The H7000 that I use has another exposure meter display inside the column just behind the screen. this consists of two red LEDs (ie no green middle. If the two come on together that is a correct exposure but increasing or decreasing the beam density on the screen should make one or the other come on. I apologise if you have checked this but if the column meter works then it sounds like it is the other meter. Also have you checked that no one has 'mucked about' with the meter calibration settings.
If everything else checks out then maybe it's just the link from the central screen that has broken - remember this is a current density rather than light meter.
I hope this helps.
Malcolm
Malcolm Haswell e.m. unit Chemispec School of Health, Natural and Social Sciences Fleming Building* University of Sunderland Tyne & Wear SR1 3SD UK tel no: 0191 515 2872 / 3468 e-mail: malcolm.haswell-at-sunderland.ac.uk
----- Original Message ----- } From: Garry Burgess {GBurgess-at-exchange.hsc.mb.ca}
Dear Sir/Madam, I am a Private Investigator based in Europe. A group of Government Officials from an African Country contacted me with a Proposal. I am to Make contact with you and state their offer, if your Interest is Genuine, you will be contacted for your Account details to which will be transferred the sum of $33,600,000.00 USD. (20% of which is yours). You are then required to forward the remaining balance (Minus the Interest, handling and tax clearance charges, which Will be offset by Us & Deducted from the transferred sum) to a nominated Bank account in the Cayman Islands. I don't think I need to spell out the importance of Secrecy in this Matter considering the amount involved.
Let me state clearly here that the account that you would be providing does not need to have funds in it; it is only needed to be active and be able to receive funds. So, if I don't hear from you within three days I will assume you are not interested and will solicit for a new partner, but if you know you are interested let me know. List your phone & fax Numbers so we may communicate with you. This is important as we would Have to talk about the modalities of the transaction. Waiting to hear from you, Abiodun John
please do reply to my alternate and secure email abiodunjohn-at-lycos.co.uk
Hi Randy Do you have the same problem when you use gold or nickel grids? Some time ago I worked on immunogold technique in tissue and I had problem with pepper/dirt film over the sections. I used nickel grids that were bought ages ago. I bought a new batch, cleaned it and sections were clean, no pepper/dirt film. Dorota
Hi all, I have 2 standard cubes I use (blue: excites at 490, emits at 515, for chlorophyll a, and green: excites at 545 and emits at 590 for phycoerythrin/cryptomonads), plus I have an additional cube for UV that I use for DAPI and calcafluor white. My problem is taxa that are from humic stained lakes. Does anyone have a better Excite/Emission range suggestion for these systems, especially for phycoerythin or cryptomonads? Pigments are measuring high, cells look healthy and are well colored, but fluoresce poorly. I think the pigments have shifted their absorption ranges, rather than the humics interfering (The preps I'm working with are very thin, with no overlying water). This has been troublesome for a while, but lately, I seem to be getting lots of samples from humic systems and I can ID and count much faster if I have good, active fluorescence. I'll post responses to the list if folks are interested. Thanks! Ann.
Ann St. Amand, Ph.D. President PhycoTech, Inc. 620 Broad St., Suite 100 St. Joseph, Michigan 49085 USA
I am having some trouble embedding arabidopsis seedling for TEM. I am using epon-araldite mixture and my tissue just dissolves away upon sectioning of my samples. I am assuming it is an embedding problem. I dehydrate my samples using an ETOH gradient then switch to 100% propylene oxide. I embed with a 2:1 (PO:epon-araldite) mixture for 12 hours then a 1:2 mixture for 12 hours then 100% epon-araldite at room temp followed by 24 hours at 30 degrees then 48 hours at 45 degrees. Should I change something in my protocol or is there another embedding medium that may work better for arabidopsis seedlings? Any help would be appreciated. Thankyou.
Becky- I gave a microscope talk to my daughter's 4th grade class. On a Friday, I gave the teacher 24 plastic petri dishes, one for each student and the teacher as well. Then I had my daughter collect them on Monday. The rule was that - 1) no liquids, and 2) each dish had to be scotch-taped closed, with only the student's name on the label. The kids were free to collect a small specimen of anything animal (not living), vegetable, or mineral. I then made slides of each item, and tried to guess what each was. Some were obvious. I came back with my microscope, and the slides, and made handouts for each child (and the teacher). The 24 page handout had a microphotograph of each slide and the student's name under the relative picture. The hand out also had a diagram of the microscope, and some brief information (4th grade level). I let the students come up one, by one to look through the microscope at this or that more interesting specimen. To save money on the booklet, I made two sets of color prints: One booklet contained all color photos (for the teacher's copy) and I Black and White photocopied the book (in photo-mode) for the other 22 student copies. I used gluestick to glue an original color print of each student's specimen in their respective booklet, and handed them out accordingly. It was great fun, and it worked out perfectly, and didn't take up too much of the teacher's time. Total was about an hour total class time. The students were very creative in their choices, and brought a full spectrum of things microscopic into the class! Rgds, Mike Shaw
Question: I will be teaching gifted 5th graders in a new math/science program this year and need to know what type of microscope would be best for my program. Also, where can I buy a microscope for a reasonable price? Do colleges ever donate used microscopes to schools? I am working with a shoestring budget of $0.00 so I will have to make a personal purchase in order for my students to use this wonderful teaching tool. Thank you for asssisting me with this matter. I want to offer the best science program possible! Becky Hogan } }
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sconnell-at-liai.org) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, August 1, 2003 at 15:58:16 ---------------------------------------------------------------------------
Email: sconnell-at-liai.org Name: samuel connell
Organization: La Jolla Institute for Allergy and Immunology
Education: Graduate College
Location: San Diego, CA
Question: Iím heading into confocal microscopy calcium flux land for the first time. Iíve done a fair amount in flow cytometry using Indo-1. Iím interested in speaking with someone who has experience in doing this work with visible wavelength probes, preferably with the BioRad MRC 1024 system. Our system has the Kr/Ar mixed gas with the 488, 568, and 647 laser lines available. Weíre planning on looking at 512x512 XY images over time, potentially with scans every 5-10 seconds over 10-20 minutes or so. If someone would be so kind as to touch base with me, I would be truly indebted for the shared expertise. Thanks, -- Samuel Connell
We've done some work with Ca green. Basically, it gets brighter and dimmer depending on the free Ca. It works with blue excitation. Not particulary quantitative, but it gets the point across.
I know that Gatan makes various vacuum transfer holders for the TEM but are there any other suppliers?
Many thanks,
Ralph Haswell Research Scientist Shell International Chemicals B.V. Shell Research & Technology Centre, P.O. Box 38000, 1030 BN Amsterdam, The Netherlands
I really am wondering why nobody has raised this important question earlier. Yes, RGB color interpolation with Bayer filter cameras can become an problematic factor when it comes to quantitative spatial measurements of coloured samples. Especially when working with low power magnification where camera resolution is crucial.
Regarding Bayer filter cameras, you might want to consider the pixel shifting technology used in the ProgRes C14 digital microscopy camera from JENOPTIK Laser, Optik, Systeme GmbH.
The ProgRes C14 also uses a single CCD with Bayer filter design. The sensor can be moved by means of a piezoelectric shifting device as many other pixel shifting cameras use.
But there are different ways to shift the pixels to produce new information. Most pixel shifting camera use the sensor shifting only to gain optical resolution, i.e. scanning the image more accurately, which makes much sense when using high resolution optics. Thereby the sensor is shifted in fractions of the pixels' spacing, producing a slight overlap of the pixels. But: quantitative spatial measurements might become problematic due to the RGB color interpolation, where some spatial resolution is defintively lost.
In contrast to some other pixel shifting cameras, JENOPTIK's ProgRes C14 also shifts the sensor in full pixel steps, where the pixels overlap 100%. Doing this at least four times in x- and y- direction you sample the full RGB color information for each single pixel position. In the result you get a true color RGB image without interpolation, comparable to a RGB filter wheel camera. The file size (and thus nominal image resolution) stays the same. But precise quantitative spatial measurements of colored samples is now possible. We call this technique "Color-Co-Site-Sampling" and as far as I know only ZEISS's Axicam HRc and JENOPTIK's ProgRes C14 do use this technique in micro-photography.
You can combine this full pixel shifting increasing "color resolution" also with fractional pixel shifting increasing spatial resolution.
An image says more than a thousand words - so if you want some illustration for this technique please contact me.
The benefit of this technique compared to a filter wheel camera is, that you can use the camera also for moving specimens in single shot mode, where colors then are interpolated. And compared to a 3 CCD camera, the Bayer filer camera is more light sensitive, because incoming light does not need to be split up by a prism directing it onto the three single CCDs.
Of course, also in pixel shifting cameras pixel misalignment can occur - in principle. You need to keep in mind that the sensor shifting has do be done very precisely in the sub-µm range. To guarantee optimum alignment, the ProgRes C14 comes with a built-in calibration procedure that allows the user to calibrate the piezo scanner for best results.
I hope this information does add a new facette to your discussion about quantitative spatial measurements of coloured samples with Bayer filter cameras.
} -----Original Message----- } From: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] } Sent: Thursday, July 24, 2003 10:52 AM } To: MSA } Subject: Bayer filter versus 3CCD camera } ---------. } } } Hi, } } The two main types of color cameras used in microscopy I know } of are the } ones with 3 separate CCDs for each of the three primary colors and the } ones with a Bayer filter design. In the 3CCD camera the } spatial sampling } is the same for each of the three colors, but not in the Bayer filter } camera design ? How do you deal with this difference when doing } quantitative spatial measurements of colored samples in microscopy ? } } What is the relative actual resolution seen with a 3CCD color } camera and } a camera with Bayer filter reconstruction? How to take into } account the } reduced sensitivity of the Bayer filter type for each color, compared } with the 3CCD camera if any ? When are Bayer filter type } cameras a valid } alternative for a 3CCD camera in microscopy, besides the price ? } } What about pixel shifts in a 3CCD camera due to misalignment of the } color filters/splitter and the CCDs ? } } Best regards, } } Peter Van Osta } } Union Biometrica N.V./S.A. } European Scientific Operations (ESO) } Cipalstraat 3 } B-2440 Geel } Belgium } Tel.: +32 (0)14 570 619 } Fax.: +32 (0)14 570 621 } } http://www.unionbio.com/ } } http://ourworld.compuserve.com/homepages/pvosta/cvwww.htm }
Hi, My department is currently considering to buy a low voltage ion mill. The users I've spoken to up to date all agree concerning the good quality of the samples. However, the maintenance of many low energy ion
mills seems be a major problem, especially in the case of a large number
of users. We are also concerned about technical support from suppliers located overseas. Does anybody have experience with suppliers like Fischione and South Bay
Technology with respect to maintenance and technical support in Europe? Certainly, I am also interested in comments on the effectiveness of low energy ion mills in general.
Bj Health Care is a major problem in the U.S.... Insurance companies have raised premiums 30% or more and businesses are cutting back on giving healt insurance...
Walking around without any health care protection is like playing Russian roulette with yourself and your family.
