac.be/mcm
The Lehigh Microscopy School. The Lehigh Short Courses on Electron Microscopy will be held this year (as they have for over 30 years) in June.
There are still vacancies in all of the courses. Please contact : Sharon Coe (610) 758 5133 Fax 610 758 4244 sharon.coe-at-lehigh.edu Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195
The Courses:
June 9 to 13 Scanning Electron Microscopy and X-ray Microanalysis A basic SEM course (With a one-day elementary introduction on June 8) June 16 to 20 A choice of 6 advanced courses on: Advanced SEM (Imaging) Advanced SEM (Analysis) Analytical TEM Characterization of Nanostructures Characterization of Particles AFM and other probe microscopies
Further details are at: http://www.lehigh.edu/~inmatsci/Microscourses.html or http://www.lehigh.edu/~inmatsci/shortcourses/Microscourses.html -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
I was hoping someone might be able to help me, as we have just lost our only spare domed glass bell jar (305mm x 356mm) for our Edwards coating unit, and we are being quoted $2747 to get a replacement. Does anyone have one they no longer need that we can buy from the at a reasonable cost.
Thanks
Steve Parry -- Steve Parry Centre for Microscopy & Microanalysis, (M010) The University of Western Australia, 35 Sterling Highway, Crawley, WA 6009, Australia.
MY DISCLAIMER!} If what follows does not stimulate a storm of protest from the physicists on the list, then it likely is almost accurate. I am a biologist, so there is always a danger that I have missed something that is important - especially when I make a suggestion for a 'cheap' survey as below.
In Pennsylvania, and in Colorado, I bet, one can get free copies of the state regulations, usually from the state "EPA", for radiation testing and responsibility. These regs will contain sections on radioactive materials as well as three types of radiation emitting equipment: medical X-Ray equipment, analytical X-ray equipment (such as x-ray diffractometers that use "high energy" x-ray generation equipment), and electron microscopes. Before you can determine your responsibility and that of your school, you have to get those relevant state regulations - even if they are difficult to understand.
In Colorado there is common sense in place. All you have to do is ascertain from the school district what IT will consider a proper "survey". However, it is clear that if the scope fails a formal (usually by a state-registered radiation physicist) survey that you will be in for some down time and likely a continuing survey program. On the other hand, you could preempt the problems by performing your own test first (as below) to determine that the scope is without a problem and THEN go thru the formal process to provide protection for all and limit cost to a one-time affair.
Seems to me you could do a satisfactory survey with Kodak 4489 film or strips of 35mm B&W film (that you had placed in a light-tight home made cardboard envelope with a piece of Al foil wrapped around half of the film. Place one or more of these light-tight packs over each of the column joints, at the top of the column where the cable joins, one over each of the viewing chamber windows and at the 'weak' points' of the camera system [which is situated in the most dangerous place for you [unless you are as old as I], AND most importantly around the high voltage tank on each face and at weak points on and around the top. I would run the scope for 24 hrs at the highest kV with the filament on. Oh yes. If there is a filament current monitor on the 9, you can expect more x-ray emissions the higher the gun dark current, especially if it exceeds 1uA (microampere). Also, if there had been or is a serious oil dispersion thru the system, that could contribute to higher emission currents and x-rays.
Most surveys that I do here are performed at 6" from the instrument. If you are clean up close, you should be clean elsewhere. Finally, the Al foil can overcome the problem of penetrance of high energy x-rays that may not register on the film by attenuating the power, or generating lower energy Al x-ray energies sufficient to permit formation of a latent image. In the absence of these 'harder' types of x-rays the Al protected film will serve, for each sheet/strip, as a negative control.
I have a small device named the "Radiation Alert INSPECTOR", manufactured by S.E. International, INC., PO box 39, 436 Farm Rd., Summertown, TN, 38483-0039. URL: http://www.seintl.com. This meter is marketed for the home owner who wants to monitor his/her basement for Radon, thus it is perfect for our purposes as well. [NO relationship beyond customer!]
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: michael_burnham-at-ceo.cudenver.edu [mailto:michael_burnham-at-ceo.cudenver.edu] Sent: Tuesday, May 20, 2003 12:52 PM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (michael_burnham-at-ceo.cudenver.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 20, 2003 at 09:14:34 ---------------------------------------------------------------------------
Email: michael_burnham-at-ceo.cudenver.edu Name: Michael Burnham
Organization: Kent Denver School
Education: 9-12th Grade High School
Location: Denver, Colorado
Question: My questions about an older Zeiss EM-9 TEM that I've been working on for use in our High School science program. It's now fully functional and working well. However, my most important concern is to make sure that no X-rays are being emitted from around the gun. I had assumed that a Geiger tube would detect the presence of X-rays although I'm not sure that this is true. I've tried this method with negative results, yet I do want to make 100% certain before I have students learning to use this EM.
Can you please advise??
Thank you greatly,
Michael Burnham Kent Denver School Chair Science Dept. 4000 E. Quincy Ave Englewood, CO 80110
Hello Else, As you have probably noticed the inside walls of the ion mill get coated by a film deposit over periods of use. These deposits can flake off and find there way in the whisper lock mechanism. The piston rod or more likely the seals in the whisper lock mechanism might be dirty. We periodically dismantled the whisper lock mechanism, cleaned it and replaced the seals. This kept the rotation smooth and corrected vacuum leaks that would develop. You might try to see if that would fix the problem. You could also inspect the motor/gears for possible damage while disassembled. Tyrone Daulton
"Else Breval (by way of MicroscopyListserver)" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } I am a beginner, I recently subscribed. } } I have a problem, but I do not know whom to ask. } } I am ion millimg on a GATAN Dual Ion Mill, Model 600A. } The machine is very old, but good, when it works. } } I have the problem that the stage, which usually rotates as soon as it } is brought in the lower position, does not rotate as it should. } } Can anyone give me a tip why? and what can I do about it? } } Else Breval } Penn State University } Materials Research Institute
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Facility Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
Ambient temperature microtomy of polyolefins is a difficult task and one that gives generally poor results at best. Although apparently acceptable sections may be obtained for optical microscopy, one should still be very conservative in the interpretation of morphology.
I recommend that you spring for a cryogenic ultramicrotome if you plan to continue such work. This is the only way to get good quality sections from polyolefins. When sectioning polymers, the temperatures of the sample and knife need to be below the glass transition temperature of all components in the film to avoid deformation during cutting. Polypropylene should be cut below -10C; polyethylenes of any type below -120C.
Feel free to contact me off-line to discuss this further.
Cheers,
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
"James.Passmore-at-sea ledair.com" To: microscopy-at-sparc5.microscopy.com cc: Subject: polymer microtomy 05/21/03 08:58 AM
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Hello everyone.
I'm looking for tips on sectioning 5 to 50 micron thick polymer films. Typically samples would be polyolefins, but sometimes (including my current set) they are nylons. I have a room temp microtome available, and have had *limited* success sectioning some nylons to around 2 microns, but sometimes need to go thinner.
I would appreciate any tips?embedding medium, sectioning conditions, etc.
Commercial responses are welcome off-list, both from vendors and service labs.
thanks,
Jim Passmore Sr. Analytical Chemist Cryovac Division Sealed Air Corporation james.passmore-at-sealedair.com
Our Zeiss 10C TEM has a problem to take pictures. After I started to take picture, even the screen was down in a normal position no beam light was seen for a while (say 3-7 seconds). The "exposured film" was nothing but some shadows with different darkness after development. The shadows look like you were trying to figure out the exposure time when you are printing a micrographs in the dark room. At first we thought it might be shuttle jammed, then we took it out of column, it worked fine outside of the column. However, the same problem occurred, namely no beam light was seen for a while. The shuttle problem might be excluded. We have no idea what problem could be. Any suggestions??
I appreciate your help in advance.
Haixin Xu Department of Biological Sciences University of Maryland Baltimore County Maryland USA
I've had good success using a room temperature microtome, by freezing the specimen before cutting sections. The results are not as consistent as one would expect with a cryomicrotome, but might provide suitable results, especially if a cryomicrotome is not available. I freeze the sample while it is mounted in the chuck by holding a cotton swab saturated with liquid nitrogen close to, but not touching, its surface.
