Altera Corporation SEM/EDX Technologist (Reliability Technician Opportunity)
COMPANY: Altera Corporation is a leader in advanced programmable logic devices and software. Our continued growth and success necessitates the need for the following talented individual to join our team.
JOB DESCRIPTION: In this position you will work with product, reliability and technology engineers to identify appropriate failure analysis techniques, provide physical analysis service to identify the root causes of the failures,
document the results, and inform the engineers of the root causes for corrective actions. You will also coordinate the maintenance of failure analysis equipment.
QUALIFICATIONS: The ideal candidate will have one plus years related experience in physical analysis. Also they will be familiar with usage of analysis equipment including SEM, EDX, precision cross sectioning, RIE, Jet Etcher and chemical de-processing set-up. Good communication skills, ability to handle multiple projects and interact effectively with others is a must.
BENEFITS: Altera offers a challenging environment along with comprehensive benefits which include profit sharing, company matching 401(k) plan and an Employee Stock Purchase Plan.
CONTACT: Principals only please. EOE. Send resume to:
Altera Corporation, Human Resources, Attn: JG/SJDC295, 2610 Orchard Parkway, San Jose, CA 95134-2020. FAX: 408-435-5065.
} Diana van Driel wrote: } I was recommended to clean any gunked up lens with a knob of polystyrene } foam - no chemicals, just the dry foam rubbed in a circular motion onto the } lens. Over the years none of my Zeiss lenses has suffered any damage from } this.
Have you inspected the front lenses of your objectives under magnification to see if there are scratches in the coating? If there are none, is is a testament to the durability of the Zeiss coatings.
I have seen too many microscope lenses (yes, even Zeiss) marred by rubbing with dry materiels. I don't recommend this for any lens. The dust particles that settle on lenses, particularly uncovered oculars, act like sandpaper when rubbed across the lens surface. Solutions such as lens cleaners lubricate these particles and reduce their abrasive effect. Cotton buds tend to lift the particles into their fibers, while, I believe, polystyrene would tend to keep them at the lens surface where they can do damage.
This discussion of cleaning gunked immersion lenses misses the real problem, in my mind. I care for 2100 microscopes constantly in use by university students. We use only synthetic immersion oils. Despite warnings, a certain number of these microscopes end up with immersion oil on the high dry (40X) objectives, rendering them useless until cleaned. Since the microscopes may be of 8 different brands, of varied models and ages, my cleaning method has to work for them all, and be applicable in a classroom situation. Cotton buds and glass cleaner (Windex or something similar) used in a wash and dry pattern, work well for cleaning all lenses and is relatively benign to microscope and student. Glass cleaner doesn't work well for immersion oil on the 40X, because it requires repeated application of the glass cleaner to remove all traces, a time-consuming effort. The US Government, and particularly the state of California, have stringent safety rules governing use of chemicals. While xylene, may be the best cleaner for immersion oil, its toxicity is a drawback and I hesitate to recommend it for students in a classroom environment. Same goes for pet ether, methyl-ethyl ketone, acetone or chloroform. Alcohol would be a preferred solution, but, because of warnings about its use with older microscopes, I hesitate to recommend it. Perhaps when the older microscopes are completely phased out of our inventory ... Immersion oil seems to be a popular thread with this list. I would appreciate any comments or ideas.
Rick Markgraf Microscope Services University of California, Davis rlmarkgraf-at-ucdavis.edu PH. (916)752-3477 FAX (916)752-6363
There is an immediate opening in our group at Northwestern University for a Postdoctoral Scholar or a Research Associate, in the area of Analytical Electron Microscopy/Materials Science. I would greatly appreciate it if you could pass on the following information to interested parties. Please have anyone interested contact me at the address/e-mail/phone below.
POSITION OPEN: Postdoctoral Scholar or Research Associate
Area: Analytical Electron Microscopy & Materials Science
Qualification: A PhD in Materials Science/Physics or related area. Very strong hands-on experience in various AEM techniques is required, including transmission EELS, x-ray microanalysis and CBED. Experience in specimen preparation of variety of materials, especially x-sectional TEM is required. Preference will be given to those who have demonstrated skills and publications using FEG TEM/STEM/SEM and interfacial/defect phenomena in solids.
Instrumentation available at NU in the newly restructured Electron Probe Instrumentation Center (EPIC) includes: A Hitachi HF-2000 FEG TEM/STEM with x-ray, EELS, in-situ IV/LHe holder and e- holography set-up, a Hitachi S4500-II FEG SEM with x-ray, EBSP/OIM and LHe stage with in-situ IV probes, a Hitachi H8100 cTEM, and access to various other TEMs/SEMs and other analytical instrumentation.
Research involves use of diverse AEM techniques to probe problems of thin film growth, interfaces & grain boundaries in oxide systems.
The position is open immediately for at least one year, renewable upon mutual agreement for longer period. Salary and benefits will commensurate with experience and skills.
Please contact immediately:
******************************************************* (Vinayak P. Dravid) Associate Professor, Materials Science & Engineering Director, Electron Probe Instrumentation Center (EPIC) Northwestern University, Evanston, IL 60208, USA Ph.: (847) 467-1363, Fax: (847) 491-7820 E-mail: v-dravid-at-nwu.edu *******************************************************
Thanks,
(NOTE THE NEW AREA CODE) ******************************************************* (Vinayak P. Dravid) Associate Professor, Materials Science & Engineering Director, Electron Probe Instrumentation Center (EPIC) Northwestern University, Evanston, IL 60208, USA Ph.: (847) 467-1363, Fax: (847) 491-7820 E-mail: v-dravid-at-nwu.edu *******************************************************
I need help in either getting articles or article references on a good, short EM paper that would be useful for teaching students. In the July-August 1992 American Scientist Magazine, there was an excellent article, entitled "The New Vision of Light Microscopy". This is an excellent paper on LM and image analysis for students and also for technician references. If anyone out there has any suggestions or ideas, please let me know. Thank you in advance. Cathy Kelloes
In the past I have used chloroform routinely for expanding thin sections. Because of the hazardous nature of this chemical I tried a heat pen (amps - .125), but it was totally ineffective. Would a more powerful heat pen be effective in reducing compression ? I would appreciate some feed back on this. Thanks Leo
While discussion is going on about the precipitate that is forming on grids even when ultrapure boiled water is used (Cartwright, Lovett). I have another question. I see sometimes grids jumping in plastic dish becose of static electricity. I imagine that if we place charged grid on stain solution that some stain will precipitate.Is out there anybody who would comment for this possibility? John Gabrovsek
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At 04:28 PM 2/1/96 -0600, you wrote:
"Do you wet your grids before putting them in your methanolic uranyl acetate?"
No, we haven't been wetting the grids first. But that sounds like a reasonable thing to do. If Mollenhauer suggested it, it must be good.
*********************
"I also have one caution about using acetic acid to clean grids. If you leave the grids in the acetic acid too long, you'll get flakes of copper on your sections."
Yes, we're careful about that. One thing that may be contributing to the problem is that we're using the thin bar "Super 200" grids because we need to maximize the open space. These grids are difficult to wash as thoroughly as we'd like because there is less metal for the sections to adhere to and they are easily washed off the grid. But we've been using these grids and stain procedure successfully for years. We are looking for things in the laboratory environment that have recently changed. Our pay scale is not high on the list of suspects.
Greetings all! I am trying to use freeze substitution to retain soluble oligosaccharides in plant tissues, and am not yet convinced that I have been successful, partly because of embedding/infiltration problems (I think!). We have tried a variation of the tannic acid fixation with Kaeser's substitution cocktail, and stuff seems to look ok (tough to really assess when the scope has been down, unfortunately), although we have been advised that probably LR White is easier to work with than Lowycryl K4M, which is what we used the last time, and it doesn't section happily. Would like to try another run (just got some fresh LR White) and thought it would be useful to ask for some input from the group. It's probably of interest to more than me, but if folks prefer to respond directly, I will summarize the responses and post them. I'm certainly looking forward to your input!
TIA shea
S.Shea Miller Agriculture and Agri-Food Canada Centre for Food and Animal Research Central Experimental Farm Ottawa, Ontario Canada K1A 0C6 Phone: 613-759-1760 Fax: 613-759-1765 email: millers-at-em.agr.ca
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At 08:55 AM 2/2/96 -0800, you wrote:
} Could the UA somehow have been cross-contaminated with Pb? } Sometimes a weak wash with 0.02 N NaOH will help with Pb deposits-if that } is the source of the problem. } } } Doug Davis } ********************** Doug,
We've been pretty careful not to cross contaminate. We may have to resort to after-the-fact cleaning, but I would like to eleminate the problem before it occurs. Thanks for the suggestion.
I think there is something about the circumstances described over the last few days by Tina Carvalho that recalls some of the recent discussion of ethical values.
No matter how high and noble our calling, we all work within rules. Some of them are institutional, some professional and some come from the laws of our state and our nation. Whatever their source, however, the rules are what the rules are.
If we don't like the rules, we can work to change them, but none of us has a license to violate them. Auditors, internal or external, are in the business of finding violations of the rules, and some of them do a very good job of doing just that. All of us are tempted from time to time to bend or to break the rules under which we operate, sometimes for a very good purpose. In the case of our microscopy supplies business, for example, we may be asked to sell the components of a sputter coating system separately so that they can be bought out of a budget intended for consumables, when the rules say that the system is capital equipment, to come out of another budget. Is this right? Or is it a conspiracy to defraud? Does the noble purpose (the fact that the funds will go back to the sponsor) make any difference? How about the request that we predate or postdate the invoice, to get it into the "right" year?
It seems to me that we are all better off if we just force ourselves to live within the rules instead of trying to rationalize our efforts to get around them.
BTW, am I the only person who wonders which is the chicken and which is the egg when Tina comments about the lack of commercial EM facilities within her state?
Andy
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. 63 Unquowa Road Fairfield, CT 06430-5015 Ph: 1 203 254 0000 FAX: 1 203 254 2262 e-mail: AWBlackwoo-at-aol.com WWW--http://mail.cccbi.chester.pa.us/spi/spihome.html
Does anyone know where I might be able to purchase some used double tilt top entry specimen holders for a JEOL 4000 EX? Is there someone decommissioning a JEOL 4000EX that would like to sell their specimen holders?
Roseann Csencsits Argonne National Laboratory Argonne, IL 708-252-4977
Does anyone have a protocol for using (HPMA?) instead of propylene oxide and embedding coverslips directly in cell wells, and then using liquid nitrogen to remove the coverslip after plastic is polymerized?
Have you considered fixing the tissue in aldehyde and embedding in one of the other (hydrophobic) Lowicryl resins (e.g. HM 20, HM 23). They polymerize at lower temperatures and cut very well. We have had some success with soluble proteins in the pancreas and there are published examples of this method.
Here are a few references which might be of interest: van Genderen et al, 1991 J. Cell Biol. 115:1009-1019. Oprins et al, 1994 J. Histochem. Cytochem. 42:497-503.
There is also a recent paper in Microscopy Research & Technique which covers this subject but omits to refer to the above papers. The paper is in MRT, 1996 33:251-261. Look too at the work by Humbel & Schwarz which is cited in the MRT paper.
} 2 February 1996 } } From: Andy Blackwood } } To: Microscopy BB } } I think there is something about the circumstances described over the last } few days by Tina Carvalho that recalls some of the recent discussion of } ethical values. } } No matter how high and noble our calling, we all work within rules. Some of } them are institutional, some professional and some come from the laws of our } state and our nation. Whatever their source, however, the rules are what the } rules are. } } If we don't like the rules, we can work to change them, but none of us has a } license to violate them.
On the contrary. The idea that the existence of arbitrary rules defines ethical behavior is incorrect by construction. The classic examples, of course are in the realm of human rights violations -- where the ethical actions are explicitly those which are against the rules imposed by an unethical or irrational government.
It is *not* necesarily unethical to break rules, particularly when those rules are arbitrary, capricious, or in themselves unethical.
} It seems to me that we are all better off if we just force ourselves to live } within the rules instead of trying to rationalize our efforts to get around } them. }
When you call efforts to get around irresponsible regulations "rationalization" you beg the question that sometimes getting around the regulation is the only ethical thing to do. Sometimes finding an excuse for observing the regulations, and thus avoiding actions which involve personal risk, is the rationalization.
Ethics and rules are not orthogonal, but I, at least, refuse to let arbitrary bureacrats and supercilious functionaries get away with calling me "unethical" if I am not sufficiently obsequious to their dictates.
} Considering the number of comments and questions I received concerning my } recent recommendation of Q&A database software... } Don Grimes, Microscopy Today
I have used Q&A, and must agree with Don Grimes. Most database programs have a lot of unnecessary bells and whistles.
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
On February 20 was written:
} Subject: Pt TEM grids } } Does anyone make Pt TEM grids? They are needed for high temperature } experiments-other grids oxidize. } } Roseann Csencsits } Argonne National Laboratory } Argonne, IL } 708-252-4977 } } Pt grids are not in our printed catalog but are to be found in our all- electronic catalog on our WWW website as indicated below, in several different mesh sizes.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
I am in the proces of purchasing a digital image acquisition system for a Hitachi H7000 stem. To do this I need a digital beam control box to interface the microscope to the acquisition system. The later models, eg. Hitachi H7100 have this circuitry built in but the older models (H7000) need the DBC box which Hitachi made but no longer does. Does anyone out there have one that they don't use anymore or know of any extras collecting dust on a shelf somewhere? I am slightly desparate and have some money (not much) to pay for such a beast. Thanks in advance for info or sources, Hank Adams EML, New Mexico State Univ email: hadams-at-nmsu.edu phone: 505/6463600
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. wrote: "......It seems to me that we are all better off if we just force ourselves to live within the rules instead of trying to rationalize our efforts to get around them........".
Andy is correct. The rules and laws must be followed. Those we don't agree with we can work to change. But am I the only one who reads Andy's entire commentary (that only mentions what academics do to get around some regulation in order to purchase an item) and feels resentment? I have been approached many times by vendors who ask me to write lock-out specifications so I will buy from only one source. Other vendors have informed me that if I bought a certain dollar amount of supplies, they would throw in the instrument for free that uses those supplies. More than once I have successfully appealed to the State of Texas on a bid where an unscrupulous vendor misrepresents their instrumentation and/or the price. Some vendors are as eager to earn the academic dollar as the academic is to stretch what little he/she has. What about vendors/business people who appeal to the government and/or granting agencies to restrict the activities of the academics, thus reducing competition? Chuck Garber will readily inform you of the meeting that promulagated the rule that says instrumentation purchased with federal dollars cannot be used for commercial purposes. When that rule was made, there was no academic representation (the perspective of scientists at NSF/NIH is different than that of scientists in colleges and universities). Is that fair? What about the politically incorrect white vendor(s) who set up, or use, minority/women fronts in order to sell their products? Is there any noble purpose for vendors who abuse the system for profit and gain? A more important question would ask why is there such a strident militancy against academics among certain individuals at SPI? Let it out, guys, so maybe we can feel your pain......
Chuck
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
In response to the inquiry from anonymous subscriber, ratyrg-at-esvax.dnet.dupont.com, posted on 96-01-24 (regarding availability of cryomicrotomy courses), please be advised that AMC Group offers a series of intensive hands-on workshops on cryomicrotomy on a regular basis. We also offer other advanced workshops including the FIB cross-sectioning and wedge-polishing for site-specific TEM specimen preparation. For the workshops schedule and other informative material on the general subject of specimen preparation, please see our page on WWW at the Microscopy-Online site (http:// www.Microscopy-Online.com/). For further information, please contact me directly. Thanks.
} What about vendors/business people who appeal to the government } and/or granting agencies to restrict the activities of the academics, thus } reducing competition? } Charles J. Butterick (Chuck)
Or, as an actual experience, the SEM vendor who threatened--the correct word--to file a complaint with US Customs, and force a *previous* employer to pay a multi-thousand-dollar penalty if we didn't buy a US-made SEM. (Needless to say, we didn't, and they didn't, since it was a fatuous threat. And a permanent loss of business.) Phil Oshel (Not at U. Ill. at above the time)
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
Group: I am sure that many of you are familiar with the nifty microscopy images produced by David Scharf. If not, allow me to note that they have been featured in Life, Time, National Geographic, Scientific American, Discover, Natural History, Harpers, The Saturday Evening Post, etc. As it happens, I am in the process of producing a series of 16 of his best images - typically as 23 in x 19 in, HIGH quality, lithographic prints. Descriptions, etc. of the series is contained in the next issue of Microscopy Today, which will be mailed in a week's time. Should you, including our international associates, not be receiving the newsletter and have a potiential interest in the prints, kindly advise by eMail and I would be delighted to send you a no cost description of the series. And, should the publication of microscopy prints be an "enjoyable" hobby, I may extend the series. Should you have microscopy images that you consider worthy of reproduction, I would appreciate hear from you. Understanding that there is a bit of "commercialism" in this note, I hope that the uniqueness of the subject causes not many of you to be offended. Best regards to all, Don Grimes, Microscopy Today
Do you know anyone who might be interested in an ISI Super III-A SEM that's in excellent operating condition for only $8,000? We are in the St. Paul, MN area and would be happy to demo the scope in our location on your specimen. This workhorse would be an excellent instrument for a small college/lab/hobbyist or manufacturer. Contact me for more specs or info. Ask me about a $300 travel rebate offer.
} In the past I have used chloroform routinely for expanding thin sections. } Because of the hazardous nature of this chemical I tried a heat pen } (amps - .125), but it was totally ineffective. Would a more powerful } heat pen be effective in reducing compression ? I would appreciate } some feed back on this. } Thanks } Leo
The heat pen I use is fine as long as it is held close to the section and used for around 30 secs (longer for some sections).
Diana van Driel Dept Ophthalmology Sydney University 2006 NSW, AUSTRALIA
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Sixth Asia-Pacific Conference On Electron Microscopy.
Under the Auspices of The International Federation of Societies for Electron Microscopy (IFSEM) and The Committee of Asia-Pacific Societies for Electron Microscopy (CAPSEM).
The Sixth Asia-Pacific Conference on electron Microscope will be held at the Chinese University of Hong Kong, to be held on July 1-5, 1996.
Final call for abstracts.
The deadline for abstracts has been extend to 15th February, 1996. If you are close to the deadline, please use an express mail service to submit your abstracts. Faxed abstracts cannot be accepted.
The conference will provide a forum for the dissemination of new information on the application of scanning and transmission electron microscopy, diffraction and microanalysis in life and materials sciences. The conference will include scientific sessions (plenary lectures, symposia oral presentations and posters). There will also be an exhibition of scientific instruments together with workshops and open laboratories.
******************************************************************* Please contact the above address only. Please do not send requests for information to this e-mail address.
Message-ID: {199602051315.IAA18418-at-IndyNet.indy.net} To: Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
At 02:41 PM 2/4/96 -0500, you wrote:
} Dear Joiner, } If you find a solution to your staining problems, could you please let me } know? I have used the same receipe for UA and LC for over 20 years without } problems. But recently, have been haunted by the symptoms you relayed, with } always using ddwater and trying NaOH washes, different grid suppliers... I am } ready to record the position of the planets when I get good staining. I } wonder if such can be related to low humidity and increased static } electricity???????????? I hope you are able to solve this curse and that you } will share such with me. Needless to say, I will let you know if I have any } luck. } } Best regards, } } Robyn Rufner, Ph.D. } Deborah Research Institute, 20 Pine Mill Rd., Browns Mills, NJ 08015 } 609-893-1016, fax: 609-893-2441, email: rrufner-at-aol.com } ******************** Robyn -
I will indeed share what I've gleened from this thread, but as I told Dwight Beebe, not until you all have solved my problem. No, actually I will pass it on. That's what this listserver is for. The static charge sometimes found on grids has been suggested to be causing precipitation of the stain. If I can find one of those antistatic guns that we used to de-charge our LP's with, I'll try it, being the student of reason and logic that I am and staunch supporter of the scientific meathod. Then at the next full moon I'm going to nail a dead cat to the north side of an oak tree....
Mime-Version: 1.0 Pogany Lajos {pogany-at-power.szfki.kfki.hu} Content-Type: text/plain; charset=US-ASCII Content-Transfer-Encoding: 7bit Content-Description: cc:Mail note part
I'm also looking in to CDR's for archiving. I think we will get the Pinnacle 5040, a 2x write 4x read, which runs about $1300 ($1000 internal). HP makes one for about the same cost, Yamaha and Sony have models for a bit more $.
Any comments from people already using any of these?
Is anyone able to assist me in finding a CD ROM writer for image archieving in the price range of about 1000 $? Would be possibile to send me addresses where are those available?
does anyone know if there is a newsgroup or a listserver discussing problems of measuring Ca2+ with fura? Perhaps a group dealing with fluorescence microscopy in general would help.
Thanks for any help
-- Ralf Steinmeyer (Steinmeyer-at-mbox.biophysik.uni-hannover.de) UNI Hannover Herrenhaeuser Str. 2 Inst. f. Biophysik 30419 Hannover
We use Kodak lens cleaner for routine cleaning, plastic dropper bottles are refilled from the more affordable quart containers. Chloroform is reserved for obstinate deposits, like when someone immerses a non-DPX lens into the DPX or Permount.
A couple of microscope technicians that have worked on our equipment use lighter fluid. It dries without leaving a film and yet is claimed not to attack lens adhesives or coatings. Has anyone else used lighter fluid on lenses? Personally, I've only used to take scuff marks of vinyl flooring.
Regards,
Glen MacDonald Research Scientist Hearing Research Laboratories of the Virginia Merrill Bloedel Hearing Research Center Box 35-7923 University of Washington Seattle, WA 98195-7923 (206) 616-4156 glenmac-at-u.washington.edu
} Recent messages by Blackwood and Oliver are examples of two opposing points } of view about ethics and philosophy. On one side is the normative view: } that there exist absolute standards ('norms') for conduct. On the other } side is the relative view: standards depend on the situation.
I'm not so sure that it's a question of absolute versus situational ethics as much as *who* defines the "right" and "wrong." My position is not that ethics are situational; my position is that bureaucratic rules and ethics are not the same. Thus, noncompliance is not prima facie evidence that someone has done something "wrong." My primary concern is with the increasing criminalization of noncompliance, leading to allegations of "fraud," "scientific misconduct," and such.
I am not a scofflaw, and I certainly try to live within rules within reason. However, I don't mistake regulatory pronouncements for divine dispensation. Not confusing the two does not equate to the support of anarchy, nor does necessarily make it easy to rationalize any actions, as a different respondent suggests. Indeed, I might suggest the opposite -- that recognizing and accepting individual responsibility for individual ethical decisions independent of an external rulemaking bureaucracy might lead to more careful consideration of action, not less. Relying on legalistic justifications is perhaps not the last, but certainly one of the favorite refuges of a scoundrel.
} Let me remind the reader that we are not talking about human rights } violations, tyranny or murder here. We are only talking about money. It } really isn't worth doing more than 'blowing off steam'. I say follow the } rules that exist and work to change the rules that appear improper.
You bring up another interesting idea, that we are not talking about human rights, but, instead, "We are only talking about money."
In fact, we *are* talking about human rights. It isn't "only money" if a governmental agency, by its regulatory or enforcement rights, deprives you of your livelihood, appropriates your possessions, and criminalizes violation. When investigations result in allegations of fraud there is resultant criminal liability; the only question then is whether or not the regulatory agency wants to take you down bad enough to expend the effort. It may be only money, but when legal fees and seizures destroy life savings it is more than a mere inconvenience. When allegations of fraud or "scientific misconduct" based on technical violations of regulatory pronouncements leads to the destruction of a career, it is a nontrivial life event. For me, at least, it would not be "only money" at risk. As ex-Secretary Donovan said after the trials that destroyed his career in spite of finding him innocent, "Where do I go to get my reputation back?"
When the time and effort it takes to attempt to satisfy the voracious appetitite of a regulatory agency detracts from scientific investigation, patient care, or stifles innovation and experimentation, it is a loss to more than a single investigator. When regulatory demands and limitations force new intellectual exploration and development off the shores of the US, it hurts this nation badly.
In spite of all of this, I would take a more sanguine attitude about today's regulatory atmosphere were it not for the problem that, in my personal opinion, in many areas the regulatory morass is so byzantine and self-contratictory that it is fundamentally impossible to remain in compliance given limited time and resources to devote to doing so. In many a regulatory setting, particularly an investigation, the person being investigated is by no means considered "innocent until proven guilty." Instead, the assumption is that the worst possible interpretation is the correct one, and it is the burden of the person being investigated to prove his or her innocence. And the investigators have all the resources. It is no surprise that even the innocent plea bargain when they face financial ruin in the face of defending themselves against the charges of "fraud," "conspiracy," etc., etc., etc.
In such an atmosphere, simply trying to do one's best is just not good enough. A person who thinks he or she is working within the rules simply doesn't know all the rules. There are no innocent, only the untargeted, and obscurity is a weak defense.
I believe that I am one of the vendors that Chuck Butterick is referring to that offered *free* equipment with the purchase of a certain amount of supplies. It has been a a year or two since I offered the program to Chuck.
He is correct that the idea was to get around budget rules that allow unlimited purchase of supplies but no capital or equipment purchases. In this particular instance, if you bought the equipment and the supplies(film, in this case), paying the lowest available pricing, you would spend the same $ as you would if we gave you the hardware and you paid the premium price for the requisite cases of film.
I will admit that we have a vested interest in placing our equipment with customers so that we sell film. However, I would suggest that if our customers have a need for any of our products and due to various constraints are unable to acquire it through equipment purchase funding, we are working to meet the customers needs-we cant make the customer invest in equipment or participate in a program if they do not want to.
Yes, I am in sales and my livelihood depends on what I sell. I believe that what I sell offers my customers a better way of doing their job;more effectively and more efficiently-if I didnt, I'd quit.
Certainly I would agree that some vendors may have unscrupulous practices and I do not condone that. I am asking you to consider that the intent behind our program and possibly others is to benefit you-so you can have the tools you need to do your job.
John D. Warren Eastern US Sales Manager Helios Scientific Group Polaroid Corporation ______________________________ Reply Separator _________________________________
Andrew W. Blackwood, Ph.D. Structure Probe, Inc. wrote: "......It seems to me that we are all better off if we just force ourselves to live within the rules instead of trying to rationalize our efforts to get around them........".
Andy is correct. The rules and laws must be followed. Those we don't agree with we can work to change. But am I the only one who reads Andy's entire commentary (that only mentions what academics do to get around some regulation in order to purchase an item) and feels resentment? I have been approached many times by vendors who ask me to write lock-out specifications so I will buy from only one source. Other vendors have informed me that if I bought a certain dollar amount of supplies, they would throw in the instrument for free that uses those supplies. More than once I have successfully appealed to the State of Texas on a bid where an unscrupulous vendor misrepresents their instrumentation and/or the price. Some vendors are as eager to earn the academic dollar as the academic is to stretch what little he/she has. What about vendors/business people who appeal to the government and/or granting agencies to restrict the activities of the academics, thus reducing competition? Chuck Garber will readily inform you of the meeting that promulagated the rule that says instrumentation purchased with federal dollars cannot be used for commercial purposes. When that rule was made, there was no academic representation (the perspective of scientists at NSF/NIH is different than that of scientists in colleges and universities). Is that fair? What about the politically incorrect white vendor(s) who set up, or use, minority/women fronts in order to sell their products? Is there any noble purpose for vendors who abuse the system for profit and gain? A more important question would ask why is there such a strident militancy against academics among certain individuals at SPI? Let it out, guys, so maybe we can feel your pain......
Chuck
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
This is a request for information from colleagues experienced with the thinning of TEM specimens held by a Gatan Disc Grinder against South Bay Technology (or 3M Imperial) diamond abrasive films. First some information:
My TEM specimens are taken from joints that have been diffusion bonded by Hot Isostatic Pressing, using the method described in J. Mater. Sci. 29 (1994) 4404-4414. The diffusion couples are as paired semicylinders fitted into a small metal tube (3 mm outer diameter) of stainless steel or nickel. This tube acts as an encapsulation for the isostatic argon pressure (200 MPa) during HIP, and afterwards the capsule is simply sliced with diamond saw before thinning,leaving the capsule as a stabilizing collar around the diffusion couples avoiding cracking for materials having very different coefficients of thermal expansion, such as silicon nitride and Ni-base superalloys. The materials joined so far include Ni-base superalloys (both single crystal and polycrystalline), silicon nitride, titanium nitride, as well as CMC (Si3N4/TiN) and MMC (TiN/Ni) compositionally graded materials (FGM´s).
In June 1995, I participated in a very informative TEM Spec. Prep. Course (lead by Ron Anderson, IBM, East Fishkill Lab, NY, USA) held at BAL-TEC AG in Liechtenstein. After that, I decided to change the type of thinning abrasives from diamond spray on a soft cloth to SBT diamond abrasive films with fixed diamonds (30µm, 15µm, 6µm, 3µm, 1µm and 0.5µm grain sizes), finally followed by colloidal silica on a soft cloth.
However, due to the usefulness of retaining the stabilizing collar of capsule tube, I still want to use the Gatan Disk Grinder (GDG) for parallel thinning, instead of the otherwise impressive SBT Tri-pod polisher demonstrated at the course. Regarding the glue for fixing the specimen on the specimen mount of the GDG, I have changed from wax to Crystalbond 509, in order to avoid remaining films after cleaning in aceton. Finally, I intend to perform low-angle ion beam milling using the BAL-TEC RES 010, possibly preceeded by dimple grinding.
Based on this, I would like your comments on the following questions:
-What are the experiences from the combination Gatan Disc Grinder and South Bay Technology (or 3M Imperial) diamond abrasive films (8´´, plain back), adhering by capillary forces to a slowly (50-100rpm) rotating glass plate?
-Is it always (or exclusively for ductile materials) necessary to use all of the above grain size steps and/or remove a damage layer from the previous step corresponding up to three times the grinding grain size? (If so, given a cutting wheel such as Struers 330CA having 68µm (Grit #220) diamonds, it seems necessary to cut samples thicker than 400µm!)
-Is it necessary/unsuitable to press the GDG against the diamond abrasive film, or is the weight of the GDG itself (660g) sufficient? (If the specimen area would carry the total weight, it would correspond to a pressure of approx. 1MPa (135psi).
-Is it beneficial to rotate the GDG itself during polishing?
-Is it beneficial to grind down the specimen continously or incrementally (50µm steps are recommended by Gatan). If stepwise, should the steps be related to the diamond film grain size?
-Is the fit between the specimen mount and the GDG itself crucial for the integrity of the glued specimen? Which tolerances (after wear) can be tolerated?
Thanks in advance,
/Richard ________________________________________________________________________ Dr. Richard Larker, Ph.D. Phone: +46 920 91107 Research fellow Fax: +46 920 99309 Division of Engineering Materials Lulea University of Technology E-mail: Richard.Larker-at-mb.luth.se S-97187 Lulea, SWEDEN ________________________________________________________________________
I am going to be teaching a mixed TEM/SEM course to undergraduates in Materials Science next quarter. I am sending this note out to ask for comments/suggestions about textbooks, both from people who have taught and those who have been at the recieving end.
NOTE: For obvious reasons, please respond directly to me rather than everyone on the listserver unless you really intend to do the latter.
Message-Id: {9602060920.AA08107-at-smtp-mail.well.ox.ac.uk} X-Sender: levy-at-well.ox.ac.uk X-Mailer: Windows Eudora Version 1.4.4
Please would someone send me the list serv address for subscribing to this list.
Thank you
Elaine Levy -------------------------------------------------------------- Elaine Levy PhD Cytogenetics Laboratory Wellcome Trust Centre For Human Genetics Windmill Rd., Oxford OX3 7BN
tel 01865 740022 fax 01865 742186 email elaine.levy-at-well.ox.ac.uk -----------------------------------------------------------------
our group has received 2 bottles of CIT ALCATEL 220 diffusion pumps oil for free. We have no information at all about how good it would be for our JEOL SEM and we have found no cataloges around. If any of you knows something about vapor presion of this oil, or has experience using it, and has a little time for writing to us, we would appreciate it very much. Thanks in advance,
Silvia Montoro csedax-at-arcride.edu.ar
Regional Center for Research and Development Santa Fe Argentina
} ... recognizing and accepting individual responsibility } for individual ethical decisions ...
To me this seems to be the key; together with a well estabished line of accountability (very important). We will still make mistakes, but hopefully won't be alone in them...
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
I would like to set the record straight about "militancy against academics among certain individuals at SPI" and also, the record with regard to the true complexion of those at NSF who were resonsibile for the drafting and implimentation of NSF Important Notice 91.
First, there is no such "militancy", indeed our firm relies on the graduates of the nation's academic institutions just like any other employer. Our future business prospects in fact are closely intertwined with the future prospects of our academic customers. So any suggestion to the contrary, as they say on the other side of the ocean, is just pure rubbish.
If there is indeed any "militancy" at all, it is on the part of a small handfull of individuals and departments who are literally running businesses out of their university laboratories and who feel threatened should anyone suggest that what they are doing is not right.
So surely there is concern about anyone using university equipment in competition against private sector laboratories such as our own. For the most part, people I know in academia are on "our" side of this issue and hold the view that the university's instrumentation in fact should not be used this way. They seem to feel that those projects that are basic and fundamental in nature and suitable for inclusion in a student's thesis (e.g. those projects that do contribute to educational objectives) take up the available time for their instrumentation anyhow and that outside commercial work just gets in the way and interferes with the progress of the students and the quality of the work being done. So those who advocate some contrary view are clearly on the wrong side of the tracks on this issue, and are also on the wrong side with respect to their own academic peers.
Second, at the time of the drafting of NSF Important Notice 91, the Director of NSF was Prof. George Pimentel, perhaps one of the most respected and published academicians ever to head NSF. He came to NSF from the Chemistry Department at the University of California-Berkeley. He was an "academic" and represented "academic interests" in every sense of the word. To suggest otherwise is to misrepresent Dr. Pimentel's attitudes, perspectives, and outlook.
The Associate Direrctor at the time was Dr. Donald Langenberg. Prior to his position at NSF he was a full professor of physics (at the U. of Pennsylvania, if I remember correctly), and after his tenure at NSF moved on to serve as President of the American Association for the Advancement of Science, the Chancellor (I think that was the postion) of the University of Illinois - Chicago and from there to his present position of Chancellor of the University of Maryland system, with over sight over all state universities in the State of Maryland.
Both Drs. Pimentel and Langenberg were in attendence at most of the meetings that led up to the drafting of NSF Important Notice 91. And their participation in the drafting of the document was certainly in no way to be thought of as being passive. So for anyone to suggest that in some way the academic community was not properly represented is again "pure rubbish".
But intentional or not, it is obvious that there are still some who would try to discredit the NSF Important Notice 91 document in every possible way, in this case, by suggesting there was a lack of academic representation among those involved with its drafting and implimentation.
With regard to some of Chuck Butterick's other comments, unethical behavior of any type should not be tolerated in any environment where high quality research is being conducted. Vendors guilty of the kinds of infractions described in Chuck's posting should be reported to the appropriate procurement officials who have in place mechanisms by which such abuses of process can be rooted-out and stopped. Of course it does take two to tango and therefore the other side of the equation, that is, the customer who is encouraging such inappropriate practices should also be reported. While the wheel of "justice" might move slowly, the wheel does move, but please don't paint the behavior of the entire vendor community with the same brush. The electron optics industry of manufacturers and distributors by and large is highly professional and honest and when instances do occur that do not fit this pattern, there is a process by which one can keep that kind of circumstance from recurring in the future.
As a final comment, for anyone not personally familiar with NSF Important Notice 91, I would be happy to FAX you a copy, just send me your FAX number.
Chuck (Garber) ====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
At 08:21 AM 2/6/96 EST, you wrote: } Joiner- } If you *do* find an old Zerostat gun (which is a little like searching for } brontosaurus chow), it's probably not going to be any good any more. The same } effect can be acheived by using glow discharge. Have you got that on an old } evaporator or coater? Good luck! } Steven } } ************** Yes, I do; although the evaporator ain't no spring chicken.
