I' m thinking to do double staining with peroxidase-DAB AND gold particles with silver enhancement in cultured cells (pre-embbeding treatment and I will use Epon). Do you know if it is possible? Do you have any reference about this technique? I will be very grateful for any information
Sincerely
Francisco Javier Hernandez-Blazquez IARC-WHO Unit of Multistage Carcinogenesis 150 Cours Albert Thomas 69372 - Lyon France fhernandez-at-iarc.fr Telephone: (33) 472738536 Fax: (33)472738442
is there any nice SEM pictures of beetles on WEB? I would like to create link from www.coleoptera.org to some nice image sites...
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
http://www.coleoptera.org Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., P.O.Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
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Two major international databases of entomological and agricultural literature are now available for free on WWW---
you can find both on www.coleoptera.org in directory {Coleoptera Bibliography} and in first subdirectory {Search}
Keep care and be of good cheer.
Regards
(name) Vratislav Richard Eugene Maria John Baptiste (surname) of Bejsak (Bayshark)-Collorado-Mansfeld
http://www.coleoptera.org Coleoptera - Australia, Tenebrionidae of World (incl. Lagriinae, Alleculinae)
University of Sydney The Wentworth Bldg., P.O.Box 62 NSW 2006 AUSTRALIA phone : +61 414 540 465 e-mail: vratislav-at-bigfoot.com ricardo-at-ans.com.au (before Ricardo-at-compuserve.com and ricardo-at-login.cz )
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.37 / Virus Database: 30 - Release Date: 6.4.1999
At 12:18 PM 4/30/99 , you wrote: } } Dear List, } } I would be interested in hearing about the value of service contracts. Our SEM } (Hitachi S-3100H) is about 2.5 years old and has been operating without } problems. If I operated without a service contract, handling all preventitive } maintenance myself, am I begging for trouble? Those of you who don't operate } with a service contract have you found the cost of emergency services so } expensive that a service contract would have been preferable? Do you find the } service difficult to get without a contract? Are the parts that would need } replacing terribly expensive (more than the cost of the service contract)? As } the instrument gets older is there a greater concern? Is my concern about the } cost of the service contract unwarranted? } } Thanks. } } David Rose } W.L. Gore & Associates } 297 Blue Ball Road } Elkton, MD 21921 } }
A good question and one that is difficult to answer. There are many possibilities.
It is true that as scopes age, as with any instrument, something can go terribly wrong. A pump can fail, lens coil can short out, driver blows up, CRT dies, etc. Depending on the particular scope and what type of source it uses, there is probably some crossover point where it makes sense to have the contract. For example, if you have a system running field emission, then the cost to replace the emitter is about $3500 just for the emitter. A LaB6 would run about $550 and W about $20. The cost of the annual contract would include the cost of replacing the emitter whether it actually needed replacement or not. If the emitter did not fail, you lost money. If it did, maybe you broke even. You probably just did not lose as much.
Suppose an ion pump fails or needs overhaul. This can cost between $500 and $1000. A mechanical pump rebuild/repair is about $350. Apertures are cheap and are simply consumables. The pumps will eventually need repair and overhaul. With normal use and reasonalble care, that is probably at the 4-6 year time point.
What I found works for me is to set aside about 1/4 of the annual maintenance contract cost in a separate account. I perform the PM myself. Then, if I need factory or independent service help, I have a pool to pay for it. And it does not hurt to have a factory or independent expert come in once in awhile just to check the system over and do some of the more exotic cleaning and alignment chores. These people generally charge straight time plus travel. some independents only charge for on-the-job time. The going rates seem to be about $125-$175 per hour. Most jobs tend to run around 4 hours.
If your system is really unreliable or you are not able to perform most of the PM functions yourself, then I would think that the maintenance contract is a good idea...either with the factory or an independent.
parts don't seem to be a problem. It is true that as systems get older it can be more difficult to obtain parts. However, this depends on the brand and what it is that needs replaced. The major items that need replacement and more or less industry standards. Like Edwards or Alcatel mechanical pumps. Varian ion pumps, etc. O-rings are widely available. It would seem that an item must be very scope specific to become an issue of availability. Within 5-7 years of a scope coming out, parts don't seem to be a problem. My early Amray is still supported by Amray.
We are seeking to replace our stock of these, and e-mail enquiries to the company themselves have got nowhere. So (a) does anyone know a supplier, and (b) is there an alternative? We have used this type because it is the only organic material for filter membranes that can stand up to Xylene at 120^C, and the anodisc alumina filters in our catalogues are all too small in pore size. =================================================== We (e.g. SPI Supplies) have offered the SPI Silver Membrane filters since 1976. They might be an acceptable alternative to the other mentioned choices. They are available in a number of different pore sizes including your desired 0.8 um. They are inert to xylene or just about any other organic solvent for that matter. You can get full information on our website given below. They handle more or less like other membrane filters including the fact they can be critical point dried.
While the expense per membrane is higher than most polymer membranes, depending on what you are collecting, if strictly organic, exposure to an oxygen plasma etcher can usually regenerate them for further use with other samples, something you could not do with polymeric membranes.
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: www.2spi.com ############################ ==================================================
we have to analyse by TEM magnetic samples coming from steel industry. The first tries we made in the TEM (LEO 912 Omega) were really not successfull, and we have no experience with this kind of samples.
Does anyone know how to handle magnetic samples in a TEM?
Many thanks in advance, Nathalie _________________________________________________
Dr. Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 e-mail : Nathalie.Bozzolo-at-crpcu.lu _________________________________________________
I truly appreciate all of you MSA list members taking the time to respond to my query about the old version of MSmouse.com driver. I was able to resurrect the old computer and discovered a bad RAM socket as main culprit. I will be able to copy the file from there to a new machine this week. Much appreciated and gracias!
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
have fun... sigh... I certainly have a lot more respect for the pioneers of EM who got all those nice photos of steel in the 50's.
Since the sample is likely to be a low carbon steel, you will probably find that the surface is covered with an oxide film which obscures any structure inside the sample. Even storing the samples in a vacuum desiccator did not prevent the formation of the oxide layers. Immediately before putting the sample in the scope, I have used a Fischione plasma cleaner running pure argon to clean this oxide layer off the surface. You may be able to briefly sputter in an ion mill to do the same.
One of the more frustrating aspects of doing EM on magnetic samples is that every time you move the sample (especially tilt), you will find it necessary to reset your current centering. I will often define one of the DF channels to be my bright-field condition so that I can easily reset my illumination tilt. (If readjusting the BF tilt is easy on your scope, this may not be an issue for you.) Otherwise, just remember that patience is a virtue.
Good luck, Henk
At 10:05 AM 5/3/99 +0200, you 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://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
I'm not quite sure if you got problems in sample prep or analysis 'cause magnetic material samples usually are not only tough to prepare but also tough to be analyzed. The following is just two tips:
1. Demagnetization This may work well only for hard magnets. Soft magnetic materials are easily magnetized anyway and therefore demagnetization wont help much. 2. ALWAYS turn off the objective lens while you are loading your specimen into the TEM.
As for analysis, that's at least a 3 credit 600 level course. You may want to see cells, walls, precipitates in hard magnets, or you may want to see anti-phase-boundaries in soft magnets and therefore superlattices, or most probably you may want to see magnetic domains and therefore you may even want to modify your TEM with special pole pieces, etc. etc. For functional multilayers, there are a bunch of other stories.
Good luck.
-cy
On Mon, 3 May 1999, Nathalie Bozzolo wrote:
} Date: Mon, 3 May 1999 10:05:53 +0200 (MET DST) } From: Nathalie Bozzolo {bozzolo-at-crpcu.lu} } To: Microscopy-at-sparc5.microscopy.com } Subject: TEM on magnetic samples } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear colleagues, } } we have to analyse by TEM magnetic samples coming from steel industry. } The first tries we made in the TEM (LEO 912 Omega) were really not successfull, } and we have no experience with this kind of samples. } } Does anyone know how to handle magnetic samples in a TEM? } } Many thanks in advance, Nathalie } _________________________________________________ } } Dr. Nathalie Bozzolo } Laboratoire d'Analyse des Materiaux } Centre de Recherche Public - Centre Universitaire } 162a, avenue de la Faiencerie } L-1511 Luxembourg } tel : (352)46 66 44 402 } fax : (352)46 66 44 400 } e-mail : Nathalie.Bozzolo-at-crpcu.lu } _________________________________________________ } } }
I work for a NASA contractor whose job it is to design and construct experimental packages for use in space. Currently we are designing a package which will make use of a Leica microscope to be flown aboard the International Space Station.
My question regards the use of immersion oil in conjunction with an objective. We have purchased an objective designed to be used as such, and it's operation is understandable on ground. What I need to know is if there is any one who has done any rersearch into the wetting properties of these types of oils, such that when we try to deploy them in space we can wet the surfaces of interest, i.e. the sample slide, and the objective? The necessity for this type of information becomes evident when you realize that there will be no gravity to assist in the deployment of the oil droplet.
Any assistance in the this matter would be greatly appreciated. Thank you in advance
Our lab owns a tem (philips 420, 15 years old) and a sem (nanolab le21000 = 12-15 years old) and we never had a service contract on any of them...raiso= n it is too expensive...
I believe that if you do basic maintenance such as oil change, filament = change...etc.... and you train and watch carefully people that are using = the instruments you should not run into deep trouble... I have compagny maintenance when I encounter a problem that I am unable to = solve and some of them are simply handle by the phone,=20 and since I have been working here (for the past 10 years), it cost me a = fraction of the cost of a contract service.... we now own a new SEM and intend to do exactly the same thing.
Hope this will help,=20
Diane Montpetit Electron microscopy lab Agriculture Canada Food research center St-Hyacinthe, Qu=E9bec Canada
Dear Aley, Because it is easy to slightly over-saturate the filament, I always run at just under the saturation point. Also, the saturation point actually creeps down as the filament ages, so check it, reset it and recentre it every hour for the first five hours or so of the new filament's life. The vacuum condition is also important. My filaments last an average of one month (100 hrs.) At 02:38 PM 02/05/99 +0300, you wrote: } Hello everyone, } } What is the average lifetime of a hair-pin tungsten filament? } We have a JEOL-840A SEM which we operate at 20 KV, and which we run at } asturation (on average) for ~ 7 hours everyday 5 days a week. For the } past several years, filaments used to last 6 weeks or longer, but } recently, the filament is burning out every 5 - 7 days! What could be } the reasons for this, and what are the solutions? } Many thanks. } } Aley El-Shazly } Department of Earth Sciences } College of Science } Sultan Qaboos University } POBox 36, Al-Khod PC 123 } Oman } e-mail: aley-at-squ.edu.om } 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-interchg.ubc.ca
} My question regards the use of immersion oil in conjunction with an } objective. We have purchased an objective designed to be used as such, and } it's operation is understandable on ground. What I need to know is if there } is any one who has done any rersearch into the wetting properties of these } types of oils, such that when we try to deploy them in space we can wet the } surfaces of interest, i.e. the sample slide, and the objective? The } necessity for this type of information becomes evident when you realize } that there will be no gravity to assist in the deployment of the oil droplet. }
Dear John, I have not studied the properties of immersion oil, but from what I've seen, the oil will wet both the glass of the slide and the parts of the lens. This property is a function of the composition of the oil and the microscope parts and will be independent of gravitation. That is, once the oil is in contact with the slide and the scope, it will form much the same contact angles as it does in an earth- bound lab. I suggest filling a syringe with the oil. On the space station, when examining a specimen on a slide place the lens near the slide. The distance should be somewhat smaller than the size of a droplet being expelled from the syringe. Place the syringe near the end of the lens and slowly express a drop of oil. When the drop makes contact with the slide and lens, it should wet both. Continue until the appropriate amount of oil is on the lens and slide. Good luck. Yours, Bill Tivol
Sounds interesting. I have no real experience in this matter but just a few things I thought about: Oil should work regarding stickiness but the surface tension may be a problem. Have you considered using the 63X/1.2 water immersion objective from Leica. I've heard it's really good and it will be less greasy:-) The surface tension of water will make it easy to control the droplet. If one equip the scope with both objective one can always see. I think there is a good 100X water objective from Leica as well.
Regards
Lars
____________________________________________________________ Lars Bjork,PhD
Dept of Immunology Dept. of Biology Wenner-Gren Institute Sect. for Cell & Mol. Biology Stockholm University Pharmacia & Upjohn S-106 91 Stockholm S-112 87 Stockholm SWEDEN SWEDEN
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Gentlemen, } } I work for a NASA contractor whose job it is to design and construct } experimental packages for use in space. Currently we are designing a } package which will make use of a Leica microscope to be flown aboard the } International Space Station. } } My question regards the use of immersion oil in conjunction with an } objective. We have purchased an objective designed to be used as such, and } it's operation is understandable on ground. What I need to know is if there } is any one who has done any rersearch into the wetting properties of these } types of oils, such that when we try to deploy them in space we can wet the } surfaces of interest, i.e. the sample slide, and the objective? The } necessity for this type of information becomes evident when you realize } that there will be no gravity to assist in the deployment of the oil droplet. } } Any assistance in the this matter would be greatly appreciated. } Thank you in advance } } John } John Eustace } Optical Physicist } Dynacs Engineering } 2001 Aerospace Parkway } Brookpark, Ohio 44142 } Ph (216) 977 - 1244 } Fax (216) 977 - 1269 } }
} } What is the average lifetime of a hair-pin tungsten filament? } } We have a JEOL-840A SEM which we operate at 20 KV, and which we run at } } asturation (on average) for ~ 7 hours everyday 5 days a week. For the } } past several years, filaments used to last 6 weeks or longer, but } } recently, the filament is burning out every 5 - 7 days! What could be } } the reasons for this, and what are the solutions? } } Many thanks.
Check the vacuum first. We had a long term problem with ~30 hours filament lifetime on a XL30/tmp. Tried all sorts of other fixes first. Replacing the gun o-ring fixed the problem and we are now up to ~160 hours. The gun o-ring is the cheapest fix, easy to do. Should have tried that first.
Scott ----------------------------------------------------------------------- Scott D. Davilla Phone: 919 489-1757 (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
We work on magnetic samples here and one of the primary methods we use to handle them is to reduce the volume of material as much as possible. This includes making the disks smaller than 3 mm (2.3 mm is another standard) and keeping them thin. I believe one technique has been to punch out a 1mm disk of the steel, and insert it into a 3mm disk of another material with a 1mm hole punched out. If you have questions about that technique, please contact me "offline" and I'll put you in touch with one of my colleagues. A more difficult sample prep method that has been around for some time is the "window" electropolishing technique. This method, outlined in many microscopy texts (Williams and Carter, for example), results in a small sample volume. It takes practice but can yield good results.
Additionally, we have ordered our microscopes with "extra strength" objective stigmator coils. If you will do many of these samples, perhaps they can be retrofitted (is that a word?) into your LEO.
Best of luck.
Cheers, JSV
*************************** John S. Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 USA Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
---------- } From: bozzolo-at-crpcu.lu Sent: Monday, May 3, 1999 12:05 AM To: Microscopy-at-Sparc5.Microscopy.Com
Dear colleagues,
we have to analyse by TEM magnetic samples coming from steel industry. The first tries we made in the TEM (LEO 912 Omega) were really not successfull, and we have no experience with this kind of samples.
Does anyone know how to handle magnetic samples in a TEM?
Many thanks in advance, Nathalie _________________________________________________
Dr. Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 e-mail : Nathalie.Bozzolo-at-crpcu.lu _________________________________________________
Couldn't you duck the issue and use high/dry objectives instead? These offer VERY good optics without the necessity for immersion oil.
Ann Lehman Trinity College Hartford CT
-----Original Message----- } From: John Eustace [mailto:John.Eustace-at-lerc.nasa.gov] Sent: Monday, May 03, 1999 9:15 AM To: Microscopy-at-Sparc5.Microscopy.Com
Gentlemen,
I work for a NASA contractor whose job it is to design and construct experimental packages for use in space. Currently we are designing a package which will make use of a Leica microscope to be flown aboard the International Space Station.
My question regards the use of immersion oil in conjunction with an objective. We have purchased an objective designed to be used as such, and it's operation is understandable on ground. What I need to know is if there is any one who has done any rersearch into the wetting properties of these types of oils, such that when we try to deploy them in space we can wet the surfaces of interest, i.e. the sample slide, and the objective? The necessity for this type of information becomes evident when you realize that there will be no gravity to assist in the deployment of the oil droplet.
Any assistance in the this matter would be greatly appreciated. Thank you in advance
The usual problem with the kind of samples you might get from a steel = company is that normal TEM-size disk samples made by jet electropolishing interact = so strongly with the objective lens (OL) magnetic field that they grossly = deflect the electron beam off axis and cause high image astigmatism. Moreover, = the beam axial alignment and astigmatism change each time the sample is tilted = (or sometimes even translated). In addition, thin foil areas tend strongly = to align normal to the lens axis, and will bend to maintain this alignment whan = you attempt to tilt the sample. What usually happens during tilting is, = just as you're approaching the orientation or diffraction condition you wanted, = the sample area reorients itself. This is a semi-reversable process, but = after you tilt back and forth a few times, the sample tends to become deformed = and worthless. In really severe cases of sample-OL field interaction, the = sample can be ripped out of the holder or will reorient the holder tilt = mechanism so the holder cannot be withdrawn without scratching the OL polepieces. = If this happens, don't try to remove the holder. Just turn off the OL, open = the OL chamber, and carefully free the stuck parts. In any case, you'll want = to make sure your sample is firmly held before inserting the sample holder. = The infamous 'j-ring' holder clips (c-shaped spring clips) in certain = holders are prone to losing magnetic samples and should be avoided. Screw-down = holder mechanisms are best. =20
OK. So what to do. Use the least amount of magnetic material you can = in the sample. Make the samples as thin as possible by reducing the sheet = thickness before punching disks for jet thinning. Disk samples 30-50 =B5m thick = can be used in most microscopes with foolproof sample mounting if you realign beam = tilts and correct OL astigmatism each time the sample is tilted. Many = microscopes increase the beam deflection range in the darkfield mode, so I = typically use one of the darkfield channels for magnetic sample work. Also, it's a good = idea to switch off the OL before inserting the sample holder. On some = microscopes, you can also increase the range of stigmator strength, but that's extreme. = Just remember to align the microscope beforehand with a nonmagnetic sample. = Also, your co-workers will appreciate it if you restore normal settings when = you finish. =20
In some cases, you might want to take the idea of minimizing magnetic = sample sizes further. A colleague working with radioactive samples has = developed the technique of punching out a 1 mm diameter disk sample and fastening it = into an annular disk of nonmagnetic material (usually 316 stainless steel) = before jet electropolishing the center. (I'll send the reference to you if I can = find it.) For much greater sample reduction, I have had success window-thinning = sheet samples (see standard references on TEM sample preparation for details) = and cutting off thin shards sandwich between two grids. In this case, = sample/OL interaction was almost nil. Good luck
Larry Thomas Mechanical and Materials Engineering Washington State University Pullman, WA 99352 USA
thomas-at-mme.wsu.edu tel: 509 372-0793 ---------- From: bozzolo-at-crpcu.lu Sent: Monday, May 3, 1999 1:05 AM To: Microscopy-at-Sparc5.Microscopy.Com Subject: TEM on magnetic samples
The Microscopy ListServer -- Sponsor: The Microscopy Society of = America=20 To Subscribe/Unsubscribe -- Send Email to = ListServer-at-MSA.Microscopy.Com On-Line Help = http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html = -----------------------------------------------------------------------.=
Dear colleagues,
we have to analyse by TEM magnetic samples coming from steel industry. =
The first tries we made in the TEM (LEO 912 Omega) were really not successfull,=20 and we have no experience with this kind of samples.
Does anyone know how to handle magnetic samples in a TEM?=20
Many thanks in advance, Nathalie _________________________________________________
Dr. Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 e-mail : Nathalie.Bozzolo-at-crpcu.lu _________________________________________________
Dear Sir/Madam, I am conducting experimental studies on surfactants. I would like to know if the temperature and concentration phase diagram of teepol is available. If it is please tell me where. Thanking You, Yours Sincerely, Geetha Basappa.
****************************************** Dr Geetha Basappa, Project Associate, Physics Department, Indian Institute of Science, Bangalore, India.
Aley, Check your upper & lower gun cylinder walls area for a faint bluish-gray tint(tungsten oxide). You could have a slight vacuum leak. You would probably never see the leak, it being so far away from any vacuum gauge. Good luck.
Gary M. Easton, Pres. Scanners Corporation ----- Original Message ----- } From: Mary Mager {mager-at-interchange.ubc.ca} To: aley {aley-at-squ.edu.om} Cc: {Microscopy-at-Sparc5.Microscopy.Com} Sent: Monday, May 03, 1999 11:39 AM
Mary Mager wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Aley, } Because it is easy to slightly over-saturate the filament, I always run at } just under the saturation point. Also, the saturation point actually creeps } down as the filament ages, so check it, reset it and recentre it every hour } for the first five hours or so of the new filament's life. The vacuum } condition is also important. My filaments last an average of one month (100 } hrs.) } At 02:38 PM 02/05/99 +0300, you wrote: } } Hello everyone, } } } } What is the average lifetime of a hair-pin tungsten filament? } } We have a JEOL-840A SEM which we operate at 20 KV, and which we run at } } asturation (on average) for ~ 7 hours everyday 5 days a week. For the } } past several years, filaments used to last 6 weeks or longer, but } } recently, the filament is burning out every 5 - 7 days! What could be } } the reasons for this, and what are the solutions? } } Many thanks. } } } } Aley El-Shazly } } Department of Earth Sciences } } College of Science } } Sultan Qaboos University } } POBox 36, Al-Khod PC 123 } } Oman } } e-mail: aley-at-squ.edu.om } } } 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-interchg.ubc.ca
Aley, With a sudden drop in filament life, as you describe, and assuming that you are treating your microscope the same as always, I would look for a vacuum leak in the immediate area of the gun. It could be a single lint fiber causing enough degradation of your vacuum to shorten your filament life as you descibe. Have you opened anything either on the back side of the gun (around thge stanpipe to the DP) or down on the column (fimal apertures, beam current probe, isolation valve)? If you have, that is the first place I would look.
Ken Converse owner Quality Images third party SEM service Delta, PA
Diane Montpetit wrote ... } } Our lab owns a tem (philips 420, 15 years old) and a sem } (nanolab le21000 12-15 years old) and we never had a service } contract on any of them...raison it is too expensive... } } ...
This is my viewpoint as well ... HOWEVER, I am a firm believer in finding the money (by whatever means possible) to cover a service contract for the two years after the initial warranty period. If anything is going to go wrong it will generally happen during this time period. This also gives the facility manager and technicians the time to become acquainted with the little idiosyncracies of the intrument with the help of those who are most aware.
... my $0.02 :o)
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Dear Listers, I am searching for an old JEOL 100SX TEM which could be used for parts. I would appreciate any leads. Thank you and please contact me at my address rather than on the list. Alex Greene SCIENTIFIC INSTRUMENTATION SERVICES, INC. PMB - 499 1807 West Slaughter Lane #200 Austin, Texas 78748-6200 Phone: 512/282-5507 Fax: 512/280-0702
At a recent MSA meeting (within the last 5 years or so) there was a session on microscopes in the classroom that featured something affectionately referred to as a 'Scope on a Rope'.
It was a video camera tethered to a TV with a built in light source and medium mag. All you did was hold it up to something and a magnified picture appeared on the TV.
Of course, now that I need to know more about it, I can't find any references. Anyone remember it or have an idea of where to start looking?
As always, thanks a million.
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Has anyone had any experience with phycoerythrin-conjugated antibodies and fluorescence microscopy? I'm specifically interested in hearing whether there are any "anti-fade" reagents that work well with PE. Although I've heard that PE antibodies don't work well with fluorescence microscopy, I don't have a choice of conjugates, unfortunately! Any help would be much appreciated!
Rizwan Haq Ontario Cancer Institute Toronto, Ontario Canada haqr-at-oci.utoronto.ca
Using Ultramicrotomy in Materials Science - September 21-24, 1999
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Lodging, tuition, meals, and materials are all covered in the fee of $1950 (only transportation excluded). Lectures and lodging are at the beautiful resort, Lodge on the Desert, below the Santa Catalina mountains. Lab sessions are at the RMC/Ventana's Tucson laboratory. For more information please contact Steve Miller, tel: 520-903-9366, SMiller-at-Ventanamed.com or see our web site at RMC-Scientific.com/microtomes/
} John Eustace wrote: } } } My question regards the use of immersion oil in conjunction with an } } objective. We have purchased an objective designed to be used as such, and } } it's operation is understandable on ground. What I need to know is if there } } is any one who has done any rersearch into the wetting properties of these } } types of oils, such that when we try to deploy them in space we can wet the } } surfaces of interest, i.e. the sample slide, and the objective? The } } necessity for this type of information becomes evident when you realize } } that there will be no gravity to assist in the deployment of the oil droplet. } } } John,
There is also a water immersion lens. It does not have as large an aperture as a oil immersion lens. But the water is a lot less noxious stuff to be floating around in the air than oil.
It won/t wet out as well as oil but spilled water is of little consequence in a weightless environment.
Water immersion should work for most experiments and you can always put the oil lens in if it is needed.
Gordon
Gordon Couger gcouger-at-couger.com www.couger.com/gcouger Stillwater, OK 405 624-2855 GMT -6:00
At 02:38 PM 02/05/99 +0300, you wrote: } Hello everyone, } } What is the average lifetime of a hair-pin tungsten filament? } We have a JEOL-840A SEM which we operate at 20 KV, and which we run at } asturation (on average) for ~ 7 hours everyday 5 days a week. For the } past several years, filaments used to last 6 weeks or longer, but } recently, the filament is burning out every 5 - 7 days! What could be } the reasons for this, and what are the solutions? } Many thanks. } } Aley El-Shazly } Department of Earth Sciences } College of Science } Sultan Qaboos University } POBox 36, Al-Khod PC 123 } Oman } e-mail: aley-at-squ.edu.om
I get about 50 hours of life from a standard W filament. I can get over 225 hours life from an Energy Beam Sciences SG filament. A good vacuum is very important no matter which filament make you use. I use an ion pump routinely and switch back and forth between W and LaB6.
I'm not sure which type filament your Jeol uses but here are the two types that are available for Jeol:
I did double labelling of whole retinal tissue years ago. The method came from the refs below. It was fairly easy, though it was necessary to be rigorous with controls (as in any double labelling). I didn't persevere as fixation wasn't great and labelling intensity was low. I could probably have improved it, but didn't feel it was worth it. The papers below had (from memory; I no longer have the originals) good results.
Sako H, et al (1986) Simultaneous detection of B-cells and T-cells by a double immunohistochemical technique using immunogold-silver staining and the avidin-biotin-peroxidase complex method. Histochemistry 86:1-4
van den Pol AN (1985) Silver-intensified gold and peroxidase as dual ultrastructural immunolabels for pre- and postsynaptic neurotransmitters. Science 228: 332-5
van den Pol AN (1986) Tyrosine hydroxylase immunoreactive neurons throughout the hypothalamus receive glutamate decarboxylase immunoreactive synapses: a double pre-embedding immunocytochemical study with particulate silver and HRP. J Neurosci 6:877-91
van den Pol AN, Smith AD and Powell JF (1985) GABA axons in synaptic contact with dopamine neurons in the substantia nigra: double immunocytochemistry with biotin-peroxidase and protein A-colloidal gold. Brain Res 348:146-54
Chan J, Aoki C and Pickel VM (1990) Optimization of differential immunogold-silver and peroxidase labelling with maintenance of ultrastructure in brain sections before plastic embedding. J Neurosci Methods 33:113-27
Demonstration of two antigens using a novel combination of immunogld-silver staining and immunoenzymatic labelling. J Histochem. Cytochem. 38:307-13
Krenacs T, Laszik Z and Dobo E. (1989) Application of immunogold-silver staining and immunoenzymatic methods in multiple labelling of human pancreatic Langerhans islet cells. Acta Histochem. 85:79-85
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
Does anyone out there in cyber space have a control board (90014438-611) = for an LKB 2188 NOVA Ultratome. We are quite willing to pay for it as = our ultratome is now off the air. Any suggestions would be gratefully = received.
Terry A Robertson
=20 Dr Terry Robertson (PhD) Senior Research Fellow Department of Pathology University of Western Australia Nedlands 6907
Hi All, I have two boxes of brand new JEOL E-type filaments here. I have no idea what they are doing in the lab: there has never been an instrument that uses these in the vicinity. Anybody have a use for them? Cheers, Malc. Dr MP Roberts Department of Geology Rhodes University Grahamstown 6140 South Africa Tel: +27 46 6038316 Fax: +27 46 6229715 ******************************* "If God had meant birds to fly, he would have given them engines" Anon.
I guess you will have a mass of replies on this one?
It is my experience, working around the world, that filament life almost=
seems more important in many laboratories than the quality of the final image ;-).
What sets the filament life?
1. The magnification level and the kV you wish to use - as you go higher in magnification (} 20,000X) you need to work with a better formed=
probe with as many electrons as possible. Therefore high magnification means on a JSM 840 100 to 120 uA of current, more current means less life= ? =
If you are running to attain true surface images you will find the gun is=
far less efficient at {10kV so the filament life will fall as you drive t= he filament even harder to get the emission current up. Low magnification( {5,000X), when the instrument is not really being used very=
hard at all, the filaments should go on for ever (see later).
2. The way you saturate and align - a wave form is the best method a= s a wide variety of people all come to the same conclusion with this method= . =
However as the filament thins with use, requiring less current, you shoul= d be rechecking your version of saturation and alignment each time you swit= ch the kV on and after each kV change. Then we come on to under saturating!= =
Fine if you run at very low magnifications and do basic x-ray analysis, b= ut do not complain when your images are not so good, poor source production equals poor image production as the final beam spot mimics the source!.
3. The vacuum level - you may fudge your saturation or find excuses for running at a low emission current but the vacuum system cannot be fudged. The life of a filament is directly related to the vacuum level i= n the gun chamber, the better the vacuum the better the filament life! A dirt gun chamber will also "spoil" the gun vacuum so a clean chamber is also important. Remember vacuum gauges tend to be a long way from the gu= n and do not represent its vacuum! JSM840 has the advantage of using an exchange airlock which should give a very good column vacuum.
4. The quality of the filaments - having worked for a number of manufacturers we found from time to time we were supplied from the factor= y with poor quality filaments. They seemed to be made up of wire plus rubbish and they only lasted about 10 hours making a mess of the cathode and gun chamber. We heard about some problems but you will be surprised how few boxes came back. Standard practice should be NEVER use a complet= e box of filaments if they are working well, always save two. When you hav= e a new box of filaments, where the first one gives a poor filament life, u= se one more and work with particular care. If this life too is poor fit one=
of your "good box" filaments and try again. "Good box" filament gives go= od life - problem =3D new box of filaments. "Good box" filament gives poor = life - problem =3D gun vacuum.
5. How you measure it - if you run an instrument that has a meter fitted to the filament on switch you soon learn that even if the instrume= nt is "on" for seven hours a day the actual filament time is far short of that! Many checks have shown that in a seven hour day the filament is unlikely to be on for more than half the time with a single specimen exchange system. With a multi user facility, with operator changes, the filament time is reduced even further. X-ray analysis is the biggest ti= me consumer as people leave the filament on when playing with their spectrum= . =
HT on makes sense filament on costs money!
So to answer the question what is a good filament life? Well it depends
SEM } 50,000X a pointed filament will last about 10-15 hours a V about 20-= 30 hours ~20,000X a V about 30-40 hours {5,000X a V about 40-60 hours EDX a V in excess of 60 hours
TEM it is a different game as we do not drive the gun hard under many applications and to make cross over easy to recognise we run slightly und= er saturated. So TEM depending on application and instrument age 70 hours plus.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
I'm afraid that I didn't see the original posting on this, so I am relying to this reply.
I find it interesting that this thread should run concurrent to a thread regarding the value of service contracts. You often don't kow what you may be missing...
A simple tungsten filament should work for around 80 hours of use. If you are getting less than this, you are either operating the filament at too high a temperature or there are vacuum leaks in your system that permit air at a location that reduces the vacuum in the gun. The major gun seal is opened when replacing filaments and should be checked. A typical SEM vacuum system puts the pumps at one end of the volume being evacuated and the electron gun at the other end. Small leaks in the gun, or in the standpipe that evacuates the gun, can have enhanced effects on the filament life. The standpipe is often the culprit. This is the vacuum connection between the specimen chamber and the electron gun that is provided to enhance the pumpdown of the gun and prevent the gun being pumped down through the column and its apetures.
The standpipe o-ring seals at the specimen chamber and the gun are often a weak point in design. Manufacturers have used a variety of seal designs at both the chamber and gun that often are marginal at best. Another, insiduous source of problems are the seals at the secondary electron detector, EDS detector or other chamber ports. Since they are often near the standpipe opening to the specimen chamber, they to can lead to poor gun vacuum.
If your instrument has a gun translation system, this should also be checked. The gun translation provides for the mechanical adjustment of the position of the electron gun, usually through opposing set screw adjustments at the top of the electron column. Many, if not most, recent model instruments have electronic controls, rather than mechanical controls, that don't present this problem.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, IL 60174 PH 630.513.7093 FAX 630.513.7092 Email: ars-at-sem.com
-----Original Message----- } From: Dr. Gary Gaugler {gary-at-gaugler.com} To: MSA listserver {Microscopy-at-sparc5.microscopy.com}
John - your concerns are unfounded. Modern immersion fluids are essentially non-toxic and they adhere because of extreme surface tension makes them quite sticky and that would not change outside gravity. They are also essentially non-drying and would last in a reduced atmosphere with no appreciable drying. In a clean environment the medium need only be wiped off occasionally. In gravity or with g-forces applied, a hanging drop could move, but that is unlikely to apply in space and during blast-off presumably no oil would be on the objective. If there was concern about the drop of immersion oil being dislodged, I would recommend Cargille's Type NVH with high viscosity of 21,000cST , this type is used for inclined and inverted optics. The drop will never fall off, but it is harder to wipe off. Sure you can do away with oil immersion, but in light microscopy this will lower top resolution attainable.
Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Monday, May 03, 1999 11:15 PM, John Eustace [SMTP:John.Eustace-at-lerc.nasa.gov] wrote: } } Gentlemen, } } I work for a NASA contractor whose job it is to design and } construct } experimental packages for use in space. Currently we are } designing a } package which will make use of a Leica microscope to be } flown aboard the } International Space Station. } } My question regards the use of immersion oil in } conjunction with an } objective. We have purchased an objective designed to be } used as such, and } it's operation is understandable on ground. What I need to } know is if there } is any one who has done any rersearch into the wetting } properties of these } types of oils, such that when we try to deploy them in } space we can wet the } surfaces of interest, i.e. the sample slide, and the } objective? The } necessity for this type of information becomes evident } when you realize } that there will be no gravity to assist in the deployment } of the oil droplet. } } Any assistance in the this matter would be greatly } appreciated. } Thank you in advance } } John } John Eustace } Optical Physicist } Dynacs Engineering } 2001 Aerospace Parkway } Brookpark, Ohio 44142 } Ph (216) 977 - 1244 } Fax (216) 977 - 1269
I'm doing strange observations on a group of tiny mites and I have found that they are able to produce fine drops of idrophobic substance ("saliva" ?) when they are immersed in oil. I have tried some types of oil but "immersion oil for microscopy (ordinary use) nd =3D 1.516 at 23=B0C produced= by Olympus optical Co., LTD" is the only one in which it happens.
My knowledge about the composition of this oil was that it is produced from cedar. So, I tested condensate cedar oil but the mite died soon (after 30 minutes) without any dropplets visible.
My question is:
What is the exact composition of the immersion oil?
Thanks for all help.
dr Enrico de Lillo Istituto di Entomologia agraria - Universit=E0 Bari - Italy tel. +39 080 5443105 fax +39 080 5442876 email: delillo-at-agr.uniba.it http://193.204.185.103/de_lillo.htm
Ann Lehman says: } Couldn't you duck the issue and use high/dry objectives instead? These } offer VERY good optics without the necessity for immersion oil.
Hmm, could you explain how to duck the issue of theoretical impossibility to achieve N.A. greater than 1.0 in the air, with the consequent effects on resolution and meaningful magnification? Also, all other conditions equal, immersion objectives tend to beat dry ones when resolution and magnification are important - so "VERY good" becomes "MUCH better" with oil... High/dry is likely to be 40x to 63x... Oil is likely to be 90x to 100x...
Or have you seen a "very good" high/dry 100x na=1.0 objective?
John, you may need oil-on-oil, i.e. placing the oil-drop on the top condenser lens to oil the specimen slide to the condenser, and oil-drop on the cover-slip to oil the front lens of the objective to the cover-slip. Otherwise your resolution will suffer.
In zero-G your oil droplet will probably just stay a ball so you'd have to "squash" it over the surface to be oiled... This is my rough guess... I'd really like to hear the "actual" solution you eemploy. -- Regards, Uri uri-at-watson.ibm.com -=-=-=-=-=-=- {Disclaimer}
Have solid-state annular BSE detectors for 15kv+ SEM applications attained true TV-rate performance in recent times? If so, could a manufacturer be mentioned, please?
I can't speak for the 'Scope on a Rope, but I saw an interesting all-in-one device from Cole Parmer at PITTCON. It is a digital microscope system which has a 4 MB smart card for collecting images and a 2"liquid crystal screen. It has its own set of lenses (30x comes standard; 1x, 50x, 100x, and 200x also available) so it can act as a traveling microscope for field use but also connects directly to a microscope via a C mount. It also has an adapter kit for interface with a PC or MAC. While not inexpensive, it is a neat package.
Cole-Parmer can b reached at 800-323-4340 or www.coleparmer.com.
I look forward to hearing more about 'Scope on a Rope.
CAVEAT: MME has no financial interest in this product.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 03:39 PM 5/3/99 -0700, Jon Krupp wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} At a recent MSA meeting (within the last 5 years or so) there was a session } on microscopes in the classroom that featured something affectionately } referred to as a 'Scope on a Rope'. } } It was a video camera tethered to a TV with a built in light source and } medium mag. All you did was hold it up to something and a magnified picture } appeared on the TV. } } Of course, now that I need to know more about it, I can't find any } references. Anyone remember it or have an idea of where to start looking? } } As always, thanks a million.
Jon Krupp
Hi, Jon. I believe Carolina Biological sells it. You can get info from Bill & Cindy Henk of Louisiana (check the MSA membership list), who presented it at MSA. It's quite expensive, so the Ken-A-Vision version (Insights, listed on the MICRO page under "microscope suppliers" is one source) is more popular for precollege use. But it must be mounted on a microscope. } Caroline
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
Check out the following web site. Richard Sherman is the patent holder, sells the guns, and can give you reprints of the articles. (Look in JVST circa 1992 for them.) His Email address is co2clean-at-aol.com
A colleague of mine works on primary cortical cultures. When she does immunocytochemistry on them she usually uses indirect (immunofluorescence) method, which works well. When she uses fluorescence labeled avidin-biotin method and she gets extremely high background. I have looked at the controls. The non specific binding is as bright as the specific one. I have never worked with cultures. Could there be endogenous biotin in them? Or would you have any other suggestions on what could be the cause and what can one do to prevent this? Thanking you in advance for your input, Lilith -------------------------------------------------------- Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
Is there a blocking step used in either procedure. When I was doing IHC on TC we used to routinely block, whether it was with a serum or a detergent such as Tween 20. We had success with blocking and felt the background was just a cell culture thing, but were thankful that it was virtually eliminated.
Mary Sr. Lab Tech Bone Marrow Transplant Medical College of Wisconsin Mary_M-at-bmt.mcw.edu
On Tue, 4 May 1999, Barry, Lilith wrote:
} Date: Tue, 4 May 1999 14:48:00 -0400 } From: "Barry, Lilith" {Lilith.Barry-at-nrc.ca} } To: Histonet {histonet-at-Pathology.swmed.edu} , } microscopy {microscopy-at-Sparc5.Microscopy.Com} } Subject: Tissue culture background } } A colleague of mine works on primary cortical cultures. When she does } immunocytochemistry on them she usually uses indirect (immunofluorescence) } method, which works well. When she uses fluorescence labeled } avidin-biotin method and she gets extremely high background. I have looked } at the controls. The non specific binding is as bright as the specific one. } I have never worked with cultures. Could there be endogenous biotin in them? } Or would you have any other suggestions on what could be the cause and what } can one do to prevent this? } Thanking you in advance for your input, } Lilith } -------------------------------------------------------- } Lilith Ohannessian-Barry } National Research Council } Institute of Biological Sciences } CANADA } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca }
I am to prepare samples for TEM from Au 38at% Ni 100um. thick foil.
I will be using a Fischione polisher.
Does anyone have a polishing solution for this alloy?
Thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
I have a paper from Bernie Kestel at Argonne National Lab which describes=
the use of a non-acid electrolyte for thinning Au with the South Bay Technology Model 550 Jet Polisher. The recipe is for his BK-2 solution a= s follows:
The sample polished was annealed, polycrystalline gold and it was thinned=
at -55 degrees C with a potential at 150V at one half the maximum electrolyte flow rate. I believe the Fischione unit only goes up to 120V= , so you may need to adjust the recipe to account for the lower voltage. =
I have been presented with a new challenge and I need to be brought up to speed on a few things.
I need to help some biochemists look at something they call liposomes. Actually they call them SR vesicles and BR vesicles, sensory and bacterial rhodopsin vesicles. This is what I know: The liposomes are in 4M salt, they are supposed to be something like 15 nm in dia. and all they want to know is whether they are intact spheres or broken fragments. Previously they had someone look at them using freeze fracture, but that person is no longer available and we do not have freeze fracture available here.
They suggested negative staining, but they are short on references and advice. I was just going to dive in and do a simple negative stain with UA, but thought I better look for help wherever I can get it.
I am not sure about a couple of things. Does anyone have advice about how to apply the liposomes to the grid, I thought I would just drop some onto a formvar coated grid to start. The 15 nm size bothers me too. Seems small for our TEM and telling one little tiny blob from another little tiny blob is no fun for me.
Does anyone have advice about how the 4M salt will work out. I have visions of looking at salt boulders rather than liposomes. I asked about rinsing out the salt, but they say that will burst the liposomes.
Yes, I do remember a thread about liposomes here recently. Of course, as usual, I did not give it the attention it deserved given my new assignment. Perhaps someone could refresh my memory and clue me in.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
My company is considering to send out samples on a regular basis for EDS analysis with high spatial resolution (in the nm range). I would appreciate to be contacted direcly by email with referrals of analytical labs with FE-TEM or FE-STEM capabilities willing to offer this service.
Adsorption of Poly(2-vinylpyridine) Wormlike Polyelectrolyte Brushes on Mica Studied in-situ by MAC Mode(tm) AFM
U. Schmidt~, S. Prokhorova+, S.S. Sheiko+, M. Möller+, P. Dziezok*, M. Schmidt* ~Molecular Imaging / Roper Scientific, Sollner Str 61, D-81497 Munchen, Germany +Dept. of Organic Chemistry III, Univ. of Ulm, Albert Einstein Allee 11, 89069 Ulm, Germany. * Inst. of Physical Chemistry, Univ. of Mainz, Jakob-Wedler Weg 11, 55128 Mainz, Germany.
Bart Cannon wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi, } } Have solid-state annular BSE detectors for 15kv+ SEM applications } attained true TV-rate performance in recent times? If so, could a } manufacturer be mentioned, please? } } Thanks. } } Bart Cannon
Bart, Either a Robinson Detector or a GW Electronics detector should work at TV rates. Anybody else have any candidates?
Ken Converse owner Quality Images third party SEM service Delta, PA
While on the filament "thread" (oops, a pun slipped in), I am wondering if someone can make recommendations for suppliers of LaB6 filaments for use in a Hitachi H-7100FA (hi res, small probe). Hitachi makes a wonderful assembly but it is getting pricey ($3,500) and we would like to explore other possibilities. The purse keeps getting smaller .....
Thanks, John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
I do agree with several of the respondents that, though filament life can be influenced by a number of factors, the most common cause of an abruptly decreasing life is likely to be poor vacuum. However, I do not recall anyone mentioning one of the simplest ways to check for this condition: by inspecting the tip of the expired filament under a low-power microscope (or use your SEM if you like to indulge in overkill).
Under optimal conditions, a filament's life is determined exclusively by the evaporation of the tungsten. Under optimal conditions (i.e., good vacuum and the filament drive current is not so high that the wire melts), the filament wire will gradually thin as the tungsten evaporates away. But because the thickness of the filament wire is not perfectly uniform and the temperature distribution is also not perfectly uniform, some parts of the filament wire will evaporate faster than others. Eventually, a localized region of the filament wire will have thinned enough so that it will become appreciably hotter than the rest of the filament, this causes the local rate of evaporation to accelerate even more -- the local thin spot has a higher resistance which makes it hotter, which makes it evaporate faster, which makes it thinner, which makes it hotter, etc. -- a regenerative process which quickly results in enough of a local "hot spot" to melt the tungsten and the filament breaks open at this point. The dynamics of filament construction are such that the regions just to either side of the apex of the "V" tip are naturally going to be a little thinner than the tip itself. Thus, when a filament expires "normally" (i.e., solely by evaporation) you will see that the break is on one of these legs -- the wire adjacent to the break will be noticeably thinned to a taper and there will likely be a small "ball" on either side of the break where the molten tungsten solidified. (A pair of large globs of tungsten with negligible thinning indicates that the filament was blown by a high current surge -- just the way a common fuse wire blows.)
Bad vacuum always produces a different kind of filament failure. This is because gas molecules in the gun will be ionized by collisions with the electron beam. The positive ions are accelerated towards the cathode and are focused into the filament tip. This impact sputters the tungsten tip and creates a crater. A filament which has failed because of poor vacuum, when inspected under a microscope, will look like someone took a bite out of the very tip of the "V". There is negligible thinning of the wire along the legs.
These effects are very easy to recognize with a small amount of practice and practice in doing so should really, at least in my opinion, be part of the training for anyone who operates a tungsten filament microscope (similar effects can also be observed for LaB6 filaments). ------ I do want to take a small exception to several of the statements that were made regarding altering the filament life by varying filament height, emission, magnification, kilovoltage, and the like. My objection is not that these statements are not in some sense true, but rather that they confuse cause and effect. Saying that dropping the filament height in the wehnelt will lengthen its life is sort of like stating that putting out-of-balance wheels on your car will result in better gas mileage -- it will -- because you will need to drive slower.
With a properly constructed and operated tungsten filament (i.e., assuming negligible gas sputtering), the rate of evaporation (and thus the life of the filament) are dictated solely by two factors: (1) the temperature of the filament; and (2) the thickness of the wire -- PERIOD. There are no other operational factors.
If you want long filament life, use filaments which have a somewhat thicker wire (many microprobes use this strategy) or simply run the filament at a lower temperature. So why doesn't everyone simply use thick filament wire and/or low temperature? Because these both result in sub-optimal optical performance. In other words, you have a choice: (a) operating your filament for high brightness and best resolution and accepting short filament life; or (b) operating at a lowered temperature for long filament life and accepting reduced source brightness and poorer imaging.
So why do so many microscopists state that they can alter their filament life by changing the height of the filament, changing the emission, altering the kV, etc.? Because these changes can shift the operating point at which "saturation" of the filament is achieved -- thus when the operator subsequently saturates the filament by adjusting the filament drive, he/she is actually changing the filament temperature. Thus, changing one of these parameters may in fact be a practical means of achieving longer filament life, but it is strictly a secondary effect. If you doubt this, I propose a simple experiment: instead of adjusting the filament drive so as to achieve saturation, instead maintain constant filament drive. If you do this, you will find that your filament lifetime is unaffected by any of these other operating conditions (though the quality of your imaging may well deteriorate unacceptably).
There is nothing magic about saturation. It was shown by Haine back in the '30s that the effect which we refer to as "saturation" is solely a function of the way in which the bias resistor circuit regulates the filament's emission. By varying the bias resistor, you can make the filament saturate at any temperature you please. Of course, you are still limited by the fundamental fact that a long-lived filament is a not-so-bright filament (no social commentary intended). But if a long filament life is more important than maximum image quality, then this is a good strategy.
I know these things have been discussed on this listserver before, but they seem to bear repeating in the context of the question which was asked and the discussion which has followed.
Many of my friends/clients have been waiting for me to say this so here goes -
"Just what do people get from an SEM which gives 100 hours filament life?= "
When I visit labs who need to get more from their SEM the problem always = is that they get 100+ hours life (by their means of measurement) and the images die at } 3,000X. The first thing we have to do is give them more current and throw away the filament life. As I have said so many times before "in some labs filament life is more important than image quality!"=
When I am abroad the fuel consumption of my car is amazing, it sits in th= e garage at home and does nothing, should I boast?
Come on guys lets start putting realistic figures on filament life rather=
than hours in a day and lets relate this to the maximum magnification we use and the lowest kV we use. Then and only then will we all have a base=
to work from!
Try this equation - multiply the actual filament on time by the thousand digits of the maximum magnification used and the emission current , then divide by the average kV you use. My typical customer would have a 6.7K value.
I think that would be interesting, anyone with other ideas?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain
I want to thank all of you who responded (on- and off-line) to my question about magnetic samples in the TEM. I got many usefull advises. I will keep all these messages, if anybody is interesting in getting a summary, just let me know!
Dr. Nathalie Bozzolo Laboratoire d'Analyse des Materiaux Centre de Recherche Public - Centre Universitaire 162a, avenue de la Faiencerie L-1511 Luxembourg tel : (352)46 66 44 402 fax : (352)46 66 44 400 e-mail : Nathalie.Bozzolo-at-crpcu.lu _________________________________________________
} From: "drose-at-wlgore.com"-at-Sparc5.Microscopy.Com To: Microscopy-at-Sparc5.Microscopy.Com Date sent: Tue, 4 May 1999 09:43:03 -0400
Greetings. This may be a bit out of the main-stream of our usual concerns but I have a question that many of us may share. In my microscopy lab we have an older model Sorvall Superspeed centrifuge, useful for all sorts of things. I have funds to purchase a new fixed angle rotor but cannot decide on a standard aluminum rotor or one of the new carbon fiber composite rotors. Does anyone have experience with the carbon fiber? No brand names need be involved, but do they have any hidden drawbacks compared to the standard? Anecdotes are welcome. Thank you in advance. Bill P.
I maintain the Tips & Tricks site, biologic archives of this listserver. You are correct there was a thread or two but I don't have them posted to web yet so here are the raw messages. If there is ever anything you need and it is not on the web site below, simply ask and I will search the unposted material. There are roughly 300 discussions not yet posted which exceeds what is posted. I will be addressing this in the next few months as the site gets a facelift. Sorry for the flood, but you asked for it
At 04:25 PM 5/4/1999 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} {} { GO GATORS Scott D. Whittaker 218 Carr Hall EM Technician Gainesville, FL 32610 University Of Florida ph 352-392-1184 ICBR EM Core Lab fax 352-846-0251 sdw-at-biotech.ufl.edu http://www.biotech.ufl.edu/~emcl/ The home of " Tips & Tricks "
Hello dear all! I am looking for any information on the microscope, called Grumhauser and on the person who made it. The spelling I have might be incorrect. Best regards, Dima My email address is la_dima-at-hotmail.com
_______________________________________________________________ Get Free Email and Do More On The Web. Visit http://www.msn.com
BSE at TV bandwidths depends heavily on the performance of preamplifier following the detector. We've marketed a black-box add-on for a little over a decade, through one of the mainstream SEM manufacturers. The preamp offers full TV bandwidth at moderate gain, and 0.6 MHz (fuzzy TV) bandwidth at full gain, to allow real time viewing for positioning the sample.