James Martin Orion Analytical, LLC A Materials Analysis and Consulting Firm www.orionanalytical.com 413-458-0233
----- Original Message ----- } From: {"James.Passmore-at-sealedair.com"-at-sparc5.microscopy.com} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, May 21, 2003 9:58 AM
} Pacific Northwest Microscopy Society announces agenda for its upcoming annual meeting at PNNL in Richland, WA on May 29 and 30. } } Thursday May 29, 2003 } (Registration and badging starting at 11 am in EMSL lobby) } 1:00 pm Welcome note - Alice Dohnalkova, Jim Young } 1. Focused Ion Beam (EMSL Auditorium) } 1:05 FIB - Specimen preparation for Everyting - Lucille Giannuzzi, University of Central Florida } and FEI company - MSA invited speaker } 1:45 In situ sample preparation and HR SEM-STEM analysis - Matthew Weschler, FEI Company } 2. Advances in Analytical Techniques in Material Science (EMSL Auditorium) } 2:10 Backscatter Electron Imaging of Microstructural Constituants in Steels - } James H. Steele } 2:30 Nanometer Resolution Analyses of Crack Tips in a field-emission-Gun TEM } - Larry Thomas, PNNL } 2:50 The right tool for the right job - the latest generation of x-ray detectors for } EDS - Del Redfern, EDAX } -------- Break -------- } 3:20 Use of Electron Energy-Loss Near Edge Fine Structure in the Study of Phases } from the Sr/TRU Precipitation Process - Edgar Buck, PNNL } 3:40 Projection X-ray microscopy in the SEM - Les A. Brownlow, XRT Limited } 4:00 Helical Nanowires and Nanosprings - Hai-Feng Zhang, WSU } 3. Flow Cytometry - (Conference room 1075) } (simultaneous with Material science platform) } 2:10 Basic Flow Cytometry as a Comparative Method for Imaging and } Fluorescence - Randall W. Smith, Portland State University } 2:40 Construction of the first human nonimmune scFv library for yeast display } and the utility of flow cytometric-based selections - Robert Siegel, Molecular } Biosciences, PNNL } 3:00 - 4:00 pm Demo on a practical flow cytometry application - Jeff McLean, } Environmental Microbiology, PNNL } Participants will gather in laboratory 115 in building 331; anticipated return to } EMSL at 4pm. } Poster Session (Conference room 1077) } 4:30-5:30 pm Fifteen posters on topics Advances in microscopy technologies, Fluorescence and Laser probe microscopy, Analytical applications in material science, Environmental applications, and Microscopy education will be presented. Judges will select three student posters to be awarded cash prizes. } Expo (Conference room 1077) } The exhibition of literature and selected instruments presented by companies manufacturing or marketing microscopy-related products will be held during the meeting in room 1077. Complimentary refreshment will be served. } Evening Social } 6 pm - ? } PNMS members and registered meeting participants are welcome to attend an evening reception at Gordon Brother} '} s winery in Pasco and enjoy catered dinner, local wine tasting and a talk on Geological history of the Columbia river given by Steve Reidel. The winners of the poster session will be announced and awards given to the three best presentations. Busses are leaving EMSL at 6pm and returning back to EMSL after the social. Additional tickets can be purchased at registration. } } Friday May 30, 2003 } 4. Laser Microscopy (EMSL Auditorium) } 9:00 Getting as much information as possible using nonlinear optical microscopy - } Gary Holtom, PNNL } 9:35 Imaging signal transduction in live cells of the neuroendocrine system - Anda } Cornea, OHSU } 9:55 Three Dimensional Finite Element Method Simulations of the Aperture-less, } AFM-Tip Enhanced Near-Field SERS Microscopy - M. Micic, PNNL } ------ Break ------ } 5. 3-dimensional Imaging and Reconstruction } 10:20 3-d Imaging and Cell Reconstruction by EM Tomography: a primer - Alice } Dohnalkova, PNNL } 10:40 Image Processing, Segmentation, and Feature Extraction Algorithms} } Applied to Electron Tomography Data - Harold Trease, PNNL } 11:00 High speed 3-D Visualization of Signal Transduction - Steve Wiley, PNNL } ------ Break ------ } 6. Environmental Applications (EMSL Auditorium) } 1:00 Cathodoluminescence Imaging and Spectroscopy in the SEM - Applications } from Mineralogy, Semiconductors and Biomedical Samples - Paul Mainwaring, } Gatan Inc. } 1:20 Examination of Gas Hydrates in Sediments with Environmental Scanning } Electron Microscopy - Jim Young, PNNL } 1:40 Identification of Indoor Particles by Analytical Light Microscopy - E. Russ } Crutcher, Microlab Northwest } 2:00 Probing Heterogeneous Chemistry of Individual Atmospheric Particles Using } Electron Microscopy Techniques - Alexander Laskin, PNNL } } }
Some laboratory glass blowers can make new bell jars to your specs. This should save you some money. Alternatively, check out the used equipment suppliers.
Good luck.
"The statements and opinions expressed here by Gary M. Brown represent neither those of ExxonMobil Corporation nor its affiliates."
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
Steve Parry {sparry-at-cmm.uwa.ed To: Microscopy-at-sparc5.microscopy.com u.au} (by way of cc: MicroscopyListserv Subject: Coating Unit Bell Jar er)
05/22/03 07:02 AM
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Can you help??
I was hoping someone might be able to help me, as we have just lost our only spare domed glass bell jar (305mm x 356mm) for our Edwards coating unit, and we are being quoted $2747 to get a replacement. Does anyone have one they no longer need that we can buy from the at a reasonable cost.
Thanks
Steve Parry -- Steve Parry Centre for Microscopy & Microanalysis, (M010) The University of Western Australia, 35 Sterling Highway, Crawley, WA 6009, Australia.
The schedule of abstracts for The Microscopical Society of Canada meeting in Vancouver is now on the website: http://www.emlab.ubc.ca click on MSC logo. Elaine
-- Dr. Elaine Humphrey Director, BioImaging Facility First Vice President, Microscopy Society of Canada University of British Columbia 6270 University Blvd, mail-stop Botany Vancouver, BC CANADA, V6T 1Z4 Phone: 604-822-3354 FAX: 604-822-6089 e-mail: ech-at-interchange.ubc.ca website: www.emlab.ubc.ca
Try Dunniway Stockroom, www.duniway.com for "surplus" vacuum equipment of all kinds. Phone 1-650-969-8811, email info-at-dunniway.com in Mountain View California. I have purchased at very reasonable cost both used and new vacuum equipment from them and I am a satisfied customer. Usually they have some bell jars available.
Ron Vane XEI Scientific
Disclaimer: No financial interest, just a customer.
----- Original Message ----- } From: "Steve Parry (by way of MicroscopyListserver)" {sparry-at-cmm.uwa.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, May 22, 2003 5:02 AM
How did you get on with the mouldy old bread over a wet Bank Holiday? "Mouldy Old Dough" was a hit the early seventies for Lieutenant Pigeon, I believe.
Dave
On Mon, 5 May 2003 14:26:30 +0100 (BST) "Robert H. Olley" {r.h.olley-at-reading.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } This is to thank everybody who replied with a wealth of ideas concerning } Roots and SEM. If haven't had any feedback yet from our soil science } department, so I can't as yet report any progress on that front. } } However, in regard to moulds, I inadvertently (honest!) let some bread go } mouldy recently. So I've looked through all the suggestions. For fixing, } one can't slowly go up through the progressions of increasingly } concentrated ethanol, since the bread will go intractably soggy (does } anyone remember the song "Mouldy Old Dough"?) Neither do I want to hit } the mould with neat alcohol. So what I'm doing, as a first stage at } least, is to put thin mouldy crusts of the bread over methanol in a vapour } tank, and letting it stand over the long weekend (we British are having } our May Day Holiday on Monday 5th!). If anything useful results, I'll } keep you posted. } } Bye for now, } } +-----------------------------------------+ } Robert H.Olley } J.J.Thomson Physical Laboratory } University of Reading } Whiteknights } Reading RG6 6AF } England } +----------------------------------------+ } Phone:+44 (0) 118 9318572 } Fax: +44 (0) 118 9750203 } University internal extension 7867 } Email: R.H.Olley-at-reading.ac.uk } URL: http://www.reading.ac.uk/~spsolley } +----------------------------------------+ } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
A position for an electron microscopy (EM) research assistant in the Shaham lab at Rockefeller University is available immediately. The successful applicant will actively participate in ongoing and new projects to understand the cellular basis of apoptosis and glia-neuron communication in C. elegans. Previous EM experience, including serial reconstructions and 3-D computer imaging is a plus, however, candidates with demonstrated interest in EM will be considered as well.
We offer an excellent benefits package, competitive salary, & tuition reimbursement. For immediate consideration, email or fax resume to The Rockefeller University at FAX (212) 327-7079 or EMAIL recruit-at-rockefeller.edu.
An Equal Opportunity/Affirmative Action Employer. The Rockefeller University appreciates all responses and will contact candidates selected for further consideration.
We have had several requests concerning the use of Mercox in plants. As far as we know our customers presently use Mercox on animals. If there is anyone out there with experience in corrosion casting with plants we would appreciate your input.
Thank you,
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
I have seen two types of Edwards 306A units with different bell jars. Do you mean you have a 306 type? Type 1 has a bell jar that is a cylinder with a gasket at both ends. At the top is a 'flip top' metal lid that opens into the interior of the glass cylinder bell jar. Type 2 is a regular dome shaped bell jar and could be removed manually. Type 3 had an implosion shield and hoist to lift up the dome shaped glass bell jar. Edwards made other type of evaporators.
Ladd or Lesker may be able to order you one. Try Lesker.com for about $800 for a 12 inch by 18 inch glass domed bell jar. You can always have a glass blower cut off a few inches to make 14 inches and then grind a sealing surface for you. http://www.lesker.com
I tried many L gaskets a few years back and Ladd had the nicest one I could find without cracks or defects in the sealing surface. They sell various Pyrex® glass bell jars too. http://www.laddresearch.com/Key_Products/VacEvap/VacEvap/Bell_Jars___Support /bell_jars___support.html
Hope this helps. I don't work at these places or have any interest in them.
Paul Beauregard Senior Research Assocaite PPG Industries Monroeville Technical Center Monroeville, PA 15146 724-325-5131
} Can you help?? } } I was hoping someone might be able to help me, as we have just lost } our only spare domed glass bell jar (305mm x 356mm) for our Edwards } coating unit, and we are being quoted $2747 to get a replacement. } Does anyone have one they no longer need that we can buy from the at } a reasonable cost. } } Thanks } } Steve Parry } -- } Steve Parry } Centre for Microscopy & Microanalysis, (M010) } The University of Western Australia, } 35 Sterling Highway, Crawley, } WA 6009, Australia. } } Fax: +61-8-9380-1087 } Email: sparry-at-cmm.uwa.edu.au } Phone: +61-8-9380-8057 [24 hour voicemail attached] } } }
Dear Listers, I've been trying to search microscopy websites for those plastic screw-top vials to store unused objectives in, but I can't seem to find them anywhere. Are they just not used anymore, or am I looking in the wrong place?
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This is a TEM job at the Burnham Institute in La Jolla, CA Please reply to Dr. Dorit Hanein (e-mail at the bottom of the announcement).
Position Open: Electron Microscopist Research Assistance, to start June, 2003. (http://www.burnham-inst.org).
Requirements include working knowledge of high resolution TEM and a BA/BS or equivalent experience in biological or material science, or bioengineering. Responsibilities include negative stain sample preparation and electron microscopy, vitreous ice sample preparation and electron cryo-microscopy, some microscope maintenance and standard biochemical analyses.
Application should include a statement of career goals and addresses for three references. Salary is competitive and commensurate with experience. EOE.
Send applications to: Dr. Dorit Hanein, e-mail dorit-at-bunham.org
The Analytical Imaging Facility of the Albert Einstein College of Medicine, a core microscopy facility, is expanding its technical staff. We have two openings for experienced biological microscopy technicians.
Position 1: Digital Flourescence and Confocal Microscopy Technician
Position 2: Electron Microscopy Technician
Details for these positions can be found in the May/June issue of Microscopy Today, page 41. Please note: The email address for responding is incorrect. For consideration send your CV to ddamiani-at-aecom.yu.edu
The positions are also posted on the MSA Placement Office listing:
http://www.msa.microscopy.com/PlacementOffice/JobListings.html **************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
Does anyone have another source for the 12mm diameter carbon conductive tabs? I used to get 16084-1 from Pella but now they are standard double thickness and not useable. They are not as sticky and do not bend.