Message-Id: {199602061557.JAA23186-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 09:06 AM 2/6/96 -0500, you wrote: } Good Morning. } } Further to the note I sent you last week where I fingered the grids as my } most likely suspect for the introduction of Pb hexagons: } } My understanding is that these things are made by a plating method. We had a } program here for a while looking at the microstructure of thin film magnetic } media for one of our then subsidiaries. The magnetic media is deposited on } plated substrates. } } One of the things we struggled with, is that the plating procedure was done } in baths contained by tanks soldered together with (you guessed it) Pb. The } Pb impurities found their way into the plated metal (concentrating at the } surface), and when I would microtome these things, I'd get hexagons aplenty! } } Perhaps the source of contamination in the grids? }
********************* That has been proposed (the grids being the source) and we are gearing up to improve our grid washing.
On June 22, 1995, Chuck Garber wrote, "A special committee, made up of top NSF officials (the late Dr. George Pimentel played a key role in the formulation of the final document as did also Dr. Donald Langenberg, now Chancellor of the University of Maryland system), lawyers representing the NSF (Mr. Charles Herz who is still at NSF in that same capacity), members of the independent laboratory community (including myself) plus some other knowledgeable people came up with the first draft of NSF Important Notice 91."
I still stand by my statement, no academic representation existed. Chuck Garber was once a part of academics, but he doesn't represent academic interests. NSF scientists, though academics at one time, are representing NSF and no one else. That committee appears to have been a stacked deck of cards. This is a democracy. Bad law can be overturned.
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
Low temrerature embedding/infiltration is not an easy procedure. The concetration and the distribution of the soluble proteins play an important role of successful LT embedding. I have had some success to retain soluble proteins in different type of plant tissues. Enzyme activity in vacuoles and in cell wall,using post-embedding techniques is our interest. Lowicryl resins: HM23 and K4M, as well as LR White were used at different temperatures and times (-80 to -20C; 2days to 2 weeks) after "mild" GA fixation OR cryo- fixed plant tissues, using PLT techniques (dehydration at progressively lower temperatures). LR White, after UV polymerization developed very small air (?) "packets". It doesn't section happily, yet not a major problem. "An Osmium-free Method of Epon Embedment That Preserves both Ultrastructure and Antigenicity for Post-embedding Immunocytochemistry", Phen, K.D., Rustioni.A., and Weinberg, R.J., J. Histochem.Cytochem. 42, 1995. You may adopt this procedure to LR White. It should work well. Good luck! Any other ideas, please let me know. Regards,
Laszlo J. Veto Electron Microscopist Agriculture and Agri-Food Canada Summerland, BC Ph: 604-494-7711 Fax: 604-494-0755 e-Mail: veto-at-bcrssu.agr.ca
} Robyn - } } I will indeed share what I've gleened from this thread, but as I told Dwight } Beebe, not until you all have solved my problem. No, actually I will pass it } on. That's what this listserver is for. The static charge sometimes found on } grids has been suggested to be causing precipitation of the stain. If I can } find one of those antistatic guns that we used to de-charge our LP's with, } I'll try it, being the student of reason and logic that I am and staunch } supporter of the scientific meathod. Then at the next full moon I'm going to } nail a dead cat to the north side of an oak tree.... } } * * Joiner Cartwright, Jr. * *
Barring a Zero-Stat gun, which I've still seen advertised somewhere, try touching your charged grid to a bit a metal on a grounded piece of electrical equipment. It's safe (this is how some powerbars drain your personal static charge so you don't zap your computer). Phil
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
I received a used ISI DS-130 SEM equipped with a cathodoluminescence (CL) detector. This detector has no ellipsoidal mirror coupling to it. I guess this must be a "diode" detector which can detect CL directly from the sample.
(1) Does anyone have information about this type of detector ? How does it compare to the detector with an ellipsoidal mirror ?
(2) Does anyone have experience taking CL images of quartz? I know we cannot focus the CL, but at least we can use proper settings (by adjusting accelerating voltage, beam current, etc) to optimize the CL resolution. Thanks.
Long Liang ARCO EPMA/SEM Lab lliang-at-is.arco.com
} On June 22, 1995, Chuck Garber wrote, "A special committee, made up } of top NSF officials (the late Dr. George Pimentel played a key role in the } formulation of the final document as did also Dr. Donald Langenberg, now } Chancellor of the University of Maryland system), lawyers representing the } NSF (Mr. Charles Herz who is still at NSF in that same capacity), members } of the independent laboratory community (including myself) plus some other } knowledgeable people came up with the first draft of NSF Important Notice } 91." } } I still stand by my statement, no academic representation existed. } Chuck Garber was once a part of academics, but he doesn't represent } academic interests. NSF scientists, though academics at one time, are } representing NSF and no one else. That committee appears to have been a } stacked deck of cards. This is a democracy. Bad law can be overturned. } } } } } Charles J. Butterick (Chuck) } Electron Microscopy Center } Department of Cell Biology } and Biochemistry } Texas Tech University Health } Sciences Center } 3601 4th Street } Lubbock, Texas 79430 } } vox (806) 743-1633 } fax (806) 743-1219 } email emccjb-at-ttuhsc.edu or } chuck-at-micron1.lubb.ttuhsc.edu
Dear Chuck's (Butterick and Garber),
I really believe that this sort of dialog (?!) does not belong here.
Thank you for the excitement, though!!!
Best regards
Massimo Sassaroli, D.Sc. Dept. of Physiology & Biophysics Box 1218 Mount Sinai School of Medicine 1 Gustave L. Levy Pl. New York, NY 10029-6574
We would like to know if anyone has an opinion on the effectiveness of using a silver screen and double projectors for 3D imaging. We do not want to commit ourselves to buying a screen (which is quite expensive) and then find it has been a waste of money.
We intend to use the system for illustrating talks to members of the the public visiting the Unit and also for student classes. We would be projecting mainly SEM images.
We would like to hear from anyone who has tried this or any other alternative method of displaying 3D images to groups of people.
Thanks in advance,
Mark Gould
Richard Easingwood South Campus Electron Microscope Unit Otago Medical School PO Box 913 Dunedin NEW ZEALAND
I am seeking facilities for SEM-EDS and/or SEM-WDS in proximity to Berkshire county, Massachusetts (to include western MA, southern VT and eastern NY). Anyone knowing of a commercial, industrial or academic facility in this area, please contact me directly.
Dear Dr. Hendriks, (and other colleagues interested in the topic related to the first question (Q1) below),
I have the following comments/questions (C1-6) caused by your answers (A1-6) to my questions (Q1-6):
Q1: What are the experiences from the combination Gatan Disc Grinder and SouthBay Technology (or 3M Imperial) diamond abrasive films (8´´, plain back), adhering by capillary forces to a slowly (50-100rpm) rotating glass plate?
A1: While I have not used the Gatan tool myself, I can answer you in terms of using our SBT Polishing Fixtures. Other than the Tripod Polisher (R) we also have other fixtures which are similar in size and weight to the Gatan. We have had very good experience in having the films remain firmly attached - which I presume is your question. We typically recommend that people use the plain back films as they adhere well, are easy to remove without damaging them and then can be re-used. This is important as the films are not inexpensive! You may also want to note that the bottom of the Gatan fixture is stainless steel (I believe) and it will wear when polishing with diamond. The SBT fixtures described above have tungsten carbide feet at the bottom to help resist wear. However, even the tungsten carbide will wear when polishing diamond. For final polishing on samples less than a few microns thick, it is best to use a much slower wheel - ie less than 5 rpm.
Comment 1: In my tests so far, the SBT diamond films were firmly attached to the wet glass plate by squeezing the water out simply by pressing a PMMA ruler in several directions on the films. In spite of this, the specimen (glued to the mount by Crystalbond 509 at 130 C) sticked several times to the 30 micron diamond film and loosened from the mount, causing expensive scratches in the film! I tried to reduce both the incremental steps (from initially 25 micron down to 5 micron steps) and the rotational speed from 100 rpm to 50 rpm, but with limited success. I experienced the same problem with the 6 micron diamond film (but not with the intermediate 15 micron!) when using 2.5 micron steps. The feeling during this "seizure" was that the whole GDG was sucked (by capillary forces?) firmly to the film, twisting it loose, causing scratches and loosening of the specimen from the mount. This kind of experiences did not occur earlier when using diamond spray on soft cloths! Perhaps I have used a little too high pressure on the GDG, but the unpleasant tendency for it to "follow" the film was noticeable even without any extra load. It was even noticeable when trying without any specimen mount present in the middle of the GDG! This was peculiar, since the instructions inside the cover of the Gatan Disc Grinder 623 specify the following: "Polish specimen preferably using 3M Imperial Lapping films". Furthermore, the instructions recommend as large increments as 50 micron! Concerning your comment that the WC-Co feet of the SBT fixtures should wear less than the large SS plate of the GDG does; supposing that is true, what does it imply for the wear of the soft Delrin feet of the Tripod? How is it then possible to know that the micrometer is not "underestimating" the thinning of the specimen? What is the influence of the specimen/feet area ratio?
Q2: Is it always (or exclusively for ductile materials) necessary to use all of the above grain size steps and/or remove a damage layer from the previous step corresponding up to three times the grinding grain size? (If so, given a cutting wheel such as Struers 330CA having 68 micron (Grit #220) diamonds, it seems necessary to cut samples thicker than 400 micron!)
A2: The short answer is NO. You do not need to use every grain size. You do however, need to be mindful of the 3x rule when polishing. I assume that you are interested in the microstructure of the sample being polished. You can polish a sample from 500 microns thick down to 5 microns thick by using only 0.5 micron film. The problem is that it will take you many, many, many hours and consume many pieces of film. Reason the many steps are used is to minimize the polishing time. You can obviously remove material much faster with a 30 micron film than with a 1 micron film, but you will also cause more damage. The polishing sequence is determined by 1) amount of material to be removed 2) desired final thickness 3) budget for film 4) level of patience. I do not fully understand your example of the Struers 330CA so I won't address that here.
C2: The example of the Struers 330CA diamond cutting wheel, which I have used for slicing my samples up to now, was after some investigation found to have a diamonds of 68 micron size (Grit #220), and if this causes 3x68 micron of damage on each side, and you follow the 3x rule with 30-15-6-3-1-0.5 micron diamond films, you simply need to cut thicker (} 740 micron!) than I have been doing so far (400 micron) to end up with any specimen thickness at all! However, may it be the case that the cutting wheel causes less than the anticipated 3x diamond grain size damage, due to the low level of normal force against the surface during cutting, when compared to polishing conditions? Regarding thinning of ceramics like silicon nitride, which diamond grain size steps might be omitted ? Can any step be omitted for Ni-base superalloys?
Q3: Is it necessary/unsuitable to press the GDG against the diamond abrasive film, or is the weight of the GDG itself (660g) sufficient? (If the specimen area would carry the total weight, it would correspond to a pressure of approx. 1MPa (135psi)).
A3: In general, it may be advisable to add some additional pressure during the rough grinding stages, however, the during final polishing stages it is best to minimize the load on the sample. Unfortunately, the Gatan Disc Grinder has no facility which will allow you to reduce the specimen load below the weight of the fixture. If you have had problems with samples cracking when thinning at the final stages, excessive load is probably the problem.
C3: Due to the experiences given in C1 above, I am not confident with adding any extra pressure at all! Regarding the thickness of the water film between the GDG and the diamond film, are there any estimations of the "regimes of lubrication" (hydrodynamic, partial or boundary lubrication, as related to the diamond grain size) below the GDG or SBT polishing fixtures, or even below the specimen itself?
Q4: Is it beneficial to rotate the disc grinder itself during polishing?
A4: Yes, for several reasons. It is best to continually introduce new cutting faces will increase removal efficiency and also reduce preferential scratching in the surface. However, in some instances, people will continue polishing in 1 orientation per grit size. This will provide a scratch pattern in one direction. When changing to the next size film, you can rotate the sample 90 degrees and see a new scratch pattern develop. Once the new scratch pattern covers the surface, you know that you have removed the scratches from the previous grit size - but not the damage! Remember the 3x rule.
C4: For the GDG, the specimen mount may not be unambigously prevented from rotating relative to the GDG orientation; is this also the case for the SBT polishing fixtures that you have described?
Q5: Is it beneficial to grind down the specimen continuously or incrementally?
A5: This is a question which is peculiar to the Gatan Disc Grinder. With the Gatan Disc Grinder your specimen is mounted to a post which DOES NOT freely float inside the larger outside ring. Polishing is accomplished by extending the sample below the surface of the larger outside ring. This means that the entire weight of you fixture is resting on your sample. There is a great likelihood that you will damage your sample and/or round the edges of your sample. Because of this inadequacy, Gatan recommends an incremental procedure which minimizes the effect, but maximizes the effort! In more thoughtful designs (such as the South Bay Technology Fixtures :-)) the sample is mounted to a free floating rod which is gravity fed into the abrasive surface. The load on the specimen is thus spread across the entire base of the fixture. Edge rounding is eliminated because the edges of the sample are protected by the base of the fixture. Using this type of fixture, the incremental polishing is not necessary.
C5: How are the SBT polishing fixtures designed to stop further thinning of the specimen after reaching the intended thickness in each step?
Q6: Is the fit between the specimen mount and the disc grinder crucial for the integrity of the glued specimen? What tolerances can be tolerated?
A6: Think of the disc grinder as a 2 part device 1) the piston with a sample mounted at the bottom 2) an outside ring with a center hole. The fit between the piston and the center hole should be very precise. This precision is increased by making a longer contact area between the 2. Unfortunately, the Gatan Disc Grinder has a very short contact area which will magnify any problems. As the Gatan piston/hole are nearly in contact with the abrasive surface, I would think the chance for abrasive wearing away the center hole is rather likely. The sample mount is replaceable so that part is not a big problem. I would think that if you do wear the center hole uniformly, you could make special sample mounts to correspond to the new center hole size. Certainly the accuracy in these 2 areas is crucial - especially when polishing very thin. I did notice that you are familiar with Tripod Polishing and that you said you did not want to do that because you wanted to retain the stabilizing collar. Since you attended Ron Anderson's course, he probably spoke to you primarily about "wedge polishing". You may like to know that it is also possible to use the Tripod Polisher (R) for parallel polishing by mounting the sample in the center of the 3 micrometers. In fact, this was the way Tripod Polishing originated. The "L" brackets and wedge polishing techniques were later developments. You also mentioned that you now use Crystalbond 509 for mounting your sample to the holder. As a matter of interest, we supply an identical product called QuickStick 135 which can be purchased in a package of 20 small (3" x .25" x .25") unwrapped sticks under part no. MWH135. An alternative may be to use "super glue" or any cyanoacrylate. It bonds quickly and removes completely in acetone. The only problem is that removing it is somewhat unpredictable. Sometimes it can come off quickly, other times it may take quite some time. It is also important to be sure to use fresh super glue. We use super glue for wedge polishing of our TEM samples. You can pick it up at a local hardware store. If you would like information on any of our products, you can contact me directly or you may contact our office in Sweden, Tudor Barnard (snip)
C6"+": How useful is the Tripod for parallel (not wedge) polishing, compared to the other SBT polishing fixtures that you have described? Regarding your representative in Sweden, Tudor Barnard, I bought my SBT diamond films through him, but he was at that time (august ´95) not able to find a cyanoacrylic glue that was soluble in acetone (but only in di-methylformamide or tetra-chlormethane, which I consider to be far more harmful substances!)
Thanks in advance,
/Richard ________________________________________________________________________ Dr. Richard Larker, Ph.D. Phone: +46 920 91107 Research fellow Fax: +46 920 99309 Division of Engineering Materials Lulea University of Technology E-mail: Richard.Larker-at-mb.luth.se S-97187 Lulea, SWEDEN ________________________________________________________________________
We have used the silver screen method of projecting 3-D slides of all varieties for over 10 years. We find it by far the best method for displaying 3-D material to a large group of people. We use separate projectors with cross polarizers (I have heard of but not seen large screen projector wich takes a single slide containing two images).The down side of the procedure is the time it takes to align the two projectors and the necessity that all slides be exactly aligned in the same way. Nothing will make an audience sea sick faster than realigning projectors between stereo-pairs to accomodate different alignments. We sometime spend an entire day making sure a group of slides is all set up similarly. For this reason, for showing one or two people our stereo work we use premounted pairs in small, hand held viewers. However, this is not nearly as dramatic as a huge image on a screen. Any one who has ever stood in front of an audience and presented a stereo presentation will also tell you that the second down side of the method is trying to keep your composure while looking out at an audience full of people wearing silly dark glasses in darkened room.
I hope this helps-
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
On Wed, 7 Feb 1996, Richard Easingwood wrote:
} } We would like to know if anyone has an opinion on the effectiveness of } using a silver screen and double projectors for 3D imaging. We do not want } to commit ourselves to buying a screen (which is quite expensive) and then } find it has been a waste of money. } } We intend to use the system for illustrating talks to members of the the } public visiting the Unit and also for student classes. We would be } projecting mainly SEM images. } } We would like to hear from anyone who has tried this or any other } alternative method of displaying 3D images to groups of people. } } Thanks in advance, } } Mark Gould } } } Richard Easingwood } South Campus Electron Microscope Unit } Otago Medical School } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } SOUTHERNMOST E.M UNIT IN THE WORLD } } } } } } } } } } } }
+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+ Simone T. Boesch {simone.t.boesch-at-uibk.ac.at} Dept. of Zoology, University of Innsbruck Technikerstrasse 25, A - 6020 Innsbruck, Austria phone: (43)-512-507-6171, fax: (43)-512-507-2930 +-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+-+
A simple way of projecting stereo images is to get a friendly photographer to double expose your stereo pair through red and green filters onto slide film. All you need then is to hand out the old red green glasses and appeal for them to be returned! Its cheaper that way!
The other main item that you need to know is that there is a convention as to which way around the exposures are made so as to register re. the red and green and whch way around the glasses are held.
Keith Ryan Plymouth Marine Lab. Citadel Hill Plymouth PL1 2PB, England
Does anybody know if Gatan has a WWW-site with FAQ´s on the net, or even just an E-mail address?
Thanks in advance,
/Richard ________________________________________________________________________ Dr. Richard Larker, Ph.D. Phone: +46 920 91107 Research fellow Fax: +46 920 99309 Division of Engineering Materials Lulea University of Technology E-mail: Richard.Larker-at-mb.luth.se S-97187 Lulea, SWEDEN ________________________________________________________________________
We are encountering problems in disposal of uranium salts as our site safety officer considers it as a mixed/radioactive waste. I wonder what is the experience of other microscopists regarding radioactivity/disposal of uranium compounds.
A student in Chemical Engineering wants to examine pore membranes in the SEM. Currently we are looking at the membrane surfaces but he was wondering about viewing the membranes in cross-section. Does anyone know a method to prepare such a section? Please respond to his email address at the University of Missouri at c517837-at-showme.missouri.edu or you can contact me directly.
Thanks in advance.
Lou Ross
101 Geological Sciences Bldg. University of Missouri Columbia, MO 65211 (314) 882-4777, 882=5458 fax
We have recived a JEOL 100B electron microscope (Series 155037-30) as a gift from University of Pennsylvania. Although this microscope is in good conditions, it needs complete rehabilitacion. We have highly qualified personnel to do this job but we found that the instruction manual and circuits diagrams we have recived, do not much with the specifications of the microscope. If somebody has these manuals from JEOL 100B electron microscope (with the series number higher than ours) which is not longer useful, we can do good use of them in this part of the third world.
Dra Patricia Pons Centro de Microscopia Electronica Universidad Nacional de Cordoba Cordoba - Argentina
Does anyone have information on TMAH as a back-etch for silicon devices? I have one paper "A New Back-Etch for Silicon Devices" P. Malberti, M. Ciappa , P. Scacco. Also, in this paper it refers to a "bain marie". Can anyone tell me exactly what it is? I am assuming it is like a double boiler.
Terri Mengelt Motorola COM 1 Phoenix, Arizona 85201
I would like to thank everyone who responded to my earlier email (see the end). A number of you were interested in the responses, which I am including here (with editorial comments omitted). As a follow up, I heard almost exclusively from faculty and only one student - how about a few more comments from the students (I will preseverse anonimity).
This is a list of the responses:
4 Responses suggesting (but SEM only) "Scanning Electron Microscopy and X-Ray Microanalysis", by Goldstein, et.al., Plenum Press
3 Responses suggesting "Electron microscopy and analysis" P.J.Goodhew and F.J.Humphries, Taylor and Francis 1988.
"Light and Electron Microscopy" by Slaytor and Slaytor.
"Scanning Electron Microscopy and X-Ray Microanalysis" by Robert E. Lee
"Scanning and Transmission Electron Microscopy," by Flegler, Heckman and Klomparens (W.H. Freeman and Co., 1993)
"Electron Beam Analysis of Materials" by Mike Loretto
---- Copy of Original Message -----
I am going to be teaching a mixed TEM/SEM course to undergraduates in Materials Science next quarter. I am sending this note out to ask for comments/suggestions about textbooks, both from people who have taught and those who have been at the recieving end.
NOTE: For obvious reasons, please respond directly to me rather than everyone on the listserver unless you really intend to do the latter.
We've got a minor glitch in the subscribe/unsubscribe system. Unfortunately, I'm on vacation and too far away to fix the link. This just means there will be a delay if your trying to unsubscribe. I'll be logging in remotely and doing manual updates, but the connections are sometimes slow from here.
I'll be back in town the middle of next week and we should be back to normal.
Sorry for the inconvenience
Nestor Your Friendly Neighborhood SysOp
Weather report: 34 C, partly cloudy, and the view on the beach is great. ;-)
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At 05:46 PM 2/6/96 -0500, you wrote:
} Dear Chuck's (Butterick and Garber), } } I really believe that this sort of dialog (?!) does not belong here. } } Thank you for the excitement, though!!! } } Best regards } } Massimo Sassaroli, D.Sc. } ********************************
Massimo -
At the risk of adding UNNECESSARY fuel to the fire, I'm going to disagree with you. These are issues that are indeed important, and if these two gentlemen (and they ARE gentlemen) can hash them out without resorting to nastiness, we will all benefit. It is easy for us academics to overlook the position of the commercial/for profit operator, and I assume that the converse is also true.
I would point out that there may be more than two sides to the question, or at least an intermediate position. My lab, for example, is in a medical school and I work WITH investigators who pay me for my services with their grant money. My lab receives no direct grant funding, either in its capital set-up and improvements or its operation. The only financial support we get is what we earn in fee-for-service.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
I am a microscopist with 21 years experience in Scanning Electron Microscopy in materials science. I am seeking a new position as a lab supervisor or a lab support technician. If you have or know of a position available that would utilize these basic qualifications, and to receive a copy of my resume, please contact me:
We've got a minor glitch in the subscribe/unsubscribe system. Unfortunately, I'm on vacation and too far away to fix the link. This just means there will be a delay if your trying to unsubscribe. I'll be logging in remotely and doing manual updates, but the connections are sometimes slow from here.
I'll be back in town the middle of next week and we should be back to normal.
Sorry for the inconvenience
Nestor Your Friendly Neighborhood SysOp
Weather report: 34 C, partly cloudy, and the view on the beach is great. ;-)
Please publish the following information in the MicroWorld e-mail newsletter:
1996 Lehigh Microscopy Short Courses June 10-21, 1996 SEM, X-ray Microanalysis, AFM and SPM, AEM Contact: Sharon L. Coe Department of Materials Science & Engineering Whitaker Labaoratory 5 East Packer Avenue Bethlehem, PA 18015 610/758-5133 (phone) 610/758-4244 (fax) slc6-at-lehigh.edu http://www.lehigh.edu/~inmatsci/Microscourses.html
Thanks you for your help.
Sharon L. Coe Department of Materials Science & Engineering 5 East Packer Avenue Lehigh University Bethlehem, PA 18015 610/758-5133 e-mail: slc6-at-lehigh.edu
I have projected stereo images successfully by creating red/green anaglyphs on transparency material. We took our digital images, combined them using PhotoShop or similar program, and printed them on a dye-sublimation printer. Using a standard overhead projector and the red/green glasses, the images greatly impressed my audience. The dual projector system seems a bit specialized.
Henk
} We would like to know if anyone has an opinion on the effectiveness of } using a silver screen and double projectors for 3D imaging. We do not want } to commit ourselves to buying a screen (which is quite expensive) and then } find it has been a waste of money. } } We intend to use the system for illustrating talks to members of the the } public visiting the Unit and also for student classes. We would be } projecting mainly SEM images. } } We would like to hear from anyone who has tried this or any other } alternative method of displaying 3D images to groups of people. } } Thanks in advance, } } Mark Gould } } } Richard Easingwood } South Campus Electron Microscope Unit } Otago Medical School } PO Box 913 } Dunedin } NEW ZEALAND } } Telephone: 64-03-479 7301 } Facsimile: 64-03-479 7254 } } SOUTHERNMOST E.M UNIT IN THE WORLD } } } } } } } } }
Henk Colijn colijn.1-at-osu.edu OSU Campus Electron Optics Facility ------------------------------------------------------------------- An optimist believes that we live in the best of all possible worlds. A pessimist fears this is true.
} I have projected stereo images successfully by creating red/green anaglyphs } on transparency material. We took our digital images, combined them using } PhotoShop or similar program, and printed them on a dye-sublimation } printer. Using a standard overhead projector and the red/green glasses, } the images greatly impressed my audience. The dual projector system seems } a bit specialized. } } Henk
The Anaglyph system is indeed simple and easy to use and requires only an ordinary projector and screen but it does have at least two snags. Apart from the opportunities that it gives to "Murphy" to attack when two exposures must be recorded on the same piece of film in register and through different colored filters, many people get very uncomfortable when the color of the image presented to one eye is very different from that presented to the other. The phenomenon is referred to as "color bombardment" and was much discussed by Vernon Barber in the late seventies/early eighties with reference to Scanning Electron Microscopy.
The second snag is that you cannot show colored images using this system. For instance you cannot use it to show stereo views of confocal data in which you have superimposed separate 3D images obtained from 2 or 3 different fluorescent dyes in the same specimen. Likewise, you cannot show stereo SEM images in which the SE signal is recorded in tones of grey while the BSE image is superimposed in shades of red.
In addition, although there is usually less overlap between the red and blue filters than between the red and the green, the light output in the blue from most projectors is insufficient to convey many grey levels to the screen and back.
Once you accept the difficulty of finding an "aluminized" screen, the Pol system has much to recommend it.
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
I have done just as you suggest. Using a portable silver lenticular (sp?) screen (~$200.00) which does not depolarize the images, a beautiful 3D image slide show can be projected.
In our department, we have had the capability of mixing 2D and 3D images in a single slide presentation for about 5 years now and I have even done a few 3D demonstrations at a local high school -- with great success (held their attention from start to finish)!
As you have outlined, two matched projectors are required as well as polaroid transparancy sheets and glasses. Using a single piece of Scotch Tape to attach (hang) a piece of the polaroid transparancy over the front of the objective lens of each slide projector (each piece oriented 90 degrees to the other) is sufficient to project a polarized image on the screen.
The glasses, which each person viewing the projected images must have, represents a bit of work. I made a template for the eyes (no ear supports/hangers needed - see below dowell sticks) and traced the template image for as many glasses as I needed on firm, but flexible, cardboard that could be cut with a good pair of sharp tipped sissors.
After tracing and cutting the cardboard, using carefully applied small pieces of Scotch Tape, I taped the polaroid 'lens pieces' from the same sheet of polaroid material used in front of the projector lenses [CORRECTLY ORIENTED] for left and right eye. By correctly oriented, I referring to the necessity of having all glasses with the angular orientation of the polaroid pieces identical -- all left and right eye orientation must be the same in the glasses and matched to the orientation of the pieces in front of the projector lenses. This way, all observers receive the same (correct) image in the left and right eye.
When the 'lenses' were all taped in place, I then taped these glasses by their right or left edge (makes no difference) to a 12 inch length of 1/4 inch wooden dowell stick so that viewers could hold the glasses in front of their eyes (and personal prescription glasses if that's the case) much like holding those small masks at a masquerade party. This may seem a little amusing, and sometimes gets a laugh at the beginning of a 3D presentation, but it was the method I developed to circumvent any health hazard concerns from the repeated public use these glasses -- this is why there are no ear supports as on regular glasses.
From the SEM image standpoint, I take two photos of the same subject (carefully aligned to be nearly the same field) at angles of +/- 5 degrees off verticle by stage tilts. I can then make direct positive slide transparancies with B/W 35 mm microfilm for projection. Both images are matched for size when projected on the screen and simply superimposed -- the glasses and polaroid pieces do all the separation of images to create a very clear and satisfactory 3D image for the viewers.
Hope this helps. This note has been a sort of 'stream of thought' description so please don't hesitate to ask any questions.
} We are encountering problems in disposal of uranium salts as our site } safety officer considers it as a mixed/radioactive waste. I wonder } what is the experience of other microscopists regarding } radioactivity/disposal of uranium compounds. } Dear Manoj, The law in New York allows us to pour uranyl acetate down the regular sink for the amounts used for staining. For larger amounts, they have speci- fied procedures, and our responsibility is to give the material to them with the activity listed on a data sheet--they do the rest. Yours, Bill Tivol
In response to you questions regarding the use of a GATAN disc grinder on diamond lapping films:
While I don't mean for this response to be an advertisement for product, I must preface my remarks by mentioning that the experiences and methods described here occurred using the products of my employer: BUEHLER, LTD in our development laboratory. Therefore, I, like Mr. Henriks, do have a commercial interest in your successful specimen preparation.
A1: In response to your query on experiences with GATAN's disc grinder on Diamond Lapping Films, I also have not used the GATAN fixture; however, I have tried (successfully, I might add) to grind metallurgically mounted samples (1-1/4" in diameter) on BUEHLER's ULTRA-PREP diamond films. The problem of adhesion between the sample and the film, when water is used as a lubricant, does cause movement of the diamond film on the glass wheel. Our experiences indicate that wheel speed makes no difference. However, success was achieved by using a PSA (Pressure Sensitive Adhesive) backed film, attached to the surface of a POLIMET Pad. It was determined that the diamond films are extremely flat compared to comparably sized silicon carbide papers, and therefore a thin film of water between sample and abrasive film causes the sticking problem. Breaking up this film of water is the key to stopping the sticking. The POLIMET pad is perforated, leaving small depressions in spots under the film, while maintaining the rigidity needed to keep the sample flat. Another option for you is to break up the smooth, flat surface in contact with the film; i.e. the bottom of your GATAN fixture (this may not be the more convenient option, however).
With regard to your question about the DELRIN feet of tripodding type polishers, I have had experience here as well. BUEHLER's TRIPOINT POLISHER(TM) also uses DELRIN feet. These DO wear when held against the diamond films. However, the purpose of the TRIPOINT POLISHER(TM) type fixtures was originally to cross-section microelectronic materials. While the DELRIN does abrade slowly, it does not abrade anywhere near the rate of silicon, GaAs, etc. The micrometers on which the feet are mounted are there for the purpose of adjusting tilt and pitch of the sample, as well as adjusting for the insignificant (when polishing most materials) wear encountered on the feet.
The influence of the DELRIN foot area is important with regard to the sticking problem. Obviously, the smaller the feet, the less sticking that will occur. However, another variable should be considered: The abrasive size. On a 30 micron diamond film, the TRIPOINT POLISHER(TM) does not stick because the diamond is large enough to allow air to break up the water film between foot and diamond film. However, when you get in the smaller sizes, e.g. 1 to 0.5 microns, the DELRIN feet become 'polished', and the sticking problem begins to increase. This is sometimes felt as a steadily growing chattering of sample on abrasive surface. My suggestion is to cut grooves in the DELRIN feet in order to break up the large surface area found there.
A2: The 3X rule of abrasive damage removal was proposed by a competing metallurgical supply company a number of years ago. It is a very good guideline to work with, but it is not a scientifically proven rule, as far as I am aware. Of course ductile materials will tend to follow the rule more than hard, tough materials. Therefore, I don't see any reason why you must use every abrasive size while grinding your materials. However, for samples where edge retention is important, or where a cross-section of extremely thin layers is being attempted, I have found that the prescribed steps of 30, 15, 6, 3, 1 and 0.5 micron diamond films are necessary. Especially the latter case.
If you would like to eliminate a significant portion of the grinding that is necessary, you may want to try a diamond wafering blade which contains a finer abrasive than the 68 micron diamond you are currently using. We offer a blade which contains diamond about 1/13th this size, and which is designed for exactly this purpose. We call this the L5 series blade. You can contact me directly by e-mail, phone or fax for more info. The surface finish produced is close to that produced by a 3 micron abrasive film. I do have a BUEHLER produced, technical brochure (Note: contains info on only our products) which describes the effects of cutting force, speed, abrasive size, etc., if you would be interested.
A3: Our experience has been that low loads give better results on diamond lapping films than high loads. This is especially true with ductile materials. Cleaning the films with a fresh, folded paper towel during the grinding step also improves the result obtained on most materials. The towel captures loose diamond and swarf before it can be reintroduced to the sample; causing scratches or embedding. Make sure to use a fresh towel with every abrasive size change.
A4: I differ with Mr. Henriks on this question. I don't suggest rotating the specimen during each grinding step. Rotating the specimen does not introduce fresh abrasive to the sample. Only the wheel rotation does this.
When the scratch pattern from the previous abrasive is removed, the damage may or may not be removed (again, depending on the material). However, this may not be relevant in the case of ceramics such as the silicon nitride you mentioned. This is because the damage mechanisms are different between ductile and hard/tough materials.
A5: I do agree with Mr. Henriks on this question. I was, at one time, Application Engineer at SBT, and have had the opportunity to work with all of their carbide footed grinding fixtures. The free-floating nature of their fixtures does reduce the load on the specimen. I believe they still offer a fixture with spring counterbalancing, which further reduces the specimen load. These fixtures have an adjustable lock-nut which is dialed to a relative depth. This nut contacts the piston housing when the piston reaches it's lowest point. This stops the polishing.
A6: I am not familiar enough with the GATAN fixture to answer this question fully. However, my experience with using a tripod type device (as Mr. Henriks suggested) with the sample mounted in the middle of the fixture produces a sample damaging instability during polishing. Shifting of the fixture from a base created by the sample and two DELRIN feet to the sample and another set of feet causes edge chipping if not carefully monitored.
If I can answer any questions for you regarding diamond lapping films, fixturing, etc., please don't hesitate to contact me directly.
Best regards,
Scott D. Holt BUEHLER, LTD. 41 Waukegan Rd. Lake Bluff, IL 60044 Phone: (847)295-4546 Fax: (847)295-7942
Here are my respones R1-6 to your comments C1-6 from my answers A1-6 to your questions Q1-6:
Response 1: As you may have read in the response from Scott Walck:
"If you use the diamond films with the GDG, when the sample becomes planar with the base of the grinder, then the grinder tends to stick to the film by surface tension because of the large area. This is even more of a problem at the lower grit sizes."
This will not happen on a soft cloth with diamond spray because you are not able to form a vacuum as you can with our film. The downside is that you will end up with rounded edges on your sample. The vacuum problem is exacerbated by using the small increments which are best to use with the gatan Disc Grinder. As you polish the sample down or use very small increments, the entire base becomes nearly co-planar which causes the vacuum to form. On the SBt fixtures, there is a space between the outside ring and the center piston which minimizes this contact area and hence the vacuum effect.
I am not surprised that Gatan recommends 3M films, they are not too likely to refer their customers to SBT! As far as the 50u increments go, that may be an advisable number to use where it minimizes rocking on the sample caused by the sample sticking out below the bottom of the fixture, but also may minimize the vacuum effect. I'm guessing that is the reason.
RE: The Tripod Polisher (R) The sof t delrin feet of the Tripod Polisher are actually made to wear out. The idea is that you don't want any material which is harder than your sample to make contact with the wheel. We use this logic in Tripod Polishing because the sample becomes very thin and fragile and we do not want any particles from WC-Co feet to containate the film. Furthermore, the Tripod Polisher (R) has the facility to adjust for the change in level of the feet. The fixtures with the tungsten carbide do not have the ability to be adjusted after polishing has begun so we try to maintain the parallelism as best we can by resisting wear. The measurements on the micrometer are, for the most part, not used. You make "relative" adjustments with the micrometers, but you do not actually measure your sample thickness with them. Sample thickness is typically determined optically.