Still wider band performance is possible, but it's been unclear whether there's a need for it - from here it looked like BSE was being supplanted by environmental detectors.
Jon - Liposomes will be invisible among all the salt crystals. I used to wash such isolates in an ammonium acetate solution. This is a volatile buffer and would sublime after application to the grid. However, you may get sufficiently clean (salt free) preparation by: 1 Apply and blot substrated grid repeatedly to liposome solution. 2 Blot 3 Apply to a series of drops of negative stain (try a couple different ones PTA, UA, ammonium molybdate) and blot in between. Its generally a good idea to use this simple method to obtain good distribution of particles. In this case, you need to eliminate the salt and that requires numerous apply and blot cycles. You could also let the grid float for a while on a drop of stain to allow dilution of the salt. Negative staining solutions do not cause osmotic shock - except when dealing with extreme halophiles.
When you finally see the image, don't be disappointed. Anything prepared by biochemists appears to microscopists as if prepared from a festering cadaver. Remaining salt is easily distinguished: its cubic. But how to judge those liposomes: are they uniform and intact? Its always a matter of degree and never a pretty sight. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Wednesday, May 05, 1999 9:26 AM, Jon Krupp [SMTP:jmkrupp-at-cats.ucsc.edu] wrote: } } I have been presented with a new challenge and I need to } be brought up to } speed on a few things. } } I need to help some biochemists look at something they } call liposomes. } Actually they call them SR vesicles and BR vesicles, } sensory and bacterial } rhodopsin vesicles. This is what I know: The liposomes are } in 4M salt, they } are supposed to be something like 15 nm in dia. and all } they want to know } is whether they are intact spheres or broken fragments. } Previously they had } someone look at them using freeze fracture, but that } person is no longer } available and we do not have freeze fracture available } here. } } They suggested negative staining, but they are short on } references and } advice. I was just going to dive in and do a simple } negative stain with UA, } but thought I better look for help wherever I can get it. } } I am not sure about a couple of things. Does anyone have } advice about how } to apply the liposomes to the grid, I thought I would just } drop some onto a } formvar coated grid to start. The 15 nm size bothers me } too. Seems small } for our TEM and telling one little tiny blob from another } little tiny blob } is no fun for me. } } Does anyone have advice about how the 4M salt will work } out. I have visions } of looking at salt boulders rather than liposomes. I asked } about rinsing } out the salt, but they say that will burst the liposomes. } } Yes, I do remember a thread about liposomes here recently. } Of course, as } usual, I did not give it the attention it deserved given } my new assignment. } Perhaps someone could refresh my memory and clue me in. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu } }
John - Don't know about that model Hitachi. We supply "normal" Lab6 cathodes (Kimball Physics), which I understand are as good as any and they are arguably the most robust. I and no doubt any supplier of LaB6 cathodes would love to sell those to Hitachi (or anybody) for less than 20% of your quoted price.
The difference is so large that I wonder. Do those cathodes have the same base as do Hitachi tungsten filaments? Do you know the size of the microflat atop the LaB6 cone? Normal are 15 square micrometers, I assume that for a high resolution TEM you probably would want that small flat. The larger flats, say 40 sq. micrometers are more suited to microprobes where long term stabillity is more important than is greatest brightness. The larger flats are more expensive. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
On Wednesday, May 05, 1999 12:01 PM, John J. Bozzola [SMTP:bozzola-at-siu.edu] wrote: } } } While on the filament "thread" (oops, a pun slipped in), I } am wondering if } someone can make recommendations for suppliers of LaB6 } filaments for use in } a Hitachi H-7100FA (hi res, small probe). Hitachi makes a } wonderful } assembly but it is getting pricey ($3,500) and we would } like to explore } other possibilities. The purse keeps getting smaller .... } } } Thanks, } John } } ########################################################## } ########## } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ###################################################### #### } ########## } }
Thanks for to those who reponded. Below is information on the CO2 Snow Gun.
--------------------------------- Applied Surface Technologies manufactures CO2 Snow Gun. They can be reached at 908-464-6675, email at co2clean-at-aol.com or on the www at www.co2clean.com.
There are many microscopy examples on the www site by using AFM or SEM. Please look at the site to look for similar applications.
If you need more information, just call or email.
--------------------------------- Check out the following web site. Richard Sherman is the patent holder, sells the guns, and can give you reprints of the articles. (Look in JVST circa 1992 for them.) His Email address is co2clean-at-aol.com
Jon, Freeze Fracture is the best answer considering the salt problem. The other alternative is cryo-TEM, best done with Leo/Zeiss energy filtering imaging. We neagtive stain liposomes just as we would any other sample for conventional TEM. What you see are little bubbles. We have also freeze-dried and shadowed or freeze dried and replicated. You may be able to get around the salt problem if you osmicate first. Just a guess. At 04:25 PM 5/4/1999 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
(a) told me about silver membrane filters, which I had not heard of before (b) reminded me of Gelman, who recommended their PTFE membranes.
As to which I use for which application, it's horses for courses!
And now the question: one has a scanned or digitally acquired micrograph. The background is a gently varying grey, with rather low contrast local features (blobs, bacteria, or whatever). The task is to Fourier transform, then filter out the lowest frequencies with a high-pass filter, leaving one with the features on a uniform grey background. One can then increase the contrast to make the features more prominent for printed reproduction. Is there software (Win 95) that can do this? (preferably inexpensive, though we could make arrangements to use plugins for Adobe Photoshop elsewhere, if necessary).
You can see an example of what I'm trying to do on:
http://www.reading.ac.uk/~spsolley/fourier.htm
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I only worry about filament life if I have to change a filament too frequently. Normal operation would have the filament saturated fully. = If the image quality is unimportant and analysis is the main focus then the current is reduced slightly. Image quality is more important than a =A3= 10 filament which only takes a few minutes to swap ( We keep a spare assembl= y ready ).
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Steve Chapman {PROTRAIN-at-CompuServe.COM} To: American {microscopy-at-sparc5.microscopy.com} is } that they get 100+ hours life (by their means of measurement) and the } images die at } 3,000X. The first thing we have to do is give them more } current and throw away the filament life. As I have said so many times } before "in some labs filament life is more important than image quality!= " } } When I am abroad the fuel consumption of my car is amazing, it sits in t= he } garage at home and does nothing, should I boast? } } Come on guys lets start putting realistic figures on filament life rathe= r } than hours in a day and lets relate this to the maximum magnification we } use and the lowest kV we use. Then and only then will we all have a bas= e } to work from! } } Try this equation - multiply the actual filament on time by the thousand } digits of the maximum magnification used and the emission current , then } divide by the average kV you use. My typical customer would have a 6.7K } value. } } I think that would be interesting, anyone with other ideas? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } }
} ... } } } While on the filament "thread" (oops, a pun slipped in), I am } wondering if } someone can make recommendations for suppliers of LaB6 } filaments for use in } a Hitachi H-7100FA (hi res, small probe). Hitachi makes a wonderful } assembly but it is getting pricey ...
I don't know about your SEM but we've had excellent performance and lifetimes using FEI cathodes, LaB6 and CeB6 ... see:
http://www.feibeamtech.com/lab6/lab6page.htm
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Is she using Thermanox coverslips to grow the cells on? If so, they exhibit bright autofluorescence. Don't know what the coating is. Ronnie Houston Cytochemistry & Molecular Pathology Texas Scottish Rite Hospital for Children Dallas, TX
Barry, Lilith wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } A colleague of mine works on primary cortical cultures. When she does } immunocytochemistry on them she usually uses indirect (immunofluorescence) } method, which works well. When she uses fluorescence labeled } avidin-biotin method and she gets extremely high background. I have looked } at the controls. The non specific binding is as bright as the specific one. } I have never worked with cultures. Could there be endogenous biotin in them? } Or would you have any other suggestions on what could be the cause and what } can one do to prevent this? } Thanking you in advance for your input, } Lilith } -------------------------------------------------------- } Lilith Ohannessian-Barry } National Research Council } Institute of Biological Sciences } CANADA } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca
I looked in some FIM books for polishing solutions. I found one for gold that might work on your alloy.
for gold: equal volumes of conc. HNO3 and conc HCl, 1-10Vac
Note 10%(5-10Vdc)HCl will work for Ni (I also found 40% solution at 1-2Vdc) so this might work for your Au-Ni alloy.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Fred Pearson To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hello Microscopists:
I am to prepare samples for TEM from Au 38at% Ni 100um. thick foil.
I will be using a Fischione polisher.
Does anyone have a polishing solution for this alloy?
Thanks in advance
Fred Pearson
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
There is an old analog trick of unsharp masking which might give you a similar result. You make an out-of-focus contact print of your original negative, then sandwich the two together in your enlarger to make your final print. This effectively subtracts the low frequency background from the original image and adjusts the contrast of fine details in high-contrast images. My reference was Kodak Techbits 1990, issue 1 (pub # P-#-90-1).
You can mimic this analog technique by using a very large blurring kernal on the original and subtracting the blurred image from the original image. If necessary, you can multiply the blurred image by a constant to adjust the final intensity. You can also define a custom kernel to get something large enough to do what you want. This should help even out the background in your image.
Henk
At 03:17 PM 5/5/99 +0100, you wrote: } } } And now the question: one has a scanned or digitally acquired micrograph. } The background is a gently varying grey, with rather low contrast local } features (blobs, bacteria, or whatever). The task is to Fourier } transform, then filter out the lowest frequencies with a high-pass filter, } leaving one with the features on a uniform grey background. One can then } increase the contrast to make the features more prominent for printed } reproduction. Is there software (Win 95) that can do this? (preferably } inexpensive, though we could make arrangements to use plugins for Adobe } Photoshop elsewhere, if necessary). } } You can see an example of what I'm trying to do on: } } http://www.reading.ac.uk/~spsolley/fourier.htm } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ }
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu "Progress does not consist in replacing a wrong theory with a right one. It consists of replacing a wrong theory with one that is more subtly wrong."
Looking for an old JEOL TEM 2000FX. Please let me know if someone has a clue of a place who want to give away their old TEM for free to a University (we will pay the frieght). A JEOL 2000FX TEM even if it is not in working order would be o.k. as we just need it for parts as a back-up for other TEM (same model) that we have.
Please reply directly on my e-mail: zur-at-mmae.engr.ucf.edu
Thank you.
Zia ur Rahman Electron Microscope Engineer University of Central Florida, Orlando, Florida
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Reply to: RE: Tissue culture background Dear Barry, This sounds like a case of the wrong blocking agent. From what you = describe, my guess is your colleague treats the cells with dilute serum (= BSA or FCS) prior to addition of the fluorescent marker. It also looks as = if she does not dilute the fluorescent avidin in blocking agent either. The reason for the strange result is that serum contains biotin-like = molecules. When applied to the cells it will stick all over and the = fluorescent avidin will bind to it giving high background). Try the = labeling protocol without blocking agents and see if there is a difference.= Alternative strategies include substituting a non-serum blocker (such as = cold-water fish skin gelatin) for the serum, or even using antibodies to = biotin as a replacement for the avidin. Biotin-like molecules are also = present in mitochondria. If the cells have an unusually large number of = mitochondria in them, then this too could give the "high background" = described. Regards, Paul Webster
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Barry, Lilith wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
id {2LL9JGKA} ; Wed, 5 May 1999 13:39:29 -0500 Message-ID: {D8F9EBE8536ED111920F00005A422A315A474A-at-commserver.srrc.usda.gov}
Fred makes a good point about examining the tip end of an "expired" tungsten filament. Besides indicating possible vacuum problems, the size of the metal ball at the tip of the filament (usually on the shorter tip) can also indicate user error during filament saturation.
} ---------- } From: Fred Schamber (personal)[SMTP:fhscham-at-sgi.net] } Sent: Tuesday, May 04, 1999 10:27 PM } To: Listserver, Microscopy } Subject: Re: filament life } } } I do agree with several of the respondents that, though filament life } can be influenced by a number of factors, the most } common cause of an abruptly decreasing life is likely to be poor } vacuum. However, I do not recall anyone mentioning one } of the simplest ways to check for this condition: by inspecting the tip } of the expired filament under a low-power } microscope (or use your SEM if you like to indulge in overkill). } } The dynamics of filament construction are such that the } regions just to either side of the apex of the "V" tip } are naturally going to be a little thinner than the tip itself. Thus, } when a filament expires "normally" (i.e., solely by } evaporation) you will see that the break is on one of these legs -- the } wire adjacent to the break will be noticeably thinned } to a taper and there will likely be a small "ball" on either side of the } break where the molten tungsten solidified. (A pair of } large globs of tungsten with negligible thinning indicates that the } filament was blown by a high current surge -- just the way } a common fuse wire blows.) } This "high current surge" can often be found with SEM's which have multiple users. Especially with older and less expansive instruments, the tip failure shape will indicate oversaturation by less experienced users. This is less of a problem with those instruments with "lockout" features where the primary user can set the filament saturation limits on the SEM.
} Bad vacuum always produces a different kind of filament failure. This } is because gas molecules in the gun will be ionized } by collisions with the electron beam. The positive ions are accelerated } towards the cathode and are focused into the } filament tip. This impact sputters the tungsten tip and creates a } crater. A filament which has failed because of poor } vacuum, when inspected under a microscope, will look like someone took a } bite out of the very tip of the "V". There is } negligible thinning of the wire along the legs. } } These effects are very easy to recognize with a small amount of practice } and practice in doing so should really, at least in } my opinion, be part of the training for anyone who operates a tungsten } filament microscope (similar effects can also be } observed for LaB6 filaments). } If the instrument does not have multiple users or new users, I would check for vacuum leaks as a possible cause of repeated filament failure.
Bruce F. Ingber Biologist- Electron Microscopy USDA-ARS, SRRC 1100 Robert E. Lee Blvd. New Orleans, LA 70124-4305
Hello!=A0=A0 We are looking for some help in choosing and/or finding an objective for our Nikon Diaphot scope.=A0 We use the scope (setup for epifluorescence microscopy) to measure the response of neurons to mechanical deformation as a model for traumatic brain injury.=A0 We currently use either Fura-2 (calcium) or Di-8-ANNEPS (membrane potential) to measure the cellular response while the mechanical injury is applied to the cells by a device that is attached to the microscope.=A0 The trouble we're having is that the cells are attached to an elastic membrane and they need to be in saline during the experiment.=A0 Since the microscope is inverted (it's a long story), we think the best way to image the cells is with a water immersion objective.=A0 However, our current objective is a 40X oil immersion and we are using it in liquid (not the ideal situation).=A0 We have funds for a new objective and we'd like your input as to what would be the best objective given our current hardware (old Nikon with 160 mm tube length) and device (cells must be immersed in saline and there is no coverslip) limitations. Another requirement is a long working distance since we need to be able to change the medium between the objective and the cells during an experiment.
What we think we need is a high magnification (40X or greater) fluor water immersion objective with a long working distance on the order of millimeters. We know that newer objectives are made to meet these needs but they do not fit our old scope. Do you have any ideas or recommendations for objectives?
Thanks in advance,
Donna M. Geddes GT/Emory Department of Bioengineering Georgia Institute of Technology Atlanta Georgia, 30332
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I agree Steve.
I only worry about filament life if I have to change a filament too frequently. Normal operation would have the filament saturated fully. = If the image quality is unimportant and analysis is the main focus then the current is reduced slightly. Image quality is more important than a =A3= 10 filament which only takes a few minutes to swap ( We keep a spare assembl= y ready ).
Colin
Colin Reid, Electron Microscope Unit, Trinity College Dublin, Dublin 2, Ireland. Tel: 353-1-6081820 Fax: 353-1-6770438 email: creid-at-tcd.ie -----Original Message----- } From: Steve Chapman {PROTRAIN-at-CompuServe.COM} To: American {microscopy-at-sparc5.microscopy.com} is } that they get 100+ hours life (by their means of measurement) and the } images die at } 3,000X. The first thing we have to do is give them more } current and throw away the filament life. As I have said so many times } before "in some labs filament life is more important than image quality!= " } } When I am abroad the fuel consumption of my car is amazing, it sits in t= he } garage at home and does nothing, should I boast? } } Come on guys lets start putting realistic figures on filament life rathe= r } than hours in a day and lets relate this to the maximum magnification we } use and the lowest kV we use. Then and only then will we all have a bas= e } to work from! } } Try this equation - multiply the actual filament on time by the thousand } digits of the maximum magnification used and the emission current , then } divide by the average kV you use. My typical customer would have a 6.7K } value. } } I think that would be interesting, anyone with other ideas? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } }
Looking for an old JEOL TEM 2000FX. Please let me know if someone has a clue of a place who want to give away their old TEM for free to a University (we will pay the frieght). A JEOL 2000FX TEM even if it is not in working order would be o.k. as we just need it for parts as a back-up for other TEM (same model) that we have.
Please reply directly on my e-mail: zur-at-mmae.engr.ucf.edu
Thank you.
Zia ur Rahman Electron Microscope Engineer University of Central Florida, Orlando, Florida
} We have a Sony DCR-TRV9 digital video camera recording. We would like to } be able to take the video stream, send it to a pc based computer, edit the } file, and grab individual frames and save them as separate files. Has } anyone out there got any suggestions? We'll also need a way to get the } video to the computer. We've tried frame grabbers but because there are } so many frames per seconds, it locks it up. The video is stored on an } Mini DV Digital Video Cassette. } Thanks in advance for any ideas. } } Robin Griffin } UAB } }
Here is the recipe for better performance, a 15 year old SEM is just a youngster!.
1. Use the wave form and the main peak (the one when heating the filament does not increase the signal level) do not undersaturate and ali= gn as accurately as you can
2. You do not say which SEM you have but if it has a bias control adjust the emission current to nearer 100uA. If you do not have a bias control half the distance between your filament and the front face of the=
cathode aperture. If the current is then 70 to 100 at saturation that is=
much better if not shorten the distance again.
3. Work at less than 15mm working distance.
4. Do not worry about how noisy the screen image is, concentrate on getting the best photograph. To do this reduce the spot size (screen goe= s dimmer) until the image when focussed and stigmated looks good on a mediu= m slow scan rate. With a noisy image focus and stigmate for maximum contrast, never stigmate on a directional image.
All this said simply increasing the emission current and shortening the W= D should be more than enough. If you let me know exactly which instrument you use I would be pleased to give specific instructions for that instrument.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
We also have a Hitachi 7100FA, running with a LaB6 . We bought it with two Hitachi cathodes. When these were used we put in a Denka that we had sitting in a drawer - just the straight (M3?) hunk of crystal on a tungsten loop design - and it has worked very well indeed. In spite of being a completely different shape to the Hitachi crystal - short and fat rather than long and thin. We dont use the machine much for really high res work, so I couldnt say if the ultimate performance is different, but we routinely adjust the alignment using a TV camera and screen giving an 8 million X mag, and havent noticed a difference, and the stability has been very good. We always use the auto filament run up and are very conservative - 10 min in the morning, 5 min subsequently, the filament goes off at every specimen and camera change. The Denka performance surprised us, because on an SEM we have had consisitently much better experiences with the Kimball Physics cathode.. .. We cleaned and readjusted the height at about 600 hours, which is similar to what we did with the Hitachis. I dont know if we just dropped lucky with this Denka or what . Is the slow run-up what makes the difference? - so I'm still uncertain what to get when it gives up the ghost! I'd also appreciate hearing what other people's experience has been with LaB6s on the 7100.
regards Sally Stowe
Dr Sally Stowe Facility Coordinator Australian National University EM Unit Research School of Biological Sciences Box 475, ACT 2601, Canberra, Australia FAX 06 (0)2 6279 8525
} } } John J. Bozzola {bozzola-at-siu.edu} 5/05/99 12:00 } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
While on the filament "thread" (oops, a pun slipped in), I am wondering if someone can make recommendations for suppliers of LaB6 filaments for use in a Hitachi H-7100FA (hi res, small probe). Hitachi makes a wonderful assembly but it is getting pricey ($3,500) and we would like to explore other possibilities. The purse keeps getting smaller .....
Thanks, John
#################################################################### John J. Bozzola, Ph.D., Director Center for Electron Microscopy 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html ####################################################################
John J. Bozzola wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } While on the filament "thread" (oops, a pun slipped in), I am wondering if } someone can make recommendations for suppliers of LaB6 filaments for use in } a Hitachi H-7100FA (hi res, small probe). Hitachi makes a wonderful } assembly but it is getting pricey ($3,500) and we would like to explore } other possibilities. The purse keeps getting smaller ..... } } Thanks, } John } } #################################################################### } John J. Bozzola, Ph.D., Director } Center for Electron Microscopy } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Fax: 618-453-2665 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/departments/shops/cem.html } ####################################################################
John, I have a couple of customers who have had good luck with the Kimbal Physics LaB6 tips. They run in the vicinity of $500, last I knew, and they last a long time. They also are not nearly so sensitive to thermal stress as the competing mounting systems, so failure tends to be due to evaporation of the Lab6, not mount failure.
Barry Scientific in Massachusetts may be a little closer to deal with than Jim (sorry Jim!). Kimball no longer retails, but they are in New Hampshire.
Ken Converse owner Quality Images third party SEM service Delta, PA
My company deals in Refurbished Lab Equipment, however we do not do a large business in EM and I admit that I am not well versed in this field. I do from time to time come across used EMs of different brands & styles that I broker.
I do have a few related items in stock, i.e.: a Leica (Reichert) Super Nova Ultra Microtome.
If anyone is interested, please contact me for details.
Ford M. Royer Analytical Instruments, Ltd. 9921 13th Ave. N. Minneapolis, MN 55441 phone: (800) 565-1895, ext. 17 FAX: (612) 929-1895 email: froyer-at-bitstream.net Web Site: www.aibltd.com
Shalvoy, Richard B CHES wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I'm looking for contact information for vendors of used equipment (ie my old } SEM). } } I'm sure they're out there but being pretty new to this area I'm not sure } where to find them. } } Richard Shalvoy } Arch Chemicals (formerly Olin Corporation) } Cheshire, CT
Are there any non volatile fixatives for animal and vegetable material for LM? I am an amateur with very serious asthma. Formaldehyde and formic acid both cause very serious reactions. I don't think it is formaldehyde but some of the less soluble stuff that is formed causing the problem. All my research shows that formaldehyde is too soluble to make it to the lungs. It reacts with the sinuses and throat before it gets to the lungs.
I can't really visualize something that isn't pretty volatile and reactive that would fix tissue. I see references to HgCl, picric acid, acetic acid. potassium permanganate and silver nitrate being used in fixing solutions. I am comfortable handling any of these and none of them cause me a big problem. But I only see them used in conjunction with an aldehyde. Can any one give me some alternatives to the ayldhydes?
While I had several hours of microbiology as an undergraduate the microscope was just one of the tools. With digital image capture and unexpected retirement it opens a whole new world to LM.
I find this list to be an outstanding resource. Nestor and the group do an outstanding job keeping on topic without being heavy handed. The group also seems to be spending a lot less of their employer's time on the net than in most groups. There will be no post during working hours and a flock of messages an evening. I guess that it being an industry mail list I would think twice before posting in working hours knowing that my next resume would probably go to some one on the list.
Thanks Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
A discussion I was engaged in today relative to my last-night's posting suggests to me that there is a bit more to be said regarding the role of emission relative to filament life. There seems to be a common assumption that emission current plays a role in filament life. This is not true.
The typical heating power applied to a tungsten filament in an electron microscope is on the order of 6 watts (product of heater current times heater voltage). Since the filament is operating in a vacuum, this power can be dissipated via three routes: (1) heat conducted through the posts of the filament; (2) photons (light) radiated by the incandescent wire; and (3) energy carried off by emitted electrons (emission current). How large is the latter effect? A WRONG way to estimate this is to take the product of the emission current and the beam voltage. For example, when operating at 30 keV with 100 microamps of emission (a higher emission than most modern scopes use), you would come up with a dissipated power of 3 watts, which is appreciable. This is in fact the power which the gun assembly as a whole imparts to the beam, but it is irrelevant to the question of power dissipation by the filament. This is because that 30 keV of kinetic energy is imparted AFTER the electron leaves the filament.
In fact, an electron is "boiled off" the heated filament with a very low energy. There is actually a distribution of energies, depending on the temperature, but the average is about 1 eV. If one uses 1 eV as the electron energy, then at 100 microamps of emission the dissipated power being carried off through the emitted electons is only 100 microwatts. This is obviously insignificant relative to the five watts of heating power being applied to the filament. Thus, we can conclude that emission plays no role in the filament heat balance and thus does not affect filament life in any practical sense. Furthermore, since the energy at which the electrons leave the filament is independent of accelerating voltage, we also see clearly that the beam voltage plays no direct role in filament life.
So where does the heat go? It has been some years since I did this calculation, but I remember the conclusion that I obtained for one geometry -- heat conducted through the posts is about half of the heat radiated as light. This will, of course, depend considerably on the geometry specific to an instrument, and since I ignored a lot of the complexities of reflected light, etc., my computation was only an approximation, but I think it is safe to say that the radiated and conducted heat components are comparable in magnitude.
I think the reason that so many people believe that emission current is related to filament life is because if they "undersaturate" the filament they observe both a lower emission current and a longer filament life. But both of these are consequences of the fact that they have actually reduced the filament temperature, which is the controlling factor on filament evaporation rate.
We have a well-used but perfectly functional Hitachi S-450 SEM available to anyone who may have a use for it as spares or a fully functional instrument. Cost highly negotiable, transport your problem.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0)331 260 5155 Fax +27 (0)331 260 5776 website:http:www.nu.ac.za (departments} units) Email:bruton-at-emu.unp.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
Synoptics were selling something called `Massram' literally that - a mass of RAM (memory) to allow continuous video to be captured to memory without the problems of access time to any disc storage device. The problem is that storing continuous video really eats up the memory (250Kb? per frame x 25 or 30 frames/sec x time of video sequence). It will be too expensive to buy for a one off but there are some out there somewhere. From memory Bath University in the UK had a system and offered a service but I don't know the cost.
Try Synoptics in the UK for contacts +44 (0) 1223 727100 or fax +44 (0) 1223 727101.
Good luck, Ron
On Wed, 5 May 1999 15:07:33 -0500 "rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com wrote:
} } We have a Sony DCR-TRV9 digital video camera recording. We would like to } } be able to take the video stream, send it to a pc based computer, edit the } } file, and grab individual frames and save them as separate files. Has } } anyone out there got any suggestions? We'll also need a way to get the } } video to the computer. We've tried frame grabbers but because there are } } so many frames per seconds, it locks it up. The video is stored on an } } Mini DV Digital Video Cassette. } } Thanks in advance for any ideas. } } } } Robin Griffin } } UAB } } } } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Does anybody out there know of a spare HT cable for a Siemens Elmiskop 102 TEM? I can't afford to buy a custom built new one, so my only real alternative is to try and track down a second hand one. Can anybody help?
I've used Denka, FEI and Kimball LaB6 emitters in my Jeol 2000. The Denka gave the best image quality but required to be run up very carefully. Out of 4 only one died of old age; the rest were destroyed by careless users, in spite of death threats from me. The FEI and Kimball emitters do not give quite such a good image, but they are really quite robust. I think the Kimball is slightly tougher than the FEI. I've caught people treating them worse than I would treat a tungsten filament, without any ill effects.
I have no financial interst in any of these suppliers.
Regards, Eric
---------------------- Dr Eric E. Lachowski University of Aberdeen Department of Chemistry Meston Walk Old Aberdeen AB24 3UE Scotland +44 1224 272934 e.lachowski-at-abdn.ac.uk
I am looking for informations about nitrogen quantitative analyses by WDX system and specially in biological samples. I am interested also to know the sensitivities (minimum detection limits) and what is the result with biological sample (matrix effect such absorption with about 40% of c and 15% of O). Many thanks in advances.
Didier
-------------------------------------------- Didier Le Thiec I.N.R.A. Centre de Recherches Forestieres Unite d'Ecophysiologie Forestiere Laboratoire de Pollution Atmospherique 54280 Champenoux - France
Thank you for responding to my question on tissue culture background. I don't think the background is because the fixation or the immuno method because I routinely use the same method on brain sections and do not get the same background. It isn't the coverslip autofluorescing either because one can see beautiful labeled structures in the cells except on the controls. So it has to be something specific about cortical cultures and the strept or neutravidin-biotin method. Do you have any other thoughts? Lilith ------------------------------------------------------------------------ Lilith Ohannessian-Barry National Research Council Institute of Biological Sciences CANADA Tel;613-993-6460 Fax;613-941-4475 e-mail; lilith.barry-at-nrc.ca
Ron: As I recall, Texas Memory offers something similar. See http://www.texmemsys.com/ -Mike
Ron Doole wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hi Robin, } } Synoptics were selling something called `Massram' } literally that - a mass of RAM (memory) to allow continuous } video to be captured to memory without the problems of } access time to any disc storage device. The problem is that } storing continuous video really eats up the memory (250Kb? } per frame x 25 or 30 frames/sec x time of video sequence). } It will be too expensive to buy for a one off but there are } some out there somewhere. From memory Bath University in } the UK had a system and offered a service but I don't know } the cost. } } Try Synoptics in the UK for contacts +44 (0) 1223 727100 or } fax +44 (0) 1223 727101. } } Good luck, } Ron } } On Wed, 5 May 1999 15:07:33 -0500 } "rgriffin-at-eng.uab.edu"-at-Sparc5.Microscopy.Com wrote: } } } } We have a Sony DCR-TRV9 digital video camera recording. We would like to } } } be able to take the video stream, send it to a pc based computer, edit the } } } file, and grab individual frames and save them as separate files. Has } } } anyone out there got any suggestions? We'll also need a way to get the } } } video to the computer. We've tried frame grabbers but because there are } } } so many frames per seconds, it locks it up. The video is stored on an } } } Mini DV Digital Video Cassette. } } } Thanks in advance for any ideas. } } } } } } Robin Griffin } } } UAB } } } } } } } } } } ---------------------- } Mr. R.C. Doole } Department of Materials, } University of Oxford. } Parks Road, Oxford. OX1 3PH. UK. } Phone +44 (0) 1865 273701 } Fax +44 (0) 1865 283333 } ron.doole-at-materials.ox.ac.uk
Dear listers, Can anyone help me? I am trying to put a grant application together at very very short notice, and I need to find out the cost of a GOOD TEM. Not one that is the BMW of TEMs, more like an Alfa Romeo - if you know what I mean, and preferably one that has some sort of computerised image assistance with it. Thanks for your help in advance. Please email me as soon as humanly possible at
alex.black-at-nuigalway.ie
Thanks again.
Alex
__________________________________ Alexander Black Department of Anatomy National University of Ireland, Galway Republic of Ireland
via smtpd (for sparc5.microscopy.com [206.69.208.10]) with SMTP; 6 May 1999 20:19:17 UT Received: FROM toto.research.aa.wl.com BY akita.research.aa.wl.com ; Thu May 06 16:12:42 1999 Received: by toto.research.aa.wl.com with Internet Mail Service (5.5.2232.9) id {JNYX3544} ; Thu, 6 May 1999 16:12:41 -0400 Message-ID: {602B9620D104D2118E0A00805FBBC43602826DE0-at-redfox.research.aa.wl.com} {MICROSCOPY-at-Sparc5.Microscopy.Com}
I recommend you contact (if they don't contact you from monitoring this List) a few of the major EM suppliers, especially those from your area. An internet search for Philips, Hitachi, Zeiss, Jeol or any other brand (sorry to those I left out) should get you in contact with a willing sales representative who can work with your urgent time schedule. It will not be easy, since the features are many, and very specific to your lab research focus. Your general query to this list will only get you personal biases, and will likely mislead you more than help you.
Better yet, these companies can fax or express-mail you an official quote to include in your proposal.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Gregg.Sobocinski-at-wl.com
-----Original Message----- } From: ALEX BLACK [mailto:ALEXANDER.BLACK-at-NUIGALWAY.IE] Sent: Thursday, May 06, 1999 2:22 PM To: MICROSCOPY-at-Sparc5.Microscopy.Com
Dear listers, Can anyone help me? I am trying to put a grant application together at very very short notice, and I need to find out the cost of a GOOD TEM. Not one that is the BMW of TEMs, more like an Alfa Romeo - if you know what I mean, and preferably one that has some sort of computerised image assistance with it.
Thanks for your help in advance. Please email me as soon as humanly possible at
alex.black-at-nuigalway.ie
Thanks again.
Alex
__________________________________ Alexander Black Department of Anatomy National University of Ireland, Galway Republic of Ireland
I was a happy user of a plain and standard SEM Zeiss DSM 940, which gave = me nice shots. I work with systematics of sphecid wasps and used to photogra= ph uncoated specimens. I got my best pictures using BSE detection, although = our microscope has no detector for backscatering electrons. Now we have a bra= nd new machine, a LEO 440, with almost all acessories, including a BSE detector. However, I cannot acheive the same results that I got with the Zeiss SEM. As the LEO 440 is wholly automatized, its software do not allo= w me to make the same adjustments that I used in the Zeiss machine. I can u= se only SE detection, that gives me pictures of bugs with a methalic aspect, which is pretty but not usefull. So, let post you two questions. Why a go= t nice pictures in a SEM using a dection method without the proper detector= ? How I can get similar results in a fully coputer controled machine?
S=E9rvio T=FAlio Pires Amarante
serviopa-at-usp.br
Museu de Zoologia da Universidade de S=E3o Paulo Caixa Postal 42694-970 04299-970 S=E3o Paulo BRASIL
Steve Chapman responded to my recent posting with the suggestion that I am missing the point of filament "saturation" and included some further thoughts regarding his theory of how this relates to filament lifetime (quote appended). He concludes with an observation which suggests that "getting it right" in terms of understanding the mechanism is important to operting a SEM effectively. Consistent with that sentiment, I am going to launch into a fairly lengthy discussion of the phenomenon that we know as "saturation", since it is my experience that it is widely misunderstood. I'm also going to touch on some related subjects like the role of "brightness". But please be warned, this is NOT a brief posting. OK?
} I read with interest Fred's note but fear he has missed one important } feature of saturation? } } Saturation occurs when the aperture in the front of the cathode is filled } with electrons, generate more and they are unable to pass through therefore } no change in emission current. } } What affects this aperture? The physical size of the aperture and the bias } field. Apply more bias and the effective aperture size is reduced, } therefore the number of electrons needed to fill the aperture is reduced, } therefore the amount of heat required to produce the required number of } electrons for saturation is also reduced. Hence apply more bias you need } less heat you get a longer filament life as heat (oxidation and } evaporation) is the determining factor. } Steve's analysis is based on the perception that "saturation" is a fundamental electron-optical effect which limits the emission directly. For an electron microscope operated in the normal way, this is simply not the case.
The "kernel of truth" in Steve's argument is that there is indeed an effect of the kind he describes in what are referred to as "high perveance" guns. The effect is due to "space charge" effects encountered when there is a high density of electrons in front of the cathode. Since electrons repell each other, a sufficiently high density of electrons will effectively screen the cathode from the accelerating field, and indeed there will be the kind of effect he describes where you simply cannot add more electrons to the beam.
Unfortunately for his argument, that is not the regime in which electron microscopes operate by design. A microscope designer tries hard to stay away from this condition because the screening action defocuses the beam and increases the energy spread (the Boersch effect ) long before it appreciably limits the emission current. This effect becomes more important at low accelerating voltages since the electrons, being accelerated more slowly, can hang around in front of the filament tip longer. This is why you want to move the anode closer to the filament under low voltage imaging -- you want to increase the field gradient and get those electrons out of there as rapidly as possible. So it is in fact possible to operate an electron microscope under high-perveance conditions where there is a true space-charge "saturation" effect (set the filament way back, the anode at its furthest spacing, and crank the beam voltage way down). But this seriously degrades the imaging and, given any choice in the matter, you want to stay as far away from this operating condition as you can; this kind of aberrant situation is definitely not what people normally mean when they talk about a "saturated filament". So I will stand by my original assertion that the effect that we call "saturation" is simply a consequence of the common bias-resistor circuit, nothing more.
In the early days of electron microscopy, it was generally assumed that the "saturation" condition was something like what Steve describes -- you were "saturating" the phase space available for electrons. Haine soundly disproved this in a 1952 paper (in an earlier posting I erroneously referred to this as being published in 1938) [Haine & Einstein, Characteristics of the hot cathode electron microscope gun, British Journal of Applied Physics, Volume 3, p40, 1952]. On page 45 of this paper he states: "There has been some confusion in discussions on electron guns as to the effect of self-biasing (automatic biasing), the bias being obtained from the voltage drop across a resistance connected in series with the h.v. supply. With this type of biasing, an electron gun exhibits the property, frequently termed 'saturation,' whereby the current rises with temperature to a limiting value beyond which increase in temperature produces little increase in beam current. This has been attributed to the gun running into the space charge limited condition. That this is not so is readily seen when this type of bias is replaced by a variable independent bias; no such effect then takes place." He then goes on to prove his point with graphs. I personally have had extensive experience in working with an electron microscope gun equipped with a separate bias supply and can verify what he states -- there is no "saturation" effect without the bias resistor..
Let's look at the situation in detail (darn, I wish I had a blackboard!). Any electron optics reference will show you a figure of a self biased triode gun. The negative high voltage is connected directly to the wehnelt (grid) and thence to the filament (cathode) via the bias resistor. All current flowing from the filament must pass through the bias resistor, resulting in a voltage drop which makes the cathode more positive than the wehnelt. Thus, interposed between the cathode and the anode is the wehnelt which is at a somewhat more negative potential than the cathode, thus inhibiting the emission of the cathode's electrons. This creates a self regulating situation: any increase in emission current causes the bias to increase, thus choking off emission; a decrease in emission causes the bias to decrease, thus permitting greater emission. The effect is that a constant emission current is maintained, despite minor variations in filament temperature, cathode-tip geometry, and the like. This is a marvelous circuit -- both simple and highly effective with a near-instantaneous response. However, it is not unique to electron microscopy. Anyone old enough to have worked with vacuum tubes has seen it used for a constant current source and any idea about fancy optical effects is clearly not applicable there. To drive the point home: the effect which microscopists call "saturation" is nothing more than reaching the operating point in the emission current stabilization circuit established by the bias resistor -- there is no profound electron optical effect involved. Certainly it is not a matter of the electrons having "filled" a virtual "aperture" established by the bias field.
This may have some readers perplexed at this point. If the "knee" in the saturation curve is caused only by reaching the operating point in a simple emission stabilization circuit, why is this the best point to operate the gun? The most basic answer is: "because you would have to be an idiot to design a gun any other way"! Assuming you know the best conditions for imaging, why would you stabilize your emission current anywhere else?
Let's consider the way the wehnelt does its job of restricting cathode emission. The only reason that electrons can get past the negatively biased wehnelt at all is because the orifice in the wehnelt allows some of the anode's positive field to penetrate to the tip of the filament (the "zero equipotential"). When properly adjusted, there is a small circular region at the tip where electrons can escape to the anode -- everywhere else on the filament, the low energy electrons which are boiled off of it are repelled back into the filament. As the relative negative bias on the wehnelt increases, the anode field doesn't penetrate as far and the size of the emitting region on the filament tip shrinks so that fewer electrons can escape. As the bias decreases, the anode field penetrates further and the size of the emitting region increases, resulting in more emission. The objective of gun biasing is thus to select an emission region right at the tip of the filament. This is common knowledge covered in any optics text.
If you are controlling the bias with an independent voltage supply, you can completely cut off emission by applying a very high bias, or you can create a very high emission (up to the current limit of your supply) by setting the bias at a very low value. What you actually want to do, of course, is set the bias so that just the tip of the filament is able to emit. In this "independent bias" configuration, changing the filament temperature has no effect on the bias, and thus no effect on the position of the emitting region. Increasing the filament temperature causes the emission to increase without limit (and there is no "saturation" rolloff). In this kind of arrangement, it is really easy to understand the relationship of filament life and emission since both are direct consequences of filament temperature.
When you introduce the "self-biasing" resistor circuit, it gets a little more confusing since the effects are coupled. For one thing, you can't set the gun into a "cutoff" condition, because that would require an infinitely large bias resistance. For another thing, the relationship between filament temperature and emission current is different above and below the "saturation" knee. But the fact remains that you do establish a particular bias voltage value via this circuit and for the same bias value, you will get exactly the same emission as with "independent biasing". However, now in order to change the bias you either need to change the bias resistor (R) or to change the filament heating current so as to change the emission current (I) so as to effect the desired I*R drop (bias). But, if you have any sense at all, you select the bias resistor value such that the "saturation" knee occurs at a filament temperature which gives both a nice compact emission pattern AND an acceptable filament lfe. But this is a product of good design practice, not any kind of optical effect. Fortunately, this target is rather easy to hit.
As Steve has pointed out, the effect on the emission pattern of changing the bias can be readily observed on a TEM. This can also be seen on an SEM which is equipped with scanning coils in the gun. We provide this kind of "source imaging" feature on our Personal SEM(TM) and we train our users to use it to adjust the filament drive so as to achieve the most compact source emission pattern. With this tool, it is easy to demonstrate that if you mis-adjust the bias resistor, you can observe an emission "saturation" effect (emission current stops increasing as the filament temperature is increased) when the emission pattern is actually far from optimal. Typically, this occurs when the bias resistor is set too small and thus one has selected too large a region of the filament tip for emission.
So I hope that I have now clearly made my point that "saturating the gun" is just a technique we use to achieve an otherwise desireable result -- putting the microscope into a stable current configuration which we have set up to also provide the best imaging conditions. The notion of "saturation" per se is really without meaning for electron microscope optics. It's rather unfortunate that this term entered our vocabulary since it is quite misleading (maybe we should call it "stabilization"). But that is the term we are stuck with, it seems (kind of like my other pet peeve: Energy "Dispersive" Spectroscopy). =================
Steve's comments about the role of filament depth, kilovoltage, and anode spacing are generally sound advice, but when he speaks about "efficiency", he starts to blur the issue. As already noted, one wants to get the electrons away from the cathode as rapidly as possible so that there is a minimal space-charge effect. This is accomplished by maintaining a high field gradient in the gun and this can be accomplished by upping the acclerating voltage, moving the anode closer to the cathode, or moving the cathode closer to the wehnelt (you could also insert a separate "extraction" electrode in the gun as has been done on occassion [Yamazaki et al, 1984] and is effectively what is done in field emission guns). Note, however, that one's objective in doing this is not to generate more emission as such, but rather to improve the quality of the beam by overcoming the space charge "Boersch effect". In other words, you don't necessarily need more electrons, but more electrons going in the right direction.
When Steve speaks about the "efficiency of the gun", the implication is that the goal is to produce lots of electrons (emission). I doubt that this was his actual intent, since that would badly miss the point. When we speak of the "efficiency" of a gun, we are properly speaking about the ratio of "brightness" to total emission. The differentiation between emission and brightness seems to confuse a lot of people. I like to illustrate the difference by posing the following problem: suppose I take two people and offer a prize to the individual who can use a water hose to spray the maximum amount of water through a knothole 100 feet away. I then tell each contestant that they have a choice of two hoses, one with a discharge rate of 10 gallons per minute and the other with a discharge rate of 1 gallon per minute. The "foolish" contestant immediately opts for the higher flow rate, and quickly reliazes his error when I then hand him a hose which has a nozzle producing a fan-shaped stream -- although the flow rate is high, only a small fraction of it can be directed through the knothole. The more thoughtful contestant asks to see the shapes of the streams produced by the two hoses and chooses the lower-flow hose which happens to have a narrowly collimated stream -- and wins the contest. The point, of course, is that it is not how many electrons the gun produces, but rather how many can be directed through the electron optical system. We call this quality "brightness" -- it is the combined spatial and angular density of the beam. An ideally "bright" beam would be a non-divergent "pencil" of electrons which is very dense (the water hose analogy breaks down here since water isn't very compressible). Brightness is the quality which we should worry about (rather than emission current) because it dictates the quality of the image spot which we can form on the sample.
There is a well-known theory due to Langmuir in which he establishes the maximum brightness value for a thermionic (e.g., tungsten or LaB6) cathode. The variables are the emissivity of the cathode material, the accelerating voltage, and the temperature of the filament. So how does one design a gun to achieve this maximum brightness? Haine answered this question in that same paper referenced above. The answer is that over quite a wide range, you can achieve the Langmuir limit with a variety of gun designs. In particular, you can vary the size of the wehnelt opening and the spacing of the filament behind the wehnelt over wide ranges and still approach the Langmuir limit so long as you adjust the biasing appropriately. What does change, however, is the "efficiency" of the gun. By efficiency, he means the ratio of brightness to total emission. Returning to the "water through a knothole" example, you can imagine that the shapes of the two hose streams are such that either hose can deliver the absolutely maximum volume of water through the distant knothole at the operating pressure of the supply line (like achieving the Langmuir limit for a given beam energy with two different gun designs). But in this case, the lower volume hose does the job with much higher efficiency, and normally would be the preferred solution (unless we enjoy wasting water).
If the wehnelt opening is large and/or the filament set well back, then you will be able to achieve the Langmuir limit only by operating with a rather high emission current. Since there is really no advantage to having all of that wasted emission current (and some practical disadvantages) it is preferable to design the gun for high efficiency. SEMs today are generally designed for more efficient cathode operation than was the case in the past. By way of comparison, a 1970's vintage ETEC operated with a 200 microamp emission current, whereas most modern SEMs operate at 50 microamps or less. The highest efficiency is obtained by placing the filament tip close behind a small wehnelt aperture. But the closer the spacing, the higher the required bias and this can lead to unstable operation if pressed too far. A very small aperture makes precise alignment of the gun particularly critical. Thus, there are tradeoffs and the gun designer will try to weigh a whole lot of factors. The end user doesn't need to worry about these things. If the design and setup is proper, setting the filament to "saturation" will approximate the Langmuir brightness limit. =====================
I do have to take strong exception to Steve's statement that: "if you do not have enough [electrons] to start with you soon run out, hence poor signal levels and low magnification microscopy!" I'm sure that this isn't precisely what Steve meant to say, but I'm going to pick on the statement anyway, since a lot of people seem to think it is true. The important question isn't how MANY electrons you have to start with, but rather, how they are DISTRIBUTED in the beam. Lose sight of that fact and you can end up drawing some rather odd conclusions. For example, would you conclude that you are going to get better imaging with an old Etec with its 200 microamp emission current, or a modern cold field emitter where the emission is orders of magnitude smaller? Clearly, emission QUALITY is vastly more important than emission QUANTITY. I make this point emphatically because so many people seem to feel that anything they do to their microscope which increases the emission will somehow increase the image quality. Sometimes it is just the opposite. To make an extreme example, if you want tons of emission current, just drill out the hole in the end of your wehnelt to a half inch diameter -- LOTS of emission, but TERRIBLE imaging. =====================
This has gotten a bit off topic from the simple question of "filament life", and I hope that anyone who has stayed with me till here can pardon me for such a lengthy reply (but I did warn you). I do hope this lengthy diatribe helps to illustrate that though filament spacing and the like do have a relationship to gun performance and ultimately to filament life, these are actually secondary effects which represent design and/or operational decisions rather than fundamental optical considerations.
} } } I read with interest Fred's note but fear he has missed one important } feature of saturation? } } Saturation occurs when the aperture in the front of the cathode is filled } with electrons, generate more and they are unable to pass through therefore } no change in emission current. } } What affects this aperture? The physical size of the aperture and the bias } field. Apply more bias and the effective aperture size is reduced, } therefore the number of electrons needed to fill the aperture is reduced, } therefore the amount of heat required to produce the required number of } electrons for saturation is also reduced. Hence apply more bias you need } less heat you get a longer filament life as heat (oxidation and } evaporation) is the determining factor. } } With regard to the position of the filament the further it is away from the } cathode aperture the greater the affect of the bias field. Hence a long } filament to aperture distance is effectively increasing the bias field } requiring less electrons for saturation therefore less heat and a longer } filament life. } } The best instrument to use to see this in action is the TEM where we may } visualise the virtual source and may see the effects of bias on saturation. } } When we lower the kV the gun becomes less efficient due to the anode to } cathode distance being too great for the applied voltage and the resulting } field being less efficient in helping to create the virtual source. The } source is also affected by the higher residual bias field in most SEM which } throttles the cathode aperture still further. If you lower the kV and want } maximum efficiency you should move the filament forward to overcome the } bias field and raise the anode to increase the efficiency of the } anode-cathode field. } } Hope this helps, the truth is that even with field emission if you do not } start with a suitable emission current the potential for good quality } images is drastically reduced. Remember everything we do once the beam has } left the electron gun throws electrons away, if you do not have enough to } start with you soon run out, hence poor signal levels and low magnification } microscopy! } } The SEM is an exciting SCIENTIFIC INSTRUMENT, understand it and it becomes } an amazing tool, mis understand and it becomes a super light microscope; } what a waste? } } Steve Chapman } } Senior Consultant E.M. } Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. } Tel & Fax 44 (0)1844 353161 } Web Site - http://ourworld.compuserve.com/homepages/protrain } For Consultancy and Courses in Electron Microscopy World Wide }
The problem that you outline is very common when people change instrument= s. Zeiss using a SE detector gives satisfactory BSE images, LEO using a BSE=
detector you do not like the image as much! =
Using the so called SE detector for BSE collection you may achieve a good=
signal but only with a large surface area detector. This signal also contains a shadow effect due to the specimen-detector geometry. Trying t= o use the LEO in the same way as the Zeiss will not work as the LEO SE detector has too small a surface area for this style of operation.
The LEO BSE detector being directly above the specimen will be free from shadows, unless you set it up in the TOPO mode, or move the BSE detector off axis sufficient to clear the beam path. Detector variations will be available to you in the DETECTOR menu under SELECT SIGNAL, either do not switch all the segments on or switch to the TOPO mode. Try operating at different WD to optimise the LEO specimen-detector geometry?
Another problem, as you work at very low magnifications, may be you do no= t have enough signal, out up the probe current? Also the bigger the spot t= he better under low mag BSE circumstances. Do not tell anyone I said this b= ut you may find you get a better image by running at the false peak often called the first peak of saturation. This gives you a bigger source and hence a larger final probe size.
Good luck
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Regarding improving the performance of your Amray SEM the same rules appl= y as I passed on to Paula. (Sorry I do not have specific instructions for a=
1600 only an 1800 and I do not know how they compare?)
1. You need current - through higher emission levels
2. You need to saturate and align correctly to aid the above
3. You need to run in a minimum aberration condition - short working=
distance
4. Plus the following which are Basic SEM Adjustment Procedures
Image is Too Noisy?
1 Spot size is too small? 2 Emission current is too low? Variable aperture is too small? 4 Specimen position is not ideal - too far from the detector or tilted away from the =
detector? 5 High voltage is too low?
Resolution is Poor?
1 Spot size too large? 2 Working distance too long? 3 Final Aperture too large? 4 Emission current too low or saturation and alignment poor? 5 High voltage too low? 6 Image does not contain high resolution information? 7 Too much backscatter in the image (kV too high or specimen tilted=
towards the =
detector)?
Presentation of the Image is Poor?