Hi Pauline, Here's what I would try to find, though I would always remember to include sufficient size that I can also include a small pack/pillow of silica gel desiccant - I use NO substitutes for this material. BTW, to dehydrate this material (silica gel!), I place it in my EM film plate degasser for a few days - thus, any vacuum device pumped to 30in of Hg will do. No need to keep the pump on or to include P2O5, just the 30in vac is sufficient for a considerable amount. For those with confidence, for some objectives I have recycled screw-cap plastic tritium shipping containers - also other decontaminated and de-labeled radioactive containers. Mine are all blue or orange and work beautifully for my old oculars and objectives - when I, like you, can't find a REAL objective or ocular container.
Try: http://www.tri-esssciences.com/plasticjars.htm [NO knowledge of this company or relationship to it OR manufacturer, etc.]
I do not ever enclose objectives without some silica gel as a mild desiccant.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: pauline yu [mailto:pcy-at-usc.edu] Sent: Friday, May 23, 2003 5:59 PM To: Microscopy
Dear Listers, I've been trying to search microscopy websites for those plastic screw-top vials to store unused objectives in, but I can't seem to find them anywhere. Are they just not used anymore, or am I looking in the wrong place?
Please let us know what size 'objectives' you are trying to store and we'll see if we can assist.
John Arnott
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "pauline yu" {pcy-at-usc.edu} To: "Microscopy" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 23, 2003 5:59 PM
Steve,
As Paul mentioned we have a 12" x 18" domed bell jar. It would be quite expensive to special order a 12" x 14" . If you do cut it down the L-gaskets would probably help maintain the vacuum.
John
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Beauregard" {beaurega-at-westol.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 23, 2003 5:50 PM
Hi Steve,
I have seen two types of Edwards 306A units with different bell jars. Do you mean you have a 306 type? Type 1 has a bell jar that is a cylinder with a gasket at both ends. At the top is a 'flip top' metal lid that opens into the interior of the glass cylinder bell jar. Type 2 is a regular dome shaped bell jar and could be removed manually. Type 3 had an implosion shield and hoist to lift up the dome shaped glass bell jar. Edwards made other type of evaporators.
Ladd or Lesker may be able to order you one. Try Lesker.com for about $800 for a 12 inch by 18 inch glass domed bell jar. You can always have a glass blower cut off a few inches to make 14 inches and then grind a sealing surface for you. http://www.lesker.com
I tried many L gaskets a few years back and Ladd had the nicest one I could find without cracks or defects in the sealing surface. They sell various Pyrex® glass bell jars too. http://www.laddresearch.com/Key_Products/VacEvap/VacEvap/Bell_Jars___Support /bell_jars___support.html
Hope this helps. I don't work at these places or have any interest in them.
Paul Beauregard Senior Research Assocaite PPG Industries Monroeville Technical Center Monroeville, PA 15146 724-325-5131
} Can you help?? } } I was hoping someone might be able to help me, as we have just lost } our only spare domed glass bell jar (305mm x 356mm) for our Edwards } coating unit, and we are being quoted $2747 to get a replacement. } Does anyone have one they no longer need that we can buy from the at } a reasonable cost. } } Thanks } } Steve Parry } -- } Steve Parry } Centre for Microscopy & Microanalysis, (M010) } The University of Western Australia, } 35 Sterling Highway, Crawley, } WA 6009, Australia. } } Fax: +61-8-9380-1087 } Email: sparry-at-cmm.uwa.edu.au } Phone: +61-8-9380-8057 [24 hour voicemail attached] } } }
If you want to image the particles directly you need to do transmission electron microscopy. A FEG-SEM might work but a TEM can also do some electron diffraction structural determinations on the ultimate or primary particles. If you have a mixture of phases in the nanoparticles, you should try X-Ray Diffraction on the powder to show the different phases, if present.
If you are dealing with agglomerated or aggregated finer primary particles, this will be very obvious in a TEM.
Direct observations on a TEM grid with oil present can be done. Also one can extract the oil after the dispersion is on the grid. One skilled in the art of TEM and particle dispersions will be able to extract the oil from the grid dispersion without producing flocculation effects.
Don't ignore doing nitrogen BET determinations on the powders. To extract the particles from the oil for BET, heat it up gradually to about 525°C in nitrogen and then switch to air at 525°C. A TGA savvy person can help here and can actually isolate enough material for a TEM dispersion but BET will require about 50-300 mgs net. For the BET required amounts, use a heated tube furnace and combustion boat. I am assuming the nanoparticles were made by plasma and have no molecular roughness or microporosity in them that will raise the BET surface area value. Anyway, 50 nm smooth ultimate particles with a density of, say, 3.65 will give a BET of about 33 m²/gram. 11 nms ultimates will yield a BET of about 150 m²/g. 300 nm ultimates (your top range) will give a BET of about 6 m²/g. I can email you the exact assumptions and a simplified formula separately, if you are interested.
You can determine the primary or ultimate particle size distribution of small aggregates by interactively drawing diameters across the ultimate particles on the dispersed and finest aggregates you see. Use an on-line TEM and an image analysis program for this mouse intensive work.
The presence of oil means using microtomy is probably eliminated for evaluating aggregate sizes. It's possible but doubtful. One microtome pitfall would be that you can cut the real aggregate structures in half with the knife edge. One law of microtoming says something like this. As the thickness of the thin section approaches the ultimate particle size diameter, aggregates and ultimate particles that are agglomerated tend to artificially look more dispersed and smaller.
Hope this helps you out and gives you a few more options.
Paul Beauregard Senior Research Associate Greensburg, PA USA
} } Email: daniel.smith-at-liquidminerals.com } Name: Daniel Smith } } Organization: Liquid Minerals Group, Inc. } } Education: Graduate College } } Location: Houston, TX USA } } Question: I am interested in measuring particle size of metal oxide } particles suspended in a process oil. The particles are on the order } of 1 - 300 nanometers. I have used indirect methods (DLS, etc.), but } I would really like to take photos of the particles and determine } particle size, crystal structure, agglomeration and other properties. } } Any help would be appreciated. } } --------------------------------------------------------------------------- } } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rene.oberli-at-strasbourg.gm.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May 27, 2003 at 09:15:51 ---------------------------------------------------------------------------
Organization: process enginer at general motors strasbourg france
Education: 9-12th Grade High School
Location: strasbourg France
Question: I would like to make digital pictures with Canon G2 photo camera thru a WILD M3 (6 to 40x power, no zoom 1X front lens)with a LEiCA trinocular head up to now I was not able to get a full frame picture I also tried with special lens ( 0,6x )from LEICA or NIKON between the output tube and the G2 without success; the sharp image is circular and not full frame.This assembly works fine with a NIKON coolpix 995.Why this dont work with CANON G2 with or without zooming the G2? Canon-Europe is no help from a contact with them they dont know or dont want to help!
I am trying to figure a way to determine whether there are any birefringent particles ranging in size from 100nm to 3 micrometers in a whole blood sample, maybe 1mL volume. I don't have to preserve the blood components, maintain the structural integrety of the RBCs, WBCs, platelets, etc. I can not use any solvents, only aqueous solutions and I can't have the pH go below 3. What makes this more difficult is that the particles may be few in number, very few!!!
Would there be some way to break up the cells, digest the membranes, and wash away the blood material so that I could filter out the particles? If I did, I would have to separate the buffy coat and the plasma first, then go after the RBCs. I could check the plasma for particles separately and I know that the macrophages etc would have already taken up some of the particulate but that doesn't matter. Would SEM be the best approach? I would think so to see the very small particles. The particles are also reportedly fluourescent but excited by UV (which I don't have on my confocal). Would using a gradient with centrifugation be of use to separate out the particles? But then the particles may stick to the cells. Do you know of anyone who has done this type of thing and point me to any publications. I will be doing a literature search starting tomorrow as this just came up today.
Thanks for any ideas you might be able to pass along!!!
The ones I got from SPI works fine. Here is their contact details. Sadly I do not have personal interest in this company and will continue being overworked and underpaid.
Structure Probe, Inc. / SPI Supplies ·Street address for UPS, FedEx, DHL, other couriers, trucks 569 East Gay Street West Chester, PA 19380 ·Mailing address P.O. Box 656 West Chester, PA 19381-0656 USA
Phone: 1-(800)-2424-SPI Toll-free from USA/Canada 1-(610)-436-5400 FAX: 1-(610)-436-5755 E-mail: spi3spi-at-2spi.com
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, May 27, 2003 6:22 PM To: MSA listserver
Does anyone have another source for the 12mm diameter carbon conductive tabs? I used to get 16084-1 from Pella but now they are standard double thickness and not useable. They are not as sticky and do not bend.
This is a good question and hopefully someone can answer it. We buy our tabs from ProSciTech, trade name "Leit-tabs" I believe. About 12 months ago the tabs changed, they were thicker, stiffer and less sticky, as Gary notes, and in addition nowhere near as conductive. When you rub off the "sticky" there is a stiff polymer film in the middle. Alternative supplies of sensibly conducting tabs would be welcome.
} Does anyone have another source for the } 12mm diameter carbon conductive tabs? } I used to get 16084-1 from Pella but } now they are standard double thickness } and not useable. They are not as sticky } and do not bend.
You could ask Taab or Agar Scientific Ltd. (both UK) about their current stock. I prefer the thin bendy tabs but my most recent purchase contained the thick ones. I read on this list that some thick ones contain cardboard!
Dave
On Tue, 27 May 2003 09:22:26 -0700 Gary Gaugler {gary-at-gaugler.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Does anyone have another source for the } 12mm diameter carbon conductive tabs? } I used to get 16084-1 from Pella but } now they are standard double thickness } and not useable. They are not as sticky } and do not bend. } } Ideas?? } } gary g. } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
The tabs from Electron Microscopy Sciences (www.emsdiasum.com) are double thickness also so they do not bend but are still very sticky. (Cat# 77825-12) You can also try SPI supplies (www.2spi.com) Good luck! -Amanda
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Tuesday, May 27, 2003 12:22 PM To: MSA listserver
Does anyone have another source for the 12mm diameter carbon conductive tabs? I used to get 16084-1 from Pella but now they are standard double thickness and not useable. They are not as sticky and do not bend.
Ya, my last batch from 2 different suppliers is as you describe below. Unfortunately I purchased a whole years supply. Incidentally, has anyone had problems with the surface cracking during/after coating. This is a recent development, and I pretty much didn't care, but a user tacked down a delicate sample and it cracked with the tab. I don't have this problem with the dwindling supply of old tabs or with the tape roll.
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } Bryony James {b.james-at-auckland.ac.nz} 05/28/03 02:36AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
This is a good question and hopefully someone can answer it. We buy our tabs from ProSciTech, trade name "Leit-tabs" I believe. About 12 months ago the tabs changed, they were thicker, stiffer and less sticky, as Gary notes, and in addition nowhere near as conductive. When you rub off the "sticky" there is a stiff polymer film in the middle. Alternative supplies of sensibly conducting tabs would be welcome.