Click Here For A FREE Health Care Consultation. {http://www.nationalhealthcareplan.com/click.html?aid=633&cid=1341}
National Health Care Plan's Complete Care Plan is designed for the following:
» Everyone Gets Accepted! » Pre-Existing Conditions are OK!!! » Personal or family emergency's ($3000 Accident Protection) » Discounts up to 80% on medical services » Discounts up to 80% off on prescription drugs » Dental exams & X-Rays at no charge » Doctor Visits & Chiropractic Care as Low as $20 » Eye Exams only $35
Stop Receiving the offers {http://www.medsforlife.biz/removes.html}
FIRST AND FOREMOST,I MUST SOLICIT YOUR STRICTEST CONFIDENCE IN THIS TRANSACTION AND I PRAY THAT MY DECISION TO CONTACT YOU WILL BE GIVEN GENUINE APPROVAL CONSIDERING THE FACTS WE HAVE NOT KNOWN EACH OTHER BEFORE, I WISH TO USE THIS OPPORTUNITY TO INTRODUCE MYSELF TO YOU.
I AM PRINCE JOHNSON FROM LIBERIA,WEST AFRICA. I WRITES TO INFORM YOU MY DESIRE TO INVEST,AND TO BUY A LIVING HOUSE IN YOUR COUNTRY. I AM THE FIRST SON OF MR. WEAH JOHNSON, HE WAS A DIAMOND/GOLD MERCHANT IN MY COUNTRY.MY FATHER HAD A BULLET SHOT BY THE REBELS ON HIS WAY TRAVELLING OUT OF MY COUNTRY WITH TWO OF MY YOUNGER SISTER'S DUE TO PRESENT CRISIS THAT IS OCCURING IN MY COUNTRY(LIBERIA).MY SISTER'S DIED ON THE SPOT WHILE U.N.PEACE KEEPING FORCE RESCUED MY FATHER,HE WAS TAKEN TO HOSPITAL FOR MEDICAL TREATMENT WHICH HE LATER DIED. BEFORE HE DIED HE REAVEALED TO ME AND MY MOTHER ABOUT THE BOXES CONTAINING $8 MILLION US DOLLARS.WHICH HE DEPOSITED WITH A SECURITY COMPANY IN GHANA FOR SAFE KEEPING. MY FATHER DID NOT DISCLOSE THE CONTENT OF THE BOXES TO THE SECURITY COMPANY.TO AVIOD THE OFFICIALS FROM RAISING EYE BROWS TO THE FUNDS.
PRESENTLY MYSELF AND MY MOTHER ARE HERE IN GHANA TO NOTIFY THE SECURITY COMPANY FOR THE CLAIMS,AND WE ARE STAYING IN THE REFUGEE CAMP. THEREFORE I WANT YOU TO LECTURE ME ON HOW BEST WE CAN INVEST THIS MONEY,BECAUSE MY FATHER TOLD ME THAT IT IS DANGEROUS TO INVEST THIS MONEY IN AFRICA TO AVIOD SUSPICIONS,AND DUE TO MARKET INSTABILITY COUPLED WITH ECONOMIC AND POLITICAL INSTABILITY FACING AFRICAN COUNTRIES,THAT IS WHY WE WANT TO INVEST IN ABROAD. FOR YOUR MUTUAL ASSISTANCE, MYSELF AND MY MOTHER HAVE AGREED TO OFFER YOU 20%OF THE TOTAL AMOUNT OF THE MONEY.
WE HAVE ALL THE VITAL DOCUMENTS COVERING THE DEPOSIT AND THE OWNERSHIP WHICH I CAN SEND TO YOU THROUGH FAX ON REQUEST. NOTE:I HAVE NEVER DISCLOSED THIS TO ANY PERSON APART FROM YOU,SO YOU HAVE TO KEEP THIS TRANSACTION AS A TOP SECRET TO YOURSELF ALONE.WHICH I WILL WANT YOU TO FORWARD ACROSS TO ME YOUR DIRECT TEL/FAX NUMBER FOR MORE INFORMATIONS ABOUT THIS TRANSACTION.
BEST REGARDS,
PRINCE JOHNSON. (FOR THE ENTIRE FAMILY NOTE:YOU CAN ALSO REACH ME ON MY ALTERNATIVE EMAIL BOX:prince_2003-at-myself.com
FIRST AND FOREMOST,I MUST SOLICIT YOUR STRICTEST CONFIDENCE IN THIS TRANSACTION AND I PRAY THAT MY DECISION TO CONTACT YOU WILL BE GIVEN GENUINE APPROVAL CONSIDERING THE FACTS WE HAVE NOT KNOWN EACH OTHER BEFORE, I WISH TO USE THIS OPPORTUNITY TO INTRODUCE MYSELF TO YOU.
I AM PRINCE JOHNSON FROM LIBERIA,WEST AFRICA. I WRITES TO INFORM YOU MY DESIRE TO INVEST,AND TO BUY A LIVING HOUSE IN YOUR COUNTRY. I AM THE FIRST SON OF MR. WEAH JOHNSON, HE WAS A DIAMOND/GOLD MERCHANT IN MY COUNTRY.MY FATHER HAD A BULLET SHOT BY THE REBELS ON HIS WAY TRAVELLING OUT OF MY COUNTRY WITH TWO OF MY YOUNGER SISTER'S DUE TO PRESENT CRISIS THAT IS OCCURING IN MY COUNTRY(LIBERIA).MY SISTER'S DIED ON THE SPOT WHILE U.N.PEACE KEEPING FORCE RESCUED MY FATHER,HE WAS TAKEN TO HOSPITAL FOR MEDICAL TREATMENT WHICH HE LATER DIED. BEFORE HE DIED HE REAVEALED TO ME AND MY MOTHER ABOUT THE BOXES CONTAINING $8 MILLION US DOLLARS.WHICH HE DEPOSITED WITH A SECURITY COMPANY IN GHANA FOR SAFE KEEPING. MY FATHER DID NOT DISCLOSE THE CONTENT OF THE BOXES TO THE SECURITY COMPANY.TO AVIOD THE OFFICIALS FROM RAISING EYE BROWS TO THE FUNDS.
PRESENTLY MYSELF AND MY MOTHER ARE HERE IN GHANA TO NOTIFY THE SECURITY COMPANY FOR THE CLAIMS,AND WE ARE STAYING IN THE REFUGEE CAMP. THEREFORE I WANT YOU TO LECTURE ME ON HOW BEST WE CAN INVEST THIS MONEY,BECAUSE MY FATHER TOLD ME THAT IT IS DANGEROUS TO INVEST THIS MONEY IN AFRICA TO AVIOD SUSPICIONS,AND DUE TO MARKET INSTABILITY COUPLED WITH ECONOMIC AND POLITICAL INSTABILITY FACING AFRICAN COUNTRIES,THAT IS WHY WE WANT TO INVEST IN ABROAD. FOR YOUR MUTUAL ASSISTANCE, MYSELF AND MY MOTHER HAVE AGREED TO OFFER YOU 20%OF THE TOTAL AMOUNT OF THE MONEY.
WE HAVE ALL THE VITAL DOCUMENTS COVERING THE DEPOSIT AND THE OWNERSHIP WHICH I CAN SEND TO YOU THROUGH FAX ON REQUEST. NOTE:I HAVE NEVER DISCLOSED THIS TO ANY PERSON APART FROM YOU,SO YOU HAVE TO KEEP THIS TRANSACTION AS A TOP SECRET TO YOURSELF ALONE.WHICH I WILL WANT YOU TO FORWARD ACROSS TO ME YOUR DIRECT TEL/FAX NUMBER FOR MORE INFORMATIONS ABOUT THIS TRANSACTION.
BEST REGARDS,
PRINCE JOHNSON. (FOR THE ENTIRE FAMILY NOTE:YOU CAN ALSO REACH ME ON MY ALTERNATIVE EMAIL BOX:prince_2003-at-myself.com
THE DIRECTOR, AUDIT AND ACCOUNTS UNIT, FOREIGN REMITTANCE DEPT., INTERNATIONAL BANK FOR AFRICA, LOME,REPUBLIC OF TOGO,WEST AFRICA.
ATTN:
WITH DUE HONOUR AND RESPECT,I AM MR JACKSON BENSON,THE DIRECTOR
IN CHARGE OFAUDIT AND ACCOUNTS UNIT,FOREIGN REMITTANCE DEPT.OF THE INTERNATIONAL BANK OF AFRICA LOME-TOGO IN WEST AFRICA.
I GOT YOUR E-MAIL ADDRESS RECOMMENDED BY A TOGOLAISE BUSINESS CONSULTANT,ELDER JOHN KAFUI AND I DECIDED TO CONTACT YOU FOR BENEFICIAL AND A 100% RISK-FREE BUSINESS TRANSACTION. DURING OUR AUDITING AND INVESTIGATIONS IN THIS BANK,MY DEPARTMENT CAME ACROSS THE SUM OF THIRTY MILLION UNITED STATES DOLLARS (US$30,000,000)ONLY BELONGING TO A JAPANESE INTERNATIONAL BUSINESSMAN WHO DIED ALONG WITH HIS NEXT OF KIN IN THE 5TH NOVEMBER,1997 AEROPLANE CRASH IN ABIDJAN. BEFORE OUR DISCOVERY OF THIS DEVELOPMENT,THERE WAS NO TRACE OF CLAIM FROM ANY PERSON AS THE FUND REMAINS DORMANT IN HIS ACCOUNT WITH THIS BANK.
ALTHOUGH,I KEEP THIS INFORMATION SECRET WITHIN MY JURISDICTION TO ENABLE US PUT CLAIMS AND TRANSFER THE SAID AMOUNT THROUGH A TRUSTWORTHY FRIEND OVERSEAS WHOM WE SHALL PRESENT TO THE BANK AS THE BONAFIDE NEXT-OF-KIN TO THE DECEASED FOR A PROFITABLE AND SUCCESSFUL DEAL. MEANWHILE,ALL THE ARRANGEMENTS TO PUT CLAIMS AS THE BONAFIDE NEXT-OF-KIN TO THE DECEASED,TO GET THE REQUIRED APPROVAL AND TRANSFER OF THIS MONEY TO A FOREIGN ACCOUNT HAS BEEN PUT IN PLACE.THE DIRECTIVES AND THE NEEDED INFORMATION WILL BE RELAYED TO YOU AS SOON AS YOU INDICATE YOUR INTEREST AND WILLINGNESS TO BENEFIT YOURSELF FROM THIS GREAT BUSINESS OPPORTUNITY. INFACT,WE COULD HAVE DONE THIS DEAL ALONE
BUT BECAUSE,AS CIVIL SERVANTS WE ARE NOT LEGALLY ALLOWED TO OPERATE FOREIGN ACCOUNT.AND IT WOULD EVENTUALLY RAISE EYEBROWS ON OUR SIDE DURING THE TIME OF TRANSFER BECAUSE WE ARE STAFF OF
THE BANK.THESE ARE THE ACTUAL REASONS WHY IT REQUIRES A SECOND-FELLOW WHO WILL FORWARD CLAIMS BY OUR SUPPORT AS THE BONAFIDE NEXT-OF-KIN WITH TOGOLAISE'S COURT AFFIDAVIT TO THE BANK AND ALSO PRESENT A FOREIGN BANK ACCOUNT WHERE THE MONEY ON
HIS/HER REQUEST WILL BE RE-TRANSFERED INTO.