1 Spot size incorrect, too small or too large? 2 Would tilt help present the information? 3 Have you the ideal WD? 4 Have you the correct kV for the job? 5 Have you the correct aperture (small) to obtain a good depth of field, do you need to =
move the specimen further away from the final lens? 6 Are you seeing a mixed SE/BSE image when you need a BSE or SE image. Lower =
the kV for the latter raise the kV for the former.
Hope this helps?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
----------------------------------------------------------------- PHILADELPHIA SOCIETY FOR MICROSCOPY ----------------------------------------------------------------- AN AFFILIATE OF THE MICROSCOPY SOCIETY OF AMERICA and THE MICROBEAM ANALYSIS SOCIETY -----------------------------------------------------------------
MEETING NOTICE - SPECIAL EVENT: Thursday, May 13, 1999
METALLOGRAPHY AND THE STUDY OF THE JAPANESE SWORD
Michael R.Notis, Lehigh University, Bethlehem, PA 18015
To be presented at The Egyptian Gallery, University of Pennsylvania Museum of Archaeology and Anthropology Reception at 5:30 followed by Dinner at 6:30 and the talk at 7:30
This meeting is intended to be of interest to both microscopists and a more general audience. We encourage you to bring guests and spouses. Cosponsor: the Museum Applied Science Center for Archaeology (MASCA), University of Pennsylvania Museum of Archaeology and Anthropology. ----------------------------------------------------------------- Meeting Details
Location: The Egyptian Gallery, University of Pennsylvania Museum of Archaeology and Anthropology (map and directions enclosed). Parking is available behind the LRSM after 5:00 PM and in the parking lots indicated on the attached map.Cost of Dinner: Members and members spouses $20.00, Students $15,Non-Members $30. Schedule: 5:30 Social hour hosted by our meeting sponsor. Beer, wine, soda and munchies 6:30 Dinner Salad- Watercress, Belgian endive, apples, alfalfa sprouts and raspberry vinaigrette. Entree- Seared breast of chicken served with porcini and shiitake mushrooms, onions, scallions, ginger and garlic topped with a black bean sauce. Vegetarian entrees available upon request. Dessert- Tiramisu 7:30 TALK ----------------------------------------------------------------- Reservations
By E-Mail (preferred): Send your name and affiliation to PSM-RESERVATIONS-at-INAME.COM By Phone: Call Dr. John Reffner, 215-619-5283 I will reply to confirm all email reservations received. DEADLINE for RESERVATIONS is Noon, Monday May 10th. ----------------------------------------------------------------- Abstract
The ancient Japanese sword is a marvel of empirical technology and has become a well known example of the beauty of metals as an art form. The manufacturing process (religious ritual) through which the sword is made, the heat treatment it receives and the microstructure that is developed will all be described. By the late 19th and into the early 20th century, the first Japanese metallurgists were studying modern advances in metallographic techniques. Among them were K.Tawara and his student H.Tanimura. This connection between sword study and metallography led to interactions between Tanimura and C.S.Smith, whose own metallographic studies of ancient metal objects, including the Japanese sword, are well known. A group or important swords originally studied by Cyril Stanley Smith using light optical microscopy have been reexamined using scanning electron microscopy and energy dispersive x-ray spectroscopy in order to better examine the microstructure and to perform microchemical analysis on the slag inclusions. The use of these newer methods for analysis demonstrates the power of microchemical analysis in combination with metallographic examination to extend our knowledge concerning these ancient swords and their fabrication. A VIDEO OF THE SWORD MAKING PROCESS WILL BE SHOWN
1. Take I-76 (Schuylkill Expressway) to the South Street; exit (left lane exit). From the west, it's the exit after 30th Street; from the east, the exit after University Avenue. Turn west (right from the west; left from the east) at the light at the top of the exit ramp. The Museum is on theleft past the next light. 2. From the PA Turnpike: Valley Forge Exit to I-76 (Schuylkill Expressway, east). Then follow directions in #1. 3. From the New Jersey Turnpike: Take Exit 3 to Walt Whitman Bridge. Once across, follow signs to I-76 (Schuylkill Expressway, west). Then follow directions in #1. 4. From I-95 North: Exit at Vine Street. Take Vine Street west to I-76. Stay in left lane. Take exit marked "To Airport" (a left U-turn). Take second exit (South Street). Then follow directions in #1. 5. From I-95 South: Just past Philadelphia Airport, follow signs toI-76. Once on I-76 follow directions in #1.
Parking lots (pay) are available behind the Museum, one block over the bridge off Convention Blvd (turn left). The Museum is on South Street, with its main entrance at the corner of 33rd and Spruce Streets. In addition, parking will be available at the usual location, and free parking for about 15 cars is available at the Sharpe Circle. Information on the Egyptian Gallery is available at
DENTON VACUUM- Three decades of manufacturing vacuum equipment for the preparation of specimens for electron microscopy.
PHILLIPS ELECTRON OPTICS, Phillips Electron Optics, part of FEI Company,is celebrating the 50th anniversary of the first commercial EM-100 TEM in 1949. We are setting the pace in electron microscopy for the new century with our latest TECNAI family of TEM's along with the XL Series SEM's and focused ion beam (FIB) systems.
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The Microscopy Society of America is also providing financial support for this meeting.
} You can mimic this analog technique by using a very large blurring kernal } on the original and subtracting the blurred image from the original image. } If necessary, you can multiply the blurred image by a constant to adjust } the final intensity. You can also define a custom kernel to get something } large enough to do what you want. This should help even out the background } in your image. } } Henk } } At 03:17 PM 5/5/99 +0100, you wrote: } } } } } } And now the question: one has a scanned or digitally acquired micrograph. } } The background is a gently varying grey, with rather low contrast local } } features (blobs, bacteria, or whatever). The task is to Fourier } } transform, then filter out the lowest frequencies with a high-pass filter, } } leaving one with the features on a uniform grey background. One can then } } increase the contrast to make the features more prominent for printed } } reproduction. Is there software (Win 95) that can do this? (preferably } } inexpensive, though we could make arrangements to use plugins for Adobe } } Photoshop elsewhere, if necessary).
You could look for the key-words 'median filter', 'blurring', 'non-linear filter'. NIH-Image might well have such filters. If not there are other free programs, such as Osiris: http://www.expasy.ch/www/UIN/html1/projects/osiris/osiris.html Image Tool: http://ddsdx.uthscsa.edu/dig/itdesc.html
You can find more software at the shareware-site: http://www.nsctoronto.com/nissei-sangyo/ftpsw.html
To use the median-filter do as Hendrik Colijn describes above. If the filter kernel is twice as big as the largest object in your image the filtered image will just show the smooth background. Subtract the filtered image (maybe scaled) from the original to get the objets on a flat background.
-- Philip Koeck Karolinska Institutet Dept. of Bioscience Novum S-14157 Huddinge Sweden Tel.: +46-8-608 91 86 Fax.: +46-8-608 92 90 Email: Philip.Koeck-at-csb.ki.se http://www_scem.csb.ki.se/pages/philip.html
There is one ethanol/ethylenglycol-based fixative on the market=20 (Kryofix/Merck No. 5211) which was developped and used as a substitut= e of =20 formaldehyde ("formalin") for routine preparation in a pathology/hist= ology lab=20 of M.E. Boon in Leyden (Belgium). Best results were obtained in=20 combination with microwave irradiation and in immunostaining. See= =20 references in the "Microwave cookbook of pathology. the art of micros= copic=20 visualization" from M.E. Boon and L.P. Kok, Coulomb Press Leyden. 3rd= =20 edition 1992. Best regards Bernward Laube University of Bielefeld Faculty of Biology Department Plant Morphology and Cell Ultrastructure Universit=84tsstrasse 25 Germany 33615 Bielefeld phone: +49 521 1065592 fax: +49 521 1066039 e-mail: b.laube-at-biologie.uni-bielefeld.de http://www.uni-bielefeld.de/biologie/Pflanzenmorphologie
Please unsubscribe the following id: srinivasan-at-alpha.caer.uky.edu Sincerely Ram Srinivasan Center for Applied Energy Research 2540 Research Park Drive Lexington, KY 40511 (606) 257 - 9695 (Off) (606) 223 - 7378 (Home)
There are several solutions out there for capture of video from DV format cameras. They include the following:
Bravado DV2000 DPS Spark Miro DV300 Fast DV Master Canopus DVRex-M1 Radius EditDV
All of these are video capture cards that usually include some video editing software designed to work with the card. The last suggestion from Radius has included within the package a program called MotoDV for video capture and I believe it will capture still frames. A lot depends upon how much you want to spend, what computer platform you are using, and the type of hard drive it writes to. For a 2:1 compression ratio (good quality video) you need a drive with a transfer rate of at least 10 Megabytes/sec. Your frame size will be about 333 kilobytes and you will need 1.67 Gigabytes of space for each minute of video. Not sure what your application requires for the still frames but there quality (such as for printing) may not be great.
http://www.videoguys.com/jump.htm
http://www.videotexsystems.com/
Try these WEB sites for more information. Hope this is helpful to you.
Regards
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
Do beware that the typical IDE HDD with an advertized data transfer rate of 33 Meg/sec cannot usually sustain that rate. For example, My WD 8G (UDMA) drive benchmarks in the area of 4 Meg/sec sustained. Depends on the drive, interface type, etc.
There are several solutions out there for capture of video from DV format cameras. They include the following:
Bravado DV2000 DPS Spark Miro DV300 Fast DV Master Canopus DVRex-M1 Radius EditDV
All of these are video capture cards that usually include some video editing software designed to work with the card. The last suggestion from Radius has included within the package a program called MotoDV for video capture and I believe it will capture still frames. A lot depends upon how much you want to spend, what computer platform you are using, and the type of hard drive it writes to. For a 2:1 compression ratio (good quality video) you need a drive with a transfer rate of at least 10 Megabytes/sec. Your frame size will be about 333 kilobytes and you will need 1.67 Gigabytes of space for each minute of video. Not sure what your application requires for the still frames but there quality (such as for printing) may not be great.
http://www.videoguys.com/jump.htm
http://www.videotexsystems.com/
Try these WEB sites for more information. Hope this is helpful to you.
Regards
Roy Beavers Southern Methodist University Dept. of Geological Sciences Electron Microprobe Lab P.O. Box 750395 Dallas, Tx 75275 voice: 214-768-2756 fax: 214-768-2701 E-mail: rbeavers-at-mail.smu.edu
[frequent skips; } delineates Fred's post, and } } delineates Steve's]
} Consistent with that sentiment, I am } going to launch into a fairly lengthy discussion of the phenomenon that } we know as "saturation", since it is my experience that it is widely } misunderstood.
} } Saturation occurs when the aperture in the front of the cathode is filled } } with electrons, generate more and they are unable to pass through } } therefore no change in emission current. } } } So I will stand by my original assertion that the } effect that we call "saturation" is simply a consequence of the common } bias-resistor circuit, nothing more. } } Haine in a 1952 paper on page 45 states: "...With this type of biasing, } an electron gun exhibits the property, frequently termed 'saturation,' } whereby the current rises with temperature to a limiting value beyond } which increase in temperature produces little increase in beam current." } } To drive the point home: the effect which } microscopists call "saturation" is nothing more than reaching the } operating point in the emission current stabilization circuit } established by the bias resistor -- there is no profound electron } optical effect involved. Certainly it is not a matter of the electrons } having "filled" a virtual "aperture" established by the bias field. } } What you actually want to do, of } course, is set the bias so that just the tip of the filament is able to } emit. In this "independent bias" configuration, changing the filament } temperature has no effect on the bias, and thus no effect on the } position of the emitting region.
} For another thing, the relationship } between filament temperature and emission current is different above and } below the "saturation" knee.
} But, if you have any sense at all, you select the bias } resistor value such that the "saturation" knee occurs at a filament } temperature which gives both a nice compact emission pattern AND an } acceptable filament lfe.
} As Steve has pointed out, the effect on the emission pattern of changing } the bias can be readily observed on a TEM. } } So I hope that I have now clearly made my point that "saturating the } gun" is just a technique we use to achieve an otherwise desireable } result -- putting the microscope into a stable current configuration } which we have set up to also provide the best imaging conditions. The } notion of "saturation" per se is really without meaning for electron } microscope optics.
Dear Fred, I realize that the situation is considerably different for the TEM from that for the SEM; however, what I have always heard and observed is that for low filament currents electron emission occurs only from some regions of the filament tip. This can be seen by imaging the filament at crossover, where it appears as an annulus. As the filament current is raised, additional regions of the tip begin to emit electrons, and the image of the filament becomes more uniform and smaller. As the filament current reaches a value such that all regions of the tip emit, the image has no dark regions. This is what I have called "saturation"; it makes sense from the standpoint that emission cannot occur from more than the complete tip area and, therefore, cannot increase with further increases in filament current. AFAIK, this has nothing to do with space charge, current stabilization, virtual apertures, or bias resistors, but with the temperature, geometry and work function of the tip. Are there two uses of "saturation"? Yours, Bill Tivol
First I would like to thank those who responeded to my earlier inquiry about the Lynx tissue processor. I have since received info on the RMC EMP 5160 tissue processor and would like any info anyone can give me on this processor, be it good or bad.
Instead of writing directly to disk, you can always write to a RAM buffer first. For (uncompressed) video you have 30fps and each frame is about 1/3 MB (640x480), so that's only 10MB/s. You can buy RAM boxes with multiple-GByte capacity -- Texas memory has such a box with 12GB of RAM and a 1.2GB/s bandwidth that would buffer about 20 minutes worth of (uncompressed) video -- you could then "drain" this memory to your IDE HDD at 4MB/s. At the high end, you could buy a box with 128GB to hold over 3 hours of video!
See: http://www.texmemsys.com/samsys.htm
-Mike O'Keefe
"White, Woody N" wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Do beware that the typical IDE HDD with an advertized data transfer rate of } 33 } Meg/sec cannot usually sustain that rate. For example, My WD 8G (UDMA) } drive } benchmarks in the area of 4 Meg/sec sustained. Depends on the drive, } interface } type, etc. } } Woody White
I have been working for a long time in the field of inorganic materials. Now I am getting into biomaterials and my first task is to determine porosity of a bone, I think I need some help. I guess that if I record images of the surface of the bone and apply sequential sectioning and image recording, I will be able to reconstruct the 3d characteristics of the object. I wonder if there is any software available to do the reconstruction or maybe someone can suggest some references where I could get started.
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I have been off-line for some time and may have missed relevant postings.
I have been asked to enquire about collecting digitised TEM and STEM images from the above microscope in order to improve the facility. The instrument dates from 1981. We are a little behind the times in this area so any information would be appreciated. Maybe pointers to the archives? Clues as to when this may have already been discussed !
Thanks in advance
Keith Ryan Marine Biological Association of the UK Citadel Hill Plymouth Devon PL9 9DR England
Firstly I have to say that when talking about improving performance I tak= e it for granted that people clean the column and gun components. My objec= t is always to move on from this point.
Now when we use a SEM at low magnifications ( {3,000X) the size of the pro= be encourages BSE to totally dominate the image particularly when using in excess of 10kV. This means that the SE detector collects a signal made u= p of direct SE, plus SE which are converted from BSE that hit the lens etc,=
plus line of site BSE (well documented in the literature).
When we view an image which contains high levels of SE it tends to be rather flat and devoid of high contrasts, the best sign of a BSE contribution is the formation of shadows. Lowering the kV we obtain an image that displays more of its SE content and less of the BSE content so=
this is the best way to analise the imaged information. =
To fully understand an unknown specimen use this procedure
1. Start at low kV ( {2kV) when you see the true specimen surface 2. Increase the kV until you see an change in the image form, now yo= u have increased the reaction volume to such an extent that the BSE start t= o dominate and you are now bringing in sub surface detail. 3. The higher the kV you use the greater the sub surface (BSE) contribution. 4. If as you go up in kV the image becomes more exciting (more detail?) this means that there is more sub surface data coming out by way=
of BSE. 5. If you go up in kV and the image becomes flat this means that the=
sub surface zones do not have much detail and the result will is a softening of the image.
(The ideal test specimen to demonstrate the above in action is a TEM specimen. I use a SPI carbon grating replica which I tack to a stub with=
the carbon side up. Take a look at different kV and blow your mind as yo= u see so much information goes missing when the kV goes up. Sure it is a very thin specimen but it is a great demonstration.)
It is for the above reasons that I frown upon those who insist on using 20kV plus for imaging, because most of the time their information is not = of the surface but of the sub surface. This is fine if you know what is happening, but most people do not understand the specimen-beam reactions and the way they relate to specimen-detector geometry.
Hope this helps?
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Whilst I think people should change their scintillator every few years (y= ou clean your glasses don't you?) I have to say that, when experimenting wit= h signal and different types of scintillator material many years ago, the results did not push me too hard in this direction.
We found that a new scintillator gave a reading which very quickly fell a= nd then took an age (years) to drop to a level that one would consider was unacceptable.
Another area where we have had fun is that of photomultipliers, when the BSE signal from its detector is far stronger than the Everhart-Thornley detector its about time you changed it!
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain
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___________________________________________________________________________ Pearson Coal Petrography Inc., Pearson & Associates Ltd., 16130 Van Drunen Road, 4277 Houlihan Place, South Holland, Victoria, Illinois 60473 British Columbia V8N3T2 United States Canada
For program, flyers, abstract submission, and registration go to: http://online.sfsu.edu/~camicro/
2nd Announcement and Call for Papers 5th California Microscopy Colloquium The California State University & Northern California Society for Microscopy
Saturday, October 2, 1999 Business meeting for CSU delegates, Friday, October 1, 1999 Seven Hills Conference Center San Francisco State University
Early Registration Due August 2, 1999 Abstracts Due August 2, 1999 Abstracts will be published in the journal Microscopy Research and Technique
All fields of light and electron microscopy. Participants include scientists and students from academia and industry. Both platform and poster presentations are invited. Student posters and presentations are strongly encouraged with award of meritorious original papers/posters.
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Bill Tivol makes a good point when he asks whether there are two different meanings attached to the term "saturation" (see appended quote at end).
As Bill correctly observes, there is a real physical effect where the beam emission pattern can be observed to transition from a diffuse annulus (what we used to call a "smoke ring") into a nice compact disk. This can easily be observed on a TEM (or so I'm told -- I'm not a TEM guy and have only seen this illustrated in publications). But a phenomenon which looks very much the same can be observed on a SEM equipped with gun scanning coils -- I happen to like this feature a lot and have designed it into our SEM since I find it the most reliable way of optimizing the gun. (By the way, I am unaware of any practical differences between SEM guns and TEM guns other than is related to the differences required by the TEM's higher operating voltage -- the basic optical principles are the same.)
When you do look at the emission pattern in this way, as Bill has noted, you can clearly see what it is you are trying to achieve. When "undersaturated", the beam profile is "empty" in the center. If you look at a "line scan" through the pattern, the profile looks a bit like the letter "M" -- this is clearly not what you want. As the biasing is increased (accomplished in the self-biased gun by either increasing the filament temperature or by increasing the bias resistor) the pattern collapses into a disk. In cross section, this distribution is high in the center and falls off steeply to the edges -- looking very much like a gaussian distribution (though I have no proof that is actually its mathematical shape). This is the condition you are trying to achieve, and calling this "saturation" makes a lot of sense. So Bill is perfectly correct in noting that there is a second meaning to the term "saturation" which has real validity for tuning of the gun.
My problem with the term "saturation" is that it has been used to refer to at least three different phenomena: 1. The idea of saturating the emission "phase space" with electrons such that no more can be crammed into the beam (the "high perveance" condition I referred to yesterday) -- as near as I can figure out, this is what people thought was happening when they originally used the term "saturation" in electron microscopy since it does occur with other types of electron guns. 2. The coalescing of the emission into a compact pattern as the emission becomes restricted to the tip of the filament. 3. The "knee" in the emission curve in an auto-biased gun -- the point where measured emission current stops increasing as filament temperature is increased
The first effect just doesn't apply to an electron microscope, and the third effect is simply the manifestation of the auto-bias circuit (these are the two points I argued 'ad nauseum' in yesterday's posting). The second effect is both real, and desirable -- this is what we are trying to achieve when a microscopist "saturates" the gun. I don't have any trouble with calling this "saturation" (and usually do so myself) since it is a pretty good description of what one is visually observing. The problem is, unless you have a TEM, or a SEM equipped with "source imaging" (which is apparently not all that common), people tend to think that the third effect is due to one of the first two -- and this is just not so. This mistaken notion is what I was laboring to correct. But it was never my intention to dispute the fact that something very useful happens to the beam shape when you correctly adjust the biasing (effect #2). If your microscope is setup correctly, the auto-bias "knee" (effect #3) will occur in the vicinity of the compact source distribution (#2) and using the "knee" to saturate works just fine. But if one starts arbitrarily tinkering with filament spacing and the like so as to increase emission, extend filament life, etc., without understanding what is happening, effect #3 may not match effect #2 anymore and when the operator thinks that he/she has optimized the gun by "saturating the filament" via the emission curve knee, the emission pattern may actually be non-optimal.
To address Bill's remarks about the mechanism which results in this localization of the beam in the emission image, I'm now going to restrict myself to discussing effect #2 only -- and I'll call it 'saturation" since that's what we're used to.
What causes this 'saturation' of the emission distribution to occur? Or equivalently, why does one get the hollow-centered "smoke ring" type of pattern when the filament is "undersaturated"? Bill postulates that this is because electrons are only emitted from the periphery of the circular emitting region on the filament tip (i.e., the region selected by the biasing). I don't think this is quite correct since it is energetically possible for electrons to be emitted from anywhere within the "zero equipotential" region established by the wehnelt bias. Instead, the answer is related to the fact that biasing the wehnelt not only establishes the emitting region on the filament tip, but also creates an electrostatic lens which focuses the beam (I've ignored the focusing aspect thus far in the discussion since things were complicated enough without getting into this -- and my posts were already plenty long!).
But focusing does need to be introduced if we are going to understand the emission pattern since we need to be aware that what we are seeing in the "filament image" is not an image of the filament per se, but rather, an image of the "crossover" distribution, which is the point where the focused electrons from the filament converge to their smallest cross-section. (Someone may want to argue with me about this, since if you get into really undersaturated conditions, you can actually see structure on the filament tip. However, this is because the "crossover" is in effect a spatial image of the filament tip under these conditions. In the optical sense, the "object" you are looking at in a "filament image" is this crossover image, not the physical tip as such.) So just because we don't see any electrons in the center of this emission pattern does not mean that the filament is not emitting from its center -- it means that the focusing action is such that the center of the crossover pattern isn't getting electrons directed into it.
I again like to use a garden hose analogy to describe what is happening here. Imagine how you adjust the nozzle on your garden hose to move from a fan-shaped (open-centered) distribution to a compact stream. I visualize the bias on the wehnelt shaping the beam in a rather analogous fashion. The emission distribution image you observe is thus like observing the cross-section of the water stream at some distance from the nozzle. If you think about trying to direct the tightest possible stream of electrons down the optics of the column, you can then see why "saturating" the beam (in sense #2, of course) gives you the best imaging conditions. (I hasten to add the disclaimer that I NEVER want to be accused of claiming that a garden hose is a perfectly accurate model of gun dynamics -- just a useful visual metaphor for some phenomena.)
Two other related topics suggest themselves. One is the way that this "garden hose" model explains the phenomenon of "false peaks" which you obtain with a misaligned gun when you are trying to "saturate" by watching the specimen current. The second relates to the question of why the "compact distribution" condition is optimized when the emitting region is restricted to the filament's tip. However, I don't want to engage in another marathon post like yesterday's (the Bandwidth Conservation League will probably censure me as it is) and I also have no idea whether anyone but myself is interested in such arcane topics. So I will (for once) refrain.
Fred Schamber RJ Lee Instruments
====================== Bill Tivol wrote:
} Dear Fred, } I realize that the situation is considerably different for the TEM } from that for the SEM; however, what I have always heard and observed is } that for low filament currents electron emission occurs only from some } regions of the filament tip. This can be seen by imaging the filament at } crossover, where it appears as an annulus. As the filament current is } raised, additional regions of the tip begin to emit electrons, and the } image of the filament becomes more uniform and smaller. As the filament } current reaches a value such that all regions of the tip emit, the image } has no dark regions. This is what I have called "saturation"; it makes } sense from the standpoint that emission cannot occur from more than the } complete tip area and, therefore, cannot increase with further increases } in filament current. AFAIK, this has nothing to do with space charge, } current stabilization, virtual apertures, or bias resistors, but with } the temperature, geometry and work function of the tip. Are there two } uses of "saturation"? } Yours, } Bill Tivol }
Barry, Lilith wrote: } } Thank you for responding to my question on tissue culture background. I } don't think the background is because the fixation or the immuno method } because I routinely use the same method on brain sections and do not get the } same background. It isn't the coverslip autofluorescing either because one } can see beautiful labeled structures in the cells except on the controls. So } it has to be something specific about cortical cultures and the strept or } neutravidin-biotin method. Do you have any other thoughts? } Lilith } ------------------------------------------------------------------------ } Lilith Ohannessian-Barry } National Research Council } Institute of Biological Sciences } CANADA } Tel;613-993-6460 } Fax;613-941-4475 } e-mail; lilith.barry-at-nrc.ca
Hi Lilith, Have you ruled out the endogenous biotin? Either try a biotin block prior to your staining or try another detection method, which does not utilize biotin. Katri
Is there any significant differences in SEM filament saturation respective to W, LaB6 or field emission cathodes? If so, what are they and how should one treat these different electron sources?
TONS, yes the difference between W, LaB6 and an FEG "saturation" (lets s= ay source set up) is LARGE.
For your application LaB6 saturation should be carried out when the virtu= al source is being visualised as there are a number of peaks that you may us= e depending upon the work you wish to carry out, high brightness, medium resolution and high resolution. =
May I suggest you contact the supplier of your LaB6 cathodes and they are=
sure to be able to give you a sheet relating to saturation and good housekeeping for LaB6, you just cannot treat these sources like W, they a= re very sensitive! =
A LaB6 tip will become damaged by discharge (a W will usually not unless=
at TEM voltages) such that it may emit from more than one area. In order=
to prevent this you should run up the kV and filament very very slowly. =
This allows the gas emitted through these actions to be removed before th= e gun environment becomes suitable for discharge. W filaments will take al= l that you throw at them LaB6 I say again are very very sensitive but boy d= o you get some gains when they are up and running? Higher brightness less aberration therefore higher resolution and particularly at low kV.
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Scanners Corporation has an opening for a Senior SEM Field Service Engineer. This position is for the Eastern U.S. region.
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Scanners Corporation is one of the oldest and largest third party SEM service companies in the U.S.. We offer a full benefit package, above industry average salaries and commission. Please email or call for additional details. All replies will be kept in the strictest of confidence.
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I think a good starting-point could be the Reconstruction-Homepage with several references, software descriptions and links: http://biocomp.stanford.edu/3dreconstruction/index.html For basic information, maybe the following article is of help: http://www.videomicroscopy.com/Tutorials/3D%20Reconstruction/October%20hows% 20that%20work.htm I guess that for your task volume-renderer or ray-tracer software will be a better choice than surface-renderer. Some of the volume based programs are free, e.g. NIH-image for Mac (with integrated 3D functions) http://rsb.info.nih.gov/nih-image/ There is also a Windows version, called Scion-Image http://www.scioncorp.com/frames/fr_download_now.htm However, with such a complex structure like bone I would think about a way to use a confocal laser scanning microscope. This would avoid the problem of image alignement and you can use the integrated 3D-functions of the cLSM software. Maybe someone on the list does know a protocol for thick sectioning of bones and visualization with the LSM?
Best regards
Jens
} Hi everyone: } } I have been working for a long time in the field of inorganic materials. Now } I am getting into biomaterials and my first task is to determine porosity of } a bone, I think I need some help. I guess that if I record images of the } surface of the bone and apply sequential sectioning and image recording, I } will be able to reconstruct the 3d characteristics of the object. I wonder } if there is any software available to do the reconstruction or maybe someone } can suggest some references where I could get started. } } Thanks in advance } } } Hector Calderon
------------------------------------------------------------------- Dr. Jens Buecking Tel. +49-(0)421-218 3745 University of Bremen Fax. +49-(0)421-218 4620 Dep. of Biology Email jbueck-at-biologie.uni-bremen.de Leobener Str. - NW2 URL http://www1.uni-bremen.de/~jbueck/ D- 28359 Bremen (AG Prof. Dr. H. Witte) GERMANY
2tes Acarologisches Kolloquium in Bremen, 14.-16.10.99 http://www-user.uni-bremen.de/~acari/ -------------------------------------------------------------------
I've received a few replies to my inquiry about TV rate performance of solid-state vs. scintillator BSE detectors. Thank you.
It seems that the p-n type solid state BSE detectors are not quite up to the performance at TV rates of the scintillator types, but solid-state systems such as those provided by GW Electronics do provide good TV rate performance.
Bruce E. Brinson has informed me that micro-channel plate type BSE detectors also perform adequately at TV rate and much better at that rate than some p-n solid state type detectors he has used.
In my case, I was attempting to use an "in-my-lab" modified Robinson BSE detector system. My TV-rate performance was not good enough. The light pipe shape that I prepared for my ARL SEMQ's BSE system is far from optimum because of little available space for an "overhead" scintillator and a slightly remote location needed for the PMT.
Another modification performed a few days ago has now given good TV rate performance. A simple idea. Tricky to execute. I cored out the lens bore area of the plastic scintillator of the original and replaced that volume with a cerium doped YAG crystal. A much, much brighter scintillator material than the plastic. Better performance yet will be achieved with fiber optics instead of the solid plastic light pipe.
We use John Russ's Image Processing Tool Kit (Reindeer Games Inc.) that works as Photoshop plug-ins to do this sort of thing with collagen banding. I just work on the FT image as a greyscale using Photoshop tools to select or reject the parts of interest then do a inverse filtered FT. Very easy to work with. You can remove bands or enhance bands either way.
Bob Derm Imaging Center
On Wed, 5 May 1999, Robert H. Olley wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } First, thanks to all of you who either: } } (a) told me about silver membrane filters, which I had not heard of before } (b) reminded me of Gelman, who recommended their PTFE membranes. } } As to which I use for which application, it's horses for courses! } } And now the question: one has a scanned or digitally acquired micrograph. } The background is a gently varying grey, with rather low contrast local } features (blobs, bacteria, or whatever). The task is to Fourier } transform, then filter out the lowest frequencies with a high-pass filter, } leaving one with the features on a uniform grey background. One can then } increase the contrast to make the features more prominent for printed } reproduction. Is there software (Win 95) that can do this? (preferably } inexpensive, though we could make arrangements to use plugins for Adobe } Photoshop elsewhere, if necessary). } } You can see an example of what I'm trying to do on: } } http://www.reading.ac.uk/~spsolley/fourier.htm } } +------------------------------------------------------------------------+ } | Robert H.Olley Phone: | } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } | University of Reading {University internal extension 7867 | } | Whiteknights Fax +44 (0) 118 9750203 | } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } | England URL: http://www.reading.ac.uk/~spsolley | } +------------------------------------------------------------------------+ } } }
There is a software package called EIKONA3D, developed by a company called AlphaTec Ltd., which is a 3D image processing and analysis package that provides special features and tools for working with image sequences/serial sections originated from microscopy (e.g. 3D image processing, image alignment, 3D reconstruction, 3D visualization, volume rendering, surface rendering, 3D registration,= etc.).
Further information and a free demo version of EIKONA3D are available through AlphaTec's web site: http://www.alphatecltd.com
Regards, Nikos Nikopoulos
At 02:04 =EC=EC 7/5/1999 -0500, you wrote: } } Hi everyone: } } I have been working for a long time in the field of inorganic materials.= Now } I am getting into biomaterials and my first task is to determine porosity= of } a bone, I think I need some help. I guess that if I record images of the } surface of the bone and apply sequential sectioning and image recording, I } will be able to reconstruct the 3d characteristics of the object. I wonder } if there is any software available to do the reconstruction or maybe= someone } can suggest some references where I could get started.=20 } } Thanks in advance } } } Hector Calderon }
I inquired about a classified section and was given the following response:
"There isn't a "classified" section, but you can post such a notice on the listserver. I hope that's helpful to you.
Cindy Clark MSA Administrator"
The message I want posted is as follows:
We are selling a Cambridge S100 SEM with an Energy Dispersive X-ray analysis attachment. We are looking for $5000 to $10,000 exclusive of shipping costs. Anyone interested please send e-mail to tpizzey-at-canspec.com.
We do not need to routinely capture TEM images with a digital camera, as is being discussed. It would be nice though to be able to scan negatives that are to be used for say publication or other presentation.
Is there a scanner available than can produce a high resolution image, i.e. suitable for publication?
Also, thanks to those who provided responses on basement membrane labeling.
After using our SEM's cold stage for the first time last week we have a few observations. The phenomenon which were demonstrated is with regards to cathodo-luminescence and backscatter emission, so I'll send this query to the list in two parts. I hope that someone will comment on our observations as to verify what is mysterious, and possibly explain what we are seeing.
We are examining the CL emission of quartz so as to study and possibly characterize features and textures with regards to source and history (e.g., if we were to examine a sandstone, we might be able to say the source of the qtz was metamorphic or igneous). This particular hydrothermal specimen we thought would be ideal for cold stage introduction because it had demonstrated marked areas of absolute absence of CL as well as otherwise. What we saw at low temperatures was a bit disappointing. Whereas the increase in CL intensity was very dramatic for temperatures {-100C, all of the features disappeared!!! It would be interesting to examine this phenemenon in color so as to evaluate whether the color becomes homogenous as well.
Lastly ... this particular qtz sample demonstrates a phenomenon of retaining an overprint of e-beam bombardment. That is, if we were to zoom in for a focus, a subsequent image capture would show us a small bright rectangle in the middle. This phenomenon is common, BUT it isn't demonstrated by every type of qtz. Has anyone correlated this "overprinting" with a type of qtz or possibly specimen preparation??
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
We are looking for an old Philips 201. We currently have one we are using and since parts are a little hard to come by, we would like to find another one that could be purchased for parts. If you have one you would like to sell or donate, please contact me off list.
Thanks,
Michael D. Standing Microscopy Lab Brigham Young University P.O. Box 25181 Provo, UT 84602-5181
After using our SEM's cold stage for the first time last week we have a few observations. The phenomenon which were demonstrated is with regards to ... backscatter emission .. I hope that someone will comment on our observations as to verify what is mysterious, and possibly explain what we are seeing.
We are examining the CL emission of quartz and whereas BSE imaging is of little use, we do like to document our CL imagery with an accompanying BSE image. We never expected BSE imaging at cold temperatures to be any different, so we were surprised at what was demonstrated ... which was a marked increase in the topographic contribution to the BS emission. Our BS detector is the twin solid-state type where the BS signal acquired by both halves shows atomic number contrast and the difference signal shows topography. We were using a 25mm working distance so it is difficult to believe the cold stage cooled the pole-piece mounted detector. At temperatures less than -100C we couldn't see any Z contrast at all, even for MoS inclusions in the qtz vein!!! Even at temps near zero C the polishing defects were easily seen and didn't disappear 'til ambients temps were again achieved. I am not sure the Z information actually disappears, but the topo component definitely floods the signal and is impossible to remove withour sacrificing general contrast. It would first be interesting to know if someone else has seen this with both types of BS detectors ... i.e., the scintilator type (e.g., Robinson) as well.
I've never heard of anyone speaking of this phenomenon ... does anyone have any ideas or similar observations??
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
Maybe your topographic information isn't changing (use SE at ambient and cold temps to check). But there could be a surface layer of ice which masks the atomic number contrast. You could also monitor x-ray counts at ambient and cold to see if there is a change.
: cooled the pole-piece mounted detector. At temperatures less : than -100C we couldn't see any Z contrast at all, even for MoS : inclusions in the qtz vein!!! Even at temps near zero C the : polishing defects were easily seen and didn't disappear 'til : ambients temps were again achieved. I am not sure the Z : information actually disappears, but the topo component definitely : floods the signal and is impossible to remove withour sacrificing : general contrast. It would first be interesting to know if someone : else has seen this with both types of BS detectors ... i.e., the : scintilator type (e.g., Robinson) as well. :
Carl : :
Carl Henderson Electron Microbeam Analysis Laboratory University of Michigan 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063 USA -------------------------------- Voice: (734) 936-1550 FAX: (734) 763-4690 E-mail: chender-at-umich.edu --------------------------------
Some where I seem to remember that phosphorus has enough of a radioactivity isotope to measure. Okstate built a scintillation counter that would hold a cow. You wouldn't need one nearly so bit but if phosphorus is correlated to density it would be an easy test.
While on the subject of bones I took some pictures of some rat bones being broken. It was the usual dietary calcium trial but some of the rats had been fed small amounts of vadium(sp). The rats fed vadium(sp) did not have any stronger bones than the corresponding rats in the test the only difference is while the bones broke at the same load they did not snap. The went through a very noticeable plastic deformation before separating.
I never saw any more about the research.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
-----Original Message----- } From: info {info-at-zeus.csd.auth.gr} To: microscopy-at-Sparc5.Microscopy.Com {microscopy-at-Sparc5.Microscopy.Com}
Dear Hector,
There is a software package called EIKONA3D, developed by a company called AlphaTec Ltd., which is a 3D image processing and analysis package that provides special features and tools for working with image sequences/serial sections originated from microscopy (e.g. 3D image processing, image alignment, 3D reconstruction, 3D visualization, volume rendering, surface rendering, 3D registration, etc.).
Further information and a free demo version of EIKONA3D are available through AlphaTec's web site: http://www.alphatecltd.com
Regards, Nikos Nikopoulos
At 02:04 7/5/1999 -0500, you wrote: } } Hi everyone: } } I have been working for a long time in the field of inorganic materials. Now } I am getting into biomaterials and my first task is to determine porosity of } a bone, I think I need some help. I guess that if I record images of the } surface of the bone and apply sequential sectioning and image recording, I } will be able to reconstruct the 3d characteristics of the object. I wonder } if there is any software available to do the reconstruction or maybe someone } can suggest some references where I could get started. } } Thanks in advance } } } Hector Calderon }
I would very much appreciate the complete name of the journal Boll. Zool. Please, answer me not to the list. Thanks in advance.
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA TE -(54-11)-4576-3349 e-mail micros-at-bg.fcen.uba.ar FAX (54-11)-4576-3384
I thought it was a nice touch when Nestor added the MSA plug to each of the posts and included the instructions for (un)subscribing. However, there has been a recent rash of unsubscription requests coming to the list by mistake. Apparently ones are not reading the instructions. The header plainly states that the requests are to be sent to
ListServer-at-MSA.Microscopy.Com
not to the list itself. They should have the subscription action included as the body in the form
By following these instructions, you can make the changes yourself. The only time Nestor (or the list) should have to be bothered by subscription requests is if a subscriber can no longer mail from the same address they subscribed under. In those cases, this list server (and most others) will require intervention to make sure the request is authentic. But for the majority of cases, I encourage those that wish to unsubscribe to read the header more closely. Nestor does enough for this list already.
Gatan DigitalMicrograph (DM) software can do precisely what you want to do with your images. They are all the standard features in DM. There are a large number of life science microscopists using DM. For further info, check out www.gatan.com, or send me an email.
Good luck!
Ming Pan Gatan, Inc.
} } } } And now the question: one has a scanned or digitally acquired micrograph. } } The background is a gently varying grey, with rather low contrast local } } features (blobs, bacteria, or whatever). The task is to Fourier } } transform, then filter out the lowest frequencies with a high-pass filter, } } leaving one with the features on a uniform grey background. One can then } } increase the contrast to make the features more prominent for printed } } reproduction. Is there software (Win 95) that can do this? (preferably } } inexpensive, though we could make arrangements to use plugins for Adobe } } Photoshop elsewhere, if necessary). } } } } You can see an example of what I'm trying to do on: } } } } http://www.reading.ac.uk/~spsolley/fourier.htm } } } } +------------------------------------------------------------------------+ } } | Robert H.Olley Phone: | } } | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | } } | University of Reading {University internal extension 7867 | } } | Whiteknights Fax +44 (0) 118 9750203 | } } | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | } } | England URL: http://www.reading.ac.uk/~spsolley | } } +------------------------------------------------------------------------+ } } } } } }
I am not an expert in this area but guess the procedure is as follows.
1. Use a fine metal polish to remove, by polishing, the aluminium a= nd phosphor from the old scintillator.
2. Clean with solvent the scintillator glass.
3. Mix a solution of phosphor in a solvent (?) with one or two drops=
of a plastic solution like formvar (to give strength). Ultrasonic this solution.
4. Place the scintillator in a beaker and pour onto it the phosphor solution. Cover but with a space for ventilation.
5. After some hours the solution will settle, decant off the excess and wait several days for the coating to dry.
6. Remove scintillator and if no defects are visible coat in a high vacuum, coating unit with a very thin layer of aluminium.
7. Check in microscope for efficiency.
8. If not good experiment with phosphor coating thickness and/or wit= h type of phosphor, P54 comes to mind(?)
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
Dear Gabriel and others, I have sent this to the list as some of this info is quite generally useful and not just for Gabriel's question.
Chemical abstracts service provides a list of the approved abbreviations for all the journals abstracted in "Chemical Abstracts" at http://info.cas.org/sent.html There is a list of biological journal abbreviations at: http://arachne.prl.msu.edu/journams/ and a list of journals in the ISI at: http://library.caltech.edu/admin/abbreviations/ although none of these have Boll. Zool.
Hope this helps
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
Gabriel Adriano Rosa wrote: } I would very much appreciate the complete name of the journal Boll. Zool. } Please, answer me not to the list. } Thanks in advance.
I am not in the biology sector, but I guess it should be Bull. Zool. - there is at least a journal named Raffles Bulletin of Zoology.
Joergen.
J. B. Bilde-Soerensen Senior Research Scientist, Ph. D. Materials Research Department Risoe National Laboratory DK-4000 Roskilde Denmark
I am trying to visualize by negative staining the attachment of phages to bacteria. However the bacteria shrink under the EM vacum, leaving an artefactual space between the negative stain and the cell wall which do not allow to take good pictures of the attached phages.
Here is the protocol I am using: I am using 2% uranyl acetate as the negative stain The cells are adsorbed to carbon coated formvar grids for 1 minute Excess is drained with filter paper The grids are floated in the negative stain for 1 minute Excess is drained with filter paper The grid goes into the TEM
I have tryed a few modifications such as fixing the material and mixing the stain with the bacteria suspension, but it did not work.
can anyone send further suggestions???
Sincerelly
Dr. A.P. Alves de Matos EM Unit, Pathology Department Curry Cabral Hospital Lisbon
Does anyone know the formula/ chemical name for an etchant called glyceregia? It is used to show carbonitrides in stainless steels for optical microscopy and I'ld like to know how to prepare some.
Im forwarding this message to the list. Hopefully someone in the states will be able to help.
Please email him direct at pwilsonarzu-at-Hotmail.com
} Dear Sir: } } I would like some information on buying a used electron microscope in the } United States. } } I would really appreciate any information you may have available. } } My e-mail: pwilsonarzu-at-Hotmail.com } } Thank you in advance, } } Philip E. Wilson
---------------------- Kevin Mackenzie Electron Microscope unit Dept Zoology University of Aberdeen Tillydrone avenue Aberdeen AB24 2TZ ----------------- Tel 01224 272847 Fax 01224 272396 email k.s.mackenzie-at-abdn.ac.uk
Hi everyone, I have heard of radiation damage to the specimens, and I'm curious how much beam current and current densities are there on the specimen in HREM imaging and nano-diffraction. Your replies are highly appreciated.
A search of the library here turned up "Bollettino di zoologia" published as vols. 1-62, 1930-1995, and since continued as the Italian Journal of Zoology.
Best regards,
Dan Luchtel, Ph.D. Professor University of Washington Environmental Health Box 357234 Seattle, WA 98195-7234
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Good day all, Does anyone have a schematic for a Coolwell chiller? Model # SE-075W C2?? I need some help to find out what the electronics are suppose to be? not what they are. Any help would be nice, since what I have heard they where bought out, and the holding company only wanted the name and management, so no tech help. Sincerly,
************************************************ Robert J. Derby New Mexico Institute of Technology Socorro, N.M. Phone - 505-835-5866 E-mail - rjderby-at-excite.com derby-at-nmt.edu ************************************************
_______________________________________________________ Get your free, private email at http://mail.excite.com/
} Some where I seem to remember that phosphorus has enough of a } radioactivity isotope to measure. Okstate built a scintillation counter } that would hold a cow. You wouldn't need one nearly so bit but if } phosphorus is correlated to density it would be an easy test. } Dear Gordon, Natural phosphorus is 100% P-31. The two radioactive isotopes are P-32 & P-33 with half-lives of 14 & 25 days, respectively. They both emit relatively high-energy beta-. Even so, the beta's would not make it to a scintillation counter if they originated in cow bones (assuming that the rest of the cow was still attached :-)). You are probably thinking of potassium, which has a significant amount of K-40. It is this isotope which gives a measurably larger dose of radiation from being inside a concrete building, as opposed to a wood building. Yours, Bill Tivol
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} I am trying to visualize by negative staining the attachment of phages to } bacteria. } However the bacteria shrink under the EM vacum, leaving an artefactual } space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain } The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper } The grids are floated in the negative stain for 1 minute } Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions??? } Dear A.P., I am most definitely not an expert on this. The protocol you're using is the same as I use in my work. Since the problem is with shrink- age, perhaps placing a few ul of bacterial suspension on the grid, air drying and placing it in a vacuum, then applying the UAc will prevent the problem. Of course, such treatment could induce artifacts--it is a pretty severe process--but it might still tell you what you want to know. Freeze-drying could be another way to go, but this can also pro- duce artifacts; I've seen the results when this was done to kidney, and it's not pretty. Good luck. Yours, Bill Tivol
Hello! I need a heater to keep mammalian cells warm while viewing them with a light microscope. I know there are various types avaiable but I'm not sure where to find them. Can anyone recommend a company that sells this sort of thing? Cheers, Jennifer
-- Jennifer Shuler, Ph.D. Director of Imaging Facility Adjunct Assistant Professor Biology Department, Box 7325 Wake Forest University Winston-Salem, NC 27109
Adam wrote: } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello, } } Does anyone know the formula/ chemical name for an etchant called } glyceregia? It is used to show carbonitrides in stainless steels for } optical microscopy and I'ld like to know how to prepare some. } } Thank you, } } Adam Scott } } ascott1-at-engfac.uct.ac.za
Adam,
Please go to the http://www.kaker.com/mvd/vendors.html, there is a link to my free etchant's database. Bellow is search result:
Material: Wrought stainless steels (Fe) Type: Microetchant Method: Chemical etching Etchant (Electrolyte): 10 ml HNO3, 20 to 50 ml HCl, 30 ml glycerol (Glyceregia). Procedure: Mix HCl and glycerol thoroughly before adding HNO3. Discard before solution attains a dark orangr color. Immerse or swab specimen for a few seconds to a few minutes. Higher percentage of HCl minimizes pitting. Remarks: General structure. Reference: Metallography, Structures and Phase Diagrams, Metals Handbook, 8th Edition, Vol. 8, ASM (American Society for Metals), Metals Park, Ohio 44079, USA, 1973, p. 98.
Henrik
-- Henrik Kaker, Ph.D. SEM-EDS Laboratory Metal Ravne, Koroska cesta 14 Ravne, Slovenia, Phone: +386 602 21 131 Fax: +386 602 20 436 Mailto:Henrik.Kaker-at-guest.arnes.si http://www2.arnes.si/guest/sgszmera1/index.html
} -----Original Message----- } } Hello, } } Does anyone know the formula/ chemical name for an etchant called } glyceregia? It is used to show carbonitrides in stainless steels for } optical microscopy and I'ld like to know how to prepare some. } } Thank you, } } Adam Scott } } ascott1-at-engfac.uct.ac.za ------------------------------------------------------
Adam,
According to the ASM Metals Handbook, 8th Edition, Volume 8, page 98
"Glyceregia: 10ml HNO3, 20 to 50 ml HCL, 30 ml glycerol."
"Procedure: Mix HCL and glycerol thoroughly before adding HN03. Discard before solution attains a dark orange color. Immerse or swab specimen for a few seconds to a few minutes. Higher percentage of HCL minimizes pitting"
Hope this helps.
Neither TIMET nor William Giles assume any responsibility for the above quoted formula.
This is a follow-up on my query about fixing isolated mitochondria for TEM. First, thanks to all who responded. A summary of replies is below. We solved our immediate problem by fixing in 4% glutaraldehyde in 0.1M sodium cacodylate, r.t. for ca 30 min, and then into the fridge until the next morning. I followed with a fairly routine postfixation, dehydration, and embedding. We got good enough results to answer the first questions. However, there will be many more!
I would like to mine the collective expertise further. For the next step the client would like to localize calcium and, if possible, do some crude quantitation. From my background in neurophys I know this is tricky. Do any of you have a current favorite localization technique? In the (far distant) past I have used K-pyroantimonate to precipitate Ca with mixed results, and I can't find that particular recipe. New ideas would be enthusiastically received.
Soon I hope to have a better instrument to help in this and many other quests. We just installed a LEO (Zeiss) 912 AB TEM with integrated omega energy filter. What a beautiful instrument! What potential! How exciting! I wish I knew how to use it... I've only had it a couple of weeks and, while I can make the basic instrument work well, I still haven't had my "applications training" to learn how to use the energy filter for ESI, EELS, and all the really useful and fun stuff. I've tried to bull my way through the manuals and run on intuition, but that approach isn't working...
However, I am confident it will be useful for this kind of problem. I would be interested in striking up an e-mail penpal relationship with anyone else with this instrument.
We do not yet have a cryostage for it, and so looking at ultrarapidly cryofixed, unprecipitated ions is not yet a possibility.
Secondly, I would like to hear from anyone who might have an explanation and/or citations for the appearance of these isolated mitochondria that have been subjected to high Ca concentrations. We are seeing cup-shaped mitochondria (which I see in my inverts all the time), but the outer membrane on the side of the concavity is NOT concave, and is swollen outward. This wouldn't bother us so much, except it nags at me that there is something asymetrical about the mitochondria membranes that causes the inner and outer membranes to stay attached on one side and not on the other. Plus, sometimes the cristae are swollen and sometimes not. I would appreciate being pointed to appropriate literature, if any. My searches have not been very fruitful at this point. Control mitochondria processed at the same time do not exhibit this morphology.
Here is the summary of responses I received a few weeks ago:
Have you tried/considered fixing them in the last buffer they see before fixation? I always worry about isolation protocols....the mitos may look blah because of something they've been in during the isolation protocol, not because of your solutions. Now I try to at least match the final solution in the prep. As an example - organelles that have been purified through sucrose gradients are going to look like crap if they are taken from their sucrose and fixed in 0.1M any buffer! (Been there, done that...). And mitos are so fussy, anyway.......Investigators can be reticent about their preps. I'm sure you've had the, "Oh, didn't I tell you? These fractions are in 5M Tris" experience. Homicide doesn't seem to be a viable response :) ********************************************************** Yes I did, thanks. They looked really terrible! ********************************************************** A long time ago back in the mists of time I used to fix isolated cell fractions, mitochondrial, lysosomal, nuclear, gap junctions, etc.
We used a double fixative of cold Glutaraldehyde and Osmium Tetroxide mixed.