} Does anyone have another source for the } 12mm diameter carbon conductive tabs? } I used to get 16084-1 from Pella but } now they are standard double thickness } and not useable. They are not as sticky } and do not bend.
Hmmm. A hypotonic solution (hypotonic as compared to plasms, water or very dilute buffer) will cause the red cells (and the WBCs too I suppose) to swell and burst. Knowing the composition of the 'particles' you are looking for might help.
Geoff
Damian Neuberger wrote:
} Hi all, } } I am trying to figure a way to determine whether there are any birefringent } particles ranging in size from 100nm to 3 micrometers in a whole blood } sample, maybe 1mL volume. I don't have to preserve the blood components, } maintain the structural integrety of the RBCs, WBCs, platelets, etc. I can } not use any solvents, only aqueous solutions and I can't have the pH go } below 3. What makes this more difficult is that the particles may be few in } number, very few!!! } } Would there be some way to break up the cells, digest the membranes, and } wash away the blood material so that I could filter out the particles? If I } did, I would have to separate the buffy coat and the plasma first, then go } after the RBCs. I could check the plasma for particles separately and I } know that the macrophages etc would have already taken up some of the } particulate but that doesn't matter. Would SEM be the best approach? I } would think so to see the very small particles. The particles are also } reportedly fluourescent but excited by UV (which I don't have on my } confocal). Would using a gradient with centrifugation be of use to separate } out the particles? But then the particles may stick to the cells. Do you } know of anyone who has done this type of thing and point me to any } publications. I will be doing a literature search starting tomorrow as this } just came up today. } } Thanks for any ideas you might be able to pass along!!! } } Damian Neuberger } } } } } } }
-- -- ********************************************** 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 **********************************************
I agree that these double thick tabs that have quietly replaced the thin and flexible tabs we all came to love are the pits. Glad to hear there are others that don't like them either. As an example of why they won;t work for me, I cut C tabs into thin strips and use as "ground straps" on SEM samples. Double thick are inflexible and won't work for that. I'd really love to hear from a vendor as to why we need the new thick style!
So, here's the good news. E. Fullam sells 12mm carbon tabs that are thin, flexible and reasonably sticky. The part no is 14833, I called this morning and they have 27 in stock.
SPI's #5077 is a close second, but still too thick for me.
Pella's carbon tabs are like cardboard. Unusable.
Note, this is all personal preference, I ran no tests. I am a customer of the companies I mentioned.
Owen Mills Owen P. Mills Electron Optics Engineer 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
At 09:22 AM 5/27/2003 -0700, Gary Gaugler wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer 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
At ~2000g, all the cells will sediment. What about the particles? If the particles are less dense than hemoglobin, then layer a slightly less dense than particles but more dense than hemoglobin, hemolyze blood with distilled water, and spin. Your particles should 'fall' into 'dense' layer at bottom. On the other hand, your particles might be more buoyant than the cells, in which case you win that way. In the absence of answers to my questions, you are left with a manageable experiment anyway, especially if the particles start out in a jar on the shelf. If they are manufactured in the body, i.e., crystals, then you must do it the hard way. An Airfuge (Beckman) will sediment hemoglobin, but not certain mycoplasma-like organisms, and it will work with really small volumes. Even if the particles sediment faster than the hemoglobin, the hemoglobin isn't birefringent. With 1ml to start, you have a lot of flexibility.
Once you can use sedimentation, you can concentrate the stuff. In the end, by my estimate, the best bulk dehydrant of stuff in a dialysis bag is PEG (polyethylene glycol).
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Damian Neuberger [mailto:neuberger1234-at-attbi.com] Sent: Tuesday, May 27, 2003 10:40 PM To: Microscopy-at-sparc5.microscopy.com
Hi all,
I am trying to figure a way to determine whether there are any birefringent particles ranging in size from 100nm to 3 micrometers in a whole blood sample, maybe 1mL volume. I don't have to preserve the blood components, maintain the structural integrety of the RBCs, WBCs, platelets, etc. I can not use any solvents, only aqueous solutions and I can't have the pH go below 3. What makes this more difficult is that the particles may be few in number, very few!!!
Would there be some way to break up the cells, digest the membranes, and wash away the blood material so that I could filter out the particles? If I did, I would have to separate the buffy coat and the plasma first, then go after the RBCs. I could check the plasma for particles separately and I know that the macrophages etc would have already taken up some of the particulate but that doesn't matter. Would SEM be the best approach? I would think so to see the very small particles. The particles are also reportedly fluourescent but excited by UV (which I don't have on my confocal). Would using a gradient with centrifugation be of use to separate out the particles? But then the particles may stick to the cells. Do you know of anyone who has done this type of thing and point me to any publications. I will be doing a literature search starting tomorrow as this just came up today.
Thanks for any ideas you might be able to pass along!!!
If you are examining liver for alpha-1-trypsin, then I would recommend the following text. Although it is a bit dated it has beautiful full page micrographs.
Phillips, MJ, S. Poucell, J. Patterson, P. Valencia.1987. The Liver: an ultrastructural atlas and text of liver diseases. Raven Press, NY, NY, 585 p.
} } } Garry Burgess {GBurgess-at-exchange.hsc.mb.ca} 05/21 11:49 AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Does anyone have any references which show micrographs of alpha-1-trypsin as seen in the electron microscope?
Of late there has been discussion on the carbon tabs and the change in the consistency and thickness of the tabs. This is true that approximately 6-8 months ago the raw material that had been used over many years to make these tabs become obsolete. The manufacturers sourced these materials from the same manufacturers of type writer ribbon however when we all stopped using ribbon we lost the source for the carbon raw. The new material is a bit thicker than and not as sticky as the original but for most it is fine and works adequately. Alternatively we here at Electron Microscopy Sciences has found a new source from the Far East that has raw material very similar to the original which produces very thin tabs with the same sticky power as the old original. The problem is the new raw material is double the cost of what was. We do have ample stock of it for those of you whom are looking for the thin material. The good news is doing a search out there on all of the thin material our price of $60 per 200 tabs seems to be very low. Please do not hesitate to contact me if you have any questions I look forward to hearing from you. Sincerely,
Stacie Kirsch Electron Microscopy Sciences 215-646-1566
I am looking for some Mylar (polyethylene I think) sheets, clear not 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any ideas who/where sells this stuff?
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
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From root Wed May 28 20:46:56 2003 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id UAA27563 for dist-Microscopy; Wed, 28 May 2003 20:18:36 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id UAA27560 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Wed, 28 May 2003 20:18:05 -0500 (CDT) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id UAA27553 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 28 May 2003 20:17:54 -0500 (CDT) Mime-Version: 1.0 X-Sender: zaluzec-at-ultra5.microscopy.com (Unverified) Message-Id: {p05111a00bafb0eeeabff-at-[206.69.208.21]}
Hello.
I am doing some primary research of AFMs. I can cacross two scanning modes which I am unfamilaiar with, and which I cannot find a good source of information on: curren timaging and phase imaging. I have a good idea of what they do, but i wonderign if you could inform me of good descriptiuons of eahc mode or coudl tell me where I could find such information. Thank you very much for your time,
Joseph D. Matchett
From root Wed May 28 21:15:09 2003 Return-Path: {Microscopy-request-at-sparc5.microscopy.com} Received: (from daemon-at-localhost) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id UAA27596 for dist-Microscopy; Wed, 28 May 2003 20:19:58 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id UAA27588 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Wed, 28 May 2003 20:19:27 -0500 (CDT) Received: from [206.69.208.21] (mac21.zaluzec.com [206.69.208.21]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id UAA27578 for {Microscopy-at-MSA.Microscopy.Com} ; Wed, 28 May 2003 20:19:15 -0500 (CDT) Mime-Version: 1.0 X-Sender: (Unverified) Message-Id: {p05111a02bafb0f38bd7e-at-[206.69.208.21]}
I would greatly appreciate your help in dealing with the following problem. We have a set of samples from an experiment on rats performed yesterday (Tuesday, May 27). The lung and aorta samples were divided into two parts each: one, for embedding in EMbed 12, was brought to the step of washing after osmium tetroxide staining and the other, for cryosectioning, was brought to the step of infiltration with PVP and sucrose. The first set of samples is currently in 0.1 M sodium cacodylate buffer, pH 7.4, and the other in the PVP with sucrose in phosphate buffer pH 7.0. However, there is a delay in delivery of the embedding kit and aluminum specimen pins. Most likely, both will arrive here on Monday, June 2nd. Is it safe to keep the samples at this stage of preparation until Monday? If yes, at what temperature?
I don't see big problem. Keep it refrigerated +4oC. Extended wash in cacodylate may wash out some components of the cell. You may expect some artefacts from that. If your samples for cryo-EM were fixed with formaldehyde, you know: formaldehyde fixation is reversible. So, extended wash after formaldehyde may cause "less" fixation. Sergey
At 06:13 PM 5/28/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
FWIW, Mylar and polyethylene are quite different. Anyway, If you can use it, I have some I believe to be mylar, but it is only 1/2 mil thick (0.0005").
Woody
Woody White BWXT Services: http://www.bwxt.com/bwxt.html My Site: http://woody.white.home.att.net
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, May 28, 2003 12:58 PM To: Microscopy
Dear List:
I am looking for some Mylar (polyethylene I think) sheets, clear not 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any ideas who/where sells this stuff?
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
How about a couple of part numbers for the old and new tabs: Thick 12 mm: 77825-12 ? (Our order on this number was filled with thicker tabs.) Thin 12 mm: ?????