The delrin feet on the Tripod Polisher (R) can also create a vacuum if the are polished flat and co-planar. In fact, we sometimes grind facets onto the feet to maintain more of a point contact with the film. One of our competitors sells a tripod Polisher with "non-rotating" micrometers. They charge you more for them and they are counterproductive. By utilizing non-rotating micrometers you are creating the very situation you are trying to avoid.
Response 2: If I understand this now, you are cutting your sample with 68u diamonds which would indicate a damage layer of 3 x 68 or 204u. I suppose you would need a fairly thick sample if you need to cut on both sides of the sample. I'm not sure how to avoid this. Of course, this 3x rule is a general rule and will vary with sample type. Perhaps you would like to do a little study for us in your spare time?! :} ) Assuming that you are looking for a 1u final thickness and that you will use each of these steps, the process would then be something like this:
30u: Polish until 105u thick 15u: Polish until 51u thick 6u: Polish until 21u thick 3u: Polish until 10u thick 1u: Polish until 4u thick 0.5u: Polish until 2u thick Colloidal silica for final polish.
The rough formula I use is (3 x current particle size) + particle size of next film in sequence. This gives me the amount of material I need remaining after a particular step in order to remove thedamage and still have "good" material to work with. Of course, you may want to be a little more conservative or aggressive depending on your confidence level. You can remove film steps and apply the same formula to come up with alternative sequences.
Response 3: You got me on this one. Actually, you probably got me on the other ones too, but at least I could come up with some reasonable answer for you! : {)
Response 4: No. The SBT polishing fixtures have specimen mounts that are maintain in a single orientation by way of 2 locating pins that come out of the center piston and fit into holes in the back of the removable mouting block. The piston does not rotate because there is a slot in the piston. A set screw then passes through the outside ring and into the slot to prevent rotation of the piston relative to the outside ring.
Response 5: We have a micrometer type arrangement at the top of the piston which allows you to dial in an amount of material to remove. This is done after a simple zeroing procedure. The dial at the top of the piston will stop the polishing process at the desired thickness by making physical contact with the outside ring. This prevents the piston was moving down any further.
Response 6: The Tripod Polisher is extremely effective in polishing very flat and parralel samples. You have great ability to make adjustments to correct for any errors in sample mounting and other inaccuracies in the process. However, since you have the ability to make the changes, you generally need to make them which makes the process a bit more cumbersome. You can definitely get a great sample, but you do have to work for it.
Our other fixtures rely more heavily on minimizing the variables. We have a mounting press of sorts that you use to firmly press your sample against your mounting block minimizing glue thickness and maximizing parallelism. We also recommend that you polish your mounting block while mounted in the fixture prior to mounting your sample. This ensures that your moutning block is parallel to the plane of the tungsten carbide feet. As there are no adjustments possible, it is best to follow the entire procedure carefully. Even with no adjustments, you can get a pretty darn parallel sample! Wealso make a polishing machine (Model 920) that can be fitted with workstations to hold and rotate the fixtures. We also build custom attachments (for less than you might think!) which allow a cusomer to polish 2 sides parallel to each other by simply flipping the sample over and not re-mounting.
RE: Cyanoacrylate My understanding is that any cyanoacrylate will be acetone soluble. Perhaps they just don't list acetone on the label for some reason? A sure bet is to get something like a nail bonding glue that is used to fix fingernails or attach fake nails. Check the cosmetic counter. I will try to put a tube of super glue in the mail to you, but I am not certain if it will still be good when it arrives. I have tried this before and had problems. That's the reason we stopped supplying it - it seemed a little silly to make an unhappy customer because of a $3 bottle of glue that we don't even make!
Well I think I am worn out. You have asked a lot of very good questions and it is obvious that you take your sample preparation very seriously. I am always pleased to help in any way i can and i encourage you to contact me whenever i may be of assistance. Of course, next time i wouldn't mind if talked about your application for some South bay Technology equipment! : {)
Best regards-
P.S. I would love to get a copy of other responses you get concerning this topic. Thank you!
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com sbt-at-msa.microscopy.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
I am getting ready to purchase a replacement sputter coater and I was hoping to sollicit user comments (either good or bad or general) regarding either the Pellco SC-7 Automatic sputter coater or the EMITech K550 automatic sputter coater (aka EMS 550) and/or the integration of the thickness monitors available for each unit.
PLEASE EMAIL ME DIRECTLY !!!!
ALL COMMENTS WILL BE KEPT PRIVATE !!!!
(I am not looking to publically condemn nor praise either instrument - I like both companies, but without having worked with either instrument, I'm looking for comments from those out there who have.)
Thank you in advance.
Richard E. Edelmann Electron Microscopy Facility Supervisor 352 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513-529-5712 Fax: 513-529-4243 E-mail: edelmare-at-muohio.edu
} We would like to know if anyone has an opinion on the effectiveness of } using a silver screen and double projectors for 3D imaging. We do not want } to commit ourselves to buying a screen (which is quite expensive) and then } find it has been a waste of money. } } We would like to hear from anyone who has tried this or any other } alternative method of displaying 3D images to groups of people.
Dear Mark, We have two large screens--they are aluminum, but look silvery; I assume these are what you want--for 3D display with polarized light & dual projectors. The images are quite spectacular and always make a big impres- sion. If you have many 3D images to display, this kind of system may be well worth the initial investment. The screens last for a very long time; ours are } ~25 years old and are as good as ever. I have no idea of the current price for these screens. Yours, Bill Tivol
We are encountering problems in disposal of uranium salts as our site safety officer considers it as a mixed/radioactive waste. I wonder what is the experience of other microscopists regarding radioactivity/disposal of uranium compounds.
Manoj MISRA Unilever Research US Edgewater NJ
Here in Bristol, we are, as long as the University Safety Office knows, and keeps some control over it, allowed to dispose of up to 100g of uranium per week (I think it is) in with the normal office/laboratory rubbish. I was surprised when I first found this out, but it has allowed us to dispose of a collection of unwanted uranium oxides and the like. Keith
I am looking for suppliers of SEM cryopreparation and cryo-transfer systems. Vendors, suppliers or colleagues with e-mail or fax numbers please respond directly to me. And yes, I have done the obligatory netsearch with very few direct scores.
Terri- A bain marie *is* a double boiler. It was invented by a medieval Jewish alchemist known as Mary the Prophetess, hence the name. Aren't you glad you asked? Steven Slap 102134,1660-at-compuserve.com
Does anyone know a good reference on the possible mechanisms of fluorescence quenching? Especially processes which involve the interaction of fluorophores with transition metal complexes or colloidal particles, from a chemistry perspective (electron transfer, molecular orbitals and so on).
I was wondering whether any of you could help giving me some tips or references where to look for counting strategy for particles (such as number of particles to be counted regarding the size and/or magnification). At this moment I'm trying to obtain polystherene particles size distributions by TEM and SEM.
Has anyone experience in working with SIGMA SCAN PRO, made by Handel, for such a distribution? and for images analysis? I would like to hear any comment about it or about any other software suggested for the subject.
Thanks a lot. Silvia Montoro CERIDE - Centre for Research and Development Santa Fe - Argentina csedax-at-arcride.edu.ar
Message-Id: {199602081547.JAA26446-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
At 07:39 PM 2/7/96 -0600, you wrote:
.....Do you have any info, printed or otherwise, regarding your pricing structure?
*************** No, I don't have anything formalized. I just sat down and calculated what we were spending to get one case done (costs, including salaries, supplies, overhead, etc., divided by total number of micrographs, and then how many micrographs were shot per case). To that I add what I think is reasonable to support equipment replacement. That's what I charge. For clinical cases the department adds a professional component.
I have a question on etching polysilicon on SOI material. I need to find an etch that will etch the polysilicon and not the silicon on the buried oxide. Feel free to call me or send your phone number. Thanks, Mark Windland Honeywell 612-954-2845
We followed the same procedure as Henk (using PhotoShop to produce red/green anaglyphs from pixel shifted z-sections) but then digitized the images onto slide film (Kodak Elite) using a Polaroid Palette. A single projector then did a great job (in fact it actually worked with red/blue glasses...)
Hank Colijn wrote: I have projected stereo images successfully by creating red/green anaglyphs on transparency material. We took our digital images, combined them using PhotoShop or similar program, and printed them on a dye-sublimation printer. Using a standard overhead projector and the red/green glasses, the images greatly impressed my audience. The dual projector system seems a bit specialized.
Henk
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
A few comments on this thread from our experience:
We have mounted on polarizers on the ends of cardboard tubes. The tubes are approximately the diameter of the Kodak projector lens barrel. We put a lenght of the thin packing sponge material that our hazzardous chemicals come wrapped in around the inside of the tube to provide friction and hold the tube in place. This allows the polarizers to be rotated to get maximum extinction with the glasses. We order bulk glasses for a nominal cost. Most come with the polarizers set at 45 degrees to the horizon. However, some glasses come with the polarizers horizontal and vertical. The tube system allows us to adjust or projector polarizers to match either set of glasses.
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
} Date: Wed, 07 Feb 1996 16:31:59 -0500 } From: zaluzec-at-microscopy.com (Nestor J. Zaluzec) } Subject: Minor Problems } X-Sender: zaluzec-at-microscopy.com (Unverified) } To: Microscopy-at-Sparc5.Microscopy.Com } } G'day Subscribers... } } We've got a minor glitch in the subscribe/unsubscribe system. } Unfortunately, I'm on vacation and too far away to fix the link. } This just means there will be a delay if your trying to } unsubscribe. I'll be logging in remotely and doing manual } updates, but the connections are sometimes slow from here. } } I'll be back in town the middle of next week and we should } be back to normal. } } Sorry for the inconvenience } } Nestor } Your Friendly Neighborhood SysOp } } Weather report: 34 C, partly cloudy, and the view on } the beach is great. ;-) } }
Vacation? VACATION?! That's pretty cheeky. And 34C? Last week we had MINUS 34 C! Enjoy your break, Nestor.
After reading a few of the messages on this server from users in different states representing various regions of the USA, I think about the growing pains we are facing with what it seems "inconsistent regulation in the UNITED" deserves a revisit? Two items are worth recollecting: osmium and uranyl disposal. If you think that the disposal of these chemical (which by all means are hazardous) think about the follwing. During one my visits to California (NASA AMES RESEARCH CENTER) I was prohibited (and instructed so formally) from pouring SALINE (That's right PBS!) down the drain! Thus, I immediately asked where I should urinate (you know amonia, amino acids, etc). Even though the biosafety officer did not have a good answer (take that back... regulations) he was rather upset. The moral of the story: I learn quickly after arriving to the states long ago, that -the outcome of a procedure matters less than the implementation of the regulation that requires it-. That is the good news, the bad one is thatit will get worse if we do not inject a tiny bit of common sense into all of this mess and others. My two daughters were taught at school already that animals are terribly mistreated by industry and researchers, but I had to point out to her the reality behind the production of a juicy burger and/or taste fried chicken nudget? This was easy for me because when I was her age I butchered the chickens we ate and witness the killing and butchering of the pigs, cows, etc. Go on, make your contribution, and get started now-tell as really is!
********************************************************************* *Cesar D. Fermin, Ph.D \|T|/ Fax (504) 587-7389 * *Tulane Medical School /|M|\ Voice Mail (504) 584-2618 * *Pathology/SL79 \|C|/ Secretary (504) 584-2436 * *New Orleans La 70112-2699 /|*|\ Lab/Techn. (504) 584-2521 * * {Fermin-at-tmc.tulane.edu} ----} Prof. Pathology & Otolaryngology * *http://www1.omi.tulane.edu/departments/pathology/fermin/cdftop.html* *********************************************************************
A visitor to our lab from Switzerland brought us an adhesive used for sealing wounds, called HISTOACRYL Blue. We have found it to be very useful and would like to find a supplier in the US. Any leads will be appreciated.
This question was asked before, and I replied direct to the poster not to the listserver. Your supplier for polyclonal nitric oxide synthetase antibody is:
AFFINITY BIOREAGENTS, INC 14818 West 6th Avenue, #13A Golden, CO 80401
We have just received the new (1996) Lindscott's Directory of Immunological and Biological reagents (Order from Lindscott's Directory, 4877 Grange Road, Santa Rosa, CA 95404; phone (707) 544-9555, Fax (415) 389-6025) which is a good place to start looking for antibodies. Some entries for anti-nitric oxide synthetase antibodies:
POLYCLONALS:
Rabbit antibody against inducible NOS, mouse macrophage:
Alexis Corp. P.O. Box 927190, San Diego, CA 92192, USA Tel: (619) 658-0065, Fax (619) 658-9224
Rabbit antibody against inducible, neuronal or endothelial NOS:
Oxford Biomedical Research P.O. Box 522, Oxford, MI 48371 Tel (in US) 800-692-4633, Fax (810) 852-4466
MONOCLONALS:
IgG2A against brain NOS, against mac, inducible NOS, against NOS EC:
Transduction Laboratories 133 Venture Crescent #5, Lexington, KY 40510, USA Tel (606) 259-1550, Fax (606) 259-1413
Against inducible NOS:
Research and Diagnostic antibodies P.O. Box 8300, Berkeley, CA 94707 Tel (510) 262-9000, Fax (510) 262-9127
Since I've never used these I can't tell you whether they work, but they are probably good places to start.
Richard D. Powell ************************************************************* NANOPROBES, Incorporated 25 East Loop Road, Suite 124, Stony Brook, NY 11790-3350, USA http://www.lihti.org/research/ecodev/incubten/nano/home.html
} } Does anyone in cyberspace know where I can obtain and antibody to Nitric } } Oxide Synthase which will work on mouse brain. } } } } Dr Terry Robertson } } Electron Microscopist } } Department of Pathology } } University of Western Australia } } Nedlands 6009 } } } } phone 346 2935 } } Fax 346 2891 } } email troberts-at-eosin.path.uwa.edu.au }
(If this appears twice, please ignore the earlier version-my mistake for E-mailing Microscopy-at-Sparc5 instead of MSA. My apologies)
Dear Christine Block:
Someone else asked about antibodies to nitric oxide synthetase, and I replied direct to the poster not to the listserver. Affinity Bioreagents supplies a polyclonal antibody to nitric oxide synthetase; their address is:
AFFINITY BIOREAGENTS, INC 14818 West 6th Avenue, #13A Golden, CO 80401
For other polyclonal and monoclonal antibodies (in reply to the earlier question):
We have just received the new (1996) Lindscott's Directory of Immunological and Biological reagents (Order from Lindscott's Directory, 4877 Grange Road, Santa Rosa, CA 95404; phone (707) 544-9555, Fax (415) 389-6025) which is a good place to start looking for antibodies. Some entries for anti-nitric oxide synthetase antibodies:
POLYCLONALS:
Rabbit antibody against inducible NOS, mouse macrophage:
Alexis Corp. P.O. Box 927190, San Diego, CA 92192, USA Tel: (619) 658-0065, Fax (619) 658-9224
Rabbit antibody against inducible, neuronal or endothelial NOS:
Oxford Biomedical Research P.O. Box 522, Oxford, MI 48371 Tel (in US) 800-692-4633, Fax (810) 852-4466
MONOCLONALS:
IgG2A against brain NOS, against mac, inducible NOS, against NOS EC:
Transduction Laboratories 133 Venture Crescent #5, Lexington, KY 40510, USA Tel (606) 259-1550, Fax (606) 259-1413
Against inducible NOS:
Research and Diagnostic antibodies P.O. Box 8300, Berkeley, CA 94707 Tel (510) 262-9000, Fax (510) 262-9127
Since I've never used these I can't tell you whether they work, but they are probably good places to start.
Richard D. Powell ************************************************************* NANOPROBES, Incorporated 25 East Loop Road, Suite 124, Stony Brook, NY 11790-3350, USA http://www.lihti.org/research/ecodev/incubten/nano/home.html
} } Does anyone in cyberspace know where I can obtain and antibody to Nitric } } Oxide Synthase which will work on mouse brain. } } } } Dr Terry Robertson } } Electron Microscopist } } Department of Pathology } } University of Western Australia } } Nedlands 6009 } } } } phone 346 2935 } } Fax 346 2891 } } email troberts-at-eosin.path.uwa.edu.au }
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The Materials Analysis group at SEMATECH in Austin, TX has an immediate opening for a TEM lab Technician. SEMATECH is a government/industry consortium working on advanced projects related to the fabrication of 0.25/0.35um CMOS integrated circuits.
For more information please contact Carolyn Gondran -at- 512-356-3149 , Philippe Maillot -at-512-356-3944 or Ted Boden 512-356-3324 or respond by direct e-mail.
JOB DESCRIPTION: Sample preparation of semiconductor materials and devices, both plan view and cross-section, for TEM. Develop improved methods of sample preparation. TEM operation to facilitate this development will be encouraged. Photographic processing and filing of TEM negatives and prints. Maintenance of sample preparation and dark room equipment and supplies
JOB REQUIREMENTS: 2+ years experience in TEM sample preparation and photographic processing required. Familiarity with tripod, dimpling and FIB techniques desired. This position requires strong organizational skills, and in particular, the ability to work on multiple tasks while maintaining precise records and tracking procedure. Experience in microelectronics industry a plus.
Associate degree in physics, chemistry or related subject.
Registered-mail-reply-requested-by: Ronald.Cohn-at-SYNTEX.COM MR-Received: by mta RVAX.MUAS; Relayed; Thu, 08 Feb 1996 17:25:49 -0800 MR-Received: by mta RVAX4; Relayed; Thu, 08 Feb 1996 17:25:49 -0800 MR-Received: by mta CHUB1; Relayed; Thu, 08 Feb 1996 17:20:30 -0800 Disclose-recipients: prohibited
List subscribers,
We need to identify apoptotic cells in cultures of cells grown in suspension. We would like to do this using the TUNEL (terminal transferase-mediated dUTP-biotin nick end labeling) technique with either fluoresence or enzyme-substrate reaction product as the read-out. I would appreciate hearing from anyone who has used this technique regarding which commercial labeling kits may be better than others, recommended protocols, etc. Thanks in advance.
Ron Cohn Structural Biology Laboratory Roche Bioscience Palo Alto, CA ronald.cohn-at-syntex.com
On March 8, 1996, a workshop on confocal microscopy will be held in the Laboratory of Physiology, KULeuven, Belgium.
Program:
08.30: Registration 09.00: Welcome: Prof. Dr. J. Janssens, Dean of the Faculty of Medicin KU Leuven 09.15: Confocal vs conventional microscopy. H. van der Voort, J. Bauman, H. Vrolijk, W. Sloos & H. Tanke, SVI Hilversum, UA & RU Leiden, The Netherlands 09.45: Spontaneously spreading calcium waves in rat cardiac myocytes: implications of the data acquisition by confocal microscopy. W. Wussling, Martin Luther University, Halle, Germany 10.15: Coffee and demonstrations on confocal microscopes 11.00: Confocal laser scanning in the reflection mode. Z. Mischal, CNRS, Villejuif, France 11.30: Free communications 12.00: Lunch 13.15: Demonstrations on confocal microscopes 14.00: Confocal fluorescence life time imaging with two-photon excitation. H. Gerritsen, J. Sytsma & J. Vroom. Universiteit Utrecht, The Netherlands 14.30: Manipulation, N-dimensional visualisation and measurement of the living cell with super-resolution and super-contrast. P. Van Oostveldt, U Gent, Belgium 15.00: Coffee and demonstrations on confocal microscopes 15.45: Free communications 17.00: Final conclusions and farewell. B. Himpens, KU Leuven
Date and location: The workshop will be held on Friday, March 8th, 1996 at the KULeuven Lab Physiology, 8th floor Onderwijs & Navorsing, Gasthuisberg Herestraat 49 B - 3000 Leuven Belgium
Contact person: B. Himpens Lab Physiology Herestraat 49 B - 3000 Leuven tel + 32 16 34 57 27 or + 32 16 34 71 46 fax + 32 16 34 59 91 E-mail: Bernard.Himpens-at-med.kuleuven.ac.be
More information and online registration: See URL: http://www.kuleuven.ac.be/kuleuven/news/wcm/
Peter Stalmans Peter.Stalmans-at-med.kuleuven.ac.be Laboratory of Physiology KULeuven Herestraat 49 B-3000 Leuven Belgium tel: +32-16-34 71 46 fax: +32-16-34 59 91
Dear All, A few of you have expressed surprise at my comment on disposal of active waste here in Bristol. I thought (though there is no direct microcsopy content - shouldn't this be in one of the safety groups??) you might like a couple of quotes from the relevant University safety handbook. "Aqueous waste. Our primary waste disposal route is via the sink... This is both the least restrictive and the least environmentally damaging" It then goes on about how this would be diluted by all the other waste water the University discharges into the sewars. "Very low levels of solid radioactive waste disposal are permitted by out authorisation from HMIP (Her Majesty's Inspectors of Pollution) to the normal waste bins.""No visible radioactive signs are to be present on the box or bag used for disposal and please try to ensure that the package does not look interesting..." There are records that have to be kept, and regulations on there being no external contamination, maximum permitted levels, only one lot per waste bin, etc.
-- Dr. Keith R. Hallam University of Bristol, Interface Analysis Centre, Oldbury House, 121, St. Michael's Hill, Bristol, BS2 8BS, England Telephone: + 44 (0)117 925 5666 | E-mail: k.r.hallam-at-bristol.ac.uk Facsimile: + 44 (0)117 925 5646 | URL: http://zeus.bris.ac.uk/~phkrh/
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Does anyone know of some "good" PC bases image processing software designed for microscopy applications? I need to do some time based studies on fluorescent DNA strands.
I have an optical trap set up to hold and maneuver the strands as attached to sub micron latex beads. I want to study the effects of various enzymes and environments on the DNA while in the trap.
Any suggestions as to software and its required (desired) hardware would be appreciated.
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Does anyone know of custom or commercially available UV microscope objectives for use in Laser Induced Fluorescence studies?
I have contacted Zeiss, Nikon, Olympus and Leica. Zeiss has the best of the group but the UltraFluars all fluoresce in my UV laser beam (266 nm). I want as high an NA as possible, i.e. immersed optics, so the reflective designs, e.g. Ealing, I have seen are not fast enough.
Fluorescence excitation wavelengths are tunable between 260 and 290 nm. Fluorescence emissions are from 300 to 500 nm.
Any suggestions would be appreciated. I can afford a custom design after I prove the concept at more limited wavelengths.
Any comments from cyberspace on the additives that EM supply houses sell to add to epoxy resins that are supposed to improve sectioning properties? I am referring to products such as Poly Cut Ease (Polysciences) or SureCut (EMS). Do people think they actually help? Are there any disadvantages?
Thomas E. Phillips, Ph.D. Associate Professor of Biological Sciences Director, Molecular Cytology Core Facility 3 Tucker Hall University of Missouri Columbia, MO 65211 (314)-882-4712 (voice) (314)-882-0123 (fax)
With regard to using 3D aluminised screens for projecting polarised images , I can only add in support of Jim Pawley's comments on the advantages for imaging colour - in my case cell reconstructions with colour coded organelles - which come out only with the polarised system.
As regards sourcing a screen try your Chemistry colleagues who probably have one lurking somewhere as they may project molecular graphics in this way for teaching. This is where I borrow my portable screen from here in Glasgow !
Laurence Tetley Dr Laurence Tetley IBLS EM Centre Joseph Black Building University of Glasgow Glasgow G12 8QQ
I am searching for the most comprehensive articles about critical point drying techniques. All suggestions are appreciated.
Any problems with going directly from ethanol into carbon dioxide or is amyl acetate useful in between? My specimens look OK using just ethanol.
Also: I am experiencing a run of failing (most often between 400 and 800 psi) front window Dowty seals on my drying unit even if I do only 'dry' (carbon dioxide only) test runs. Is anyone else having this problem? This device has been operating for 17 years with few leaks. All sealing surfaces are smooth and clean.
Thank you in advance.
Jim Romanow Electron Microscopy Facility Physiology and Neurobiogy Department The University Of Connecticut Storrs bsgphy3-at-uconnvm.uconn.edu
I have had over a dozen requests for this information so I am posting it directly to the list:
We have always purchased our stereo glasses from Ted Pella 1-800-237-3526 outside of California They are in cardboard mounts and relatively inexpensive. However, I did not see them listed in the current catalog. I hope they still carry them. Any one know of an alternate supplier?
Jay Jerome ************************************************************** * aka: W. Gray Jerome * * Dept. of Pathology * * Bowman Gray School of Medicine of Wake Forest University * * Medical Center Blvd * * Winston-Salem, NC 27157-1092 * * 910-716-4972 * * jjerome-at-bgsm.edu * **************************************************************
I have prototype LVSEM that employs the stage mechanism from a Philips EM 430 TEM. I would like to purchase a cold stage for this instrument if I can find one (standard Philips rod). Anyone out there about to "decommission" one?
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
Hello Everyone, I'd just like to put in my two cents on the subject of multiple users of an SEM (or in my case an electron microprobe)
Our instrument is used at least 16 hours a day and since I'm only paid for eight almost all our users are trained to use the machine independantly. This allows them faster access to the machine and saves them or their advisors money. The only people we don't encourage to become independant users are those who will only use the machine once or twice. Most users require 3-4 daytime shifts before i will let them use the machine independantly (I give them my home phone number too).
In the six years I've been running the lab I can't recall one incident where the machine has actually been damaged by inexperienced users. On several occasions it has been temporarily put out of commission (computer problems, etc) but usually I can get it back on line immediately the next day. Although there is the potential for a user to damage the instrument (i.e during a sample change) most of the rest of the instrument is fairly fool proof. It is easy to screw up your analyses, but difficult to hurt the machine.
I'm sure the machine would last longer and require less service if I was the only one operating it, but we don't have that option. I also feel that users are much better off acquiring their own data, they know what they want and can change their strategy if they find something unexpected
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Dear Bonnie, I run a Materials Engineering EM lab at the University of British Columbia, with two SEMs and a TEM. Most of the researchers who use the instruments are graduate students and Research Engineers. I have always encouraged these people to do their own research on the SEMs, as much as possible, since only they know exactly what they want and that way one me can keep three instruments optimally used. After I show them how to use the SEM, I watch and encourage them to ask me for further instruction for picture taking, higher mag, etc. Some people, who have a lot of work to do, may gain my permission to use the SEM after hours, but only after I feel they have gained a lot of experience during working hours, when I am there to supervise, and only after I have instructed them on how to properly shut down and what to do in the event of problems. I also purchased the instruments originally with ease-of-use in mind. They are fully automatic and easy to get good results on. I must feel that the person has a good understanding of the important issues and knows how to properly handle the instrument. Mind you, I don't have a field emission SEM. I have had damage, but only once in 15 years and it could have happened in normal hours.. It also helps to scare them, be "the ogre of the EM lab". Mind you, we are a teaching lab, so the learning is as important as the doing. I know many EM operators do not like the idea of letting any ham-fisted graduate student or engineer loose on their instrument, but modern SEMs really can be operated by any intelligent being. So long as you satisfy yourself as to this person's knowledge, experience and caring, I'd see no harm. Hope this helps.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
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At 10:17 AM 2/8/96 EST, you wrote:
".....to form yellow, iridescent hexagonal prisms of lead iodide....."
I stained one grid with our lead stain alone, and another with our uranyl acetate stain alone. The lead stained grid had the hexagonal deposits and the other did not. So it's not the uranyl acetate. It's interesting that lead iodide forms hexagonal crystals. *******************
".....Perhaps these are your crystals - a bit of a long shot since you did not mention iodine in our elemental analysis of the crystals...."
No, there is no iodine in either of our stains, or any of the sample preparation steps; and I did not get an iodine peak with the microprobe. *******************
I have borrowed an anti static gun like the Zerostatic Eliminator that you suggest. We are in the process of trying it out. At least the grids don't jump up and cling to the lid of the plastic petri dish any more.
These things help a great deal when using a glass knife. We use lecithin in the epoxy accelerator . ( use eual weight of lecithin and accelerator, then double the volume for use) A glass knife will last a long time if you have it in the resin. It was a great help to students just learning to section, They could get usable stuff the first day at the microtome. It can give a blotchy look to empty resin areas or those that have little density, but most tissues are "busy " enough that you never notice it. See Mollenhauer , 1986, J. EM Tech 3:217-222 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
I'd like some input, opinions etc. on the pitfalls of having multiple operators on an SEM. My situation is that I am part of a materials analysis group that provides analytical services to corporate technology. Our group consists of a number of engineers and technicians with expertise in a variety of analytical areas. Our company has recently hired a young PhD to do materials research in another department . He is pushing very hard to be allowed to use our SEM at night and on weekends. He has 'used an SEM before' and doesn't see a problem in using ours. It has always been our practice to encourage researchers to come into the lab while their samples are being analyzed, but not to allow individuals to use the instrument themselves. I believe that the analysis is done more effectively by individuals whose expertise is in that area. However, management appears to be leaning towards changing the rules for this individual, and having me train him on the instrument.
We have a four year old JEOL 6400 instrument with a Link eXL analyzer, UTW detector, complete stage automation. We also have a computer system attached for collecting digital images, we do not use polaroid film. While this individual has used and SEM he has not used either a JEOL or LINK system before. I feel that training would be extensive. This is currently the only SEM we have. It is used approximately 12 hours per day by myself and my staff. Quite often EDS analysis is carried out using computer control and stage automation overnight. However, management feels that if someone is willing to utilize the instrument in those few hours a week when it is not being used, they need to take advantage of that to receive a maximum benefit from the asset.
Several years ago we had two older SEMS and due to 'corporate downsizing', we had only one operator. At them time I trained approximately 2 dozen engineers who desired to use the 'extra' SEM on their own. We disposed of the instrument about a year ago when it had not been touched for almost a year. The time I spent in training all of those individuals was probably greater than the time they as a group spent using the instrument. Hindsight being perfect I wish we had kept the old instrument!
Anyway, I would appreciate any input both pro and con from the list regarding opening up an instrument to multiple infrequent users.
For your information, the following two companies also sell the “wedge-polishing” tool, supplies, and various consumables (diamond lapping films/suspensions, mounting wax/glues, etc.) for TEM specimen preparation of materials:
UltraMetrix 4802 E. Ray Rd., No. 23-267 Phoenix, AZ 85044 (602) 706-5745 FAX (602) 496-6505 e-Mail ULTMTRX-at-aol.com
Allied High-Tech Products P.O. Box 4608 2376 E. Pacifica Place Rancho Dominguez, CA 90220 (310) 635-2466 FAX (310) 762-6808
Rene E. Nicholas Spec. Prep. Workshop Coordinator AMC Group
At 1:09 PM 2/9/96 -0500, Bonnie Davis - Kennametal Inc. wrote: } I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM. My situation is that I am part of a materials } analysis group that provides analytical services to corporate } technology. Our group consists of a number of engineers and technicians } with expertise in a variety of analytical areas. Our company has } recently hired a young PhD to do materials research in another department . } He is pushing very hard to be allowed to use our SEM at night and on weekends. } He has 'used } an SEM before' and doesn't see a problem in using ours. It has always } been our practice to encourage researchers to come into the lab while } their samples are being analyzed, but not to allow individuals to use the } instrument themselves. I believe that the analysis is done more effectively } by individuals whose expertise is in that area. However, management } appears to be leaning towards changing the rules for this individual, and } having me train him on the instrument. } } We have a four year old JEOL 6400 instrument with a Link eXL analyzer, } UTW detector, complete stage automation. We also have a computer system } attached for collecting digital images, we do not use polaroid film. } While this individual has used and SEM he has not used either a JEOL or } LINK system before. I feel that training would be extensive. This is } currently the only SEM we have. It } is used approximately 12 hours per day by myself and my staff. Quite } often EDS analysis is carried out using computer control and stage } automation overnight. However, management feels that if someone is } willing to utilize the instrument in those few hours a week when it is } not being used, they need to take advantage of that to receive a maximum } benefit from the asset. } } Several years ago we had two older SEMS and due to 'corporate } downsizing', we had only one operator. At them time I trained } approximately 2 dozen engineers who desired to use the 'extra' SEM on } their own. We disposed of the instrument about a year ago when it had } not been touched for almost a year. The time I spent in training } all of those individuals was probably greater than the time they as } a group spent using the instrument. Hindsight being perfect I wish we had } kept the old instrument! } } Anyway, I would appreciate any input both pro and con from the list } regarding opening up an instrument to multiple infrequent users.
Bonnie, Since we encourage use of the JEOL TEM and SEM by trained users, we run into this situation often. Currently, we require training on our instruments plus a minimum of 15 hours of daytime use prior to an approval of use during off hours. The principal investigator (of the grad. student or postdoc) is also asked to agree to cover costs should any malfunction necessitate lengthy (costly) alignment procedures. I constructed a form for this purpose and the signed agreement is kept with the log book at the scope. This has given me some control over the situation and those users who agree to our demands are usually competent users. If the SEM uses a LaB6 or is a FE, I would be more cautious, i.e., demand more daytime use and obtain written consent to replace the crystal/filament. One last thing to consider---an estimate of costs incurred by your unit during the required "daytime" use if your unit does not charge other units for instrument use. Regards, Rosemary Walsh
Hello all, I am currently doing TEM of ferroelectric materials (PZT, PST etc.) and seem to be seeing domains or domain walls in thin TEM samples but not in thick ones. I know that TEM of ferroelectrics has been done for some years - does anyone on the listserver know of any references which describe the contrast mechanism which makes them visible in TEM?
Many thanks in advance,
RIchard Beanland, GMMTL Caswell, Towcester, Northants NN12 8EQ, UK Tel. +44 1327 356363 Fax. +44 1327 356775 Email richard.beanland-at-gecm.com
All modern EDS systems include in the quantitative procedures an option "standardless analysis". When standardless analysis is performed, tipically the analysis of stainless steel is demonstrated with measuring transition elements Fe, Cr, Ni, Mn, Co and V. The K-lines of these elements span in the range 5 - 7.5 keV where the spectrometer efficiency is constant and close to unity. The product is well behaved, and a reasonably accurate analysis can be obtained, especially if the procedure has been "optimized" for steel analyses. For testing standardless programs we need suitable data base of measured intensity data with detailed information about analysis (accelerating voltage, geometry) and detector constants. Unfortunately no systematic collection of such data to be available in the area standardless analysis. This is a sad situation in comparison with , for example analysis with standards were there is an large collection of data. For this reason we are in our laboratory decided to collect the intensity data for testing standardless programs. Your help in adding new data sets would be much appreciated. The rules for the data in the collection are very simple:
a) only experimental data from certified samples are included.
b) all data must be collected under known experimental conditions: acclerating voltage, geometry (tilt angle, take-off angle) and detector constants (Be window or other thickness, Au layer thickness, Si dead layer thickness and Si crystal thickness).
c) only pure intensity data (after background substraction and peak deconvolution procedure) are included.
d) weight fraction of analyzed elements must be known.
Final database will be available at EMMPDL (Electron Microscopy and Microanalysis Public Domain Library) on anonymous FTP server WWW.AMC.ANL.GOV (Argonne National Laboratory, IL, USA). We have no any commercial interest with this database.
Thank you.
Henrik Kaker Metal d.o.o. SEM/EDS Laboratory Koroska c.14 62390 Ravne Slovenia
Hello all, I am currently doing TEM of ferroelectric materials (PZT, PST etc.) and seem to be seeing domains or domain walls in thin TEM samples but not in thick ones. I know that TEM of ferroelectrics has been done for some years - does anyone on the listserver know of any references which describe the contrast mechanism which makes them visible in TEM?