ON CONCLUSION OF THIS TRANSACTION,YOU WILL BE ENTITLED TO 25% OF THE TOTAL SUM AS GRATIFICATION.5% OF THE TOTAL SUM WILL BE USED TO REINBURSE EXPENSES THAT MIGHT ARISE FROM TELEPHONE BILLS AND OTHER EXPENSES DURING THE TRANSACTION,WHILE 70% WILL BE FOR ME AND MYPARTNERS HERE. PLEASE YOU HAVE BEEN ADVICED TO KEEP TOP SECRET AS WE ARE STILL IN SERVICE AND INTEND TO RETIRE
FROM SERVICE AFTER WE CONCLUDE THIS DEAL WITH YOU. I WILL BE MONITORING THE WHOLE SITUATION HERE IN THIS BANK UNTIL YOU CONFIRM THE MONEY IN YOUR ACCOUNT.WE THEN COME DOWN TO YOUR COUNTRY FOR SUBSEQUENT SHARING OF THE FUND ACCORDING TO THE PERCENTAGES PREVIOUSLY INDICATED AND FOR INVESTMENT IN ANY COUNTRY YOU MAY ADVICE US TOO. ALL OTHER NECESSARY INFORMATION WILL BE SENT TO YOU WHEN I HEAR FROM YOU. I SUGGEST YOU GET BACK TO ME AS SOON AS POSSIBLE,STATING YOUR WISH IN THIS DEAL.
YOURS FAITHFULLY, MR JACKSON BENSON. 00228-9118757
________________________________________________ Get your own "800" number Voicemail, fax, email, and a lot more http://www.ureach.com/reg/tag
Nestor Is there anything that can be done about the volume of SPAM this board sends to my inbox? I don't really want to, as this is a useful resource, but I am considering unsubscribing because of it. Cheers Ady
__________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com
I have started using a free spam filter called "Spam Assasin" or SAproxy. It was rated best by Consumer Reports. It is available from Blomba.com. For a free thing it is doing a fairly good job. I would guess that my spam has been cut by 70%. (now I feel alone and unwanted)
Greg
At 11:36 AM 8/6/2003 -0700, ady jenkinson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Gregory W. Erdos Ph.D. Assistant Director, Biotechnology Program P.O. Box 118525 217 Carr Hall University of Florida Gainesville, FL 32611 gwe-at-ufl.edu 352-392-1295
With all due respect and thanks to the manager of this extremely valuable list server, would it be possible to put a filter on the e-mail sent out? It should be possible for the list server to except mail ONLY from registered users. This list has become my number one source of spam (I am not interested in all of these people who wish to give me lots of money).
Thank you
David ____________________
David Elliott Ph.D.
Yale University School of Medicine Campus Address: TAC S160 333 Cedar Street PO Box 208022 New Haven, CT 06520-8022
Hmmm. I get maybe one spam a day from the list. Nothing compared to what is out there.
geoff
ady jenkinson wrote:
} Nestor } Is there anything that can be done about the volume of } SPAM this board sends to my inbox? } I don't really want to, as this is a useful resource, } but I am considering unsubscribing because of it. } Cheers } Ady } } __________________________________ } Do you Yahoo!? } Yahoo! SiteBuilder - Free, easy-to-use web site design software } http://sitebuilder.yahoo.com } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
The Listserver has a spam filter on it, and while admittedly a few messages get through it is a very small fraction of the "attempts".
Can you please check and insure that the spam is really coming from the Listserver and is not being "faked". This is becoming a common tactic among spammers. They find a real address and subsitute it for the address in the FROM field to make it appear that mail is coming from a legit address.
I saw 2 simultaneous spams that got through the filter yesterday (and they are now blocked) but nothing else for the week.
Just for the record in the last 48 hours there were 162 spam messages blocked by the filter. This does not count the 2 that got through.
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hmmm. I get maybe one spam a day from the list. Nothing compared to what } is out there. } } geoff } } ady jenkinson wrote: } } } Nestor } } Is there anything that can be done about the volume of } } SPAM this board sends to my inbox? } } I don't really want to, as this is a useful resource, } } but I am considering unsubscribing because of it. } } Cheers } } Ady } } } } __________________________________ } } Do you Yahoo!? } } Yahoo! SiteBuilder - Free, easy-to-use web site design software } } http://sitebuilder.yahoo.com } } } } } } } }
I wonder if anyone knows where we could find highly crystalline fine graphite powder with a platelet size in the range of 50-100nm (smaller is still OK). I have tried to grind an old AFM HOPG crystal and lost all my optimism after 2 hours...
You've been doing a great job on keeping the filter "tuned", keep up the good work.
Fellow Listers:
To complain about one or two spam messages here and there seems unrealistic. As a system administrator, I can say that the volume of spam, and the various tactics (and sophistication of them) that are used by the spammers, is constantly increasing.
The miniscule volume of spam on this list, and high signal to noise ratio, is a testament to Nestor's vigilance against this plague.
John W. Raffensperger, Jr. IS Manager Helwig Carbon Products, Inc.
Colleagues
The Listserver has a spam filter on it, and while admittedly a few messages get through it is a very small fraction of the "attempts".
Can you please check and insure that the spam is really coming from the Listserver and is not being "faked". This is becoming a common tactic among spammers. They find a real address and subsitute it for the address in the FROM field to make it appear that mail is coming from a legit address.
I saw 2 simultaneous spams that got through the filter yesterday (and they are now blocked) but nothing else for the week.
Just for the record in the last 48 hours there were 162 spam messages blocked by the filter. This does not count the 2 that got through.
Dear Sir, I am MALIK ALLI from Iraq.I decided to write you due my situation now.There is an information I would like you to keep very confidential.
There is sum amount of money my boss deposited in Europe, out side our country,and followiong the battle at Basra by the American Millitary forces,he was killed .It is only me that knows these because I'm his second,I will not be {BR} able to give you the full details that led to that urgly incidents during the War. Presently I am staying in another country,just to save my life,with my three children. The money in question, is Twentyfive million five hundred thousand united state dollars (40.500,000.00Milllion) U.S.Dollars. What I would want you to do, is to assits me to recouver the money as it can only be identified only by calling the pin no to the company concerned, There is no risk in this transaction. I will use the money for an investiment in your country for the future of my children. If you are intrested, and can maintain the very confidentiality of this transaction,you e-mail me on email;bashiralli-at-123.com immediately for more clearification,and also note that I am a refugees now since the americans've taken over our country and as a soldier,I'll not come back now becuase of the the situation in Iraq.I can speak little English,so dont mind my English. Thank you very much.
MALIK ALLI
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
Dear Sir, I am MALIK ALLI from Iraq.I decided to write you due my situation now.There is an information I would like you to keep very confidential.
There is sum amount of money my boss deposited in Europe, out side our country,and followiong the battle at Basra by the American Millitary forces,he was killed .It is only me that knows these because I'm his second,I will not be {BR} able to give you the full details that led to that urgly incidents during the War. Presently I am staying in another country,just to save my life,with my three children. The money in question, is Twentyfive million five hundred thousand united state dollars (40.500,000.00Milllion) U.S.Dollars. What I would want you to do, is to assits me to recouver the money as it can only be identified only by calling the pin no to the company concerned, There is no risk in this transaction. I will use the money for an investiment in your country for the future of my children. If you are intrested, and can maintain the very confidentiality of this transaction,you e-mail me on email;bashiralli-at-123.com immediately for more clearification,and also note that I am a refugees now since the americans've taken over our country and as a soldier,I'll not come back now becuase of the the situation in Iraq.I can speak little English,so dont mind my English. Thank you very much.
MALIK ALLI
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
Dear Sir, I am MALIK ALLI from Iraq.I decided to write you due my situation now.There is an information I would like you to keep very confidential.
There is sum amount of money my boss deposited in Europe, out side our country,and followiong the battle at Basra by the American Millitary forces,he was killed .It is only me that knows these because I'm his second,I will not be {BR} able to give you the full details that led to that urgly incidents during the War. Presently I am staying in another country,just to save my life,with my three children. The money in question, is Twentyfive million five hundred thousand united state dollars (40.500,000.00Milllion) U.S.Dollars. What I would want you to do, is to assits me to recouver the money as it can only be identified only by calling the pin no to the company concerned, There is no risk in this transaction. I will use the money for an investiment in your country for the future of my children. If you are intrested, and can maintain the very confidentiality of this transaction,you e-mail me on email;bashiralli-at-123.com immediately for more clearification,and also note that I am a refugees now since the americans've taken over our country and as a soldier,I'll not come back now becuase of the the situation in Iraq.I can speak little English,so dont mind my English. Thank you very much.
MALIK ALLI
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
Dear Sir, I am MALIK ALLI from Iraq.I decided to write you due my situation now.There is an information I would like you to keep very confidential.
There is sum amount of money my boss deposited in Europe, out side our country,and followiong the battle at Basra by the American Millitary forces,he was killed .It is only me that knows these because I'm his second,I will not be {BR} able to give you the full details that led to that urgly incidents during the War. Presently I am staying in another country,just to save my life,with my three children. The money in question, is Twentyfive million five hundred thousand united state dollars (40.500,000.00Milllion) U.S.Dollars. What I would want you to do, is to assits me to recouver the money as it can only be identified only by calling the pin no to the company concerned, There is no risk in this transaction. I will use the money for an investiment in your country for the future of my children. If you are intrested, and can maintain the very confidentiality of this transaction,you e-mail me on email;bashiralli-at-123.com immediately for more clearification,and also note that I am a refugees now since the americans've taken over our country and as a soldier,I'll not come back now becuase of the the situation in Iraq.I can speak little English,so dont mind my English. Thank you very much.
MALIK ALLI
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
I was a little surprised by the recent requests for spam filtering, and the threats to unsubscribe. Then I realized that they must be newer members. Those of us that have been around for a while, know the excellent job you do at keeping unwanted traffic to a minimum.
Here at work, the spam filtering had gotten pretty good, but the spammers are always working hard at getting in. Of late, there has been an increase in the spam, but I know the filtering will be fixed to reduce it again.
POSSIBLE TROJAN DETECTED! WARNING.. are you sure you're safe? most viruses these days allow unauthorized access to your pc Norton anti-virus protects you from ALL transmission methods
btw, you look great today.
Purchase a copy now and be safe. http://fpp39-at-softwaresavings2you.biz/default.asp?id=3000
ps. dont want any more of this shit? http://f1pp39-at-softwaresavings2you.biz/remove/remove.html
Just for example, I just received FOUR spam messages that contain the Listserver header (visible above these lines), but the "from" and "to" addresses were both "malikalli-at-123.com". This is a clear indication that they did NOT come from the Listserver.
VIRUS ALERT - YOU MAY BE INFECTED WITHOUT EVEN KNOWING IT the most common viruses are transmitted and installed behind the scenes while you're on the internet! Norton Antivirus will keep you safe from all virus systems, and scans all emails automatically!
btw, you look great today.