Thick 25 mm: 77825-25 ? $$ Thin 25 mm: ????? $$
Thanks,
Paul
-----Original Message----- } From: Stacie Kirsch [mailto:Stacie-at-ems-secure.com] Sent: Wednesday, May 28, 2003 11:31 AM To: Microscopy-at-sparc5.microscopy.com Cc: spb-at-mwrn.com
Dear Microscopists,
Of late there has been discussion on the carbon tabs and the change in the consistency and thickness of the tabs. This is true that approximately 6-8 months ago the raw material that had been used over many years to make these tabs become obsolete. The manufacturers sourced these materials from the same manufacturers of type writer ribbon however when we all stopped using ribbon we lost the source for the carbon raw. The new material is a bit thicker than and not as sticky as the original but for most it is fine and works adequately. Alternatively we here at Electron Microscopy Sciences has found a new source from the Far East that has raw material very similar to the original which produces very thin tabs with the same sticky power as the old original. The problem is the new raw material is double the cost of what was. We do have ample stock of it for those of you whom are looking for the thin material. The good news is doing a search out there on all of the thin material our price of $60 per 200 tabs seems to be very low. Please do not hesitate to contact me if you have any questions I look forward to hearing from you. Sincerely,
Stacie Kirsch Electron Microscopy Sciences 215-646-1566
TECHNICAL DIRECTOR FOR CRYOELECTRON MICROSCOPY NEW YORK STRUCTURAL BIOLOGY CENTER
The New York Structural Biology Center (http://www.nysbc.org) seeks a Technical Director for a high-end, state of the art facility in Cryoelectron Microscopy. The facility will include three cryoelectron microscopes at 200, and 300 kV as well as ancillary equipment required for the three main cryoEM applications: tomography, single-particles, and crystallography. The facility will be used by in-house research groups and by investigators from a consortium of ten New York academic research institutions on a broad range of biological targets.
The appointee will initially be responsible for developing specifications, testing and installing the three instruments, and later for implementing specialized technologies such as automated data collection, for supporting users and for supervising a technical staff. A strong technical background in electron microscopy is essential and familiarity with biological samples or image analysis would be a bonus. Send a biographical sketch, a statement of research experience, together with names and addresses of three references to CryoEM Search Committee, at nysbc-at-nysbc.org.
-- ------------------------------ David L. Stokes Skirball Institute NYU Medical Center 540 First Ave New York, NY 10016 tel and FAX: 212-263-1580 http://saturn.med.nyu.edu/~stokes ------------------------------
At one time I needed large Mylar sheets for HV insulation around an X-ray tube. I found "drafting fim" (mylar) in an office supply store. In "the old days" draftsmen would use their Leroy and Rapidograph pens to make their large drawings on clear film for better reproduction (originally acetate, then Mylar). In these days of AutoCAD, the film might be a bit more difficult to find.
By the way, "Mylar" is a tradename for a polyester film not a polyethylene film. Kodak film (SO-163 and 4489) uses a Mylar base.
From a DuPont web page... Mylar® is an extraordinarily strong polyester film that grew out of the development of Dacron® in the early 1950s. During the 1960s cellophane gave way steadily to Mylar® with its superior strength, heat resistance, and excellent insulating properties. The unique qualities of Mylar® made new consumer markets in magnetic audio and video tape, capacitor dielectrics, packaging and batteries possible. By the 1970s, Mylar® had become DuPont’s best-selling film, despite mounting competition. Mylar® is now a product of a joint venture, DuPont Teijin Films.
Cheers, Henk Colijn
At 12:57 PM 5/28/2003 -0400, Geoff McAuliffe wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
Does anyone have a technique to visualize double stranded RNA molecules by TEM? Which way is one the best (negative staining, shadoving or combination...)?
Valentin Starovoytov The State University of NJ Division of Life Sciences Electron Imaging Facility
At 12:57 PM 5/28/2003 -0400, Geoff McAuliffe wrote: } } I am looking for some Mylar (polyethylene I think) sheets, clear not } 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any } ideas who/where sells this stuff?
Try Goodfellow Materials (www.goodfellow.com online catalogue) - they have mylar sheet in thicknesses 1,2,5 & 10 mm, 150, 300 and 600mm square.
-- Dr. Norman Charnley Department of Earth Sciences University of Oxford Oxford OX1 3PR, UK.
I asked on of my associates that deals with plastics and films. Here is his reply. Hope this helps.
..There are so many different grades of Mylar, for detailed product information and specifications for Mylar Polyester Film, try the DuPont-Teijin Film website at http://www.dupontteijinfilms.com/cntProductsMylar.htm
Also, you may call Shawn Ford, sales rep. of Plastic Films Co. at 1-800-445-0719 (I'm not sure if he is still there).
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, May 28, 2003 12:58 PM To: Microscopy
Dear List:
I am looking for some Mylar (polyethylene I think) sheets, clear not 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any ideas who/where sells this stuff?
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Just regular, old fashioned polarized light would seem to be the most direct solution. Even if the particles are small, they should respond as bright against the dark background of crossed polars. Based on the size, I would suggest higher mags (60x or even 100x). I would also make sure that you do not use plan apos but use objectives especially suited for polarized light analysis. To test if the system itself might cause a problem, put a sample in place, establish Koehler illumination, then cross the polars to get the blackest background possible. Remove the sample and see if the background stays velvety black. If you do not get velvety black at all steps, the optics are exhibiting strain and will interfer with your imaging. You may also want to inquire about the lab-level microscopes that are built specifically for polarized light analysis. Leica's have been well known for decades and Nikon came out with a new series about 3 years ago.
Re: the size of your particles The good thing about "bright against dark background" types of imaging is that it is not resolution limited: you will be able to detect the particles, even at 100nm. You may not be able to size them at that level, or to get good edge or shape information, but you should be able to detect that they are. Of course, the higher the NA on the objective (and condenser), the better the imaging in general. Oil immersion may not be a bad idea. I had my hands on a 40x/1.3NA two weeks ago which was incredible!
Re: sample prep. You may want to dilute the blood with an isotonic solution, just to avoid overcrowding from red blood cells, but unless the red blood cells are just too thick, I don't see any need to sonicate or do anything else as drastic as you have described.
Let me know how this works!
Best regards, Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration/ Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- At 09:40 PM 5/27/03 -0500, Damian Neuberger wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I want to model a surface one-dimensionally with respect to surface roughness. I would like to know whether I can take a segmented line where the nodes are displaced vertically from a horizontal line to approximate the RMS surface roughness and then simply take the standard deviation to give the RMS value. In effect, the segmented line would be the same as a line profile across the surface.
Secondly, what I would like to do is have a particular RMS value and randomly distribute the nodes such that the line profile would give me that RMS value.
If there is anyone that can comment on the validity of this approach or if there is already a method or model available, I would greatly appreciate it.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Does anyone have a procedure for preparing a sample to detect carbon using an electron microprobe? The particular samples are 50-100 um catalyst particles. Thanks for any help.
Greetings, I 'inherited' some old (well 1950's to 1970's) Leitz microscopes and accessories. The most interesting is an Ortholux 1 (?) with a complete fluorescence attachment that predates epi. That is, the light path is transmitted. It has an arc lamp of some kind (not carbon!) and sliding filters. There is also a box with all sorts of 'accessories', and another Lietz stand. This would be a great source of parts for collectors and afficionados. I don't want to store it any more. If anyone is interested in acquiring some or all of it, please contact me.
You mention a procedure to detect carbon, that's not so hard with a STE crystal. You should be able to make relative comparisons between your specimens. However, quantifying is another matter. I assume you be mounting and polishing the particles?
Re:carbon baseline in my probe. Due to carbin in pumping system (rp's and diff pump) I measure carbon in all samples if I leave beam on one spot long enough. To get around this I understand some have successfully used Bastin & Heijligers work. Check out their paper, "Quantitative Probe Microanalysis of Ultralight Elements (Boron-Oxygen), Scanning, Vol 12, 225-236 (1990). The authors introduce the idea of area peak factors APF. I've managed to avoid doing carbon work but I'll use APF's when it happens.
Owen
At 01:42 PM 5/29/2003 -0400, Katie.Gibbs-at-grace.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer 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
Mylar® is an oriented polyester (PET) film. The DuPont website below has info and a contact phone number in case you can't find a supplier. http://www.dupontteijinfilms.com/datasheets/mylar/overview/h67160.pdf You might also want to note that there are a variety of Mylar® options depending on your end use: http://www.dupontteijinfilms.com/FilmEnterprise/ProductLocatorResults.asp?ProductID=5&Version=US
As for who sells it, a quick "Google" search says everyone from graphics supply stores: http://www.grafixplastics.com/mylar.html to sailboat supply stores: http://store.catsailor.com/tek9.asp?pg=products&specific=jrrmdocrq (20"x27"x3mils) The catch is matching the exact characteristics and quantities you need with a possible source. Good luck with your search!
- Louise
Louise Harner Research Microscopist Albany International Research Co. 777 West Street, P.O. Box 9114 Mansfield, MA 02048 508-337-9529 phone 508-339-4996 fax Louise_Harner-at-albint.com
-----Original Message----- } From: Geoff McAuliffe [mailto:mcauliff-at-umdnj.edu] Sent: Wednesday, May 28, 2003 12:58 PM To: Microscopy
Dear List:
I am looking for some Mylar (polyethylene I think) sheets, clear not 'metalized', 2 mils thick should do it, 8x10 inches or a bit larger. Any ideas who/where sells this stuff?
Geoff
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
The Canon G2 has a much larger lens which contributes to the vignetting problem they are having. The Nikon 995 has a very small lens, in fact, only a 28mm threaded opening. The G2 has an external lens assembly that extends out of the front of the camera upon powering up. A lens "tube" is needed to compensate for this. Therefore, the lens of the camera and the upper element of the microscope are kept farther apart. The 995 has an internal lens assembly, which requires only a step ring and allows the camera lens to be much closer. The 995 is a much better choice for imaging through a microscope or a telescope.
A MaxView Plus adapter from klughammer gmbh system will "help" reduce the vignetting in the G2, but it will not eliminate it all together. Hope this helps!
For more information please contact me at schmaus-at-klughammer.de.
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bwoAAM} Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (rene.oberli-at-strasbourg.gm.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, May bwoAAM} 27, 2003 at 09:15:51 bwoAAM} ---------------------------------------------------------------------------
bwoAAM} Organization: process enginer at general motors strasbourg france
bwoAAM} Education: 9-12th Grade High School
bwoAAM} Location: strasbourg France
bwoAAM} Question: I would like to make digital pictures with Canon G2 photo camera thru a WILD M3 (6 to 40x power, no zoom 1X front lens)with a LEiCA trinocular head up to now I was not able to get a bwoAAM} full frame picture I also tried with special lens ( 0,6x )from LEICA or NIKON between the output tube and the G2 without success; the sharp image is circular and not full frame.This assembly bwoAAM} works fine with a NIKON coolpix 995.Why this dont work with CANON G2 with or without zooming the G2? Canon-Europe is no help from a contact with them they dont know or dont want to help!
At 08:02 AM 5/29/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
One of the scientists here has been asked by the local zoo for a method to preserve bacterial/fungal cultures for demonstration purposes. They are referring to culture plates, not microscope slides. The plates will probably be handled by many people so they can't be made of glass. Will some epoxy embedding chemical work or other such material?