Many thanks in advance,
RIchard Beanland, GMMTL Caswell, Towcester, Northants NN12 8EQ, UK Tel. +44 1327 356363 Fax. +44 1327 356775 Email richard.beanland-at-gecm.com
IS the Gatan Ion mill still available? Joe Geller 508 887-7000
On Thu, 11 Jan 1996, Gordon L. Nord Jr. wrote:
} Dear Microscopists, } } Because of the recent downsizing of the US Geological Survey the TEM lab } must go. In addition, because the last remanents of the Bureau of Mines (may } it rest in peace) are coming to the USGS in Reston the lab space is needed for } other purposes by next week (gasp). All or part of the following are available } for transfer to US academic institutions. The USGS has not mentioned any } possibility of help with respect to shipping but I could be surprised. } } (1) JEOL 200B Transmission Electron Microscope in working condition (1974) with } STEM attachment (1978). } } (2) Multiple stages including Be double-tilt stage by Gatan and heating stage } by JEOL. } } (3) Technics Ion Mill, 1974 (works) } } (4) Gatan Duo Ion Mill with low voltage guns and cooling stage, 1983 (works) } } (5) Tracor Northern 2000 Multichannel analyser and detector 1978 (works) } } (6) LogEtronics enlarger 1974 (works) } } Contact me at the following address. } } Cheers, } Gordon } } } Gordon L. Nord Jr. } 959 National Center } U. S. Geological Survey } Reston, VA 22092 } } Office: 703-648-6745 } FAX: 703-648-6789 } } gnord-at-mactem.er.usgs.gov } }
Dear Microscopists Could anyone advise me on current developments in the embedding (wax or otherwise) sectioning and staining of botanical specimens for histological and anatomical examination. We have got as far as Histoclear, but most other techniques seem to be more than fifty years old [and working satisfactorily, I may add]. Some of the old stains are fading though and we have to replace old slides, and this might be an opportunity to apply any up to date technique.
Cheers
Stephan Helfer Royal Botanic Garden Edinburgh Scotland UK
I must echo Bob Craig's observations about "external" users. He could have easily written that word-for-word as a fellow employee here. Our back-up plan for extended use of high-demand instruments is to go to an informal swing schedule. Fortunately we have not had to resort to that officially, but most of our gang have a lot of "after hours" time logged. One method we have utilized for increasing throughput on our "automated" instruments is to dial in from home with Timbuktu and AppleTalk Remote Access to check on progress. If something has gone amuck, we can either fix it remotely or zip in to work, fix it and resume the experiment. It's not pleasant to come in at midnight instead of going to bed, but our customer's anxiety (hence ours!) is greatly diminished the next day.
Bill ========================== Bill Heeschen/Analyt. Sci.- Mat. Char. ----- 1897-F Building / The Dow Chemical Co. --- --- Midland, MI 48667 U.S.A. --- D O W --- phone: (517)636-4005 fax: (517)636-5453 --- --- Email: waheeschen-at-dow.com ----- ========================== R
Subject: Time:11:04 AM OFFICE MEMO Antibody to TRH Date:2/12/96
Hello Everybody, I am looking for antibody to Thyrotropin-releasing-hormone for immunocytochemistry which will work on rat brain. Does anyone know where I can get it? I. Polyakov NASA-Ames Research Center FAX: (415)604-0046 Email:Igor_Polyakov-at-qmgate.arc.nasa.gov
To: Microscopy ListSer {Microscopy-at-MSA.Microscopy.Com}
This has been a problem for us at times. The real issues are; the ability of people to work together, and the cost of maintenance, and keeping the instrument in top condition. We find it possible to have up to about 3 operators, as long as they can work together well. They also need to be in the same department (i.e. financed together). Pride in the condition of the instrument and the quality of results is critical.
There can be other issues, like job security. If allowing someone access you are putting someone else out of work...........
When the machine breaks, whoever had their hand on it, must immediately get it fixed. This works best when there's a service contract.If there's an assigned technician, this may not be a problem for you.
My overall impression is that the number of operators must be limited to a couple who can work together well, and not quibble over who did what.
{ } { { } } ===========} Dave King { { { } } } ------------------------------------------------------ { { { {
We have a researcher who wishes to determine if coal fly ash is present in river water and river sediment. By LM I detect some "pearly" particulates not present in control river sediment. Next step will be examination by SEM.
My question: is there an established protocol for detecting and quantifying fly ash in river water and sediment?
I will post any responses not sent to the server directly - unless otherwise directed. Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Message-ID: {199602122331.SAA16597-at-IndyNet.indy.net} To: Richard Beanland +44 1327 356363 {richard.beanland-at-gecm.com} , Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
[various snips] } I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM. } Our company has } recently hired a young PhD to do materials research in another department . } He is pushing very hard to be allowed to use our SEM at night and on weekends.
He has to be trained very well to use the machine when no staff mem- ber is around. It is much easier to have him use the SEM during the day, and I would insist that he do so after training until you are absolutely satis- fied that he is competant before allowing him to work without a net.
} He has 'used } an SEM before' and doesn't see a problem in using ours. It has always } been our practice to encourage researchers to come into the lab while } their samples are being analyzed, but not to allow individuals to use the } instrument themselves.
For our facility--a NIH biotech resource--we have several clas- sifications for outside users, from novice, for whom the staff does every- thing, to expert, who can use the instrument without supervision. We train the user in many steps, the first being the basics for tilting and transla- ting the specimen, focussing and taking a picture, then changing specimens, later changing film, starting and shutting down the scope, etc. All this takes several months for an in-house user, and longer for someone who visits occasionally.
} This is currently the only SEM we have. It } is used approximately 12 hours per day by myself and my staff. Quite } often EDS analysis is carried out using computer control and stage } automation overnight. However, management feels that if someone is } willing to utilize the instrument in those few hours a week when it is } not being used, they need to take advantage of that to receive a maximum } benefit from the asset.
There is a large negative benefit if anything goes wrong; try to impress management that a conservative approach is best.
} Anyway, I would appreciate any input both pro and con from the list } regarding opening up an instrument to multiple infrequent users.
This will take a *lot* of your time. Good luck, you'll need it. Yours, Bill Tivol
Yes, there are various programs on the market. Depending on your budget, and your need for continued support. Probably the most ecconomic of the bunch is Image Pro Plus, soon to be available using 32 bit, and windows 95.
The second PC based system, that I find very intuitive and has excellent support is Kontron's KS 400 IA system. Kontron KS versions are distributed and supported by Zeiss, Thornwood NY. The best person to contact is Uli Kolhaaus, or Elise Shumpsy at Zeiss.
Gregory Argentieri Sandoz Pharmaceuticals Corp East Hanover, NJ 201-503-8617
Hello everyone, I have been embedding pinon pine root tips for mycorrhiza research. We are close to solving most of our problems but still seem to have some left. And I was wondering if some fresh ideas may help. Problem: Portions (it seemed to be mainly the cortical tissue) of the root tip will not completely infiltrate and polymerize. The mantel looks okey and central vascular area seems to embed okey. The cortical cells seem to be our main source of infiltration problems. Some of the samples float (even after hours or days in a vacuum). At first, we used Spurrs resin with graded Etoh. We have not used acetone or propylene oxide (yet).
Figuring we were not getting all the water out of the tissue nor getting our resin into the tissue we have tried the following procedures: 1. Cut our root tip sample even smaller. 2. We obtained some Z-6040 which has been mentioned several times on this list for helping to infiltrate impervious biological specimens. We used it at 1% as advised by Virginia Lindley's article. But we also continued its use in the LR White resin as recommended by Stacie Kirsch at EMS. 3. We found we had to extend our Abs. ETOH times to over night and several days of changes of LR White. We switched to LR White as recommended in Lindley's article.
At this point, we had some success, but still had some root tips that wanted to float (even under vacuum). I have not worked with root tips or mycorrhiza before, but I may be doing more in the future. Do we need to use acetone or propylene oxide to remove all the water?
Thank-you for any comments or helpful suggestions. If you should like respond to me at my local address, I shall summarize for the listserver.
Good day, Marilee
Marilee Sellers Manager, Electron Microscope Facility Northern Arizona University Flagstaff, AZ 86011 E-mail marilee.sellers-at-nau.edu
I am searching for the most comprehensive articles about critical point drying techniques. All suggestions are appreciated.
Any problems with going directly from ethanol into carbon dioxide or is amyl acetate useful in between? My specimens look OK using just ethanol.
Also: I am experiencing a run of failing (most often between 400 and 800 psi) front window Dowty seals on my drying unit even if I do only 'dry' (carbon dioxide only) test runs. Is anyone else having this problem? This device has been operating for 17 years with few leaks. All sealing surfaces are smooth and clean.
Thank you in advance.
Jim Romanow Electron Microscopy Facility Physiology and Neurobiogy Department The University Of Connecticut Storrs bsgphy3-at-uconnvm.uconn.edu
Message-Id: {199602122054.OAA28915-at-watson.bcm.tmc.edu} X-Sender: joiner-at-bcm.tmc.edu X-Mailer: Windows Eudora Version 2.1.1 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Thanks, Gib, for your message.
"....that some stray electrostatic charges floating around on these low humidity winter days can precipitate beautiful hex lead crystals just doesn't click for me...."
As I had said before, I also don't think that static charges would cause inappropriate stain precipitation, but then as you can see from the response on the listserver, a lot of people have experienced this problem and the cause(es) have not been determined. At this point I am willing to try anything and the suggestion didn't seem unreasonable. ***********************
"....I say chuck the whole damn works and switch to the Ted Pella "Grid Stick" kit to do your uranium and lead staining...."
We have tried Pella's Grid Stick. However that procedure washes the sections off of our grids. Our type of work necessitates the use of large sections and 200 mesh "Super Grids" (grids with very thin grid bars). The use of these grids results in much reduced area of contact between grid and section, and the sections just do not adhere to the grids well. Now, if you have any suggestions as to how we can get the sections to stick to the grids better, I would appreciate them very much. Then perhaps we could institute a more vigerous wash. ***********************
"....For lead, I use the Sato triple lead stain, not straight Reynold's lead citrate...."
I do know of the Sato tripple lead stain, but would appreciate the reference, if you have it handy. We have tried both the Reynold's recipe as well as the recipe from Dr. Chandler Fulton. I don't have that reference. However nothing could be simpler. To 10 ml of decarbonated, distilled water you just add 0.02 gm lead citrate powder (EMS cat no. 17800), followed by 0.1 ml of 10N NaOH. This stain worked very well for me throughout my graduate school experience.
Joiner Cartwright, Jr., Ph.D.
Director, Electron Microscopy tel.: (713)798-4658 Department of Pathology, Rm.286-A FAX: (713)798-3945 Baylor College of Medicine Internet: joiner-at-bcm.tmc.edu One Baylor Plaza Compuserve: 71555,1206 Houston, Texas 77030 U.S.A.
I'd like some input, opinions etc. on the pitfalls of having multiple operators on an SEM. My situation is that I am part of a materials analysis group that provides analytical services to corporate technology. Our group consists of a number of engineers and technicians with expertise in a variety of analytical areas. Our company has recently hired a young PhD to do materials research in another department . He is pushing very hard to be allowed to use our SEM at night and on weekends. He has 'used an SEM before' and doesn't see a problem in using ours. It has always been our practice to encourage researchers to come into the lab while their samples are being analyzed, but not to allow individuals to use the instrument themselves. I believe that the analysis is done more effectively by individuals whose expertise is in that area. However, management appears to be leaning towards changing the rules for this individual, and having me train him on the instrument.
We have a four year old JEOL 6400 instrument with a Link eXL analyzer, UTW detector, complete stage automation. We also have a computer system attached for collecting digital images, we do not use polaroid film. While this individual has used and SEM he has not used either a JEOL or LINK system before. I feel that training would be extensive. This is currently the only SEM we have. It is used approximately 12 hours per day by myself and my staff. Quite often EDS analysis is carried out using computer control and stage automation overnight. However, management feels that if someone is willing to utilize the instrument in those few hours a week when it is not being used, they need to take advantage of that to receive a maximum benefit from the asset.
Several years ago we had two older SEMS and due to 'corporate downsizing', we had only one operator. At them time I trained approximately 2 dozen engineers who desired to use the 'extra' SEM on their own. We disposed of the instrument about a year ago when it had not been touched for almost a year. The time I spent in training all of those individuals was probably greater than the time they as a group spent using the instrument. Hindsight being perfect I wish we had kept the old instrument!
Anyway, I would appreciate any input both pro and con from the list regarding opening up an instrument to multiple infrequent users.
In a commercial organization, an instrument is a resource for all. The question can not be is a person allowed to use a resource because they are or are not a member of a particular work group. The question must be how do we maximize productivity. Working within this framework, here are two observations to balance:
It is typically counter-productive to have relatively inexperienced users operate instruments which are either finicky or complex.
It is typically counter-productive to have expert users repeatedly perform a relatively simple analysis.
Now for the reduction to practice. We have trained researchers from a variety of groups to use our microscopes, OM, SEM, and TEM. This has included training an inexperienced researcher on a FEG-SEM. We have never had a mishap which cost us more than a few hours of instrument time. The OM and SEM training has benefited productivity. However, TEM training has been another story. Inexperienced researchers were repeatedly retrained, resulting in an inefficient use of expert personnel.
Good luck to all,
Craig Garrison The Dow Chemical Company Bldg. B-1225 2301 N. Brazosport Blvd. Freeport, TX 77541
I am looking for a postdoctoral researcher to fill the following post. Please could any interested parties contact me directly.
Thankyou, Amanda Petford-Long.
************************************************************* Advertisement for postdoctoral research post in the Department of Materials, University of Oxford, UK funded by Hewlett-Packard.
A post is available, in the first instance for 1 year, to study layered films for magnetoresistive read head applications. This will form part of an ongoing collaboration with Hewlett-Packard Laboratories in Palo Alto, involving both experimentalists and modellers within the Department of Materials.
The research will involve the characterisation of the microstructure and magnetic domain structure, by electron optical techniques, of films grown at HPL by sputter deposition. The films under investigation will be spin-valve structures, and the interest lies in the role of microstructure on giant magnetoresistance and magnetic properties. The microstructure of the films will be analysed using high resolution electron microscopy, and we have already had some considerable success in quantifying the nature of the interfaces between the layers in the spin valve using imaging processing techniques on HREM images, and would hope that this work could be continued. The magnetic domain structure of the films will be studied using our extensive Lorentz transmission electron microscopy facilities. These include facilities for in situ magnetising and heating, and also for the collection of quantitat- ive magnetisation maps. Some experience in electron microscopy (preferably HREM) is therefore essential.
The post is fully funded by HPL on a rolling grant basis, so that further funding after the 1 year period is extremely likely but cannot be guaranteed at this time.
The post is available from 1st July 1996 and will be on the University salary scale GBP14,317 - 21,511 p.a.
Interested applicants should submit their applications, including a curriculum vitae and the names and addresses of two referees to the Administrator, Department of Materials, Parks Road, Oxford OX1 3PH, UK. Please quote ref. AKPL. The University is an Equal Opportunities employer.
For informal queries please contact Dr Amanda Petford-Long in the Department of Materials. e-mail address: amanda.petford-long-at-materials.ox.ac.uk
Others you might try:Arnel Products Company (Tel. 212-620-4622), Biogenesis Ltd. (Tel. 603-887-4600, Email biogenesis-at-ltd.co.uk), Chemicon International (Tel. 800-437-7500), Eurodiagnostics Lab (Sweden, fax 46-409231000), Cappel-Organon Technica (Tel. 800-523-7620) and UCB Bioproducts (carried by Accurate Chemical and Scientific Corp, see above).
All these were raised in rabbit, so they should work on rat brain, although I havn't tried any.
This is from the 1996 Lindscott's Directory of Immunological and Biological reagents (Order from Lindscott's Directory, 4877 Grange Road, Santa Rosa, CA 95404; phone (707) 544-9555, Fax (415) 389-6025), one of the best places to start looking for this kind of information.
Richard D. Powell ************************************************************* NANOPROBES, Incorporated 25 East Loop Road, Suite 124, Stony Brook, NY 11790-3350, USA http://www.lihti.org/research/ecodev/incubten/nano/home.html
Message-Id: {199602131409.KAA17548-at-Snoopy.UCIS.Dal.Ca} Comments: Authenticated sender is {RMACKAY-at-ac.dal.ca}
RE: Multiple Users
Dear Bonnie,
We have operated a JEOL 733 electron microprobe ( presently with a Link eXL system ) for the past 11 years, and prior to that a Mark V. Our policy has always been to allow multiple users including graduate and undergraduate students. In 25 years we have had only one bad experience.
Best Regards,
Bob MacKay Robert MacKay Department of Earth Sciences Dalhousie University Halifax, Nova Scotia, Canada B3H 3J5 Tel: 902 494-7087 e-mail rmackay-at-ac.dal.ca
} Marilee, A microwave protocol is especially well suited for your work. I } recommend checking the recent article by Giberson RT, Demaree RS, Jr. Microwave } fixation: understanding the variables to achieve rapid reproducible results. } Microsc Res Tech 1995;32: 246-54. } } Gary R. Login, Beth Israel Hospital, Pathology } 617-667-2034; glogin-at-bih.harvard.edu
Gary, This is one alternative I hadn't even considered. Thank-you. I do not have a research microwave. But will see if ours would do. Marilee
Marilee Sellers Manager, Electron Microscope Facility Northern Arizona University Flagstaff, AZ 86011 E-mail marilee.sellers-at-nau.edu
I have a question which maybe someone can answer. Assuming constant angle during ion-beam thinning, which of these two leads to less nett damage (point defects etc) within a sample, assuming that both remove the same amount of material: a) A longer time at low energy b) A shorter time at higher energy ?
You can also try Visilog from Noesis Vision Inc. It provides the widest selection of algorithms on the market along with an easy to use GUI and macro language. We are soon coming out with a 32 bit version on both Unix and Windows NT. You can contact me for more information. Luc Nocente Tel: 514 345 1400 Noesis Vision Inc. Fax: 514 345 1575 e-mail: ln-at-noesisvision.com http://www.cam.org/~noesis 6800 Cote de Liesse, Suite 200 St-Laurent, PQ H4T 2A7,Canada
} Marilee, } } A year or so ago, we were working on a similar sounding project looking } at mycorrhizal associations in pine roots (different pine). We were } interested mainly in the composition of some inclusions but prepared material } for general morphology too. We used a "standard glut/Fa" primary fix and an Os } tet secondary followed by acetone dehydration and embedment in Spurr'Us or } our own variant thereof (VCD and Quetol). We had no problem with infiltration } or sectioning. } } Here's my twenty questions: } } 1. Do you use vacuum during fixation? I generally bounce the tissue } during primary fixation by cycling the vacuum until the tissue stays } submerged up to the boiling pressure of the fixative.
We have not used vacuum during fixation. Most of the time the tissue did not float until after the Abs. Etoh. We shall include this.
} 2. Did you try to embed any of the floating material? I generally don't } try to embed "floaters". My thinking is that if they have enough air in them } to float after Os tet density increase, they've got too much air for good } transfer of any thing into or out of the tissue.
We did embed some floaters. And they were poorly infiltrated.
} 3. When do you notice the problem in infiltration? i.e. during } sectioning, or after observation. I've had occasional problems with LR white } bonding to cell walls. Usually we can get enough to hold still for a picture } if we carbon coat them after labeling and staining and before viewing. The } stuff is supposed to be able to handle some water as an impurity, but I'm not } sure I believe it. } 4. What kind of knife do you use? I used glass since our root samples came } from the real world and had entrained some sand as well as air. If yours don't } have anything hard in them (or you'Uve got lots of money;-)) might use a } diamond if youUre ot now. I cut a lot of hydroponically grown roots for class } and generally get O.K. sections in regular resin with my diamond knife. Even } they hydroponically grown tips have considerable exhaustible air in them.
We notice the problems immediately upon sectioning. These root tips are from sandy soil, and the tips are cleaned as much as possible before processing. As a precaution, we use glass knives to cut them.
} John Heckman } TEM supervisor } Center for Electron Optics } Michigan State UniversityMarilee, } } heckman-at-pilot.msu.edu
Thank-you John. We will be incorporating your suggestions. Marilee Marilee Sellers Manager, Electron Microscope Facility Northern Arizona University Flagstaff, AZ 86011 E-mail marilee.sellers-at-nau.edu
View From A University Central Service Microscopy Lab
As the Director (and only employee) in a lab with a TEM, SEM, confocal, imaging equipment, photographic darkroom, and the usual ancillary equipment, and no service contracts, I could not function if I had to do everything for users. Ninety percent plus of my users operate the equipment themselves. However, less than 5 users are 100% qualified to operate the EM's when I am not present. I insert and remove specimens, and for the TEM, process the negatives. Obviously, users undergo training, and during training one can pick up cues as to the user's future behavior. Our equipment is very user friendly, and that makes a big difference. Obviously in industry where down time is unacceptable, this approach would not work. Bruce Cutler, Microscopy Lab, Univ. Kansas, Lawrence
We have a couple of users who want to look at calcium and/or pH changes on suspension cells (culture cells). Does anyone know of specific tricks for trying to stabilize single/suspension cells in a perfusion chamber. We would like to follow a single/small group of cells throughout treatment with different agents. We have tried both charged and coated coverslips (silane and gelatin) without sucess. Any and all help would be greatly appreciated.
TIA,
C. Michael Stanley, Ph.D. Coordinator, Associate Director Molecular Cytology Core Facility Molecular Biology Program 2 Tucker Hall University of Missouri-Columbia Columbia, MO. 65211 (314) 882-4895 fax= 314-882-0123
} The silicon 'dead layer' in a detector is a mathematical concept to } describe a region in the detector of reduced sensitivity to the incoming } x-rays (the Glasgow people coined a term for this which now eludes me). } The 'dead layer' varies with the detector bias, and cannot be measured } directly - it is possible only to deduce it from experiments fitting } measured bremsstrahlung shapes to theoretical predictions.
Dear Tony, The dead layer of a Si detector can, indeed, be measured experi- mentally--at least in principle. Using an alpha source, measure the energies when the alphas are incident on the detector both normal to its face and at another angle for which the alphas still penetrate the dead layer. From these measurements the thickness of the dead layer (plus the gold) can be determined--at a number of bias voltages if desired. Personally, I'd never subject my detector to this measure- ment, but it is possible. We used to do this all the time for detec- tors when I was in grad school. Yours, Bill Tivol
This is an interesting problem. I have had to train other users to use my SEM, but have always found it to be a waste, since they didn't use it enough to remember the training. I had to go over things each time they wanted to use it.
However, if you are forced to let this person use the SEM, here are some guidelines. Mark the saturation point clearly, and make it a rule to not go past that mark. Have a log book handy for the part time user to record any 'unusual' occurences in operation, or to write questions down. Have an emergency number to call for help. (I know this is a pain for you, but could save on instrument downtime and repair costs.)
That's all I could think of right now, but I hope it helps.
Regards, Melanie Behrens Senior Chemist Texaco, Inc. Beacon, NY behrema -at- Texaco.com
Hello all, I need to obtain the address and fax number of 'Triarch Slides' in Wisconsin, USA if they are still in business. Any help would be appreciated. Regards,
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Dear All,
Many thanks to all those who responded to my query re mode of action of toluidine blue in quenching autofluorescence, particularly Bill Tivol and Bridget Southwell who gave an answer that I can trot out if ever asked again! In response to those who requested further details of the way I use tol. blue: it is applied as a 0.01% (w/v) solution made up in phosphate-buffered saline (pH nominally 7.3 - 7.6) to the sections (I routinely use 6 um thick methacrylate-embedded Aesculus hippocastanum root sections). Leave the stain to impart a 'light-blue' colour to the sections, ie, let them 'just stain', that takes about 90 sec with my tissue. Sections then mounted in anti-fade mountant and viewed - stored if necessary in a fridge (at c. 4 C). [Sections having been processed for visualisation of tubulin using FITC-linked secondary antibody, and usually stained with DAPI prior to tol. blue-quenching] "It works for me"... I do not know if tol. blue itself fluoresces under some excitation wavelengths, but it does reduce some of the background/native autofluorescence present in my tissue (for Sally Stowe). The reference I have used for this procedure is Baluska, F, Parker, JS, Barlow, PW, 1992. Specific patterns of cortical and endoplasmic microtubules associated with cell growth and tissue differentiation in roots of maize (Zea mays L.). J. Cell Sci. 103, 191-200. Of course, if anybody can provide an earlier (more definitive?) reference to its first use, I would be grateful. Hope the above is useful,
I'm posting this request for a friend that needs to buy a light microscope. He specifically is looking for a microscope with a large stage that will allow a long working distance for using a micromanipulator. Does anyone have a used microscope that they want to sell? Also, does anyone know the address or telephone number of equipment brokers that might have light microscopes? If so, please respond to Irv Widders, Michigan State University, E-mail 22457iew-at-msu.edu
I have found that (a), a longer time at low energy, is better for my samples- III-V semiconductors.
Ciara
On Tue, 13 Feb 1996, L.D.Marks wrote:
} I have a question which maybe someone can answer. Assuming } constant angle during ion-beam thinning, which of these two leads to } less nett damage (point defects etc) within a sample, assuming that } both remove the same amount of material: } a) A longer time at low energy } b) A shorter time at higher energy } ? }
Hi Bonnie, One point that everyone seems to have missed on this subject of multiple users is output quality. There is no substitute for the skill gained with experience. We tried running two SEM's in our lab, and found the overall quality of the images produced by our "trained" multiple users was not as good as the images produced by an experienced technician.
Cheers :) George Department of Earth and Atmospheric Sciences University of Alberta Edmonton, Alberta T6G 2E3 Canada ph: 403-492-5746 fax: 403-492-2030
There has been alot of comments on the matter of multi-user of and SEM. I have been on both sides of that fence. I operated and SEM for years at the Bureau of Mines until I took a research position. With my position I had the need for SEM work. This I was able to do most by myself. I am now with a company that does not have an SEM so I have to use the one at the university. While I was the operator I was very choosey about who could use it. Only a hand full of people asked to use it and when I started the training they decided it was to much to remember and that I should do it because they did not have the time to do it correctly. Now that I use someoneelses I am very carful to follow all the instructions, I want to be able to come back. I also felt very uncomfortable the first few times I was left alone with this instrument, now I am familiar with it and use it much more. I guess what I am trying to say is some people have no idea about an instrument and are hot shot scientist, newly graduated PhD's and others. These are people to be aware of. Multi-users are okay depending on the type of business you are in and the personnal. I thought an opion from the other side should be expressesd.
Clarissa Vierrether Analytical Development Chemist The Doe Run Company P.O. Box 500 Viburnum, MO 65566 1-573-244-8109
As Tony Garratt-Reed pointed out the 'dead layer' in an EDS detector is an unfortunately chosen name. As first shown by Goulding (Nucl.Instrum. and Methods 142, 213, 1977) the 'dead layer' is simply the region over which drift anf diffusion approximately cancel each other. The behavior of this region is crucial when good resolution at low energy ( {2keV) is required. The MAS Fiori Memorial volume "Xray Microanalysis in Electron Beam Instruments" published by PLENUM contains two papers which discuss this in detail, and also contain a bibliography of relevant references - one
Joy DC, "Modeling the Energy Dispersive X-ray detector", in "Xray Spectrometry in Electron Beam Instruments", ed D B Williams, J I Goldstein and D E Newbury, (Plenum Press:New York), 53-64, (1995)
which discusses a Monte Carlo simulation of this type of effect, and a second by Jon McCarthy of NORAN which examines the practical implications of 'dead layer' behavior. The dead layer can be estimated in several quite convenient ways, including spectral measurements, matching to simulation ot, best of all, by an electron beam measurement described in
Joy D C, "The EDS Detector - A Quantitative Model", Rev. Sci. Instrum. 56, p1772-1779, 1985
As an interesting aside note that the dead layer in silicon at room temperature is only 700 angstroms or so compared to 2800 A or more at liquid nitrogen temperatures
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Bonnie,
The Integrated Microscopy Resource (IMR) is an NIH biomedical research technology resource. We have a Hitachi S-900 FESEM with cryo-capability and a YAG backscatter detector. As one of the few microscopy facilities open for biologists, the novel instruments at the IMR are made available to users world wide for biomedical research projects. Our user base consists of senior investigators, post-docs, and graduate students.
As an NIH resource, the use of instruments is reserved for medical research projects that require the specialized aspects of the available instruments. Our SEM usage policy consists of three levels of approval: basic use, backscatter detector use, and cryo-stage use. I provide user training and supervise the use of the SEM for the first 20 hours. After this time, approved users may be upgraded to unsupervised users who are permitted to work independently after hours. Facility keys (either short or long term) are available for unsupervised users. For this system to work it is critical to give each user comprehensive instructions on use of the equipment, to establish clear rules with regard to general use of the facility, and most importantly to have an individual with the authority to monitor and enforce these rules.
Our open use policy has been in place for 9 years with the SEM and 25 years at our facility. Project approval and user training prior to equipment usage has resulted in much success and very few problems. When problems have arisen the solution has been for me to re-train the user and continue to supervise them for longer time.
Hope this information answers your question.
Ya Chen
========================================================================= \ / Assistant Researcher, Cryo/SEM Coordinator TEL : 608-263-8481 \ / __ Integrated Microscopy Resource (IMR) FAX : 608-265-4076 \/ / / University of Wisconsin-Madison / / / 1675 Observatory Drive #167 Email:YChen-at-macc.wisc.edu / /__/_ Madison, WI 53706 Email:chen-at-calshp.cals.wisc.edu
IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr/imr.html IMR Symposium on integrated microscopy '96: Sept. 20-22, 1996 =========================================================================
Anyone have any good references re: Calicivirus? Especially interested in classical virilogical studies (morphology, chemical characterization, pathology, vectors, control measures) and current press releases on the "epidemic" in Australia. Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Hello all, Thank you to all who contributed info on Triarch. If anybody knows of other competitively priced prepared slide manufacturers I would appreciate their contact details also. Regards,
Hi Everyone, I have a JEOL 5300 SEM to which a beam controlled image acquistion system has been added. Images are acquired at 640x512 by frame averaging. It takes 1min 20sec. to collect five frames. This is about the same time it takes to do a Polaroid. Here is the problem. At magnifications above 5000x the images are fuzzy due to some kind of drift. At lower mags the drift is not noticeable. Polaroids do not show the drift. The drift rate is 100-500u per five minutes. I have the SEM manufarturer working on this too. So far I have not gotten acceptable results Now for the questions. Has anyone had this problem on other image acquisition systems? Am I trying to do something that can't be done due to drift that all SEMs have to some degree? Thanks for your answers in advance.
Gregory Rudomen Greg-at-umic.umic.sunysb.edu University Microscopy Imaging Center S.U.N.Y. at Stony Brook
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I am looking for good references whose discuss about the experimental setup (hardware, specimen preparation, experimental conditions, bias etc.) for voltage contrast and EBIC experiments. I am particularly involved in the fabrication of III-V high-speed devices for signal capture (CCD and Sample&Hold).
Mario
******************************************************* Mario Caron, M.Sc.A. ing. Research assistant Engineering Physics dept Laboratory for the Integration of Sensors and Actuators Ecole Polytechnique de Montreal C.P. 6079, succ. `centre-ville` Montreal, Quebec H3C 3A7 http://lisa.polymtl.ca
".....to form yellow, iridescent hexagonal prisms of lead iodide....."
I stained one grid with our lead stain alone, and another with our uranyl acetate stain alone. The lead stained grid had the hexagonal deposits and the other did not. So it's not the uranyl acetate. It's interesting that lead iodide forms hexagonal crystals. *******************
".....Perhaps these are your crystals - a bit of a long shot since you did not mention iodine in our elemental analysis of the crystals...."
No, there is no iodine in either of our stains, or any of the sample preparation steps; and I did not get an iodine peak with the microprobe. *******************
I have borrowed an anti static gun like the Zerostatic Eliminator that you suggest. We are in the process of trying it out. At least the grids don't jump up and cling to the lid of the plastic petri dish any more.
I'd like some input, opinions etc. on the pitfalls of having multiple operators on an SEM. My situation is that I am part of a materials analysis group that provides analytical services to corporate technology. Our group consists of a number of engineers and technicians with expertise in a variety of analytical areas. Our company has recently hired a young PhD to do materials research in another department . He is pushing very hard to be allowed to use our SEM at night and on weekends. He has 'used an SEM before' and doesn't see a problem in using ours. It has always been our practice to encourage researchers to come into the lab while their samples are being analyzed, but not to allow individuals to use the instrument themselves. I believe that the analysis is done more effectively by individuals whose expertise is in that area. However, management appears to be leaning towards changing the rules for this individual, and having me train him on the instrument.
We have a four year old JEOL 6400 instrument with a Link eXL analyzer, UTW detector, complete stage automation. We also have a computer system attached for collecting digital images, we do not use polaroid film. While this individual has used and SEM he has not used either a JEOL or LINK system before. I feel that training would be extensive. This is currently the only SEM we have. It is used approximately 12 hours per day by myself and my staff. Quite often EDS analysis is carried out using computer control and stage automation overnight. However, management feels that if someone is willing to utilize the instrument in those few hours a week when it is not being used, they need to take advantage of that to receive a maximum benefit from the asset.
Several years ago we had two older SEMS and due to 'corporate downsizing', we had only one operator. At them time I trained approximately 2 dozen engineers who desired to use the 'extra' SEM on their own. We disposed of the instrument about a year ago when it had not been touched for almost a year. The time I spent in training all of those individuals was probably greater than the time they as a group spent using the instrument. Hindsight being perfect I wish we had kept the old instrument!
Anyway, I would appreciate any input both pro and con from the list regarding opening up an instrument to multiple infrequent users.
These things help a great deal when using a glass knife. We use lecithin in the epoxy accelerator . ( use eual weight of lecithin and accelerator, then double the volume for use) A glass knife will last a long time if you have it in the resin. It was a great help to students just learning to section, They could get usable stuff the first day at the microtome. It can give a blotchy look to empty resin areas or those that have little density, but most tissues are "busy " enough that you never notice it. See Mollenhauer , 1986, J. EM Tech 3:217-222 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Dear Bonnie, I run a Materials Engineering EM lab at the University of British Columbia, with two SEMs and a TEM. Most of the researchers who use the instruments are graduate students and Research Engineers. I have always encouraged these people to do their own research on the SEMs, as much as possible, since only they know exactly what they want and that way one me can keep three instruments optimally used. After I show them how to use the SEM, I watch and encourage them to ask me for further instruction for picture taking, higher mag, etc. Some people, who have a lot of work to do, may gain my permission to use the SEM after hours, but only after I feel they have gained a lot of experience during working hours, when I am there to supervise, and only after I have instructed them on how to properly shut down and what to do in the event of problems. I also purchased the instruments originally with ease-of-use in mind. They are fully automatic and easy to get good results on. I must feel that the person has a good understanding of the important issues and knows how to properly handle the instrument. Mind you, I don't have a field emission SEM. I have had damage, but only once in 15 years and it could have happened in normal hours.. It also helps to scare them, be "the ogre of the EM lab". Mind you, we are a teaching lab, so the learning is as important as the doing. I know many EM operators do not like the idea of letting any ham-fisted graduate student or engineer loose on their instrument, but modern SEMs really can be operated by any intelligent being. So long as you satisfy yourself as to this person's knowledge, experience and caring, I'd see no harm. Hope this helps.
Regards, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
Hello Everyone, I'd just like to put in my two cents on the subject of multiple users of an SEM (or in my case an electron microprobe)
Our instrument is used at least 16 hours a day and since I'm only paid for eight almost all our users are trained to use the machine independantly. This allows them faster access to the machine and saves them or their advisors money. The only people we don't encourage to become independant users are those who will only use the machine once or twice. Most users require 3-4 daytime shifts before i will let them use the machine independantly (I give them my home phone number too).
In the six years I've been running the lab I can't recall one incident where the machine has actually been damaged by inexperienced users. On several occasions it has been temporarily put out of commission (computer problems, etc) but usually I can get it back on line immediately the next day. Although there is the potential for a user to damage the instrument (i.e during a sample change) most of the rest of the instrument is fairly fool proof. It is easy to screw up your analyses, but difficult to hurt the machine.
I'm sure the machine would last longer and require less service if I was the only one operating it, but we don't have that option. I also feel that users are much better off acquiring their own data, they know what they want and can change their strategy if they find something unexpected
I have prototype LVSEM that employs the stage mechanism from a Philips EM 430 TEM. I would like to purchase a cold stage for this instrument if I can find one (standard Philips rod). Anyone out there about to "decommission" one?