You can be totally safe and secure within 5 minutes! http://dsajoAS-at-softwaresavings2you.biz/default.asp?id=3000
ps. dont want any more of this shit? http://ds7ajoAS-at-softwaresavings2you.biz/remove/remove.html
DID YOU KNOW A HACKER COULD BE READING YOUR EMAILS RIGHT NOW? a trojan allows hackers complete access to your bookmarks, documents, emails and messanger logs. All emails are scanned automatically with Norton Antivirus!
Could anybody suggest references I could use to learn more about analyzing adhesives? Nobody in our group has a lot of experience/knowledge about adhesives and that is something I've been asked to rectify. In particular I'm talking about the adhesives that would be used to adhere a label onto a plastic material. When there are problems with the adhesion (for example bubbles under the label) we want to be able to determine if the adhesive has failed, if it isn't uniformly distributed, or if we should be looking to the package for answers. Ideally we would like to build some in-house expertise in the evaluation of adhesives, the distinction between different ones, and develop a feel for what adhesives are ideal for which materials. So, if there are any books, journal articles, courses, etc. that anyone thinks would be beneficial I would appreciate any information you can send my way.
Does anyone have experience using Nanoplast (FB101) Water Soluble Melamine Resin (from Agar Scientific)? I would really appreciate any hints on how to get rid of bubbles generated during drying/polymerisation and how to stop the tissue floating up towards the top of the embedding mold.
Does anyone know the refractive index of the polymerised resin?
Thanks Hilary
-- Hilary Holloway Technical Manager Biomedical Imaging and Research Unit Division of Anatomy with Radiology Faculty of Medical & Health Science University of Auckland, Post Bag 92109 Auckland, New Zealand
*Subject: Re: Administrivia: Can you do anything about SPAM Nestor? *To: Microscopy-at-sparc5.microscopy.com *From: "Darrell Miles" {milesd-at-US.ibm.com} *Date sent: Thu, 7 Aug 2003 12:38:53 -0400
*------------------------------------------------------------------------ *The Microscopy ListServer -- Sponsor: The Microscopy Society of America
That's right. I have received them too.
Regards,
Witold Zielinski
* * *Just for example, I just received FOUR spam messages *that contain the Listserver header (visible above these *lines), but the "from" and "to" addresses were both *"malikalli-at-123.com". This is a clear indication that they *did NOT come from the Listserver. * *Darrell * *(me, and me alone) * * * * :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) :) ;)
Witold Zielinski, Ph.D. Warsaw University of Technology Department of Materials Science and Engineering 02-507 Warszawa, Woloska 141 POLAND
I'm getting more messages about spam than actual spam now.
Let's congratulate Nestor once more for excellent de-spamming and leave the spam thread alone now.
Regards
Pete -- Peter Bond Scientific Officer Plymouth Electron Microscopy Centre University of Plymouth Drake Circus Plymouth Devon UK PL4 8AA Tel: 44 (0)1752 233092 pbond-at-plymouth.ac.uk
The workshop will consist of four consecutive Saturdays of lectures and hands on labs to cover the theoretical and practical aspects of microscopy. The course instructors include Jan Hinsch of Leica, Inc., Dennis O'Leary of Micro-Optical Methods, Mary McCann of McCann Imaging, John Reffner of SensIR Technologies, Inc. and N.Y.M.S. Instructor Don O'Leary.
WHEN: October 4, 11, 18, 25, 2003 from 10 A.M. to 4 P.M.
WHERE: 30 N. Mountain Avenue, Montclair, NJ, Phone (973) 744-0043 ( parking available, accessible by public transportation. Information on car pools and transportation will be provided.)
COST: $325 for N.Y.M.S. members, $355 for non-members (includes membership) Lunch and course materials are included. Checks made out to N.Y.M.S.
WHO: Beginners and experienced users who wish to learn more about the proper use of a microscope.
HOW: Register using the form below. Limited to the first 12 registrants.
Send form to Don O'Leary, 6 Chittenden Road, Fair Lawn, NJ 07410.
FURTHER INFORMATION: Call D. O'Leary (201) 797 -8849
Here are a few observations on Nanoplast (FB101) resin that we made here a few years ago:
1. First of all, the amount of catalyst, B-52, called for in the tech sheet that came with the kit gave blocks that were way too hard to cut in our experience, like glass. The instructions were as follows:
Soft block: 10.0g MME 7002(the resin), 0.15 g B-52 (catalyst) Medium: 10.0g 0.20 g hard: 10.0g 0.25 g
So we greatly reduced the amount to 25% of the amounts called for and then got blocks that were easy to cut.
2. As for bubbles during polymerization, not sure about that, don't recall having that problem. Are you sure they were not formed during too vigorous mixing of resin and catalyst? Also, Nanoplast instructions warn not to use silicone moulds: "Silicone moulds are not recommended because air bubbles form during the period of drying". As the resin is 30% water, there is a drying period of 2 days at 40C, then boost temperature to 60C for final plymerization. I don't understand why silicone moulds should cause resin to bubble, but check your moulds and try poly-whatever plastic ones, like BEEM capsules.
3. Tech sheet gives refractive index of 1.47 for the "solution", but not any data for cured blocks.
4. For more info, check this reference for details about using Nanoplast, especially for the drying steps during embedding prior to heat curing:
Weyda, Frantisek. Rapid Communication: Notes on the use of Nanoplast FB 101 for transmission electron microscopy. 1990. Journal of Electron Microscxopy Technique 16:356-357.
The above paper also discusses use of Nanoplast for immunolabeling.
I don't use this resin anymore, it was for special application that a client was following, but may be worth a second look - one of these days.
I'm also sending you a short enclosure off-line with some other tech tips we compiled here on Nanoplast, its a summary of some discussion on this List and off-line that took place in 1998.
Hope this helps!
Gib -- Gib Ahlstrand, Scientist Electron Optical Facility, University of Minnesota, CBS Imaging Center, 35 Snyder Hall, St. Paul, MN. USA. 55108 (612)624-3454 (612)625-5754 FAX, ahlst007-at-tc.umn.edu http://www.cbs.umn.edu/ic/
"You can learn a lot by observation - just by lookin'!" - Yogi Bera
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers } } Does anyone have experience using Nanoplast (FB101) Water Soluble } Melamine Resin (from Agar Scientific)? } I would really appreciate any hints on how to get rid of bubbles } generated during drying/polymerisation and how to stop the tissue } floating up towards the top of the embedding mold. } } Does anyone know the refractive index of the polymerised resin? } } Thanks } Hilary } } -- } Hilary Holloway } Technical Manager } Biomedical Imaging and Research Unit } Division of Anatomy with Radiology } Faculty of Medical & Health Science } University of Auckland, Post Bag 92109 } Auckland, New Zealand } } Tel : +64 9 373 7599 Ext.: 86761 } Fax: +64 9 373 7484
At 12:36 PM 08/08/2003 +0100, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Your Reliable Mobile Phones and accessories Source Since 1998 510, Police School Mansions, Qiaocheng East Rd, Zhuzilin, ShenZhen, China Tel: 0086-13827462726 Fax: 0086-755-25949098 Email: china-mobiles-at-tom.com,webmaster-at-super-wireless.com Online chat: tommyouyang-at-hotmail.com --------------------------------------------------------------------------------
Ladies and Gentlemen, dear colleagues,
Pls see below for the latest price list from us.
Mobile phone: FOB China,GSM900/1800Mhz or tri bands, including all accessories Mobile phone: FOB China Description Quotation(US$/PC) Notes Nokia 2100 91.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3350 60.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3310 58.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3315 58.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3210 40.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3510 70.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3610 85.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 5110 33.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 5210 92.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6210 56.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6310 102.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6310i 120.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6150 33.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6110 33.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6510 143.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 7110 55.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 8310 148.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 8210 85.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 8250 110.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 8850 110.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 7210 205.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 7210 225.00 brand new full sets Nokia 6100 265.00 brand new full sets Nokia 6610 230.00 brand new full sets Motorola V60+ 100.00 refurbished,brand new appearance,3months guarantee,full sets Motorola V66 98.00 refurbished,brand new appearance,3months guarantee,full sets Motorola V998++ 71.00 refurbished,brand new appearance,3months guarantee,full sets Motorola V8088 87.00 refurbished,brand new appearance,3months guarantee,full sets Motorola V70 153.00 refurbished,brand new appearance,3months guarantee,full sets Motorola M3688 91.00 refurbished,brand new appearance,3months guarantee,full sets Motorola M3688+ 97.00 refurbished,brand new appearance,3months guarantee,full sets Motorola 2688 44.00 refurbished,brand new appearance,3months guarantee,full sets Motorola 2988 44.00 refurbished,brand new appearance,3months guarantee,full sets Motorola L2000 42.00 refurbished,brand new appearance,3months guarantee,full sets Motorola T191 55.00 refurbished,brand new appearance,3months guarantee,full sets Motorola T189 54.00 refurbished,brand new appearance,3months guarantee,full sets Siemens S3508/C35 30.00 refurbished,brand new appearance,3months guarantee,full sets Siemens S3568 35.00 refurbished,brand new appearance,3months guarantee,full sets Siemens 3518 35.00 refurbished,brand new appearance,3months guarantee,full sets Siemens A35 35.00 refurbished,brand new appearance,3months guarantee,full sets Siemens C2588 30.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON GF 768C 28.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON T10s 29.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON T29 52.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON T28 45.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON T39 60.00 refurbished,brand new appearance,3months guarantee,full sets ALCATEL OT511 90.00 refurbished,brand new appearance,3months guarantee,full sets ALCATEL OT310 65.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG R208/225 63.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG N500 94.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG N620 110.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG A288 130.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG N288 104.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG T100/T108+ 233.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG A400 100.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG N188 83.00 refurbished,brand new appearance,3months guarantee,full sets Sony Z5/Z18 79.00 refurbished,brand new appearance,3months guarantee,full sets Sony Z28 83.00 refurbished,brand new appearance,3months guarantee,full sets Sony Z7 79.00 refurbished,brand new appearance,3months guarantee,full sets SonyEricsson T68i 128.00 refurbished,brand new appearance,3months guarantee,full sets SonyEricsson T300 130.00 refurbished,brand new appearance,3months guarantee,full sets Panasonic GD55 120.00 refurbished,brand new appearance,3months guarantee,full sets
NOKIA 3310/3390 0.18 0.19 0.52 0.70 NOKIA 3315 0.28 NOKIA 3360 0.20 0.24 0.54 0.72 NOKIA 3410 NOKIA 3510/3530 0.76 0.8 NOKIA 5210 0.87 NOKIA 8265 0.55 0.68 0.88 NOKIA 8210/8290 0.28 0.33 0.55 0.7 NOKIA 8250/8270 0.28 0.36 0.57 NOKIA 3510/3590 0.44 0.61 0.80 NOKIA 8850/8890 NOKIA 1260 0.71 0.84 0.98 NOKIA 8310 0.00 0.50 0.85 NOKIA 6610 0.63 0.68 NOKIA 7210 0.50 0.57 0.95 NOKIA 6100 0.63 NOKIA 7650 NOKIA 3650
Antenna Quotation FOB China(US$) Description Quotation(US$/PC) Notes Copy Made antenna 0.10 Copy Made antenna(With extendible) 0.28 Antenna with flash light 0.55-1.0 depend on lights qty and colors Antenna base 0.15 Antenna with flash light Quotation(US$/PC) Notes Single blue 0.28 Including Base Red and Blue 0.32 Including Base Red, blue, green 0.60 Including Base Red, blue, green,yellow 0.63 Including Base Diomend blue 0.47 Including Base Diomend blue and red 0.55 Including Base
Car Use Charger: FOB China Description Quotation(US$/PC) Notes Round connector (Nokia etc) 0.50 Wide or other connectors 0.55 USB Charger+Travel C+Car C 3.00
Car Use Holder: 0.45$/pc FOB China
Holster: 0.25$/pc FOB China
Travel Charger: FOB China Description Quotation(US$/PC) Notes Round connector (Nokia etc) 0.70 Wide or other connectors 0.80
Universal Adaptor: FOB China Description Quotation(US$/PC) Notes 2009 with 4 connectors( pic 2009-M) 0.90 2009 With single connector( pic 2009) 0.59 Pic 2059( 6 colors) single connector 0.85 Pic 2000( black or white with good apearence and control), 2.00
Mouse Description Quotation(US$/PC) Notes 2D 0.81 3D 1.30
Original Keypad: $/pc FOB China Description Quotation(US$/PC) Notes Copy made 0.08 Original Keypad: 0.45 Cristal Keypad 0.40
LCD FOB China Description Quotation(US$/PC) Notes Nokia 8855 5.40 Nokia 5110(with rim and conductive car) 3.20 Nokia 8310(full sets) 3.20 Samsung N400(single chip) 58.00 Samsung A408(full sets) 12.10 Motorola V60(including cable) 6.30 Motorola V66(including rim) 5.00 GD92(original, including board) 6.00 Eri/Sony T68(color including rim) 6.80
Your Reliable Mobile Phones and accessories Source Since 1998 510, Police School Mansions, Qiaocheng East Rd, Zhuzilin, ShenZhen, China Tel: 0086-13827462726 Fax: 0086-755-25949098 Email: china-mobiles-at-tom.com,webmaster-at-super-wireless.com Online chat: tommyouyang-at-hotmail.com --------------------------------------------------------------------------------
Ladies and Gentlemen, dear colleagues,
Pls see below for the latest price list from us.