Reference as follows:-
Ultrastructure of Human Leukocytes after simultaneous fixation with Ultrastructure of Human Leukocytes after simultaneous fixation with Glutaraldehyde and Osmium Tetroxide and "post-fixation" in Uranyl Acetate. Hirsch, J.G. & Fedorko, M.E. (1968) J.Cell. Biol. 38, 615.
This excellent for cell suspensions and fractions. The fixative is mixed on ice and used immediately after mixing. Fixation also on ice. Fixation time for a small pellet about 30 minutes.
I used this method a lot at one time and can thoroughly recommend it. But, only for suspensions and cell pellets. Not for solid tissue.
Membranes are well defined, swelling/shrinkage artefacts are not a problem. ******************************************************************* We have had good luck with high-pressure freezing. We have examined unstained, frozen-hydrated mitos on the 400 kV instrument here. Although this procedure requires instruments not available at all facilities, I expect to get minimal artefact. ******************************************************************
Mahalo to you all- 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* ****************************************************************************
You better check on the polishing step with someone who knows for sure. If there is a conductive coating on the glass under the phosphor (and I'm sure there is), such as tin oxide or indium tin oxide, you'll probably take that off too. The thin aluminum on top of the phosphor protects the phosphor and is not used as the applied voltage contact. Hard rubbing without the abrasive will not take the coating off the glass. A product like Soft Scrub might do the trick for you, but again, I would double check before I would risk it.
You have to be careful in Steve's step 5. If you cause the water solution to move too much, you can get a wavy coating. One trick that I used when making transparent screens, is to run a small diameter tube into the container that the plate being coated was place down to the bottom of the bottom. I would then tape the tube to the outside and attach a syringe to the bottom. The syringe was below the bottom of the container with the tube and syringe full. When it was time to decant the liquid, I pulled the plunger and started a siphon into a beaker. It took a while because the tubing was so small, but it worked fine.
Another addition that you can put in the solution is potassium silicate. (Sometimes this is called liquid glass.) It helps to bond the phosphor to the glass. Sorry, but I can't remember how much to put in. It was only a few percent.
-Scott
---------- } From: Steve Chapman To: de Lillo Enrico; American -----------------------------------------------------------------------.
I am not an expert in this area but guess the procedure is as follows.
1. Use a fine metal polish to remove, by polishing, the aluminium and phosphor from the old scintillator.
2. Clean with solvent the scintillator glass.
3. Mix a solution of phosphor in a solvent (?) with one or two drops of a plastic solution like formvar (to give strength). Ultrasonic this solution.
4. Place the scintillator in a beaker and pour onto it the phosphor solution. Cover but with a space for ventilation.
5. After some hours the solution will settle, decant off the excess and wait several days for the coating to dry.
6. Remove scintillator and if no defects are visible coat in a high vacuum, coating unit with a very thin layer of aluminium.
7. Check in microscope for efficiency.
8. If not good experiment with phosphor coating thickness and/or with type of phosphor, P54 comes to mind(?)
Steve Chapman
Senior Consultant E.M. Protrain, 16 Hedgerley, Chinnor, Oxford OX9 4TN, England. Tel & Fax 44 (0)1844 353161 Web Site - http://ourworld.compuserve.com/homepages/protrain For Consultancy and Courses in Electron Microscopy World Wide
First, try the company from which you purchased your microscope. Secondly, BioOptechs has great stages for live cell work. Finally, there are several manufacturers for warming stages. Visit either www.mwrn.com or the microscopy society site (www.msa.microscopy.com) for vendors. Email me if you have trouble finding contact information.
Best regards Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 02:34 PM 5/11/99 -0400, Jennifer Waters wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} I am trying to visualize by negative staining the attachment of phages to } bacteria. } However the bacteria shrink under the EM vacum, leaving an artefactual } space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain } The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper } The grids are floated in the negative stain for 1 minute } Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions??? }
You can freeze-dry the negative stains. Look for the papers of N.V. Nermut e.g. in the Proc 5th European Congress on Electron Microscopy 1972 page 236. ***************************************************** Mel Dickson, Deputy Director. Electron Microscope Unit, University of New South Wales. Sydney NSW 2052 Australia
Has anyone had success in staining wax in a rubber matrix? Either optical or electron contrast methods would be of interest, particularly those for SEM. Thanks for your thoughts.
Hi all, Boll.Zool. stands for Bollettino di Zoologia, now known as "The Italian Journal of Zoology". Here's their website: http://www.uniroma1.it/bau/uzi/itjz.htm
Glyceregia formula =3D 10 ml HNO3 + 20 to 50 ml HCl + 30 ml glycerine (ref.: ASM Metal Handbook, Desk Edition, page 35.35)
Marc Montreuil Rolls-Royce Canada
Adam a =E9crit:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=
} To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------= =2E } } Hello, } } Does anyone know the formula/ chemical name for an etchant called } glyceregia? It is used to show carbonitrides in stainless steels for } optical microscopy and I'ld like to know how to prepare some. } } Thank you, } } Adam Scott } } ascott1-at-engfac.uct.ac.za
-----Original Message----- } From: William Tivol {tivol-at-wadsworth.org}
} Dear Gordon, } Natural phosphorus is 100% P-31. The two radioactive isotopes } are P-32 & P-33 with half-lives of 14 & 25 days, respectively. They both } emit relatively high-energy beta-. Even so, the beta's would not make } it to a scintillation counter if they originated in cow bones (assuming } that the rest of the cow was still attached :-)). } You are probably thinking of potassium, which has a significant } amount of K-40. It is this isotope which gives a measurably larger dose } of radiation from being inside a concrete building, as opposed to a wood } building.
Bill,
I always did have trouble mixing up P & K when it was listed in any format but N-P-K in fertilizer.
If we wanted to measure P-32 or 33 we would have to drop the cow in the whole cow grinder. They have one that will grind up a cow hide, hair, guts and all for finding out what actually makes up a cow. It wouldn't be very useful for estimating bone destiny but the K-40 might if it is correlated to bone density. It would probably give skewed results for a lot of the world due to Chernoble and nuclear testing.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
Bacteria are a bit thick for negative staining. Changing the stain concentration helps a little. The rule is that thick specimens appear better with a lower concentration of stain, say 2%. Thin specimens are better with a lower concentration, say 0.5%. You may get better appearance trying different staining solutions, PTA or Ammonium molybdate are good candidates. Don't grow cultures on a shaker, many flagella drop off and even fimbria may suffer.
This is a difficult subject for negative staining because you require both, bacteria and flagella in one picture. Much prettier images can be obtained with shadow casting. 1 Apply bacteria solution to the face of substrated grid. (apply and blot lightly several times to obtain better distribution) Drying would leave too much debris and salts, so use a volatile buffer to wash and maintain molarity. 2 Apply and blot bacteria coated grid repeatedly to a 0.1 M solution of ammonium acetate. 3 After air drying, angle (or rotary) shadow) grid in a vacuum evaporator using fine grain evaporating material. (simultaneous C/Pt or if available high resolution Cr coater as used for FESEM)
Another alternative: If the equipment is available would be FESEM of bacteria and flagella and this could be supplemented with an image of negatively stained flagella only, taken in TEM.
Its a little challenging project, but some nice images could result. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
} I am trying to visualize by negative staining the attachment of phages to } bacteria. } However the bacteria shrink under the EM vacum, leaving an artefactual } space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain } The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper } The grids are floated in the negative stain for 1 minute } Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions???
you can try to adsorb your bacteria/phage mixture onto carbon film, freshly prepared onto cleaved mica. Just cut out a small piece of the carbon film, a little bit smaller than your grid diameter. Put it in the sample solution under a 45=B0 angle and the carbon film will float from the mica. Take care that not all the carbon film floats of the mica! Let the samples adsorb for about 30 sec or 1 min depending on the optical density of your sample. Take the piece of mica back from the solution; the carbon film will reattach to the mica. Rinse in TE-buffer (Tris-EDTA buffer, 20 mM Tris, 1 mM EDTA,. pH 7.0)several times. Subsequentely, float the entire carbon film on 4% aqueous uranyl acetate, leave it for 10 sec and then pick up the carbon film with your grid. Soak access staining solution to such an extent that the surface of the grids is just wet; than immediately dry the grid under a lamp. You can also try to vary the amount of staining solution residing on the grid, i.e. to perform a "deep" stain and a "shallow" stain. Good luck. Manfred
At 12:50 11.05.99 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America=20 the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions??? } } } Sincerelly } } Dr. A.P. Alves de Matos } EM Unit, Pathology Department } Curry Cabral Hospital } Lisbon } } } } }
Although the formulation for GLYCEREGIA is not the same as that for for making Nitroglycerine, nevertheless mixtures of alcohols (and other organics) with nitric acid need to be handled with care.
In the present case, I would not expect an explosion, but as in Henrik Kaker's reply, these mixtures can turn a dark orange colour. This is a warning that there may be a runaway reaction impending - generally the stuff boils over with copious quantities of brown nitric oxide gas being given off. The quantity of HCl in the mixture should prevent that happening too suddenly, but DO work in a fume cupboard and DON'T leave such a mixture around unattended.
+------------------------------------------------------------------------+ | Robert H.Olley Phone: | | J.J.Thomson Physical Laboratory {direct line +44 (0) 118 9318572 | | University of Reading {University internal extension 7867 | | Whiteknights Fax +44 (0) 118 9750203 | | Reading RG6 6AF Email: R.H.Olley-at-reading.ac.uk | | England URL: http://www.reading.ac.uk/~spsolley | +------------------------------------------------------------------------+
I obtained quite good results on E.coli with T4 phage by fixing in 2.5% glutaraldehyde, washing and then staining with about 1% sodium silicotungstate. There were no gaps and staining was good enough to visualise bacteria and fine structure of phage. I used sodium silicotungstate because it seems OK for both bacteria and viruses and seems better behaved than PTA or molybdate and I usually find that uranyl acetate can be very unpredictable in terms of quality and consistency and doesn't keep well.The reason I fixed wasn't to improve preservation - it was just so I could do a time-scale for the infection cycle to produce sections and negative stains
It was over 10 years ago and it's just my opinion, anyway.
Malcolm Haswell Electron Microscopy School of Sciences Fleming Building University of Sunderland SUNDERLAND SR1 3SD Tyne and Wear UK
Tel (0191) 515 2872 e-mail: malcolm.haswell-at-sunderland.ac.uk
---------- } From: jim-at-proscitech.com.au To: A.P. Alves de Matos Cc: 'microscopy-at-sparc5.microscopy.c
teria are a bit thick for negative staining. Changing the stain concentration helps a little. The rule is that thick specimens appear better with a lower concentration of stain, say 2%. Thin specimens are better with a lower concentration, say 0.5%. You may get better appearance trying different staining solutions, PTA or Ammonium molybdate are good candidates. Don't grow cultures on a shaker, many flagella drop off and even fimbria may suffer.
This is a difficult subject for negative staining because you require both, bacteria and flagella in one picture. Much prettier images can be obtained with shadow casting. 1 Apply bacteria solution to the face of substrated grid. (apply and blot lightly several times to obtain better distribution) Drying would leave too much debris and salts, so use a volatile buffer to wash and maintain molarity. 2 Apply and blot bacteria coated grid repeatedly to a 0.1 M solution of ammonium acetate. 3 After air drying, angle (or rotary) shadow) grid in a vacuum evaporator using fine grain evaporating material. (simultaneous C/Pt or if available high resolution Cr coater as used for FESEM)
Another alternative: If the equipment is available would be FESEM of bacteria and flagella and this could be supplemented with an image of negatively stained flagella only, taken in TEM.
Its a little challenging project, but some nice images could result. Cheers Jim Darley ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes ********************** www.proscitech.com.au *****
} I am trying to visualize by negative staining the attachment of phages to bacteria. However the bacteria shrink under the EM } vacum, leaving an artefactual space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper The grids are floated in the negative stain for 1 minute Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions???
I have had good results by mixing a drop of1% phosphotungsic acid (PTA) with an equal amount of my liquid bacterial culture on a glass slide, and drying about 0.5 mucroliters of this mixture on a formvar coated grid. You can eliminate distracting salt crystals by first gently centrifuging the bacteria (4 minutes at 5K rpm in an epindorf microcentrifuge) and resuspending the pellet in distilled water. You may not get away with this and still have phages adhering to the cells, but you could try before or after adding the phages. Good luck.
I have always made my lead citrate according to Reynold's original protocol starting with lead nitrate and sodium citrate. Somebody borrowed my bottle of lead nitrate and never returned it but I have a fresh bottle of granular lead citrate. I know there is a modification of Reynold's that starts with lead citrate. Does anyone have the formula and/or opinion on whether it is any different than the original. Thanks, Tom
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
The protocol I have used in the past is to hold the grid in forceps, apply about 10-20 ul of bacterial suspension for 1 min, and then draw off with filter paper. I then apply 20 ul of 2% aqueous phosphotungstic acid, leave on about 10 sec. and draw off and air-dry. You may have to fool around with the dilution of the bacterial suspension, so it's not too thick. Good luck.
Mary McKee
On Tue, 11 May 1999, A.P. Alves de Matos wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear collegues } } I am trying to visualize by negative staining the attachment of phages to } bacteria. } However the bacteria shrink under the EM vacum, leaving an artefactual } space between the negative stain and the cell wall which do not allow to } take good pictures of the attached phages. } } Here is the protocol I am using: } I am using 2% uranyl acetate as the negative stain } The cells are adsorbed to carbon coated formvar grids for 1 minute } Excess is drained with filter paper } The grids are floated in the negative stain for 1 minute } Excess is drained with filter paper } The grid goes into the TEM } } I have tryed a few modifications such as fixing the material and mixing the } stain with the bacteria suspension, but it did not work. } } can anyone send further suggestions??? } } } Sincerelly } } Dr. A.P. Alves de Matos } EM Unit, Pathology Department } Curry Cabral Hospital } Lisbon } } } }
HREM data is fairly easy to work out. We would generally use a 2 second exposure at 300k to 500k times mag. For our film (at 400kV) 20pA/cm2 gives the correct exposure. Ignoring the beam absorbed by the specimen, which should be thin for HREM, this works out at around 2x10^7 to 5x10^7 A/m2 on the specimen (assuming my maths is OK). This should be a reasonable estimate of the current density on the specimen. The beam area would typically be of the order of 0.3um to 1um diameter (approx. 10^-12 to 10^-13 m2).
Nano diffraction is much more variable - it depends on the type of electron gun, probe size, probe defocus, energy spread, etc. However, as a guide we can get a current of 1nA into a spot of 1nm for a FEG which is claimed to be 100 times brighter than a LaB6.
I hope this helps.
Regards, Ron
On Tue, 11 May 1999 09:35:09 -0500 (CDT) WENTAO QIN {s987041-at-jinx.umsl.edu} wrote: } } Hi everyone, } I have heard of radiation damage to the specimens, and I'm curious } how much beam current and current densities are there on the specimen in } HREM imaging and nano-diffraction. Your replies are highly appreciated. } } Wentao } }
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
Call Frank Haze, 800-367-5665. He can furnish you with whatever information is available for your Coolwell chiller. He also has coolant fluid still available. Good luck!
I have been using an HP Scanjet 4c which is about 4 years old and has a resolution of 300dpi. I find that this gives excellent results when printed on my Codonics 1600 dye sub printer. In fact I have found that scanning in at 150 to 200 dpi is sufficent in most cases and results in files of 1/2 the size of a 300dpi file. I may be mistaken but I believe the resolution of most prints in journals is 75 to 125 dpi, so if you scan in at 200 and send them an electronic version, that should suffice.
If you need photographic quality prints, there are numerous printers (ie Epson, HP, etc) that rivel photographs. My only complaint is the paper they use is far from being similar to actual photographic paper.
I am working on making AFM topography and phase images of a series of polymer samples which are made up of varying amounts of polycarbonate, polypropylene and mostly (} 80%) ABS. I have some nice images already (soon to be on our web site) but want to improve the quality. I also want to look at the bulk regions by using cryo-microtomy to create a nice flat face for imaging. Can someone share their experiences with these types of samples and any tips on the microtomy technique (best temperatures, etc.)? I also plan to use plasma etching to see if that will clean up the surface.
Thanks in advance.
Robert J. Plano Staff Analyst, SPM Services Surface Science Labs (650)962-8767, ext. 742
Dear Tom, The formula you are looking for is that of Venable, J. M., and Coggeshall, R. (1965). A simplified lead citrate stain for use in electron microscopy. J. Cell Biol. 25, 407. Add 0.01-0.04 g of lead citrate and 0.1 ml of 10 N NaOH to 10 ml distilled water to a screw-topped vial. Use distilled water that has been freshly boiled for at least 5 min. to ensure solutions are CO2-free. The mixture should be sonicated or vigorously shaken until the lead citrate is dissolved. Centrifuge before use. Use stain immediately. Usual staining time is 1-5 min. My experience is that this formula is much more intense than Reynolds (1963) lead citrate formula. This is helpful for getting more contrast from Spurrs resin sections for which I use 0.035g of lead citrate and stain for 5 -7 min. Word of caution: this stain is very sensitive to CO2. Thus to avoid precipitation make fresh each time of use and keep staining times short, and rinse well in CO2-free distilled water.
} I know there is a modification of Reynold's that starts with } lead citrate. Does anyone have the formula and/or opinion on whether it is } any different than the original.
I think the one you are looking for is the formulation described by Venable and Coggeshall (1965) (J.Cell.Biol. 25, 407-408). I haven't checked the original reference but in our notes the instructions are: weigh out 0.02g lead citrate, add 10ml distilled water followed by 0.1ml 10M NaOH. Shake vigorously until the solution is clear. Allow to stand for at least 30 mins and do not agitate the bottle.
It is also important not to use any of the staining solution from close to the bottom of the bottle.
We used this stain for a while many years ago but reverted to Reynolds which we have found to be less prone to contamination.
I seem to remember that there is another formulation using lead citrate as the starting material, by Sato, I think, but I cannot find it right now!
Regards
Robin
} } 3 Tucker Hall } Division of Biological Sciences } University of Missouri } Columbia, MO 65211-7400 } (573)-882-4712 (voice) } (573)-882-0123 (fax) }
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za (or eurc-at-giraffe.ru.ac.za) tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
This has possibly been covered so I'll apologise in advance.
Does anyone out there know of a method to accurately determine the thickness of carbon films used on TEM grids? EELS is a possibility but I was looking for a more direct measurement eg using an SPM.
Thanks very much.
Colin Veitch
Instrumentation Scientist Electron Microscopy Textile and Material Technology Group CSIRO Textile and Fibre Technology PO Box 21, BELMONT, Vic. 3216. Australia.
Hello Malcolm and other involved in this discussion
} I obtained quite good results on E.coli with T4 phage
It will always be difficult to optimize negative stain for both viruses and bacteria because they are so different in size and density. For those who would like to see what I mean have a look at some results from one of our recent Microbiology undergrad practicals (http://www.ru.ac.za/emu/im4301.htm) - low res pictures, I'm afraid (can't afford a slow scan CCD camera!) but they do illustrate how the staining conditions vary for the different structures involved.
Regards
Robin
Robin H Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa R.Cross-at-ru.ac.za (or eurc-at-giraffe.ru.ac.za) tel: +27 46 603 8168 - fax: +27 46 622 4377 http://www.ru.ac.za/emu/index.htm
I have been looking at some digital cameras for optical microscopy that "integrate". I would have called it "frame averaging" or "pixel averaging", but it appears to be the same image pre-processing technique. Specifically, I saw an "Optronics" camera and it seemed to be able to process low light levels as low as many dark field situations without the memory affects seen on low lux type cameras. Has anyone had any experience with this particular trade name? I am looking for service and dependability technical issues. Also any similar cameras? Please respond to my email. I don't want to flood the server with this issue,especially if any comments are slightly critical or complimentary and appear commercially biased. I don't mind commercial responses. I believe they are a real necessary technical component of this and all issues. Vendors are some of the best and most motivated technical resources we have available to us.(..but that's another subject...)
(I have no commercially beneficial interest in any cameras except the benefits of my laboratory applications, that is I do not sell them. "I am not spam."....did I cover this?)
jeffrey/ 'JD'
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
I have been looking at some digital cameras for optical microscopy that "integrate". I would have called it "frame averaging" or "pixel averaging", but it appears to be the same image pre-processing technique. Specifically, I saw an "Optronics" camera and it seemed to be able to process low light levels as low as many dark field situations without the memory affects seen on low lux type cameras. Has anyone had any experience with this particular trade name? I am looking for service and dependability technical issues. Also any similar cameras? Please respond to my email. I don't want to flood the server with this issue,especially if any comments are slightly critical or complimentary and appear commercially biased. I don't mind commercial responses. I believe they are a real necessary technical component of this and all issues. Vendors are some of the best and most motivated technical resources we have available to us.(..but that's another subject...)
(I have no commercially beneficial interest in any cameras except the benefits of my laboratory applications, that is I do not sell them. "I am not spam."....did I cover this?)
jeffrey/ 'JD' Email: WA5EKH-at-juno.com
___________________________________________________________________ You don't need to buy Internet access to use free Internet e-mail. Get completely free e-mail from Juno at http://www.juno.com/getjuno.html or call Juno at (800) 654-JUNO [654-5866]
Dear Microcopists: I am doing HREM image simulation to compare my HREM image obtained by Philips EM 300 FEG. The parameters needed for calculation are: spherical aberration coeffienct (Cs), Gaussin defocus ("DELTA" for the Defocus spread ), beam convergent semiangle. I appreciate it if any one can give me the numbers of these three parameters for EM 300 FEG microscope. Thank you in advance.
You really need to measure these yourself as they do vary from 'scope to 'scope. I would first get hold of an amorphous Ge sample and do a tilted series of high resolution images for equal and opposite beam tilts (make sure the tilts are calibrated). If you take four tilts +/- X and +/- Y directions you can extract C_s. Using a higher number of tilts you can extract the third order aberration coefficient of your objective lens as well. Here are some references that may help you:
1: Measurement of the spherical aberration coefficient of TEMs by beam tilt induced image displacement: A.J.Koster & A.F. deJong: Ultramicroscopy 38 (1991) pp235-240
2:Improved methods for the determination of the spherical aberration coefficient in HREM from micrographs of an amorphous object: W.M.J.Coene & T.J.J.Denteneer: Ultramicroscopy 38 (1991) pp 225-233
3:Three fold astigmatism in HREM: O Krivanek Ultramicroscopy 55 (1994) pp419-433
4:A spherical aberration corrected 200 keV TEM M Haider, H Rose, S Uhlemann, E Scwann, B Kabius & K Urban Ultramicroscopy 75 (1998) pp53-60
Number 4 is definitely worth a look since it has some nice pictures to demonstrate.
-- ***************************************** Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
Tom, Oops. I gave you the wrong amount for lead. I should proof my own messages more carefully. It's 0.04 gms of lead citrate , not .4 like I previously said. Also if you place sodium hydroxide pellets around the perimeter of the staining dish it will absord the CO2.
I only dream about 300 keV FEG TEMs, so I don't have the numbers right handy. If you don't get them from the list, you might want to get a hold of J. C.H. Spence's book (Experimental High Resolution Electron Microscopy( Amazon says it's out of print)). As I recall there are step by step procedures for determining at least some of these values. A test sample like a holey gird that has seen a few seconds in an Au sputter-coater will help too.
cheers, John MSU
} Dear Microcopists: } I am doing HREM image simulation to compare my HREM image obtained by } Philips EM 300 FEG. The parameters needed for calculation are: } spherical aberration coeffienct (Cs), Gaussin defocus ("DELTA" for the } Defocus spread ), beam convergent semiangle. } I appreciate it if any one can give me the numbers of these three parameters } for EM 300 FEG microscope. Thank you in advance. } } DAI Jiyan } IMRE
I would like to thank those who send any information about the complete name of the journal Boll. Zool. I would also like to tell that it was not a missprint and that the journal=B4s name is Bolletino di Zoologia now known as The Italian Journal o= f Zoology (Ital. J. Zool.) since 1996. Bye.
Gabriel Adriano Rosa Area Microscopia Electronica, Depto. Cs. Biologicas Fac. de Ciencias Exactas y Naturales, Universidad de Buenos Aires Ciudad Universitaria, 4 piso, Pab. II, CP 1428, Buenos Aires, ARGENTINA TE -(54-11)-4576-3349 e-mail micros-at-bg.fcen.uba.ar FAX (54-11)-4576-3384
Hi All, We have a Zeiss TEM. Normally when the filament is first switched on with both current and heating knobs are turned to mininimum the voltage indicator shoots up to 2V then immeadiatly starts going down to 1.3-1.4V within no more than 2 minutes, then we start raising the voltage by turning the heating knob until the voltage meter reads between 1.5-1.6V (the recomended voltage stated in the manual). Recently though we have been noticing that the voltage shoots up to 2.2V and stays there for several minutes then starts coming down very slowly and settles at more than 1.6V!!... Needless to say that we have also been burning too many filaments too quickly!! The filament lives have been no more than 10-15hours. We would appreciate any hints or suggestions on why would this happen. We are suspecting an electric problem in the filament control circuitry. Thank you.
NORAN Instruments, Middleton, WI, is a manufacturer of microscopy and microanalysis instrumentation and detectors. Check out our Web site at {www.noran.com} to see what kinds of products we are involved in.
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NORAN is actively seeking qualified software engineers or senior software engineers to support existing products (which are both UNIX and NT-based), and to assume a role in new product development. Mostly C and C++ programming at present. Familiarity with scientific programming in the areas of microscopy, imaging, microanalysis, or spectroscopy would be a definite plus.
NORAN is an Equal Opportunity Employer.
Benefits and compensation are excellent!
Middleton is a beautiful suburb of Madison, and considered by many to be one of the choicest communities in this area. Madison itself has been named as the best place to live in America. Situated between two large lakes offering many recreational opportunities, Madison is replete with many nice parks and educational institutions (including the main University of Wisconsin campus), among other attractions, which contribute to an excellent overall quality of life.
If you or someone you know are qualified for and interested in any of these positions, please contact: Human Resources NORAN Instruments 2551 West Beltline Highway Middleton, WI 53562-2697 Phone: (608)831-6511 FAX: (608)836-7224 Or send your resume to me personally and I will gladly forward it to HR for you.
-------------------------------------------------- Timothy G. Moeller | NORAN Instruments Inc., Sr. Software Engineer | a ThermoSpectra company {tmoeller-at-noran.com} | {www.noran.com} -------------------------------------------------- DISCLAIMER: The statements and opinions expressed here are my own, and may not represent those of my employer, NORAN, nor of its parent, ThermoSpectra.
We routinely use lead citrate as described in a method by Venable and Coggeshall (1965) in J.Cell Biol.,25: 407 . It is quick to make up and always clean as long as you only use it on the day of making. We keep pre weighed portions of lead citrate ( 0.1 to 0.4 gm) in 10 ml specimen tubes When we need the stain we add 1 ml of N1 sodium hydroxide , wait a few minutes for the lead citrate to dissolve and add 9 ml of distilled water. Use only carbonate free NaOH and fresh distilled water.
Regards
John Manston John Manston Electron Microscope Unit Division of Biomedical Sciences Queen Mary and Westfield College University of London Mile End Road London E1 4NS Tel +171 982 6961 Fax +181 983 0613
We have a Aeiss 10A for sale. Duke University Equipment Appraisal Office and appraised it at $10K. Address at bottom. S Miller
On Thu, 15 Apr 1999, Peter Jordan wrote:
} Date: Thu, 15 Apr 1999 22:26:29 -0700 } From: Peter Jordan {emsi-at-pe.net} } To: EM Listserver {Microscopy-at-sparc5.microscopy.com} } Subject: Used Zeiss 10C } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All: } I am still looking to buy a used Zeiss 10 TEM. There must be one out } there for sale. Please let me know. Thanks. } Peter Jordan, EMSI } } }
Sara E. Miller, Ph. D. P. O. Box 3020 Duke University Medical Center Durham, NC 27710 Ph: 919 684-3452 FAX: 919 684-8735
since you requested information through email, we will send you information about our high resolution digital light microscope cameras that way. If anybody else is interested in getting this information, please give us a call or contact us otherwise.
A technical note: Integrating is not necessarily the same as frame or pixel averaging. Frame or Pixel averaging means, that the camera acquires several frames and they are averaged on the frame grabber or the computer. Integration means, that the camera integrates the light on the chip by allowing longer exposure times.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: c j day[SMTP:wa5ekh-at-juno.com] } Sent: Tuesday, May 11, 1999 3:37 PM } To: microscopy-at-sparc5.microscopy.com } Subject: Integrating Digital Cameras? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } I have been looking at some digital cameras for optical } microscopy that "integrate". I would have called it "frame averaging" } or } "pixel averaging", but it appears to be the same image pre-processing } technique. Specifically, I saw an "Optronics" camera and it seemed } to } be able to process low light levels as low as many dark field } situations } without the memory affects seen on low lux type cameras. Has anyone } had } any experience with this particular trade name? I am looking for } service } and dependability technical issues. Also any similar cameras? Please } respond to my email. I don't want to flood the server with this } issue,especially if any comments are slightly critical or } complimentary } and appear commercially biased. I don't mind commercial responses. I } believe they are a real necessary technical component of this and all } issues. Vendors are some of the best and most motivated technical } resources we have available to us.(..but that's another subject...) } } (I have no commercially beneficial interest in any cameras except the } benefits of my laboratory applications, that is I do not sell them. "I } am } not spam."....did I cover this?) } } jeffrey/ 'JD' } } ___________________________________________________________________ } You don't need to buy Internet access to use free Internet e-mail. } Get completely free e-mail from Juno at } http://www.juno.com/getjuno.html } or call Juno at (800) 654-JUNO [654-5866] }
There are a few companies working on making CMOS (rather than = CCD-based) digital cameras which are perfect for the type of integrated solutions = that you describe.=A0 Unfortunately, the technology will only be ready for = the end of this year/beginning of 2000.=A0 If I were you, I would wait before investing, as it is predicted that digital cameras will take a big leap forward.=A0 Our company, Symagery Microsystems, is working on a chip = called the VCA1280.=A0 You can see the specs on our website at = www.symagery.com.=A0 We do not make the cameras themselves, but the following companies are = working on making CMOS cameras using our chip (I hope): Optronics (www.optronics.se), Wintriss, Xillix, SMD, Costar/JAI.
Hope this helps with your research.=A0 If you have any further = questions please do not hesitate to contact me.
-----Original Message----- } From: c j day [ mailto:wa5ekh-at-juno.com {mailto:wa5ekh-at-juno.com} ] Sent: May 11, 1999 5:59 PM To: microscopy-at-Sparc5.Microscopy.Com
Dear DAI Jiyan,
You can measure all three parameters or use accepted values. Cs depends on the polepiece. Nominal values for the CM300 are 0.65mm for the UT lens, 1.2mm for the ST, and 2.0mm for the T lens. Delta depends on Cc, high-voltage ripple and beam energy spread. Nominal values for Cc are 1.5mm UT, 1.5mm ST, and 2.0 for the T lens. You can measure the effective energy spread with a GIF or PEELS. Then a good estimate of delta is given by DELTA (in Angstroms) = 10 x Cc (in mm) x EnergySpread (ppm RMS). The EnergySpread is in parts-per-million (ppm) and is the Root-Mean-Square -- not the Full-Width-at-Half-Maximum value given by the GIF or PEELS measurement. You can use EnergySpread (ppm RMS) = EnergySpread (ppm FWHM)/2.335. For example, if you measure a FWHM energy spread of 1.0eV on a 300keV TEM, it is equivalent to1.0eV/300,000keV = 3.33 ppm FWHM. Then this is equal to 1.43 ppm RMS. Then the delta is 10 x 1.5 x 1.43 = 21.4 Angstrom. Since there is a small contribution from the lens current ripple, the final figure would be about 25 Angstrom. Beam-convergence depends of condenser lens defocus and condenser aperture. Easily measured by viewing the diffraction pattern of a known test structure with the illumination set as for imaging. For our CM300FEG/UT we use Cs = 0.65mm, delta = 25 A, alpha (convergence) = 0.35 milliradian.
A table of Cs and Cc values is given in Ultramicroscopy 47 (1992) 282-297. Measurement of alpha is also explained in Acta Cryst. 31 (1975) 307-310.
A good way to learn how to use image simulation is to attend the NCEM Summer School on Computing for HREM. Go to http://ncem.lbl.gov/ and click on "Summer School".
-Mike
DAI Jiyan wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Dear Microcopists: } I am doing HREM image simulation to compare my HREM image obtained by } Philips EM 300 FEG. The parameters needed for calculation are: } spherical aberration coeffienct (Cs), Gaussin defocus ("DELTA" for the } Defocus spread ), beam convergent semiangle. } I appreciate it if any one can give me the numbers of these three parameters } for EM 300 FEG microscope. Thank you in advance. } } DAI Jiyan } IMRE
Dear colleagues, We want to hire a suitable person for the Central Facility for Electron Microscopy at the Center for Materials Research and Analysis at the University of Nebraska-Lincoln (UNL). The salary will at least likely be in the range of the mid $30k's and is dependent on experience. The medical, dental and retirement benefits package is substantial and includes subsidized tuition for employees to take UNL courses. (BTW, Lincoln is growing, has low unemployment, is great for families, has a good range of live music, dance and theater, and has very good public and other schools.) The Facility provides user-access for ~60 faculty plus their students and other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM and a VG HB501 field-emission STEM, along with a full range of accessories, specimen preparation equipment, computers and darkroom. Some development of new research instrumentation is in progress for mapping of magnetic materials and more development is planned for the Facility. You can find out more about the Facility on our web site (that does need a little work) at URL: http://www.unl.edu/CMRAcfem/ Thanks for passing on the word, Brian Robertson
Analyze/characterize materials using electron microscopy, materials preparation and computer instrumentation. Supervise/train students, faculty and visiting researchers utilizing these methods. Bachelor's with major in physical science, engineering or related field plus three years experience in the operation, repair or design of electron microscopes or other scientific instrumentation. Master's preferred. Must have excellent computer and interpersonal/communication skills. TEM, SEM, x-ray diffraction or materials sample preparation experience preferred. Excellent benefits. Submit cover letter, resume and names, addresses and telephone numbers of three professional references to Professor Brian Robertson, CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position will remain open until a suitable candidate is found. UNL is committed to AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886.
*********************************************************** Assoc. Prof. Brian W. Robertson Department of Mechanical Engineering and Center for Materials Research and Analysis University of Nebraska-Lincoln, N124 WSEC, 17th & Vine Sts., Lincoln, NE 68588-0656, USA ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 **
I don't want to start a thread of "CMOS vs. CCD" here, that probably belongs to a newsgroup or a different listserver, but I want to respond quickly to one posting regarding CMOS cameras. If you want to respond to this posting, please send me email directly and don't respond to the listserver:
CMOS technology is definitely something to watch, it promises higher integration of the sensor and the electronics, which will evetually lead to less expensive cameras. On the other hand, CCD technology has been around for more than 20 years and is a mature technology with proven quality.
CMOS will be targeted first at the consumer market with inexpensive cameras. Whether these cameras and sensors will be sufficient for scientific equipment remains to be seen.
While I think, there is a tremendous potential in CMOS cameras, I personally think that a good CCD camera is the best choice at the moment. If you can afford to wait for a year or two, the situation may change, but for now, I'd go with CCD.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: info-at-soft-imaging.com
} ---------- } From: Marisa Ahmad[SMTP:mahmad-at-semiconductor.com] } Sent: Thursday, May 13, 1999 10:49 AM } To: 'MSA listserver' } Cc: Brigitte Smiley } Subject: FW: Integrating Digital Cameras? } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Jeffrey, } } There are a few companies working on making CMOS (rather than } CCD-based) } digital cameras which are perfect for the type of integrated solutions } that } you describe. Unfortunately, the technology will only be ready for } the end } of this year/beginning of 2000. If I were you, I would wait before } investing, as it is predicted that digital cameras will take a big } leap } forward. Our company, Symagery Microsystems, is working on a chip } called } the VCA1280. You can see the specs on our website at } www.symagery.com. We } do not make the cameras themselves, but the following companies are } working } on making CMOS cameras using our chip (I hope): Optronics } (www.optronics.se), Wintriss, Xillix, SMD, Costar/JAI. } } Hope this helps with your research. If you have any further questions } please do not hesitate to contact me. } } Brigitte Smiley } } brigitte-at-symagery.com {mailto:brigitte-at-symagery.com} } } } } -----Original Message----- } } From: c j day [ mailto:wa5ekh-at-juno.com {mailto:wa5ekh-at-juno.com} ] } Sent: May 11, 1999 5:59 PM } To: microscopy-at-Sparc5.Microscopy.Com } Subject: Integrating Digital Cameras? } } } From: WA5EKH-at-juno.com } } I have been looking at some digital cameras for optical } microscopy that "integrate". I would have called it "frame averaging" } or } "pixel averaging", but it appears to be the same image pre-processing } technique. Specifically, I saw an "Optronics" camera and it seemed } to } be able to process low light levels as low as many dark field } situations } without the memory affects seen on low lux type cameras. Has anyone } had } any experience with this particular trade name? I am looking for } service } and dependability technical issues. Also any similar cameras? Please } respond to my email. I don't want to flood the server with this } issue,especially if any comments are slightly critical or } complimentary } and appear commercially biased. I don't mind commercial responses. I } believe they are a real necessary technical component of this and all } issues. Vendors are some of the best and most motivated technical } resources we have available to us.(..but that's another subject...) } } (I have no commercially beneficial interest in any cameras except the } benefits of my laboratory applications, that is I do not sell them. "I } am } not spam."....did I cover this?) } } jeffrey/ 'JD' } Email: WA5EKH-at-juno.com } } } ___________________________________________________________________ } You don't need to buy Internet access to use free Internet e-mail. } Get completely free e-mail from Juno at } http://www.juno.com/getjuno.html } {http://www.juno.com/getjuno.html} } or call Juno at (800) 654-JUNO [654-5866] } } }
Mid-$30K? Is this for real? Is this the going rate for SEM specialists? That is about 1/4 of what I would consider. There must be some subtle differentiation of what is expected from such positions vs. the qualifications and experience of others in the field. Am I the only one shocked about this?
} JOB ANNOUNCEMENT BELOW -- PLEASE POST } } Dear colleagues, } We want to hire a suitable person for the Central Facility for Electron } Microscopy at the Center for Materials Research and Analysis at the } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } the range of the mid $30k's and is dependent on experience. The medical, } dental and retirement benefits package is substantial and includes } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } growing, has low unemployment, is great for families, has a good range of } live music, dance and theater, and has very good public and other schools.) } The Facility provides user-access for ~60 faculty plus their students and } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } and a VG HB501 field-emission STEM, along with a full range of accessories, } specimen preparation equipment, computers and darkroom. Some development } of new research instrumentation is in progress for mapping of magnetic } materials and more development is planned for the Facility. You can find } out more about the Facility on our web site (that does need a little work) } at URL: http://www.unl.edu/CMRAcfem/ } Thanks for passing on the word, } Brian Robertson } } ********************************************************** } Job Announcement: } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } UNL Center for Materials Research and Analysis } } Analyze/characterize materials using electron microscopy, materials } preparation and computer instrumentation. Supervise/train students, } faculty and visiting researchers utilizing these methods. Bachelor's with } major in physical science, engineering or related field plus three years } experience in the operation, repair or design of electron microscopes or } other scientific instrumentation. Master's preferred. Must have excellent } computer and interpersonal/communication skills. TEM, SEM, x-ray } diffraction or materials sample preparation experience preferred. Excellent } benefits. Submit cover letter, resume and names, addresses and telephone } numbers of three professional references to Professor Brian Robertson, } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } will remain open until a suitable candidate is found. UNL is committed to } AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886. } } } *********************************************************** } Assoc. Prof. Brian W. Robertson } Department of Mechanical Engineering } and Center for Materials Research and Analysis } University of Nebraska-Lincoln, N124 WSEC, } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 **
****************************************************************************** You have been carefully selected to receive the following as a person interested in opportunities based upon your previous Internet postings or visits to one of our affiliate web sites. If you have received this message in error, please accept our apology as a responsible e-mailer. However, this is a ONE-TIME only announcement and is not intended as a SPAM letter, therefore you need not reply. You are automatically moved to our remove list. Again, if you are not interested, we sincerely apologize for the inconvenience. Thank you. ******************************************************************************
Dear Friend,
You can earn $50,000 or more in next the 90 days sending e-mail. Seem impossible? Read on for details.
"AS SEEN ON NATIONAL TV"
Thank you for your time and interest. This is the letter you've been reading about in the news lately. Due to the popularity of this letter on the Internet, a major nightly news program recently devoted an entire show to the investigation of the program described below to see if it really can make people money.
The show also investigated whether or not the program was legal. Their findings proved once and for all that there are absolutely no laws prohibiting the participation in the program. This has helped to show people that this is a simple, harmless and fun way to make some extra money at home.
The results of this show have been truly remarkable. So many people are participating that those involved are doing much better than ever before. Since everyone makes more as more people try it out, its been very exciting to be a part of lately. You will understand once you experience it.
HERE IT IS BELOW:
*** Print This Now For Future Reference ***
The following income opportunity is one you may be interested in taking a look at. It can be started with VERY LITTLE investment and the income return is TREMENDOUS!!!
$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ If you would like to make at least $50,000 in less than 90 days ! Please read the enclosed program...THEN READ IT AGAIN!!! $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$
THIS IS A LEGITIMATE, LEGAL, MONEY MAKING OPPORTUNITY.It does not require you to come into contact with people, do any hard work, and best of all, you never have to leave the house except to get the mail. If you believe that someday you'll get that big break that you 'vebeen waiting for, THIS IS IT! Simply follow the instructions, andyour dreams will come true. This multi-level e-mail order marketingprogram works perfectly...100% EVERY TIME.
E-mail is the sales tool of the future. Take advantage of this non-commercialized method of advertising NOW!!! The longer you wait, the more people will be doing business using e-mail. Get your piece of this action!!!
MULTI-LEVEL MARKETING (MLM) has finally gained respectability. It is being taught in the Harvard Business School, and both Stanford Research and the Wall Street Journal have stated that between 50% and 65% of all goods and services will be sold through multi-level methods by the mid to late 1990's. This is a Multi-Billion Dollar industry and of the 500,000 millionaires in the U.S., 20% (100,000) made their fortune in the last several years in MLM. Moreover, statistics show 45 people become millionaires everyday through Multi-Level Marketing.
You may have heard this story before, but over the summer Donald Trump made an appearance on the David Letterman show. Dave asked him what he would do if he lost everything and had to start over from scratch. Without hesitating, Trump said he would find a good network marketing company and get to work. The audience started to hoot and boo him. He looked out at the audience and dead-panned his response: "That's why I'm sitting up here and you are all sitting out there!"
The enclosed information is something I almost let slip through my fingers. Fortunately, sometime later I re-read everything and gave somethought and study to it. My name is Johnathon Rourke. Two years ago, the corporation I worked at for the past twelve years down-sized and my position was eliminated. After unproductive job interviews, I decided to open my own business. Over the past year, I incurred many unforeseen financial problems. I owed my family, friends and creditors over $35,000. The economy was taking a toll on my business and I just couldn't seem to make ends meet. I had to refinance and borrow against my home to support my family and struggling business. AT THAT MOMENT something significant happened in my life and I am writing to share the experience in hopes that this will change your life FOREVER FINANCIALLY!!!
In mid December, I received this program via e-mail. Six month's prior to receiving this program I had been sending away for information on various business opportunities. All of the programs I received, in my opinion, were not cost effective. They were either too difficult for me to comprehend or the initial investment was too much for me to risk to see if they would work or not. One claimed that I would make a million dollars in one year...it didn't tell me I'd have to write a book to make it!
But like I was saying, in December of 1997 I received this program. I didn't send for it, or ask for it, they just got my name off a mailing list.THANK GOODNESS FOR THAT!!! After reading it several times, to make sure I was reading it correctly, I couldn't believe my eyes. Here was a MONEY MAKING PHENOMENON. I could invest as much as I wanted to start, without putting me further into debt. After I got a pencil and paper and figured it out, I would at least get my money back. But like most of you I was still a little skeptical and a little worried about the legal aspects of it all. So I checked it out with the U.S. Post Office (1-800-725-2161 24-hrs) and they confirmed that it is indeed legal! After determining the program was LEGAL and NOT A CHAIN LETTER, I decided "WHY NOT."
Initially I sent out 10,000 e-mails. It cost me about $15 for my time on-line. The great thing about e-mail is that I don't need any money for printing to send out the program, and because all of my orders are fulfilled via e-mail, my only expense is my time. I am telling you like it is I hope it doesn't turn you off, but I promised myself that I would not "rip-off" anyone, no matter how much money it made me.
In less than one week, I was starting to receive orders for REPORT #1 By January 13, I had received 26 orders for REPORT #1. Your goal is to "RECEIVE at least 20 ORDERS FOR REPORT #1 WITHIN 2 WEEKS. IF YOU DON'T, SEND OUT MORE PROGRAMS UNTIL YOU DO!" My first step in making $50,000 in 90 days was done. By January 30, I had received 196 orders for REPORT #2. Your goal is to "RECEIVE AT LEAST 100+ ORDERS FOR REPORT #2 WITHIN 2 WEEKS. IF NOT, SEND OUT MORE PROGRAMS UNTIL YOU DO. ONCE YOU HAVE 100 ORDERS, THE REST IS EASY, RELAX, YOU WILL MAKE YOUR $50,000 GOAL." Well, I had 196 orders for REPORT #2, 96 more than I needed. So I sat back and relaxed. By March 1, of my e-mailing of 10,000, I received $58,000 with more coming in every day.
I paid off ALL my debts and bought a much needed new car. Please take time to read the attached program, IT WILL CHANGE YOUR LIFE FOREVER!! ! Remember, it won't work if you don't try it. This program does work , but you must follow it EXACTLY! Especially the rules of not trying to place your name in a different place. It won't work and you'll lose out on a lot of money! In order for this program to work, you must meet your goal of 20+ orders for REPORT #1, and 100+ orders for REPORT #2 and you will make $50,000 or more in 90 days. I AM LIVING PROOF THAT IT WORKS!!!
If you choose not to participate in this program, I am sorry. It really is a great opportunity with little cost or risk to you. If you choose to participate, follow the program and you will be on your way to financial security. If you are a fellow business owner and are in financial trouble like I was, or you want to start your own business, consider this a sign. I DID!
Sincerely, Johnathon Rourke
A PERSONAL NOTE FROM THE ORIGINATOR OF THIS PROGRAM:
By the time you have read the enclosed program and reports, you should have concluded that such a program, and one that is legal, could not have been created by an amateur.
Let me tell you a little about myself. I had a profitable business for 10 years. Then in 1979 my business began falling off. I was doing the same things that were previously successful for me, but it wasn't working. Finally, I figured it out. It wasn't me, it was the economy. Inflation and recession had replaced the stable economy that had been with us since 1945.I don't have to tell you what happened to the unemployment rate... because many of you know from first hand experience. There were more failures and bankruptcies than ever before.
The middle class was vanishing. Those who knew what they were doing invested wisely and moved up. Those who did not, including those who never had anything to save or invest, were moving down into the ranks of the poor. As the saying goes, "THE RICH GET RICHER AND THE POOR GET POORER." The traditional methods of making money will never allow you to "move up" or "get rich", inflation will see to that.
You have just received information that can give you financial freedom for the rest of your life, with "NO RISK" and "JUST A LITTLE BIT OF EFFORT." You can make more money in the next few months than you have ever imagined. I should also point out that I will not see a penny of this money, nor anyone else who has provided a testimonial for this program. I have already made over 4 MILLION DOLLARS!I have retired from the program after sending thousands and thousands of programs.
Follow the program EXACTLY AS INSTRUCTED. Do not change it in any way . It works exceedingly well as it is now. Remember to e-mail a copy of this exciting report to everyone you can think of. One of the people you send this to may send out 50,000...and your name will be on everyone of them!
Remember though, the more you send out the more potential customers you will reach.
So my friend, I have given you the ideas, information, materials and opportunity to become financially independent. IT IS UP TO YOU NOW!
"THINK ABOUT IT"
Before you delete this program from your mailbox, as I almost did, take a little time to read it and REALLY THINK ABOUT IT. Get a pencil and figure out what could happen when YOU participate. Figure out the worst possible response and no matter how you calculate it, you will still make a lot of money! You will definitely get back what you invested. Any doubts you have will vanish when your first orders come in. IT WORKS!
Jody Jacobs, Richmond, VA
HERE'S HOW THIS AMAZING PROGRAM WILL MAKE YOU THOUSANDS OF DOLLAR$
INSTRUCTIONS:
This method of raising capital REALLY WORKS 100% EVERY TIME. I am sure that you could use up to $50,000 or more in the next 90 days. Before you say "BULL... ", please read this program carefully.
This is not a chain letter, but a perfectly legal money making opportunity. Basically, this is what you do: As with all multi-level businesses, we build our business by recruiting new partners and selling our products. Every state in the USA allows you to recruit new multi-level business partners, and we offer a product for EVERY dollar sent. YOUR ORDERS COME BY MAIL AND ARE FILLED BY E-MAIL, so you are not involved in personal selling. You do it privately in your own home, store or office. This is the GREATEST Multi-Level Mail Order Marketing anywhere.
This is what you MUST do:
1. Order all 4 reports shown on the list below (you can't sell them if youdon't order them). -- For each report, send $5.00 CASH, the NAME & NUMBER OF THE REPORT YOU ARE ORDERING, YOUR E-MAIL ADDRESS, and YOUR NAME & RETURN ADDRESS (in case of a problem) to the person whose name appears on the list next to the report. MAKE SURE YOUR RETURN ADDRESS IS ON YOUR ENVELOPE IN CASE OF ANY MAIL PROBLEMS! -- When you place your order, make sure you order each of the four reports. You will need all four reports so that you can save them on your computer and resell them. -- Within a few days you will receive, via e-mail, each of the four reports. Save them on your computer so they will be accessible for you to send to the 1,000's of people who will order them from you.
2. IMPORTANT DO NOT alter the names of the people who are listed next to each report, or their sequence on the list, in any way other than is instructed below in steps "a" through "f" or you will lose out on the majority of your profits. Once you understand the way this works, you'll also see how it doesn't work if you change it. Remember, this method has been tested,and if you alter it, it will not work. a. Look below for the listing of available reports. b. After you've ordered the four reports, take this advertisement and remove the name and address under REPORT #4. This person has made it through the cycle and is no doubt counting their $50,000! c. Move the name and address under REPORT #3 down to REPORT #4. d. Move the name and address under REPORT #2 down to REPORT #3. e. Move the name and address under REPORT #1 down to REPORT #2. f. Insert your name/address in the REPORT #1 position.
Please make sure you COPY ALL INFORMATION, every name and address, ACCURATELY!
3. Take this entire letter, including the modified list of names, and save it to your computer. Make NO changes to the instruction portion of this letter.
Your cost to participate in this is practically nothing (surely you can afford $20). You obviously already have an Internet connection and e-mail is FREE!