Jim Pawley
**************************************** Prof. James B. Pawley, Ph. 608-263-3147 Room 1235, Engineering Research Building, FAX 608-265-5315 1500 Johnson Dr., Madison, WI, 53706 JBPAWLEY-at-FACSTAFF.WISC.EDU
At 1:09 PM 2/9/96 -0500, Bonnie Davis - Kennametal Inc. wrote: } I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM. My situation is that I am part of a materials } analysis group that provides analytical services to corporate } technology. Our group consists of a number of engineers and technicians } with expertise in a variety of analytical areas. Our company has } recently hired a young PhD to do materials research in another department . } He is pushing very hard to be allowed to use our SEM at night and on weekends. } He has 'used } an SEM before' and doesn't see a problem in using ours. It has always } been our practice to encourage researchers to come into the lab while } their samples are being analyzed, but not to allow individuals to use the } instrument themselves. I believe that the analysis is done more effectively } by individuals whose expertise is in that area. However, management } appears to be leaning towards changing the rules for this individual, and } having me train him on the instrument. } } We have a four year old JEOL 6400 instrument with a Link eXL analyzer, } UTW detector, complete stage automation. We also have a computer system } attached for collecting digital images, we do not use polaroid film. } While this individual has used and SEM he has not used either a JEOL or } LINK system before. I feel that training would be extensive. This is } currently the only SEM we have. It } is used approximately 12 hours per day by myself and my staff. Quite } often EDS analysis is carried out using computer control and stage } automation overnight. However, management feels that if someone is } willing to utilize the instrument in those few hours a week when it is } not being used, they need to take advantage of that to receive a maximum } benefit from the asset. } } Several years ago we had two older SEMS and due to 'corporate } downsizing', we had only one operator. At them time I trained } approximately 2 dozen engineers who desired to use the 'extra' SEM on } their own. We disposed of the instrument about a year ago when it had } not been touched for almost a year. The time I spent in training } all of those individuals was probably greater than the time they as } a group spent using the instrument. Hindsight being perfect I wish we had } kept the old instrument! } } Anyway, I would appreciate any input both pro and con from the list } regarding opening up an instrument to multiple infrequent users.
Bonnie, Since we encourage use of the JEOL TEM and SEM by trained users, we run into this situation often. Currently, we require training on our instruments plus a minimum of 15 hours of daytime use prior to an approval of use during off hours. The principal investigator (of the grad. student or postdoc) is also asked to agree to cover costs should any malfunction necessitate lengthy (costly) alignment procedures. I constructed a form for this purpose and the signed agreement is kept with the log book at the scope. This has given me some control over the situation and those users who agree to our demands are usually competent users. If the SEM uses a LaB6 or is a FE, I would be more cautious, i.e., demand more daytime use and obtain written consent to replace the crystal/filament. One last thing to consider---an estimate of costs incurred by your unit during the required "daytime" use if your unit does not charge other units for instrument use. Regards, Rosemary Walsh
I tried almost every possibility on our Baltec RES010. I have found that (a), a longer time at low energy with one gun set to a low angle and one gun to a rather high angle worked best on my ceramic materials.
Andreas
______________________________________________________________ Andreas Loewe Tel: +49-228-550-355 University of Bonn Fax: +49-228-678-413 Insitute for Inorganic Chemistry email: loewe-at-uni-bonn.de Inorganic Material Research Roemerstr. 164 53117 Bonn Germany http://www.elmi.uni-bonn.de/ ______________________________________________________________
I'm responding to Greg Rudomen's posting about fuzzy digital images on his JEOL 5300.
Greg, can you give us some more details on your problem? It sounds like you are using a slow scan to do your frame averaging since you say it takes 1 min. 20 sec. to collect five frames.
Are you really seeing drift? Drift will generally be blurred in one direction; if the images are just not sharp, that may be another problem.
When you say the Polaroids do not show the drift, are these Polaroids of the analog or the digitized image? If the analog image is okay, something is happening during digitization.
Another item to check is the power and magnetic field situation in your lab. If the drift is more noticeable at low accelerating voltages, you may have a field problem.
Feel free to contact me directly about this. I'm using a JEOL 6300F with a Vision system, and it's no problem to obtain a clean 100K digital image.
Jim Stets Air Products and Chemicals, Inc. Allentown, PA stetsjr-at-ttown.apci.com
Ye Olde Disclaimer: I speak for myself, not my employer.
I was once employed at Baylor College of Medicine in Houston, Texas in the Division of Molecular Virology. One proffessor there did many studies on Calicivirus. Her name is Mary K. Estes. I do not know her email address but it is avaiable through Baylor's Web page address'. Hope you can obtain the info you need through her.
Best of Luck, Ed Calomeni Medical College of Ohio Dept. Pathology Toledo, OH emlab-at-opus.mco.edu
Regarding the apparent drift problem in Greg Rudomen's images:
We have seen similar effects with a JEOL 6100 interfaced to a Voyager EDS/image acquisition system. The effect is also noticeable when using the image capture features of the SEM itself. Slow scans over a minute or two, whether for a Polaroid or for digitizing, do not show the problem, as the digitization occurs only once at each beam location and a small drift would be unnoticeable in terms of seeing a distorted image; however, for elemental mapping using single scans, with long dwell times resulting in acquisitions taking hours, the drift/distortion is extremely evident. Averaging, integrating or other algorithms which entail multiple scans show the diffuseness very strongly. Most frustrating is that the problem is sometimes worse during some sessions compared to others, with no obvious differences in specimens or operating conditions.
This problem was suggested by the SEM manufacturer to be one of charging or poor grounding in our specimens. Not likely, though, as we were observing Si and GaAs crystals when the problem was most noticeable in images. The problems were most notable with these because the magnifications were considerably higher than other routine work done here. Decreasing the beam energy to ca. 1-5 keV did not improve the problem, suggesting that charging plays no role.
The problem is as yet unresolved, but I wanted to add our observations. For the time being, I acquire images using slow scans only. Maybe it has something to do with imaging on every third Tuesday of months ending with the letter "y"...
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I got caught with some drift the other day on an E-SEM. The image was drifting slowly in one direction. I half suspected charging, although the drift was only in one direction and varying the pressure and cutting back the beam did not help.
The problem was eventually tracked back to the modeling clay that I had used to support the irregularly shaped sample. I was not aware that it outgassed fast enough to change dimensions in the SEM. Now I know better. ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091 E-Mail: wes-at-ameslab.gov (or: wesaia-at-iastate.edu)
coal characterization and processing electron microscopy, x-ray analysis, image analysis computer applications
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Hi all, I snipped the above lines from Ya Chen's response to Bonnie. As Nestor posted last week, there have been some difficulties with the server, and it seems that I have missed some of the posts. I hold a position at the University of Iowa's Central Microscopy Research Facility. We are a multi-user resource for microscopy related techniques. We have two SEMs that we train people to operate. ANYONE can use our facility, as long as we train them, or/and they prove themselves competent in the operation of the instrument. At that point, they are issued keys, and can reserve the instrument to suit their schedule. We have never had to ask for keys back from an individual, and any problems that have occured were corrected with additional training. We do have service contracts on our microscopes, and I can be fairly handy with a tool box. Knock on wood, we have yet to have a catastrophic event (one SEM is a field emitter). With the robust design of modern electron microscopes, I don't anticipate any, either. For more on the operating philosophy of our lab, as well as instrumentation available, I suggest you visit our website at http://www.uiowa.edu/~cemrf/.
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
Hi all, I snipped the above lines from Ya Chen's response to Bonnie. As Nestor posted last week, there have been some difficulties with the server, and it seems that I have missed some of the posts. I hold a position at the University of Iowa's Central Microscopy Research Facility. We are a multi-user resource for microscopy related techniques. We have two SEMs that we train people to operate. ANYONE can use our facility, as long as we train them, or/and they prove themselves competent in the operation of the instrument. At that point, they are issued keys, and can reserve the instrument to suit their schedule. We have never had to ask for keys back from an individual, and any problems that have occured were corrected with additional training. We do have service contracts on our microscopes, and I can be fairly handy with a tool box. Knock on wood, we have yet to have a catastrophic event (one SEM is a field emitter). With the robust design of modern electron microscopes, I don't anticipate any, either. For more on the operating philosophy of our lab, as well as instrumentation available, I suggest you visit our website at http://www.uiowa.edu/~cemrf/.
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POSTDOCTORAL POSITION (US citizens only)
The Physical Metallurgy Branch of the Naval Research Laboratory is currently looking for a postdoctoral researcher to study solid-solid phase transformations. The ideal candidate would have a strong background in both transmission electron microscopy and materials science (preferably with a concentration in physical metallurgy). Current research interests center on the study of solid-solid phase transformations in conventional and advanced metallic alloys, ranging from fundamental studies of martensite and precipitate morphologies in steels to microstructural analysis of low-carbon steel weldments, advanced shape memory alloys, new beta-titanium alloys, and metallic thin film and multilayer materials. Please contact either myself (Fonda-at-anvil.nrl.navy.mil, 202-767-2622) or Dr. George Spanos (Spanos-at-anvil.nrl.navy.mil, 202-767-5799) for further details about these and other research programs.
EQUIPMENT JEOL 200CX, Philips CM30, and Hitachi 9000 TEMs Isothermal heat treatment facility for the study of rapid phase transformations in an inert environment Quenching and deformation dilatometer Hitachi FEG-SEM Auger electron spectroscope
PROGRAM Descriptions of the postdoctoral programs at NRL may be obtained from Lesley Renfro, Program coordinator, Code 1005.7, Naval Research Laboratory, Washington, DC, 20375 (Renfro-at-utopia.nrl.navy.mil, 202-767-3865). Dr. Spanos is listed as research advisor for the NRC program descriptions.
ELIGIBILITY The postdoctoral programs at NRL are open only to US citizens.
_____________________________________________________________ Richard W. Fonda Naval Research Laboratory (202) 767-2622 Code 6324 (202) 767-2623 fax Washington DC 20375 _____________________________________________________________
Could someone explain the difference between DEPEX and DPX mounting media or are they used interchangeably? If you have a fluorescently labelled sample cleared in methyl salicylate, does anyone have a protocol for mounting in DPX? Do you have go back to xylene before mounting in DPX or can you go directly to DPX from methyl salicylate? How long does the DPX taken to harden on a 100 um sample? Thanks.
Patty Jansma Tel:602-621-6671 plj-at-manduca.neurobio.arizona.edu Arizona Research Labs Division of Neurobiology University of Arizona
The following meeting may be of interest to some readers of this listerv.
WORKSHOP
PCR-IN SITU HYBRIDIZATION: =20 What you need to know to make it work
As part of the Histochemical Society Annual Meeting,=20 August 2-3, 1996,=20 Place: Hyatt Regency Hotel, Bethesda, MD
=09Topic: The use of PCR has revolutionized study of low-abundance=20 DNA and RNA molecules and recent advances have made it possible to=20 amplify DNA and cDNA targets in fixed cells and paraffin-embedded tissues= =20 on slides. The use of these techniques is beginning to have a major=20 impact in both cell biology and diagnostic pathology. The workshop=20 will deal with: =20 =09=A5methodologies to label and detect nucleic acids using=20 non-radioactive primers=20 =09=A5advantages and disadvantages of PCR-ISH, pitfalls, controls,=20 optimal protocols, effects of fixatives, protease digestion, primers,=20 nucleotides, relevant enzymes and related issues. =20 =09=A5planned format includes lectures, question periods and=20 interactive demonstrations of in situ-PCR, PCR-in situ hybridization=20 and reverse transcriptase-in situ PCR. Thermocycling systems appropriate= =20 for processing selected tissue specimens will be utilized and will=20 generally be available during the workshop. =20 =09=A5A syllabus will be provided for workshop participants.=20
=09Organizer: G. J. Nuovo, SUNY at Stony Brook
A related symposium will be on=20
ENHANCEMENT METHODS FOR IN SITU HYBRIDIZATION=20
=09Topic: Fluorescent in situ hybridization (FISH) techniques have=20 been utilized for the past decade and offer a widely used microscopical= =20 tool providing high sensitivity and resolution, as well as the ability to= =20 detect multiple targets. To increase the sensitivity of FISH, indirect=20 amplification by immunological methods has often been the method of=20 choice. This symposium will include the presentation of several novel=20 approaches using the enzymatic deposition of fluorescent reporter=20 molecules to amplify target localization. These methods may offer a new=20 approach to increase both the detection, resolution, and sensitivity of=20 the FISH method.=20 =09Organizer: S.L. Erlandsen =09Speakers:=20 =09S. L. Erlandsen, University of Minnesota, Introduction. =09A.K. Raap, Leiden University, Sensitive and high resolution FISH=20 =09B. Schmidt, Carnegie Mellon University, Signal amplification in=20 the detection of single copy DNA + RNA by enzyme catalyzed deposition=20 (CARD) of the novel fluorescent reporter substrate, Cy 3.29-tyramide. =09V.L. Singer, Molecular Probes, Inc., The ELF alkaline=20 phosphatase substrate provides a bright, photostable, fluorescent signal=20 amplification method for FISH.=20
Another symposium will be on=20
APPLICATIONS OF MOLECULAR TECHNOLOGIES TO DIAGNOSTIC AND INVESTIGATIVE=20 PROBLEMS OF BREAST CANCER.=20
=09Topic: The symposium will include presentations on in situ=20 analysis of control of cell proliferation in breast cancer, molecular=20 markers of apoptosis and their correlation with hormone receptor=20 expression and other histologic prognostic markers and the structure of=20 breast tissue as it related to breast cancer development.=20 =09Organizer: A. M. Gown =09Speakers:=20 =09H. Wolfe, Tufts University, Molecular markers of apoptosis and=20 their correlation with hormone receptor expression and other histologic=20 prognostic markers. =09R. Jensen, Vanderbilt University, BRCA1 gene expression in breast cancer cells. =09A.M. Gown, University of Washington, In situ analysis of control=20 of cell proliferation in breast cancer. =09J.W. Gray, UC San Francisco, Applications of molecular techniques=20 to diagnostic and investigative problems of breast cancer.
POSTERS ON TOPICS RELATED TO THE WORKSHOP AND SYMPOSIA are welcome =20 (abstract deadline: March 15, 1996) and abstracts will be published as=20 Proceedings of the Histochemical Society Meeting in the Journal of=20 Histochemistry and Cytochemistry. .
CONTACT:=20 Inquiries about program matters and requests for registration materials=20 or abstract forms, and hotel accommodations should be sent to: The=20 Histochemical Society, 4 Barlows Landing Rd., Pocasset, MA 02559=20 (Telephone: 508-563-1155; FAX 508-563-1211 or e-mail: lmaser-at-mbl.edu). =20 or inquiries related to the scientific program to: Dr. W.L. Stahl=20 (e-mail: wlstahl-at-u.washington.edu).=20 =20
Dear Greg, Can you see the image drift at high mag. in TV-rate? Is this perhaps EM field ripple? or mechanical vibration? The image acquisition system can only acquire what the SEM puts out, so this drift will certainly wipe out the benefits of frame averaging. If it takes so long to acquire a frame-averaged image, why not just acquire one long frame? I have a passive system and usually take a 10 sec., 1024X768 single frame, but the image certainly shows up any drift of the SEM stage. This can always be traced to the specimen not being secured well enough. Sticky tabs are often guilty of this. If the SEM doesn't drift, the image is perfect to 50,000X.
One problem I have had in the past is bad vibration problems at 20 minutes past the hour on class days. This is when classes change and two hundred undergrad engineers tromp through the halls. People doing critical hi-res, low kV work know to do it after hours, when the building is quieter and the elevators aren't running. Good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
I am looking for a mounting medium which can be used for coverslipping Giemsa stained cells containing latex microspheres. Xylene containing mounting media cannot been used because of the solubility of the latex microspheres. On the other hand, water soluble mounting media, like gelatin/glycerin, destain the Giemsa. The staining is important to me to clearly locate (for counting) the microspheres within the cell borders. Counting without coverslip can be done but I prefere using a 40x/1.0 objective (oil).
I'm looking for other microscope pools, where teaching microscopes are shared to maximize usage. I've worked for 20 years at Microscope Services at the University of California, Davis. We have about 2100 optical microscopes of high quality, and move 600-700 around every quarter from classrooms where they aren't needed to those where they are. We also provide preventive maintenance and repairs, short courses microscopes for research, recharging for our services. This arrangement works well at UC Davis, so it must be duplicated at other schools. I just don't know where. If any of you know of schools with similar arrangements, would you please post a contact for me? I would very much like to hear from someone with similar tasks and problems. Richard L. Markgraf Microscope Services University of California, Davis Ph. (916)752-3477 Fax (916)752-6363 rlmarkgraf-at-ucdavis.edu
Joint Annual Meeting of the Microscopy Society of America, Microbeam Analysis Society and the Microscopical Society of Canada/Societe de Microscopie du Canada, August 11-15, 1996, Minneapolis, Minnesota.
Deadline for receipt of Extended Abstracts: March 15, 1996.
Approximately 7,000 Registration Bulletin / Call for Papers packages have been mailed, most on February 2, to: the memberships of the three Sponsoring Societies; the memberships of four Local Affiliates of the Microscopy Society of America (Northwestern Ohio Microscopy Society, Minnesota Microscopy Society, Southern California Society for Electron Microscopy - Technologists, and the Iowa Microscopy Society); individuals who attended Microscopy & Microanalysis '95, Kansas City; and other individuals who have requested one.
Please contact our office if you should have received the Registration Bulletin / Call for Papers but haven't, or if you would like to. We'll respond to your request immediately. Please note that the deadline for receipt of Extended Abstracts is March 15, 1996.
Hope to see you in Minneapolis,
Larry Maser
Microscopy & Microanalysis '96 4 Barlows Landing Rd., Suite 8 Pocasset MA 02559
Message-Id: {199602161628.KAA13875-at-mailhub.iastate.edu} X-Sender: wes-at-pop.ameslab.gov X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
From another university lab:
We too allow users on the scope after training as many in academic settings appear to do. I routinely query the potential users about the realistic duration of their work as we have trained some at their insistence only to find out they have no specific project in mind, or that their actual SEM usage was not nearly what they first imagined. Of course, since they are charged for operator assistance, I guess we will continue to train them if they care to pay for it.
Now a question, We have adopted the practice of leaving the filament current control on our JEOL 840A at the proper saturation point (for our normal 15 kv operating point) and just turning off the high voltage control for exchanging samples. We have thus avoided problems from inexperienced operators over-saturating. It appears to be working as our filament lifetimes have increased to the values for when only experienced users operated the scope. My question to the list is whether there might be any adverse consequences from this practice as there do not appear to be any.
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Hello All, A graduate student here has been given the task of looking at the surface of a highly polished stainless steel ball which is used in hip replacements. The ball is about 2.5-3 cm in diameter and as you might imagine, is extremely reflective. Since all I have ever examined in an SEM are biological samples, I haven't been able to help her very much. This morning we got what appeared to be a grid image on the surface and I am wondering if this might not be a reflection of the interior surface of the specimen chamber. I suggested sputter coating the surface to cut down on reflection and mounting it on some double stick tape to keep it stable; but she doesn't think the person who assigned her this task would want that. Without regard to the wants of the person assigning the task, do any of you in the materials science have any suggestions which might be helpful to this young lady. She and I would both be very grateful. TIA, Sandra Zane Sandra F. Zane, EM Tech. sfzane-at-unccvm.uncc.edu Dept of Biol., UNC Charlotte (704) 547-4051 9201 University City Blvd. Fax (704) 547-3128 Charlotte, NC 28223-0001
} Hi Everyone, } I have a JEOL 5300 SEM to which a beam controlled image } acquisition system has been added. Images are acquired at } 640x512 by frame averaging. It takes 1min 20sec. to } collect five frames. This is about the same time it takes } to do a Polaroid. } Here is the problem. At magnifications above 5000x the } images are fuzzy due to some kind of drift. At lower mags } the drift is not noticeable. Polaroids do not show the } drift. The drift rate is 100-500u per five minutes. I } have the SEM manufarturer working on this too. So far I } have not gotten acceptable results } Now for the questions. Has anyone had this problem on } other image acquisition systems? Am I trying to do } something that can't be done due to drift that all SEMs } have to some degree? } Thanks for your answers in advance. } } Gregory Rudomen } Greg-at-umic.umic.sunysb.edu } University Microscopy Imaging Center } S.U.N.Y. at Stony Brook }
Gregory, There are 3 kinds of averaging techniques used in SEM digitization: frame averaging, line averaging and pixel averaging. For most of "built-in" acquisition system, the frame capture is used, so only support frame averaging. The drawback for using frame averaging is to produce a fuzzy image as long as drafting exist. Pixel averaging is mostly suitable to overcome this situation.
Ya Chen
Ya Chen Cryo/SEM project Coordinator TEL : 608-263-8481 Integrated Microscopy Resource (IMR) FAX : 608-265-4076 University of Wisconsin-Madison Email:YChen-at-macc.wisc.edu IMR WWW Home Page: http://www.bocklabs.wisc.edu/imr/imr.html IMR Symposium on integrated microscopy '96: Sept. 20-22, 1996
Dear Sandra, It is very difficult to look at the surface of any highly-polished metal, since there is nothing to focus on. However, you cannot get a reflection image in an SEM except by charge buildup, so if you did get a grid image from your secondary detector, it implies the ball is not well grounded. If you stick it down, use conductive sticky-tabs or make a connection useing silver or carbon dag. One help to focusing is to draw on the top surface with a felt-tip pen, so you can find the initial focus. My experience though, is that a good reflextive surface gives you very little to see, even at high mag. Good luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
} Now a question, } We have adopted the practice of leaving the filament current control on our } JEOL 840A at the proper saturation point (for our normal 15 kv operating } point) and just turning off the high voltage control for exchanging samples. } We have thus avoided problems from inexperienced operators over-saturating. } It appears to be working as our filament lifetimes have increased to the } values for when only experienced users operated the scope. My question to } the list is whether there might be any adverse consequences from this } practice as there do not appear to be any. Dear Warren, We always leave the filament current up to the saturation point (or just under it for maximum filament life). In fact, to stop little fingers, I removed the filament current knob from my console years ago. I can change it with a screwdriver when I need to. I always turn the acc. voltage on and off and I get at least a month out iof a filament at 20kv. I don't know if I would do it on my 200kv TEM, but SEM is fine. Best of luck, Mary
Mary Mager Electron Microscopist Metals and Materials Eng, UBC 309 - 6350 Stores Rd. Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648, fax: 604-822-3619 email: mager-at-unixg.ubc.ca
} The maximum energy available to impart to an atom in the bulk is given by the } expression, } } E = E0 * 4* (Mm)/(M+m)^2, } } where M is the bulk atom mass, m is the ion mass and E0 is the ion energy. } } It seems to me that if you have sufficient energy for removal of ions, that you } will always exceed the minimum threshold energy for displacent of atoms.
......... Not true!
The sputtering rate/cross-section at the SURFACE of a material is not the same as the displacement rate/cross-section in the bulk. The simple way to think about this is that a surface you only have ~ 1/2 the number of bonds that you have in the interior of a bulk. Thus the energy needed to remove a surface atom is LESS than that needed to remove one from the bulk. There is extensive literature on this unfortunatley, I don't have the references readily at hand here at home.
Charlie Bradley and I did work on this at ANL (for case of electron sputtering) back in the late 80's some of which was published in the (E)MSA proceedings. There is also an ANL Technical Report which tabulates both sputtering and displacement cross-sections for the entire periodic table. The computer programs which do the calculations for this are in the Software Library which is accessible by anonymous ftp at
ftp.msa.microscopy.com
they are in the path /pub/3-MMSLib/HVEM
look for the files on DMottCS, TotCS, Recoil. Like I said, these programs were written for the case of electron sputtering, however, they could be modified for the ion-beam case, if one were sufficiently interested. At any rate, you will not displace the atoms in the bulk, but only the surface and near suface atoms.
There is another program called TRIM which is used by the Ion-Beam Analysis community to calculate bulk displacements in solids by Ions. I'll dig up the references and post them next week when I get back into the lab.
-- [ From: Garber, Charles A. * EMC.Ver #2.10P ] --
In response to:
} I'd like some input, opinions etc. on the pitfalls of having multiple } operators on an SEM.
During the years 1963-1967, at what was then Case Institute (now CWRU), during the conduct of my MS and Ph. D. thesis work, I did with my own two hands, all of my own TEM work which at times involved in excess of thirty hours instrument time per week over extended periods of time. As a lowly (yes, we were definitely made to feel lowly) graduate student, to this day, I remember how I had to tip-toe around the manager of the EM facility. All of the graduate students had their own keys both to the EM lab and also the darkroom, but one snap of the manager's fingers could have keys snatched away in an instant. And for the entire four years of my career as a graduate student, I did have my own keys and for the most part, that part of my thesis work requiring EM was done between the hours of 6:00 pm to 12:00 midnight (often times later), and without supervision. Indeed virtually any graduate student who had need for EM work was issued keys and for the most part, their work was done during these "off hours" as well.
Indeed, to have the experience of actually being responsible for the health and well-being of the instrument, and to be held accountable for it, I always considered an important part of my own graduate experience and education.
And sitting there, night after night, peering at the green image on the screen, and experiencing that electrifying feeling that comes only when seeing something for the first time, something perhaps no one else has previously seen, was a part of my own life I will never forget. The challenge of proving first to myself and then to others that what I thought I had seen was in fact "real" and was not an "artifact" has had a lasting positive impression on my own life and my own career for that matter. After all, how else can one learn to be their own worst (best?) "devil's advocate" if they have not done the work themselves?
How else can one "sell others" on the idea that what they have indeed seen is real and not some preparation artifact? How many times have I listed to presentations by one of those arm chair EM people and when asked a question, could not even tell whether a sample was gold coated or not! Or with what a TEM sample might have been stained. Not too long ago I witnessed a speaker not being able to answer whether a micrograph was by SEM or TEM. Such responses hardly evoke high levels of confidence in someone else's conclusions.
Yet I can also remember some students who did not have research projects depending that much on EM, perhaps their use of EM being confined to closing a few lose ends, and where the inclusion of some EM work was just a small part of their overall project. Indeed, either because of less interest in doing it with their own two hands, or perhaps other reasons, these students had that laboratory manager, the one who made us jump through hoops, do the work, during daylight hours with the student peering over his shoulders. On the other hand, there were those who did have minimal need for EM work, but underwent the extensive training voluntarily and with enthusiasm as part of their overall graduate training experience.
I appreciated that lack of bureaucracy that existed in those days, and the climate was such that the highest levels of curiosity were encouraged. There is no question that if I was not able to have done my own work, in the way that I did it, yes, perhaps even late at night when there were fewer building vibrations and toilets flushing, I would be a much different person. After all, at 1:00 am, when something does go wrong, to deal with it, on your own, responsibily, to make judgemental decisions, with no one else to turn to, is in itself an important part of the educational process.
And what about down time? Of course there was the normal down time and probably there was more down time than might have other wise been the case because there were multiple users. But it was never an excessive amount of downtime. And these were during the times when instruments were not engineered like they are today, and if anything, relative to modern instruments, the amount of downtime experienced should have inherently been significantly more than would be the case today. But excessive downtime never happened. Indeed seeing the way some of these judgemental decisions were handled in those days has been valuable in the management of my own analytical laboratory services business today.
I have followed the discussion on this listserver. I am really quite surprised with the bureaucracies that seem to have evolved to somehow "protect" the equipment from mere ordinary "mortals". What happened to the ability to use the same kind of good judgment that abounded in the EM laboratories of thirty years ago? Of course it is always easier to go by way of a script and never deviate. Go by the book. After all, no decision making has to be done at all. But something very very valuable and fundamental is lost when those with the interest are not able to sit down and take their own data, at their own speed, and on their own terms.
I can appreciate the concern about minimizing downtime more than most people, in my case, such costs come right out of my own pocket. It is a very real concern. However, be it in industry or academia, give a lot of thought before you shut out that inquisitive researcher who just might make that big discovery that moves us all forward another notch. A well managed industrial laboratory should have made provision for emergency scope access anyhow so even if there was a bit of additional downtime, surely that can not be thought of as throwing a monkey wrench into that company's ability to have an instrument available to work on solving emergency plant problems. If you have an instrument that is that sensitive to multiple users, perhaps the time has come to look at insturments made by some other manufacturer.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
At South Bay Technology we manufacture a Single Vertical Jet Electropolisher which can be used for either electrolytic or chemical polishing. It is possible to use it with Br-Methanol solutions and it does offer automatic termination as you requested. It is also offers an automatic rinse option and several other features. If you would like information on this product or on any of our other sample preparation products, please feel free to contact me off-line. If you have an interest, I can also put you in touch with our distributor in Spain.
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David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com sbt-at-msa.microscopy.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
Message-ID: {199602171916.OAA06508-at-IndyNet.indy.net} To: "Sandra F. Zane" {sfzane-at-unccvm.uncc.edu} , Microscopy list {microscopy-at-Sparc5.Microscopy.Com}
-- [ From: Paul E. Fischione * EMC.Ver #2.3 ] --
To expand on the ion energy vs. damage discussion, we have found that by reducing the ion energy, a decrease in ion damage is experienced at a given angle. During our research effort in developing the Hollow Anode Discharge (HAD) ion source, we concentrated on achieving a broad range of ion energy operation. Currently, the HAD source will operate with a minimum voltage (accelerating) of between 500 and 800 volts and produce ion energies on the order of a few microamps/square cm. At the maximum energy (6 kV), ion current is in excess of 400 microamps/sq.cm resulting in very high milling rates which are preferable when milling at low angles.
As with any preparation procedure, the key is defining the proper parameters. Having a versatile ion mill that can effectively mill at low/high angles, low/high voltages, and with liquid nitrogen cooling affords the required flexibility.
There are some good references in the MRS Specimen Preparation Proceedings.
Regards,
Paul Fischione E.A. Fischione Instruments, Inc. 9003 Corporate Circle Export, PA 15632 Phone (412)325-5444 FAX (412)325-5443 e-mail paul.fischione-at-internetmci.com
I would like to express very strong agreement with the sentiments of Chuck Gabor; certainly within the academic setting protecting microscopes from students is counterproductive. One thing I will argue is a credit to our current system here is that we allow undergraduates to have hands on experience using SEM's and TEM's. While it is true that there are dangers of breakages, so what! To my knowledge almost all the "big names" in electron microscopy made stupid mistakes at one time or another - one example that I know of involved switching of the vacuum and water lines on a TEM.
Message-ID: {MAPI.Id.0016.00683536202020203843394130303030-at-MAPI.to.RFC822} Read-Receipt-To: Jean-Louis Beek {ah56-at-solo.pipex.co.za} Priority: Normal To: microscopy-at-Sparc5.Microscopy.Com MIME-Version: 1.0
} There is a virus being sent about through the email system, it is } usually sent with the subject (good news....). I received the below mail } from my brother, who works with ICL. ICL sent the below message to all } of it's employees. } } Please see the attached and take appropriate action. } } } Issued by IT Services } } } COMPUTER VIRUS } } There is a computer virus that is being sent across the Internet. If } you receive an e-mail message with the subject line "Good Times," DO } NOT read the message, DELETE it immediately. Please read the messages } below. } } Some miscreant is sending e-mail under the title "Good Times" } nationwide. If you get anything like this, DON'T DOWNLOAD THE FILE! } It has a virus that rewrites your hard drive, obliterating anything } on it. } } Please be careful and forward this mail to anyone you care about. } } ************************************************************* } } WARNING!!!!!!! INTERNET VIRUS } } The FCC released a warning last Wednesday concerning a matter of major } importance to any regular user of the Internet. Apparently a new } computer virus has been engineered by a user of AMERICA ON LINE that } is unparalleled in its destructive capability. Other more well-known } viruses such as "Stoned," "Airwolf" and "Michaelangelo" pale in } comparison to the prospects of this newest creation by a warped } mentality. What makes this virus so terrifying, said the FCC, is the } fact that no program needs to be exchanged for a new computer to be } infected. It can be spread through the existing e-mail systems of the } Internet. Once a computer is infected, one of several things can } happen. If the computer contains a hard drive, that will most likely } be destroyed. If the program is not stopped, the computer's processor } will be placed in an nth-complexity infinite binary loop - which can } severely damage the processor if left running that way too long. } Unfortunately, most novice computer users will not realize what is } happening until it is far too late. Luckily, there is one sure means } of detecting what is now known as the "Good Times" virus. It always } travels to new computers the same way in a text e-mail message with } the subject line reading "Good Times." Avoiding infection is easy } once the file has been received - do not read it! The act of loading } the file into the mail server's ASCII buffer causes the "Good Times" } mainline program to initialize and execute. } } The program is highly intelligent - it will send copies of itself to } everyone whose e-mail address is contained in a receive-mail file or a } sent-mail file, if it can find one. It will then proceed to trash the } computer it is running on. The bottom line here is - if you receive a } file with the subject line "Good Times," delete it immediately! Do } not read it! Rest assured that the name was on the "From" line was } surely struck by the virus. Warn your friends and local system users } of this newest threat to the Internet! It could save them a lot of } time and money. } } E N D O F N O T E } } -- } Tomaz Pereira Garcez (u01tp-at-abdn.ac.uk) +44 1224 632549
Diana van Driel Dept Ophthalmology C09 Sydney University 2006 NSW, AUSTRALIA
From the various symptoms that have been described I think that Rick Mott's last suggested source of this problem is most likely to be correct. In conventional scanning the scan is usually synchronized to the line frequency to minimize distortions resulting from stray fields. When using an external scan generator for acquiring digitized images some of the scan speeds will be asynchronous with the line frequency. If there are stray fields in the vicinity of the SEM these will then cause some sort of image distortion on the screen, often showing as a uniformly jagged appearance of what should be straight edges. If one then averages a number of such frames it could lead to what has been described as a fuzzy image.
In order to establish if this is the problem it will be necessary to test for stray A/C fields around the SEM. These usually arise from sources such as large power supply cables running under the floor or in a nearby wall, but as Rick Mott has suggested, could simply be a problem of bad or wrongly routed earthing. If one doesn't have access to a proper magnetic field testing coil an old EM lens coil connected to an oscilloscope, although not calibrated, nevertheless does very well for comparing one area with another for frequency and magnitude of fields.
Good luck!
Robin Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za (tel: +27 461 318168 - fax: +27 461 24377)
I have run an EM service unit for over 25 years during time which the question of hands-on usage of the instruments has often been discussed. This unit's size (4 EM's and a staff of four) is probably common to many throughout the world and from what I have seen of the present discussion on this issue our problems/experiences/solutions are similarly shared by many people elsewhere.
While there is no doubt that instruments will be better off if only operated by trained full time EM Unit staff this policy is seldom practical. Policies in force 25 years ago, when it usually required the skills of regular operators to get the best out of electron microscopes, no longer need to be so rigidly applied. Today, not only is it easier for practically anyone to get good results out of modern EM's with a minimum of training but also it is much less likely that an inexperienced user will cause any serious damage (with instruments such as our JEOL 1210, filament saturation is pre-programmed and even if finger-happy users create havoc with the beam the correct alignment can be fully restored in one simple operation). We have found that to work effectively the policy needs to be flexible and dependent largely on two important factors:
- the nature and extent of the work - the aptitude of the user for using this instrumentation.
In very general terms the policy successfully applied here is that if a user's project is of the "once-off" variety (likely to involve only one or two sessions) we will carry out or closely supervise almost everything involving our equipment, but at the same time encourage the user to share in the viewing, interpretation and photography of images, etc. If the project will require repeated use of the EM (more than, say, 3 or 4 sessions) then we find it worth our while to train that user to operate the instrument more-or-less by themselves. The extent to which this training extends depends on the anticipated amount of usage and the perceived ability of that user to handle these instruments. What generally happens is that the longer that person uses the instrument the more procedures he/she will be taught to perform. For a while this operation is confined to just viewing and micrography with assistance being given in other necessary operations such as specimen and film exchange. Seldom, however, does user training extend to anything beyond actual operation procedures (specimen exchange, viewing and micrography). Although specimen preparation by properly trained people is routinely allowed after hours in the EM Unit, use of the EM's is very seldom permitted when no members of the EM Unit staff are in the vicinity, mainly because of the validity of the university's insurance cover (both for the user and the instruments) in the event of an emergency under such circumstances. Maintenance operations such as start- up, alignment, filament exchange, etc, are not carried out by users.