Thanks for your assistance.
Bruce F. Ingber, Biologist Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124 bingber-at-srrc.ars.usda.gov 504-286-4270 phone 504-286-4419 fax
These are supposedly the thin ones. If they work out, they are cheaper than Fullam's units. If not right, then Fullam's tabs at $90/200 is the next US-sourced option I suppose.
I should have these thin ones next week. will advise how they work.
gary g.
At 05:30 AM 5/29/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I am presently working on reconfiguring an ISI DS 130C SEM to perform electron beam lithography. This involves using a high-definition 3 channel arbitrary waveform generator to power the X and Y scan coils and also the blanking plates to deflect the beam on and off the sample at high frequencies. The hardware setup required is in place now but we have encountered an issue.
The problem I am having presently is that our DS 130C SEM has been unused for several months and was moved to two different rooms and therefore (inevitably) the schematics have gone missing. External inputs to the scanning coils are normally given through a 14 pin terminal on the back labeled 'PGT' but the pin configuration of this input terminal is unknown. Finding literature that talks about this unit has also been a problem due to age issues.
I was hoping that someone could help me out on this by providing a schematic or pinout for that terminal. The connector at this terminal is a miniature ribbon style 14 pin metal shell bail-latching female connector.
The service contract for this machine is operational but our service guys don't do too many DS 130s and they keep all schematics at the customer sites, so our copy was one of the few they have access to.
Thanks a lot for any assistance possible. I can be reached at: chety-at-asu.edu
Chetan Prasad ------------------------------------ Research Associate Nanostructures Research Group Department of Electrical Engineering Arizona State University Tempe, AZ 85287-5706 Phn: 480-965-4097 Fax: 480-965-8058 ------------------------------------
Thanks to all who have sent me ideas to try with this project. They are all good and will probably end up trying them all in one fashion or another or in combination. I must say that this listserver is a tremendous resource especially when time constraints are very tight.
Thanks again and still looking for any other ideas that come across.
DEAR FRIEND, I AM OBLIGED TO CONTACT YOU FOLLOWING INFORMATION I GOT ABOUT YOUR COUNTRY. I WISH TO GO INTO BUSINESS RELATIONSHIP WITH YOU. I AM THE ONLY SON OF LATE MR. AND MRS.AKONAM MY MOTHER DIED IN 1986 WHEN I WAS FOUR YEARS OLD. THEN, MY LATE FATHER TOOK ME SO DEAR BECAUSE I WAS THE ONLY CHILD. MY LATE FATHER WAS A POLITICIAN, AN ACCOMPLISHED, WIDELY KNOWN AND RECOGNISED MILLIONAIRE FARMER IN THIS PART OF THE REGION. HE WAS ONE OF THE GREATEST SUPPLIERS OF CATTLE, BEEF AND OTHER DIARY PRODUCTS IN THIS PART OF THE COAST. UNFORTUNATELY, HE WAS MURDERED LAST YEAR BY HIS BUSINESS ASSOCIATES WHILE ATTENDING BUSINESS SEMINAR IN BOUAKE. BEFORE HE FINALLY DIED IN THE HOSPITAL, HE CALLED ME AND INFORMED ME ABOUT SOME WEALTH HE LEFT IN THE PRIVATE VAULT OF A BANK HERE IN COTE D’IVOIRE IN FORM OF A BARE BOND AND THAT HE USED MY NAME AS THE ONLY CHILD FOR THE NEXT OF KIN TO DEPOSIT THE MONEY. MY LATE FATHER EXPLAINED TO ME THAT HE TOLD THE BANK THAT HIS FOREIGN BUSINESS ASSOCIATE OWNED THE MONEY. HE ALSO TOLD ME THAT THIS MONEY WAS THE CAUSE OF HIS DEATH. HE ADVISED ME IF HE DIES THAT I SHOULD MAKE URGENT ARRANGEMENT TO TRANSFER THE MONEY TO A COUNTRY WHERE I LIKE AND USE THE MONEY FOR INVESTMENT. HE ALSO ADVISED ME TO LEAVE COTE D’IVOIRE AND RUN FOR MY LIFE TO PREVENT HIS ENEMIES COMING FOR ME. PLEASE IT IS ON THIS PREMISE I WRITE YOU. I AM CONTACTING YOU BECAUSE I WANT YOU TO CONFIRM TO ME ASAP YOUR POSSIBILITY OF PRESENTING YOURSELF TO THE BANK AS MY FATHER’S BUSINESS ASSOCIATE AND CLAIM THE MONEY ON MY BEHALF SINCE HE HAD DECLARED THAT THE MONEY BELONGED TO HIS FOREIGN BUSINESS PARTNER AND AS A BARE BOND, THE MONEY HAS NO SPECIFIED BENEFICIARY, HENCE ANY FOREIGNER CAN PRESENT HIM/HERSELF AS THE BENEFICIARY PROVIDED YOU PRESENT ALL THE DOCUMENTS ISSUED AS AT THE TIME OF DEPOSIT, WHICH I WILL FORWARD TO YOU. YOU SHALL ALSO BE REQUIRED TO ASSIST ME IN INVESTMENTS IN YOUR COUNTRY. I HAVE PLANS TO INVEST THIS MONEY ON REAL ESTATE AND INDUSTRIAL PRODUCTION. I WILL ALSO WANT YOU TO PROVIDE A BANK ACCOUNT WHERE THE MONEY WILL BE TRANSFERED TO BESIDES, YOU WILL BE MY GUARDIAN, TRUSTEE AND THE CUSTODIAN OF THIS MONEY. I HAVE NO KNOWLEDGE ABOUT INVESTMENT. I AM A MAN OF 20 YEARS. YOU WILL AGAIN HELP ME COME OVER TO YOUR COUNTRY WHERE I PLAN TO RESIDE AND FINISH MY EDUCATION ALSO. I TRUST YOU AS A GOD FEARING PERSON WHO WILL NOT SIT ON THIS MONEY WHEN YOU CLAIM IT, RATHER ASSIST ME PROPERLY. I PROMISE TO GIVE YOU 20% OF THE TOTAL MONEY FOR HELPING ME. THIS WE WILL WRITE AN AGREEMENT. I SPOKE TO THE BANK MANAGER YERSTERDAY AND HE TOLD ME THAT THE BANK WILL CONCLUDE THE TRANSFER OF THE MONEY TO YOUR ACCOUNT WITHIN 9 BANK WORKING DAYS AS SOON AS YOU CONTACT THEM AS THE OWNER OF THE MONEY. I WILL LET YOU KNOW THE NAME OF THE BANK AND SEND YOU THE DOCUMENTS FOR THE DEPOSIT AS SOON AS I HEAR FROM YOU PLEASE INDICATE YOUR INTEREST TO ASSIST ME BY SENDING YOUR PHONE AND FAX NUMBERS FOR IMMEDIATE CONTACT. THANKS AND GOD BLESS YOU. NKEM AKONAM.
-- Ziskejte kvalitu, kterou si zasluhujete. Za minimalni mesicni poplatek vam nabizime Antivir, Antispam nebo dalsi kapacitu pro vas Mailbox. Vice na: http://sluzby.volny.cz/product/postpaid/
Dear Halina, Absolutely no problem, You can store tissue fixed for resin embedding at 4 deg and for cryosectioning at RT.
Shashi
I would greatly appreciate your help in dealing with the following problem. We have a set of samples from an experiment on rats performed yesterday (Tuesday, May 27). The lung and aorta samples were divided into two parts each: one, for embedding in EMbed 12, was brought to the step of washing after osmium tetroxide staining and the other, for cryosectioning, was brought to the step of infiltration with PVP and sucrose. The first set of samples is currently in 0.1 M sodium cacodylate buffer, pH 7.4, and the other in the PVP with sucrose in phosphate buffer pH 7.0. However, there is a delay in delivery of the embedding kit and aluminum specimen pins. Most likely, both will arrive here on Monday, June 2nd. Is it safe to keep the samples at this stage of preparation until Monday? If yes, at what temperature?
Thank you, very much.
Sincerely,
Halina Witkiewicz, PhD
Senior Research Scientist
Division of Vascular Biology and Angiogenesis
SIDNEY KIMMEL CANCER CENTER
11835 Altman Row
San Diego, CA 92121
(858) 450 5990 x 326
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
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I have been asked to process adult mouse bone, a new tissue for me. I would appreciate any protocols or suggestions for processing this tissue from fixation through TEM plastic embedding, including decalcification steps. Also, should I be concerned about damage to my diamond knife during thin sectioning? Thanks in advance. Marilyn Levy
You may use both methods for visualization of dsRNA. Negative staining is a little tricky. Shadowing works better. You can use Kleinschmidt's method of spreading dsRNA with cytochrome c on buffer (water) surface or direct mounting method using spermidin buffer and addsorbtion on to glow charged carbon supports.
If you have more questions, please, fill free to email me.
Good luck! Udachi!
Alexander Makhov
Lineberger Comprehensive Cancer Center University of North Carolina at Chapel Hill Chapel Hill, NC 27599
----- Original Message ----- } From: "Starovoytov" {Starovoytov-at-nel-exchange.rutgers.edu}
} I was hoping that someone could help me out on this by providing a schematic } or pinout for that terminal. The connector at this terminal is a miniature } ribbon style 14 pin metal shell bail-latching female connector. } } The service contract for this machine is operational but our service guys } don't do too many DS 130s and they keep all schematics at the customer sites, } so our copy was one of the few they have access to. }
ISI DS130, J44 connector aka. "PGT" or "EXT" . Centronics Style (Amphenol 57-30140). External Scan Control Connector.
Pin 1 (Input) External Scan Control ( 0v internal, +5 to +12v external). Pin 2 (Input) External Brightness Control (+5 internal, 0v external, TTL). Pin 3 (Input) External Horiz Ramp ( +5 to -5 volts). Pin 4 (Input) External Vert Ramp ( +4 to -4 volts). Pin 5 not used. Pin 6 (Input) External Brightness (+14v (black) to -14v (white). Pin 7 (Input) External Blanking (+5 blanked, 0v unblanked, TTL). Pin 8-14 (Gnd) 0v.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 ext 13 (tel) 4pi Analysis, Inc. Fax: 919 489-1487 (fax) 3500 Westgate Drive, Suite 403 email: davilla-at-4pi.com Durham, North Carolina 27707-2534 web: http://www.4pi.com
What level of carbon? If it is greater than 1%, than embedded polished particles will do fine. You will not want to use sticky tape or carbon paint to mount your particles. For low carbon content you should use liquid nitrogen trap, and the trap is essential for quantification.