There are two primary methods of building your downline:
METHOD #1: SENDING BULK E-MAIL
Let's say that you decide to start small, just to see how it goes, and we'll assume you and all those involved send out only 2,000 programs each. Let's also assume that the mailing receives a 0.5% response. Using a good list the response could be much better. Also, many people will send out hundreds of thousands of programs instead of 2,000. But continuing with this example, you send out only 2,000 programs. With a 0.5% response, that is only 10 orders for REPORT #1. Those 10 people respond by sending out 2,000 programs each for a total of 20,000. Out of those 0.5%, 100 people respond and order REPORT #2. Those 100 mail out 2,000 programs each for a total of 200,000. The 0.5% response to that is 1,000 orders for REPORT #3. Those 1,000 send out 2,000 programs each for a 2,000,000 total. The 0.5% response to that is 10,000 orders for REPORT #4. That's 10,000 $5 bills for you. CASH!!! Your total income in this example is $50 + $500 + $5,000 + $50,000 for a total of $55,550!!! REMEMBER FRIEND, THIS IS ASSUMING 1,990 OUT OF THE 2,000 PEOPLE YOU MAIL TO WILL DO ABSOLUTELY NOTHING AND TRASH THIS PROGRAM! DARE TO THINK FOR A MOMENT WHAT WOULD HAPPEN IF EVERYONE, OR HALF SENT OUT 100,000 PROGRAMS INSTEAD OF 2,000. Believe me, many people will do justthat, and more! By the way, your cost to participate in this is practically nothing. You obviously already have an Internet connection and e-mail is FREE!!! REPORT #2 will show you the best methods for bulk e-mailing, tell you where to obtain free bulk e-mail software and where to obtain e-mail lists.
METHOD #2 - PLACING FREE ADS ON THE INTERNET
Advertising on the internet is very, very inexpensive, and there are HUNDREDS of FREE places to advertise. Let's say you decide to start small just to see how well it works. Assume your goal is to get ONLY 10 people to participate on your first level. (Placing a lot of FREE ads on the Internet will EASILY get a larger response.) Also assume that everyone else in YOUR ORGANIZATION gets ONLY 10 downline members. Follow this example to achieve the STAGGERING results below:
1st level--your 10 members with $5.......................................$50 2nd level--10 members from those 10 ($5 x 100)..................$500 3rd level--10 members from those 100 ($5 x 1,000)...........$5,000 4th level--10 members from those 1,000 ($5 x 10,000).....$50,000 THIS TOTALS ----------} $55,550
Remember friends, this assumes that the people who participate only recruit 10 people each. Think for a moment what would happen if they got 20 people to participate! Most people get 100's of participants! THINK ABOUT IT! For every $5.00 you receive, all you must do is e-mail them the report they ordered. THAT'S IT! ALWAYS PROVIDE SAME-DAY SERVICE ON ALL ORDERS! This will guarantee that the e-mail THEY send out with YOUR name and address on it will be prompt because they can't advertise until they receive the report!
AVAILABLE REPORTS
*** Order Each REPORT by NUMBER and NAME *** Notes: -- ALWAYS SEND $5 CASH (U.S. CURRENCY) FOR EACH REPORT. CHECKS NOT ACCEPTED. -- ALWAYS SEND YOUR ORDER VIA FIRST CLASS MAIL. -- Make sure the cash is concealed by wrapping it in at least two sheets of paper. On one of those sheets of paper, include: (a) the number & name of the report you are ordering, (b) your e-mail address, and (c) your name & postal address.
PLACE YOUR ORDER FOR THESE REPORTS NOW:
REPORT #1
----------------------------------------------------------------- "The Insider's Guide to Advertising for Free on the Internet" ORDER REPORT #1 FROM: J. Grainger Via del Perugino, 5 40139 Bologna, Italy ----------------------------------------------------------------- REPORT #2 "The Insider's Guide to Sending Bulk E-mail on the Internet" ORDER REPORT #2 FROM: D. Clark P.O. Box 685 Grangeville, Idaho 83530 ----------------------------------------------------------------- REPORT #3 "The Secrets to Multilevel Marketing on the Internet" ORDER REPORT #3 FROM: Peter Gooijer Galileistraat 33 5621 AE Eindhoven The Netherlands ----------------------------------------------------------------- REPORT #4 "How to become a Millionaire utilizing the Power of Multilevel Marketing and the Internet" ORDER REPORT #4 FROM: J. Russ 8034 Linda Vista Rd. # 1M San Diego, Ca 92111 ----------------------------------------------------------------
About 50,000 new people get online every month!
******* TIPS FOR SUCCESS ******* -- TREAT THIS AS YOUR BUSINESS! Be prompt, professional, and follow the directions accurately. -- Send for the four reports IMMEDIATELY so you will have them when the orders start coming in because: When you receive a $5 order, you MUST send out the requested product/report. -- ALWAYS PROVIDE SAME-DAY SERVICE ON THE ORDERS YOU RECEIVE. -- Be patient and persistent with this program. If you follow the instructions exactly, your results WILL BE SUCCESSFUL! -- ABOVE ALL, HAVE FAITH IN YOURSELF AND KNOW YOU WILL SUCCEED!
******* YOUR SUCCESS GUIDELINES ******* Follow these guidelines to guarantee your success:
If you don't receive 20 orders for REPORT #1 within two weeks, continue
advertising or sending e-mails until you do. Then, a couple of weeks later you should receive at least 100 orders for REPORT#2. If you don 't, continue advertising or sending e-mails until you do. Once you have received 100 or more orders for REPORT #2, YOU CAN RELAX, because the system is already working for you, and the cash will continue to roll in!
THIS IS IMPORTANT TO REMEMBER: Every time your name is moved down on the list, you are placed in front of a DIFFERENT report. You can KEEP TRACK of your PROGRESS by watching which report people are ordering from you. If you want to generate more income, send another batch of e-mails or continue placing ads and start the whole process again! There is no limit to the income you will generate from this business!
Before you make your decision as to whether or not you participate in this program. Please answer one question. DO YOU WANT TO CHANGE YOUR LIFE? If the answer is yes, please look at the following facts about this program:
1. You are selling a product which does not Cost anything to PRODUCE, SHIP OR ADVERTISE. 2. All of your customers pay you in CASH! 3. E-mail is without question the most powerful method of distributing information on earth. This program combines the distribution power of e-mail together with the revenue generating power of multi-level marketing. 4. Your only expense--other than your initial $20 investment--is your time! 5. Virtually all of the income you generate from this program is PURE PROFIT! 6. This program will change your LIFE FOREVER.
ACT NOW!Take your first step toward achieving financial independence. Orderthe reports and follow the program outlined above--SUCCESSwill be yourreward.
Thank you for your time and consideration.
PLEASE NOTE: If you need help with starting a business, registering a business name, learning how income tax is handled, etc., contact your localoffice of the Small Business Administration (a Federal Agency) 1-800-827-5722 for free help and answers to questions. Also, the InternalRevenue Service offers free help via telephone and free seminars aboutbusiness tax requirements. Your earnings are highly dependant on youractivities and advertising. The information contained on this site and in the report constitutes no guarantees stated nor implied. In the event that it is determined that this site or report constitutes a guarantee of any kind, that guarantee is now void. The earnings amounts listed on this site and in the report are estimates only. If you have any questions of the legality of this program, contact the Office of Associate Director for Marketing Practices, Federal Trade Commission, Bureau of Consumer Protection in Washington, DC.
///////////////////////////////////////////////////////////////// Remove at rhobbs12-at-yahoo.com /////////////////////////////////////////////////////////////////
****************************************************************************** You have been carefully selected to receive the following as a person interested in opportunities based upon your previous Internet postings or visits to one of our affiliate web sites. If you have received this message in error, please accept our apology as a responsible e-mailer. However, this is a ONE-TIME only announcement and is not intended as a SPAM letter, therefore you need not reply. You are automatically moved to our remove list. Again, if you are not interested, we sincerely apologize for the inconvenience. Thank you. ******************************************************************************
Dear Friend,
You can earn $50,000 or more in next the 90 days sending e-mail. Seem impossible? Read on for details.
"AS SEEN ON NATIONAL TV"
Thank you for your time and interest. This is the letter you've been reading about in the news lately. Due to the popularity of this letter on the Internet, a major nightly news program recently devoted an entire show to the investigation of the program described below to see if it really can make people money.
The show also investigated whether or not the program was legal. Their findings proved once and for all that there are absolutely no laws prohibiting the participation in the program. This has helped to show people that this is a simple, harmless and fun way to make some extra money at home.
The results of this show have been truly remarkable. So many people are participating that those involved are doing much better than ever before. Since everyone makes more as more people try it out, its been very exciting to be a part of lately. You will understand once you experience it.
HERE IT IS BELOW:
*** Print This Now For Future Reference ***
The following income opportunity is one you may be interested in taking a look at. It can be started with VERY LITTLE investment and the income return is TREMENDOUS!!!
$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ If you would like to make at least $50,000 in less than 90 days ! Please read the enclosed program...THEN READ IT AGAIN!!! $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$
THIS IS A LEGITIMATE, LEGAL, MONEY MAKING OPPORTUNITY.It does not require you to come into contact with people, do any hard work, and best of all, you never have to leave the house except to get the mail. If you believe that someday you'll get that big break that you 'vebeen waiting for, THIS IS IT! Simply follow the instructions, andyour dreams will come true. This multi-level e-mail order marketingprogram works perfectly...100% EVERY TIME.
E-mail is the sales tool of the future. Take advantage of this non-commercialized method of advertising NOW!!! The longer you wait, the more people will be doing business using e-mail. Get your piece of this action!!!
MULTI-LEVEL MARKETING (MLM) has finally gained respectability. It is being taught in the Harvard Business School, and both Stanford Research and the Wall Street Journal have stated that between 50% and 65% of all goods and services will be sold through multi-level methods by the mid to late 1990's. This is a Multi-Billion Dollar industry and of the 500,000 millionaires in the U.S., 20% (100,000) made their fortune in the last several years in MLM. Moreover, statistics show 45 people become millionaires everyday through Multi-Level Marketing.
You may have heard this story before, but over the summer Donald Trump made an appearance on the David Letterman show. Dave asked him what he would do if he lost everything and had to start over from scratch. Without hesitating, Trump said he would find a good network marketing company and get to work. The audience started to hoot and boo him. He looked out at the audience and dead-panned his response: "That's why I'm sitting up here and you are all sitting out there!"
The enclosed information is something I almost let slip through my fingers. Fortunately, sometime later I re-read everything and gave somethought and study to it. My name is Johnathon Rourke. Two years ago, the corporation I worked at for the past twelve years down-sized and my position was eliminated. After unproductive job interviews, I decided to open my own business. Over the past year, I incurred many unforeseen financial problems. I owed my family, friends and creditors over $35,000. The economy was taking a toll on my business and I just couldn't seem to make ends meet. I had to refinance and borrow against my home to support my family and struggling business. AT THAT MOMENT something significant happened in my life and I am writing to share the experience in hopes that this will change your life FOREVER FINANCIALLY!!!
In mid December, I received this program via e-mail. Six month's prior to receiving this program I had been sending away for information on various business opportunities. All of the programs I received, in my opinion, were not cost effective. They were either too difficult for me to comprehend or the initial investment was too much for me to risk to see if they would work or not. One claimed that I would make a million dollars in one year...it didn't tell me I'd have to write a book to make it!
But like I was saying, in December of 1997 I received this program. I didn't send for it, or ask for it, they just got my name off a mailing list.THANK GOODNESS FOR THAT!!! After reading it several times, to make sure I was reading it correctly, I couldn't believe my eyes. Here was a MONEY MAKING PHENOMENON. I could invest as much as I wanted to start, without putting me further into debt. After I got a pencil and paper and figured it out, I would at least get my money back. But like most of you I was still a little skeptical and a little worried about the legal aspects of it all. So I checked it out with the U.S. Post Office (1-800-725-2161 24-hrs) and they confirmed that it is indeed legal! After determining the program was LEGAL and NOT A CHAIN LETTER, I decided "WHY NOT."
Initially I sent out 10,000 e-mails. It cost me about $15 for my time on-line. The great thing about e-mail is that I don't need any money for printing to send out the program, and because all of my orders are fulfilled via e-mail, my only expense is my time. I am telling you like it is I hope it doesn't turn you off, but I promised myself that I would not "rip-off" anyone, no matter how much money it made me.
In less than one week, I was starting to receive orders for REPORT #1 By January 13, I had received 26 orders for REPORT #1. Your goal is to "RECEIVE at least 20 ORDERS FOR REPORT #1 WITHIN 2 WEEKS. IF YOU DON'T, SEND OUT MORE PROGRAMS UNTIL YOU DO!" My first step in making $50,000 in 90 days was done. By January 30, I had received 196 orders for REPORT #2. Your goal is to "RECEIVE AT LEAST 100+ ORDERS FOR REPORT #2 WITHIN 2 WEEKS. IF NOT, SEND OUT MORE PROGRAMS UNTIL YOU DO. ONCE YOU HAVE 100 ORDERS, THE REST IS EASY, RELAX, YOU WILL MAKE YOUR $50,000 GOAL." Well, I had 196 orders for REPORT #2, 96 more than I needed. So I sat back and relaxed. By March 1, of my e-mailing of 10,000, I received $58,000 with more coming in every day.
I paid off ALL my debts and bought a much needed new car. Please take time to read the attached program, IT WILL CHANGE YOUR LIFE FOREVER!! ! Remember, it won't work if you don't try it. This program does work , but you must follow it EXACTLY! Especially the rules of not trying to place your name in a different place. It won't work and you'll lose out on a lot of money! In order for this program to work, you must meet your goal of 20+ orders for REPORT #1, and 100+ orders for REPORT #2 and you will make $50,000 or more in 90 days. I AM LIVING PROOF THAT IT WORKS!!!
If you choose not to participate in this program, I am sorry. It really is a great opportunity with little cost or risk to you. If you choose to participate, follow the program and you will be on your way to financial security. If you are a fellow business owner and are in financial trouble like I was, or you want to start your own business, consider this a sign. I DID!
Sincerely, Johnathon Rourke
A PERSONAL NOTE FROM THE ORIGINATOR OF THIS PROGRAM:
By the time you have read the enclosed program and reports, you should have concluded that such a program, and one that is legal, could not have been created by an amateur.
Let me tell you a little about myself. I had a profitable business for 10 years. Then in 1979 my business began falling off. I was doing the same things that were previously successful for me, but it wasn't working. Finally, I figured it out. It wasn't me, it was the economy. Inflation and recession had replaced the stable economy that had been with us since 1945.I don't have to tell you what happened to the unemployment rate... because many of you know from first hand experience. There were more failures and bankruptcies than ever before.
The middle class was vanishing. Those who knew what they were doing invested wisely and moved up. Those who did not, including those who never had anything to save or invest, were moving down into the ranks of the poor. As the saying goes, "THE RICH GET RICHER AND THE POOR GET POORER." The traditional methods of making money will never allow you to "move up" or "get rich", inflation will see to that.
You have just received information that can give you financial freedom for the rest of your life, with "NO RISK" and "JUST A LITTLE BIT OF EFFORT." You can make more money in the next few months than you have ever imagined. I should also point out that I will not see a penny of this money, nor anyone else who has provided a testimonial for this program. I have already made over 4 MILLION DOLLARS!I have retired from the program after sending thousands and thousands of programs.
Follow the program EXACTLY AS INSTRUCTED. Do not change it in any way . It works exceedingly well as it is now. Remember to e-mail a copy of this exciting report to everyone you can think of. One of the people you send this to may send out 50,000...and your name will be on everyone of them!
Remember though, the more you send out the more potential customers you will reach.
So my friend, I have given you the ideas, information, materials and opportunity to become financially independent. IT IS UP TO YOU NOW!
"THINK ABOUT IT"
Before you delete this program from your mailbox, as I almost did, take a little time to read it and REALLY THINK ABOUT IT. Get a pencil and figure out what could happen when YOU participate. Figure out the worst possible response and no matter how you calculate it, you will still make a lot of money! You will definitely get back what you invested. Any doubts you have will vanish when your first orders come in. IT WORKS!
Jody Jacobs, Richmond, VA
HERE'S HOW THIS AMAZING PROGRAM WILL MAKE YOU THOUSANDS OF DOLLAR$
INSTRUCTIONS:
This method of raising capital REALLY WORKS 100% EVERY TIME. I am sure that you could use up to $50,000 or more in the next 90 days. Before you say "BULL... ", please read this program carefully.
This is not a chain letter, but a perfectly legal money making opportunity. Basically, this is what you do: As with all multi-level businesses, we build our business by recruiting new partners and selling our products. Every state in the USA allows you to recruit new multi-level business partners, and we offer a product for EVERY dollar sent. YOUR ORDERS COME BY MAIL AND ARE FILLED BY E-MAIL, so you are not involved in personal selling. You do it privately in your own home, store or office. This is the GREATEST Multi-Level Mail Order Marketing anywhere.
This is what you MUST do:
1. Order all 4 reports shown on the list below (you can't sell them if youdon't order them). -- For each report, send $5.00 CASH, the NAME & NUMBER OF THE REPORT YOU ARE ORDERING, YOUR E-MAIL ADDRESS, and YOUR NAME & RETURN ADDRESS (in case of a problem) to the person whose name appears on the list next to the report. MAKE SURE YOUR RETURN ADDRESS IS ON YOUR ENVELOPE IN CASE OF ANY MAIL PROBLEMS! -- When you place your order, make sure you order each of the four reports. You will need all four reports so that you can save them on your computer and resell them. -- Within a few days you will receive, via e-mail, each of the four reports. Save them on your computer so they will be accessible for you to send to the 1,000's of people who will order them from you.
2. IMPORTANT DO NOT alter the names of the people who are listed next to each report, or their sequence on the list, in any way other than is instructed below in steps "a" through "f" or you will lose out on the majority of your profits. Once you understand the way this works, you'll also see how it doesn't work if you change it. Remember, this method has been tested,and if you alter it, it will not work. a. Look below for the listing of available reports. b. After you've ordered the four reports, take this advertisement and remove the name and address under REPORT #4. This person has made it through the cycle and is no doubt counting their $50,000! c. Move the name and address under REPORT #3 down to REPORT #4. d. Move the name and address under REPORT #2 down to REPORT #3. e. Move the name and address under REPORT #1 down to REPORT #2. f. Insert your name/address in the REPORT #1 position.
Please make sure you COPY ALL INFORMATION, every name and address, ACCURATELY!
3. Take this entire letter, including the modified list of names, and save it to your computer. Make NO changes to the instruction portion of this letter.
Your cost to participate in this is practically nothing (surely you can afford $20). You obviously already have an Internet connection and e-mail is FREE!
There are two primary methods of building your downline:
METHOD #1: SENDING BULK E-MAIL
Let's say that you decide to start small, just to see how it goes, and we'll assume you and all those involved send out only 2,000 programs each. Let's also assume that the mailing receives a 0.5% response. Using a good list the response could be much better. Also, many people will send out hundreds of thousands of programs instead of 2,000. But continuing with this example, you send out only 2,000 programs. With a 0.5% response, that is only 10 orders for REPORT #1. Those 10 people respond by sending out 2,000 programs each for a total of 20,000. Out of those 0.5%, 100 people respond and order REPORT #2. Those 100 mail out 2,000 programs each for a total of 200,000. The 0.5% response to that is 1,000 orders for REPORT #3. Those 1,000 send out 2,000 programs each for a 2,000,000 total. The 0.5% response to that is 10,000 orders for REPORT #4. That's 10,000 $5 bills for you. CASH!!! Your total income in this example is $50 + $500 + $5,000 + $50,000 for a total of $55,550!!! REMEMBER FRIEND, THIS IS ASSUMING 1,990 OUT OF THE 2,000 PEOPLE YOU MAIL TO WILL DO ABSOLUTELY NOTHING AND TRASH THIS PROGRAM! DARE TO THINK FOR A MOMENT WHAT WOULD HAPPEN IF EVERYONE, OR HALF SENT OUT 100,000 PROGRAMS INSTEAD OF 2,000. Believe me, many people will do justthat, and more! By the way, your cost to participate in this is practically nothing. You obviously already have an Internet connection and e-mail is FREE!!! REPORT #2 will show you the best methods for bulk e-mailing, tell you where to obtain free bulk e-mail software and where to obtain e-mail lists.
METHOD #2 - PLACING FREE ADS ON THE INTERNET
Advertising on the internet is very, very inexpensive, and there are HUNDREDS of FREE places to advertise. Let's say you decide to start small just to see how well it works. Assume your goal is to get ONLY 10 people to participate on your first level. (Placing a lot of FREE ads on the Internet will EASILY get a larger response.) Also assume that everyone else in YOUR ORGANIZATION gets ONLY 10 downline members. Follow this example to achieve the STAGGERING results below:
1st level--your 10 members with $5.......................................$50 2nd level--10 members from those 10 ($5 x 100)..................$500 3rd level--10 members from those 100 ($5 x 1,000)...........$5,000 4th level--10 members from those 1,000 ($5 x 10,000).....$50,000 THIS TOTALS ----------} $55,550
Remember friends, this assumes that the people who participate only recruit 10 people each. Think for a moment what would happen if they got 20 people to participate! Most people get 100's of participants! THINK ABOUT IT! For every $5.00 you receive, all you must do is e-mail them the report they ordered. THAT'S IT! ALWAYS PROVIDE SAME-DAY SERVICE ON ALL ORDERS! This will guarantee that the e-mail THEY send out with YOUR name and address on it will be prompt because they can't advertise until they receive the report!
AVAILABLE REPORTS
*** Order Each REPORT by NUMBER and NAME *** Notes: -- ALWAYS SEND $5 CASH (U.S. CURRENCY) FOR EACH REPORT. CHECKS NOT ACCEPTED. -- ALWAYS SEND YOUR ORDER VIA FIRST CLASS MAIL. -- Make sure the cash is concealed by wrapping it in at least two sheets of paper. On one of those sheets of paper, include: (a) the number & name of the report you are ordering, (b) your e-mail address, and (c) your name & postal address.
PLACE YOUR ORDER FOR THESE REPORTS NOW:
REPORT #1
----------------------------------------------------------------- "The Insider's Guide to Advertising for Free on the Internet" ORDER REPORT #1 FROM: J. Grainger Via del Perugino, 5 40139 Bologna, Italy ----------------------------------------------------------------- REPORT #2 "The Insider's Guide to Sending Bulk E-mail on the Internet" ORDER REPORT #2 FROM: D. Clark P.O. Box 685 Grangeville, Idaho 83530 ----------------------------------------------------------------- REPORT #3 "The Secrets to Multilevel Marketing on the Internet" ORDER REPORT #3 FROM: Peter Gooijer Galileistraat 33 5621 AE Eindhoven The Netherlands ----------------------------------------------------------------- REPORT #4 "How to become a Millionaire utilizing the Power of Multilevel Marketing and the Internet" ORDER REPORT #4 FROM: J. Russ 8034 Linda Vista Rd. # 1M San Diego, Ca 92111 ----------------------------------------------------------------
About 50,000 new people get online every month!
******* TIPS FOR SUCCESS ******* -- TREAT THIS AS YOUR BUSINESS! Be prompt, professional, and follow the directions accurately. -- Send for the four reports IMMEDIATELY so you will have them when the orders start coming in because: When you receive a $5 order, you MUST send out the requested product/report. -- ALWAYS PROVIDE SAME-DAY SERVICE ON THE ORDERS YOU RECEIVE. -- Be patient and persistent with this program. If you follow the instructions exactly, your results WILL BE SUCCESSFUL! -- ABOVE ALL, HAVE FAITH IN YOURSELF AND KNOW YOU WILL SUCCEED!
******* YOUR SUCCESS GUIDELINES ******* Follow these guidelines to guarantee your success:
If you don't receive 20 orders for REPORT #1 within two weeks, continue
advertising or sending e-mails until you do. Then, a couple of weeks later you should receive at least 100 orders for REPORT#2. If you don 't, continue advertising or sending e-mails until you do. Once you have received 100 or more orders for REPORT #2, YOU CAN RELAX, because the system is already working for you, and the cash will continue to roll in!
THIS IS IMPORTANT TO REMEMBER: Every time your name is moved down on the list, you are placed in front of a DIFFERENT report. You can KEEP TRACK of your PROGRESS by watching which report people are ordering from you. If you want to generate more income, send another batch of e-mails or continue placing ads and start the whole process again! There is no limit to the income you will generate from this business!
Before you make your decision as to whether or not you participate in this program. Please answer one question. DO YOU WANT TO CHANGE YOUR LIFE? If the answer is yes, please look at the following facts about this program:
1. You are selling a product which does not Cost anything to PRODUCE, SHIP OR ADVERTISE. 2. All of your customers pay you in CASH! 3. E-mail is without question the most powerful method of distributing information on earth. This program combines the distribution power of e-mail together with the revenue generating power of multi-level marketing. 4. Your only expense--other than your initial $20 investment--is your time! 5. Virtually all of the income you generate from this program is PURE PROFIT! 6. This program will change your LIFE FOREVER.
ACT NOW!Take your first step toward achieving financial independence. Orderthe reports and follow the program outlined above--SUCCESSwill be yourreward.
Thank you for your time and consideration.
PLEASE NOTE: If you need help with starting a business, registering a business name, learning how income tax is handled, etc., contact your localoffice of the Small Business Administration (a Federal Agency) 1-800-827-5722 for free help and answers to questions. Also, the InternalRevenue Service offers free help via telephone and free seminars aboutbusiness tax requirements. Your earnings are highly dependant on youractivities and advertising. The information contained on this site and in the report constitutes no guarantees stated nor implied. In the event that it is determined that this site or report constitutes a guarantee of any kind, that guarantee is now void. The earnings amounts listed on this site and in the report are estimates only. If you have any questions of the legality of this program, contact the Office of Associate Director for Marketing Practices, Federal Trade Commission, Bureau of Consumer Protection in Washington, DC.
///////////////////////////////////////////////////////////////// Remove at rhobbs12-at-yahoo.com /////////////////////////////////////////////////////////////////
****************************************************************************** You have been carefully selected to receive the following as a person interested in opportunities based upon your previous Internet postings or visits to one of our affiliate web sites. If you have received this message in error, please accept our apology as a responsible e-mailer. However, this is a ONE-TIME only announcement and is not intended as a SPAM letter, therefore you need not reply. You are automatically moved to our remove list. Again, if you are not interested, we sincerely apologize for the inconvenience. Thank you. ******************************************************************************
Dear Friend,
You can earn $50,000 or more in next the 90 days sending e-mail. Seem impossible? Read on for details.
"AS SEEN ON NATIONAL TV"
Thank you for your time and interest. This is the letter you've been reading about in the news lately. Due to the popularity of this letter on the Internet, a major nightly news program recently devoted an entire show to the investigation of the program described below to see if it really can make people money.
The show also investigated whether or not the program was legal. Their findings proved once and for all that there are absolutely no laws prohibiting the participation in the program. This has helped to show people that this is a simple, harmless and fun way to make some extra money at home.
The results of this show have been truly remarkable. So many people are participating that those involved are doing much better than ever before. Since everyone makes more as more people try it out, its been very exciting to be a part of lately. You will understand once you experience it.
HERE IT IS BELOW:
*** Print This Now For Future Reference ***
The following income opportunity is one you may be interested in taking a look at. It can be started with VERY LITTLE investment and the income return is TREMENDOUS!!!
$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$ If you would like to make at least $50,000 in less than 90 days ! Please read the enclosed program...THEN READ IT AGAIN!!! $$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$$
THIS IS A LEGITIMATE, LEGAL, MONEY MAKING OPPORTUNITY.It does not require you to come into contact with people, do any hard work, and best of all, you never have to leave the house except to get the mail. If you believe that someday you'll get that big break that you 'vebeen waiting for, THIS IS IT! Simply follow the instructions, andyour dreams will come true. This multi-level e-mail order marketingprogram works perfectly...100% EVERY TIME.
E-mail is the sales tool of the future. Take advantage of this non-commercialized method of advertising NOW!!! The longer you wait, the more people will be doing business using e-mail. Get your piece of this action!!!
MULTI-LEVEL MARKETING (MLM) has finally gained respectability. It is being taught in the Harvard Business School, and both Stanford Research and the Wall Street Journal have stated that between 50% and 65% of all goods and services will be sold through multi-level methods by the mid to late 1990's. This is a Multi-Billion Dollar industry and of the 500,000 millionaires in the U.S., 20% (100,000) made their fortune in the last several years in MLM. Moreover, statistics show 45 people become millionaires everyday through Multi-Level Marketing.
You may have heard this story before, but over the summer Donald Trump made an appearance on the David Letterman show. Dave asked him what he would do if he lost everything and had to start over from scratch. Without hesitating, Trump said he would find a good network marketing company and get to work. The audience started to hoot and boo him. He looked out at the audience and dead-panned his response: "That's why I'm sitting up here and you are all sitting out there!"
The enclosed information is something I almost let slip through my fingers. Fortunately, sometime later I re-read everything and gave somethought and study to it. My name is Johnathon Rourke. Two years ago, the corporation I worked at for the past twelve years down-sized and my position was eliminated. After unproductive job interviews, I decided to open my own business. Over the past year, I incurred many unforeseen financial problems. I owed my family, friends and creditors over $35,000. The economy was taking a toll on my business and I just couldn't seem to make ends meet. I had to refinance and borrow against my home to support my family and struggling business. AT THAT MOMENT something significant happened in my life and I am writing to share the experience in hopes that this will change your life FOREVER FINANCIALLY!!!
In mid December, I received this program via e-mail. Six month's prior to receiving this program I had been sending away for information on various business opportunities. All of the programs I received, in my opinion, were not cost effective. They were either too difficult for me to comprehend or the initial investment was too much for me to risk to see if they would work or not. One claimed that I would make a million dollars in one year...it didn't tell me I'd have to write a book to make it!
But like I was saying, in December of 1997 I received this program. I didn't send for it, or ask for it, they just got my name off a mailing list.THANK GOODNESS FOR THAT!!! After reading it several times, to make sure I was reading it correctly, I couldn't believe my eyes. Here was a MONEY MAKING PHENOMENON. I could invest as much as I wanted to start, without putting me further into debt. After I got a pencil and paper and figured it out, I would at least get my money back. But like most of you I was still a little skeptical and a little worried about the legal aspects of it all. So I checked it out with the U.S. Post Office (1-800-725-2161 24-hrs) and they confirmed that it is indeed legal! After determining the program was LEGAL and NOT A CHAIN LETTER, I decided "WHY NOT."
Initially I sent out 10,000 e-mails. It cost me about $15 for my time on-line. The great thing about e-mail is that I don't need any money for printing to send out the program, and because all of my orders are fulfilled via e-mail, my only expense is my time. I am telling you like it is I hope it doesn't turn you off, but I promised myself that I would not "rip-off" anyone, no matter how much money it made me.
In less than one week, I was starting to receive orders for REPORT #1 By January 13, I had received 26 orders for REPORT #1. Your goal is to "RECEIVE at least 20 ORDERS FOR REPORT #1 WITHIN 2 WEEKS. IF YOU DON'T, SEND OUT MORE PROGRAMS UNTIL YOU DO!" My first step in making $50,000 in 90 days was done. By January 30, I had received 196 orders for REPORT #2. Your goal is to "RECEIVE AT LEAST 100+ ORDERS FOR REPORT #2 WITHIN 2 WEEKS. IF NOT, SEND OUT MORE PROGRAMS UNTIL YOU DO. ONCE YOU HAVE 100 ORDERS, THE REST IS EASY, RELAX, YOU WILL MAKE YOUR $50,000 GOAL." Well, I had 196 orders for REPORT #2, 96 more than I needed. So I sat back and relaxed. By March 1, of my e-mailing of 10,000, I received $58,000 with more coming in every day.
I paid off ALL my debts and bought a much needed new car. Please take time to read the attached program, IT WILL CHANGE YOUR LIFE FOREVER!! ! Remember, it won't work if you don't try it. This program does work , but you must follow it EXACTLY! Especially the rules of not trying to place your name in a different place. It won't work and you'll lose out on a lot of money! In order for this program to work, you must meet your goal of 20+ orders for REPORT #1, and 100+ orders for REPORT #2 and you will make $50,000 or more in 90 days. I AM LIVING PROOF THAT IT WORKS!!!
If you choose not to participate in this program, I am sorry. It really is a great opportunity with little cost or risk to you. If you choose to participate, follow the program and you will be on your way to financial security. If you are a fellow business owner and are in financial trouble like I was, or you want to start your own business, consider this a sign. I DID!
Sincerely, Johnathon Rourke
A PERSONAL NOTE FROM THE ORIGINATOR OF THIS PROGRAM:
By the time you have read the enclosed program and reports, you should have concluded that such a program, and one that is legal, could not have been created by an amateur.
Let me tell you a little about myself. I had a profitable business for 10 years. Then in 1979 my business began falling off. I was doing the same things that were previously successful for me, but it wasn't working. Finally, I figured it out. It wasn't me, it was the economy. Inflation and recession had replaced the stable economy that had been with us since 1945.I don't have to tell you what happened to the unemployment rate... because many of you know from first hand experience. There were more failures and bankruptcies than ever before.
The middle class was vanishing. Those who knew what they were doing invested wisely and moved up. Those who did not, including those who never had anything to save or invest, were moving down into the ranks of the poor. As the saying goes, "THE RICH GET RICHER AND THE POOR GET POORER." The traditional methods of making money will never allow you to "move up" or "get rich", inflation will see to that.
You have just received information that can give you financial freedom for the rest of your life, with "NO RISK" and "JUST A LITTLE BIT OF EFFORT." You can make more money in the next few months than you have ever imagined. I should also point out that I will not see a penny of this money, nor anyone else who has provided a testimonial for this program. I have already made over 4 MILLION DOLLARS!I have retired from the program after sending thousands and thousands of programs.
Follow the program EXACTLY AS INSTRUCTED. Do not change it in any way . It works exceedingly well as it is now. Remember to e-mail a copy of this exciting report to everyone you can think of. One of the people you send this to may send out 50,000...and your name will be on everyone of them!
Remember though, the more you send out the more potential customers you will reach.
So my friend, I have given you the ideas, information, materials and opportunity to become financially independent. IT IS UP TO YOU NOW!
"THINK ABOUT IT"
Before you delete this program from your mailbox, as I almost did, take a little time to read it and REALLY THINK ABOUT IT. Get a pencil and figure out what could happen when YOU participate. Figure out the worst possible response and no matter how you calculate it, you will still make a lot of money! You will definitely get back what you invested. Any doubts you have will vanish when your first orders come in. IT WORKS!
Jody Jacobs, Richmond, VA
HERE'S HOW THIS AMAZING PROGRAM WILL MAKE YOU THOUSANDS OF DOLLAR$
INSTRUCTIONS:
This method of raising capital REALLY WORKS 100% EVERY TIME. I am sure that you could use up to $50,000 or more in the next 90 days. Before you say "BULL... ", please read this program carefully.
This is not a chain letter, but a perfectly legal money making opportunity. Basically, this is what you do: As with all multi-level businesses, we build our business by recruiting new partners and selling our products. Every state in the USA allows you to recruit new multi-level business partners, and we offer a product for EVERY dollar sent. YOUR ORDERS COME BY MAIL AND ARE FILLED BY E-MAIL, so you are not involved in personal selling. You do it privately in your own home, store or office. This is the GREATEST Multi-Level Mail Order Marketing anywhere.
This is what you MUST do:
1. Order all 4 reports shown on the list below (you can't sell them if youdon't order them). -- For each report, send $5.00 CASH, the NAME & NUMBER OF THE REPORT YOU ARE ORDERING, YOUR E-MAIL ADDRESS, and YOUR NAME & RETURN ADDRESS (in case of a problem) to the person whose name appears on the list next to the report. MAKE SURE YOUR RETURN ADDRESS IS ON YOUR ENVELOPE IN CASE OF ANY MAIL PROBLEMS! -- When you place your order, make sure you order each of the four reports. You will need all four reports so that you can save them on your computer and resell them. -- Within a few days you will receive, via e-mail, each of the four reports. Save them on your computer so they will be accessible for you to send to the 1,000's of people who will order them from you.
2. IMPORTANT DO NOT alter the names of the people who are listed next to each report, or their sequence on the list, in any way other than is instructed below in steps "a" through "f" or you will lose out on the majority of your profits. Once you understand the way this works, you'll also see how it doesn't work if you change it. Remember, this method has been tested,and if you alter it, it will not work. a. Look below for the listing of available reports. b. After you've ordered the four reports, take this advertisement and remove the name and address under REPORT #4. This person has made it through the cycle and is no doubt counting their $50,000! c. Move the name and address under REPORT #3 down to REPORT #4. d. Move the name and address under REPORT #2 down to REPORT #3. e. Move the name and address under REPORT #1 down to REPORT #2. f. Insert your name/address in the REPORT #1 position.
Please make sure you COPY ALL INFORMATION, every name and address, ACCURATELY!
3. Take this entire letter, including the modified list of names, and save it to your computer. Make NO changes to the instruction portion of this letter.
Your cost to participate in this is practically nothing (surely you can afford $20). You obviously already have an Internet connection and e-mail is FREE!
There are two primary methods of building your downline:
METHOD #1: SENDING BULK E-MAIL
Let's say that you decide to start small, just to see how it goes, and we'll assume you and all those involved send out only 2,000 programs each. Let's also assume that the mailing receives a 0.5% response. Using a good list the response could be much better. Also, many people will send out hundreds of thousands of programs instead of 2,000. But continuing with this example, you send out only 2,000 programs. With a 0.5% response, that is only 10 orders for REPORT #1. Those 10 people respond by sending out 2,000 programs each for a total of 20,000. Out of those 0.5%, 100 people respond and order REPORT #2. Those 100 mail out 2,000 programs each for a total of 200,000. The 0.5% response to that is 1,000 orders for REPORT #3. Those 1,000 send out 2,000 programs each for a 2,000,000 total. The 0.5% response to that is 10,000 orders for REPORT #4. That's 10,000 $5 bills for you. CASH!!! Your total income in this example is $50 + $500 + $5,000 + $50,000 for a total of $55,550!!! REMEMBER FRIEND, THIS IS ASSUMING 1,990 OUT OF THE 2,000 PEOPLE YOU MAIL TO WILL DO ABSOLUTELY NOTHING AND TRASH THIS PROGRAM! DARE TO THINK FOR A MOMENT WHAT WOULD HAPPEN IF EVERYONE, OR HALF SENT OUT 100,000 PROGRAMS INSTEAD OF 2,000. Believe me, many people will do justthat, and more! By the way, your cost to participate in this is practically nothing. You obviously already have an Internet connection and e-mail is FREE!!! REPORT #2 will show you the best methods for bulk e-mailing, tell you where to obtain free bulk e-mail software and where to obtain e-mail lists.
METHOD #2 - PLACING FREE ADS ON THE INTERNET
Advertising on the internet is very, very inexpensive, and there are HUNDREDS of FREE places to advertise. Let's say you decide to start small just to see how well it works. Assume your goal is to get ONLY 10 people to participate on your first level. (Placing a lot of FREE ads on the Internet will EASILY get a larger response.) Also assume that everyone else in YOUR ORGANIZATION gets ONLY 10 downline members. Follow this example to achieve the STAGGERING results below:
1st level--your 10 members with $5.......................................$50 2nd level--10 members from those 10 ($5 x 100)..................$500 3rd level--10 members from those 100 ($5 x 1,000)...........$5,000 4th level--10 members from those 1,000 ($5 x 10,000).....$50,000 THIS TOTALS ----------} $55,550
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///////////////////////////////////////////////////////////////// Remove at rhobbs12-at-yahoo.com /////////////////////////////////////////////////////////////////
-} We have a Zeiss TEM. Normally when the filament is first switched on with } both current and heating knobs are turned to mininimum the voltage indicator } shoots up to 2V then immeadiatly starts going down to 1.3-1.4V within no more } than 2 minutes, then we start raising the voltage by turning the heating knob } until the voltage meter reads between 1.5-1.6V (the recomended voltage stated in } the manual). } Recently though we have been noticing that the voltage shoots up to 2.2V and } stays there for several minutes then starts coming down very slowly and settles } at more than 1.6V!!... Needless to say that we have also been burning too many } filaments too quickly!! The filament lives have been no more than 10-15hours. } We would appreciate any hints or suggestions on why would this happen. We } are suspecting an electric problem in the filament control circuitry. Thank } you.
I have seen this happen in other filaments and I and a couple of EE's are at a loss to explain it. The solution we came up wiht was to use a current controlled power supply and slowly bring up the current insted of the voltage. You may have to switch power supplies when the heater is up to temperature.
Good luck Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
----------------------------------------------------. } } } Mid-$30K? Is this for real? Is this the going rate for SEM } specialists? That is about 1/4 of what I would consider. } There must be some subtle differentiation of what is expected } from such positions vs. the qualifications and experience of } others in the field. Am I the only one shocked about this? }
We have graduates with experience in electrical engineering technology graduating for 50k & 60 K my sun is still fishing he is probably going to take a lower paid job to get into embed systems but has passed up 70k as a system admin. He does have 10 years experience in the field. All but the 50K kid are older and have some real work behind them. Last I heard the 60K was the top at the university. It annoys some of the engineers that look at EET as where you go when you can't cut engineering.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
Sres MSA: Dado que mi tema de investigaci=F3n es el an=E1lisis de propiedades mec=E1nicas de pol=EDmeros semicritalinos, necesitar=EDa contactarme con alguna persona que pueda informarme sobre estudios de microestructura de los mismos.Desde ya muchas gracias.=20 Ing elena Brandaleze.
Judging from last summer's C&E News salary survey (7/27/98) $35k is about right for a new BS chemist. The average salary for 10 years experience goes up to $48k. The MS person that they prefer will cost substantially more however - $45-55k.
Now a second question is whether they'll be happy with a new BS in this position - I think not, but as in many things you do get what you pay for.
Richard Shalvoy Arch Chemicals (formerly Olin Corporation) Cheshire, CT
-----Original Message----- } From: Dr. Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Thursday, May 13, 1999 10:59 PM To: MSA listserver
Keep in mind that this is a university doing the hiring in a field where (at least in biological EM positions) people tend to enjoy their work. Also, my first job at a university only had a 7.5 hour workday. I agree that it's disappointing, but they figure someone with minimal graduate work/experience receiving full medical, tuition reimbursement, 4 weeks of vacation, and the casual atmosphere of a university, they can get away with it. Especially for someone with minimal prior experience.
I'm not trying to justify it, just trying to offer a window into how they are thinking. They will likely fill the position with some willing individual looking for that extra job experience and generous vacation time.
Gregg Sobocinski Parke-Davis Pharmaceuticals Ann Arbor, Michigan USA Gregg.Sobocinski-at-wl.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
} ********************************************************** } Job Announcement: } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } UNL Center for Materials Research and Analysis } } Analyze/characterize materials using electron microscopy, materials } preparation and computer instrumentation. Supervise/train students, } faculty and visiting researchers utilizing these methods. Bachelor's with } major in physical science, engineering or related field plus three years } experience in the operation, repair or design of electron microscopes or } other scientific instrumentation. Master's preferred. Must have excellent } computer and interpersonal/communication skills. TEM, SEM, x-ray } diffraction or materials sample preparation experience preferred. Excellent } benefits. Submit cover letter, resume and names, addresses and telephone } numbers of three professional references to Professor Brian Robertson, } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } will remain open until a suitable candidate is found. UNL is committed to } AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886. }
We are to begin the process of using tripod polishing for preparing TEM samples, and request vendors to contact me with regards to types of polishers available, style, cost, ease of use.
I also welcome users of this method to comment online (or email directly) as to the ease of use and technique of sample preparation, since our lab has not gone this route before.
thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
This is similar to what I make and I do SEM, TEM, LM and immuno. at LM and TEM levels for an animal research lab at UTSW in Dallas. I have over 15 years of experience, a MS, and the position I hold is considered a non-tenure, PhD position. This is reality for working in academia in the midwest (I should mention I started out at the U of MN, where I was paid alot less.). Why do I stay? Cost of living here is low, so you can live on it and I like the field so much I'm getting a PhD in neuroscience to continue to grow in it.
Karen Pawlowski Sr. Research Assoc. UT Southwestern, Dallas, TX PhD candidate, UT Dallas, Dallas, TX
On Thu, 13 May 1999, Dr. Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Mid-$30K? Is this for real? Is this the going rate for SEM } specialists? That is about 1/4 of what I would consider. } There must be some subtle differentiation of what is expected } from such positions vs. the qualifications and experience of } others in the field. Am I the only one shocked about this? } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } Dear colleagues, } } We want to hire a suitable person for the Central Facility for Electron } } Microscopy at the Center for Materials Research and Analysis at the } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } the range of the mid $30k's and is dependent on experience. The medical, } } dental and retirement benefits package is substantial and includes } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } growing, has low unemployment, is great for families, has a good range of } } live music, dance and theater, and has very good public and other schools.) } } The Facility provides user-access for ~60 faculty plus their students and } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } specimen preparation equipment, computers and darkroom. Some development } } of new research instrumentation is in progress for mapping of magnetic } } materials and more development is planned for the Facility. You can find } } out more about the Facility on our web site (that does need a little work) } } at URL: http://www.unl.edu/CMRAcfem/ } } Thanks for passing on the word, } } Brian Robertson } } } } ********************************************************** } } Job Announcement: } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } UNL Center for Materials Research and Analysis } } } } Analyze/characterize materials using electron microscopy, materials } } preparation and computer instrumentation. Supervise/train students, } } faculty and visiting researchers utilizing these methods. Bachelor's with } } major in physical science, engineering or related field plus three years } } experience in the operation, repair or design of electron microscopes or } } other scientific instrumentation. Master's preferred. Must have excellent } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } diffraction or materials sample preparation experience preferred. Excellent } } benefits. Submit cover letter, resume and names, addresses and telephone } } numbers of three professional references to Professor Brian Robertson, } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } will remain open until a suitable candidate is found. UNL is committed to } } AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886. } } } } } } *********************************************************** } } Assoc. Prof. Brian W. Robertson } } Department of Mechanical Engineering } } and Center for Materials Research and Analysis } } University of Nebraska-Lincoln, N124 WSEC, } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } Cheers, } Gary Gaugler, Ph.D. } } }
Sres MSA: Dado que mi tema de investigaci=F3n es el an=E1lisis de propiedades mec=E1nicas de pol=EDmeros semicritalinos, necesitar=EDa contactarme con alguna persona que pueda informarme sobre estudios de microestructura de los mismos.Desde ya muchas gracias.=20 Ing elena Brandaleze.=20
Elena Brandaleze asks whether there is anyone who can provide information on studies of the microstructure of semicrystalline polymers. She works on the mechanical properties of the same.
At 05:50 AM 5/14/99 , you wrote: } Gary: Where are all the 120K jobs. I might want one. } }
They are in Silicon Valley and all over California. These are senior positions. Junior and journey positions gross about $65K-$90K, plus benefits. A brand new person might start at $50K.
Nebraska and academia. Yep. Now I understand the low amount. Perhaps based on cost of living, it is not so bad. Cost of living here in CA is certainly higher. So hopefully, the compensation is relative and commensurate.
Just for your info, we have one of the best junior colleges in the country (and one of the few) that train and produce SEM techs. San Joaquin Delta College, Stockton CA has an excellent program and a fine reputation. With tons of biotech, microelectronics and materials companies all over the place, there are ample opportunities for people to get a start and to move up. Most companies would like people to have a BS but they really want someone who can do the job. They need results, not credentials.
Judy Murphy, who leads the Microscopy Technology Center at San Joaquin Delta College is on this list-server so if anyone has any questions about their program, I'm sure she would be glad to answer them.
My recommendation is to get someone to help you set up your lab and that can come in, spend a couple of days, and show you how to do it. I taught myself from the literature and it was a painful process. Once I could do it, then it was easy for me to show others how to do it. You should also get the literature from the MRS series of TEM sample prep books on how to do it and pay particular attention to Ron Anderson group's works. Number three has a fairly detailed outline of how to do it by John Benedict, Ron Anderson and Stanley Klepeis, MRS Vol 254, 121.
You should seriously consider the South Bay Technology course that is periodically taught. SBT has a very good relationship with Ron Anderson and since his group developed and taught the world the technique, I highly recommend that you contact them.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Fred Pearson To: microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hello
We are to begin the process of using tripod polishing for preparing TEM samples, and request vendors to contact me with regards to types of polishers available, style, cost, ease of use.
I also welcome users of this method to comment online (or email directly) as to the ease of use and technique of sample preparation, since our lab has not gone this route before.
thanks in advance
Fred
******************************************************** Fred Pearson Brockhouse Institute for Materials Research McMaster University 1280 Main St. West Hamilton, Ontario Canada L8S 4M1
This can end up being a very lengthy discussion for the listserver and I don't know your level of expertise with ultramicrotomy so I will keep this very brief. Please feel free to contact me or our applications staff directly for more detail.
You will need to be concerned with the Tg of each phase, their size and relative volumes in the sample. If the ABS phase is 80% and the other phases are not too different in Tg then start at 10-15 degrees above the ABS Tg.
If possible trim before putting in the Ultramicrotome, section at 100nm first to see how it behaves, keep the block face small (less than 1mm). You can try large faces later.
You are looking for consistent sections (no skips), no curling, no chip or flakes. You should see highly specular finish and relatively flat sections. There are a host of remedies for each variation, I will not try to put them all here. If you can get to 70nm you should have a good SPM sample.
Curling or skipping usually means sectioning too warm or a dull knife. Chips or flakes, with or instead of sections means one or more phases are too cold (glassy). Adjust temperature only a few degrees at a time.
A point that some people miss is that the back side of the sections are usually smoother than the front ( the block face which becomes the next section face). If the best you are able to do still shows some chatter try to hold a section face down and do your SPM on the back side.
We are a commercial manufacturer of Ultramicrotomes and cryo attachments to fit nearly all ultramicrotomes. We run a Materials Science Ultramicrotomy Course each Fall which addresses SPM sample preparation. Please see our web site for details.
www.Ventanamed.com Look for RMC button Steve Miller Director of North American Sales Ventana EM Products Group Ventana Medical Systems, Inc. 3450 S. Broadmont Tucson, AZ 85713 Direct phone: 520-205-4118 Fax: 520-903-0132
-----Original Message----- } From: Robert Plano [mailto:RPLANO-at-cea.com] Sent: Wednesday, May 12, 1999 3:51 PM To: 'spm-at-di.com'; 'Microscopy-at-Sparc5.Microscopy.Com'
Greetings.
I am working on making AFM topography and phase images of a series of polymer samples which are made up of varying amounts of polycarbonate, polypropylene and mostly (} 80%) ABS. I have some nice images already (soon to be on our web site) but want to improve the quality. I also want to look at the bulk regions by using cryo-microtomy to create a nice flat face for imaging. Can someone share their experiences with these types of samples and any tips on the microtomy technique (best temperatures, etc.)? I also plan to use plasma etching to see if that will clean up the surface.
Thanks in advance.
Robert J. Plano Staff Analyst, SPM Services Surface Science Labs (650)962-8767, ext. 742
You are not the only one shocked about this. Looking at the figure as low as Mid-$30K, I would say there is something seriously not quite right. These days a simple graduate in computer programming with 2-3 years experience easily gets a starting salary of $50k+ and as they go ahead with few more years of experience, they quickly move into the scale of $70k-80k and crosses $100k within a span of well less than 10 years of career.
- Is there something going wrong with microscopists and SEM/TEM engineers? - Is Mid-$30k fair to expect for electron microscopists (3 years experienced) who maintains the equipment as well? - What if they have 12 -15 years of experience? - Is $60k max (typically at Universities) is fair that these gorgeous people are expected to get max. in their lifetime? - Is that what the people who are the backbone for running and maintaining research facilities are supposed to rewarded with?