Although it is sometimes seen to be discriminatory to have one policy for one group of users and another for others, as soon as the rationale behind the policy has been made clear we have seldom had any further problems. As a result this compromise policy works well and as long as our circumstances remain as they are I see no reason to make any significant changes.
Robin Cross Director : EM Unit, Rhodes University, Grahamstown, South Africa eurc-at-giraffe.ru.ac.za (tel: +27 461 318168 - fax: +27 461 24377)
} I have recently purchased an old Carl Zeiss Jena compound microscope , =
While on the subject, we have a Jenavert microscope that we inherited that is in need of some parts to maintain tension in the stage adjustment. Our local service provider has been waiting 6 months for these parts. Anyone have any news about when we can expect to see our parts? How about any news about Carl Zeiss Jena itself?
Tom Ostertag ostertag_tom-at-mn15-gw.mavd.honeywell.com Tom.Ostertag-at-mavdmh.honeywell.com tostertag-at-tcm.mn.org Minneapolis, MN
Mr-Received: by mta PETVAX.MUAS; Relayed; Mon, 19 Feb 1996 08:33:01 -0400 Mr-Received: by mta PETVAX; Relayed; Mon, 19 Feb 1996 08:33:03 -0400 Mr-Received: by mta SRVR01; Relayed; Mon, 19 Feb 1996 08:40:44 -0400 Disclose-Recipients: prohibited MSA Microscopy Mailing List {MICROSCOPY-at-Sparc5.Microscopy.Com} Message-Id: {8601330819021996/A02465/PETVAX/11A29A210000*-at-MHS} X-Envelope-To: dianavd-at-eye.usyd.edu.au, MICROSCOPY-at-MSA.Microscopy.com Autoforwarded: false Mime-Version: 1.0 Content-Type: TEXT/PLAIN; CHARSET=US-ASCII Content-Transfer-Encoding: 7BIT Importance: normal Priority: normal Sensitivity: Company-Confidential Ua-Content-Id: 11A29A210000 X400-Mts-Identifier: [;8601330819021996/A02465/PETVAX] Hop-Count: 2
The Good Times Virus threat is a complete fabrication that was started by some members of America Online. Members should completely disregard posted messages that perpetuate this myth that has assumed a life of it's own.
Optical reflectivness make no sense for SEM. Metallic samples are almost ideal for SEM observation. I suppose, the only problem you have is focusing on specimen which is about couple cm above stage surface. Try using full focus range at minimum magnification.
Vladimir Dusevich dusevich-at-astro.ocis.temple.edu
On Fri, 16 Feb 1996, Sandra F. Zane wrote:
} Hello All, } A graduate student here has been given the task of looking at the } surface of a highly polished stainless steel ball which is used in hip } replacements. The ball is about 2.5-3 cm in diameter and as you might } imagine, is extremely reflective. Since all I have ever examined in an SEM } are biological samples, I haven't been able to help her very much. This } morning we got what appeared to be a grid image on the surface and I am } wondering if this might not be a reflection of the interior surface of the } specimen chamber. I suggested sputter coating the surface to cut down on } reflection and mounting it on some double stick tape to keep it stable; but } she doesn't think the person who assigned her this task would want that. } Without regard to the wants of the person assigning the task, do any of you } in the materials science have any suggestions which might be helpful to this } young lady. She and I would both be very grateful. } TIA, Sandra Zane } Sandra F. Zane, EM Tech. sfzane-at-unccvm.uncc.edu } Dept of Biol., UNC Charlotte (704) 547-4051 } 9201 University City Blvd. Fax (704) 547-3128 } Charlotte, NC 28223-0001 } } } }
A question for those doing thin section TEM for diagnostic pathology:
Is there a standard protocol for rapid processing of tissues where overnight ( or sooner) results are required? I am talking straight morpholgy. -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at- Greg Erdos Phone: 352-392-1295 Scientific Director, ICBR Electron Microscopy Core Lab 218 Carr Hall Fax: 352-846-0251 University of Florida E-mail: gwe-at-biotech.ufl.edu Gainesville, FL 32611 -at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at--at-
As most of you know, when Germany was reunited so was Zeiss aus Jena and Zeiss aus Oberkochen. They are now one company. They are gradually moving to East Germany. Nina Allen
On 19 Feb 1996, Ostertag Tom wrote:
} Jean-Louis Beek wrote: } } } I have recently purchased an old Carl Zeiss Jena compound microscope , = } } While on the subject, we have a Jenavert microscope that we inherited that is } in need of some parts to maintain tension in the stage adjustment. Our local } service provider has been waiting 6 months for these parts. Anyone have any } news about when we can expect to see our parts? How about any news about Carl } Zeiss Jena itself? } } Tom Ostertag } ostertag_tom-at-mn15-gw.mavd.honeywell.com } Tom.Ostertag-at-mavdmh.honeywell.com } tostertag-at-tcm.mn.org } Minneapolis, MN } } } } } } }
Message-Id: {199602191518.JAA09603-at-mailhub.iastate.edu} X-Sender: wes-at-pop.ameslab.gov X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Personally, my taste in humor runs in a different direction.
The "Good times virus" is a hoax. It has been around for several years now. About once a month I hear about it again. And as often, those who know better point out that it is a hoax. The most damage that comes from it is the cluttering of the mail system with its false alarm and the clarifications that it is a fake.
Viruses have to be executed, not just downloaded, to infect a machine. Therefore, it is a good idea not to automatically open, execute, or run attachments. Our "convenient" mail programs with their auto-this and that features could do us a disservice.
The Macro viruses ARE making the rounds. These viruses hide out in autoexecute macros in Word and Excel documents. Lest you think Microsoft alone has a problem, any software that has such a macro capability is potentially at risk. Still the document has to be opened for the macros to execute.
I continue to see warnings about AOL-Gold and PKZip 3.0 viruses. I have not seen them personally, but have been informed by reliable sources (CICA, et al) that they are legitimate. PKZip's current version is 2.04; the 3.0 designation is bogus and carries the virus. AOL-Gold has a virus hidden in the alleged install procedure. I thought AOL had pretty well stomped it out and warned their subscribers. ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091 E-Mail: wes-at-ameslab.gov (or: wesaia-at-iastate.edu)
coal characterization and processing electron microscopy, x-ray analysis, image analysis computer applications
I would ignore this it is a resurfacing of a hoax from last year.
At 11:07 AM 2/19/96, Diana van Driel wrote: } } There is a virus being sent about through the email system, it is } } usually sent with the subject (good news....). I received the below mail } } from my brother, who works with ICL. ICL sent the below message to all } } of it's employees. } } } } Please see the attached and take appropriate action. } } } } } } Issued by IT Services } } } } } } COMPUTER VIRUS } } } } There is a computer virus that is being sent across the Internet. If } } you receive an e-mail message with the subject line "Good Times," DO } } NOT read the message, DELETE it immediately. Please read the messages } } below. } } } } Some miscreant is sending e-mail under the title "Good Times" } } nationwide. If you get anything like this, DON'T DOWNLOAD THE FILE! } } It has a virus that rewrites your hard drive, obliterating anything } } on it. } } } } Please be careful and forward this mail to anyone you care about. } } } } ************************************************************* } } } } WARNING!!!!!!! INTERNET VIRUS } } } } The FCC released a warning last Wednesday concerning a matter of major } } importance to any regular user of the Internet. Apparently a new } } computer virus has been engineered by a user of AMERICA ON LINE that } } is unparalleled in its destructive capability. Other more well-known } } viruses such as "Stoned," "Airwolf" and "Michaelangelo" pale in } } comparison to the prospects of this newest creation by a warped } } mentality. What makes this virus so terrifying, said the FCC, is the } } fact that no program needs to be exchanged for a new computer to be } } infected. It can be spread through the existing e-mail systems of the } } Internet. Once a computer is infected, one of several things can } } happen. If the computer contains a hard drive, that will most likely } } be destroyed. If the program is not stopped, the computer's processor } } will be placed in an nth-complexity infinite binary loop - which can } } severely damage the processor if left running that way too long. } } Unfortunately, most novice computer users will not realize what is } } happening until it is far too late. Luckily, there is one sure means } } of detecting what is now known as the "Good Times" virus. It always } } travels to new computers the same way in a text e-mail message with } } the subject line reading "Good Times." Avoiding infection is easy } } once the file has been received - do not read it! The act of loading } } the file into the mail server's ASCII buffer causes the "Good Times" } } mainline program to initialize and execute. } } } } The program is highly intelligent - it will send copies of itself to } } everyone whose e-mail address is contained in a receive-mail file or a } } sent-mail file, if it can find one. It will then proceed to trash the } } computer it is running on. The bottom line here is - if you receive a } } file with the subject line "Good Times," delete it immediately! Do } } not read it! Rest assured that the name was on the "From" line was } } surely struck by the virus. Warn your friends and local system users } } of this newest threat to the Internet! It could save them a lot of } } time and money. } } } } E N D O F N O T E } } } } -- } } Tomaz Pereira Garcez (u01tp-at-abdn.ac.uk) +44 1224 632549 } } Diana van Driel } Dept Ophthalmology C09 } Sydney University 2006 } NSW, AUSTRALIA
John Mansfield North Campus Electron Microbeam Analysis Laboratory 413 SRB, University of Michigan 2455 Hayward, Ann Arbor MI 48109-2143 Phone: (313)936-3352 FAX (313)936-3352 jfmjfm-at-engin.umich.edu, John.F.Mansfield-at-umich.edu or jfmjfm-at-umich.edu they all reach me! URL: http://emalwww.engin.umich.edu/people/jfmjfm/jfmjfm.html
Can anyone suggest beam-stable specimens consisting of randomly-oriented 2D (i.e. molecule-thick) crystals for HREM (and other kinds of high resolution) imaging? Simulations we've done (cf. Jun'95 Image of the Month at {http://newton.umsl.edu/stei_lab/} ) suggest that they will give some interesting contrast edge-on, even without sub-2A point resolution! I am particularly interested in getting a TEM specimen of some large polycyclic aromatic hydrocarbons (PAHS), like dicoronene, for comparison with images of "2D graphite" in the cores of presolar interstellar graphite onions (cf. Jan'96 Image of the Month, ibid.).
If anyone has any ideas how one might affect an abrupt transition from 2D to 3D graphitic growth in the environs just outside the surface of a carbon-rich red giant, I would love to hear them as well.
Cheers. /philf :)
//\/\/\/\---} // Phil Fraundorf Physics&Astronomy/CME 314-5165044 philf-at-newton.umsl.edu \\ B503 U.Missouri-SL St.Louis MO 63121 USA http://newton.umsl.edu/~philf \\/\/\/\/\/\/\/---}
We were given a Sorval MT-2 ultramicrotome which we have repaired and are making ready for use. We need to locate a specimen chuck. Can anyone help? Many thanks.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Looking for TEM related courses, seminars to attend for beginner to advanced other than MAS and Scanning Inc. Use and applications to Biological TEM Biological Applications, specimen prep etc as related to TEM Immunocytochemistry, autoradiography, etc. as related to TEM
Any and all leads would be appreciated.
Thanks Greg2NJ-at-AOL.com Gregory Argentieri Sandoz Pharmaceuticals Corp Electron Microscopy Lab East Hanover, NJ 07936 201-503-8617
This thing is the vampire of the net, it keeps showing up, sucking up bandwidth and frightening people. I'd like to drive a virtual stake through the heart of the individual or group that began this. The attachment to this letter, I don't know if attachments show up on the server, has some good information sources for virus information on this hoax and in general.
Where ever I go, there I am. Bob Lawrence
I know most of you know this, but you might like the phone #s
{ISWORLD-at-IRLEARN.BITNET}
Here is another view on the supposed e-mail virus circulating the internet. Read through the forwarding notes to the CIAC newsletter.
--Jim
P.S. At this time of the year many of us accept student projects via diskette or FTP. This latest virus scare provides us with a good reminder that we should ALWAYS virus check anything we get off the net or from our students.
---------- Forwarded message ---------- } Reply-To: karyn-at-cheetah.llnl.gov } Originator: ciac-notes-at-cheetah.llnl.gov } Sender: ciac-notes-at-cheetah.llnl.gov } Precedence: bulk } From: Karyn Pichnarczyk {karyn-at-cheetah.llnl.gov} } To: ciac-at-ncar.ucar.edu } Subject: CIAC Notes 94-04 } X-Listprocessor-Version: 6.0b -- ListProcessor by Anastasios Kotsikonas } } } U.S. DOE's Computer Incident Advisory Capability } ___ __ __ _ ___ __ __ __ __ __ } / | /_\ / |\ | / \ | |_ /_ } \___ __|__ / \ \___ | \| \__/ | |__ __/ } } Number 94-04 December 6, 1994 } } ------------------- A - T - T - E - N - T - I - O - N ------------------- } | CIAC is available 24-hours a day via its two skypage numbers. To use | } | this service, dial 1-800-759-7243. The PIN numbers are: 8550070 (for | } | the CIAC duty person) and 8550074 (for the CIAC manager). Please keep | } | these numbers handy. | } ------------------------------------------------------------------------- } } Welcome to the fourth issue of CIAC Notes! This is a special edition to } clear up recent reports of a "good times" virus-hoax. Let us know if you } have topics you would like addressed or have feedback on what is useful and } what is not. Please contact the editor, Allan L. Van Lehn, CIAC, } 510-422-8193 or send E-mail to ciac-at-llnl.gov. } } $-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$ } $ Reference to any specific commercial product does not necessarily $ } $ constitute or imply its endorsement, recommendation or favoring by $ } $ CIAC, the University of California, or the United States Government.$ } $-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$-$ } } THE "Good Times" VIRUS IS AN URBAN LEGEND } } In the early part of December, CIAC started to receive information requests } about a supposed "virus" which could be contracted via America OnLine, simply } by reading a message. The following is the message that CIAC received: } } --------------------------------------------------------------------------- } | Here is some important information. Beware of a file called Goodtimes. | } | | } | Happy Chanukah everyone, and be careful out there. There is a virus on | } | America Online being sent by E-Mail. If you get anything called "Good | } | Times", DON'T read it or download it. It is a virus that will erase your | } | hard drive. Forward this to all your friends. It may help them a lot. | } --------------------------------------------------------------------------- } } THIS IS A HOAX. Upon investigation, CIAC has determined that this message } originated from both a user of America Online and a student at a university } at approximately the same time, and it was meant to be a hoax. } } CIAC has also seen other variations of this hoax, the main one is that any } electronic mail message with the subject line of "xxx-1" will infect your } computer. } } This rumor has been spreading very widely. This spread is due mainly to the } fact that many people have seen a message with "Good Times" in the header. } They delete the message without reading it, thus believing that they have } saved themselves from being attacked. These first-hand reports give a false } sense of credibility to the alert message. } } There has been one confirmation of a person who received a message with } "xxx-1" in the header, but an empty message body. Then, (in a panic, because } he had heard the alert), he checked his PC for viruses (the first time he } checked his machine in months) and found a pre-existing virus on his machine. } He incorrectly came to the conclusion that the E-mail message gave him the } virus (this particular virus could NOT POSSIBLY have spread via an E-mail } message). This person then spread his alert. } } As of this date, there are no known viruses which can infect merely through } reading a mail message. For a virus to spread some program must be executed. } Reading a mail message does not execute the mail message. Yes, Trojans have } been found as executable attachments to mail messages, the most notorious } being the IBM VM Christmas Card Trojan of 1987, also the TERM MODULE Worm } (reference CIAC Bulletin B-7) and the GAME2 MODULE Worm (CIAC Bulletin B-12). } But this is not the case for this particular "virus" alert. } } If you encounter this message being distributed on any mailing lists, simply } ignore it or send a follow-up message stating that this is a false rumor. } } Karyn Pichnarczyk } CIAC Team } ciac-at-llnl.gov } } } ------------------------------ } Contacting CIAC } } If you require additional assistance or wish to report a vulnerability, call } CIAC at 510-422-8193, fax messages to 510-423-8002 or send E-mail to } ciac-at-llnl.gov. For emergencies and off-hour assistance, call 1-800-SKY-PAGE } (759-7243) and enter PIN number 8550070 (primary) or 8550074 (secondary). } The CIAC Duty Officer, a rotating responsibility, carries the primary } skypager. The Project Leader carries the secondary skypager. If you are } unable to contact CIAC via phone, please use the skypage system. } } ------------------------------ } This document was prepared as an account of work sponsored by an agency of } the United States Government. Neither the United States Government nor the } University of California nor any of their employees, makes any warranty, } express or implied, or assumes any legal liability or responsibility for the } accuracy, completeness, or usefulness of any information, apparatus, product, } or process disclosed, or represents that its use would not infringe privately } owned rights. Reference herein to any specific commercial products, process, } or service by trade name, trademark, manufacturer, or otherwise, does not } necessarily constitute or imply its endorsement, recommendation or favoring } by the United States Government or the University of California. The views } and opinions of authors expressed herein do not necessarily state or reflect } those of the United States Government or the University of California, and } shall not be used for advertising or product endorsement purposes. } } ------------------------------ } End of CIAC Notes Number 94-04 94_12_06 } **************************************** } } } } } ------ Forwarded by FPSJQ-at-CUVMC.
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For years I have been using a protocol that allows me to section materials the next day. It is modified from a booklet by Millonig:
Fix in buffered GA (3-4 hrs), rinse (45 min), osmicate (2 hrs), rinse and dehydrate in acetone (3 hrs), infiltrate and embed in Spurr's (about 1 hr). Instead of agitating each solution for several hours on a rotator to infiltrate, Millonig recommends using a clinical centrifuge at 2,500 rpm for 10-15 min for each solution. Polymerize overnight at 70oC. You can section the next morning (though I prefer curing for an additional week).
This technique also eliminates the need to make multiple batches of resin or to freeze the unused resin overnight.
Feel free to contact me directly for more details.
______________________________________________________________________ Donald L. Lovett e-mail: lovett-at-trenton.edu Assoc. Professor, Dept. of Biology voice: (609) 771-2876 Trenton State College, NJ 08650-4700 fax: (609) 771-2674
} As most of you know, when Germany was reunited so was Zeiss aus Jena and } Zeiss aus Oberkochen. They are now one company. They are gradually } moving to East Germany. Nina Allen } Right.
Their address:
Carl Zeiss Jena GmbH Zeiss Gruppe Tatzendpromenade 1 a 07745 Jena Germany
Disclaimer: The views expressed in this message are those of the author, and not necessarily those of the CSIR and/or it's employees. Message-Id: {s129e944.051-at-csir.co.za} X-Mailer: Novell GroupWise 4.1
Subject: Time: 9:10 AM OFFICE MEMO More on Alcatel 220 DiffPmpOil Date: 2/20/96
In my last message I apparently gave some people the impression that Alcatel 220 diffusion pump oil is a polyphenyl ether fluid. This is not correct. It is the synthetic hydrocarbon compound eicosyl naphthalene. As shown in the graph of pump fluid characteristics on page 182, and as discussed on p 183, of the reference previously cited (I've been asked to stop mentioning this source directly, since some people think it smacks of commercialism), it has vapour pressure and boiling point characteristics very similar to the polyphenyl ether fluids Santovac-5 and Convalex-10, and is perhaps as good as these fluids in regard to thermal and oxidative stability. I believe it is also significangly less expensive than the PPE fluids. Factors to be considered in changing from one pump fluid to another are also discussed on p. 189 of said reference. W. C. Bigelow (bigelow-at-umich.edu)
The grid pattern that you said is appearing as a reflection of the SEM chamber's interior sounds like a 'charge mirror'. This usually occurs if you have a highly insulative surface which has been hit with a high energy beam to build up a charge. Then, when the beam energy is reduced to 1 or 2 KeV, the beam is repelled by the charge surface, scanning the screen of the secondary electron detector instead. Since the reflected beam continues to raster normally, you get an image of the detector screen instead of the sample. The problem is that you are viewing a metallic surface which shouldn't be charging. Is there a non-conductive coating on the ball?
One other possiblity...This is really unlikely, but I have done it before... The opening of the chamber to the diffusion pump may have a screen covering it (My JEOL 840A does). I have made the error of missing the sample altogether and imaging this screen. Sounds ridiculous, but it's true. You may want to consider this (like checking for your keys in the refrigerator).
Good Luck.
Scott D. Holt BUEHLER, LTD. 41 Waukegan Rd. Lake Bluff, IL 60044 Phone: (847)295-4546...New area code!
John- I recommend you call Bill McGee at Microtome Sevice Co (315-451-1404). Bill used to work for DuPont-Sorvall and is very experienced with parts and service for the MT-2. Steven Slap 102134,1660-at-compuserve.com
I have been persuaded (now to my regret) to give a talk about electron beam cleaning. Having posted requests for information to a number of logical sources (e.g. the surface science listserver) and only recieved information requesting that I send THEM anything useful, let me try here - at least we use electron beams! Has anyone heard of this and know any useful references or links ?
I would like to calculate the individual areas of muscle fibers in frozen muscle sections. In the past I have used a planimeter and/or a simple grid transparency. I would like to switch to a more "1990's" method, presumably using a graphics tablet and the appropriate software. Basically, the software would not only have to calculate individual fiber areas, but also what percent of the entire muscle is made up of one fiber type or another. Certainly a built in statistical package would be useful. If anyone could recommend a set-up for use with a Macintosh, I would greatly appreciate it. If there is another way besides a graphics tablet, I would certainly consider that as well.
Thank you,
Martin A. Levin Department of Biology Eastern Connecticut State University Willimantic, CT 06226 Phone: (860)465-4324 FAX: (860)465-5213
The boss is ready to send a manuscript off to a journal and has asked for the definition of High Resolution SEM vs regular good res SEM. He's looking at intramembrane particles of 10-20 nm on our Hitachi S-800 Field Emission SEM, which I lovingly always introduce as our Hi-res SEM. Are there definitions in place for hi-res SEM and TEM? I see all kinds of terms bandied about, but no numbers...
We also have a related query; Is anyone aware of any papers on non-cryo SEM of intramembrane particles? Not freeze-fracture, not frozen, hydrated, cryo SEM, and not AFM. We want to make sure to reference any related work.
Thanks and aloha, Tina
***************************************** Tina (Weatherby) Carvalho * Biological Electron Microscope Facility * University of Hawaii * (808) 956-6251 * tina-at-ahi.pbrc.hawaii.edu * http://www.pbrc.hawaii.edu/bemf/ * *****************************************
Doug Davis Staff Research Associate Electron Microscope Facility University of California Berkeley, CA 94720 (510) 642-2085 dbd1-at-uclink4.berkeley.edu
Suggest you try Ernst Keller at Zeiss in Thornwood, New York. If he can't help you directly, I'll bet he can put you in touch with those who can.
Their 800 # is 356-1090.
Good luck,
Ellie Solit The Cambrex Group
On Sun, 18 Feb 1996, Jean-Louis Beek wrote:
} Hello everyone, } } Yet again I am resorting to the listserv for advise as it has never let me down. } } I have recently purchased an old Carl Zeiss Jena compound microscope , and would like } to establish the value of it by a true connoisseur. } A brief description: } It is brass with a black base, a mirror as a light source, and a separate condenser for each } objective. A circular stage with poloriser and analyzer. } Complete in wooden box with extra eyepieces and objectives. } } Is anyone able to assist me over the net or do I have to resort to antique dealers. } Looking forward to any help I can get. } } Regards, } Jean-Louis Beek } jlbeek-at-solo.pipex.co.za } } } } } }
Dear Greg, When I need to rapid process tissues I use the following protocol:
1) Fix in 3% glut for 5 min. with a 30 sec. Microwave (MW) blast. (temp of solution not to excede 50 C) 2) Rinse 2X with sodium cacodylate for 30 sec. each. 3) Post-fix in 1% OsO4 for 15 min. including a 30 MW. 4) Rinse in s-collidine 2X for 30 sec. each. 5) Tertiary fix in saturated UA for 15 min. with a 30 sec. MW. 6) Dehydrate in ethanol (30,50,70,90,95,32X-100%) for 30 sec. each using the microwave for each step. 7) De-ethanate (like this word?) with 100% acetone 4X for 30 sec. each using the microwave. 8) Infilitrate with a 1:1 mixture of acetone:Spurr's resin (rapid cure) for 1 hour with a MW blast every 10 min. 9) Place tissue in filled BEEM capsules and place in convection oven at 70 C. 10) Every 10 min. increase temp 2.5 C and at the end of 1.5 hrs remove blocks and section. (do not excede 100 C)
Tissue should not be any bigger than 2mm, and assuming that you receive it in formalin. If not received in Formalin, increase the glut fixation to about 15 min.
In our lab any thing that is not done on the same day that we receive it, is not considered STAT. We routinely try and get samples out (at least on the scope) within 24 hrs of receiving them.
Hope this protocol works for you,
Best of luck Ed Calomeni Dept. Pathology Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
} A question for those doing thin section TEM for diagnostic pathology: } } Is there a standard protocol for rapid processing of tissues where } overnight ( or sooner) results are required? I am talking straight morpholgy.
In our lab we use the following processing procedure for those occasional rush clinical specimens: It is assumed that the specimen is delivered to the lab in 2% GA. Cut material into smaller pieces than for routine, approx. 1/2 mm3. Follow the procedure bellow:
-sodium cacodylate (0.2M pH7.4) 2x3 min 4C -osmium tetroxide (1%) in 0.1M NaCac 1x40 min 4C -70% EtOH 2x5 min RT -95% EtOH 2x5 min RT -100% EtOH 3x10 min RT -propylene oxide 3x5 min RT -50:50 Spurr's:propylene oxide 1x20 min RT -75:25 Spurr's:propylene oxide 1x20 min RT -100% Spurr's 1x30 min RT
Embed in fresh 100% Spurr's and polymerize in an oven at 85 C for 1 - 1 1/2 hours.
Sarka Lhotak EM Facility, Mc Master University Hamilton, Ontario, Canada lhotaks-at-fhs.mcmaster.ca
While I do not have the definitive answer to the beam energy/angle incidence debate, I do have several references that may be of help:
All of the following are by Dr. Arpad Barna of the Research Institute of Technical Physics of the Hungarian Academy of Sciences. If you have an interest, please contact me off-line and I can send you copies at no charge.
1) Topographic Kinetics and Practice of Low Angle Ion Beam Thinning
2) The Possibility of Surface Polishing by Ion Beam Thinning
3) Ion Beam Thinning on the Bases of Topographic Kinetics
4) Model Considerations of Ion Beam Thinning for Preparing TEM Samples
There are other papers dealing with specific sample sets such as diamond coatings and other materials that may be of interest also. There is a lot of good information in these papers but I don't recommend them for those who are fainthearted as the math can be a bit intimidating at times!
South Bay Technology does sell the IV3 Ion Mill which is produced in Hungary and is based on work done by Dr. Barna. Some proceeds, I believe, go to fund research at the Research Insitute of Technical Physics. That being said, please be aware that these are technical reports and NOT brochures on the ion mill! The theory is applicable to any ion mill. I hope this information helps!
Best regards-
David Henriks TEL: 800-728-2233 (toll-free in USA) South Bay Technology, Inc. 714-492-2600 1120 Via Callejon FAX: 714-492-1499 San Clemente, CA 92673 USA e-mail: 73531.1344-at-compuserve.com sbt-at-msa.microscopy.com
Manufacturers of Precision Sample Preparation Equipment and Supplies for Metallography, Crystallography and Electron Microscopy.
} A question for those doing thin section TEM for diagnostic pathology: } } Is there a standard protocol for rapid processing of tissues where } overnight ( or sooner) results are required? I am talking straight morpholgy.
In our lab we use the following processing procedure for those occasional rush clinical specimens: It is assumed that the specimen is delivered to the lab in 2% GA. Cut material into smaller pieces than for routine, approx. 1/2 mm3. Follow the procedure bellow:
-sodium cacodylate (0.2M pH7.4) 2x3 min 4C -osmium tetroxide (1%) in 0.1M NaCac 1x40 min 4C -70% EtOH 2x5 min RT -95% EtOH 2x5 min RT -100% EtOH 3x10 min RT -propylene oxide 3x5 min RT -50:50 Spurr's:propylene oxide 1x20 min RT -75:25 Spurr's:propylene oxide 1x20 min RT -100% Spurr's 1x30 min RT
Embed in fresh 100% Spurr's and polymerize in an oven at 85 C for 1 - 1 1/2 hours.
Sarka Lhotak EM Facility, Mc Master University Hamilton, Ontario, Canada lhotaks-at-fhs.mcmaster.ca
I have spare parts (sorry, an inventory of assorted stuff is not available) from an Hitachi HU-11E TEM that is available for anyone who wants them. Just pay for the shipping and it's yours.
Please respond to:
************************************************************************* Lucille A. Giannuzzi, Ph.D. Assistant Professor
Materials Science and Eng. Program Dept. of Mechanical and Aerospace Eng. phone (407) 823-5770 University of Central Florida fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA *************************************************************************
} I would like to calculate the individual areas of muscle fibers in frozen } muscle sections. In the past I have used a planimeter and/or a simple grid } transparency. I would like to switch to a more "1990's" method,
The infinitely valuable (& Mac Native) image processing/analysis computer program called NIH Image written by Wayne Rasband, will readily do this work for you. You will have to first digitize an image of the muscle sections ( & a variety of options are available to you here, flat bed scanners, TV cameras, slow scan CCD's) to get the data on to the Mac, but this should be easier than using the graphics tablet. This will be your limiting step, but once done then you can digitize areas & tabulate the results. By writing a simple macro you can automatically compute % or export the results to a spreadsheet program.
Using various Operating System Emulators (Executor for the PC, MAE for Unix) the program can also be run on Unix boxes & Windoze Machines. One commerical manufacturer is developing a port of this program to the PC platform.
You can download a free copy of Image (current version number is 1.60) from either the NIH ftp site (zippy.nimh.nih.gov) or the MSA ftp site (ftp.msa.microscopy.com) or the ANL AAEM WWW Site (www.amc.anl.gov) in all cases you should look for the "public" directory then filter down the directory paths until you see the imaging/data processing area.
Martin, Any respectable image analysis software should be suitable. IPLab Spectrum from Signal Analytics would be good for MacIntosh. Try NIH Image first as it is free. In any case, the difficulty will be measuring the individual fibers as segmenting the individuals from the mass via thresholding may prove difficult without clearly defined boundaries. Some image enhancement might be useful. Manual tracing might be necessary.
I can supply you with IPLab spectrum. I can supply you NIH Image with a frame grabber board for your MacIntosh. SCION supplies Image with its boards.
John Martin Levin wrote: } } I would like to calculate the individual areas of muscle fibers in frozen } muscle sections. In the past I have used a planimeter and/or a simple grid } transparency. I would like to switch to a more "1990's" method, presumably } using a graphics tablet and the appropriate software. Basically, the } software would not only have to calculate individual fiber areas, but also } what percent of the entire muscle is made up of one fiber type or another. } Certainly a built in statistical package would be useful. If anyone could } recommend a set-up for use with a Macintosh, I would greatly appreciate it. } If there is another way besides a graphics tablet, I would certainly } consider that as well. } } Thank you, } } Martin A. Levin } Department of Biology } Eastern Connecticut State University } Willimantic, CT 06226 } Phone: (860)465-4324 FAX: (860)465-5213
As far as "RUSH" processing tissue for diagnostic EM here is the method I've used for 15 years in a diagnostic EM lab. We routinely got good results and 8x10 glossies by 4 that afternoon. Hope this helps.
"RUSH" Processing for EM 3 Tissue must be no larger than 1.5mm . Well fixed.(2.5% glut in Dulbecco's PBS, pH7.3)
WASH...............PBS..................2X5 MIN. 1%OsO4 in PBS...........................20 MIN RTEMP. WASH...............PBS..................2X5 MIN *70% ETOH...............................3 MIN 95% ETOH................................3 MIN 100% ETOH...............................2X5 MIN PROPYLENE OXIDE.........................2X5 MIN 1:1 P.O.:ARALDITE 502...................30 MIN (REMOVE Caps) 1:3 " " ...................20 MIN PURE ARALDITE 502.......................10 MIN
EMBED
CURE IN 75C OVEN FOR 50 MIN. PLACE IN 95C OVEN FOR 50 MIN. PLACE IN FREEZER AND LET COOL 10 MIN.
*ALCOHOLS MUST BE FRESH.
This method worked rather well when I was at GW University. If you have any questions, call or e-mail me.
Peace,
Phil Rutledge, Director voice: (410) 455-3582 Center for Electron Microscopy e-mail: prutle1-at-gl.umbc.edu UMBC Dept. of Biology 5401 Wilkens Ave. Catonsville, MD 21228
Message-Id: {199602211734.MAA11806-at-thomas.ge.com} X-Authentication-Warning: thomas.ge.com: daemon owned process doing -bs
Does any one know an isotropic wet etch for Palladium? I am trying to make an etched pattern on Pd deposited on GaAs. The Pd is coated with photoresist and should be wet etched for making the patterns.
Thanks;
M. Dubey Research Physical Scientist Physical Science Directorate Advanced Devices Fabrication Division AMSRL-PS-DB Fort Monmouth, NJ 07703 Ph. 908-427-4040 Fx. 908-427-4306
Does anyone know of a source for an infrared microscope? I have a professor here that wants one.
Thanks
The following is an attached File item from cc:Mail. It contains information that had to be encoded to ensure successful transmission through various mail systems. To decode the file use the UUDECODE program. --------------------------------- Cut Here --------------------------------- begin 644 RFC822.TXT
A colleague of me is struggling with an immunostaining procedure.
Problem: Blocking the endogenous peroxidase in intestines from rats and mice in cryo sections. Untill now we've tried Perhydrol in different concentrations, vari‹ng between 0.3 and 2.4% with or without 0.1% sodium azide before, during and after fixation. Upto now we did not succeed blocking the endogenous peroxidase in cryo sections.
Who can help????
Winfried Leeman TNO Food & Nutrition Research Institute PO box 360 3700 AJ Zeist the Netherlands Voice: +31 30 694 44 97 Fax : +31 30 696 02 64
Does any one know an isotropic wet etch for Palladium? I am trying to make an etched pattern on Pd deposited on GaAs. The Pd is coated with photoresist and should be wet etched for making the patterns.
Thanks;
M. Dubey Research Physical Scientist Physical Science Directorate Advanced Devices Fabrication Division AMSRL-PS-DB Fort Monmouth, NJ 07703 Ph. 908-427-4040 Fx. 908-427-4306
Message-Id: {199602221434.JAA04333-at-vaxserv} X-Sender: nnicklaus-at-cave.sarnoff.com X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Depends upon your definition of "infrared". A near infrared (e.g. around 1 um) microscope may be purchased from Research Devices (NJ). Telephone 908-572-4800 and fax 908-572-4808.
Many standard microscope will also work in the near IR, just with poorer quality. In fact, Research Devices used to take standard microscope objectives, disassemble, re-space the components and reassemble.
You may adapt a standard microscope for near IR work by using a CCD camera without color correction filter at the camera port or by using an image converter. You need to check the optics of the specific microscope you have to make sure there are no selective spectral coatings, etc.
Looking in a copy of the Photonics Buyers Guide (should be in your library) yields a list of about a dozen vendors under "microscopes,infrared". Depending upon your requirements you might also call them.