If your particles are not conductive, you can coat them with thin layer of C or Au, but if you coat with C than the reference point ('zero') will be intensity from substrate.
Good luck,
Vladimir
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} } Does anyone have a procedure for preparing a sample to detect carbon } using an electron microprobe? The particular samples are 50-100 um } catalyst particles. Thanks for any help. } } } Katie Gibbs } } e-mail: katie.gibbs-at-grace.com } } }
If you're forced to store half-processed tissue for TEM, it's better to keep it at the buffer stage after glutaraldehyde and before OsO4. Since it's too late for this, you might find it a good idea to repeat the osmium step when your materials arrive and you can continue. Meanwhile, keep it at +4oC, as Sergey says.
Lesley Weston.
on 29/05/2003 12:58 AM, Sergey Ryazantsev at sryazant-at-ucla.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I don't see big problem. Keep it refrigerated +4oC. Extended wash in } cacodylate may wash out some components of the cell. You may expect some } artefacts from that. If your samples for cryo-EM were fixed with } formaldehyde, you know: formaldehyde fixation is reversible. So, extended } wash after formaldehyde may cause "less" fixation. Sergey } } At 06:13 PM 5/28/2003, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I would greatly appreciate your help in dealing with the following } } problem. We have a set of samples from an experiment on rats performed } } yesterday (Tuesday, May 27). The lung and aorta samples were divided into } } two parts each: one, for embedding in EMbed 12, was brought to the step of } } washing after osmium tetroxide staining and the other, for cryosectioning, } } was brought to the step of infiltration with PVP and sucrose. The first } } set of samples is currently in 0.1 M sodium cacodylate buffer, pH 7.4, and } } the other in the PVP with sucrose in phosphate buffer pH 7.0. However, } } there is a delay in delivery of the embedding kit and aluminum specimen } } pins. Most likely, both will arrive here on Monday, June 2nd. Is it safe } } to keep the samples at this stage of preparation until Monday? If yes, at } } what temperature? } } } } } } } } Thank you, very much. } } } } } } } } Sincerely, } } } } } } } } Halina Witkiewicz, PhD } } } } Senior Research Scientist } } } } Division of Vascular Biology and Angiogenesis } } } } SIDNEY KIMMEL CANCER CENTER } } } } 11835 Altman Row } } } } San Diego, CA 92121 } } } } (858) 450 5990 x 326 } } } } HWitkiewicz-at-SKCC.org } } } } } } _____________________________________ } } Sergey Ryazantsev Ph. D. } Electron Microscopy } UCLA School of Medicine } Department of Biological Chemistry } Box 951737 } Los Angeles, CA 90095-1737 } } Phone: (310) 825-1144 } FAX (departmental): (310) 206-5272 } mailto:sryazant-at-ucla.edu } } } }
I'd suggest you contact one of the interferometry companies to validate this method. My customer support contact at Zygo is John Roth: 860-347-8506. My old contacts at Wyko (now Veeco, in Tucson) are gone, but I would also suggest that you give them a call.
Caveat: MME has no financial interest in this issue.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
Will you be at M&M in San Antonio? If so, don't forget the Tuesday night seminar on Fluorescence Calibration/ Also, join the tradition of over 10,000 microscopists: participate in our survey at any time during the meeting and receive a "sweet thank you". Booth #218 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
At 01:54 PM 5/29/03 -0400, Walck, Scott D. wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have adapted a Nikon Coolpix 5000 to our microscope with trinocular head. The adaptation is not specific to the microscope, and we are having a problem with focus. That is, we can elevate the stage for vision's focus, or we can elevate the stage for the camera's focus ... but neither are coincident ... and we imagine this is because the adaptation is not specific to the microscope (the adapter being quite expensive for this particular brand & model). Judging from playing with the adapter, its tube-length is slightly too long.
I suppose we were hoping the camera could be manually focussed, so that both could be made coincident, but for the Coolpix we cannot find a way of manually focussing. Or, it may be impossible to focus because the adapter's tube-length is too long rather than too short. Is this the correct assumption? What have others done?
In a message dated 5/30/2003 11:14:55 AM Eastern Standard Time, michael-at-shaffer.net writes:
} We have adapted a Nikon Coolpix 5000 to our microscope with trinocular } head. The adaptation is not specific to the microscope, } and we are having a } problem with focus.
You don't mention how you adapted the camera to the scope and whether there are optical elements in your adapter, but it sounds like you just have a straight tube with no optics in it.
For the Nikon cameras (and all other consumer/prosumer cameras with non-removable lenses), you need intermediate optics to form an image plane that can be projected through the camera's lens and onto the CCD.
The easiest way to do this is to use an eyepiece of some kind. You can verify this by holding the camera in front of one of the scope's viewing eyepieces, and you'll see that it makes a nice image through the camera.
If your adapter is homemade, you may have to modify it so that it will hold either an eyepiece or a photoeyepiece in the trinocular port. You'll also need to get the front of the camera's lens as close as possible to the eyepiece. Set the camera at "infinity" and don't use the autofocus, and you should be able to get fairly good images.
Note also, if you see a lot of vignetting (clipping of the image at the corners) you may have to use a little of the camera's digital zoom to completely fill the field of view.
Biophotonics International magazine is planning to coordinate an article that is a practical guide for research-grade light microscopy users (brightfield, epifluorescence, confocal and multiphoton). We would like your input on what is important to include in this article. This could be information that you teach in a microscopy course, common problems faced in your lab or a core facility, or perhaps issues that constantly frustrate you as a user.
If you would like to participate in this exchange please e-mail your questions directly to biophotonics-at-laurin.com with "Microscopy Guide" in the subject line. If you have suggestions on the best person to contact (company or researcher) for more information on the subjects you think are important, please include that in the e-mail as well. We may also contact you for more information on the subject.
If you are not familiar with the magazine, we cover the latest research related to biological applications of light-based techniques including microscopy. We will not print anything directly from the e-mail without first requesting your permission. For more information on the magazine or to sign up for a free subscription, please visit www.photonics.com/bio.
Sincerely, Nancy Lamontagne Senior Editor Biophotonics International
When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids, I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then?
Thanks in advance for any comments.
Ling
------------------- Ling Pan, Ph.D. 248 MRL Building Materials Research Institute The Pennsylvania State University University Park, PA 16802 Tel: (814)865-6115 Fax: (814)865-2326
I agree with Lesley: ideally, it's better to break procedure at the after GA fixation and store in the buffer refrigerated. Sergey
At 07:23 AM 5/30/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
We have been discussing the cryogens normally used for plunge freezing - things like propane and ethane and (formerly) Freons of various stripe. All have some rather significant downside, like potential environmental damage with Freons and the potentially explosive condensation of liquid oxygen in the hydrocarbons. This makes a rather routine and quite useful protocol fraught with drama, especially when it must be done in underground labs.
Do any of you materials folks out there know of a compound or class of compounds that have a rather large temperature spread between the freezing and boiling points and which remain liquid at or near liquid nitrogen temperatures (-196C) and which are also essentially inert with regard to oxidation potential and which (as long as I'm dreaming) have no real effect on human health? We were thinking that probably a whole bunch of stuff has been synthesized in the last 20 or so years that fit the bill (that is, since the pioneers of cryofixation did their initial search for likely fluids), but that the people who know about these compounds don't know why we would want to use them, and those of us who need new, safe cryogens don't really know about the possibility of these new substances.
Any leads or hints (or outright answers) are welcome. Thanks in advance for advice.
William P. Sharp Arizona State University Dept. Plant Biology, box 871601 Tempe, AZ 85287-1601 Phone - (480)-965-3210 Fax - (480)-965-6899
Ling, The Cu peaks may not be generated from the sample grid, but from the TEM itself. Backscattered electrons can cause x-rays from within the column. Try varying parameters such as accelerating voltage, detector position, spot size, etc. If that doesn't help, maybe contact your EDS and/or TEM manufacturer. There may be some things they can do to help to shield the detector from the Cu x-rays. Bill
Bill Carmichael Electron Microscopy Instructor Madison Area Technical College 3550 Anderson St. Madison, WI 53704
-----Original Message----- } From: LING PAN [mailto:lup2-at-psu.edu] Sent: Friday, May 30, 2003 3:09 PM To: Microscopy-at-sparc5.microscopy.com
When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids, I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then?
Thanks in advance for any comments.
Ling
------------------- Ling Pan, Ph.D. 248 MRL Building Materials Research Institute The Pennsylvania State University University Park, PA 16802 Tel: (814)865-6115 Fax: (814)865-2326
The camera can be set to manually focus, but that will not help your problem. You need to adjust the adapter to be in focus when the microscope binoculars are, although this is just for ease of use. You should focus the image using the camera before taking pictures and a separate larger monitor will help this. In general the little monitor on the camera is too small and cumbersome to use to focus with. Here is a link that may be helpful. A search on the web will turn up several other useful sites with good information...
http://www.microdapt.com/
----- Original Message ----- } From: "michael shaffer" {michael-at-shaffer.net} To: "Microscopy list" {Microscopy-at-sparc5.microscopy.com} Sent: Friday, May 30, 2003 9:26 AM
You will always see the Cu peaks with Cu grids in a TEM. This problem is well documented and discussed in several reference texts. I suggest that you find the Williams and Carter book on TEM for Materials Science. Nestor Zaluzec also wrote an excellent review article for MSA Bulletin a number of years ago and he has a presentation based on that for XEDS analysis in the AEM. I know this because he once directed me to his presentation and I have it somewhere. Unfortunately, I couldn't find it in the archives. I suggest that you contact him directly to see what he has.
The origin of the peaks are due to hard X-rays from the apertures coming down the column and striking your sample and from X-rays generated within your sample fluorescing the Cu grids. If you are not using an analytical sample holder, you will also get X-rays from that. You should also be using top hat apertures that are essentially thicker and help to reduce the hard x-rays. You will have to take what is known as a hole count and subtract that background signal from your data. This is done by putting the probe in a open area and collecting a spectrum. I also suggest getting rid of the Cu grids and using Be grids. You will not see a Be signal.
There is also a NiO test sample that can help you see how your system is working. Here is a web site that discusses it and talks a little about the spurious X-rays that you are seeing in your studies. http://www.tedpella.com/calibrat_html/TEM7.htm
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
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-----Original Message----- } From: LING PAN [mailto:lup2-at-psu.edu] Sent: Friday, May 30, 2003 5:09 PM To: Microscopy-at-sparc5.microscopy.com
When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids, I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then?
Thanks in advance for any comments.