I mean just compare it with professionals with same qualifications and experience in other fields. In my personal opinion, there is something not quite right and needs to be re-assessed.
Zia ur Rahman University of Central Florida, Orlando, Florida
At 07:59 PM 5/13/1999 -0700, you wrote:
} Mid-$30K? Is this for real? Is this the going rate for SEM } specialists? That is about 1/4 of what I would consider. } There must be some subtle differentiation of what is expected } from such positions vs. the qualifications and experience of } others in the field. Am I the only one shocked about this? } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } Dear colleagues, } } We want to hire a suitable person for the Central Facility for Electron } } Microscopy at the Center for Materials Research and Analysis at the } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } the range of the mid $30k's and is dependent on experience. The medical, } } dental and retirement benefits package is substantial and includes } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } growing, has low unemployment, is great for families, has a good range of } } live music, dance and theater, and has very good public and other schools.) } } The Facility provides user-access for ~60 faculty plus their students and } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } specimen preparation equipment, computers and darkroom. Some development } } of new research instrumentation is in progress for mapping of magnetic } } materials and more development is planned for the Facility. You can find } } out more about the Facility on our web site (that does need a little work) } } at URL: http://www.unl.edu/CMRAcfem/ } } Thanks for passing on the word, } } Brian Robertson } } } } ********************************************************** } } Job Announcement: } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } UNL Center for Materials Research and Analysis } } } } Analyze/characterize materials using electron microscopy, materials } } preparation and computer instrumentation. Supervise/train students, } } faculty and visiting researchers utilizing these methods. Bachelor's with } } major in physical science, engineering or related field plus three years } } experience in the operation, repair or design of electron microscopes or } } other scientific instrumentation. Master's preferred. Must have excellent } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } diffraction or materials sample preparation experience preferred. Excellent } } benefits. Submit cover letter, resume and names, addresses and telephone } } numbers of three professional references to Professor Brian Robertson, } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } will remain open until a suitable candidate is found. UNL is committed to } } AA/EEO and ADA/504. If you require accommodation, please call (402) 472-7886. } } } } } } *********************************************************** } } Assoc. Prof. Brian W. Robertson } } Department of Mechanical Engineering } } and Center for Materials Research and Analysis } } University of Nebraska-Lincoln, N124 WSEC, } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } Cheers, } Gary Gaugler, Ph.D. } } } }
} -} We have a Zeiss TEM. Normally when the filament is first switched on } with } } both current and heating knobs are turned to mininimum the voltage } indicator } } shoots up to 2V then immeadiatly starts going down to 1.3-1.4V within no } more } } than 2 minutes, then we start raising the voltage by turning the heating } knob } } until the voltage meter reads between 1.5-1.6V (the recomended voltage } stated in } } the manual). } } Recently though we have been noticing that the voltage shoots up to 2.2V } and } } stays there for several minutes then starts coming down very slowly and } settles } } at more than 1.6V!!... Needless to say that we have also been burning too } many } } filaments too quickly!! The filament lives have been no more than } 10-15hours. } } We would appreciate any hints or suggestions on why would this happen. } We } } are suspecting an electric problem in the filament control circuitry. } Thank } } you. } } Gordon Cougar wrote: } I have seen this happen in other filaments and I and a couple of EE's are } at a loss to explain it. The solution we came up wiht was to use a current } controlled power supply and slowly bring up the current insted of the } voltage. } You may have to switch power supplies when the heater is up to temperature. } } Good luck } Gordon
Gordon has a good point. Also recall a recent post that suggested the use of "good" and "bad" filaments. That is, once you have a batch of filaments that are good, don't use up all of them...save a couple. Then, get a new batch, use them, and decide if they too are good. Then, when a batch performs differently than the previous good ones, you can verify that the earlier good ones are still good or that there is a vacuum leak (the post was based on filament lifetime and vacuum quality).
So, have you changed filament suppliers or gotten a new batch of filaments recently?
The nature of filaments is that their resistance is lowest when cold. As they heat, their resistance increases. Thus, for a given applied voltage, the circuit current will be highest at first, then decrease as the filament heats up. In a circuit where the current limiter is the filament (most circuits are this way), the circuit is generally constant voltage, not constant current.
What you are seeing is counter to what I would think is normal for sure. The only filament-based reason for the voltage to shoot up is if the filament's resistance shot up. That seems hard to explain. However, that the steady state voltage is higher than before could indicate a batch of filaments with higher intrinsic resistance. Without seeing the circuit diagram of the filament supply section, it is hard to make a qualified recommendation on it or even to speculate. Since the filament is at high potential, there are many novel methods that manufacturers use to achieve this and to supply an isolated filament current.
You might also check that the filament is well seated, the terminals are tight and all associated connections are tight.
~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Modern surfers use PC boards. You can too at http://photoweb.net ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
OK OK can we move this "debate" off the list? It has been pointed out that academic salaries are low compared to industry (always have been, always will be), that people in certain fields (e.g., engineers - why not use MDs for a real eye-opener?!) make more than those from other fields (e.g., biologists) at all levels of experience, that salaries in any position fluctuate with location, and that academics are generally not satisfied with their salaries (once they discover how much others are making!). I think that covers all the bases.... OOPS I forgot the gender comparision - maybe someone can throw in one final thought-provoking message along those lines.
Rob Palmer Academic
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At 09:19 AM 5/14/99 , you wrote: } Welcome to our world!!! This is pretty much the going rate...sad, isn't } it? For biological microscopists, too. Techs don't get paid very well. } } Tamara Howard } CSHL }
Yeah...very sad and perplexing. Microscopy and EM is not at all a trivial undertaking. Especially if one is looking for useful results. TEM work is even more challenging from what I have seen. I am on the SEM side.
While I admit that I am not 100% a microscopist (it is a means to another end), I am awed by the pool of expertise out there. I don't claim to be an expert by any stretch and I appreciate the free flow of information on this list-server and the one-on-one communications.
I know that there are well-paying tech jobs out there--my brother had several. But even he jumped ship for software work. I suppose that pay is geographically based and contorted by supply and demand. I can also understand low pay in an academic instutution where non-academia is viewed differently. Nevertheless, expertise is not something that just happens. Maybe "You get what you pay for" applies here? In Northern CA, and especially Silicon Valley, EM techs make good wages. And they are in demand. Unfortunately, at both U.S. Coasts, the semiconductor business seems to be imploding. In this respect, I'm like a vulture, waiting for a great deal on a recent SEM being dumped by a company. But for jobs, the only bright spot appears to be biotech.
There is also a vocational institute in Madison Wisconsin that teaches electron microscopy. The graduates of this program are well trained in SEM, TEM, AFM, FIB, materials and biological prep, etc. For more information contact Glenn Boda at (608) 246-6254 or Michael Kostrna at (608) 246-6762.
A textbook editor is looking for a micrograph. He isn't a list subscriber, so please contact him directly.
} ...the image we need as given to us from the chapter editor: } } A picture of aluminum atoms from a scanning tunneling microscope. The } individual atoms of aluminum should appear as dots. The individual atoms } in } the photograph may be accentuated by adding color so that it is easier } to see } them. This image would be used in an upcoming high school Science } textbook.
} Andy Christiansen } Photo Researcher } Holt, Rinehart and Winston } achristiansen-at-hrw.com
Caroline Schooley Educational Outreach Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/PCI/pci.html
I know there have been some discussions on this listserver in the past about computer sign up systems for electron microscopes. Are there any new programs out there? We are working on a web-based instrument sign up program to replace our existing sign up/microscope accounting system. The old system is not Y2K friendly. Besides the Universty of Minnesota, is there any other group which has a working web-based system?
John C. Wheatley Lab Manager Arizona State University Center for Solid State Science PSA-213 BOX 871704 Tempe, AZ 85287-1704
Actually this salary range is not as low as you would think.. I have a M.S. degree and 5 years of EM experience and it took a while for me to find a job in EM throughout the country.. I was applying for jobs that were offering $25K to start as a salary and some were more depending on what part of the country you were looking in... I ended up in Seattle at a job for $32K with benefits to start... I am now currently in Los Angeles at UCLA Medical Center and the salary here is quite a bit more than in Seattle.. But that is outweighed due to the high cost of just living here...So the salary for this job is not as bad as one might think...
==========================
} You are not the only one shocked about this. Looking at the figure as low } as Mid-$30K, I would say there is something seriously not quite right. } These days a simple graduate in computer programming with 2-3 years } experience easily gets a starting salary of $50k+ and as they go ahead with } few more years of experience, they quickly move into the scale of $70k-80k } and crosses $100k within a span of well less than 10 years of career. } } - Is there something going wrong with microscopists and SEM/TEM engineers? } - Is Mid-$30k fair to expect for electron microscopists (3 years } experienced) who maintains the equipment as well? } - What if they have 12 -15 years of experience? } - Is $60k max (typically at Universities) is fair that these gorgeous } people are expected to get max. in their lifetime? } - Is that what the people who are the backbone for running and maintaining } research facilities are supposed to rewarded with? } } I mean just compare it with professionals with same qualifications and } experience in other fields. In my personal opinion, there is something not } quite right and needs to be re-assessed. } } Zia ur Rahman } University of Central Florida, } Orlando, Florida } } } At 07:59 PM 5/13/1999 -0700, you wrote: } } } Mid-$30K? Is this for real? Is this the going rate for SEM } } specialists? That is about 1/4 of what I would consider. } } There must be some subtle differentiation of what is expected } } from such positions vs. the qualifications and experience of } } others in the field. Am I the only one shocked about this? } } } } } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } } } Dear colleagues, } } } We want to hire a suitable person for the Central Facility for Electron } } } Microscopy at the Center for Materials Research and Analysis at the } } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } } the range of the mid $30k's and is dependent on experience. The medical, } } } dental and retirement benefits package is substantial and includes } } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } } growing, has low unemployment, is great for families, has a good range of } } } live music, dance and theater, and has very good public and other schools.) } } } The Facility provides user-access for ~60 faculty plus their students and } } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } } specimen preparation equipment, computers and darkroom. Some development } } } of new research instrumentation is in progress for mapping of magnetic } } } materials and more development is planned for the Facility. You can find } } } out more about the Facility on our web site (that does need a little work) } } } at URL: http://www.unl.edu/CMRAcfem/ } } } Thanks for passing on the word, } } } Brian Robertson } } } } } } ********************************************************** } } } Job Announcement: } } } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } } } UNL Center for Materials Research and Analysis } } } } } } Analyze/characterize materials using electron microscopy, materials } } } preparation and computer instrumentation. Supervise/train students, } } } faculty and visiting researchers utilizing these methods. Bachelor's with } } } major in physical science, engineering or related field plus three years } } } experience in the operation, repair or design of electron microscopes or } } } other scientific instrumentation. Master's preferred. Must have excellent } } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } } diffraction or materials sample preparation experience preferred. Excellent } } } benefits. Submit cover letter, resume and names, addresses and telephone } } } numbers of three professional references to Professor Brian Robertson, } } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } } will remain open until a suitable candidate is found. UNL is committed to } } } AA/EEO and ADA/504. If you require accommodation, please call (402) } 472-7886. } } } } } } } } } *********************************************************** } } } Assoc. Prof. Brian W. Robertson } } } Department of Mechanical Engineering } } } and Center for Materials Research and Analysis } } } University of Nebraska-Lincoln, N124 WSEC, } } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } } } Cheers, } } Gary Gaugler, Ph.D. } } } } } } } } }
We have a researcher who would like to use the technique of Green and Linstead (Protoplasma (1990) 158:33-38) for making a mold, then a cast of plant shoot apices. They reference dental impression polymers from Kerr UK Ltd., Peterborough, UK. I may have missed it but I didn't find it (or the likes) in the half-dozen mainstream microscopy suppliers' catalogs we have on hand.
Can anyone suggest a source for these products? It would probably be easiest and fastest if we could get a supplier in the USA.
Thanks in advance for any help you can give.
+++++++++++++++++ Dale A. Callaham Senior Microscopist, Stuck Jar Lid Opener, Odd Job Bear Central Microscopy Facility The University of Massachusetts Amherst, MA 01007 ------------------------- Phone 413-545-3751 FAX 413-545-3243 email dac-at-bio.umass.edu
I am looking for an objective aperture for a Hitachi H-9000 (UHV) TEM with sizes of 10,20,30 & 50 microns. Any suggestions about suppliers -- we can supply dimensions.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
Hi, The subject line says all: I am using LM to look at sections embedded with Spurr's, and I would like to know its refractive index. Haven't been able to track it down on the web or through my vendor.
Thanks, Jill
------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
Hi, I would like to initiate a discussion with someone about the following specific questions, but I don't want to bog down the list. If you can help, please reply directly to me.
First of all, I'm not much of a microscopist and becoming more experienced in order to work on a particular problem: where are bacteria located on undisturbed sand grains. I embed "undisturbed" samples with Spurr's (after staining), cut (very) thick sections (0.5 mm) and look at them with fluorescence microscopy (100 x oil objective).
I would like to look at the sections without using cover slips for the following reasons:
1) Reduced sample preparation time if I am looking "into" the sample rather than right at the surface. I don't have to be as careful in preparing the surface, and can worry less about the polishing compounds becoming embedded in the resin.
2) I have actually found better results ("better images" to my eye) looking into the sample, and this confuses me. The resin itself is autofluorescent, and the DAPI-stained bacteria are brighter still. Could it be that by placing resin between the "focal plane" (quotes because I'm not sure if it's the correct term) and the objective that I am attenuating some of the light with the resin, and therefore can see the contrast between the bacteria and the resin better? I have found that stopping down the diaphragm near the light source helps too.
Now, I know that if the resin does not have the same refractive index as glass+oil, it will lead to distortion. My question is, does it matter for my purposes: I am _not_ looking at the size of the bacteria, just presence/absence. I am tracing the sediment-grain edges (I have a digital camera hooked up to the microscope). Wouldn't the distortion merely affect the apparent width of the grain broundary, rather than the shape (pattern) of the grain outline itself? The grains are 50-200 micrometer in diameter, i.e. a whole grain will not typically be within a field of view.
I am mainly after a record of where the bacteria are located on grain edges, and am not in need of an exact visual representation. I am would rather have large sample sizes (thus the desire for reduced prep time) rather than high precision because I want to be able to treat the results (bacterial position compared with pore width) statistically. I want to be able to publish these results, and don't want to proceed ahead with what just "works best" if it doesn't make any sense.
In searching the archives, I came across someone's lament about the lack of curricula for microscopy and I completely agree! I am at a major research university and there are no classes given on light microscopy.
One other question is: we have a 100x oil objective NA 1.25 (Zeiss) with a collar that adjusts from .8 - 1.25. The microscope owner _thinks_ this is to adjust for different refractive indexes of the specimen. I can see that it functions as an aperature, and because it goes up to 1.25 is seems that it would be related to NA. I can't understand why someone would want a lower NA, unless it would increase the "depth of field". Any ideas? Do you control NA and refractive index the same way?
Thanks for any help you can give me, Jill ------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
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Reply to: Re: Electron Microscopy Position Available = Immediately An interesting topic that I hope will be explored in a little more detail, = is not so much how microscopists are paid (which is usually too little) = but what is expected of them. In addition to knowing evey preparation = technique and piece of equipment, we have to know how to fix the equipment,= teach students and post-docs and figure out ways to fund the facility. = No problem.
The really dangerous aspect to this is that quite often the head of the EM = lab is expected to provide a data collection and image interpretation = service for the majority of users. Although I am sure everyone feels able = to do this, for microscopy (and in particular electron microscopy) to = continue to grow, it is important that individual researchers become = responsible for their own specimen preparation, data collection and = results interpretation. Not knowing the technology is no longer a good = excuse to avoid the work. =
On the subject of pay, I always thought that the low initial salaries = being offered were to cover the risk involved with new hires. The true = story could only be that once settled in the salary actually rose to a = comfortable level! =
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Karen S Pawlowski wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
} TEM levels for an animal research lab at UTSW in Dallas. I have over 15 } years of experience, a MS, and the position I hold is considered a } non-tenure, PhD position. This is reality for working in academia in the } midwest (I should mention I started out at the U of MN, where I was paid } alot less.). Why do I stay? Cost of living here is low, so you can live = } on it and I like the field so much I'm getting a PhD in neuroscience to } continue to grow in it. } } Karen Pawlowski } Sr. Research Assoc. UT Southwestern, Dallas, TX } PhD candidate, UT Dallas, Dallas, TX } } On Thu, 13 May 1999, Dr. Gary Gaugler wrote: } } } ------------------------------------------------------------------------=
} } The Microscopy ListServer -- Sponsor: The Microscopy Society of America = } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.= Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.= html } } -----------------------------------------------------------------------.=
} } = } } = } } Mid-$30K? Is this for real? Is this the going rate for SEM } } specialists? That is about 1/4 of what I would consider. } } There must be some subtle differentiation of what is expected } } from such positions vs. the qualifications and experience of = } } others in the field. Am I the only one shocked about this? } } = } } = } } = } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } } } Dear colleagues, } } } We want to hire a suitable person for the Central Facility for = Electron } } } Microscopy at the Center for Materials Research and Analysis at the } } } University of Nebraska-Lincoln (UNL). The salary will at least likely = be in } } } the range of the mid $30k's and is dependent on experience. The = medical, } } } dental and retirement benefits package is substantial and includes } } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } } growing, has low unemployment, is great for families, has a good range = of } } } live music, dance and theater, and has very good public and other = schools.) = } } } The Facility provides user-access for ~60 faculty plus their students = and } } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A = SEM } } } and a VG HB501 field-emission STEM, along with a full range of = accessories, } } } specimen preparation equipment, computers and darkroom. Some = development } } } of new research instrumentation is in progress for mapping of magnetic } } } materials and more development is planned for the Facility. You can = find } } } out more about the Facility on our web site (that does need a little = work) } } } at URL: http://www.unl.edu/CMRAcfem/ = } } } Thanks for passing on the word, } } } Brian Robertson } } } } } } ********************************************************** } } } Job Announcement: } } } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } } } UNL Center for Materials Research and Analysis } } } } } } Analyze/characterize materials using electron microscopy, materials } } } preparation and computer instrumentation. Supervise/train students, } } } faculty and visiting researchers utilizing these methods. Bachelor's = with } } } major in physical science, engineering or related field plus three = years } } } experience in the operation, repair or design of electron microscopes = or } } } other scientific instrumentation. Master's preferred. Must have = excellent } } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } } diffraction or materials sample preparation experience preferred. = Excellent } } } benefits. Submit cover letter, resume and names, addresses and = telephone } } } numbers of three professional references to Professor Brian Robertson, } } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-= 0656 or } } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Posit= ion } } } will remain open until a suitable candidate is found. UNL is = committed to } } } AA/EEO and ADA/504. If you require accommodation, please call (402) = } 472-7886. } } } } } } } } } *********************************************************** } } } Assoc. Prof. Brian W. Robertson } } } Department of Mechanical Engineering } } } and Center for Materials Research and Analysis } } } University of Nebraska-Lincoln, N124 WSEC, } } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } = } } Cheers, } } Gary Gaugler, Ph.D. } } = } } = } } = } } } } } RFC822 header } ----------------------------------- } } Received: from Sparc5.Microscopy.Com [206.69.208.10] by mailhouse.hei.= org } (SMTPD32-4.07) id A18E691011E; Fri, 14 May 1999 13:03:26 PST } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.= 11) = } id IAA02338 for dist-Microscopy; Fri, 14 May 1999 08:40:47 -0500 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by = } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id IAA02333 for = } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 14 May 1999 08:40:16 -= 0500 } Received: from poteidaia.utdallas.edu (poteidaia.utdallas.edu = } [129.110.10.1]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id = IAA02322 for = } {Microscopy-at-sparc5.microscopy.com} ; Fri, 14 May 1999 08:40:03 -0500 } Received: from apache.utdallas.edu (kna101-at-apache.utdallas.edu [129.110.= 16.9]) } by poteidaia.utdallas.edu (8.9.1/8.9.1/null-3.5) with ESMTP id IAA14724;=
} Fri, 14 May 1999 08:42:30 -0500 (CDT) } Received: from localhost (kna101-at-localhost) } by apache.utdallas.edu (8.9.1/8.9.1) with SMTP id IAA01748; } Fri, 14 May 1999 08:42:29 -0500 (CDT) } X-Authentication-Warning: apache.utdallas.edu: kna101 owned process = doing -bs } Date: Fri, 14 May 1999 08:42:29 -0500 (CDT) } From: Karen S Pawlowski {kna101-at-utdallas.edu} } To: "Dr. Gary Gaugler" {gary-at-gaugler.com} } cc: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Re: Electron Microscopy Position Available Immediately } In-Reply-To: {4.1.19990513195604.009dc490-at-pop.calweb.com} } Message-ID: {Pine.GSO.3.96.990514083332.1144A-100000-at-apache.utdallas.edu} =
Working together with process engineers and laser designers, we aim to improve the performance of current generation high power lasers and to develop the next.
If you are at, or near the end of a PhD thesis in microscopy (TEM), preferably in the semiconductor area, and are looking to widen your horizons in the world of industry, this could be for you.
A basic knowledge of german is essential.
If you are interested please contact our Human Resources Manager, Teresa Eichholzer, Binzstrasse 17, 8045 Zurich, Switzerland, or by e-mail at teresa.eichholzer-at-uniphasele.com
For further information please visit our website at http://www.uniphase.com.
-----Mensaje original-----De: Antonio Molina {ifrm111-at-if.csic.es} Para: Microscopy-at-MSA.Microscopy.Com {Microscopy-at-MSA.Microscopy.Com} Fecha: viernes 14 de mayo de 1999 17:06Asunto: LM, SEM (may be TEM or AFM) - Ice cristal size and shape: how to do it Dear microscopists, I am interested in measuring the size, shape, growth, dynamics of recrystallization, etc, of ice, contained in a variety of foods, and generated through different freezing processes (different freezing speeds, hydrostatic pressure, antifreezers). I wonder which could be the easier way to obtain quantitative data. I am considering light microscopy with a cryostage, but I don't know if I will be needing special optics (will phase contrast or polarisation help?). I am also starting to do experiments on cryo-SEM, but I am frightened of altering the ice distribution of my sample, if I cool it further. If you have come across with this problem, your comments could be very useful to me. Thanks, Antonio D. Molina-Garcia Inst. del Frio, CSIC 28040 Madrid SPAINPhone (+34) 91 5445607 (+34) 91 5492300 Fax (+34) 91 5493627E-mail : ifrm111-at-if.csic.es http://www.csic.es/ifrio/ingind.htm
} Date: Fri, 14 May 1999 10:57:51 +0000 } To: "Shalvoy, Richard B CHES" {RBShalvoy-at-archchemicals.com} } From: Lou Solebello {microls1297-at-mindspring.com} } Subject: RE: Electron Microscopy Position Available Immediately } X-Attachments: A:\loucv98.doc; } In-Reply-To: {39E38D236504D211B2BD00805FE616BF3DF9F6-at-ealt-exc-02.corp.olin.com} } } Thats about where I was in my last position. A similar, yet less comprehensive survey of microscopist salaries by Microscopy Today put the AVERAGE national salary for an MS degreed microscopist in all fields with 10+ yrs experience at $57,000 per year. One has to consider that this is an average, across all ecomomic demographics from a college par timer to a CEO. } } Would they be happy with a BS chemist out of school for $35k? probably not.....who is going to teach him, be his mentor? They are expecting too much for too little which makes an excellent recipe for disaster. The level of responsibility and knowledge they are expecting is way out of line for the compensation. } } I almost am embarrased at what I was earning in my last position, and I am considered to be "Senior level". I was earning $46,000/yr prior to benes which worked out to be about $60k/yr with benes. That however is my fault for not being savy enough business wise to research what I was worth. But, I didnt view it that way. Naive as it may sound, I too honesty in compensation for granted because I do not take advantage of others. Besides, I was doing what I love to do, which I cannot put a dollar amount on. A quick glance at my CV (attached) clearly shows that I am not average, but a good deal above average as a microanalyst. You might also be surprised to know that I earned less than that at well known prestigious microanalytical company where I had been for close to 10 yrs. That should dispel quite a few myths people have about the earnings of scientists that company....they are lower than you would expect. I have also worked with others making in the six digit range, and simply was amazed because they completely lacked the intelligence or expertise that would normally be warranted for that salary level. In fact, they shouldnt have been making a technicians salary. } } I thank you for humoring my dissertation on this subject, but I believe that everyone in the scientific community should be alarmed and discuss this topic mor openly. We wonder why there is scientific fraud, and many people have a negative opinion of scientists?, or why companies have been taking the attitudes they have? It is self promulgated by those same companies who are blind to their own errors, but have egos too big to admit it, and need to protect and justify their own high six digit figures. I personally do not like unions, but I strongly believe that if scientists do not do something about the situation, we will be our own demise by simply not acknowledging the problem. } } Lou Solebello } At 07:02 AM 5/14/99 -0500, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Not all jobs in California pay as much as Gary says. I'm at UCB and don't make anywhere near that much (and I've been doing this for 17 years). Maybe in Silicon Valley in the semiconductor business, but not in academia. I guess academic technicians are underpaid all over the states. Unfortunately, it's very expensive to live in to SF bay area unless you live east about an hour drive.
But we know what we're in for when we take the job. So I guess it's a case of let the technician beware.
Paula :-)
} At 05:50 AM 5/14/99 , you wrote: } } Gary: Where are all the 120K jobs. I might want one. } } } } } } They are in Silicon Valley and all over California. These } are senior positions. Junior and journey positions gross } about $65K-$90K, plus benefits. A brand new person might } start at $50K. }
Paula Sicurello UC Berkeley Electron Microscope Lab psic-at-uclink4.berkeley.edu phone: 510-642-2085 fax: 510-643-6207 http://biology.berkeley.edu/EML
Friends of mine with similar degrees and experience in other fields have second cars, vaction homes and boats. I have a 13 year old Honda. Of course, if she gets accepted, my daughter will go to BU for four years, tuition free 8-)
Ron ----- Original Message ----- } From: Robert J. Palmer Jr. {rjpalmer-at-utkux.utcc.utk.edu} ,Snip} - } maybe someone can throw in one final thought-provoking message along those } lines. } } Rob Palmer } Academic } } } Gordon, } } } } You are not the only one shocked about this. Looking at the figure as low } } as Mid-$30K, I would say there is something seriously not quite right. } } These days a simple graduate in computer programming with 2-3 years } } experience easily gets a starting salary of $50k+ {snip}
Henrik Kaker wrote: ============================================= I'm looking e-mail address of Graticules Limited from UK. ============================================== Try the following:
Mr. Brian Kyte General Manager Pyser Ltd./Graticules E-mail: BrianKyte-at-debscom.demon.co.uk
Yes, this is the same Brian Kyte formerly associated with VG Microtech and Polaron and Bio-Rad.
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: www.2spi.com ############################ =======================================================
I am editing some early papers of Sigmund Freud from the years before he turned to the mind. He published several papers in the 1870s and 1880s on his microscopical work, mainly on the nervous system of lower vertebrates.
He refers to some lenses, stains and fixation techniques I'm familiar with and some I'm not familiar with. For instance, he mentions Hartnac lenses of various numbers and serum iodide.
Does anyone know the state of microscopy during this period? Is there a person familiar with this era or book that might list the various stains, etc. in use? Does anyone know how to translate the Hartnac numbers into something meaningful to us? I suppose they refer to what we call power. Even general information such as what was visible and what not, whether artificial light or only sunlight was used, etc. would be helpful. He does not name the microscope he uses so we have to surmise what what available and standard.
Any help will be greatly appreciated and acknowledged.
- Larry Kunstadt, PhD The Psychoanalytic Institute New York University Medical Center
Kerr Specialties as a water soluble wax http://www.kerrcasting.com/Product/English/Wax/Sol-u-wax.htm It melts at 140 F and will dissolve in water. Would this have a use in embedding with out having to fully clear the subject. It will clear a lot faster after it is sliced than before. Just a thought.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
We are also considering developing a web-based reservation system, and I think many others would also be interested what other labs are doing in this area. I would be interested in learning of any software which is freely available or low cost and open source.
We are in the very early stages of modifying an open source (Perl) web-calender and scheduling system to suite our needs. The reference url is: http://curiosityshoppe.tierranet.com/framecal/index.shtml
I found this and a few others by searching the web with I believe was combinations of ("resource" and "scheduling" and "calenders")
If we develop a system based on FrameCal or something else we will make it freely available as licensing permits.
Of course, if someone else has a suitable system available freely or at low cost it would save us development time and costs.
} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.
} } I know there have been some discussions on this listserver in the past
} about computer sign up systems for electron microscopes. Are there any new } programs out there? We are working on a web-based instrument sign up } program to replace our existing sign up/microscope accounting system. The } old system is not Y2K friendly. Besides the Universty of Minnesota, is } there any other group which has a working web-based system? } } John C. Wheatley } Lab Manager } Arizona State University } Center for Solid State Science } PSA-213 } BOX 871704 } Tempe, AZ 85287-1704 } } Phone: (602) 965-3831 } FAX: (602) 965-9004 } John.Wheatley-at-ASU.Edu
At 01:22 PM 5/14/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Try Energy Beam Sciences
http://www.ebsciences.com
800.992.9037
They offer a wide assortment of apertures and also strip apertures for TEMs custom made to order. Not sure if they support Hitachi.
I would like to invite you to opening Image gallery of cartoons on www.coleoptera.org .
I am wonder if someone would like to put SEM nice picture of beetles to Image gallery.
Keep care and be of good cheer.
Regards
Ricardo
Only after the last tree has been cut down, only after the last river has been poisoned, only after the last fish has been caught, only then will you find that money can not be eaten.' CREE INDIAN PROPHECY.
Incoming mail is certified Virus Free. Checked by AVG anti-virus system (http://www.grisoft.com). Version: 6.0.37 / Virus Database: 30 - Release Date: 6.4.1999
L. D. Marks wrote: } } ------------------------------------------------------------------------} The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------.} } I am looking for an objective aperture for a } Hitachi H-9000 (UHV) TEM with sizes of 10,20,30 & 50 } microns. Any suggestions about suppliers -- we } can supply dimensions. } } ++++++++++++++++++++++++++++++++++++++++++++++++ } Laurence Marks } Department of Materials Science and Engineering } Northwestern University } fax: (847) 491-7820 } mailto:l-marks-at-nwu.edu } http://www.numis.nwu.edu } ++++++++++++++++++++++++++++++++++++++++++++++++
Dear Mr. Marks,
Ladd Research has been drilling apertures for EM use for the last forty plus years and we would be very happy to quote you on this. Please contact me directly and we can compare notes on sizes and give you a price. Since we drill ourselves we can give you a choice on hole sizes if you wish to vary from the norm.
Sincerely,
JD Arnott President
-- LADD RESEARCH 13 Dorset Lane Williston, VT 05495 TEL 1-800-451-3406 (US) or 1-802-878-6711 (anywhere) FAX 1-802-878-8074 e-mail ladres-at-worldnet.att.net web site http://www.ladd.cc
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Hi, I would like to initiate a discussion with someone about the following specific questions, but I don't want to bog down the list. If you can help, please reply directly to me.
First of all, I'm not much of a microscopist and becoming more experienced in order to work on a particular problem: where are bacteria located on undisturbed sand grains. I embed "undisturbed" samples with Spurr's (after staining), cut (very) thick sections (0.5 mm) and look at them with fluorescence microscopy (100 x oil objective).
I would like to look at the sections without using cover slips for the following reasons:
1) Reduced sample preparation time if I am looking "into" the sample rather than right at the surface. I don't have to be as careful in preparing the surface, and can worry less about the polishing compounds becoming embedded in the resin.
2) I have actually found better results ("better images" to my eye) looking into the sample, and this confuses me. The resin itself is autofluorescent, and the DAPI-stained bacteria are brighter still. Could it be that by placing resin between the "focal plane" (quotes because I'm not sure if it's the correct term) and the objective that I am attenuating some of the light with the resin, and therefore can see the contrast between the bacteria and the resin better? I have found that stopping down the diaphragm near the light source helps too.
Now, I know that if the resin does not have the same refractive index as glass+oil, it will lead to distortion. My question is, does it matter for my purposes: I am _not_ looking at the size of the bacteria, just presence/absence. I am tracing the sediment-grain edges (I have a digital camera hooked up to the microscope). Wouldn't the distortion merely affect the apparent width of the grain broundary, rather than the shape (pattern) of the grain outline itself? The grains are 50-200 micrometer in diameter, i.e. a whole grain will not typically be within a field of view.
I am mainly after a record of where the bacteria are located on grain edges, and am not in need of an exact visual representation. I am would rather have large sample sizes (thus the desire for reduced prep time) rather than high precision because I want to be able to treat the results (bacterial position compared with pore width) statistically. I want to be able to publish these results, and don't want to proceed ahead with what just "works best" if it doesn't make any sense.
In searching the archives, I came across someone's lament about the lack of curricula for microscopy and I completely agree! I am at a major research university and there are no classes given on light microscopy.
One other question is: we have a 100x oil objective NA 1.25 (Zeiss) with a collar that adjusts from .8 - 1.25. The microscope owner _thinks_ this is to adjust for different refractive indexes of the specimen. I can see that it functions as an aperature, and because it goes up to 1.25 is seems that it would be related to NA. I can't understand why someone would want a lower NA, unless it would increase the "depth of field". Any ideas? Do you control NA and refractive index the same way?
Thanks for any help you can give me, Jill ------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
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I am looking for an objective aperture for a Hitachi H-9000 (UHV) TEM with sizes of 10,20,30 & 50 microns. Any suggestions about suppliers -- we can supply dimensions.
++++++++++++++++++++++++++++++++++++++++++++++++ Laurence Marks Department of Materials Science and Engineering Northwestern University fax: (847) 491-7820 mailto:l-marks-at-nwu.edu http://www.numis.nwu.edu ++++++++++++++++++++++++++++++++++++++++++++++++
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Reply to: Re: Electron Microscopy Position Available Immediately An interesting topic that I hope will be explored in a little more detail, is not so much how microscopists are paid (which is usually too little) but what is expected of them. In addition to knowing evey preparation technique and piece of equipment, we have to know how to fix the equipment, teach students and post-docs and figure out ways to fund the facility. No problem.
The really dangerous aspect to this is that quite often the head of the EM lab is expected to provide a data collection and image interpretation service for the majority of users. Although I am sure everyone feels able to do this, for microscopy (and in particular electron microscopy) to continue to grow, it is important that individual researchers become responsible for their own specimen preparation, data collection and results interpretation. Not knowing the technology is no longer a good excuse to avoid the work. On the subject of pay, I always thought that the low initial salaries being offered were to cover the risk involved with new hires. The true story could only be that once settled in the salary actually rose to a comfortable level!
Paul Webster, Ph.D House Ear Institute 2100 West Third Street Los Angeles, CA 90057 phone:213 273 8026 fax: 213 413 6739 e-mail: pwebster-at-hei.org http://www.hei.org/htm/aemi.htm
Karen S Pawlowski wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dr. Gaugler, } } This is similar to what I make and I do SEM, TEM, LM and immuno. at LM and } TEM levels for an animal research lab at UTSW in Dallas. I have over 15 } years of experience, a MS, and the position I hold is considered a } non-tenure, PhD position. This is reality for working in academia in the } midwest (I should mention I started out at the U of MN, where I was paid } alot less.). Why do I stay? Cost of living here is low, so you can live } on it and I like the field so much I'm getting a PhD in neuroscience to } continue to grow in it. } } Karen Pawlowski } Sr. Research Assoc. UT Southwestern, Dallas, TX } PhD candidate, UT Dallas, Dallas, TX } } On Thu, 13 May 1999, Dr. Gary Gaugler wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Mid-$30K? Is this for real? Is this the going rate for SEM } } specialists? That is about 1/4 of what I would consider. } } There must be some subtle differentiation of what is expected } } from such positions vs. the qualifications and experience of } } others in the field. Am I the only one shocked about this? } } } } } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } } } Dear colleagues, } } } We want to hire a suitable person for the Central Facility for Electron } } } Microscopy at the Center for Materials Research and Analysis at the } } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } } the range of the mid $30k's and is dependent on experience. The medical, } } } dental and retirement benefits package is substantial and includes } } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } } growing, has low unemployment, is great for families, has a good range of } } } live music, dance and theater, and has very good public and other schools.) } } } The Facility provides user-access for ~60 faculty plus their students and } } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } } specimen preparation equipment, computers and darkroom. Some development } } } of new research instrumentation is in progress for mapping of magnetic } } } materials and more development is planned for the Facility. You can find } } } out more about the Facility on our web site (that does need a little work) } } } at URL: http://www.unl.edu/CMRAcfem/ } } } Thanks for passing on the word, } } } Brian Robertson } } } } } } ********************************************************** } } } Job Announcement: } } } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } } } UNL Center for Materials Research and Analysis } } } } } } Analyze/characterize materials using electron microscopy, materials } } } preparation and computer instrumentation. Supervise/train students, } } } faculty and visiting researchers utilizing these methods. Bachelor's with } } } major in physical science, engineering or related field plus three years } } } experience in the operation, repair or design of electron microscopes or } } } other scientific instrumentation. Master's preferred. Must have excellent } } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } } diffraction or materials sample preparation experience preferred. Excellent } } } benefits. Submit cover letter, resume and names, addresses and telephone } } } numbers of three professional references to Professor Brian Robertson, } } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } } will remain open until a suitable candidate is found. UNL is committed to } } } AA/EEO and ADA/504. If you require accommodation, please call (402) } 472-7886. } } } } } } } } } *********************************************************** } } } Assoc. Prof. Brian W. Robertson } } } Department of Mechanical Engineering } } } and Center for Materials Research and Analysis } } } University of Nebraska-Lincoln, N124 WSEC, } } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } } } Cheers, } } Gary Gaugler, Ph.D. } } } } } } } } } } } RFC822 header } ----------------------------------- } } Received: from Sparc5.Microscopy.Com [206.69.208.10] by mailhouse.hei.org } (SMTPD32-4.07) id A18E691011E; Fri, 14 May 1999 13:03:26 PST } Received: (from daemon-at-localhost) by Sparc5.Microscopy.Com (8.6.11/8.6.11) } id IAA02338 for dist-Microscopy; Fri, 14 May 1999 08:40:47 -0500 } Received: from no_more_spam.com (Sparc5 [206.69.208.10]) by } Sparc5.Microscopy.Com (8.6.11/8.6.11) with SMTP id IAA02333 for } "MicroscopyFilteredEmail-at-msa.microscopy.com"; Fri, 14 May 1999 08:40:16 -0500 } Received: from poteidaia.utdallas.edu (poteidaia.utdallas.edu } [129.110.10.1]) by Sparc5.Microscopy.Com (8.6.11/8.6.11) with ESMTP id IAA02322 for } {Microscopy-at-sparc5.microscopy.com} ; Fri, 14 May 1999 08:40:03 -0500 } Received: from apache.utdallas.edu (kna101-at-apache.utdallas.edu [129.110.16.9]) } by poteidaia.utdallas.edu (8.9.1/8.9.1/null-3.5) with ESMTP id IAA14724; } Fri, 14 May 1999 08:42:30 -0500 (CDT) } Received: from localhost (kna101-at-localhost) } by apache.utdallas.edu (8.9.1/8.9.1) with SMTP id IAA01748; } Fri, 14 May 1999 08:42:29 -0500 (CDT) } X-Authentication-Warning: apache.utdallas.edu: kna101 owned process doing -bs } Date: Fri, 14 May 1999 08:42:29 -0500 (CDT) } From: Karen S Pawlowski {kna101-at-utdallas.edu} } To: "Dr. Gary Gaugler" {gary-at-gaugler.com} } cc: MSA listserver {Microscopy-at-Sparc5.Microscopy.Com} } Subject: Re: Electron Microscopy Position Available Immediately } In-Reply-To: {4.1.19990513195604.009dc490-at-pop.calweb.com} } Message-ID: {Pine.GSO.3.96.990514083332.1144A-100000-at-apache.utdallas.edu} } MIME-Version: 1.0 } Content-Type: TEXT/PLAIN; charset=US-ASCII } Errors-to: Microscopy-request-at-sparc5.microscopy.com } X-UIDL: 907884271 } Status: U }
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Actually this salary range is not as low as you would think.. I have a M.S. degree and 5 years of EM experience and it took a while for me to find a job in EM throughout the country.. I was applying for jobs that were offering $25K to start as a salary and some were more depending on what part of the country you were looking in... I ended up in Seattle at a job for $32K with benefits to start... I am now currently in Los Angeles at UCLA Medical Center and the salary here is quite a bit more than in Seattle.. But that is outweighed due to the high cost of just living here...So the salary for this job is not as bad as one might think...
==========================
} You are not the only one shocked about this. Looking at the figure as low } as Mid-$30K, I would say there is something seriously not quite right. } These days a simple graduate in computer programming with 2-3 years } experience easily gets a starting salary of $50k+ and as they go ahead with } few more years of experience, they quickly move into the scale of $70k-80k } and crosses $100k within a span of well less than 10 years of career. } } - Is there something going wrong with microscopists and SEM/TEM engineers? } - Is Mid-$30k fair to expect for electron microscopists (3 years } experienced) who maintains the equipment as well? } - What if they have 12 -15 years of experience? } - Is $60k max (typically at Universities) is fair that these gorgeous } people are expected to get max. in their lifetime? } - Is that what the people who are the backbone for running and maintaining } research facilities are supposed to rewarded with? } } I mean just compare it with professionals with same qualifications and } experience in other fields. In my personal opinion, there is something not } quite right and needs to be re-assessed. } } Zia ur Rahman } University of Central Florida, } Orlando, Florida } } } At 07:59 PM 5/13/1999 -0700, you wrote: } } } Mid-$30K? Is this for real? Is this the going rate for SEM } } specialists? That is about 1/4 of what I would consider. } } There must be some subtle differentiation of what is expected } } from such positions vs. the qualifications and experience of } } others in the field. Am I the only one shocked about this? } } } } } } } } } JOB ANNOUNCEMENT BELOW -- PLEASE POST } } } } } } Dear colleagues, } } } We want to hire a suitable person for the Central Facility for Electron } } } Microscopy at the Center for Materials Research and Analysis at the } } } University of Nebraska-Lincoln (UNL). The salary will at least likely be in } } } the range of the mid $30k's and is dependent on experience. The medical, } } } dental and retirement benefits package is substantial and includes } } } subsidized tuition for employees to take UNL courses. (BTW, Lincoln is } } } growing, has low unemployment, is great for families, has a good range of } } } live music, dance and theater, and has very good public and other schools.) } } } The Facility provides user-access for ~60 faculty plus their students and } } } other researchers. The Facility includes a JEOL 2010 TEM, a JEOL 840A SEM } } } and a VG HB501 field-emission STEM, along with a full range of accessories, } } } specimen preparation equipment, computers and darkroom. Some development } } } of new research instrumentation is in progress for mapping of magnetic } } } materials and more development is planned for the Facility. You can find } } } out more about the Facility on our web site (that does need a little work) } } } at URL: http://www.unl.edu/CMRAcfem/ } } } Thanks for passing on the word, } } } Brian Robertson } } } } } } ********************************************************** } } } Job Announcement: } } } } } } MATERIALS MICROSCOPY RESEARCH SPECIALIST } } } } } } UNL Center for Materials Research and Analysis } } } } } } Analyze/characterize materials using electron microscopy, materials } } } preparation and computer instrumentation. Supervise/train students, } } } faculty and visiting researchers utilizing these methods. Bachelor's with } } } major in physical science, engineering or related field plus three years } } } experience in the operation, repair or design of electron microscopes or } } } other scientific instrumentation. Master's preferred. Must have excellent } } } computer and interpersonal/communication skills. TEM, SEM, x-ray } } } diffraction or materials sample preparation experience preferred. Excellent } } } benefits. Submit cover letter, resume and names, addresses and telephone } } } numbers of three professional references to Professor Brian Robertson, } } } CMRA, N104 Walter Scott Engineering Center, UNL, Lincoln, NE 68588-0656 or } } } e-mail brobertson-at-unl.edu. Review of resumes will begin June 1. Position } } } will remain open until a suitable candidate is found. UNL is committed to } } } AA/EEO and ADA/504. If you require accommodation, please call (402) } 472-7886. } } } } } } } } } *********************************************************** } } } Assoc. Prof. Brian W. Robertson } } } Department of Mechanical Engineering } } } and Center for Materials Research and Analysis } } } University of Nebraska-Lincoln, N124 WSEC, } } } 17th & Vine Sts., Lincoln, NE 68588-0656, USA } } } ** Office 402 472 8308; FAX 472 1465; Labs 472 8762 and 472 5498 ** } } } } Cheers, } } Gary Gaugler, Ph.D. } } } } } } } } }
no creo que seamos muchos los que hablamos espanhol en esta lista, te recomendaria que escribieras en Ingles y asi ampliar tus posibilidades de obtener ayuda.
Elena,
I dont think that many of us speak spanish, I will recommend you to write in english, increasing, in this way, your possibilities of obtainig help from this list.
saludos
Gary=20 EPhD Trondheim, Norway.
On Tue, 27 Apr 1999, Elena Brandaleze wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America= =20 } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } =20 } =20 } Sres MSA: } Dado que mi tema de investigaci=F3n es el an=E1lisis de propiedades } mec=E1nicas de pol=EDmeros semicritalinos, necesitar=EDa contactarme con } alguna persona que pueda informarme sobre estudios de microestructura de } los mismos.Desde ya muchas gracias.=20 } =09=09=09=09Ing elena Brandaleze. } =20
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Hi, The subject line says all: I am using LM to look at sections embedded with Spurr's, and I would like to know its refractive index. Haven't been able to track it down on the web or through my vendor.
Thanks, Jill
------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
} Date: Sun, 16 May 1999 14:42:53 +0000 } To: Paul Webster {pwebster-at-mailhouse.hei.org} } From: Lou Solebello {microls1297-at-mindspring.com} } Subject: Re: Electron Microscopy Position Available Immediately } In-Reply-To: {199905142059.PAA08933-at-Sparc5.Microscopy.Com} } } What is expected of an electron microscopist, or ANY microscopist for that matter (and I use that term loosely....there are a lot people who carry the title but are nothing more than a user), is DIRECTLY relate to what they get paid. } } The amount of responsibility that we microscopist take on relative to other professionals in other fields is a great deal under compensated. We are expected to know everything there is about our individual discipline as well as all other aspects of microscopy. We are expected to deliver THE ANSWER nearly immediately at a low cost. We are expected to know how to assemble and dissasemble our instruments and fix them at the same cost and rate. We are looked upon sourly when we say "cant be done" as if we are inferior or lying. We are expected to handle samples that potentially are extremely hazardous to our health.....the danger of many unknowns on the consulting level are unknown until we discover what we are analyzing. We are expected to write SOP's, develop QA/QC manuals, submitt contract bids, obtain grant funding, supervise people, be walking encyclopedia, be able to read client minds, maintain proficiency in round robins and regulatory complance audits, be an MBA and business wiz, and work 60 hours a week on salary if required at a moments notice. } } That is the real point that I have been trying to get across to others on this list server. We are GROSSLY underpaid considering our responsibilities, level of education and experience. The only thing that keeps most of us in these positions is the fact that we love our work. Nobody said life was fair.... } } At 02:06 PM 5/14/99 -0700, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
John writes ... } } } I know there have been some discussions on this listserver in the past } about computer sign up systems for electron microscopes. Are there any new } programs out there? We are working on a web-based instrument sign up } program to replace our existing sign up/microscope accounting system. The } old system is not Y2K friendly. Besides the Universty of Minnesota, is } there any other group which has a working web-based system?
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Well folks.... I screwed up and during a reprogramming of the Junk Mail Filter managed to muck up the works. I did not notice that the "Listserver" Mail was off-line and inoperative until this evening ~ 10pm, since the rest of my mail was operating fine.
I hope you all enjoyed your present of "peaceful" day.
If you tried posting a message and it was rejected or you just did not see it, then just try again. I think all is back to the status quo.
Cheers... Nestor Your Friendly Neighborhood SysOp.
PS
Hmmm.. I wonder how many people noticed ? Naw.. I won't ask..
We have some spare equipment to dispose of to a good home, all in good working order.
Prices are negotiable, but you will have to collect.
1. De Vere 504 enlarger
2. Durst M35 colour enlarger
3. LKB knifemaker 7800 series (sensible offers on this one please)
4. Reichert OM U3 ultramicrotome
Hurry while stocks last!
Pete
Peter Bond Plymouth Electron Microscopy Unit University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel/Fax: 01752 233092 email: pbond-at-plymouth.ac.uk
A colleague of mine needs to replace an old keyboard for a Tracor Nothern TN 5500 EDS system. Does anyone have one to sell trade or give away. Please contact Charlie Cooney -at- CBC-at-post.queensu.ca Thank you Paul Nolan
I am looking for individuals interested in cryoplaning techniques (using a cryoultramicrotome to produce blocks with a very flat surface and examining this surface in a cryo-SEM). Please contact me back-channel if interested.
Best regards, Steven E. Slap, Vice-President ******************************************* Energy Beam Sciences, Inc. The Laboratory Microwave Company Adding Brilliance to Your Vision http://www.ebsciences.com {http://www.ebsciences.com} *******************************************
Wyeth-Ayerst Research, a major division of Fortune 100 American Home Products Corporation, has an opportunity at our pharmaceutical research facility in Chazy, New York for a Scientist in our Investigatory Pathology group.
JOB TITLE: Scientist II, JOB# C5780 DEPARTMENT: Investigatory Pathology LOCATION: Chazy, NY HR REPRESENTATIVE: B. Hebert HOURS: 8:00 a.m. - 4:30 p.m. POSITION REPORTS TO: Dr. Laura Patrone, Senior Research Scientist I
EDUCATION REQUIREMENTS: BS/BA degree in Biology, Biochemistry, or science related field or MS degree in science related field.
EXPERIENCE REQUIREMENTS: BS/BA candidates must have a minimum of 4 years experience developing immunohistological assays in an industrial, hospital or highly specialized academic setting; MS candidates must have a minimum of 2 years relevant pharmaceutical laboratory experience, or 2 years experience in a highly specialized relevant academic environment.
OTHER SKILLS REQUIRED: Additional histomorphological skills required; must have excellent oral and written communication skills as well as excellent interpersonal and organizational skills; some travel may be required.
PRINCIPAL DUTIES OF POSITION: Scientist II will perform immunohistochemistry and associated techniques, develop new panels of tests to characterize tissues and differentiate cell types; will provide technical support for method development in morphologic analytical techniques; perform data interpretation and assist in report writing; review literature for new technologies to expand and enhance research capabilities; will provide technical support for regulatory pathology area; will also perform departmental and facility support functions as needed (e.g., inventory, scheduling, purchasing).