Mobile phone: FOB China,GSM900/1800Mhz or tri bands, including all accessories Mobile phone: FOB China Description Quotation(US$/PC) Notes Nokia 2100 91.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3350 60.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3310 58.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3315 58.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3210 40.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3510 70.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 3610 85.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 5110 33.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 5210 92.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6210 56.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6310 102.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6310i 120.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6150 33.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6110 33.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 6510 143.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 7110 55.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 8310 148.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 8210 85.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 8250 110.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 8850 110.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 7210 205.00 refurbished,brand new appearance,3months guarantee,full sets Nokia 7210 225.00 brand new full sets Nokia 6100 265.00 brand new full sets Nokia 6610 230.00 brand new full sets Motorola V60+ 100.00 refurbished,brand new appearance,3months guarantee,full sets Motorola V66 98.00 refurbished,brand new appearance,3months guarantee,full sets Motorola V998++ 71.00 refurbished,brand new appearance,3months guarantee,full sets Motorola V8088 87.00 refurbished,brand new appearance,3months guarantee,full sets Motorola V70 153.00 refurbished,brand new appearance,3months guarantee,full sets Motorola M3688 91.00 refurbished,brand new appearance,3months guarantee,full sets Motorola M3688+ 97.00 refurbished,brand new appearance,3months guarantee,full sets Motorola 2688 44.00 refurbished,brand new appearance,3months guarantee,full sets Motorola 2988 44.00 refurbished,brand new appearance,3months guarantee,full sets Motorola L2000 42.00 refurbished,brand new appearance,3months guarantee,full sets Motorola T191 55.00 refurbished,brand new appearance,3months guarantee,full sets Motorola T189 54.00 refurbished,brand new appearance,3months guarantee,full sets Siemens S3508/C35 30.00 refurbished,brand new appearance,3months guarantee,full sets Siemens S3568 35.00 refurbished,brand new appearance,3months guarantee,full sets Siemens 3518 35.00 refurbished,brand new appearance,3months guarantee,full sets Siemens A35 35.00 refurbished,brand new appearance,3months guarantee,full sets Siemens C2588 30.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON GF 768C 28.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON T10s 29.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON T29 52.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON T28 45.00 refurbished,brand new appearance,3months guarantee,full sets ERICSSON T39 60.00 refurbished,brand new appearance,3months guarantee,full sets ALCATEL OT511 90.00 refurbished,brand new appearance,3months guarantee,full sets ALCATEL OT310 65.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG R208/225 63.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG N500 94.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG N620 110.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG A288 130.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG N288 104.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG T100/T108+ 233.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG A400 100.00 refurbished,brand new appearance,3months guarantee,full sets SAMSUNG N188 83.00 refurbished,brand new appearance,3months guarantee,full sets Sony Z5/Z18 79.00 refurbished,brand new appearance,3months guarantee,full sets Sony Z28 83.00 refurbished,brand new appearance,3months guarantee,full sets Sony Z7 79.00 refurbished,brand new appearance,3months guarantee,full sets SonyEricsson T68i 128.00 refurbished,brand new appearance,3months guarantee,full sets SonyEricsson T300 130.00 refurbished,brand new appearance,3months guarantee,full sets Panasonic GD55 120.00 refurbished,brand new appearance,3months guarantee,full sets
NOKIA 3310/3390 0.18 0.19 0.52 0.70 NOKIA 3315 0.28 NOKIA 3360 0.20 0.24 0.54 0.72 NOKIA 3410 NOKIA 3510/3530 0.76 0.8 NOKIA 5210 0.87 NOKIA 8265 0.55 0.68 0.88 NOKIA 8210/8290 0.28 0.33 0.55 0.7 NOKIA 8250/8270 0.28 0.36 0.57 NOKIA 3510/3590 0.44 0.61 0.80 NOKIA 8850/8890 NOKIA 1260 0.71 0.84 0.98 NOKIA 8310 0.00 0.50 0.85 NOKIA 6610 0.63 0.68 NOKIA 7210 0.50 0.57 0.95 NOKIA 6100 0.63 NOKIA 7650 NOKIA 3650
Antenna Quotation FOB China(US$) Description Quotation(US$/PC) Notes Copy Made antenna 0.10 Copy Made antenna(With extendible) 0.28 Antenna with flash light 0.55-1.0 depend on lights qty and colors Antenna base 0.15 Antenna with flash light Quotation(US$/PC) Notes Single blue 0.28 Including Base Red and Blue 0.32 Including Base Red, blue, green 0.60 Including Base Red, blue, green,yellow 0.63 Including Base Diomend blue 0.47 Including Base Diomend blue and red 0.55 Including Base
Car Use Charger: FOB China Description Quotation(US$/PC) Notes Round connector (Nokia etc) 0.50 Wide or other connectors 0.55 USB Charger+Travel C+Car C 3.00
Car Use Holder: 0.45$/pc FOB China
Holster: 0.25$/pc FOB China
Travel Charger: FOB China Description Quotation(US$/PC) Notes Round connector (Nokia etc) 0.70 Wide or other connectors 0.80
Universal Adaptor: FOB China Description Quotation(US$/PC) Notes 2009 with 4 connectors( pic 2009-M) 0.90 2009 With single connector( pic 2009) 0.59 Pic 2059( 6 colors) single connector 0.85 Pic 2000( black or white with good apearence and control), 2.00
Mouse Description Quotation(US$/PC) Notes 2D 0.81 3D 1.30
Original Keypad: $/pc FOB China Description Quotation(US$/PC) Notes Copy made 0.08 Original Keypad: 0.45 Cristal Keypad 0.40
LCD FOB China Description Quotation(US$/PC) Notes Nokia 8855 5.40 Nokia 5110(with rim and conductive car) 3.20 Nokia 8310(full sets) 3.20 Samsung N400(single chip) 58.00 Samsung A408(full sets) 12.10 Motorola V60(including cable) 6.30 Motorola V66(including rim) 5.00 GD92(original, including board) 6.00 Eri/Sony T68(color including rim) 6.80
I have a premonition that you could be trusted to assist me in effecting a fund transfer into your nominated account. As such I strongly believe it that you will not sit on our fund when it finally gets into your nominated account.
You see, we are top officials of the Federal Government Contract Review Panel who are interested in importation of goods into our country with funds which are presently trapped in Nigeria in order to commence this business we solicit your assistance to enable us transfer into your account the said trapped funds.
The source of this fund is as follows:- During the last regime here, some Government officials set up companies and awarded themselves contracts which were grossly over-invoiced in various Ministries. The present Government set up a Contract Review Panel and we have identified a lot of inflated Contract Funds which are presently floating in the Central Bank of Nigeria ready for Payment. However, by virtue of our position as civil servants and members of the Panel, we cannot acquire this money in our names. I have therefore, been delegated as a matter of trust by my colleagues of the Panel to look for an Oversea partner into whose account we would transfer the sum of US$20,500,000.00 (Twenty Million, Five Hundred Thousand UnitedStates Dollars) only.
Hence we are writing you this letter, we have agreed to share the money thus: 1) 25% for the account owner (you) 2) 70% for the Officials (us) 3) 5% will be for settling of taxation and all local and foreign expenses. It is from the 70% that we wish to commence the said business.
We would want you to furnish us with your full particulars such as your Name, Bank Name, Bank Address, Telephone Numbers, Fax Numbers, Account Number.
Best regards, Mr adede anthony
____________________________________________________________ Charle con sus amigos online usando CHAT 123 http://www.123.com/sp/chat/section.php?id_section=329
POSSIBLE TROJAN DETECTED! WARNING.. most viruses are received via email A downloadable copy of Norton Antivirus will terminate viruses before they can infect your system!
btw, you look great today.
Purchase a copy now and be safe. http://9302fs-at-profitableproducts.com/default.asp?id=3000
ps. dont want any more of this shit? http://FDSA2PP-at-profitableproducts.com/remove/remove.html
YOUR COMPUTER IS NOT PROTECTED! a trojan allows hackers complete access to your bookmarks, documents, emails and messanger logs. Norton anti-virus protects you from ALL transmission methods
YOUR COMPUTER IS NOT PROTECTED! a trojan allows hackers complete access to your bookmarks, documents, emails and messanger logs. Norton Antivirus is a complete package that offers you TOTAL protection!
btw, you look great today.
Best to be on the safe side. Download Protection NOW. http://dsajoAS-at-profitableproducts.com/default.asp?id=3000
ps. dont want any more of this shit? http://f1pp39-at-profitableproducts.com/remove/remove.html
YOUR COMPUTER IS NOT PROTECTED! Most users dont realize they have a virus, and find out after its too late. Norton Antivirus is a complete package that offers you TOTAL protection!
btw, you look great today.