----------------------------------- At 07:35 AM 2/21/96 CST, you wrote: } } Does anyone know of a source for an infrared microscope? I have a professor } here that wants one. } } Thanks } }
Hello Everyone, I need to find out what solvent can be used to dissolve polymerized LR White or Lowicryl. I am not sure which resin is the culprit. We have a Reichert-Jung CS auto cryo-substitution unit, whose specimen substitution chamber has become stuck in the substitution container (terminology taken from the owners manual). Since it has been stuck for some time, and I wasn't informed of the resin being used at the time the problem started, I can only limit it to LR White or Lowicryl. A vendor of LR White suggested that 'acrylic paint remover' might be the only hope. What common lab chemicals are in 'acrylic paint remover'. Any other suggestions? Thanks in advance,
What is the shared experience out there with HRTEM imaging using a cryogenic (LN2) sample holder? I have used the ubiquitous Gatan-type of cryogenic holder for reducing certain types of beam damage for CTEM and AEM, but now I would like to do some lattice-fringe imaging using the holder to limit beam-induced reduction of my oxide samples. How much will my point-to-point resolution be degraded by vibration, drift or other effects induced by the holder? On Gatan's Web page they seem to quote a guaranteed resolution of around 0.5 nm with 0.34 "attainable". This is not too encouraging for imaging oxide lattice spacings. I will try anyway, but perhaps I'm bound to be disappointed....Can someone predict my future?
Roy Christoffersen Texas Center for Superconductivity 3201 Cullen Houston, TX 77204-5932 roy-at-bayou.uh.edu (713) 743-8273 FAX: (713) 743-2787
Robin Griffin wrote: } Does anyone know of a source for an infrared microscope? I have a professor } here that wants one.
Do you want a system which images things, counts things, takes spectra of small regions, or all three? What sizes are the things you want to look at? In or on what kind of substrate?
These are questions to answer before I even know if I could point you in the right direction. For example, there are IR systems which can count solid state defects only 20nm in size, even providing spatial coordiants for them in three dimensions, but nonetheless form neither images (certainly not at THAT resolution!) nor spectra.
Cheers. /philf :)
//\/\/\/\---} // Phil Fraundorf Physics & Astronomy/CME 314+5165044 philf-at-newton.umsl.edu \\ B503 U.Missouri-SL St.Louis MO 63121 USA http://newton.umsl.edu/~philf \\/\/\/\/\/\/\/---}
Contrary to the suggestion that this discussion be off-line, I feel this is an area that needs greater awareness and implementation in microscopy laboratories. There are two issues here: the quality system chosen, and the method of implementation.
The ISO 9000 series is production and supply oriented, but does contains standards applicable for the laboratory. ISO/IEC Guide 25 is more laboratory oriented. In a short paper at a 1994 Paper Industry conference, one of my colleagues compared these with each other and the ANSI-ASCQ Q2 standards for laboratory quality.
Regardless of the system chosen there are many things that need doing in a microscopy laboratory to establish a quality program. They include documentation of operating procedures, regular checks of microscope and EDX performance (signal strength, resolution, magnification calibration), logging of all instrument repairs and adjustments, analytical data review, etc.. As a practical matter it has been difficult to achieve the discipline and establish priority to execute performance tests on a regular basis.
I would like to hear from others who have implemented quality programs as to their experiences.
Hi Netters... I have a question for you histology/microscopy mavens out there... we have been embedding our Golgi-stained tissueblocks in low viscosity nitrocellulose...the cut sections are cleared in alpha-terpineol and after several thorough rinses in xylene we have coverslipped the tissue using Permount (Fisher Scientific)... Recently, we have also tried using Shandon mounting mediums [Shandon-Mount and E-Z Mount] (which are methacrylate polymer-based)...we have noticed an occasional cloudiness in the rim of nitrocellulose surrounding the tissue....is there something that we can do to eliminate this??? (although it's more noticable in the nitrocellulose rim, I concerned that the cloudiness may extend into the tissue itself and might impair observations...) Thanks in advance.... Ron Mervis (RonMervis-at-aol.com) NeuroMetrix Research tel: 614-486-6080 fax: 614-486-6020
At the Marine Biological Laboratory in Woods Hole, Massachusetts, is a Postdoc position available for developing 3-D optical sectioning, deconvolution and reconstruction techniques applicable to polarized light microscopy. The techniques will be based on image data obtained with the New Pol Scope described in a recent article in J. Microscopy, Nov. 1995, Vol. 180, p. 140-147.
More information on the postdoc position can be obtained at the Biophysical Society Employment Service:
http://biosci.cbs.umn.edu/biophys/employ.html
Look under "Positions", "Postdoc", "**Marine_Bio-MA-microscopy" _____________________________________________________________ Rudolf Oldenbourg, PhD Architectural Dynamics in Living Cells Program Marine Biological Laboratory, Woods Hole, MA 02543, USA Tel:508-289-7426, Fax:508-540-6902, E-mail:rudolfo-at-mbl.edu
6th Frontiers conference on electron microscopy in materials science June 4-7th, Hyatt Regency Oak Brook Hotel, Oak Brook, IL, USA. Sponsored by ANL, LLNL, LBL, UOP, ORNL, and LANL
This conference will provide an international forum on the appliction of advanced EM techniques to problems in materials science. Plenary lectures will summarize the principles and practices of specific techniques used in the field. Additional presentations will give examples of applications.
Organizing committee: C. W. Allen ANL, W. E. King LLNL, T. E. Mitchel LANL, S. A. Bradley UOP, T. Nolan ORNL, U. Dahmen LBL, M. Wall (exhibits) LLNL
Program Committee: Magnetic materials-R. Sinclair, Quantitative-HREM U. Dahmen, Orientation imaging-H. Weiland, Advanced techniques-S. Bradley, Energy filtered imaging-J. Mayer.
Subject areas for solicited papers: Metals, HREM, XEDS, Ceramics, Electronic materials, OIM, Surfaces, EELS, Magnetic materials, CBED, Holography, Specimen preparation, etc.
For registration information contact Karen Sitzberger at LLNL, 510-423-7988
For exhibitor information contact Mark Wall at LLNL, 510-423-7162, or Mark.Wall-at-quickmail.llnl.gov (limited to 15-20 booths)
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Good day all.
I have a client who wishes to measure the volume of a liquid wetting a rough surface.
The liquid fills the "troughs" in the surface, but due to surface tension, overfills them.
Can anyone suggest a suitable technique for making this measurement?
I've been thinking along the lines of a focus calibrated scanning stage optical technique (laser confocal?), but this is by no means my area, and I'm not sure the instrument exists to do the job.
Any suggestions of optical, electron optical, or scanning probe techniques would be appreciated.
Please respond to the server, or directly to:
############################################################### # # # Don Steele STEELE-at-KRDC.INT.ALCAN.CA # # ALCAN INTERNATIONAL # # Kingston Research and Development Center # # P. O. Box 8400 # # Kingston, Ontario Canada K7L 5L9 # # # ###############################################################
Mark Darus writes:} } I'm looking at fused silica, quartz, trying to identify an inclusion } which I think is iron. I have a good K alpha iron peak, so all is well so } far, but also I have a large peak in an area where either Al or a sum peak } for the L alpha line of iron should be, but I have no peak at the area } where the L alpha should be. } My question is, can you have a sum peak, on EDX, for a particular } element, and not have a peak where the one should be that is causing the } sum peak? } I think the answer is yes, perhaps my counts are coming in too fast } and its doubling everything and showing a sum peak, but I thought I would ask.
You should also see an Fe L-alpha peak if you are seeing a sum peak for Fe L-alpha, *unless* the lower level discriminator on your EDS system is set so high as to cut off low energy lines.
If you suspect that the detector window is absorbing the Fe L-alpha lines, that detector would still absorb the two photons arriving simultaneously to create a sum peak, since each photon still only has the energy of the Fe L-alpha x-ray line.
Lowering the accelerating voltage below the critical excitation voltage for Fe K-alpha does not necessarily eliminate the possibility of a Fe L-alpha sum peak since the Fe L-alpha line will still be generated.
The best way to decide if it is a Fe L-alpha sum peak or not is to lower the x-ray count rate to reduce the incidence of two photons arriving at the same time. A simple tweaking of the beam current using a condensor lens control will do this.
(Of course an even better method would be to make the observations with a wavelength dispersive spectrometer where sum peaks are not a problem.)
Good luck,
Carl
====================================== Carl Henderson University of Michigan Electron Microbeam Analysis Laboratory 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063
In response to Randy Nessler's query on solvents for acrylic-type resins, methylene chloride works very well. This solvent can be used to completely solubilize Plexiglas and other acrylic polymers. It is a principal ingredient in commercial paint strippers.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia
Martin, The use of a point counting grid for measuring areas is certainly a 1990's approach and one advocated by the 'new stereology' group in Europe. Point counting is the most efficient use of time to get excellent estimates and takes less time than using thresholding or tracing of individual muscle fibers. If you combine NIH Image with some of the point counting macros on the NIH Image server you can capture video images of your tissue, overlay a point counting grid, score the hits within NIH image and export the data to excell. In general the maximum number of points that are needed for a good estimate is between 50 to 100. This combined with proper sampling of your tissue will yield quick and accurate results. See some of the more recent papers by Gunderson and group. You will find that this technique gives a more precise answer than the methods of planimetry or thresholding an image.
} I would like to calculate the individual areas of muscle fibers in frozen } muscle sections. In the past I have used a planimeter and/or a simple grid } transparency. I would like to switch to a more "1990's" method, presumably } using a graphics tablet and the appropriate software. Basically, the } software would not only have to calculate individual fiber areas, but also } what percent of the entire muscle is made up of one fiber type or another. } Certainly a built in statistical package would be useful. If anyone could } recommend a set-up for use with a Macintosh, I would greatly appreciate it. } If there is another way besides a graphics tablet, I would certainly } consider that as well. } } Thank you, } } Martin A. Levin } Department of Biology } Eastern Connecticut State University } Willimantic, CT 06226 } Phone: (860)465-4324 FAX: (860)465-5213
Herb Hagler, Ph.D. Director of Computer-Assisted Instruction for Southwestern Medical School UT Southwestern Medical Center (214)648-3890 Fax(214)648-3925
I read an article published many years age. It mentioned that techniques such as laser-raman spectrometry coupled with electron microscopy can offer a chance to identify molecule structures of "in situ" organic matter. Does anyone know of such kind of instrument available ? Thanks.
A fellow researcher is interested in purschasing either a SyQuest drive or a Zip drive. Are there any of you kind folks out there that have either/both. Any comments regarding either system will be much welcomed.
If this subject has been discussed before, please accept my apoligies(sp).
Thanks, Ed Calomeni Dept Pathology Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
A fellow researcher is interested in purschasing either a SyQuest drive or a Zip drive. Are there any of you kind folks out there that have either/both. Any comments regarding either system will be much welcomed.
If this subject has been discussed before, please accept my apoligies(sp).
Thanks, Ed Calomeni Dept Pathology Medical College of Ohio Toledo, OH 43699 emlab-at-opus.mco.edu
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Thanks to those who have responded to my request for technique suggestions for the measurement of a liquid wetting a rough surface.
I'm afraid "volume" was perhaps a poor choice of words on my part for the measurement I wish to make. I'm also interested in knowing how far above the nominal surface the wetting liquid is (due to surface tension), and hopefully wish to map this over a defined area. I'm expecting the cross section to be something like: ___ _/ \ liq. __/ /\ \_ / /\ / \ / substr. _/ \/ \__/
What do you think about a profilometer trace over a frozen sample, followed by a trace over the same, cleaned area?
############################################################### # # # Don Steele STEELE-at-KRDC.INT.ALCAN.CA # # ALCAN INTERNATIONAL # # Kingston Research and Development Center # # P. O. Box 8400 # # Kingston, Ontario Canada K7L 5L9 # # # ###############################################################
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please, subscribe me. thanks! Dr. Luis E. Dettin ldettin-at-cmefcm.uncor.edu Centro de Microscopia Electronica Facultad de Cs Medicas-UNC Te/Fax: 01-051-690442 CC 362 5000 Cordoba Argentina
The 100 meg Zip drive from Iomege is terrific, you can carry an enormous amount of information, including particularly greedy TIFF images, in your pocket. Kate Connolly
Hello Everyone, I need to find out what solvent can be used to dissolve polymerized LR White or Lowicryl. I am not sure which resin is the culprit. We have a Reichert-Jung CS auto cryo-substitution unit, whose specimen substitution chamber has become stuck in the substitution container (terminology taken from the owners manual). Since it has been stuck for some time, and I wasn't informed of the resin being used at the time the problem started, I can only limit it to LR White or Lowicryl. A vendor of LR White suggested that 'acrylic paint remover' might be the only hope. What common lab chemicals are in 'acrylic paint remover'. Any other suggestions? Thanks in advance,
Kovex corporation is currently developing a low-cost confocal microscope system that will provide real-time 3D images on a PC based platform. The system eliminates the use of an optical microscope in its design and is completely portable.
We are interested in receiving feedback from users and purchasers of microscope equipment in order to determine the usefulness of our products. We currently use a fixed wavelength system that reduces the flexibility of our system for the fluorescence user, but provides more opportunities for cost reduction to the user of reflectance microscopy. A complete system including software, PC, and external microscope apparatus will sell for $68,000. It is our intention to make this technology more accessible for use in quality control environments, and in industrial surface analysis. We believe our system will be used as a primary analytical tool in R&D environments where optical systems currently dominate.
Any feedback will be greatly appreciated.
Thank you,
Matthew C. Boettner President and CEO Kovex Corporation
Re} Martin Levin } I would like to calculate the individual areas of muscle fibers in frozen } muscle sections. In the past I have used a planimeter and/or a simple grid } transparency.
I agree with the reply given by Herb Hagler, the overlay counting methods are efficient, easy to use, and belong to the 1990's battery of techniques. They can be easily used on digitized images too.
However, I would like to add an additional point. If you are interested in individual fiber areas then working with the selected fibers is OK. If you are more interested in the mean fiber volume then you must take care to properly sample the whole population. Working on a few selected pictures or tracings will add a bias to your result and thus disqualify it.
To properly eliminate bias requires a defined sampling strategy which starts at the animal (or culture dish) level and continues through embedding and sectioning to photography. One paper to cover this point in a readable way is in J. Histochem. Cytochem. 40:1929-1936 1992 and is written by John Lucocq.
Best regards, Paul Webster, Ph.D. Center for Cell Imaging, Yale School of Medicine.
Mark, You do not say if you have a detector capable of detecting low energy=20 ( {1keV) x-rays. If you do not then the peak cannot be an Fe L sum peak as=20 the Fe L x-ray will not reach the detector crystal and therefore cannot the= n=20 cannot create signals which can contribute to the sum peak . Eric ----------
{ {File Attachment: UUCP_ENV.TXT} } Mark Darus writes:} } I'm looking at fused silica, quartz, trying to identify an=20 inclusion } which I think is iron. I have a good K alpha iron peak, so all is well so } far, but also I have a large peak in an area where either Al or a sum peak } for the L alpha line of iron should be, but I have no peak at the area } where the L alpha should be. } My question is, can you have a sum peak, on EDX, for a particular } element, and not have a peak where the one should be that is causing the } sum peak? } I think the answer is yes, perhaps my counts are coming in too fas= t } and its doubling everything and showing a sum peak, but I thought I would ask.
You should also see an Fe L-alpha peak if you are seeing a sum peak for Fe L-alpha, *unless* the lower level discriminator on your EDS system is set so high as to cut off low energy lines.
If you suspect that the detector window is absorbing the Fe L-alpha lines, that detector would still absorb the two photons arriving simultaneously to create a sum peak, since each photon still only has the energy of the Fe L-alpha x-ray line.
Lowering the accelerating voltage below the critical excitation voltage for Fe K-alpha does not necessarily eliminate the possibility of a Fe L-alpha sum peak since the Fe L-alpha line will still be generated.
The best way to decide if it is a Fe L-alpha sum peak or not is to lower the x-ray count rate to reduce the incidence of two photons arriving at the same time. A simple tweaking of the beam current using a condensor lens control will do this.
(Of course an even better method would be to make the observations with a wavelength dispersive spectrometer where sum peaks are not a problem.)
Good luck,
Carl
ffffffffffff=3DCarl Henderson University of Michigan Electron Microbeam Analysis Laboratory 2501 C.C. Little Bldg. Ann Arbor, MI 48109-1063
I am trying to locate a Leica microscope dealer for info on a slide-in filter to remove the endogenous fluorescence produced by chloroplasts when using a Rhodamine filter-pak.
Anyone have any info on the characteristics of such a filter? On another brand of microscope, such a filter is numbered 575DF40 and 9336. I assume the 575 refers to the wavelength allowed to pass.
Thank you.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Immediate Opening for SEM Analysis Engineer at SVC:
Principal Duties & Responsibilities:
SEM analysis of display components; basic failure analysis. Responsible for upkeep of JSM 820, with full JEOL service contract; Supervision & training of other SEM users.
Special work circumstances:
It is preferred that candidate be able to work either an early (start -at- 6AM) or late (finish -at- 8PM) shift.
Experience & Education Requirements:
B.S. in Physical Science or Engineering or equivalent. Minimum 3 yrs. experience in Electron Microscopy.
Company Background:
In just 4 years Silicon Video Corp. has become one of the hottest start ups in Silicon Valley. We are developing a new class of flat panel display - Thin CRT. Millions in funding from the government have been obtained. Large corporate partners in industry and academia connections have been secured. We're searching for highly qualified candidates to join our team.
QUESTION: } } A fellow researcher is interested in purschasing either a SyQuest drive or a } Zip drive. Are there any of you kind folks out there that have either/both. } Any comments regarding either system will be much welcomed. } } If this subject has been discussed before, please accept my apoligies(sp). } } Ed Calomeni
RESPONSE:
I use both. I have far more experience with SyQuest. No problems with SQ ever. I tend to favor SyQuest because the head is never in contact with the platter which is rigid and substantial. SyQuest has a tremendously larger user base and they are the standard in the publishing industry. There are presently more of the larger (5.25") SQ platters than the 3.5" SQ's. ZIP drives use a floppy medium which is held away from the heads by air flow. Speed wise they are both similar. Replacement platters (disks) are considerably cheaper with ZIP drives - about half. So, it dependends on whether price is an issue or if ready exchange of disks is the concern. ZIP's are new and so we don't know yet if they will be as reliable as SQ's. Costs for the drives are similar ($200 for the 100 MG drives). You know, the ZIP folks, IOMEGA, have now come out with the JAZ drive, a 1 GB disk that is very cheap and the drive costs $400-500.
############################################################################# John J. Bozzola, Ph.D., Director Center for Electron Microscopy Southern Illinois University Carbondale, IL 62901-4402 U.S.A. Phone: 618-453-3730 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/departments/shops/cem.html #############################################################################
Postdoctoral Position Available at the University of Chicago
in the following areas of research
} RED CELL CYTOSKELETON
} Red cells are able to elastically deform. The extraordinary } importance of this property can be appreciated from recalling that, } although the human red cell is a biconcave disk eight microns in } diameter, it can easily pass through capillaries half this size. } This remarkable feat is accomplished by the red cell transiently changing } its shape during passage after which its well-known biconcave } form spontaneously and nearly instantly recovers. We are studying } the structural basis of this property using cryoelectron microscopy. } } McGough, A. and Josephs, R. (1990) On the structure of } erythrocyte spectrin in partially expanded skeletons. Proc. Notl. } Acad. Sci. USA 87: 5208-5212. } } Li Li and Robert Josephs. Cryo-Electron Microscopy of Human } Erythrocyte Membrane Skeletons. Proceedings of the 51st annual } meeting of the Microscopy Society of America San Francisco } Press, San Francisco. p. 116 (1993). } } SICKLE CELL HEMOGLOBIN
} Sickle cell anemia is caused by the intracellular polymerization of } sickle cell hemoglobin to form rod-like fibers. A knowledge of the } fiber structure could be used for the design of an agent that could } block fiber formation. We have combined the structure of hemoglobin } (known from X-ray crystallography) and the molecular coordinates } of the fiber (determined from electron microscopy) in order to } synthesize a model which shows the intermolecular contacts of the fiber. } This approach has allowed us to determine the contacts which form } between molecules in the fiber. We are now studying how various mutations } (some obtained by site directed mutagenesis) affect fiber structure. } Such studies are expected to account for the structural and chemical } properties of fibers in terms of intermolecular interactions. } } Watowich, S., Gross L., and Josephs, R. (1989) Intermolecular } Contacts within Sickle Hemoglobin Fibers. J. Mol. Biol. 209: 821-828. } } M. R. Lewis, L. J. Gross, and R. Josephs. Variable Pitch in Frozen } Hydrated Sickle Hemoglobin Fibers: An Image Analysis Model } Study. Ultramicroscopy, 56 303-317 (1994). } } Michael R. Lewis, Leon J. Gross, and Robert Josephs. Cryo- } Electron Microscopy ofDeoxy-Sickle Hemoglobin Fibers. } Microscopy Research and Technique 27 459 - 467 (1994). } } } ACETYLCHOLINE RECEPTOR } } The acetylcholine receptor is responsible for transduction of the nerve } impulse to muscular contraction. The receptor lies at the neuromuscular } junction in the muscle membrane. It has five subunits denoted as } a1,a2,b,d,g. Current work in our lab involves labeling different } subunits with monoclonal antibodies against defined defined sequences } in order to determine the subunit arrangement (which appears to } still be a matter of debate in spite of the considerable } work done in other labs). We are using two dimensional crystalline } tubes in order to determine the location of the labels.
} Interested individuals please contact
} E-mail: Bob-at-befvax.Uchicago.Edu } } } Robert Josephs } The University of Chicago } 920 East 58th Street } Chicago, IL 60637 }
I am by no means an expert in this field but I understand there is a process known as Electron Beam energy filtering. You might try this as it could take out all the nasty bits and may clean the electron beam.Patrick Echlin Cambridge UKOn Tue, 20 Feb 1996, L.D.Marks wrote:
} I have been persuaded (now to my regret) to give a talk about } electron beam cleaning. Having posted requests for information to a number } of logical sources (e.g. the surface science listserver) and only recieved } information requesting that I send THEM anything useful, let me try here - } at least we use electron beams! Has anyone heard of this and know any } useful references or links ? }
EmLab wrote: } } Hi all, } } A fellow researcher is interested in purschasing either a SyQuest drive or a } Zip drive. Are there any of you kind folks out there that have either/both. } Any comments regarding either system will be much welcomed. } } If this subject has been discussed before, please accept my apoligies(sp). } } Thanks, } Ed Calomeni } Dept Pathology } Medical College of Ohio } Toledo, OH 43699 } emlab-at-opus.mco.edu
Syquest drives are fine, but media will cost about the same as for rewritable magneto-optical media ($200 / cartridge). Syquest, of course, is comparatively lower priced than magneto-optical. However, for serious image storage, Pinnacle Micro's 4.6 GByte M/O drive is great! At around $1700 it is more than a Syquest, but when one considers the cost of media, may actually be quite economical. I will send you additional information via mail.
EmLab wrote: } } Hi all, } } A fellow researcher is interested in purschasing either a SyQuest drive or a } Zip drive. Are there any of you kind folks out there that have either/both. } Any comments regarding either system will be much welcomed. } } If this subject has been discussed before, please accept my apoligies(sp). } } Thanks, } Ed Calomeni } Dept Pathology } Medical College of Ohio } Toledo, OH 43699 } emlab-at-opus.mco.edu
Regarding my last message about Pinnacle Micro magneto-optical drives, you can check out Pinnacle at http://www.pinnaclemicro.com.
Dear Colleagues, I am using plastic-embedded (Lowicryl or JB-4) sections of fetal kidneys and would like to use standard avidin-biotin-peroxidase techniques for immunohistochemistry. To date, we are unable to achieve specific staining. We have tried two different etching protocols and a few of the JB-4 (but none of the Lowicryl) sections survive, but still no staining. Most references give little information regarding incubation times, etc. Can anyone help me trouble-shoot or point me towards a good reference? Thanks in advance. Victoria F. Norwood, M.D. Department of Pediatrics University of Virginia vfn6t-at-dmt03.mcc.virginia.edu
The Call for Papers and Instructions for Manuscript Submission is now (~90%) on-line on the MSA WWW site.
You may download/access the information at:
http://www.msa.microscopy.com
Look for the hotlink called "Registration & Meeting Info."
There are a few forms that still need processing, however, they will be on-line in a few more days.
If you are still waiting for your HardCopy of the Call for Papers this information will allow you to get started on your Manuscript preparation. All the information in the HardCopy Booklet (except for some artwork) will be reproduced here.
Several new options have been also added to the MSA WWW Site.
1.) New Search Engine Access
A new options for direct connection to WWW Search Engines has been implemented. Just enter your search criteria and choose your favorite search engine.
2.) New Aliases to Society Office Holders
You may directly Email the office holders of MSA by using the aliases shown below. Direct links are also provided on the MSA WWW Page.
President (MSAPresident-at-MSA.Microscopy.Com) includes Past-President & President-Elect
Secretary (MSASecretary-at-MSA.Microscopy.Com)
Treasurer (MSATreasurer-at-MSA.Microscopy.Com)
Council (MSACouncil-at-MSA.Microscopy.Com) reaches all current Council Members
Business Office (BusinessOffice-at-MSA.Microscopy.Com) direct to the Business Office
WebMaster (I'll let you figure out this one)
These links are aliases and provide you with a method to always reach the current office holder.
Ed, I have the SyQuest EZ135 at home. I've had no problems with it at all. Access time is fast (the manufacturer claims it is significantly faster than the Zip drive). As a matter of fact, its so fast that I use it as a working hard drive. The 135 Mb cartridges are about $23.00 each (less if you buy in bulk). It is also quite small; about the size of an external CD ROM drive. I have never used a ZIP drive, so I can't make any real comparisons. I can only assure you that the EZ135 is a great little drive.
Marty
PS I don't work for or have stock in SyQuest.
} Hi all, } } A fellow researcher is interested in purschasing either a SyQuest drive or a } Zip drive. Are there any of you kind folks out there that have either/both. } Any comments regarding either system will be much welcomed. } } If this subject has been discussed before, please accept my apoligies(sp). } } Thanks, } Ed Calomeni } Dept Pathology } Medical College of Ohio } Toledo, OH 43699 } emlab-at-opus.mco.edu
Martin A. Levin Department of Biology Eastern Connecticut State University Willimantic, CT 06226 Phone: (860)465-4324 FAX: (860)465-5213
From Bob Citron: } Hi Randy; } } I would suggest using methylene chloride. This works best for our PMMA } (acrylic)lens products. } We have the same problem. The trouble with suggesting solvents like MeChloride, Chloroform, or other such things is that the R.-J. cryo-substitution unit is a good-sized piece of floor-occuping equipment, and can't be put in a fume hood. I wouldn't particularly want to be in a lab full of (e.g.) MeChloride fumes; we're exposed to enough toxins and carcinogens as it is. Any less noxious ideas? Phil Oshel
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
Regarding which removable storage drive to choose: be advised that Iomega Zips are outselling EZs all across the country by a large margin, and also that every EZ drive sold by Syquest *loses* a significant sum ($50-$100) for the company. Syquest recently announced such poor earnings (a loss of about $3.00 per share) that they have *laid off 60% of their world wide work force*. The media used for the Syquest cartridges is defective 270MB disks (one side is used) and has *no chance* of becoming the next "standard" which replaces the 1.4MB floppy, while there is an excellent chance that the Zip 100MB disk *will* become the new standard. It is expected by many that Syquest cannot continue to bleed money if they are to remain a viable company, and may have to abandon the EZ as a competitive product.
Our laboratory has adopted Zips almost universally (I know of only 1 EZ locally), and to date all of our external users are following suit. Among a 7 member group, we have 20 or more Zips either in the lab, our offices, or at home. Check your local retailers (Circuit City, CompUSA, etc.) and see what is selling. I think you will find that if you purchase a Syquest EZ, you will become a member of a small minority, and will run the risk that you have an obsolete drive that uses media which will not be compatible with the future standard (whatever it may be). Of course, you only risk a couple of hundred bucks, and 20 bucks per cartridge, so it is not a large risk.....
good luck.
Larry PS as I have posted here before, I *do* own (and continue to recommend) Iomega stock, so do your own homework.... ;-)
} Dear Victoria, Many antigens can be stained in JB-4 or Lowicryl without etching of any kind. Many etching procedures are bad for antigenicity, so you might try seeing what you get without etching at all.
And, you did not say if you were amining for light microscopy or em. But if you are working in light microscopy and you find that you are unable to get good staining from Lowicryl or JB-4, you might try the removable methacrylate system. This is a mixture of methacryates without crosslinker, so you can remove the resin nearly completely with a 10 min incubation in acetone. We have had very nice results with this for immunocytochemistry at the light level. If you are interested, I can provide more details.
Cheers, Tobias
} I am using plastic-embedded (Lowicryl or JB-4) sections of fetal kidneys } and would like to use standard avidin-biotin-peroxidase techniques for } immunohistochemistry. To date, we are unable to achieve specific staining. } We have tried two different etching protocols and a few of the JB-4 (but none } of the Lowicryl) sections survive, but still no staining. Most references } give little information regarding incubation times, etc. Can anyone help me } trouble-shoot or point me towards a good reference? Thanks in advance.
Allow me to add my voice to the chorus for Zip drives. I use one at home, and it works well. Two points I haven't seen mentioned: the 100MB discs aren't much larger (mostly thicker) than regular floppies, but are rugged, and the drive itself is small enough to take with you if you travel. Take the Zip Tools & Install discs with the drive & you can use your Zip anywhere others have the same computer as you (Mac or DOS). Phil Oshel
&&& Illigitimi non carborundum &&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&&& Philip Oshel Center for Electron Microscopy University of Illinois (217)244-3145 oshel-at-ux1.cso.uiuc.edu
I'm looking for titres of books on BIOLOGY LABORATORY TECHNIQUES. I opened a site for amateur scientists www.best.com/~funsci and I need information about experimental techniques in biology, microscopy, etc... suitables for scool or research applications. Can you help me? Please, send the answers here: funsci-at-best.com Thanks George
In the discussion of removable media I have not read anything about "storage life" of data. Does anyone know about predicted life for data/images stored on Syquest, Zip, CD, or magnetic tape cartridges? Will all of these media require re-copying every five years to ensure data integrity? That also begs the question as whether of any of these drives will around in five years to read them.
Can't answer your question exactly. We have an internal Syquest drive and it is rather old (3-years). We have come to question its reliability because two of the drives had to be reformatted after about six months of use.
I have been pleased with the IOMEGA products. We have some older Bernoulli drives around. One of the five drives we have apparently developed a sticky mechanism after about 5 years, but otherwise has been good.
} A fellow researcher is interested in purschasing either a SyQuest drive or a } Zip drive. Are there any of you kind folks out there that have either/both. } Any comments regarding either system will be much welcomed. } } If this subject has been discussed before, please accept my apoligies(sp). } } Thanks, } Ed Calomeni ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
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From my reading it seems that magnetic media has an archival lifespan of about 10 years whereas data archived on CD-ROM has an archival lifespan of 30 years. Since I plan on retiring in 25 years, my bet is to archive on CD-ROM and not have to repeat the archival process every 8-10 years.
The upside of magnetic media is the speed at which you can archive I guess, the latest CD-R drives appear to need about 40minutes to "burn" the data on disk.
Kevin McCarthy
Regarding lifespan of magnetic media, David Rothbard wrote:
In the discussion of removable media I have not read anything about "storage life" of data. Does anyone know about predicted life for data/images stored on Syquest, Zip, CD, or magnetic tape cartridges? Will all of these media require re-copying every five years to ensure data integrity? That also begs the question as whether of any of these drives will around in five years to read them.
David Rothbard
-- Institute of Paper Science and Technology
Kevin McCarthy Assistant Professor Department of Cell Biology Digital Imaging Microscopy Facility University of Alabama at Birmingham Birmingham, Alabama 35294 Phone 205-934-9923/9924 Fax 205-934-7029 "Seeing the World Through Different Eyes"
FYI: The MSA Technologists' Forum is sponsoring a Roundtable discussion at this year's Microscopy & Microanalysis meeting in Minneapolis. The theme of the roundtable is "Coping with Regulations: ISO and OSHA". As the session chair, I'm glad to see this thread on the Listserver. I encourage everyone with an interest in how regulations govern our lives in microscopy labs to attend this session. Further details will be available prior to the meeting.
Regards, Bev Maleeff SmithKline Beecham Pharmaceuticals Toxicology-US, UE 0462 709 Swedeland Road King of Prussia, PA 19406 610/270-7987 610/270-7202 fax Beverly_E_Maleeff-at-sbphrd.com ***note the new e-mail address***
Don Chernoff writes "Could someone explain how the overlay counting methods can give areas of individual fibers?"
Cross-lattice overlays (see Weibel, E. R. 1979. Steeological methods, vol 1. Practical methods for biological morphometry. Academic Press, New York.) are transparent sheets which have grids (or lines) of known dimensions on them. A simple overlay would be a square grid. For area counts, the corner of the grid is taken to represent the whole square. Therefore if the points over the structure to be sampled are counted this is the same as counting the number of squares. On samples of properly randomized specimens, this is a very accurate way of estimating area.
Since the sections that are sampled have a thickness, the area can also be used to estimate volumes.
The test lines can also be used to accurately estimate length and thus surface area.
Only small numbers of micrographs are required to do these analyses, counting (unlike tracing) does not need to be accurate, and estimations can be rapidly performed.
Mr-Received: by mta PETVAX.MUAS; Relayed; Mon, 26 Feb 1996 12:29:11 -0400 Mr-Received: by mta PETVAX; Relayed; Mon, 26 Feb 1996 12:29:11 -0400 Mr-Received: by mta SRVR01; Relayed; Mon, 26 Feb 1996 12:37:14 -0400 Disclose-Recipients: prohibited MSA Microscopy Mailing List {MICROSCOPY-at-Sparc5.Microscopy.Com} Message-Id: {1011291226021996/A03485/PETVAX/11A2D31D0A00*-at-MHS} X-Envelope-To: David.Rothbard-at-ipst.edu, MICROSCOPY-at-MSA.Microscopy.com Autoforwarded: false Mime-Version: 1.0 Content-Type: TEXT/PLAIN; CHARSET=US-ASCII Content-Transfer-Encoding: 7BIT Importance: normal Priority: normal Sensitivity: Company-Confidential Ua-Content-Id: 11A2D31D0A00 X400-Mts-Identifier: [;1011291226021996/A03485/PETVAX] Hop-Count: 2
All interested persons in removable, mass storage medias should check out the March issue of PC World. It includes a very comprehensive article on this very topic including speed, storage capacity, durability, archival quality, etc.
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All interested persons in removable, mass storage medias should check out the March issue of PC World. It includes a very comprehensive article on this very topic including speed, storage capacity, durability, archival quality, etc.
Hello I need information about,condition of analysis , about voltage,current,time deat, size of beam in Quantitative analysis and calibration For analysis , in alluvial-gold (free gold). For analysis on silicates . Anybodi are using X-PHI software ? i have problem, with K alpha line and "K" kalium . My name is Alfonso Goldschmidt i am working in the University of Chile in the GEOLOGICAL DEPARTAMENT. I am working with the old microprobe CAMECA 46 and i am try of restart the use, of this machine.
I have been asked to do TEM and/or SEM on Hepatitis C virus protease. I have a biology background that deals with tissues not crystals. They would like to examine the crystal structure of this protease. I understand that it does not form a pellet and does not separate out no matter how hard you spin it. The concentration of the sample is approximately 1 mg/ml. Where should I begin ???? Could I do an air dried sample for scanning ??
Thank You,
Barbara Hartman Schering-Plough Research E-Mail: Barbara.Hartman-at-Schering-Plough.sprint.com Phone: 201-579-4343 Fax: 201-579-4211
I agree that magnetic media has a lifetime of 10 years or so, whereas optical media is far longer, say 30 years. However, I don't believe I have *ever* accessed original data (say negatives) for any useful purpose even 5 years after original recording. Magnetic media has an overwhelming advantage in daily usage, because of its speed, portability (the frisbee test), and competitive price of media and drives. However, for the convenience of our users, in our lab we record archival data (TEM images etc) at present on CD-ROMs, and provide either Zips or CDs to our users to carry their data home. But I certainly do not worry about how I will read that archived data 25-30 years from now, because there is no question that many generations of drive devices will supplant anything that is currently available. We no longer use enlargers to print our negatives, but have scanners that handle transparencies to use if we want to make some new prints. When the time comes, we will transfer our archived images and other data to the next generation of storage devices.