Interested candidates must contact the Human Resources Representative: Barbara Hebert Office of Human Resources Wyeth-Ayerst Research 641 Ridge Road Chazy, NY 12921 Phone (518) 846-6237
Available - Balzers 360M Freeze Fracture System, 1970 vintage. Complete with E-Beam Pt-C gun, evaporative C electrodes, QSD-201D quartz crystal thickness monitor, commutator unit, and timer for C electrodes. Also included is extra EVM-052A E-Beam power supply, extra GA-1 control unit, extra ODP, extra main valve, extra baseplate, microtome, and feedthroughs, and many extra sample holders, feedthroughs, tools, etc. Must move immediately for best offer. Robert J. Munn rjmunn-at-ucdavis.edu
I am looking for a method to visualize the somata of live epithelial cells. I plan to inject these cells intracellularly, using a micropipette. What I need is a stain with the following characteristics: 1) it can be injected intracellularly via a micropipette, preferably using intophoresis. 2) it is NOT fluorescent 3) it is visible without the need to further process the tissue. This last point is important because I need to watch the injection under the LM and be able to tell when the cell is filled. --E. Peterson Ellengene H. Peterson Neurobiology Program & Department of Biological Sciences Ohio University Athens, OH 45701 740 + 593-2111 (tel) 740 + 593-0355 (fax) peterson-at-ohiou.edu
Select the online scheduler; username=guest, password=guest. You will be able to make reservations on the instrument "Dummy".
Unfortunately not all web browsers are created equal. You can use Navigator 4 (or higher) or IE 4 (or higher) on a PC, but only IE 4 (or higher) on a Mac. (It is a Java compatibility problem). You will also have to accept a cookie so that the server can recognize the platform you are running on.
cheers, Henk
At 06:15 PM 5/16/99 -0700, you 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://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
Hi everyone, First of all I would like to thank Dr. Simkin from Michigan University forthe information about Electron Channelling. I am going to get in touch with him. This time my question is about jet polishing solution of copper and copper and zinc alloys. Actually i found a solution that is sixty percent phosphoric acid and deionized water but the problem is that it is hard to get rid of that solution from the surface of specimen after you are done with jet polishing. If anybody has some idea about how to clean the surface after jet polishing i really appreciate it. Thanks
I'm looking for any input on the quality of Russian made LOMO (Leningrad Optical and Mechanical Organization) microscopes. Specifically the Biolam, Polam models and the stereomicroscope marketed here in the US.
I would be interested to hear from anyone who has experience using these scopes and commenting on quality of optics, mechanical, illumination,etc. Although you get alot for your money, I'm concerned about service, spare parts, and future support.
Much appreciated.
{italic} {color} {param} 8080,0000,0000 {/param} Raymond Nip, RRT, RCP
I would like to begin by telling you I work for and represent a manufacturer, Allied High Tech Products, Inc. We manufacture the MultiPrep=99 system.
The MultiPrep is a semi-automatic tool used for many applications including TEM sample preparation using the wedge technique. The MultiPrep's= =20 capabilities include parallel polishing, precise angle polishing (0.02 degree increments), site-specific polishing or any combination thereof. It provides reproducible results by eliminating inconsistencies between users. When using the MultiPrep only the specimen is in contact with the abrasive, eliminating the problem of excessive wear of the hand tool's feet, and unwanted faceting of your specimen. In fact, by using the MultiPrep, you can completely eliminate the need for hand-held polishing tools.
The MultiPrep allows you to set an angle and polish through the angle without it changing, and monitors the amount of material that has been removed from the sample (real time during the polishing operation) in 1-micron increments. =20
The MultiPrep system sells for $11,775.00 plus the accessories that are necessary for your individual applications. =20
The other applications of the tool are preparation of SEM cross sections, TEM sample Prep, Pre-FIB thinning, parallel polishing and de-layering of semiconductor devices or thin section preparation and backside sample preparation for semiconductor devices for emission spectroscopes.
If you have any questions please feel free to contact me, send mail to mailto:info-at-alliedhightech.com, or visit our web site at http://www.alliedhightech.com.=20
We also have an new 12 page brochure that can be emailed to you in a PDF format. It can be sent via snail mail too.
Thank you,
Ed Hirsch
} Hello } } We are to begin the process of using tripod polishing for preparing } TEM samples, and request vendors to contact me with regards to } types of polishers available, style, cost, ease of use. } } I also welcome users of this method to comment online (or email directly) } as to the ease of use and technique of sample preparation, since our lab } has not gone this route before. } } thanks in advance } } Fred } } } } } } ******************************************************** } Fred Pearson } Brockhouse Institute for Materials Research } McMaster University } 1280 Main St. West } Hamilton, Ontario=20 } Canada L8S 4M1 } } } email: eoptics-at-mcmaster.ca } phone: (905) 525-9140 ext. 24609 } fax: (905) 521-2773 } ******************************************************** } } } ************************************************* Edward A. Hirsch Product Application Specialist Allied High Tech Products 2376 East Pacifica Place Rancho Dominguez, CA 90220 ph: (919) 846-9628 vm:(800)675-1118 x245 fx: (310)762-6808 http://www.alliedhightech.com *************************************************
I have a JEOL JWS 7515 in England that need to be de-installed. This is a Critical Demension SEM. Does anyone have a lead on independent service companies that can do this type of work?
Regards,
Earl Weltmer Scanservice Corporation Third Party SEM Service USA
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Hi, Zia: The OL coil is OK for three reason: 1. you have told me that you found a short so that the OL coil was unconnected to the connector. It was meaning you found out the reason why the OL coil is open. 2. If the OL coil was broken or short inside, the OL resistance would be changed. But you know 5.2 ohms is OK. 3. The insulation of OL coil is relative with high voltage and the temperature of the OL coil only. If you keep it below 80 C degree, you can test it by DC power supply and flow 10 amps currents in 10 minutes . In fact, you have get right way and go on please . I do not know why you can not receive my e-mail and you can call me 65-8743253 ( office phone) at 9:00 pm Singapore time. Good luck! Regards Tiejun
to add a comment on the answers you've already got concerning your request on measurements of optical parameters for HRTEM simulation... Mike O'Keefe's answer is acceptable as a first try. If you want to measure some values more accurately (especially the Cs), just refer to the literature quoted by Jonathan Barnard. However, there is a little problem with the Cs measurement from HRTEM diffractograms obtained from an amorphous thin film ; the usual method assumes that the specimen behaves like a pure weak-phase object, which is generally not fulfilled : there is a phase shift due to the propagation of electrons throughout the object, which leads to a breakdown of the 'potential projection' hypothesis, which may lead to false values of defoci and Cs... To get more details on this problem raised by J.M. Gibson (Ultramicrosc., 1994), check the work below (where the complete reference from J.M.G. is also recalled) :
"Quantitative analysis of HRTEM images from amorphous materials. I : about the estimation of Cs and df from HRTEM diffractograms", H.S. BAIK, T. EPICIER, E. VAN CAPPELLEN, Eur. Phys. J. AP 4, 11-26, (1998)
As a consequence of the approach developed within this paper, you can also estimate rather precisely the other parameters (delta and alpha) if you know precisely the structure (and the thickness) of the amorphous film you're using for your measurement ; the treatment is however somewhat tedious, but it works (we have applied it successfully to such measurements for a JEOL2010F and a CM200FEG).
Good luck,
--------------------------------------------------------------------------- Dr. Thierry EPICIER, GEMPPM, umr CNRS 5510, INSA de LYON, Bat 502, 20, Av. Einstein, F69621 VILLEURBANNE CEDEX FRANCE
I need to characterise the microstructure and composition of soft electroceramic film(PZT) by using FEGTEM. When I put the specimen in to the beam path, I observed that there was a very strong interaction between the electron beam and specimen which can clearly be seen to be badly charged. I have been advised to coat carbon or gold on the surface of the specimen. I would like to know if there any other methods which could be used to avoid this charging problem? Do you have any further ideas or methods for this sort of material to prepare a specimen which is suitable for high resolution image and electron energy loss spectroscopy? Currently, I use a method of ion thinning by Gatan DuoMill and PIPS.
Dear members: I have placed an order for a dimpler and precision coring tool made by VCR company several months ago and am now waiting for their shipping. But I was told that VCR was bancrupt or will be sold by other company. I worry about the quality of the product and A/S. Could any of you send me the right story what is going on there? Any information will be appreciated. Best wishes, Jondo YunDepartment of Inorganic Materials EngineeringCenter for Instrumental AnalysisKyungnam University449 Weolyeong-dong, Masan, 631-701, Korea82-551-249-2697 (tel)82-551-248-5033 (fax)jdyun-at-hanma.kyungnam.ac.kr
A quick question: we have some blood samples which were received in "Trump's Fixative", described as phosphate-buffered gluteraldehyde and paraformaldehyde, pH 7.2. So far we've been unable to locate any reference to Trump's in any of our literature.
I expect it's perfectly fine to proceed with regular phosphate buffer washes, then postfixation, but thought I would check with collective microscopy mind out there first.
Earl Weltmer wrote: =============================================== I have a JEOL JWS 7515 in England that need to be de-installed. This is a Critical Dimension SEM. Does anyone have a lead on independent service companies that can do this type of work? ============================================== You can find listings for third party SEM service firms in the UK on our website URL http://www.2spi.com/hot-service5.html
This is part of our website directory of third party service providers on equipment in the microscopy and microanalysis world.
Either one of them could probably do what you want to have done.
Chuck
Disclaimer: We have no financial interest in either firm, both have good reputations, so far as we know.
=================================================== 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: www.2spi.com ############################ ==================================================
I have noticed that some non-conductors can be put into the microscope without the objective aperture and they can set up a steady state where they do not charge. Once you put the objective aperture in they continually charge. I recently noticed on a GaN on sapphire substrate imaged in a 120 kV machine where this behavior is reversed, i.e. charges without the aperture in and does not when it is in.
I have had some success with the small angle cleavage technique with glass substrates without coating. Most times the sample will not charge without coating, but some times it will. I try to minimize the amount of material potentially exposed to the beam by putting the silver epoxy as close to the tip area as possible.
However, I am using a 120kV machine for most of the time and I need to carbon coat to keep the glass from softening. I have less of a problem with this in a 200kV machine. I believe that there is more heating in the sample at 120kV than 200kV because more current is being deposited into the sample at the lower voltage. At higher voltages more electrons pass through the sample.
Incidentally, when I do coat with carbon, I can get by with about 200 A on each side. You can get away with about 400 A on one side but there are other reasons why I need to coat on both sides. This thickness is about the minimum for me to prevent softening of the glass. I think that you can get away with only about 200 A or less on just one side if your sample doesn't have this problem. There was a question some time ago that I asked when I first started working with glass about whether it matters if you coat just one side or two and if you coat one side does it matter if it is towards the beam. I think that you get a little better performance with the coating towards the beam, especially if the beam is exposed to any thick parts of the sample at lower magnifications.
These observations are all based on a limited experience of a little over a year on glass samples.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: P.Wang To: Microscopy-at-Sparc5.Microscopy.Com -----------------------------------------------------------------------.
Hi all
I need to characterise the microstructure and composition of soft electroceramic film(PZT) by using FEGTEM. When I put the specimen in to the beam path, I observed that there was a very strong interaction between the electron beam and specimen which can clearly be seen to be badly charged. I have been advised to coat carbon or gold on the surface of the specimen. I would like to know if there any other methods which could be used to avoid this charging problem? Do you have any further ideas or methods for this sort of material to prepare a specimen which is suitable for high resolution image and electron energy loss spectroscopy? Currently, I use a method of ion thinning by Gatan DuoMill and PIPS.
That is the way we do it all the time. You can find info on Trumps at this site by searching for such.
http://www.biotech.ufl.edu/~emcl/tips.html
At 08:23 AM 5/19/1999 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Don't you get Calcium Phosphate precipitates forming if you fix in a phosphate buffered aldehyde solution???
} A quick question: we have some blood samples which were received in "Trump's } Fixative", described as phosphate-buffered gluteraldehyde and } paraformaldehyde, pH 7.2. So far we've been unable to locate any reference } to Trump's in any of our literature. } } I expect it's perfectly fine to proceed with regular phosphate buffer } washes, then postfixation, but thought I would check with collective } microscopy mind out there first. Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility
3 Tucker Hall Division of Biological Sciences University of Missouri Columbia, MO 65211-7400 (573)-882-4712 (voice) (573)-882-0123 (fax)
} From September 1999 there will be a 2 year post-doc position available in the electron microscopy group (Department of Physical Chemistry) at the University of Mainz, Germany. At present the group has 2 electron microscopes and a well-equipped specimen preparation laboratory. In September 1999 a new 300 kV Philips Tecnai 30 Electron Microscope with FEG and GIF will be installed. Applications from physicists and chemists with Ph.D experience in electron microscopy and a sound background in electron energy loss spectroscopy are welcome.
Please send applications to
Dr. I.G. Voigt-Martin Inst. f. Physikalische Chemie Jacob Welder Weg 11 55099 Mainz
Responding to the message of {4.1.19990518173013.030d9100-at-pop.er6.eng.ohio-state.edu} from "Hendrik O. Colijn" {colijn.1-at-osu.edu} :
} Feel free to check out our site. } } http://www.ceof.ohio-state.edu }
I tried, but it doesn't seem to like the single most popular browser/platform combination at our University - Netscape Navigator running on Mac OS.
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist Office:(612) 626-7594 CIE Characterization Facility, University of Minnesota Desk: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
Hi, I am looking for a low magnification oil immersion objective. I look at two scales in my work -- I use 100x oil for locating bacteria and 25x for the sediment grains they are attached to, and I need to be able to go back and forth. I do not need a high NA. This is for a Zeiss Universal, non-infinity corrected. From what I gather, older Leitz and perhaps olympus objectives are interchangeable. Thanks if you can help, Jill
------------------------------------------------------------------------------ Jill L. Schmidt Phone: (206) 685-1926 University of Washington FAX: (206) 543-0275 Oceanography Box 357940 e-mail: jills-at-ocean.washington.edu Seattle, WA 98195-7940 http://www.ocean.washington.edu/people/grads/jills/Schmidt.html ------------------------------------------------------------------------------
OK, I know asking this question of the group may seem strange, but this is the most densely populated area of microscopy wisdom in the universe and I need your global perspective.
I am the only person in a small EM lab that serves a primarily undergraduate campus. I am part of the technical staff, I am not a faculty member. We have doctorate programs and a few researchers using TEM, but no materials science types and no professional schools. I guess our TEM might be used 300 to 500 hrs/yr sometimes less, never more.
We have a JEOL 100B circa 1975. For most of our applications, it works fine. I have a whole other microscope to use for parts and a clever service provider who may be able to keep this thing going for a long time. In a darkened room, it is hard to tell it from a more modern instrument based on the images seen on the screen. When the lights go on, it looks 24 years old.
Recently a new faculty member asked when are we getting a new TEM, one with digital imaging. I choked, said I would check into it. So here are a few questions I need help with and I hope for some kindness and understanding in your replies. I do need your replies because as the only real EM person here I could use your help.
I have never gone out looking for $$ for a new microscope. Where does one look? Could we justify $$ based on our level of usage? What do you think? What are the criteria used by funding agencies when evaluating requests. Could you share you experiences?
Suppose we can't get the $$ for a whole new TEM. Would I be crazy to try to get $$ to equip our current TEM (remember 24 years old) with an add on digital camera? The ports are there and it could be done, but I'm not sure it is a good idea.
I hope you can give me some leads and encouragement. Most of the burden of doing this job will fall on me. Although often supportive in casual discussions, I don't expect much help from anyone else here.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I have looked in our archives and have found 2 solutions that Bernie Kest= el from Argonne National Lab has developed for copper which may be of use. =
This work was done with our Model 550 single vertical jet electropolisher=
so references to jet height would be unique to our system. The other parameters may need to be adjusted slightly depending on the type of jet polisher you are using. =
Recipe #2 150 ml HNO3 350 ml ethanol 40 ml butyl cellosolve
Temp: -20 degrees C Jet height: 3.9mm Pump setting: 2.5 Volts: 40 Current: 75mA NOTES: excellent polish. Lower temperature result in an uneven foil surface.
I hope this information helps. If I can be of any additional assistance,=
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by Ozgul Keles } ------------------------------------------------------------------------=
The Microscopy ListServer -- Sponsor: The Microscopy Society of America =
Hi everyone, First of all I would like to thank Dr. Simkin from Michigan University forthe information about Electron Channelling. I am going to get in touch=
with him. This time my question is about jet polishing solution of copper and copper and zinc alloys. Actually i found a solution that is sixty percent=
phosphoric acid and deionized water but the problem is that it is hard t= o get rid of that solution from the surface of specimen after you are done with=
jet polishing. If anybody has some idea about how to clean the surface after jet polishing i really appreciate it. =
Does your current TEM meet the department's needs?
You said: "no materials science types and no professional schools." are you into biological TEM, engineering or physics?
A bit more information is needed to help you on the technicality.
As for age, I am not sure it is such a problem. We have a Philips EM400 (120 keV) bought back in 1984 (I think- I was 11 at the time!), which despite its age is fantastic to work with. The success of this machine is that we have always had a service contract with Philips on it. If your machine is fine at the moment why don't you get a good service contract, and maybe spend some money on retro fitted equipment, for example CCD camera for digital imaging.
If you need to extend the TEM capability you may want to invest in a second hand machine ($10,000's) that will be a bit more adaptable. If you want to do this, then I am sure someone will suggest a web page, so that you can have a look at the current price ranges. A safer bet is to contact a reputable TEM manufacturer (Philips, Jeol, Hitachi, .etc) and ask for a contact and ask if they will support it.
I am afraid you are going to have to do a bit of research about this, so it may take a couple of weeks to get an idea as to what your options are. If you do this you should find yourself with a TEM that is well supported (if things do go wrong), should be a delight to use and be at a good price. However if you don't you could end up with something that is a waste of cash, frustrating to use and out of date.
Good luck, Jon
P.S. I know at present Philips are taking a lot of orders for their new Tecnai's (fully integrated computer/TEM), so you may find that there are a lot of second hand TEMs being advertised over the next few years.
-- ***************************************** Dr Jonathan Barnard
Microstructural Physics, H.H.Wills Physics Laboratory, University of Bristol, Tyndall Avenue, Bristol BS8 1TL.
In a message dated 5/19/99 2:25:49 PM EST, jmkrupp-at-cats.ucsc.edu writes:
{ { I guess our TEM might be used 300 to 500 hrs/yr sometimes less, never more.
We have a JEOL 100B circa 1975. For most of our applications, it works fine. I have a whole other microscope to use for parts and a clever service provider who may be able to keep this thing going for a long time. In a darkened room, it is hard to tell it from a more modern instrument based on the images seen on the screen. When the lights go on, it looks 24 years old.
Recently a new faculty member asked when are we getting a new TEM, one with digital imaging. I choked, said I would check into it. So here are a few questions I need help with and I hope for some kindness and understanding in your replies. I do need your replies because as the only real EM person here I could use your help.
I have never gone out looking for $$ for a new microscope. Where does one look? Could we justify $$ based on our level of usage? What do you think? What are the criteria used by funding agencies when evaluating requests. Could you share you experiences?} }
The decision to buy another instrument would depend entirely on need for additiona; features. Sounds as though the current instrument works well for your current needs. However, you might consider upgrading to a more recent instrument with additional features - but look for a used EM. There always seems to be used EMs available at excellent prices.
{ {Suppose we can't get the $$ for a whole new TEM. Would I be crazy to try to get $$ to equip our current TEM (remember 24 years old) with an add on digital camera? The ports are there and it could be done, but I'm not sure it is a good idea.} }
If you have the ports then why not. So long as you have a good resolution image coming down the column then you can do what you want with it in terms of image storage.
The Department of Microscopy and Microanalysis at Abbott Laboratories i= s recruiting a senior level microscopist for its Materials Science group.=
Requirements include a Ph.D. in Materials Science, Chemistry, or relate= d field, and a working knowledge of most of the following instrumentation= : TEM, SEM, EDXS, XPS/SIMS, AFM, and light microscopy methods (polarized light= , fluorescence, and confocal microscopy). Familiarity with spectroscopic=
techniques such as FTIR and a strong background in physical and analyti= cal chemistry are desirable. Experience with biological systems would be = highly advantageous.
We are looking for a team player with excellent interpersonal skills an= d the ability to readily adjust to rapidly changing priorities. The successf= ul candidate will seek out and learn new technologies as needed to solve problems related to pharmaceutical and healthcare products. The abili= ty to communicate clearly, both verbally and in writing, is essential.
Essential job functions: Independently design and carry out experiments to evaluate materials specimens by microscopic and microanalytical techniques. Develop methods using multidisciplinary approaches. Interpret data and effectively communicate results. Review data and reports for scientific integrity and clarity. Mentor career development of junior scientists. Present data at scientific meetings and publish in peer-reviewed journa= ls. Provide technical and scientific leadership.
The Department of Microscopy and Microanalysis provides corporate-wide support in materials and biological microscopy to all Abbott Laboratori= es divisions. The facility houses two TEMs (a Philips CM12 STEM and a LEO= 910), two SEMs (a Philips XL30 and AMRAY 1830i), three EDXS systems, a BioRad= MRC 1024 UV confocal scanning laser microscope, a Digital Nanoscope III AFM= with Bioscope, a Physical Electronics 5600 XPS/SIMS, and the usual assortmen= t of light microscopes and sample preparatory equipment.
For further information, please contact:
Jane A. Fagerland, Ph.D. Abbott Laboratories D45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202
Hello dear all! I am looking for any information on the microscope, called Grumhauser and on the person who made it. The spelling I have might be incorrect. Best regards, Dima My email address is la_dima-at-hotmail.com
_______________________________________________________________ Get Free Email and Do More On The Web. Visit http://www.msn.com
You can test the reservation system by logging in as user=guest, password=guest. You can make reservations on the "dummy" machine. During the couse of writing the software, we discovered that not all web browsers are created equal. There seem to be some major differences in the level of Java support. Anyway, on a Mac you need IE 4.0 or higher, on a PC you need IE 4.0 or Navigator 4.0 or higher Mac Navigator doesn't work yet.)
Our system is running on a WinNT PC using Microsoft Access as a database backend. The front end was done with MS InterDev 6.0 and Borland JBuilder 2.0.
At 10:33 AM 5/15/99 -0500, you wrote: } } } We are also considering developing a web-based reservation system, and I } think many others would also be interested what other labs are doing in } this area. I would be interested in learning of any software which is } freely available or low cost and open source. } } We are in the very early stages of modifying an open source (Perl) } web-calender and scheduling system to suite our needs. The reference url } is: } http://curiosityshoppe.tierranet.com/framecal/index.shtml } } I found this and a few others by searching the web with I believe was } combinations of ("resource" and "scheduling" and "calenders") } } If we develop a system based on FrameCal or something else we will make } it freely available as licensing permits. } } Of course, if someone else has a suitable system available freely or at } low cost it would save us development time and costs. } } Jim Mabon } } } } } } I know there have been some discussions on this listserver in the past } } } about computer sign up systems for electron microscopes. Are there any } new } } programs out there? We are working on a web-based instrument sign up } } program to replace our existing sign up/microscope accounting system. } The } } old system is not Y2K friendly. Besides the Universty of Minnesota, } is } } there any other group which has a working web-based system? } } } } John C. Wheatley } } Lab Manager } } Arizona State University } } Center for Solid State Science } } PSA-213 } } BOX 871704 } } Tempe, AZ 85287-1704 } } } } Phone: (602) 965-3831 } } FAX: (602) 965-9004 } } John.Wheatley-at-ASU.Edu } } } Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility 116 W. 19th Ave. (614) 292-0674 "Nothing is as inevitable as a mistake whose time has come."
I have some crystals of a drug in aqueous suspension. They clump together. I would like to disperse the crystals. Sonication is about the only way I can think of to do it. Any ideas? If sonication, for how long should I do it (about 0.3 ml volume) and how long would it be likely to last before it all clumps again? I should add that I can't add anything to the solution and I don't know the chemistry of the drug!
Thanks, Diana
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
At 12:10 PM 5/19/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Consider other than a new scope. Digitize your existing scope for a fraction of the cost of a new scope. A good source is http://www.elmdas.com
They make a nice computer control and image analysis package at a decent price. POC is John Best.
One- or two-year POST-DOC position available at the Electron Microscopy for Materials Science (EMAT) group of the University of Antwerp, RUCA, Belgium. Do visit our website for more info on the working of the group (http://wcc.ruca.ua.ac.be/EMAT).
The position is part of a European TMR network (9/98 - 9/02) on Phase Transitions in Crystalline Solids and is open to inhabitants of member states of the European Community or associated countries. The network is a collaboration between experimental and theoretical groups all working on the understanding of microstructures resulting from phase transitions in solids (see also http://www.dmsa.unipd.it/tmr/). Subprojects on martensitic transformations in alloys, twinning in oxides, thin films structures etc. have already been chosen. The candidate should have a large experience in different TEM techniques for the characterisation of atomic and microstructures. The position is open as of October 1, 1999. Salary starts at 1.500 Euro (scholarship, no taxes deducted), depending on experience.
If you are interested or like more information, please contact Dr. D. Schryvers (tel.: 32-3-2180247, fax: 32-3-2180257, e-mail: schryver-at-ruca.ua.ac.be)
!!!!! NEW SERVER !!!!
DO CHECK MY E-MAIL ADDRESS: schryver-at-ruca.ua.ac.be (replies to old mails, e.g. from before March 15, will not work anymore!)
*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=* *=* *=* *=* Dr. D. Schryvers *=* *=* Electron Microscopy for Materials Research (EMAT) *=* *=* University of Antwerp, RUCA *=* *=* Groenenborgerlaan 171 *=* *=* B-2020 ANTWERP *=* *=* Belgium *=* *=* tel: 32-3-2180247 *=* *=* fax: 32-3-2180257 *=* *=* e-mail: schryver-at-ruca.ua.ac.be *=* *=* homepage: http://www.ruca.ua.ac.be/EMAT *=* *=* *=* *=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*=*
One- or two-year POST-DOC position available at the Electron = Microscopy for Materials Science (EMAT) group of the University of = Antwerp, RUCA, Belgium. Do visit our website for more info on the = working of the group (http://www.ruca.ua.ac.be/EMAT).
The position is part of a joint project between EMAT and the = photographic company Agfa-Gevaert (3/99 - 3/01) on new photographic = materials and is open to inhabitants of member states of the European = Community or associated states. The candidate should have a large = experience in different TEM techniques for the characterisation of = atomic and microstructures. The position is immediately available. = Salary starts at =B1 1.500 Euro (after tax deduction), depending on = experience.
If you are interested or like more information, please contact Dr. D. = Schryvers (tel.: 32-3-2180247, fax: 32-3-2180257, e-mail: = schryver-at-ruca.ua.ac.be)
!!!!! NEW SERVER !!!!
DO CHECK MY E-MAIL ADDRESS: schryver-at-ruca.ua.ac.be (replies to old = mails, e.g. from before March 15, will not work anymore!)
*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*= =3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D* *=3D* *=3D* *=3D* Dr. D. Schryvers *=3D* *=3D* Electron Microscopy for Materials Research (EMAT) *=3D* *=3D* University of Antwerp, RUCA *=3D* *=3D* Groenenborgerlaan 171 *=3D* *=3D* B-2020 ANTWERP *=3D* *=3D* Belgium *=3D* *=3D* tel: 32-3-2180247 *=3D* *=3D* fax: 32-3-2180257 *=3D* *=3D* e-mail: schryver-at-ruca.ua.ac.be *=3D* *=3D* homepage: http://www.ruca.ua.ac.be/EMAT *=3D* *=3D* *=3D* *=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*= =3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*=3D*
I have just started trying a different approach on our intranet. The intranet runs on Microsoft BackOffice 2.5 and includes Exchange Server 4.0. I have setup mailboxes for our HRTEM and Ion Mill and hope to make use of the group scheduling and calendaring (is that a word?) functions of Exchange to achieve the same end. Client side software will be Exchange client / Outlook 97. Users can book by sending appointment requests to the mailboxes which can be set to automatically accept / suggest free time slots. I know this has access restricted only to our intranet unlike web based programs, but then at present we do not have scientists outside the organization directly booking resources. At present I am trying to get over the problem of compatibility: exchange client users do not have access to the outlook calendar and outlook users have chosen not to use schedule 97 as default calendar. If there are others using this method, I would like to hear from them as well as about web based programs.
--- R Divakar PMS, IGCAR, Kalpakkam 603102, India ----
This is a repeat of an earlier message that might not have made it. ;)
I have three (3) Osram UV bulbs that were used in an older, large, stand-alone UV lamp (The box is labeled: OSRAM Quecksilber-Hochstdruck-Lampe HBO 200W) If you need more info, or have need of these contact me direct. Thanks, john
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu
We have a system that we have just implemented (Actually my colleague=20 Corinna Wauchope did it) that uses Filemaker Pro to book instrument=20 time, track our user information, keep the instrument logbooks and=20 bill our users. It is working quite well now. We plan to put it on=20 the Web when we have all the glitches ironed out and all the=20 instruments included on it and whenCorinna has some spare time to do=20 the necessary Web stuff ( :-) ).
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Please note new FAX number.
John Mansfield PhD CPhys MInstP North Campus Electron Microbeam Analysis Laboratory 417 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (734) 936-3352 FAX (734) 763-2282 Cellular Phone: (734) 358-7555 (Leaving a phone message at 936-3352 is preferable to 358-7555) Email: jfmjfm-at-engin.umich.edu URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html Location: Lat. 42=B0 16' 48" Long. 83=B0 43' 48"
I run an EM lab at a small private college using a 1968 Hitachi Hu 125E and an old 1970 ETEC autoscan. !00% biological TEM & SEM, very few beam hours per year, no fees, no hassel. I've got spares in the closet, do 100% of my own service ~ with some help from my friends (THANKS to Allen Sampson).
Currently, I have said no to the question "buy a new scope", because what we have now has been working for 10 years, plus the money is not there. I feel that one is tied into a service contract with the purchase of a new instrument because of the complexity. If a new scope were purchased, no longer can you fix it with a relay, vacuum tube, or spare parts in the closet. Someday I will change my mind as the avaliability of parts dwindles and frustration levels go up.
I know our situation will not go on forever, and feel that planning for a new scope, service contract and in-house support should take place long before the old scopes become more trouble than they are worth. Your facutly members will have to be the ones to write grants to NSF programs and private foundations. Within the grant writing process, the questions that you ask will get answered as support either builds or dies for new instrumentation.
Robert
} } OK, I know asking this question of the group may seem strange, but this is } the most densely populated area of microscopy wisdom in the universe and I } need your global perspective. } } I am the only person in a small EM lab that serves a primarily } undergraduate campus. I am part of the technical staff, I am not a faculty } member. We have doctorate programs and a few researchers using TEM, but no } materials science types and no professional schools. I guess our TEM might } be used 300 to 500 hrs/yr sometimes less, never more. } } We have a JEOL 100B circa 1975. For most of our applications, it works } fine. I have a whole other microscope to use for parts and a clever service } provider who may be able to keep this thing going for a long time. In a } darkened room, it is hard to tell it from a more modern instrument based on } the images seen on the screen. When the lights go on, it looks 24 years } old. } } Recently a new faculty member asked when are we getting a new TEM, one with } digital imaging. I choked, said I would check into it. So here are a few } questions I need help with and I hope for some kindness and understanding } in your replies. I do need your replies because as the only real EM person } here I could use your help. } } I have never gone out looking for $$ for a new microscope. Where does one } look? Could we justify $$ based on our level of usage? What do you think? } What are the criteria used by funding agencies when evaluating requests. } Could you share you experiences? } } Suppose we can't get the $$ for a whole new TEM. Would I be crazy to try to } get $$ to equip our current TEM (remember 24 years old) with an add on } digital camera? The ports are there and it could be done, but I'm not sure } it is a good idea. } } I hope you can give me some leads and encouragement. Most of the burden of } doing this job will fall on me. Although often supportive in casual } discussions, I don't expect much help from anyone else here. } } Thanks } } Jonathan Krupp } Microscopy & Imaging Lab } University of California } Santa Cruz, CA 95064 } (831) 459-2477 } jmkrupp-at-cats.ucsc.edu
Robert Fitton Luther College Department of Biology 700 College Drive Decorah, IA 52101
Voice 319-387-1559 FAX 319-387-1080
Enjoy a visit to our website: http://www.luther.edu/dept/bio.htm
There have been several questions recently concerning the status of VCR Group. I have heard many rumors and a lot of talk "around the water cooler". I would like to dispel any rumors and clarify the situation. South Bay Technology and VCR Group have come to an agreement that will ensure the continuation of the VCR Group product line. The VCR products are being incorporated into the South Bay Technology line of products an= d will be manufactured and marketed by South Bay Technology, Inc. Althoug= h VCR Group, Inc. will no longer exist, that does not mean that the product= s will die or that support for existing customers will not be available.
We are in the process of transferring VCR operations and manufacturing to=
our corporate offices in San Clemente, CA. We expect to be fully operational by June 1 and will be able to fulfill orders shortly thereafter. Please be assured that we are here and available to support your VCR products and will be well into the future. In addition, to ensure a smooth transition, South Bay Technology has hired Vince Carlino, President of VCR Group.
As communications with VCR have been spotty over the past few months, I would encourage anyone with any questions about the VCR Group or products to contact me immediately with the relevant details. I will ensure that your order or problem is dealt with promptly=
and that your order is shipped within a realistic timeframe.
I thank you for this opportunity to set the record straight and I welcome=
all of our new VCR customers to the South Bay Technology family - I will look forward to hearing from you!
Best regards-
David Henriks Vice President
South Bay Technology, Inc. TEL: 800-728-2233 (tollfr= ee in the USA) 1120 Via Callejon +1-949-492-260= 0 San Clemente, CA 92673 USA FAX: +1-949-492-1499
Randi We use Trump's fixative for routine TEM analysis of surgical pathology specimens or surgical biopsies. Basically it is 4% formaldehyde and 1% glutaraldehyde in phosphate buffer. One reference available is Arch. Pathol Lab Med 100:405, 1976, this is a comparison of several fixatives, If I can help further, feel free to contact me. Marge
Margaret Springett e-mail hukee.margaret-at-mayo.edu IEM Specialist at Mayo Foundation 1426 Guggenheim Rochester, Mn. 55905
Steven, I'm sure you are aware there is a room temperature equivalent to your message, but I want to make sure that others on the Listserver don't conclude that cryosectioning is the only way to do this. For many materials, e.g. softer metals and some polymers, one can section smoothly at RT with a normal microtome and diamond knife. For many other polymers (and most biological materials), a smooth surface is produced at RT, but smearing distorts the shape of structural features. For harder crystalline materials, e.g. steels or intermetallics, a surface usually is produced which is microscopically rough to a degree dependent on the shear/brittle fracture details of the system, no matter what the temperature. Even more complex is the case for embedded ceramic/mineral particulate, fibers and the like, where the critical dimension seems to be size. Such features a few microns or less in diameter tend to section smoothly at either cryo or RT, but much larger, and they start to section with a rough surface (which still might furnish much useful information, of course).
I am encountering a growing number of microtomy workshop students, both private and public sector, who are using sectioning as a prelude to scanned probe studies, as well as optical, SEM and TEM. Thus the SPM community would do well to consider what is meant by a 'smooth' or 'rough' specimen surface in terms of what types of information can be collected realistically from one extreme to the other.
Tom Malis Group Leader - Characterization Materials Technology Laboratory Natural Resources Canada (Govt. of Canada) 568 Booth St., Ottawa, Canada ph. 613-992-2310 FAX 613-992-8735 email: malis-at-nrcan.gc.ca
} ---------- } From: Slap, Steven[SMTP:SSlap-at-ebsciences.com] } Sent: May 18, 1999 10:34 AM } To: Microscopy-at-Sparc5.Microscopy.Com } Subject: cryoplaning } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear fellow microscopists, } } I am looking for individuals interested in cryoplaning techniques (using } a cryoultramicrotome to produce blocks with a very flat surface and } examining this surface in a cryo-SEM). Please contact me back-channel } if interested. } } Best regards, } Steven E. Slap, Vice-President } ******************************************* } Energy Beam Sciences, Inc. } The Laboratory Microwave Company } Adding Brilliance to Your Vision } http://www.ebsciences.com {http://www.ebsciences.com} } ******************************************* } } }
Maybe someone has already suggested this to you, but if digital imaging is the only thing you lack, the cheap (but effective) way to go might be to invest in a good quality scanner. You scan in the negatives you shoot on your present instrument and you have instant digital images, plus an archival, high quality film backup in your originals. This will cost a fraction of adding on digital capabilities to your scope, especially if you already have a decent computer to run it from.
I am a big advocate of capturing images on film, then manipulating those images digitally. Highest quality, lower cost, and platform/software independent archiving of images. My two cents.
OK, I know asking this question of the group may seem strange, but this is the most densely populated area of microscopy wisdom in the universe and I need your global perspective.
I am the only person in a small EM lab that serves a primarily undergraduate campus. I am part of the technical staff, I am not a faculty member. We have doctorate programs and a few researchers using TEM, but no materials science types and no professional schools. I guess our TEM might be used 300 to 500 hrs/yr sometimes less, never more.
We have a JEOL 100B circa 1975. For most of our applications, it works fine. I have a whole other microscope to use for parts and a clever service provider who may be able to keep this thing going for a long time. In a darkened room, it is hard to tell it from a more modern instrument based on the images seen on the screen. When the lights go on, it looks 24 years old.
Recently a new faculty member asked when are we getting a new TEM, one with digital imaging. I choked, said I would check into it. So here are a few questions I need help with and I hope for some kindness and understanding in your replies. I do need your replies because as the only real EM person here I could use your help.
I have never gone out looking for $$ for a new microscope. Where does one look? Could we justify $$ based on our level of usage? What do you think? What are the criteria used by funding agencies when evaluating requests. Could you share you experiences?
Suppose we can't get the $$ for a whole new TEM. Would I be crazy to try to get $$ to equip our current TEM (remember 24 years old) with an add on digital camera? The ports are there and it could be done, but I'm not sure it is a good idea.
I hope you can give me some leads and encouragement. Most of the burden of doing this job will fall on me. Although often supportive in casual discussions, I don't expect much help from anyone else here.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Biological Measurements and Forces in MAC Mode™ AFM Stuart Lindsay, Wenhai Han, and Yanzhang Liu; ASU
Biological force measurements and the AFM Protein unfolding may play a vital role in protein function (Soteriou, Clarke et al. 1993). Last year, Rief et al. (Rief, Gautel et al. 1997) demonstrated that the AFM could be used to follow the unfolding of a single protein molecule trapped between the AFM probe and a gold surface
I have a colleague who would like to label antibodies for fluorescence for use in leaves, but would like an alternative to fluorescein. I believe Molecular Probes has quite a selection... are there some in particular that users have been happy with?
thanks in advance for your help shea
Dr. S. Shea Miller Agriculture and Agri-Food Canada Eastern Cereal and Oilseed Research Centre 2068 K.W. Neatby Bldg Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 email: millers-at-em.agr.ca phone: 613-759-1760 fax: 613-759-1701 ! !
} -----Original Message----- } From: Howard Fielding } Sent: Monday, May 17, 1999 1:25 PM } To: 'microscopy-at-msa.microscopy.com' } Subject: Electron Microscope For Sale } } Our firm has a ISI Electron Microscope, model DS 130 C for sale. Any } interested parties may call Howard Fielding at 800 992 7844 ext 304 for } information or respond via e-mail to hfielding-at-mayflower-sac.com. We have } acquired this device from a warehouse customer who has defaulted on } storage charges and can make a very attractive price to prospective } purchasers.
Does anyone know if in situ's can be done on epoxy embedded sections? If yes, does the tissue have to be treated any differently during fixation and embedding?
Thanks for all the interest, but the bulbs have already found a good home. We are also looking to get rid of some vacuum tubes (assorted sizes and manufacturers). All free for the cost of shipping. Spring cleaning, y'know? john
******************************************** John P. Shields Center for Ultrastructural Research 151 Barrow Hall University of Georgia Athens, GA 30602-2403 (706)542-4080 jpshield-at-arches.uga.edu
I am interested in exploring the feasibility of jet propane freezing our samples to minimize artifacts from ice formation, but am having a hard time finding a facility that has a rig. Any contact pointers would be very much appreciated, as would any clues on techniques used to minimize phase separation caused by the formation of ice. We are currently plunging our samples (colloidal dispersions) in liquid propane, but the size of the ice artifacts are on the order of our particle size.
I have been watching this discussion closely as I too am working with an antiquated scope, a 1975 Philips 201. I work only with clinical specimens, almost no research, and primarily muscle, nerve and renal biopsies. The 201 is functioning quite well also due in part to a master "Mr. Fixit". We have even more in common. I have a 201 parts scope sitting right next to my primary scope. I have sold the pathologists I do EM for on digital imaging, but due to tight purse strings am not going to be able to do what I would like to do. So, now to my question. I have been looking into the scanning possibilities and would love all the input you all could give me. I am looking to scan the standard Kodak 4489 negatives. What scanners are good for this? What price range am I looking at? Thanks in advance for all your input.
Douglas C. Rennie Electron microscopy lab coordinator University of Nebraska Medical Center Omaha Nebraska (402) 559-7729
We have an immediate opening in our facility for specialist in materials characterization. Areas of application include SEM/EDS, mProbe and ESCA/XPS. Background in inorganic chemistry, AA & ICAP are also desirable.
Breim Engineering is a laboratory environment that applies these techniques to metallurgical, materials characterization and failure analysis. Reply to: dlpark1-at-briemeng.com
********************************************************************** Donald L. Parker Briem Engineering, Inc 4134 Rider Trail North Earth City, MO 63045 [314] 298-3773, direct ext #27 FAX [314] 298-7097 **********************************************************************
We have a Hitachi HU12A to give away. It is a complete system with manuals. It is located at the Health Science Campus of USC in Los Angeles, CA. For additional information please email me, and I will get back to you asap. Thanks
At 09:36 AM 5/21/99 , you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Agfa Arcus II does 600x1200 dpi and runs about $850. The UMAX Powerlook III does 1200x2400 dpi and runs about $1200. Both do a good job. Higher resolution is good and the UMAX has a bit higher D range.
I need to find a light source, either a mirror or an electric lamp for several monocular light microscopes (Leitz). What would be the cost? Is there a generic brand that would work? Thanks for any suggestions!
Barbara Plowman Univ. of the Pacific School of Dentistry email: Bplowman-at-uop.edu
We are planing to immuno-gold label the melanocytes of equine melanoma. If you know of any commercial or non-commercial source of specific antibodies against melanocytes (such as Ab against melanosomes) please let us know. We would really appreciate that.
Regards
-Majid Ghoddusi
........................................................................... ........................................................................... ...... Majid Ghoddusi, PhD Centre for Microscopy & Microanalysis and Division of Veterinary Pathobiology & Anatomy The University of Queensland Qld 4072 Australia
To all who have replied to my inquiry, I want to thank you for being open and candidly sharing your experiences and expertise with an amateur like myself. It has been extremely helpful in clarifying my concerns.
I continue to welcome any additional input or comments.
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Without throwing the cat too heartily amongst the pigeons I would value your considered comment on the virtues of LVSEM against ESEM in a multi-user mainly biological EM facility.
Obviously I have investgated the subject as thoroughly as I can but there is nothing like a touch of personal experience to enrich this information for us,
Please send your responses directly to me by return or to bruton-at-emu.unp.ac.za. Our website (still under construction) address is provided beliow if you would like to know a little more about us.
I look forward to hearing your views.
Tony Bruton Head, Centre for Electron Microscopy University of Natal, Pietermaritzburg Tel +27 (0)331 260 5155 Fax +27 (0)331 260 5776 website:http:www.nu.ac.za (departments} units) Email:bruton-at-emu.unp.ac.za postal address; Private Bag X01, Scottsville, 3209 KwaZulu-Natal South Africa
Hi All: I am looking for a third party company that can offer me a service contract on a Philips 430 ST. Thank you so much for your ongoing assistance.
I recently bought a new (now discontinued) Wild M7S stereo Microscope, which included as part of the package a Wild model 10446174 trinocular phototube - this is the latest white version, 100% observation or 100% photo, built-in double iris diaphragm, 37mm photo port. This last part is driving me nuts, since all my photo equipment requires a 38mm photo port. Boring the phototube out to 38mm is not an option. What is an option is trading this to someone for an older black version with the 38mm photo port. The version that I have fits the M3 and M7, as well as all the current Leica stereo microscopes (MS5, MZ6, MZ8, etc.) - anything Wild or Leica except the M5 (this takes a different size, and they will not interchange). If you have a 38mm Wild phototube and are interested in becoming compatable with all of the new Leica digital and video adapters (which are 37mm, or so I understand), let me know and perhaps we can work out something.
Can anybody tell who are suppliers of gold-labelled Tumour Necrosis factor ? I don't seem to able to find anything in the catalogues I have at hand.
Thanks in advance
Cathy Gillespie
Cathy Gillespie Head Electron Microscopy/Histology/FACS Facility John Curtin School of Medical Research Australian National University Canberra Australia
Our Kevex 8000 system won't talk to an external computer and we would like to do peak to background analysis on spectra we have and will collect. It appears that Quantex will not give us straight P/B. Suggestions on how to proceed will be appreciated.
Dear Richard, I feel that it would be impossible to do in-situ hybridisation on conventional fixed and epoxy-embedded sections. There occur a few severe problems :
1.) Fixation: If you use conventional aldehyde-fixation, proteins as well= as nucleic acid will be crosslinked heavily and binding sites for your in-si= tu will be deleted
2.) Postfixation: Osmium will even increase the cross-linking and should = be avoided in any case
3.) Embedding: Using conventional hydrophobic epoxy resins you archive va= st three-dimensional cross linking of proteins, nucleic acids, amino-acids .= . Your in-situ probe will not have any access to binding sites.
If you have too much time/manpower or in situ hybridisation in absolutely necessary for your work you could try it with the following hints but I wouldn=B4t expect too much:
a) use a "soft" fixation (e.g. 2-4% Formaldehyd) b)don=B4t use osmium nor uranyl acetate c) don=B4t use epoxy resins, try more hydrophilic resins (e.g. LR-White o= r LR-Gold or the Lowicryls if you aren=B4t frightened of their toxic potential...) d) improve contrast of sections: strong postfixation and staining (why no= t including osmium)of sections after in-situ labelling
Another trial would be to try Ultracryotomy of cryo-protected deep frozen tissue (Tokaysu technique). But this will cost you a lot of equipment, ti= me and motivation ...
Good luck, with best regards Michael Reiner
Michael Reiner Department of Anatomy University of Cologne Germany e-mail: a2811111-at-smail.uni-koeln.de
Richard Gardiner schrieb:
} -----------------------------------------------------------------------= - } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Co= m } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm= l } -----------------------------------------------------------------------. } } Does anyone know if in situ's can be done on epoxy embedded sections? } If yes, does the tissue have to be treated any differently during } fixation and embedding? } } Richard Gardiner
Thanks very much to all who responded to my query about Trump's fixative. Got what I needed to finish processing the samples on hand.
Learned a couple of interesting things in the process: apparently there are two versions of Trump's---the standard one with paraformaldehyde and gluteraldehyde, and another one using formalin. The latter is probably a histology fixative, I guess. The other neat thing to know is that several people described Trump's as an excellent storage fixative. One person even said they'd had specimens in Trump's for nine years, with no noticeable deterioration!! (Don't try this at home folks?)
Anyway, thanks again. As usual, the list came through.
Dear Dr. Gauler, We got copy of your message and we wonder why you did not get in touch with our representative or with the field engineer in charge of your area. Eventually you can get in touch directly with us visiting our site www.leo.de We should like to help you to solve your problem, and to do that we need some information. Please, let us know: How long ago have been made the last service on the penning gauge. What is the vacuum reached in the column when the separation valve is closed. Where did you get the filaments you are using. How long ago did you get them. It could be helpful to know the serial number of your EM 10.
It may be that you have a vacuum problem or that something is wrong with your wolfram filaments. The all data we ask are useful for us to know the exact configuration of your microscope as well as to rebuild its all technical history. We are waiting for your answer in order to solve your problem. Thanks in advance for your cooperation.
LEO COE-TEM / Marco Arienti Carl-Zeiss Strasse, 56 D-73446 Oberkochen
Greetings to the Microscopy list. As a relatively new member of the list, this is my first request for help. I would like to know if is possible to obtain image of dendritic cells detected by elettron microscope. Furthermore if you know an internet site whit some informations related to these cells. Thanking you all in advance,
I would appreciate advice on the best way to obtain grain size statistics from a TEM image. Is there any software that will do this? Since we will be starting with a print from a TEM negative, are there any special scanner capabilities that are required? Any other suggestions on this process would also be appreciated.
Thank you for taking time to consider this request.
Sincerely,
Mick Thomas --------------------------------
Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
-----Original Message----- } From: Fagerland,Jane [mailto:jane.a.fagerland-at-abbott.com] Sent: Wednesday, May 19, 1999 5:59 PM To: Microscopy-at-sparc5.microscopy.com Cc: Ulrich,Roger
The Department of Microscopy and Microanalysis at Abbott Laboratories is recruiting a senior level microscopist for its Materials Science group.
Requirements include a Ph.D. in Materials Science, Chemistry, or related field, and a working knowledge of most of the following instrumentation: TEM, SEM, EDXS, XPS/SIMS, AFM, and light microscopy methods (polarized light, fluorescence, and confocal microscopy). Familiarity with spectroscopic techniques such as FTIR and a strong background in physical and analytical chemistry are desirable. Experience with biological systems would be highly advantageous.
We are looking for a team player with excellent interpersonal skills and the ability to readily adjust to rapidly changing priorities. The successful candidate will seek out and learn new technologies as needed to solve problems related to pharmaceutical and healthcare products. The ability to communicate clearly, both verbally and in writing, is essential.
Essential job functions: Independently design and carry out experiments to evaluate materials specimens by microscopic and microanalytical techniques. Develop methods using multidisciplinary approaches. Interpret data and effectively communicate results. Review data and reports for scientific integrity and clarity. Mentor career development of junior scientists. Present data at scientific meetings and publish in peer-reviewed journals. Provide technical and scientific leadership.
The Department of Microscopy and Microanalysis provides corporate-wide support in materials and biological microscopy to all Abbott Laboratories divisions. The facility houses two TEMs (a Philips CM12 STEM and a LEO 910), two SEMs (a Philips XL30 and AMRAY 1830i), three EDXS systems, a BioRad MRC 1024 UV confocal scanning laser microscope, a Digital Nanoscope III AFM with Bioscope, a Physical Electronics 5600 XPS/SIMS, and the usual assortment of light microscopes and sample preparatory equipment.
For further information, please contact:
Jane A. Fagerland, Ph.D. Abbott Laboratories D45M/AP31 200 Abbott Park Rd. Abbott Park IL 60064-6202
Be very, very careful concerning "software for grain size measurement from TEM images." Because grains seen in the TEM often contain dislocations, twins, stacking faults, high preferred orientation, etc; as well as having every shade of grey from B to W; ^ ANY ^ video-input/scanning based "automatic" grain size measuring machine will produce suspicious results. I didn't say they don't produce a "number," just that the "number" has little correlation with grain size. The human eye can easily see grains in a TEM image; machines can't.
What we do is measure the maximum diameter (chord) of about 200 grains in a series of TEM images using a digitizer. After entering the image's magnification into the PC connected to the digitizer, we 'click-on' each end of each grain's max chord. The PC calculates the chord length. When we think we have measured enough grains, we tell the PC to calculate the grain size. It calculates both the normal and lognormal grain sizes from the chord lengths (equations and correction factors from Shuckher's chapter in DeHoff & Rhines "Quantitative Microscopy"), tests which one better represents the measured distribution and tests whether we really have measured enough grains. Assuming it doesn't tell us to measure more grains, the computer plots a grain size histogram, overlays it with the calculated distribution for an eye-ball check, and reports all the normal and lognormal parameters (telling us which parameters are best). Besides using the human eye-brain for doing what it does best (judgement) and the computer for doing what it does best (crunching), the method as described beats automated systems time-wise (from walking in with an image to walking out with data), which we consider funny.