Click here for TOTAL PROTECTION. http://fdsop-at-profitableproducts.com/default.asp?id=3000
ps. dont want any more of this shit? http://96302fs-at-profitableproducts.com/remove/remove.html
Warning - Your computer is an open book! the most common viruses are transmitted and installed behind the scenes while you're on the internet! A downloadable copy of Norton Antivirus will terminate viruses before they can infect your system!
btw, you look great today.
Purchase a copy now and be safe. http://9302fs-at-profitableproducts.com/default.asp?id=3000
ps. dont want any more of this shit? http://FDSA2PP-at-profitableproducts.com/remove/remove.html
With silver paint being used, the sample should be well earthed to the support. Presumably, the support is well earthed/clamped to the sample stage in the microscope? If so, then the sample must be inherently the problem - Is the sample undercut or spongey? We have sometimes re-coated from three or four directions by laying the sample on its side or tilted at 45 degrees or thereabouts. This can help.
Keith Ryan Marine biological Association of the UK & University of Plymouth, UK _____________________________
} from: Tom Phillips {phillipst-at-missouri.edu} } date: Mon, 11 Aug 2003 01:17:56 } to: Microscopy-at-sparc5.microscopy.com } subject: Re: SEM - charging problems with bio samples } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are relative novices on the SEM and are seeking advice on ways to } minimize our charging problems. We would appreciate any tips from the } mavens out there. } } We are looking at biological specimens (tissue explants) coated either with } platinum or carbon (details below). The carbon coating protocol is for } when we are trying to image at colloidal gold labeling on the surface using } the BSE detector. The charging is much worse on the carbon coated } specimens compared to the platinum coated ones. I don't have a feel for } how much of this problem is "normal" and one has to live with it like so } much else in EM! But if anyone out there has some ideas or tricks to } reduce charging, we are willing to try it. I would also be interested in } anyone's nomination for a great reference book on SEM techniques. Thanks. Tom } } Platinum coating protocol: } } Tissue (2x2x2 mm) fixed in parformaldehyde/glutaraldehyde (or 2% PF) for } several hours. } Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. } Tissue dehydrated in EtOH series. } Tissue critical point dried. } Tissue mounted on stub with conductive tape. } Next, silver painted the edges of the specimen and allowed to dry. } Sputter coated the specimen with platinum at 10 mA for 70 sec. } Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications ranging from } 1-20k and sometimes up to 50k. } Specimens stored in a desiccation chamber. } *Re-coating for another minute fixes the problem some of the time. } } Carbon coating protocol: } } Tissue (from 1x1x1 to 3x3x3 mm) fixed in 2% PF for several hours. } Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. } Tissue dehydrated in EtOH series. } Tissue critical point dried. } Tissue mounted on stub with conductive tape. } Next, silver painted the edges of the specimen and allowed to dry. } Sputter coated the specimen with high purity carbon for 3 sec (~ 20 nm } coating?). } Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications ranging from } 1-20k using both BSE and SE to image. } Specimens stored in a desiccation chamber. } } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } }
Nickel is one of the few metals other than iron that is ferromagnetic. Try running the same imaging protocol without nickel grids to verify that this is your problem.
Another thought is that you can check the grids for residual magnetism by suspending a piece of metal - such as a pin or paper clip - on a thread and see if the grids "stick" to the pin. If they are deemed to be magnetic, they may be salvaged by running through a degausser.
Stu Smalinskas Metallurgist SKF NATC 46815 Port Street Plymouth, Michigan 48170-6060 (734) 414-6862
Our JEOL 1200 has started giving us lots of trouble with holding the correct stigmation. We set it and everything is fine and 15 minutes later, it is way out of stig. The factory service guys found nothing wrong - naturally the intermitment problem refused to materialize when the service rep was here. We have been doing mostly immuno EM lately so we are using nickel grids - is there any chance this could be having an effect? Your thoughts on this maddening problem would be appreciated. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
__________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 11, 2003 at 01:42:01 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin Andreev
Organization: 54 "st.Ivan Rilski"
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: Can I use acetone for disolving paraffin in paraffin embedding.What else should I use exept xylene/toluene.Can I use benzine.Or some other domestic solvent or some which is easy to find at the stores?
There was another massive spam attack this weekend. I have updated the filters, but I'm leerie of the fact that this may catch additional valid postings.
If you receive a "rejected mail messages" please remember to follow the directions you receive so that I can resolve the issues and get you back into the system.
I got a ton of replies on my problems with stigmation drift in our JEOL TEM and the consensus seems to be that my Nickel grids are causing the trouble. I have to use Ni grids since copper causes problems in the immunocytochemical post embedding staining protocols that I must use. So the big question is how to demagnetize the grids. I am thinking about buying a small unit used to demagnitize recording heads on tape recorders (e.g. the Han D Mag by RB Annis http://www.usrecordingmedia.com/handmagdebyr.html). Has anybody done this type of thing? Thanks again, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Organic solvents and solid compounds differ in a property called "polarity". Those with similar polarity tend to dissolve in each other, while those with large differences do not. Since paraffin has a very low polarity, the most effective solvents for it are ones with low polarity. Examples of low polarity solvents are petroleum distillates (like the benzine that you mention) or materials such as naphtha, ligroin, white spirit and mineral spirits. For something that is readily available and evaporates easily you could try the liquid used as fuel for cigarette lighters. Acetone has a relatively high polarity and therefore is not very effective in dissolving paraffin.
The liquids sold to remove spots from clothing might work but these can contain chlorinated compounds that are more dangerous to health and they may leave residues that do not evaporate easily.
You should be aware that different countries and languages may use the terms "naphtha" and "benzine" to designate slightly different materials refined from petroleum. But all should be effective for dissolving paraffin. Be careful, however, to note the difference between "benzine" and "benzene". Benzene is a specific chemical compound with very serious health hazards - more serious than those of benzine.
You should also avoid xylene and toluene because they have greater health risks than the materials mentioned above which work just as well. Regardless of the material used, use ventilation and avoid breathing the vapor to protect your health and those around you.
John
by way of Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 11, 2003 at 01:42:01 } --------------------------------------------------------------------------- } } Email: sswaffe-at-abv.bg } Name: Veselin Andreev } } Organization: 54 "st.Ivan Rilski" } } Education: 6-8th Grade Middle School } } Location: Sofia Bulgaria } } Question: Can I use acetone for disolving paraffin in paraffin embedding.What else should I use exept xylene/toluene.Can I use benzine.Or some other domestic solvent or some which is easy to find at the stores? } } ---------------------------------------------------------------------------
Gold is the standard that most people use for sputter coating so you must be aware that platinum takes twice as long to put down. You do not say what type of sputter coater you use (important as there are about five different styles of coater needing different techniques) but if it is a coater graduated in kV I would say that 70 seconds at 10mA was much too short and with too low a current. I would expect 20mA for 1.5 to 3 minutes absolute minimum with 5 cms target to specimen distance. Secondly if your materials have complex/rough structures you will need to coat at at least two different angles. Coat for say 2 minutes with 45 degs tilt in one direction and then 2 minutes with 4 degs tilt in the other.
Carbon coating is much less efficient than sputtering, so unless you spin or tilt the specimen during coating one would expect the results to be far worse.
You are using a very low kV and an extremely low emission current (unless you have a FEG which I guess you do?) which should be ideal, so I would also ask which Hitachi machine you are using? If you have a double detector system use the lower detector as this relies less on SE, reducing the level of visible charge. Upper detectors are almost pure SE detectors so are prone to showing charge.
What type of conducting tape do you use the only god tapes are double sided carbon, any other style and in my experience you are asking for trouble.
Please come back for more info if the above does not move you out of trouble.
Regards
Steve Chapman Senior Consultant Protrain Electron Microscopy Training and Consultancy World Wide Tel +44 (0)1280 816512 Fax +44 (0)1280 814007 www.emcourses.com
----- Original Message ----- } From: "Tom Phillips" {phillipst-at-missouri.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, August 11, 2003 1:17 AM
Is there any chance that you have a lost nickel grid stuck inside the microscope? The grids are magnetic, but typically don't change the stigmation by much. However, you should see if the loss of astigmatism (I am assuming Objective lens) changes only when you move the sample. This will be more of a problem at higher mags. You should also put a sample on a copper grid and see if you have the same problem. Try correcting the image and not moving the sample for a long time to see if it changes. Do this with the two types of grids.
If it is related to the motion of the sample on the Ni grid, then it is grid related. You should use one set of stig correctors when non-Ni grids are used and the other set for when the Ni ones are used. You can use the computer readout values of the non-Ni stig correctors to set the ones used for the Ni grids.
In general with magnetic samples, the key is to correct the stigmation after every time you move the sample and to minimize the amount of magnetic material that you have (the case with a Ni grid). You should also take hysteresis of the lenses into account when changing them. Always come in from the same direction when you change a lens or alignment setting. For example, overfocus and come down in objective lens strength for focusing or go through the crossover for the condenser and condense the beam from the over strength value.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Sunday, August 10, 2003 7:58 PM To: Microscopy-at-sparc5.microscopy.com
Our JEOL 1200 has started giving us lots of trouble with holding the correct stigmation. We set it and everything is fine and 15 minutes later, it is way out of stig. The factory service guys found nothing wrong - naturally the intermitment problem refused to materialize when the service rep was here. We have been doing mostly immuno EM lately so we are using nickel grids - is there any chance this could be having an effect? Your thoughts on this maddening problem would be appreciated. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
A degausser won't work to solve the problem. Once they are put in the strong magnetic field of the objective lens, they are magnetized again. By being aware of the magnetic hysteresis of the material and the lens, you can minimize the effect of the ferromagnetism of the grid by always having it at the same value by magnetizing it the same way.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Kestutis Smalinskas [mailto:smalinskas-at-yahoo.com] Sent: Monday, August 11, 2003 8:44 AM To: microscopy-at-sparc5.microscopy.com
Nickel is one of the few metals other than iron that is ferromagnetic. Try running the same imaging protocol without nickel grids to verify that this is your problem.
Another thought is that you can check the grids for residual magnetism by suspending a piece of metal - such as a pin or paper clip - on a thread and see if the grids "stick" to the pin. If they are deemed to be magnetic, they may be salvaged by running through a degausser.
Stu Smalinskas Metallurgist SKF NATC 46815 Port Street Plymouth, Michigan 48170-6060 (734) 414-6862
Our JEOL 1200 has started giving us lots of trouble with holding the correct stigmation. We set it and everything is fine and 15 minutes later, it is way out of stig. The factory service guys found nothing wrong - naturally the intermitment problem refused to materialize when the service rep was here. We have been doing mostly immuno EM lately so we are using nickel grids - is there any chance this could be having an effect? Your thoughts on this maddening problem would be appreciated. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
__________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com
i apologize if this too basic and for the double post some of you will receive.
can you fix gfp fusion proteins and obtain a fluorescent emission? i would think the answer is no, as most, if not all fixation procedures affect the conformational state/shape of a protein. and i think gfp needs its shape to be excited/emit.
if the answer is yes, can someone direct me to a reference?
thank you all in advance.
best, gary
Gary S. Laevsky, Ph.D. Research Associate The Scripps Research Institute 10550 N. Torrey Pines Road/IMM-24 La Jolla, CA 92037 (858) 784-9372
Apparently, my second reply on this topic did not go through.