Ling
------------------- Ling Pan, Ph.D. 248 MRL Building Materials Research Institute The Pennsylvania State University University Park, PA 16802 Tel: (814)865-6115 Fax: (814)865-2326
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In the previous answers regarding the CU peaks in STEM/EDS, the effects of the Cu grid and that of the hard X-ray coming out from the column have been explained. However not that of the backscattered electrons arising from the sample itself, which will hit the grid but more surely some parts of the specimen holder which frequently contains copper (just check that). The "heavier" the thin foil, the most pronounced the spurious Cu peak. Pt-Ru (heavy atomic species) are good candidates to enhance this artefact. It is very spectacular to 'sign' a 'heavy' precipitate within a 'light' matrix with the Cu peak with EDS maps. This effect cannot be explained by fluorescence arising from hard X-ray generated within the column of direct 'column photons' analyzed... Our experience comes from a JEOL 2010F. On the double-tilt specimen holder copper comes from the small 'cap' that is screwed on top of the sample. We did machine such a cap in titanium (which spares quite a lot of money compared with the solution to buy a Be-analytical holder...), and no more Cu peak (of course, a Ti one - but smaller - instead...)
Regards
Thierry EPICIER DR CNRS GEMPPM, umr CNRS 5510 Bât. B. Pascal INSA de Lyon F-69621 Villeurbanne Cedex Tel: +33 (0)4 72 43 84 94 Fax: +33 (0)4 72 43 88 30
First, has anyone noticed that the new tabs don't seem to be very conductive? I'm using Agar G3347 12mm conductive carbon tabs.
I've taken a multimeter to a tab on a stub and the resistance is off the scale. If I place a tab between two stubs and press them together as hard as possible, I still have a resistance of several megohms between the two Al stubs. Unfortunately I don't have any of the old, thinner ones to compare.
Second, has anybody else seen cases where the adhesive seems to have penetrated or collapsed a porous structure? We look at polymer films about 10-30 microns thick with holes ~100nm in diameter. On the thinner films especially, the bits of the film firmly in contact with the tabs are always less porous or non-porous than the bits that are suspended above. Could it be that the adhesive is that thick and that it's penetrating our films?
I am greatful to all of those who responded to my Query related to the problem I faced. I have alredy tried out one of the suggestions, and was able to get the desired blocks. I am planning on trying out the other suggestions too and work out on the one that gives me the best result.
Once again, Thank you for you timely assistance.
Joston P. Nongkynrih (Jos) Sophisticated Analytical Instrumentation Facility North-Eastern Hills University Shillong, Meghalaya India
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I know that there is concern in the community about whether what is being published as FRET is really FRET or not. I think we are seeing FRET but want to know where I could be misleading myself. We have expressed the same actin binding domain in cells from two different plasmids. One is a fusion with GFP and one with mRFP-1. The population is not uniform in expression, so that there are cells expressing green only, cells expressing red only and cells expressing both. Examining the cell population with a Leica SP2, we tuned the two channels to acquire green and red signals. The green channel is tuned to see green cells only when the 488 laser is on and under these conditions there is no signal in the red channel for those green only cells. With only the 547 laser on, we see the red only cells and nothing in the green channel. When both lasers are on simultaneously, we see green, red and yellow cells. Now when we image with the 488 laser only and look in the red channel, the cells that were expressing both show a signal that is similar to but not identical to either red or green alone, and the single expressing cells show nothing. As far as I understand things, this must be FRET. Am I missing any controls to verify this? Thanks-Dave --
Dr. David Knecht Department of Molecular and Cell Biology University of Connecticut 91 N. Eagleville Rd. U-3125 Storrs, CT 06269-3125 knecht-at-uconn.edu 860-486-2200 860-486-4331 (fax) home page: http://www.sp.uconn.edu/~mcbstaff/knecht/knecht.html
Dear Tamara- I am presuming the samples are fixed and sort of infilterated with sucrose-PVP,they are better left at RT. This is before they are cryofixed for sectioning. Shashi
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Ling Pan wrote: ============================================================================ = When I do STEM/EDS analyses on my TEM specimens on 200- or 300-mesh Cu grids , I always find Cu peaks present no matter where the beam shoots. Sometimes they can even be more intense than the peaks from my specimen when I am working on PtRu nanoparticles (~ 2 nm in size). Can anyone tell me the cause of the Cu signals? If the EDS spectrum shows signals from Cu grids which is more than a micron away from the beam, how can one be sure about the probe size or spatial resolution then? ============================================================================ = There are several "solutions" to this problem:
1] Use Be grids, see URL www.2spi.com/catalog/grids/beryl.shtml
and eliminate the copper lines. If you are working in an institution where Be is not permitted, we hope to have our diamond (made from diamond) mesh grids available again later this year, after having been temporarily taken out of production.
2] Silicon nitride membrane window TEM grids http://www.2spi.com/catalog/instruments/silicon-nitride.shtml
These grids, especially at the 30 nm thickness level, should be contributing very little to any extraneous secondary fluorescence effects. I won't say zero but the problem will be far less bothersome than with a copper grid, especially if you are doing your work near the center of the window.
Disclaimer: SPI Supplies has been a long time global supplier of both beryllium TEM grids and also, silicon nitride membrane window TEM grids.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
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by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id DAA04017 for dist-Microscopy; Mon, 2 Jun 2003 03:33:19 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id DAA04014 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Mon, 2 Jun 2003 03:32:48 -0500 (CDT) Received: from mail.iskon.hr (mail.iskon.hr [213.191.128.4]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id DAA04007 for {Microscopy-at-sparc5.Microscopy.com} ; Mon, 2 Jun 2003 03:32:35 -0500 (CDT) Message-Id: {200306020832.DAA04007-at-sparc5.microscopy.com} Received: (qmail 1649 invoked from network); 2 Jun 2003 10:26:21 +0200 Received: from ri02-037.dialin.iskon.hr (HELO ?192.168.1.100?) (213.202.65.38) by mail.iskon.hr with SMTP; 2 Jun 2003 10:26:21 +0200 To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Chris Blanford wrote: ============================================================================ ====== First, has anyone noticed that the new tabs don't seem to be very conductive? I'm using Agar G3347 12mm conductive carbon tabs.
I've taken a multimeter to a tab on a stub and the resistance is off the scale. If I place a tab between two stubs and press them together as hard as possible, I still have a resistance of several megohms between the two Al stubs. Unfortunately I don't have any of the old, thinner ones to compare.
Second, has anybody else seen cases where the adhesive seems to have penetrated or collapsed a porous structure? We look at polymer films about 10-30 microns thick with holes ~100nm in diameter. On the thinner films especially, the bits of the film firmly in contact with the tabs are always less porous or non-porous than the bits that are suspended above. Could it be that the adhesive is that thick and that it's penetrating our films? ============================================================================ ======= At SPI Supplies, over the years, we have been contacted by customers who have had one problem or another with the use of not only their double sided conductive carbon tabs (we call them discs), but also essentially the same product in the form of either "carbon tape" or "carbon sheets". As a side comment, it has always been somewhat of an amazement to me that so many users of these products seem to forget where they actually did make their purchase, and once we press for further information, it turns out that the product of their frustration did not even necessarily have an SPI Supplies progeny.
But one aspect of these kinds of products (all of them, irrespective of the form, or vendor) is that they do have a finite shelf-life, meaning specifically, that with the passing of time, there is a loss of properties, the most noticeable one being loss of tac. This is also accompanied with a loss of electrical properties (the material becomes more resistive). So if you are interested in extending the shelf life of your stock of such products, they should be stored refrigerated since this will slow down significantly the changes (storing at 10 C deg. lower will slow down the ageing rate by roughly a factor of ten times). A carbon disc that is past its prime, because of the higher resistivity, can even seem to be "bubbling" or experiencing other strange behavior. But this is not a matter of thin vs. thick, but one of age.
As an aside, for some, especially those using these products in UHV systems, the off-gassing characteristics are as important if not more so than the resistivity characteristics. There are indeed differences in the vacuum compatibility between these types of products from different sources. I will avoid making comment about where the SPI Supplies origin products stand in this respect, but only to make the point that we have arrived at a system that is optimized for a number of properties other than just electrical conductivity.
Unfortunately, for the typical user, unless they are making their purchase from the "ultimate source", and they have some reason to believe they are indeed receiving "fresh" material, the important variable of remaining shelf -life is introduced. We have seen instances over the years where a customer purchases material from a stocking distributor but unbeknown to them, the material had sat in that inventory for some number of years. So while it might be a "fresh" purchase so far as the customer is concerned, the material they are actually receiving is far from being fresh. Should the problems someone might be having be with a product of SPI Supplies origin, send us the lot number on the maroon SPI Supplies label and at least as a starting point, we can establish how old or fresh is that particular material.
We have ourselves directly received reports supposedly comparing the so- called thinner material vs. the so-called thicker material. We have not ourselves actually run controlled experiments side by side to compare the thin vs. thick material but only because we have not had any reason to do so . The SPI offered material has always been what one would call the "thick" material, and we are not aware of anyone who has really had problems with the SPI product. Naturally if one was having some kind of problem, we would like to know about it so that we could better understand the cause of such a problem.
But when the material eventually gets into the hands of the final end user, in addition to the potential variation in remaining shelf life, other differences can be introduced such as for the die cut discs (when the die cutting involved too much pressure), some of the silicone release liner can be transferred to the disc, at times so much so that the disc becomes non- conductive (from an SEM perspective) and it could also result in "bubbling". One way to test out if this is indeed a problem you are experiencing, compare the result you get with the disc vs. the result you get with carbon tape (making sure it is of the same material). If the carbon tape "works" but the disc does not, all other factors such as age being the same, then it would be our own prediction that the problem is the result of release liner transferral to the disc during the die cutting.
So my point is, it is clear that this could be much more than just "where it came from" or whether it is "thin" vs. "thick" material. If you have had favorable experience with the carbon tabs from a particular vendor, then you are getting fresh and not stale dated material. If you want to extend the useful lifetime of your carbon adhesive products, store them refrigerated. If you have had a less-than-satisfactory experience with your double sided conductive carbon products, and are looking to make a change, consider the range of products on URL http://www.2spi.com/catalog/spec_prep/cond_adhes.html
If you have a UHV system, consider the 6 mm wide tape product, make expressly to minimize the amount of organics put into the vacuum system.
Disclaimer: SPI Supplies was one of the first in the world to offer double sided conductive carbon products for SEM applications so we would naturally have a vested interest in calling our range of products to everyone's attention.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
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