BTW, recently I installed a 400K diskette containing data from May 1984 recorded from my old 128K Macintosh. My Powerbook Duo288c running MacWrite II opened a MacWrite file with no problem, and a recent version of MacPaint opened an old MacPaint file. Of course, I had absolutly *no* interest in any of that data after nearly 12 years.
Greetings, I mentioned a removable methacylate system for immunocytochemistry and several netters have asked me for more information. For this reason, I am posting the reply publicly, in case others are interested as well.
The basic resin is simply a mixture of butyl and methyl methacrylate (80% butyl). This was a resin much used in the em *before* the invention of epoxies, and now generally abandoned. But with the upsurge of immuno techniques at the light microscope level, several groups in the late 80's ressurected this simple resin system because it has the advantage of being easily removed after sectioning by a brief rinse in an organic solvent, such as acetone. Removablility is an advantage because you gain great access for your antibody to your antigen. In this respect, the removable methacylate system is like paraffin embedding; only with methacrylate, the structural preservation of the sample is generally much better than with wax. So, if you are going after sub-cellular detail, then methacrylate could be the resin for you.
If this sounds interesting, here are some references:
Baskin et al 1992 Planta 187:405 - 403. Gubler 1989 Cell Bio Int Rep 13:137-145. Van de Kant et al 1988 Histochem J. 20, 335 - 340. (animal tissue).
A protocol where protein localizations and in situ RNA localizations are done on alternate sections: Kronenburger et al. 1993 Cell Biol Intern. 17: 1013-1021.
A few notes. We drive polymerization of the methacrylate with UV light (a catalyst is added to the resin), and this is a free radical based mechanism. We were only able to get this method to work reliably with plant material by including 10 mM DTT in the resin. This seems to prevent radicals from attacking the tissue but not to prevent them from doing their polymerization thing. Also, polymerization is inhibited by oxygen. In the reference to my own work above, polymerization was done in flat molds in a box exposed to a flow of nitrogen gas. We now do embedding in capsules, under normal air with no problems. Also, as far as practical details go, compared to what we published in the Planta paper above, we have found that there is no need to put DTT in the ethanol series for dehydration, and we can compress the entire dehydration infiltration and embedding schedule to two days, rather than six.
I hope the above helps. If anyone has questions about specific steps or other details, please feel free to contact me.
! ! ! ! ! ! ! ! BACK BY POPULAR DEMAND ! ! ! ! ! ! ! !
The Second Annual Symposium on
"Integrated Microscopy"
on
September 20 - 22, 1996
at
The Wisconsin Center 702 Langdon Street Madison, WI
Program details will be posted at a later date. Information is also available on our World Wide Web page:
http://www.bocklabs.wisc.edu/imr/imr.html
#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#* Presentations will focus on biological problems for which a combination of microscopies [i.e. integrated microscopy] has been used. The speakers will demonstrate by example the power, potential and limitations of various microscopical techniques. The techniques which will be discussed include: DIC, Confocal, Multi-Photon Excitation Imaging, SEM, TEM, Cryo specimen Preparation. #*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*#*
******************************************************************************** Following the symposium, the IMR will be conducting a 2-day workshop (Sept. 23 and 24). We will be presenting lectures and provide "hands-on" experience for the following techniques: * 2-photon excitation imaging * 4D DIC imaging * Cryo-SEM * High pressure freezing * Reversible embeddment for SEM and TEM
Workshop attendence will be limited to 20 participants.
In the analytical facility at Caltech we have used both the Syquest 44 Mb and the Zip 100 Mb removable-cartridge units. I have also used a Syquest 270 Mb unit for a few days. My selection of these units has been based on performance, speed, reliability, and cost of the media for outside users.
My observations (all these systems were used on primarily on Macintosh systems, except as noted):
1. The Syquest 44 became an early standard among the Macintosh crowd, many people had them and they were used to transfer all sorts of software, data, digital images, etc. The MAS computer workshop sort of settled on this format for about 2-4 years, and we could count on being able to transfer things with our collegues. The units were nearly bulletproof with a few exceptions. Cartridges formatted on your unit at home could potentially have problems on another drive if the head alignment differed drastically. I observed this fewer than 3 times in the 5 years I have used these units. As far as reliability, I have two units that were on continuously for about 5 years with no problems. The units are loud and noisy, but have speeds rivaling older hard drives. The 44 Mb format is essentially history now, although many have $$ invested in cartridges (I have 45 of them...being consolidated onto Zip 100's now). There have been other formats available (88Mb, 105Mb, 200Mb, 270Mb), and I think the 270 Mb format is the only one that has achieved any kind of "standard" status.
Which reminds me to comment on the "longevity of media xyz". I think that currently the technology is evolving more rapidly than the useful life of any given media. For example, I have some Apple II disks with some important stuff on them, but there are no Apple II's around here...so even though the media is still (probably) ok, I don't have access to the devices used to read/write this information. Same goes for all the DEC floppy disks, RL02 platters, TK50 tapes...etc. The bottom line is that we now have the obligation to periodically transfer *all* our important archived data to the current "standard" which (probably) won't be in use 10 years from now. The question "will the data on a floppy be readable 25 years from now" is just plain academic, because those floppy drives will be found only in the Smithsonian. The worst part is that by the time you *realize* that you have archaic media, you find that it is obsolete in the sense that you cannot find a system to read it on. And as the capacity of media ever increases, so does the possiblity of losing really large sets of data. But this is a discussion best held over a few beers, I guess :)
2. The Syquest 270 is nice, but I found the unit to be very noisy, and the ejection lever felt as if it was going to break off every time I used it. The quality level is not what I would expect (but bear in mind this is a S 270 mechanism in some vendors box, so the packaging quality and noise level may vary by manufacturer). I had an insurmountable level of difficulty trying to format and *mount* PC-format 270 disks on my Powermac 8100. There are a number of drivers out there, all advertising that they mount everything, but few of them actually do the job. In our facility we have mostly Mac users, but some PC users, so we must be able to provide both formats.
3. I have had a Zip 100 Mb unit for about 9 months. It is quiet, fast and almost trouble free (a pre-formatted cartridge refused to mount and had to be reformatted; I have not had this problem with any other disks). I observed roughly similar levels of performance for a Zip 100 on a 486 PC (parallel port) and the SCSI version on a Powermac 8100; they both rival hard drive speeds. Using the Iomega driver software on the Mac, the disks can be either formatted for the Mac, or by using the 'erase disk' menu on the Mac, a disk can be PC formatted, so this unit essentially solves the Mac-PC format problems that the Syquest units apparently have more difficulty with (I'm not an expert here, just giving you my experience).
The Zip unit is nice because it has auto mount and auto eject features. Put the disk in the unit and it appears on the Mac desktop; drag it to the trash and it dismounts and ejects. The Syquest units require fooling around with that eject lever, and novices nearly always seem to want to eject the cartridge before it is dismounted (and therefore spun down -- a good way to damage the disk). This is a matter of convenience, but it adds up over the years in cumulative hassle factor.
There are caveats with the Zip 100 unit. There is no power on/off switch, just a 12 volt converter plug and line cord, so the unit must be left on all the time unless you set up a power strip and use that switch to turn the thing on and off. The unit has only SCSI device numbers 6 and 7 available, which may cause ID conflict problems on some systems, but has not been a problem with our setup.
Perhaps one of the biggest limitations is for those who plan to use the Zip 100 for backup rather than just external storage. My understanding is that you can only backup 100 Meg maximum with the Iomega backup software (i.e. 100 Meg maximum as a contiguous save set), so you are out of luck if you have a larger hard drive (which almost everyone has now). So you would have to do a backup manually.
4. The Iomega Jaz drive (about $500-600 for the drive, 1 Gig cartridge about $90) sounds like the best solution for our facility, where users can amass lots of digital images and analytical data. I have not seen one in action.
5. I just started seeing ads for another Iomega cartridge drive on tv (not the Zip, and not the Jaz, and I forget the name). Does anyone know about this?
From my experience base, I would recommend the Zip 100 for the super cheap external storage solution -- when it becomes history, we will not have invested big bucks and can simply copy to a new system and move on. The Jaz drive sounds good for those with bigger storage needs, and I anticipate getting one if there are no bad reviews. I was a long time Syquest fan, but I'm not convinced that they are in for the long haul anymore, and have been really sluggish to respond to the Iomega market pressure. Syquest has no competition for the Jaz drive that I know of...
Remember that one of my main priorities is to offer cheap high capacity media to users of the facility, some of whom come to visit only once and need to take all their data away with them. More expensive fancy optical disks etc. are not reasonable to ask these people to purchase, because they have to go read all this data back at their lab on an equivalent system. So my decision for external storage automatically commits others to purchasing the same hardware. A cd writer would be great, but these are still expensive relative to the cheap Zip solution (for example).
My opinions only, and I have not the slightest financial interest in any of this, except as a consumer. I'd rather go flying than pay big money for storage media!
Paul Carpenter
+------------------------------------------------------------+ | Paul K. Carpenter | | Division Analytical Facility | | Geological and Planetary Sciences MC 170-25 | | California Institute of Technology | | Pasadena, CA 91125 | | 818-395-6126 (X-ray Lab) 818-568-0935 (FAX, Departmental) | | paulc-at-arms.gps.caltech.edu | +------------------------------------------------------------+
Paul Carpenter:} } I just started seeing ads for another Iomega cartridge drive on tv (not the Zip, and not the Jaz, and I forget the name). Does anyone know about this? { {
Paul:
I'm jealous; you have probably seen the new TV ad blitz by Iomega which highlights the Ditto Easy 800 tape drives. These drives offer 800 MB for about $149, and have elegant software which continually backs up your work even while you are on-line, as I understand. Since Iomega can make plenty of these, the supply pipeline is easily kept filled, and even though they are the hottest selling tape backup these days, Iomega has initiated an advertising campaign to establish their name (sort of like Intel Inside) in the data storage area, prior to future advertising for Zip and Jaz when they become more plentiful. I haven't used the Ditto drive as yet, but plan to purchase one for each of my 3 Macs at home. The Zip travels around with me at present. At the lab, we have networked tape backup which is done automatically every night. I think the drives are Colorado units, which are currently being outsold by the new Dittos. There is also a Ditto Easy 3200 which offers 3.2MB for about $450, for those with more formidable backup needs.
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
Bob Craig wrote:
} The first question that one must ask is why do I need registration, } certification or accreditation? The answer to this question is usually } market driven, at least it was in our situation.
Bob is quite right about everything that he said, however there are still more reasons why one might want to get accredited.
Let me give some additional reasons:
1] As an independent laboratory, our big fear is professional liability risk. It is, I am convinced a generally under appreciated risk, but anyone offering professional opinions and advice, including microscopy, takes on a certain greater-than-zero risk. Sometimes the risk can be considerable especially if you have an unhappy client. I have found in our own analytical laboratories that having the A2LA Iso Guide 25 accreditation in place is the cornerstone of the very best loss prevention program imaginable. It sets the standard for quality and excellence, and there are outside inspectors who come in once every two years to make sure that all procedures are being followed.
Now this will not guarantee that one won't be sued, but the program surely does eliminate a good many of the weak spots that exist in the typical microscopy laboratory operation. Although I might beg, cajole, encourage, talk nice, talk harsh to certain individuals in our laboratories, nothing has more impact than having that outside assessor say the same thing as part of an audit!
2] For those in the laboratory analytical (services) business, from an insurance and underwriting standpoint, we have found that to whatever degree we incur costs associated with the accreditation, at least some of those costs come back to us by way of reductions in the premiums paid for our professional liability insurance.
3] Furthermore, since the A2LA accreditation and adherence to ISO Guide 25 is entirely consistent with any good TQM process, to the degree that the accreditation makes our TQM approach that much more effective, our cost of rework goes down.
There is a bottom line here that might come as a surprise: At least in our case, there does not seem to be any "net" cost to become accredited. Whatever money is spent to become accredited, and to fill out the extra paperwork, we believe it comes back to us "with interest" by way of increased business (from clients where quality really counts), lower insurance costs, and a smaller number of samples that have to be done over again because they weren't done right the first time.
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
Does any one knows any reference about Freeze Drying Technique, for preparation of sections for LM ?
Thanks
/-------------------------------------------------------------------------\ | Rui Costa | e-mail: ruicosta-at-esb.ucp.pt | | Escola Superior de Biotecnologia |telephone: 351-2-5580044 | | Rua Dr Antonio Bernardino de Almeida |fax: 351-2-590351 | | 4200 PORTO | | | PORTUGAL | | \-------------------------------------------------------------------------/
David Rothbard wrote: } } In the discussion of removable media I have not read anything about } "storage life" of data. Does anyone know about predicted life for } data/images stored on Syquest, Zip, CD, or magnetic tape cartridges? Will } all of these media require re-copying every five years to ensure data } integrity? That also begs the question as whether of any of these drives } will around in five years to read them. } } David Rothbard } } -- } Institute of Paper Science and Technology
Pinnacle Micro claims 30 years for its magneto-optical storage media.
I want to install a SyQuest drive (currently on a Mac) on a PC. Who, other than SyQuest, sells an easy to use software driver package for that purpose? Is any freeware available?
Thanks.
Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
Hello Pacqui, we are a commercial lab doing TEM of semiconductors. We still use 'old' techniques, i.e. we don't use Ron Anderson's tripod polisher and 3M diamond impregnated plastic polishing films. This is partly due to inertia, and partly because of cost; we can make up to 10 cross-sections per day per person just using a few glass slides and two polishing wheels. It would be nice to upgrade to the tripod technique, but at the moment we can get by with the cheap and cheerful method (and our charges to external customers are about 1/2 to 1/3 of most other labs). Our main weakness is imaging small devices - an individual device below about 20um in size is very difficult to hit without high precision polishing techniques. If you're intending to do individual devices for failure analysis, then you will do better with the tripod polisher; otherwise, you may do better using the money to employ someone to make samples full-time. There are many stages in making a good specimen, and the best samples come with practice! We also make diamond TEM grids which are very useful for etching and ion-milling III-Vs and II-VIs which need good cooling.
Regards,
Richard Beanland GMMTL Caswell, Towcester, Northants NN12 8EQ UK Email richard.beanland-at-gecm.com Tel +44 1327 356363 Fax +44 1327 356775
Does anyone know of a procedure for tuning vertical wavelength dispersive spectrometers, specifically JEOL WD spectrometers? These are older, 2-crystal spectrometers on a 733.
Thanks
Mike Spilde Dept. of Earth & Planetary Science University of New Mexico Albuquerque, NM 87131 Electron Microprobe Laboratory 505/277-5430
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My thanks to all who replied to my question about leaving the filament control knobbed turned up to the saturation point on our JEOL 840A (with W filament). The consensus appears to be that there is no problem with the practice.
Note to Bill T., We have been leaving our filament current knob turned up over days on non-use and don't appear to be losing any filament life for it. Therefore, it appears that the current is turned off internally when the high voltage is turned off.
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G'day Would anybody in EM land know of the addresses and contact numbers of dealers in second hand (used) EM's and EM equipment anywhere, not just Australia. Reply directly to me if you can.
Huw Rosser CEMMSA The University of Adelaide South Australia
} Not true! } } The sputtering rate/cross-section at the SURFACE of a material } is not the same as the displacement rate/cross-section in the bulk. The } simple way to } think about this is that a surface you only have ~ 1/2 the number of bonds } that you have in the interior of a bulk. Thus the energy needed to remove a } surface atom is LESS than that needed to remove one from the bulk. There } is extensive literature on this unfortunatley, I don't have the references } readily at hand here at home. } Dear Nestor, In addition to removing the surface atom, do you not also create a new surface atom from a previously buried atom? If a single atomic layer were removed, wouldn't this be the equivalent of removing a single layer from the bulk and rejoining the atoms? I guess if you could remove an atom from the bulk, but not rearrange the remaining atoms, that would require more energy. Is that what you mean by "remove one from the bulk"? Yours, Bill Tivol
} I agree that magnetic media has a lifetime of 10 years or so, whereas } optical media is far longer, say 30 years.
Dear Larry, list, et. al,
I remember an article in Scientific American about a year ago which mentioned responsible estinates that were MUCH MORE PESSIMISTIC:
and on the order of 2 years for tape, 5 years for magnetic disk and about 10 years for CD.
Todays recording densities make for far more fragile records. in the "old days" ten years ago, the energy used to store each bit was 10's of 1000's (!!) times greater than that used today. RAM designers, as a paralell example, are discussing quantum limits ( 1 electron) of data cells.
This is NOT a simplistic topic, but one lying under layers and layers of systems design.
Regards,
Ed Monberg e-mail: {em-at-mediacity.com}
------------------------------------------------------ 510-429-1060 Fax 429-1065 LMDC, (Laser Motion Development Co.) 3101 Whipple Road Union City, CA 94587-1216
For our Most recent Catalogue of "On Hand" EQUIPMENT: Send empty mail to: {Cat-at-lasermotion.com}
} Now a question, } We have adopted the practice of leaving the filament current control on our } JEOL 840A at the proper saturation point (for our normal 15 kv operating } point) and just turning off the high voltage control for exchanging samples. } [snip] My question to } the list is whether there might be any adverse consequences from this } practice as there do not appear to be any.
Dear Warren, I assume the filament can be switched off with the current control in the appropriate position, as is the case for our EM's. If so, there will be no adverse effect for W, and presumably LaB6. If the filament is still drawing current with the HV off, there should only be the effect on filament lifetime equal to the number of cumulative hours of specimen exchange and the occasional disaster when the filament is not turned off over a weekend. Yours, Bill Tivol
"Home pages are the pet rock of the 90s. We all have them, we all think they're very cute. But in a few years we're going to look back and be pretty embarrassed." -- Tony Shepps {toad-at-pond.com}
Has anyone successfully "unstuck" Polaroid Type 55 negatives that have stuck together while drying? A water soak did not seem to help nor did Photoflo.
Thanks,
Dennis Shubitowski University of Michigan School of Dentistry
I just stumbled upon a graphical connection between electrostatic field components from Coulomb's law and the "line of no contrast" one finds in dark-field images of small strained crystal defects in TEM darkfield imaging. The connection is obvious "on reflection", but the experience of seeing what looks like a TEM image flowing from simple electrostatics calculations might be worth a visit, at least for folks who apply "physics filters" to TEM images, to my E&M course's triplet-visualization link at {http://newton.umsl.edu/~philf/triplet.html} .
Enjoy. /philf :)
//\/\/\/\---} // Phil Fraundorf Physics&Astronomy/CME 314-5165044 philf-at-newton.umsl.edu \\ B503 U.Missouri-SL St.Louis MO 63121 USA http://newton.umsl.edu/~philf \\/\/\/\/\/\/\/---}
There is a straightforward way of calibrating local specimen temperatue when using a heating holder:
D. C. Paine, D. J. Howard, and N. D. Evans, "In Situ TEM Studies of the Effect of Misfit Strain on the Kinetics of Si1-xGex Solid Phase Epitaxy: Temperature Calibration & Surface Effects," Electron Microscopy 1992: Proc. 50th Annual Meeting of the Electron Microscopy Society of America, G. W. Bailey, J. Bentley, and J. A. Small, eds., San Francisco Press, (1992), 1344-45.
To summarize, the thermocouple provides an accurate measure of the furnace temperature. However, actual specimen temperature, as a function of furnace temperature, will vary (at best) slightly from specimen to specimen. This is due to differences from specimen to specimen in the conduction path, that is, differences in contact resistance at the furnace/specimen interface as well as specimen geometry affects.
Specimens having cracks or other features which would restrict or alter heat flow will have greater uncertainty associated with actual temperatures.
I have been injecting cultured hepatocytes with antibodies and I am seeing interesting morphological changes on the LM. I now need to prepare injected cells for the TEM (not a problem). I am wondering what electron dense material(s) could be included in the injectate to show that a particular cell was injected (I want non-injected cells included on the coverslip as controls). The material must not interfere with good specimen appearance, and it also must be injectable (not toxic or clog my needles). Any help will be greatly appreciated!!!
Thanks in advance! Eugene Krueger Mayo Foundation GI Research krueger.eugene-at-mayo.edu
a user brought me a sample of gelatinous unicellular algae she preserved in 2% phosphate buffered glutaraldehyde. Unfortunately, it was a marine sample, so when she fixed it, the salts in the seawater precipitated. She can't repeat the experiment. Any way to salvage her sample--i.e., separate the cells from the ppt, or get the salts back into soln without detroying the cells? We appreciate that the precipitation may have ruined her cell preservation as well, but we won't know for sure till we can embed and section.
thanks in advance
steve
Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University San Diego, CA 92182-4614 phone: (619)594-4523 fax:(619)594-5676 email:sbarlow-at-sunstroke.sdsu.edu
Hi, Sorry for sending unsbscribe message for this list. But I couldn't unsubscribe by sending unsbscribe message to listserv-at-sparc5.microscopy.com. Could anybody tell me how can I unsubscribe? I also tried to get help from listserver by sending help command. But no response.
I'm really sorry for sending unsbscribe message to entire list.
Hiro. =========================================================== Hiroyuki Hakozaki Laboratory for Neurocytology Department of Neurosciences School of Medicine University of Calofornia , San Diego La Jolla , CA 92093-0608 Tel (619)-534-4583 or 2583 Fax (619)-534-7497 E-mail hiroyuki-at-ncmir.ucsd.edu ===========================================================
} Hi Mark; } } [snip] but are you sure you don't have } aluminum present? I would do a couple of things to check this; } } 1) Collect a "blank" spectrum of the matrix and see if you get a peak at } 1.49 keV. If you do, you either have some aluminum in the matrix,
To add a bit to Bob's reply, Br has an L-peak overlapping the Al K-peak. Although it is likely that a peak at ~1.49 keV is Al, check the 10-20 keV range for other peaks at the Br K alpha & beta positions. I have seen sediments with Al and no Br in one grain and Br in a nearby grain. Yours, Bill Tivol
If you are using a Mac, there are several different resources as part of NIH's Image freeware. I believe the best access is at http://rsb.info.nih.gov/nih-image/ This is a very good program for image manipulation for Macsters (that's why I'm posting to the list). A PC version is in the works too.
I know that I've seen stereo pairs, of the red/green variety, discused in the recent past. If there is nothing directly listed at NIH's web page there is an Image list that should have archives. The listproc address is LISTPROC-at-SOILS.UMN.EDU, and the list is NIH-IMAGE. (Cap.s not necessary.)
Erik ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Cretaceous Liability: Tyrannosaurus Wrecks
-- [ From: Charles A. Garber, Ph. D. * EMC.Ver #2.10P ] --
On Feb. 28, Sally Shrom wrote:
--Subject: LM and EM tech schools } } Are there any 2 year tech schools in the Philadelphia area? Sally Shrom
I would suggest you contact the following:
West Chester University West Chester, PA 19380
Attn: Prof. Arthur Smith Dept. of Geology
e-mail: asmith2-at-wcupa.edu
They have an outstanding program, Prof. Smith has been doing this for some years and his students have no difficulty finding jobs, even in today's difficult job climate.
Hope this helps. BTW, no connection here with West Chester University!
Chuck
====================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI SPI SUPPLIES FAX: 1-(610)-436-5755 PO BOX 656 e-mail: GVKM07A-at-prodigy.com West Chester, PA 19381-0656 USA Customer Service: SpiSupp-at-aol.com
In message {v02120d01ad5a491be151-at-[141.211.157.94]} Dennis Shubitowski writes: } Has anyone successfully "unstuck" Polaroid Type 55 negatives } that have stuck together while drying? A water soak did not } seem to help nor did Photoflo. } } Thanks, } } Dennis Shubitowski } University of Michigan } School of Dentistry } }
Dennis, Yes, I have successfully unstuck pairs of Polaroid negs. Actually, the water soak will work, but you have to soak them for a long time, at least overnight, say 12-16 hours. Then you have to slowwwwwwwly pull them apart to avoid pulling the emulsion off. A "pucker" mark may remain where they were joined, but usually its translucent so that a positive print will seldom reveal the pucker mark, especially on "busy" image detail, and as long as original image density was not disturbed.
When this happens to my negs, its usually in a spot no larger than the size of a quarter (25 cent piece). If its most of the area of the negs, that is serious trouble.
When I dry Polaroid negs in racks, I skip slots so that there is lots of space between adjacent negs to prevent sticking from occuring as the negs "wiggle" as they air dry.
--
Gib Ahlstrand, MMS Newsletter Editor Electron Optical Facility, University of Minnesota, Dept. Plant Pathology 495 Borlaug Hall, St. Paul, MN 55108 (612)625-8249 612-625-9728 FAX, giba-at-puccini.crl.umn.edu
"MICROSCOPY & MICROANALYSIS 96" in Minneapolis, Minnesota, Aug.11-15, 1996
} } I agree that magnetic media has a lifetime of 10 years or so, whereas } } optical media is far longer, say 30 years. } } Dear Larry, list, et. al, } } I remember an article in Scientific American about a year ago } which mentioned responsible estinates that were MUCH MORE PESSIMISTIC: } } and on the order of 2 years for tape, 5 years for magnetic disk and } about 10 years for CD. }
Also, you should read the 'fine print' when it comes to statements about media lifetime estimates. DAT DDS tapes, for example, have a 10 year estimated lifetime. What that actually means is that if you use a tape once to record and put it on a shelf in a temp. and humidity controlled environment, you are likely to be able to read it back in 10 years. Divide that by some factor if stored under less than ideal conditions. I was interested in archival use, rather than backups, where I may need to read back the data many times over that decade. Ask about how many passes of tape over the heads, including rewinding and forwarding, the tape is rated for, and you get a figure that may translate into only a few years of use (depending).
Data format is another problem. I think the ISO CD-ROM standard is in wide enough use to rely upon, but for magneo-optical disks, my experience has been that changing SCSI controller, operating system or other software, even ( with our Pinnacle MO drives ) chaning from one version of DOS to another, is enough to make the data effectively unreadable. ( The documentation necessary to write a program to retrieve that data is available, but not from Pinnacle -- we had to talk to their suppliers to get any useful information. ) I was also told by Pinnacle that they had changed suppliers for their SCSI controller cards, and if we bought a newer model, they would not guarantee that we would be able to read old data on the new drive. I don't know that Pinnacle is any worse that any other supplier -- that's just who I had my unhappy experience with. However, none of the salespeople will bring up the subject, and few will even understand your concerns, if you try to ask them how likely you are to be able to read your data 10 years from now.
ISO 9660 CD format and 'tar' files on DDS tapes (in that order) are the only formats in which I have much long term (decade) faith.
---| Steven D. Majewski (804-982-0831) {sdm7g-at-Virginia.EDU} |--- ---| Computer Systems Engineer University of Virginia |--- ---| Department of Molecular Physiology and Biological Physics |--- ---| Box 449 Health Science Center Charlottesville,VA 22908 |--- [ "The grass is always greener, except at t=0" - Stan Kelly-Bootle ]
{Text_1} Dennis Shubitowski asked "Has anyone successfully "unstuck" Polaroid Type 55 negatives that have stuck together while drying?"
I have had limited success with an overnight Photoflo soak followed by ultrasonication. Films stuck emulsion to film back seem to separate easier than those stuck emulsion to emulsion.
------------------------------------------------------------ Richard J. Mount Phone: 416-813-6551 Auditory Science Laboratory Department of Otolaryngology FAX: 416-813-5036 The Hospital for Sick Children 555 University Ave. Toronto, ON, Canada M5G 1X8
My family would be gratefull for suggestions involving the SEM. We are thinking about putting iron and stainless steel strips into various pH solutions to measure material loss and corrosion. SEM photos of the etch pits should make good illustrations.
Do you metallurgical/material science types have any suggestions to help? pH choices, time of exposure, how to introduce oxygen to enhance corrosion, etc.
Thanks, Joe Geller Geller MicroAnalytical Lab 426e Boston St. Topsfield, Ma 01983-1216 508 887-7000 fax 887-6671
On the subject of used EM equipment, we have an old Phillips TEM that we were considering putting up for sale. We were unsure whether there would be any demand for these instruments. (It needs a new condenser lens). I'm not sure of the model number, but I can get specifics if anyone is interested.
Cheri Owen Detroit Neurotrauma Institute Wayne State University Detroit, MI 313-577-4648
Hi All, I have a sample that I need some help with. We have received some samples that are from private homes. The residents are reporting a film (soot) forming on the wall above heat registers, on windows, and settling on furniture. These homes have newly installed high-effiency furnaces, and the question is whether the deposits are from the furnace, or some other source. We have analyzed some of the soot, and found it to be carbon with EDS. Now comes the tough part, is it possible to determine the source of this carbon (whether it comes from the burning of natural gas, a cigarette, or a candle). Any help in sampling and analyzing techniques would be greatly appreciated. Thanks,
I have successfully done this on a PC and Mac using Adobe Photoshop and 2 appropriately parallaxed images. Both must be changed to RGB color, the left (or zero degree) image saturated to red, and the other image saturated to either green or blue. Copy one image to the clipboard and then paste it to the other. Set the composite controls to 50% opacity with preview on. Viewing the image through red/green glasses, you can then properly register the pair.
It's a fairly crude technique, but it does work.
-=W.L. Steffens=- Department of Veterinary Pathology College of Veterinary Medicine University of Georgia
A colleague asked if I would submit this to the listserver.
snip.. } I am looking for replacement bulbs for 20-year old Vickers reflected light } petrographic microscopes. The bulbs we have been using are encoded } Phillips-Holland 13702C 6 volt 15 watt. Any help in locating such bulbs } will be greatly appreciated. snip..
Thanks in advance.
Owen Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
A colleague asked if I would submit this to the listserver.
snip.. } I am looking for replacement bulbs for 20-year old Vickers reflected light } petrographic microscopes. The bulbs we have been using are encoded } Phillips-Holland 13702C 6 volt 15 watt. Any help in locating such bulbs } will be greatly appreciated. snip..
Thanks in advance.
Owen Owen P. Mills Michigan Technological University Metallurgical & Materials Engineering Rm 512 MME Building Houghton, MI 49931 906-487-2002 906-487-2934 FAX opmills-at-mtu.edu
University of Central Florida Dept. of Mech. and Aero. Eng. Materials Science and Engineering Program
A M.S. or Ph.D. level graduate student is needed to perform research in the area of thin films with applications in electronic materials in collaboration with Lucent Technology (formerly AT&T Microelectronics). Research will be performed on Campus at the University of Central Florida, Orlando, and at the research facilities of Lucent also in Orlando. Research may begin in the Summer 1996 semester. Please address inquiries/resumes to:
************************************************************************* Lucille A. Giannuzzi, Ph.D. Assistant Professor
Materials Science and Eng. Program Dept. of Mechanical and Aerospace Eng. phone (407) 823-5770 University of Central Florida fax (407) 823-0208 4000 Central Florida Blvd. email lag-at-pegasus.cc.ucf.edu Orlando, FL 32816-2450 USA *************************************************************************
Subject: Time:3:40 PM OFFICE MEMO Mo aperture grids Date:02/29/96
Hi, I am looking for TEM grids with single hole of 0.8, 1.0 and 1.5 mm in diameter and slot of 1x2 and 0.5x2 mm. I tried several vender and did not succeed. Anybody know a source of these grids? Thank you in advance for your help.
Yi Huang Argonne National Lab/MSD 212 Argonne, IL 60439 tel: 708-252-5181 fax: 708-252-4798 yi_huang-at-qmgate.anl.gov
Subject: Time:3:40 PM OFFICE MEMO Mo aperture grids Date:02/29/96
Hi, I am looking for Mo TEM grids with single hole of 0.8, 1.0 and 1.5 mm in diameter and slot of 1x2 and 0.5x2 mm. I tried several vender and did not succeed. Anybody know a source of these grids? Thank you in advance for your help.
Yi Huang Argonne National Lab/MSD 212 Argonne, IL 60439 tel: 708-252-5181 fax: 708-252-4798 yi_huang-at-qmgate.anl.gov
Message-Id: {199602291508.JAA20746-at-mailhub.iastate.edu} X-Sender: wes-at-pop.ameslab.gov X-Mailer: Windows Eudora Pro Version 2.1.2 Mime-Version: 1.0 Content-Type: text/plain; charset="us-ascii"
Partitioning becomes a balancing act. There are two rules to remember that govern partitioning.
Rule 1 - You are allowed a maximum of 64K clusters per partition and each cluster has to be a power of 2 bytes long, hence 2048, 4096, 8192, etc.
Rule 2 - About half a cluster (on the average) is wasted per file.
Now the balancing act - If you will be storing lots of small files then small partitions are good in order to not waste too much space. But you want large enough partitions to do you some good. Of course, most EM image data files are quite large so that the number of clusters per file will be larger and the fractional wasted space less. You will want a large partition just to get a reasonable number of files on it.
There is also the hassle of trying to keep track of all the partitions. System and application files can reside on one partition, smaller data files on another, large image files on a third. But how do you use the remaining partitions? Perhaps they can be used to assign a partition to each user. However, I still have a PDP-11 system with a 170 MB drive (it was big back then) which *had* to be partitioned into segments no larger than 32 MB. We distributed users around as space was availabe. But it was hard to remember where we put them without a good search utility (which we used).
Practically, we have three 850 MB drives in our group that I have partitioned into 4 pieces each. We defined 1 127 MB segment for smaller data files, and split the remainder up into 3 partitions of about 240 MB each; one is used for system files, one for large images, and the third is used for temporary projects but is mostly empty.
Hope this helps. BTW, Partition Magic caught my eye the other day. It would be nice to adjust such things on the fly.
At 11:06 AM 2/28/96 -0800, you wrote: } } I have been following the CD thread etc. I thought someone might be } interested in this: Bought a notebook with 800MB HD and the usual installed } software taking up about 60 MB. Using Partition Magic 2.0, I was able to } repartition the HD on-the-fly and created 500MB and 250MB partitions (42 MB } reserved for suspend mode). In doing so, I recovered an extra 10 MB due to } the smaller sector sizes used. That's a nice feature of PM in that it can } change sector sizes on- the-fly. What I mean by "on-the-fly" is that you } don't have to backup the HD, re-partition it, and then reinstall the } software; you just run the software, create whatever partitions and sizes } you want, and you are done. It can do a lot more than that but you can get } information from the vendor. } } Since the sector size changes with partition size (the break points are at } 128, 256, and 512 or some such numbers), has anyone partitioned a large HD } into 128 MB drives for the greatest storage efficiency or are there down } sides to that? Would be interested in hearing your thoughts on that. } } Disclaimer: I have no financial interest in PowerQuest (mfg. of Partition } Magic). } } Damian Neuberger } neuberd-at-baxter.com } ---------------------------------------------------- Warren E. Straszheim 270 Metals Development, Ames Lab/ISU, Ames IA, 50011 Phone: 515-294-8187 FAX: 515-294-3091
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I worked with an electrophysiologist who used procion brown. Didn't seem to harm the ultrastructure but did make the cells more electron dense than the surrounding retinal neurons. Ken-iche Naka would stick and record retinal neurons with a dye loaded micropipette. Once the recording was done, then he injected the dye into that cell. Recovering the cells was a bit arduous.
} Hello All } } Help! } } I have been injecting cultured hepatocytes with antibodies and I am seeing } interesting morphological changes on the LM. I now need to prepare injected } cells for the TEM (not a problem). I am wondering what electron dense } material(s) could be included in the injectate to show that a particular cell } was injected (I want non-injected cells included on the coverslip as } controls). The material must not interfere with good specimen appearance, } and it also must be injectable (not toxic or clog my needles). Any help will } be greatly appreciated!!! } } } } Thanks in advance! } Eugene Krueger } Mayo Foundation } GI Research } krueger.eugene-at-mayo.edu
Charles J. Butterick (Chuck) Electron Microscopy Center Department of Cell Biology and Biochemistry Texas Tech University Health Sciences Center 3601 4th Street Lubbock, Texas 79430
vox (806) 743-1633 fax (806) 743-1219 email emccjb-at-ttuhsc.edu or chuck-at-micron1.lubb.ttuhsc.edu
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