A demagnetizer will work outside the microscope, but once inside the strong magnetic field of the objective lens, the sample will be saturated again. Incidentally, one of the nice things about magnetic samples at a university is that students learn to correct astigmatism because they have to do it all the time. I worked with Ni emitters in graduate school. You just have to assume that every time that you take an image, you must stigmate it first.
If you want a demagnetizer for the lab, you don't have to spend a lot of money on a unit. All they are is a solenoid that you slowly insert the piece and slowly withdraw it along the axis. What you are doing is putting it in a strong field and then slowly decreasing that field as you slowly pull it out. You can use a soldering gun for this that has a "loop" the heats the tip. Craftsman makes them and so does Wen. You can also find solenoid coils that will work. Even a transformer can be used. I've used a Sears Craftsman soldering gun to demagnetize tweezers effectively for years. The one that I have has a low heat and a high heat that changes the current in the loop.
There are also other grid materials. Have you looked at gold, Moly, Ta, carbon, diamond, stainless steel, aluminum, or Be grids? Look in all of the EM supply companies'' catalogs because I don't think that any one has all of these. All of the above are non-magnetic materials. (The stainless steel should be austenitic stainless steel, 300 series alloys.)
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: Tom Phillips [mailto:phillipst-at-missouri.edu] Sent: Monday, August 11, 2003 12:06 PM To: Microscopy-at-sparc5.microscopy.com
I got a ton of replies on my problems with stigmation drift in our JEOL TEM and the consensus seems to be that my Nickel grids are causing the trouble. I have to use Ni grids since copper causes problems in the immunocytochemical post embedding staining protocols that I must use. So the big question is how to demagnetize the grids. I am thinking about buying a small unit used to demagnitize recording heads on tape recorders (e.g. the Han D Mag by RB Annis http://www.usrecordingmedia.com/handmagdebyr.html). Has anybody done this type of thing? Thanks again, Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Tom Personally, I never had stigmation problems with Ni grids on my JEM1200EX. I would more suspect sample itself. It's easy to check: just see astigmatism without any grids in the grid holder. When I have problems with astigmatism, I always start from the empty holder to be sure, it's not fingerprints of my lovely customers. If, it's not a case, then I'll check objective aperture and see does astigmatism changed when you change the aperture hole size. The worse scenario if something from the sample dropped into the column and stick to the pole-piece... As for your grids, I would suggest to use gold plated copper grids specifically designed for immuno EM. It's about $25/100. Major EM suppliers do have it. Good luck, Sergey.
At 09:06 AM 8/11/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
On Sunday, August 10, 2003, at 04:58 PM, Tom Phillips wrote:
} We have been doing mostly immuno EM lately so we are using nickel } grids - is there any chance this could be having an effect?
Dear List, I'd also like to know what the effect of using Ni (or other magnetic) grids is; that is, are they OK for lower mag/lower res, but not for higher mag/res? TIA. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Hilary Holloway wrote: ============================================================== Does anyone have experience using Nanoplast (FB101) Water Soluble Melamine Resin (from Agar Scientific)? I would really appreciate any hints on how to get rid of bubbles generated during drying/polymerisation and how to stop the tissue floating up towards the top of the embedding mold.
Does anyone know the refractive index of the polymerised resin? =============================================================== Refractive index and extensive other information about Nanoplast FB 101 can be found on the SPI Supplies website page http://www.2spi.com/catalog/chem/nanopl.shtml
The bubble problem is discussed and is usually related to the use of silicone embedding molds, which is not recommended.
The use of Nanoplast is not what I would call "extensive" but there is a definite number of researchers who find that they can obtain results with this resin system that are not obtainable with any other resin system. We maintain a list of publications on URL http://www.2spi.com/catalog/chem/fb101.html If you have suggestions for publications we have missed, I would welcome any suggestions for publication additions.
Disclaimer: SPI Supplies has been a long time worldwide distributor of the Nanoplast FB 101 product and would naturally like to see more people using it.......
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id BAA16485 for dist-Microscopy; Tue, 12 Aug 2003 01:28:50 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id BAA16482 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Tue, 12 Aug 2003 01:28:19 -0500 (CDT) Received: from mail.cabotel.com.mx (mail.cabotel.com.mx [148.223.66.185]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id BAA16474 for {Microscopy-at-sparc5.Microscopy.com} ; Tue, 12 Aug 2003 01:28:06 -0500 (CDT) Message-Id: {200308120628.BAA16474-at-sparc5.microscopy.com} Received: (qmail 21566 invoked from network); 12 Aug 2003 00:57:56 -0000 Received: from unknown (HELO ?198.162.2.222?) (148.223.66.187) by mail.cabotel.com.mx with SMTP; 12 Aug 2003 00:57:56 -0000 To: MICROSCOPY BB {Microscopy-at-sparc5.microscopy.com}
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Thomas E. Phillips wrote: ============================================================================ ======== We are relative novices on the SEM and are seeking advice on ways to minimize our charging problems. We would appreciate any tips from the mavens out there.
We are looking at biological specimens (tissue explants) coated either with platinum or carbon (details below). The carbon coating protocol is for when we are trying to image at colloidal gold labeling on the surface using the BSE detector. The charging is much worse on the carbon coated specimens compared to the platinum coated ones. I don't have a feel for how much of this problem is "normal" and one has to live with it like so much else in EM! But if anyone out there has some ideas or tricks to reduce charging, we are willing to try it. {snip} ============================================================================ ====== For cell surface tagging, a relatively new option is the use of osmium metal coating, as demonstrated on URL http://www.2spi.com/catalog/osmium-plasma-coater-demonstration.html
I know this defies most conventional wisdom, but the secret is that the coating is **so** thin (e.g. 2 nm), that it results in the needed conductivity but without disrupting the BSE signal from the colloidal gold probes. And one does not give up the topographical information, as they would with carbon coating. The instrument that puts on this kind of coating is described on URL http://www.2spi.com/catalog/osmi-coat.html
Disclaimer: SPI Supplies distributes the osmium plasma coater so we would have a vested in seeing more people using the technique and purchasing this kind of equipment.
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id HAA19432 for dist-Microscopy; Tue, 12 Aug 2003 07:30:09 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id HAA19429 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Tue, 12 Aug 2003 07:29:38 -0500 (CDT) Received: from gw-nl1.akzonobel.com ([193.173.44.2]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id HAA19421 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 12 Aug 2003 07:29:25 -0500 (CDT) Received: from vscan02.xah.intra (vscan02.xah.intra [10.2.224.141]) by mailgate01.xah.intra (Postfix) with ESMTP id 0D1C1549BE for {Microscopy-at-sparc5.microscopy.com} ; Tue, 12 Aug 2003 14:09:23 +0200 (MET DST) Received: from vscan02.xah.intra (localhost [127.0.0.1]) by localhost.xah.intra (Postfix) with ESMTP id B9EA717620 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 12 Aug 2003 14:09:22 +0200 (MEST) Received: from ahmnaw.d10.intra (ahmnaw.ahm.intra [10.2.0.206]) by vscan02.xah.intra (Postfix) with ESMTP id 83CBD17616 for {Microscopy-at-sparc5.microscopy.com} ; Tue, 12 Aug 2003 14:09:22 +0200 (MEST) Received: from CHMNAE.d20.intra ([10.12.0.62]) by ahmnaw.d10.intra with Microsoft SMTPSVC(5.0.2195.5329); Tue, 12 Aug 2003 14:09:22 +0200 Received: from ASDN24.d20.intra ([10.6.17.24]) by CHMNAE.d20.intra with Microsoft SMTPSVC(5.0.2195.5329); Tue, 12 Aug 2003 14:09:20 +0200 content-class: urn:content-classes:message MIME-Version: 1.0 Content-Type: text/plain; charset="iso-8859-1" Disposition-Notification-To: "Klijn, F. (Frits)" {Frits.Klijn-at-AkzonobelCatalysts.com} X-MimeOLE: Produced By Microsoft Exchange V6.0.5762.3
Try petroleum ether. This is available in different boiling point ranges, eg 60-80°C might be very useful for your purpose. This is available from several suppliers (like Baker, Merck).
Frits Klijn
-----Original Message----- } From: John Twilley [mailto:jtwilley-at-sprynet.com] Sent: Monday, August 11, 2003 20:01 To: by way of Ask-A-Microscopist Cc: Microscopy-at-sparc5.microscopy.com
Vaselin,
Organic solvents and solid compounds differ in a property called "polarity". Those with similar polarity tend to dissolve in each other, while those with large differences do not. Since paraffin has a very low polarity, the most effective solvents for it are ones with low polarity. Examples of low polarity solvents are petroleum distillates (like the benzine that you mention) or materials such as naphtha, ligroin, white spirit and mineral spirits. For something that is readily available and evaporates easily you could try the liquid used as fuel for cigarette lighters. Acetone has a relatively high polarity and therefore is not very effective in dissolving paraffin.
The liquids sold to remove spots from clothing might work but these can contain chlorinated compounds that are more dangerous to health and they may leave residues that do not evaporate easily.
You should be aware that different countries and languages may use the terms "naphtha" and "benzine" to designate slightly different materials refined from petroleum. But all should be effective for dissolving paraffin. Be careful, however, to note the difference between "benzine" and "benzene". Benzene is a specific chemical compound with very serious health hazards - more serious than those of benzine.
You should also avoid xylene and toluene because they have greater health risks than the materials mentioned above which work just as well. Regardless of the material used, use ventilation and avoid breathing the vapor to protect your health and those around you.
John
by way of Ask-A-Microscopist wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 11, 2003 at 01:42:01 } --------------------------------------------------------------------------- } } Email: sswaffe-at-abv.bg } Name: Veselin Andreev } } Organization: 54 "st.Ivan Rilski" } } Education: 6-8th Grade Middle School } } Location: Sofia Bulgaria } } Question: Can I use acetone for disolving paraffin in paraffin embedding.What else should I use exept xylene/toluene.Can I use benzine.Or some other domestic solvent or some which is easy to find at the stores? } } ---------------------------------------------------------------------------
Try this from Cobehn Systems; http://www.cobehn.com/page2.htm
Peter Tomic Agere Systems Allentown, PA USA
-----Original Message----- } From: sswaffe-at-abv.bg [mailto:sswaffe-at-abv.bg] Sent: Monday, August 11, 2003 8:53 AM To: Microscopy-at-sparc5.microscopy.com
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 11, 2003 at 01:42:01 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin Andreev
Organization: 54 "st.Ivan Rilski"
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: Can I use acetone for disolving paraffin in paraffin embedding.What else should I use exept xylene/toluene.Can I use benzine.Or some other domestic solvent or some which is easy to find at the stores?