Having said all of this, I realise time marches on and things might have improved. If anyone out there has an automated machine that can measure grain size on my collection of images they are welcome try. They are: An 'easy' Mo grain size with no internal structure--just b/w grains; Cu grains with sharp b/w twins; ceramic grains with dislocation tangles and b/w gradation in the grains; An Al film with high preferred orientation such that neighbouring, similarly oriented, grains have nearly the same shade of grey; and a horrible duplex Cu distribution with a b/w gradiant across the image, twins, and dislocations.
Ron Anderson, IBM, Hopewell Jct., New York, USA. anderron-at-us.ibm.com
IBM Analytical Services; http://www.chips.ibm.com/services/asg
Mick wrote:
Fellow microscopists,
I would appreciate advice on the best way to obtain grain size statistics from a TEM image. Is there any software that will do this? Since we will be starting with a print from a TEM negative, are there any special scanner capabilities that are required? Any other suggestions on this process would also be appreciated.
Thank you for taking time to consider this request.
Sincerely,
Mick Thomas --------------------------------
Mick Thomas UHV-STEM Laboratory E-1 Clark Hall Cornell University Ithaca, NY 14853
If your 8000 has a printer, it is likely it will have (?) a 4 port serial card. The OSys "copy" command can be used to send a file out this port which can be received by a PC in the serial capture mode. Another option I have used is to equip a PC with a SCSI card and a drive like that used on the 8000 (I am now using Syquests, was using Bernoullii 44s previously). Using the program "RTCOPY" files saved in RT-11 format cam be copied into dos/windows on the pc.
EDS spectra can be saved as individual (ASCII - Lotus delineated) files using the command "SAV/EXT" and be exported using the above information. It will be up to Lotus / Excel or the like to do the P/B calculations. Alternately, you could save the raw spectrum, strip the background, then save the background subtracted version. the difference between them would yield the bkgn.
If memory serves: A simple alternate approach, if you only have 4 elements (8 windows max), is to paint "windows" (like selecting for external dot map) on the elements and adjacent background area. Save the window file, then recall/print windows. Intergals contained in each window should be printed.
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Our Kevex 8000 system won't talk to an external computer and we would like to do peak to background analysis on spectra we have and will collect. It appears that Quantex will not give us straight P/B. Suggestions on how to proceed will be appreciated.
by mail.mdacc.tmc.edu (8.8.5/8.8.5) with SMTP id LAA00748 for {Microscopy-at-MSA.Microscopy.Com} ; Tue, 25 May 1999 11:46:25 -0500 (CDT) Received: by utm-notes-m2.mdacc.tmc.edu(Lotus SMTP MTA v1.2 (600.1 3-26-1998)) id 8625677C.005D1064 ; Tue, 25 May 1999 11:56:30 -0500 X-Lotus-FromDomain: MDACC To: Microscopy-at-Sparc5.Microscopy.Com Message-ID: {8625677C.005C19C7.00-at-utm-notes-m2.mdacc.tmc.edu}
Douglas
I am currently getting good results wioth an older HP ScanJet 4c with a transparency adaptor attached to it. Unfortunatly, the transparency adaptor cost more than the Scanjet did in the first place.
Both of these pieces are over 3 years old and I am sure that there are newer, better and cheeper scanners with transparency adaptors on the market today.
Mannie steglich Tech Dir Path E M Lab M D Anderson Cancer Center
Does anyone out there know of an epoxy for TEM use that does not soften substantially at temperatures up to 500C? We want to do a high-temperature straining experiment but don't have too much material and would like to use a 3mm disk as the gauge with bits glued to the end for the 'grips'. Also, it would need to cure at less than 150C so we don't alter the microstructure before we start the test.
Thanks in advance.
Cheers, JSV *************************** John S. Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
We've just started acquiring digital 16 bit x-ray radiographs.
1) Is it worth doing 16 bit radiographs or will 8 bit do? 2) The images are HUGE! Also, much of our processing software only works well with 8 bit. Would converting from 16 to 8 bit be acceptable? If you don't for x-ray, please give your opinion for normal microscopy work.
Dear Jacob, We have done this on our Kevex 8000 and we just marked Windows of equal size on the peak and background regions and then read off the Integral of the Window (lower right-hand of the screen) after the count and copied it down manually. Clumsy, but it worked. You wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
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-interchg.ubc.ca
I agree with Ron, that grain measurements on TEM images are very hard to do and one should not trust fully automatic measurements just because they are done by a computer. The reasons are the contrast mechanisms in a TEM. Bend conours, orientation contrast, dislocations and other effects can lead to artifacts that can easily be interpreted as grain boundaries or confuse software with grains of different intensity levels.
However, we have developed software to analyze grains in SEMs, where we also have to deal with grains of different intensity. While I cannot promise that it will work on your TEM images in an automatic fashion, I would like to offer you a trial. Send me a couple of your images and I will try if the software can be used to measure the grain sizes. The software does intercept techniques, similar to what Ron describes, as well as fully planimetric measurements. It also offers a semiautomatic mode for grain boundary reconstruction, which might be useful in this case.
Again, send me those images and I'll give it a try.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
} ---------- } From: } "anderron-at-us.ibm.com"-at-sparc5.microscopy.com[SMTP:"anderron-at-us.ibm.com" } -at-sparc5.microscopy.com] } Sent: Tuesday, May 25, 1999 9:22 AM } To: microscopy-at-sparc5.microscopy.com } Subject: Re: Software for grain size statistics } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Mick, } } Be very, very careful concerning "software for grain size measurement } from TEM } images." Because grains seen in the TEM often contain dislocations, } twins, } stacking faults, high preferred orientation, etc; as well as having } every shade } of grey from B to W; ^ ANY ^ video-input/scanning based "automatic" } grain size } measuring machine will produce suspicious results. I didn't say they } don't } produce a "number," just that the "number" has little correlation with } grain } size. The human eye can easily see grains in a TEM image; machines } can't. } } What we do is measure the maximum diameter (chord) of about 200 grains } in a } series of TEM images using a digitizer. After entering the image's } magnification into the PC connected to the digitizer, we 'click-on' } each end of } each grain's max chord. The PC calculates the chord length. When we } think we } have measured enough grains, we tell the PC to calculate the grain } size. It } calculates both the normal and lognormal grain sizes from the chord } lengths } (equations and correction factors from Shuckher's chapter in DeHoff & } Rhines } "Quantitative Microscopy"), tests which one better represents the } measured } distribution and tests whether we really have measured enough grains. } Assuming } it doesn't tell us to measure more grains, the computer plots a } grain size } histogram, overlays it with the calculated distribution for an } eye-ball check, } and reports all the normal and lognormal parameters (telling us which } parameters } are best). Besides using the human eye-brain for doing what it does } best } (judgement) and the computer for doing what it does best (crunching), } the method } as described beats automated systems time-wise (from walking in with } an image to } walking out with data), which we consider funny. } } Having said all of this, I realise time marches on and things might } have } improved. If anyone out there has an automated machine that can } measure grain } size on my collection of images they are welcome try. They are: An } 'easy' Mo } grain size with no internal structure--just b/w grains; Cu grains with } sharp b/w } twins; ceramic grains with dislocation tangles and b/w gradation in } the grains; } An Al film with high preferred orientation such that neighbouring, } similarly } oriented, grains have nearly the same shade of grey; and a horrible } duplex Cu } distribution with a b/w gradiant across the image, twins, and } dislocations. } } } Ron Anderson, IBM, Hopewell Jct., New York, USA. } anderron-at-us.ibm.com } } IBM Analytical Services; http://www.chips.ibm.com/services/asg } } } Mick wrote: } } Fellow microscopists, } } I would appreciate advice on the best way to obtain grain size } statistics } from a TEM image. Is there any software that will do this? Since we } will } be starting with a print from a TEM negative, are there any special } scanner } capabilities that are required? Any other suggestions on this process } would also be appreciated. } } Thank you for taking time to consider this request. } } Sincerely, } } Mick Thomas } -------------------------------- } } Mick Thomas } UHV-STEM Laboratory } E-1 Clark Hall } Cornell University } Ithaca, NY 14853 } } Phone: 607-255-0650 } Fax: 607-255-7658 } e-mail: mgt3-at-msc.cornell.edu } } } } }
I believe I have the required software which can DO automatic GRAIN SIZING! (let us see the response to that... :)
However can it do exactly what you what have in mind ALL the time and automatically?
For that I would reserve judgment until I have seen an image what you are attempting to qualify.
Mr. Anderson has brought up a number of points that need to be kept in mind. I especially like the selective method of analysis - with the computer being used where it helped and not where it could/would not (be it time or method dependent).
} If anyone out there has an (should I add SEMI-? until I have seen the images) automated } machine that can measure grain size on my collection of images they are welcome try.
I accept your challenge and raise you =85.
Possibly the best way forward would be for both of you to send me "typical" images (of the best possible quality) and what you would like to measure. I can then see if the software can do what is required and send you (and who ever else may be interested) the results.
Barry T. Dudley DUDLEY-at-I-CUBEinc.com
PS - The software I have in mind is called Vision Gauge.
-- ************************************************************ B.T. DUDLEY I-CUBE http://www.i-cubeinc.com Ph 1-888-77-I-CUBE 301-858-0505 301-858-0615 (Fax) I-CUBE is a Systems Integrator and Value Added Reseller of image analysis and image processing products for scientific and industrial applications. We provide a single source for imaging products, using a consultative selling approach I-CUBE 2411 Crofton Lane; Suite 14A; Crofton; Maryland; 21114 ************************************************************
It is not an epoxy but we have used `Autostick' a high temperature adhesive (up to 1100 C) which will stick to keyless surfaces, eg. polished foils. I don't have the details to hand but if you need them let me know and I'll try to hunt them down.
Ron
} Does anyone out there know of an epoxy for TEM use that does not soften } substantially at temperatures up to 500C? We want to do a high-temperature } straining experiment but don't have too much material and would like to use a } 3mm disk as the gauge with bits glued to the end for the 'grips'. Also, it } would need to cure at less than 150C so we don't alter the microstructure before } we start the test.
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
} } To: Microscopy-at-Sparc5.Microscopy.Com } } Subject: TEM Availability? } } Content-Type: text/plain; charset=us-ascii } } Content-Transfer-Encoding: 7bit } } } } } I would appreciate information on availability of a used CM12/CM200 } } electron } } microscope. The ideal "candidate" would be equipped with a STEM unit, } } EDS, } } PEELS, CCD camera and located, preferably, in the U.S. Individual } } attachments listed would be considered as well. Please contact me off } } line } } if you can provide assistance in locating this equipment. } } } } Bob Roberts } } EM Lab Services, Inc. } } Tempe, Az 85282 } } } } }
I was visiting a colleague of mine and we discovered a cache of "obsolescent" computer goodies:
AMDEK Laserdrive 1 Hayes Smartmodem 2400 Spin Rite 3.1 (some kind of disk repair utility for PC) WordPerfect 5.1 upgrade for PC 20 boxes or so of 5.25 inch disks, most unopened, some disk boxes, etc.
Anyone want this stuff? I'd hate to pitch it if someone can use it. Contact me via e-mail and I'll despatch it appropriately. If more than one person wants it I'll divvy it up somehow...
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Fellow Microscopists, We are presently measuring particle size and position by LM on small particles dispersed over a large area. This is accomplished by a frame by frame raster. We are in the process of upgrading our system to accomplish this. My question concerns the possibility of using a linear CCD array and scanning the sample under the array to cover our area of interest. This would involve specialized software to interact with the stage control and some dynamic particle identification scheme. Has anyone out there in microland attempted any similar procedure. Russ Gillmeister Xerox
Emitech make three types of cold stages two of these are also preparation stages. The stages suit most, if not all SEMs. You can read up on these in our online on page E10D. We are making additon to this page and more complete info will be up within a week. Please note that ProSciTech distribute these instruments in Australasia (south of Singapore) only. For agents elsewhere see Emitech's email contact on page E10. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
-----Original Message----- } From: Marisa Ahmad [SMTP:mahmad-at-semiconductor.com] Sent: Wednesday, 26 May 1999 01:30 To: 'MSA listserver'
Looking for parts for a Zeiss OpMi-1 stereomicroscope. Please contact me off-list. Thank you.
James Martin Director of Analytical Services & Research Williamstown Art Conservation Center http://members.tripod.com/~James_Martin/dasrhome.htm
Research Scientist in Chemistry Williams College (www.williams.edu) http://members.tripod.com/~James_Martin
I have been using FITC-dextran as a permeability marker in rabbit aorta. I have tried using molecular weights of 10,000 and 70,000 separately. The tissue was immersion fixed in glutaraldehyde and post-fixed in 1% potassium ferrocyanate mixed with osmium tetroxide. The result was disappointing. I have not been able to find any dextran-ferrocyanate deposits between any gap junctions nor glycogen. Would anyone have any suggestions what else I can try? I have used horse radish peroxidase and know that it worked but cost was a major issue. Thank you for any help offered.
Fanny Chu
Mark Elliott, PhD Research Associate UBC-Pulmonary Research Lab St. Paul's Hospital Vancouver, BC Canada V6Z 1Y6 604-631-5351 (FAX)
The Science Illustration program here needs help learning about the features available and criteria to use when selecting a set of stereo microscopes for their students. They are about to purchase 10 scopes to be used by graduate level science illustration students. They think they need drawing tubes/camera lucida systems and want to provide their students with the opportunity to have drawing experience using a 'real' microscope.
So, all you Rembrandts and Picasso's out there, here is your chance to help out some talented and devoted artists. Send me your advice and I will forward it to them. Help them make an informed decision that will keep them a fan of microscopy.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Remember the size of the files in bytes will be the x-size times the y-size times 1 for 8-bit images and 2 for 16-bit images. So by going to 8-bit images you could cut your file size in half. However, more gains might be made by reducing your x and y resolution. (You did not mention what resolution you are using.) How much do you really need? Images over 1024x1024 are not very useful unless you are digitally magnifying them to see the details.
16-bit imaging is helpful if you have a wide range of gray scale. You can concentrate on bright areas of the image and adjust the brightness and contrast locally to see the details you want without having the gray level steps showing up. At 16-bit (256 gray levels) you will almost certainly see the steps after much less contrast enhancement.
You might want to keep your original image "as is" and pull out smaller regions that can be enhanced and save those enhanced files as 1024x1024 8-bit images. I venture to guess that the aggregate size of those "region of interest" images would be much less than your original.
At 01:39 PM 5/25/1999 -0500, you wrote: } We've just started acquiring digital 16 bit x-ray radiographs. } } 1) Is it worth doing 16 bit radiographs or will 8 bit do? } 2) The images are HUGE! Also, much of our processing software only works } well with 8 bit. Would converting from 16 to 8 bit be acceptable? If you } don't for x-ray, please give your opinion for normal microscopy work.
I am looking for eyepiece reticles/graticles for a Nikon SMZ-10 dissecting scope. In my catalogs I can find the most common diameters (19 and 21 mm), which are too small for my eyepiece, which measures about 24 mm inner diameter.
thanks in advance
Steve
--------------------------------------------------------------------- Dr. Steven Barlow, Associate Director EM Facility/Biology Department San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619)594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu website: http://www.sci.sdsu.edu/emfacility/
Chairman, Educational Outreach subcommittee promoting access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
Dear Microscopists, We would like to get a UPS for our TEM to bridge the gap between the instant there is a loss of power and the time when the backup generator takes over (a minute or so??). We have some information about APC and Tripp Lite UPS units, but we are concerned that their experience is limited to computer applications. We would like information on the types of UPS employed by EM users. We would also appreciate suggestions on how to chose the appropriate size. The power consumption of our TEM is about 3 kilowatts (instuction manual). Many Thanks in advance. Vachik Hacopian
I am pretty sure what you have in mind can be done, but what advantages do you think this would provide? For example, we have software that interfaces with stage controllers and allows you to do exactly what you want without having to develop new software. We do so by controlling the stage, set up a raster for image acquisition, then automatically move the stage, acquire an image, montage the images and do the analysis on them. Alternatively you can evaluate the image separately. For accurate statistics you need to set up the acquisition withput overlap in the latter case.
Unless there is an advantage for using a linescan CCD array (is there?) I would consider using off-the-shelf parts. Of course, I have a vested interest in this, as we are selling them, but I am sure there are other vendors out there as well.
Michael
Michael Bode, Ph.D. Soft Imaging System Corp. 1675 Carr St., #105N Lakewood, CO 80215 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
******************************************************* Disclaimer: Soft Imaging System produces and sells image acquisition and processing systems. We therefore have a vested interest in some of the items mentioned above. *******************************************************
} ---------- } From: Gillmeister, Russ[SMTP:RGillmeister-at-sdms.usa.xerox.com] } Sent: Wednesday, May 26, 1999 7:37 AM } To: 'MSA' } Subject: Particle Analysis } } ---------------------------------------------------------------------- } -- } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ---------------------------------------------------------------------- } -. } } } Fellow Microscopists, } We are presently measuring particle size and position by LM on small } particles dispersed over a large area. This is accomplished by a frame } by } frame raster. We are in the process of upgrading our system to } accomplish } this. My question concerns the possibility of using a linear CCD } array and } scanning the sample under the array to cover our area of interest. } This } would involve specialized software to interact with the stage control } and } some dynamic particle identification scheme. Has anyone out there in } microland attempted any similar procedure. } Russ Gillmeister } Xerox }
The graticule usually sits losely on a ledge within the ocular. So one mm less than the inner tube diameter is acceptable, three appears a bit much. Graticules are frequently made to order and the non standard sizes just cost a little more. Our info is online on page S3. I am happy to acknowledge that other suppliers likewise would be able to procure other sizes. Cheers Jim Darley
ProSciTech Microscopy PLUS PO Box 111, Thuringowa QLD 4817 Australia Phone +61 7 4774 0370 Fax: +61 7 4789 2313 Great microscopy catalogue, 500 Links, MSDS, User Notes www.proscitech.com.au
-----Original Message----- } From: Steve Barlow [SMTP:sbarlow-at-sunstroke.sdsu.edu] Sent: Thursday, 27 May 1999 06:19 To: microscopy-at-sparc5.microscopy.com
Dear members,
I am working in KRISS(korea research institute of standards and science) and studying micoelectromechanical systems characterization by home-built photothermal microscopy and calibrated atomic force microscope. I am seeking to find infrared microscope( not FTIR) which views temperature differences of micromechanical systems specimen at roomtemperature with a resolution of micrometer. Also I am seeking infrared detector array which can be used to make infrared microscopy.
=09As there have been several enquiries for details of=20 high temperature adhesive I am posting details to the list=20 rather than direct replies.
I originally used Autostick some 25 years ago and now=20 discover that I am no longer able to obtain it. The last=20 time I needed any I used the `Sensor and heating cement'=20 from Oxford Instruments (Fax +44 1865 393333 - part no TGZ=20 0005 costs about =A36 for 50 gm pot). This seemed to be very=20 similar and was successful in my use of sitcking heating=20 elements onto a hot stage. It does dry in air but needs to be outgassed before use in=20 the microscope to prevent contamination.
=09I believe that the original manufacturer of=20 Autostick was Cotronics Corporation of Brooklyn, NY 11235.=20 They have advertised high temp resins (up to 700F) and=20 cements (up to 3000F) as late as 1992 but in pint or quart=20 quantities. They did not include Autostick in the catalogue=20 but had a larger range of cements for even higher=20 temperatures.
Regards, Ron
I have no financial interest in either company.
---------------------- Mr. R.C. Doole Department of Materials, University of Oxford. Parks Road, Oxford. OX1 3PH. UK. Phone +44 (0) 1865 273701 Fax +44 (0) 1865 283333 ron.doole-at-materials.ox.ac.uk
E. Fjeld Co., Inc. has been manufacturing custom stages for Amray, JEOL, = and Hitachi SEM's for over 20 years. I have purchased several of their custom stages or stage accessories over the last 15 years and have been delighte= d with the performance. They are located at 3 Executive Park Drive, North Billerica, MA and their telephone number is (978)667-1416. Mark Reynolds,= VP of manufacturing, is a good listener and very helpful guide through the design and manufacture of whatever stage or SEM add-on you may be interes= ted in.
Regards,
/richard
-----Original Message----- } From: Marisa Ahmad [mailto:mahmad-at-semiconductor.com] Sent: Tuesday, May 25, 1999 11:30 AM To: 'MSA listserver'
Who out there makes custom stages for SEM's (specifically JEOL)?=A0 Pleas= e contact me if you do, or know of anyone who does.=A0 Thank-you!
Robert, In situ cleaning is probably not a viable procedure. I would think if someone would build a jig for supporting the cantilever then Co2 cleaning may be a viable, off line solution. The greater concern may be how the tips may survive the thermal shock. Russ Gilmeister Xerox
-----Original Message----- } From: Co2clean-at-aol.com [mailto:Co2clean-at-aol.com] Sent: Thursday, May 27, 1999 8:22 AM To: spm-at-di.com
Hi,
You can try G-1 epoxy which is heat curing epoxy designed for high temperature application. Gatan produces this epoxy. The price in the UK is around =A370. Studies at GATAN indicate, a technical information of G-1 said, that G-1 bound interfaces remain intact in a TE= M hot stage at temperatures in excess of 1000 C.
Good Luck.
P Wang Department of Engineering Materials University of Sheffield Sir Robert Hadfield Building Mappin Street Sheffield S1 3JD UK Tel: 0114 222 5973(O) 0114 222 5519(L) Fax: 0114 222 5943 E-mail: p.wang-at-sheffield.ac.uk
we want to embedd a small sample of polysulphone, but metacrylate changes the sample. Epoxide resins are also not possible, because we want to dissolve away the resin after cutting. Are any resins or embedding materials known, which are practicable for our effort?
Kind regards
Rainer Ziel
------------------------------------------------------------- Dipl.-Phys. Rainer Ziel Acordis Research GmbH Obernburg ACR-O/RMG-EM 63784 Obernburg Germany
At 03:28 PM 5/27/99 +0100, you wrote: } } Hi, } } You can try G-1 epoxy which is heat curing epoxy designed for=20 } high temperature application. Gatan produces this epoxy.
Gatan *SELLS* the epoxy. It bears a strong resemblance to EpoTek 353ND. =20
The price in=20 } the UK is around =A370. Studies at GATAN indicate, a technical=20 } information of G-1 said, that G-1 bound interfaces remain intact in a TEM= =20 } hot stage at temperatures in=20 } excess of 1000 C.=20 } } Good Luck. } } P Wang } Department of Engineering Materials } University of Sheffield } Sir Robert Hadfield Building } Mappin Street } Sheffield S1 3JD } UK } Tel: 0114 222 5973(O) } 0114 222 5519(L) } Fax: 0114 222 5943 } E-mail: p.wang-at-sheffield.ac.uk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University=09 (614) 292-0674 http://web.ceof.ohio-state.edu Murphy's Law: "If anything can go wrong, it will." Commentary: "Murphy was an optimist."
I'm looking for a small wafer of Ge. Something in the order of 1/2 to 1" in diameter would do. I want to use it as a target and sputter it in my BalTec ion mill to make a Ge film. I don't really care how pure it is or if it is polycrystalline. Does anybody have a small piece that they could spare?
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
I assume that you tried Gatan's G1 (same as Epoxy Technologies EPO-TEK 353ND) If I remember right, Gatan rates this up to 500C, but I think that people have pushed it higher for heating experiments in the TEM.
Check out Fiore and Herring's paper in the first MRS TEM sample Prep series book (MRS vol 115, p125). They use Ceramabond 569 from Aremco.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center Guys Run Rd. (packages) P. O. Box 11472 (letters) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8161 (fax)
"The opinions expressed are those of Scott D. Walck and not of PPG Industries, Inc. nor of any PPG-associated companies."
---------- } From: Vetrano, John S To: 'Microscopy Listserver' -----------------------------------------------------------------------.
Hi all;
Does anyone out there know of an epoxy for TEM use that does not soften substantially at temperatures up to 500C? We want to do a high-temperature straining experiment but don't have too much material and would like to use a 3mm disk as the gauge with bits glued to the end for the 'grips'. Also, it would need to cure at less than 150C so we don't alter the microstructure before we start the test.
Thanks in advance.
Cheers, JSV *************************** John S. Vetrano Pacific Northwest National Laboratory MSIN P8-16 P.O. Box 999 Richland, WA 99352 Phone: (509)372-0724 Fax: (509)376-6308 Email: mailto:john.vetrano-at-pnl.gov
Does anyone know how I can contact Virtual Laboratories (distributor of Desktop Microscopist). Their web-site is gone, and their phone has been disconnected. I am looking for a Mac-based version of the Electron Diffraction Database to link with DM. Thanks in advance.
Jim Cole
Jim Cole Argonne National Laboratory-West PO Box 2528 Idaho Falls, ID 83403-2528 (208) 533-7165 Fax (208) 533-7863
Thank you for the proofreading, Woody. You are correct that I meant that at the lower 8-bit, 256-gray-level resolution that steps in brightness would begin to appear after relatively little contrast enhancement.
At 07:35 AM 5/27/1999 -0500, "White, Woody N" {Woody.N.White-at-mcdermott.com} wrote: } Hello Warren, } } Good answer, but could you have meant 16 bit is 65.5K and 8 bit is 256 } levels? } ..Or did I miss something? Woody } } SNIP } steps showing up. At 16-bit (256 gray levels) you will almost certainly see } the steps after much less contrast enhancement. } SNIP
Hi All: I am trying to digitize and capture images from my 845 JEOL SEM. Has anyone done this themselves? What issues do I need to worry about? What capture cards are best? I am looking to do this with off-the-shelf components. Thanks and regards,
} ---... } } Good answer, but could you have meant 16 bit is 65.5K and 8 bit is 256 } levels? ..Or did I miss something? Woody } } SNIP } steps showing up. At 16-bit (256 gray levels) you will almost } certainly see } the steps after much less contrast enhancement. } SNIP }
Computer displays are not capable of displaying 65.5 gray levels. You are manipilating 65.5k gray levels but the display hardware is still showing you only 8bit grays. This fact doesn't take anything away from the fact you manipulate 16bit grays with better control, resulting in less gray level posterization.
cheerios, shAf
{} /\ {\/} /\ {\/} /\ {\/} /\ cogito, ergo zZOooOM /\ {\/} /\ {\/} /\ {\/} /\ {} Michael Shaffer, R.A. - ICQ 210524 Geological Science's Electron Probe Facility - University of Oregon mshaf-at-darkwing.uoregon.edu - http://darkwing.uoregon.edu/~mshaf/
A major reason for using 16-bit grayscale images (or 12-bit ones from today's 36-bit color flatbed scanners for that matter) is for image processing before converting the images to 8 bits (256 grays) for printing. Applying a series of image processing operations such as background subtraction, filtering, and brightness/contrast adjustments to an 8-bit image causes posterization (loss of gray levels) seen as stairstepped contrast in the image. This problem arises from doing integer math starting with only 256 numbers. Processing images at 16 bits avoids these problems.
Several common image processing programs have 16-bit capabilities. Photoshop 5 is one. It allows tonal adjustments at 16 bits and can be extended with third-party plug-ins to allow filtering (e.g., unsharp masking) at 16 bits. After processing, you can easily convert to 8 bits. Another feature of Photoshop worth a microscopist's understanding is using adjustment layers to process 8-bit images with minimal data losses.
As to the suggestion to decrease file sizes by cutting the image resolution to 1024 x 1024 pixels, this is OK for final processed images but will eliminate the possibility of later magnifying the images to observe fine details. Saving files with a compressed format such as JPEG will greatly reduce file sizes without affecting the resolution or greatly affecting the grayscale information content. There may also be file compression methods applicable for 16-bit images, but I'm not familiar with any.
My suggestion would be to acquire and process for background leveling, sharpening and tonal correction at 16 bits, then convert to 8 bits and archive at full pixel resolution, possibly in a compressed format such as JPEG to save space.
---------- From: Warren E Straszheim Sent: Wednesday, May 26, 1999 12:12 PM To: Microscopy-at-Sparc5.Microscopy.Com Subject: Re: 8 bit versus 16 bit images
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Remember the size of the files in bytes will be the x-size times the y-size times 1 for 8-bit images and 2 for 16-bit images. So by going to 8-bit images you could cut your file size in half. However, more gains might be made by reducing your x and y resolution. (You did not mention what resolution you are using.) How much do you really need? Images over 1024x1024 are not very useful unless you are digitally magnifying them to see the details.
16-bit imaging is helpful if you have a wide range of gray scale. You can concentrate on bright areas of the image and adjust the brightness and contrast locally to see the details you want without having the gray level steps showing up. At 16-bit (256 gray levels) you will almost certainly see the steps after much less contrast enhancement.
You might want to keep your original image "as is" and pull out smaller regions that can be enhanced and save those enhanced files as 1024x1024 8-bit images. I venture to guess that the aggregate size of those "region of interest" images would be much less than your original.
At 01:39 PM 5/25/1999 -0500, you wrote: } We've just started acquiring digital 16 bit x-ray radiographs. } } 1) Is it worth doing 16 bit radiographs or will 8 bit do? } 2) The images are HUGE! Also, much of our processing software only works } well with 8 bit. Would converting from 16 to 8 bit be acceptable? If you } don't for x-ray, please give your opinion for normal microscopy work.
I need to colorize some SEM photos (digital). I have Adobe Photo Deluxe Home Version but I hear that CorrelDraw or Adobe Photo Shop might be better. What does everyone else use? If anyone has any special techniques to share I'd love to hear them.
I have a microscope lens for our Agema (recently bought out by FLIR) Thermovision 900 Infrared Camera with spatial resolution of 25 microns but a MFOV (Measurement Field of View) of 250 microns. In other words, although I can see 25 micron objects, the temperature measurement is from a much larger area. I'm not sure if there are any IR microscopes out there that are much better.
Also, Barnes used to make infrared microscopes. I believe they're now QFI - try http://www.quantumfocus.com/
I am working in KRISS(korea research institute of standards and science) and studying micoelectromechanical systems characterization by home-built photothermal microscopy and calibrated atomic force microscope. I am seeking to find infrared microscope( not FTIR) which views temperature differences of micromechanical systems specimen at roomtemperature with a resolution of micrometer. Also I am seeking infrared detector array which can be used to make infrared microscopy.
by mailhub.iastate.edu (8.9.3/8.9.3) with SMTP id SAA00465 for {Microscopy-at-msa.microscopy.com} ; Thu, 27 May 1999 18:02:39 -0500 (CDT) Message-Id: {4.1.19990527175423.00986e20-at-pop-2.iastate.edu} X-Sender: wesaia-at-pop-2.iastate.edu X-Mailer: QUALCOMM Windows Eudora Pro Version 4.1
Our JEOL 840A has been digitized for years. I think there are already connections inside to take active control of the beam. We ended up with another panel to sort out the control signals since we had a LeMont Scientific image analyzer, a Kevex Delta EDS, and a JEOL add-on imaging system all with the ability to take control of the beam.
Currently, the LeMont is gone and the Kevex has been replace by an IXRF Systems EDS box which we now use to digitize images. It works well.
We also have a Quartz PCI passive system down the hall on our Hitachi 2460N. It works real well but is a bit pricy. It should work just as well on the JEOL.
While you might be able to do the work yourself using off-the-shelf parts (or get a grad student to do it (or maybe you are a grad student)), you probably don't want to. I have done a fair amount of hardware and software work in my time, but I am losing my stomach for it as I get older. The system I might create probably would lack much compared to the commercial units available.
So much for my 2 cents worth. Hope it helped.
Warren
At 01:38 PM 5/27/1999 -0500, you wrote: } } Hi All: } I am trying to digitize and capture images from my 845 JEOL SEM. Has } anyone done this themselves? What issues do I need to worry about? } What capture cards are best? I am looking to do this with off-the-shelf } components. } Thanks and regards, } } Mike Coviello } UT Arlington }
This position is open for applicants who live or would like to live in Israel.
This position is for an engineer in the Transmission Electron Microscopy (TEM) group and involves all phases of TEM support. Responsibilities include planning analytical strategies, TEM specimen preparation, TEM imaging and microanalysis, report writing, interfacing with other engineers and customers, communicating results, and supervising workflow duties necessary to keep the lab running smoothly.
Applicants should be self-motivated and capable of working with minimal supervision. The successful candidate should have a Ph.D. or MS degree in Materials Science, Physics or equivalent, with university training in TEM imaging and diffraction theory and practice, EDX and EELS theory and practice, familiarity with microelectronics specimen preparation, and familiarity with microelectronics device operation and construction. Ability to work in a team oriented environment and good communication skills are critical.
Contact Information:
Mitzi L. Ryerson - F18 Materials Analysis Group Leader F18 Intel Electronics, LTD. email - mitzi.l.ryerson-at-intel.com outside Israel number - 011-972-7-666-6217 voicemail within Israel number - 07-666-6217 voicemail
This is an opportunity to assume significant responsibilities with Mineral Technologies, an international resource and technology-based organization that develops and produces performance enhancing mineral products.
You'll be involved in a diverse range of short/long-term projects and solve complex problems using electron microscopy and microchemical analysis of minerals and industrial products.
Duties involve providing/directing chemical and physical analyses of routine and complex samples: issuing oral/written technical reports of investigations, including creative conclusions and interpretations to solve technical problems. You'll also ensure that records are legal and
correct, and ensure a safe working environment that complies with EHS regulations; maintain continuous dialogue with customers and analytical services group members; actively support a total quality philosophy and help develop/implement a ISO Guide 25.
B.S. in Physical Science or Engineering required. Chemistry/Microscopy preferred. Must have at least 6 years of chemical analysis/microscopy, preferably in a service environment. Strong people, computer and oral/written skills essential.
We offer a competitive compensation and benefits package, opportunities to assume significant responsibilities and strong growth potential. For consideration, mail/fax resume with salary requirements to:
Gary Duckwall, HR Manager Minerals Technologies, Inc. 640 N. 13th Street Easton, PA 18042 Fax: 610-250-3210
Diane Ciaburri wrote: ================================================= I need to colorize some SEM photos (digital). I have Adobe Photo Deluxe Home Version but I hear that CorrelDraw or Adobe Photo Shop might be better. What does everyone else use? If anyone has any special techniques to share I'd love to hear them. ================================================== There is a product called "Spectrum Color" and you can find full details on it on the SPI Supplies website at URL http://www.2spi.com/catalog/software/spectrum-2.html
It is very low cost and very effective.
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: www.2spi.com ############################ ==================================================
Assuming that the internal hooks exist, as Warren notes, or can be obtained from the manufacturer, as in some Hitachi units, then a home grown device requires 2 channels of D/A and one of A/D. The D/As should be 16 bit devices though 14 is probably adequate. The A/D should be at least 8. Note that A/D speed is not a crucial variable since you can sit as long as you need to on a single point. (Though other dwell considerations obviously apply.)
Sorry if this seems to state the obvious. The hardware isn't the problem, it is the time to get it working satisfactorily.
Rich } } } Our JEOL 840A has been digitized for years. I think there are already } connections inside to take active control of the beam. We ended up with } another panel to sort out the control signals since we had a LeMont
} Dear all, } } we want to embedd a small sample of polysulphone, but metacrylate changes } the sample. Epoxide resins } are also not possible, because we want to dissolve away the resin after } cutting. Are any resins or embedding materials known, which are } practicable for our effort? } } Kind regards } } Rainer Ziel } } ------------------------------------------------------------- } Dipl.-Phys. Rainer Ziel } Acordis Research GmbH Obernburg } ACR-O/RMG-EM } 63784 Obernburg } Germany } } Tel: +49 (0) 6022 / 81-2645 } Fax: +49 (0) 6022 / 81-2896 } E-mail: Rainer.Ziel-at-AkzoNobel.com } } } } }
} Does anyone know how I can contact Virtual Laboratories (distributor of } Desktop } Microscopist). Their web-site is gone, and their phone has been } disconnected. I am looking } for a Mac-based version of the Electron Diffraction Database to link with } DM. } Thanks in advance. } } Jim Cole
Jim and all,
Maybe you tried the old web address at Rt66.com. They moved ISP and are now at http://www.easystreet.com/~lacuna/FrontPage.new/FrontPage.html It took me a bit of web searching to find this out for ourselves a few months ago.
By the way, maybe those who maintain lists of www addresses for microscopy companies (e.g. EMYP at Lausanne and Nestor's list at ANL) should update this link.
Anyway, I hope this helps.
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics Chalmers University of Technology S-412 96 G=F6teborg Sweden Tel: +46 31 772 36 33 FAX: +46 31 772 32 24 email: maclaren-at-fy.chalmers.se or: ianmaclaren-at-hotmail.com Research Group Homepage: http://fy.chalmers.se/microscopy/ +++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++
} } My suggestion would be to acquire and process for background leveling, } sharpening and tonal correction at 16 bits, then convert to 8 bits and archive } at full pixel resolution, possibly in a compressed format such as JPEG to save } space. } From: Warren E Straszheim
Once you have decimated the data either by lowering the resolution or the number of bits you can never get it back. CD ROM's are cheap so I save as much data as I can. It has proved very worth while if I have to go back and fix something later. For the same reason I always save the data in a raw format just as it comes from the source.
Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
} From: RCHIOVETTI-at-aol.com } Received: from RCHIOVETTI-at-aol.com (3950) } by imo23.mx.aol.com (IMOv20) id 8GTGa11104; } Wed, 26 May 1999 20:50:16 -0400 (EDT) } Message-ID: {85f34a05.247df0c7-at-aol.com} } Date: Wed, 26 May 1999 20:50:15 EDT } Subject: Re: eyepiece reticles/graticles } To: sbarlow-at-sunstroke.sdsu.edu, microscopy-at-sparc5.microscopy.com } MIME-Version: 1.0 } Content-Type: text/plain; charset="us-ascii" } Content-Transfer-Encoding: 7bit } X-Mailer: AOL 3.0 16-bit for Windows sub 38
} } Hi Steve, } } My favorite company for rulings, reticles, stage micrometers, etc. is as } follows: } } Klarmann Rulings, Inc. } P.O. Box 4795 } Manchester, NH 03108 } Tel. (800) 252-2401 } Fax (603) 424-0970 } http://www.reticles.com } } They have every kind of ruling and reticle imaginable, all sizes, custom } ones } as well, and if you need it, they can provide scales which are traceable to } NIST. } } Cheers, } } Bob } ****************************** } Robert (Bob) Chiovetti, Ph.D. } Microimaging Technologies, Inc. } Tucson, Arizona USA } Tel./Fax (520) 546-4986 } rchiovetti-at-aol.com } Manufacturers' Representatives } Systems Integrators } Analog & Digital Imaging Systems } Clinical & Research Microscopy } Cytology/Histology/Pathology/EM } *******************************
Hi. I would like to get 0.5% ruthenium tetroxide in ETHANOL for my freeze substition experiment. Does anybody know where I can get it? I appreciate if somebody can provide the information about where I can get ruthenium tetroxide solution (in ethanol) or ruthenium tetroxide crystal. Thanks.
At 03:29 PM 5/27/99 -0700, you wrote: } } Several common image processing programs have 16-bit capabilities. } Photoshop 5 } is one. It allows tonal adjustments at 16 bits and can be extended with } third-party plug-ins to allow filtering (e.g., unsharp masking) at 16 bits.
I certainly agree as to the necessity of keeping as much image depth as possible during the various processing steps. I would like to know what plug-ins will allow 16-bit manipulations? I've got John Russ's IP toolkit, but those only seem to work on 8-bit images.
} possibility of later magnifying the images to observe fine details. Saving } files with a compressed format such as JPEG will greatly reduce file sizes } without affecting the resolution or greatly affecting the grayscale information } content. There may also be file compression methods applicable for 16-bit
I would hesitate to use JPEG for pre-presentation images. Although the eye will often not see the degradation in JPEG images, it is definitely there. The various IP algorithms will sometimes enhance the JPEG artifacts. e.g. Try running an edge filter in a JPEG. You will see the squares that JPEG creates during the compression process ( I can send you a powerpoint slide illustrating this!)
That said, I often use JPEG compression after all my processing is done and I have my image set to its final size and resolution. Unless you enlarge the image, the eye won't see most of the JPEG artifacts.
Cheers, Henk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu "The wise are pleased when they discover truth, fools when they discover falsehood."
I have inherited an OLD International Equipment Co. IEC CTF microtome cryostat Model# AB4757, minus documentation and chucks. It is apparently in working order, but its previous users are no longer around. If anyone knows where I could obtain parts and a manual, please let me know. Thanks in advance. Regards,
Andrew Ochalski, Microscopy Technician, University of Ottawa Dept. of Biology, Room 108, Gendron Bldg. 613-562-5800 x6343
Here's an interesting note from kodak. Basically they are saying that for digital images one of the biggest advantages (now they are selling a film scanner here) of this scanner is that you can store (or archive) your images on FILM and then scan them only when you need them. So the big boys in digital imaging, are actually agreeing with some thing we as microscopists have been saying for awhile: The best archiving of images is actually on FILM (which we know has a life expectancy of over 165 years).
Taken from: http://www.kodak.com/cluster/global/en/service/faqs/faq1035.shtml
1.What is a film drive? The KODAK ADVANTIX Film Drive FD 300 is a desktop scanner designed to scan film in Advanced Photo System (APS) film cassettes.
The name "film drive" indicates a new technological concept that uses the film cassette as an image storage device. Rather than take valuable space storing image data on the computer's hard drive, Zip, Jazz drives, or floppy drives, the `film drive' scans APS film,DIGITIZES THE IMAGE FOR CURRENT WORK, AND THE IMAGE DATA REMAINS ON THE FILM IN THE CASSETTE.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu
Interestingly, we asked that question on our Cell Biology survey this past December. The results were a staggering 46 % of the total population (495 microscopists) said that they use Adobe Photoshop. When the data is analyzed in terms of the number of responses to that particular question (it was part of a special section on image analysis, so not everyone answered), the number jumps to a whopping 86%! Guess that answers your question.
Best regards, Barbara Foster Consortium President Microscopy/Microscopy Education ...Educating microscopists for greater productivity.
125 Paridon Street Suite 102 Springfield, MA 01118 PH: (413)746-6931 FX: (413)746-9311 email: mme-at-map.com Visit our web site {http://www.MME-Microscopy.com/education} ****************************************************** MME is America's first national consortium providing customized on-site workshops in all areas of microscopy, sample preparation, and image analysis.
At 04:52 PM 5/27/99 -0500, diane.a.ciaburri-at-gdds.com"-at-Sparc5.Microscopy.Com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Time is drawing near for finalizing the next issue of MSAs newsletter, "The MSA Bulletin" (the listserver would not let me use the title in the subject line, since it thinks it is apparently full of Bull). If you have anything that you would like to see included in this version of the MSA Newsletter, (and thereby prove the computer wrong) I need to have it in my possession by June 4th.
You can submit material to me electronically (preferred) or by hardcopy at the address shown below.
Thank you for your attention,
__________________ Stuart McKernan stuartm-at-tc.umn.edu Microscopy Specialist Office:(612) 626-7594 CIE Characterization Facility, University of Minnesota Desk: (612) 624-6009 100 Union Street S. E., Minneapolis, MN 55455 Lab: (612) 624-6590
} } } } } Please visit us at http://www.southbaytech.com { { { { {
Manufacturers of precision sample preparation equipment and supplies for metallography, crystallography and electron microscopy.
Message text written by "Ian MacLaren" } Jim and all,
Maybe you tried the old web address at Rt66.com. They moved ISP and are now at http://www.easystreet.com/~lacuna/FrontPage.new/FrontPage.html It took me a bit of web searching to find this out for ourselves a few months ago.
By the way, maybe those who maintain lists of www addresses for microscopy companies (e.g. EMYP at Lausanne and Nestor's list at ANL) should update this link.
Anyway, I hope this helps.
+++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++++ Ian MacLaren Department of Experimental Physics {
} ...Saving files with a compressed format such as JPEG...
JPEG is a very lossy algorithm. Even at a "high" quality setting, a good deal of information will be lost. Resolution will be noticeably reduced, and grayscale values will be changed enough that the image will be unsuitable for some modes of processing and many types of digital analysis. LZW compression, while not providing as small a file size as JPEG, is lossless, as are the algorithms applied to images in Photoshop's native format.
Thanks to all those who responded to my question on scanning EM negatives. UMAX seems to be most commonly used.
I would like to ask a few more questions.
1. A person here who has done some negative scanning found the biggest problem not to be resolution, but grey scale replication. Is reproduction likely to be similar for most scanners, or are some better than others?
2. The Agfa DuoScan has been suggested to us here, though no one mentioned it on the server. I wondered if anyone had experience with it.
3. Ideally, we would like something that can also do a good job on 35mm slides, is reasonably easy to use and is not too slow.--(not asking for much!). I know there are some nice 35mm scanners, but are flat bed scanners able to do both sizes well?
Hi All, We are looking for a sample holder capable of rotational and tilt movements, thats fits on a Zeiss 10 TEM. Would be very gratefull to hear from anyone who has one or has leads to a used or new one. Thank you.
The plug-ins I was referring to are the Deep Bit Filters from ImageExpress (http://www.scanprep.com/). You can get a trial copy of the filters from their webside.
Must admit I probably went too far recommending JPEG compression for archival images. I wouldn't want to mislead anyone about the losses involved. In Robin's case of 16-bit x-ray images, I could easily imagine 50MB or larger files that would "quickly" fill a CD.
Larry
---------- From: Hendrik O. Colijn Sent: Friday, May 28, 1999 6:30 AM To: Microscopy-at-Sparc5.Microscopy.Com; Thomas, Larry Subject: RE: 8 bit versus 16 bit images
Hi Larry,
At 03:29 PM 5/27/99 -0700, you wrote: } } Several common image processing programs have 16-bit capabilities. } Photoshop 5 } is one. It allows tonal adjustments at 16 bits and can be extended with } third-party plug-ins to allow filtering (e.g., unsharp masking) at 16 bits.
I certainly agree as to the necessity of keeping as much image depth as possible during the various processing steps. I would like to know what plug-ins will allow 16-bit manipulations? I've got John Russ's IP toolkit, but those only seem to work on 8-bit images.
} possibility of later magnifying the images to observe fine details. Saving } files with a compressed format such as JPEG will greatly reduce file sizes } without affecting the resolution or greatly affecting the grayscale information } content. There may also be file compression methods applicable for 16-bit
I would hesitate to use JPEG for pre-presentation images. Although the eye will often not see the degradation in JPEG images, it is definitely there. The various IP algorithms will sometimes enhance the JPEG artifacts. e.g. Try running an edge filter in a JPEG. You will see the squares that JPEG creates during the compression process ( I can send you a powerpoint slide illustrating this!)
That said, I often use JPEG compression after all my processing is done and I have my image set to its final size and resolution. Unless you enlarge the image, the eye won't see most of the JPEG artifacts.
Cheers, Henk
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://web.ceof.ohio-state.edu "The wise are pleased when they discover truth, fools when they discover falsehood."
Shanling Shi wrote: ================================================= Hi. I would like to get 0.5% ruthenium tetroxide in ETHANOL for my freeze substitution experiment. Does anybody know where I can get it? I appreciate if somebody can provide the information about where I can get ruthenium tetroxide solution (in ethanol) or ruthenium tetroxide crystal. Thanks. ================================================== RuO4 is quite explosive. Indeed a relatively small amount when it goes off can be quite devastating. One precious metals supplier (in the USA) about twenty five years ago, had some go off while in their storage facility and it caused quite a mess and a huge amount of damage.
Polysciences at one time offered it as a solution in chloroform, however I don't know if they are still doing that or not. It was quite unstable in this form as well, however.
Because of the extreme explosive hazard, there are major restrictions on shipping the material in solid crystals form. I have been out of touch with the shipping aspect of the problem so I can't not speak with authority on today's regulations. Even if it could be shipped,we ourselves would have to give it a careful look in terms of the potential liability exposure should some kind of accident occur.
We have had reports of users of our ruthenium tetroxide kits generating the tetroxide, and then collecting some of the vapors on a cold finger suspended over the aqueous solution. Once a given amount was collected, it was then immersed in carbon tetrachloride, warmed slightly, and the result was a carbon tet solution. I don't know about the solubility of RuO4 in ethanol, but if there was some solubility, then I presume an alcohol solution could be prepared in a similar way.
More information about ruthenium tetroxide and its dangers can be found on our website given below.
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: www.2spi.com ############################ ==================================================
} Must admit I probably went too far recommending JPEG compression for archival } images. I wouldn't want to mislead anyone about the losses involved. In } Robin's case of 16-bit x-ray images, I could easily imagine 50MB or larger } files that would "quickly" fill a CD. } } Larry
We don't know how long silver images will last at least 150 years or more. You can view them with out any hardware and you can always scan them in again when computer get more powerful. Nearly everybody is set up to handle them.
They do take up more space then digital but you can always get to them and they will always be compatible. A 4X5 negitive stores a lot more information than you can scan in.
IMHO Gordon
Gordon Couger gcouger-at-couger.com 624 Cheyenne Stillwater, OK 74075-1411 405 624-2855 GMT -6:00 www.couger.com/gcouger
I recently had to re-order Ruthenium powder and my less than exhaustive search and interview with an msds and the manufacturer found no mention or link to the explosive nature that I'd heard about here. The received the order by Fed X the next morning. It may be that this particular compound is not as unstable. Please help me by sending me a reference to this 'explosive risk'. The powder is very hydroscopic and must be kept desiccated, I believe. I'll look up the source and post next week. Jeff Day/'JD'
We have just started to use PhotoShop LE after previously using Adobe Photo Deluxe Home Version. It appears to be fairly easy to use.
J. Roy Nelson Material Testing Lab. Pennington, NJ
Hi all! I need to colorize some SEM photos (digital). I have Adobe Photo Deluxe Home Version but I hear that CorrelDraw or Adobe Photo Shop might be better. What does everyone else use? If anyone has any special techniques to share I'd love to hear them.
Thanks to all who answered my plead for MSmouse. That problem is resolved. The same computer (can not operate old video boards with PII or PIII processors!), is now really sick. It is a tower with magitronic BIOS and generic 486/66 mother board and Intel processor. The RAM sockets are defective and there may be a crack on the mother board. I need at least 3 ISA 16 bits slots and 2 8 bits. The slots should NOT extend over the microprocessor because the boards need sitting low and extend the entire length of the mother board. If anyone has information on how to obtain similar or equivalent mother board please let me know. Additionally, someone may have an old 486 like it collecting dust and I would like to have it. Call me if you do. Thanks.
*Disclaimer: Whatever... is not Tulane opinion! Cesar D. Fermin, Ph.D. Professor of Pathology & Otolaryngology web:[ http://www.tmc.tulane.edu/ferminlab/] or [http://www.tmc.tulane.edu/imaging/] Internet: [cfermin-at-mailhost.tcs.tulane.edu] 1430 Tulane Ave/SL79 New Orleans, La 70112-2699 Fax 504 587-7389, Voice 584-2521, Main Office 588-5224
Who built the first TEM in North America? Dr. Anderson of Washington State university built one in 1935, but I have heard that a university in Torono may have had one before WSU. Toronto is usually credited with the first microscope in 1939, but the microscope on display in the museum at WSU is dated 1935. Thanks, Will Garrett ssrs396-at-wsu.edu
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