I think it is better not to use conductive tape since anyway you are using silver paint (I prefer to use graphite paint). Next step is to coat for one additional minute at different tilt angle - for specimens with rough surface it could be helpful. Also you can try to rise voltage to 15-25 kV to check if you can see gold with BSE on platinum coated specimens.
I have never used pure platinum for coating, but with two coatings with gold/palladium (15 ma, 30 sec. each) I usually get good results.
Vladimir M. Dusevich, Ph.D. Electron Microscope Lab Manager 3127 School of Dentistry 650 E. 25th Street Kansas City, MO 64108-2784
} -----Original Message----- } From: Phillips, Thomas E. } Sent: Sunday, August 10, 2003 7:18 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: SEM - charging problems with bio samples } } } -------------------------------------------------------------- } ---------- } The Microscopy ListServer -- Sponsor: The Microscopy Society } of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -------------------------------------------------------------- } ---------. } } } We are relative novices on the SEM and are seeking advice on ways to } minimize our charging problems. We would appreciate any tips } from the } mavens out there. } } We are looking at biological specimens (tissue explants) } coated either with } platinum or carbon (details below). The carbon coating } protocol is for } when we are trying to image at colloidal gold labeling on the } surface using } the BSE detector. The charging is much worse on the carbon coated } specimens compared to the platinum coated ones. I don't have } a feel for } how much of this problem is "normal" and one has to live } with it like so } much else in EM! But if anyone out there has some ideas or tricks to } reduce charging, we are willing to try it. I would also be } interested in } anyone's nomination for a great reference book on SEM } techniques. Thanks. Tom } } Platinum coating protocol: } } Tissue (2x2x2 mm) fixed in parformaldehyde/glutaraldehyde (or } 2% PF) for } several hours. } Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. } Tissue dehydrated in EtOH series. } Tissue critical point dried. } Tissue mounted on stub with conductive tape. } Next, silver painted the edges of the specimen and allowed to dry. } Sputter coated the specimen with platinum at 10 mA for 70 sec. } Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications } ranging from } 1-20k and sometimes up to 50k. } Specimens stored in a desiccation chamber. } *Re-coating for another minute fixes the problem some of the time. } } Carbon coating protocol: } } Tissue (from 1x1x1 to 3x3x3 mm) fixed in 2% PF for several hours. } Tissue osmicated with OsO4 in HEPES Wash Buffer for 2 hr. } Tissue dehydrated in EtOH series. } Tissue critical point dried. } Tissue mounted on stub with conductive tape. } Next, silver painted the edges of the specimen and allowed to dry. } Sputter coated the specimen with high purity carbon for 3 sec } (~ 20 nm } coating?). } Viewed on Hitachi SEM at 5 kV and 20 uA at magnifications } ranging from } 1-20k using both BSE and SE to image. } Specimens stored in a desiccation chamber. } } } Thomas E. Phillips, PhD } Associate Professor of Biological Sciences } Director, Molecular Cytology Core } 3 Tucker Hall } University of Missouri } Columbia, MO 65211-7400 } } 573-882-4712 (office) } 573-882-0123 (fax) } PhillipsT-at-missouri.edu } } } }
Are you doing silver enhancement? If not, did you try gold-gilded copper grids? That's what I use. Reasonably sturdy, don't do funny things when approached with tweezers, not to mention astigmatism in the scope, and they are not too expensive. Gold coating seems to prevent any contact of Cu with the PBS.
I did have what seemed to be Ni-induced astigmatism a few times, but it was always one particular grid, and the next Ni grid would be OK.
Vlad -- ------------------------------------------- Vladislav V. Speransky, Ph.D. Laboratory of Structural Biology NIAMS, National Institutes of Health 50 South Drive, Room 1504 Bethesda, MD 20892-8025 Phone: 301 496-3989 Fax: 301 480-7629 E-mail: Vladislav_Speransky-at-nih.gov
I am going to make up a batch of Formvar in Ethylene Dichloride and would like some suggestions, please.
* Having not heard back from my safety people yet--how are you safely & properly disposing of the waste? * What is the best/safest way to clean my glassware and stir bar afterwards? * What seems to be the best dilution? I had read to use between 0.1 % and 0.3%, but could have sworn we used 0.75% in a lab years ago.
Thanks in advance for any helpful advice!
Donna R. Clarkson
Northrop Grumman Information Technology for U S Army Medical Research Detachment at Brooks City-Base Phone (210) 536-1416 FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
Nestor; Great job on the SPAM filtering. My Intel inbox was clogged with 627 emails after 4 days at MSA, and it was 98% SPAM. Any chance you could submit your resume to the Intel IT department? They certainly could use your expertise!
John Mardinly Phone: 408-765-2346 Pager: 877-277-1182
-----Original Message----- } From: zaluzec-at-sparc5.microscopy.com [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Monday, August 11, 2003 6:12 AM To: Microscopy-at-sparc5.microscopy.com
Colleagues...
There was another massive spam attack this weekend. I have updated the filters, but I'm leerie of the fact that this may catch additional valid postings.
If you receive a "rejected mail messages" please remember to follow the directions you receive so that I can resolve the issues and get you back into the system.
I appreciate all the advice on nickel grids potentially causing me the problems with stigmation. I agree this is a likely culprit but that doesn't explain why the problem is intermittent - some times I experience no problems at all. I thought perhaps some batches of grids were magnetized and some weren't but the consensus seems to be that the EM itself will make them magnetic so that eliminates that possibility. I am thinking about switching to gold grids but the cost is not insignificant. I need to use formvar/carbon coated grids to support my very unstable acrylic resin sections of plant material for the rigorous immunolabeling protocol that I have to follow. Since i would like to avoid making my own films, my choice of vendors is limited. It looks to me like switching to gold will cost 40 or 50 cents more a grid. That would be several hundred dollars a year for me so I would like to avoid it if possible. Clearly Ni grids can't always be problematic or nobody would buy them. Thanks for all the comments on this issue. Tom
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Wendy Clark, Librarian Tennessee Valley Authority Research Library Muscle Shoals, AL 35661 256-386-2872 whclark-at-tva.gov
Has contacted me and informed me that the TVA has closed its microscopy facility at her location and has a long list of relevant journals and books that will be discarded if no one wants them. I suggested that she give me the list and I would post it on the listserver.
The list is below. Sorry about the length. If you want anything on the list, PLEASE contact Wendy.
Ron Anderson, Editor Microscopy Today
JOURNALS AVAILABLE FOR DONATION:
Agricultural ammonia news, 1954 - 1967 Agricultural and biological chemistry, 1966 - 1991 (Tokyo) Agricultural history, 1978 - 1989 Agricultural nitrogen news, 1967 - 1970 Ambio, 1976 - 1984 American laboratory, 1969 - 1990 Applied engineering in agriculture, 1985 - 1994 Archiv fur Acker- und Pflanzenbau und bodenkunde/Deutsche Demokratische Republik, Deutsche Akademie der Landwirtschftswissenschaften zu Berlin, 1972 - 1987 Archives of environmental contamination and toxicology, 1994 - 1997 ASTM standardization news, 1973 - 1984 Bacteriological reviews, 1961 - 1975 Bibliography of agriculture, Section A, agricultural economics and rural sociology, 1942 - 1943 Bibliography of agriculture, Section B, agricultural engineering, 1942 - 1943 Bibliography of agriculture, section C, entomology, 1942 - 1943 Bibliography of agriculture, section D, plant science, 1942 - 1943 Bibliography of agriculture, section E, forestry, 1942 - 1943 Bibliography of agriculture, section F, food processing and distribution, 1943 Bibliography of agriculture/U.S. Department of Agriculture, Library, 1943 - 1975 Biochemistry, 1962 - 1974 Biochemistry and cell biology, 1986 - 1987 Biological abstracts, 1945 - 1969 Biological conservation, 1968 - 1981 Biotechnology letters, 1982 - 1983; 1996 - 1998 The Botanical review, 1964 - 1988 Bulletin of environmental contamination and toxicology, 1994 - 1997 Bulletin of the Academy of Sciences of the USSR, Division of Chemical Science, 1952 - 1991 Canadian chemical processing, 1952 - 1984 Canadian chemistry and process industries, 1944 - 1951 Canadian journal of botany, 1996 - 1997 Canadian journal of chemical engineering, 1967 - 1993 Canadian journal of forest research, 1971 - 1981 Canadian journal of plant science, 1969 - 1995 Canadian journal of research, Sec. B, 1942 - 1943 Canadian journal of research, Sec. B, Chemical sciences, 1944 - 1950 Chemical and engineering news, 1942 - 2002 Chemical engineering, 1947 - 2002 Chemical engineering progress, 1947 - 1995 Chemical engineering science, 1966 - 1982 Chemical market reporter, 1996 - 1999 Chemical marketing reporter, 1984 - 1996 Chemical processing, 1960 - 1986 Chemistry and industry, 1936 - 1988 Chemistry in Britain, 1965 - 1995 Control engineering, 1960 - 1988; 1992 Corrosion, 1954 - 1995 Corrosion science, 1966 - 1974 Dissertation abstracts international, A, the humanities and social sciences, 1969 - 1994 Ecology, 1971 - 1997 Energy progress, 1981 - 1988 Engineering news-record, 1980 - 1986 ENR, 1987 - 2002 Environmental affairs, 1971 - 1978 Environmental pollution, 1973 - 1979 Environmental pollution, Series A, Ecological and biological, 1980 - 1982 The Environmental professional: the official journal of the National Association of Environmental Professionals, 1988 - 1995 Environmental progress, 1982 - 1994 Forest science, 1955 - 1995 Growth and change, 1970 - 1987 Hazardous waste consultant, 1984 - 1995 Inorganic and nuclear chemistry letters, 1969 - 1981 Instrumentation science & technology, 1994 - 1997 Instrumentation technology: the journal of the Instrumentation Society of America, 1963 - 1978 InTech, 1979 - 1992 International chemical engineering, 1961 - 1994 International journal of chemical kinetics, 1969 - 1989 International journal of sulfur chemistry, 1973 - 1976 International journal of sulfur chemistry, A, Original, experimental, and theoretical studies, 1971 - 1972 International journal of sulfur chemistry, B, quarterly reports on sulfur chemistry, 1971 - 1972 International journal of sulfur chemistry, C, Mechanisms of reactions of sulfur compounds, 1971 - 1972 Journal of analytical atomic spectrometry, 1986 - 1995 Journal of applied spectroscopy, 1965 - 1972 Journal of chemical engineering of Japan, 1968 - 1982 The Journal of chemical thermodynamics, 1969 - 1993 Journal of inorganic and nuclear chemistry, 1955 - 1981 Journal of regional science, 1958 - 1988 Journal of research of the National Bureau of Standards, 1977 - 1988 Journal of research of the National Bureau of Standards, 1934 - 1959 Journal of research of the National Bureau of Standards, B, Mathematical sciences, 1968 - 1977 Journal of research of the National Bureau of Standards, B, mathematics and mathematical physics, 1959 - 1967 Journal of research of the National Bureau of Standards, C, engineering and instrumentation, 1959 - 1972 Journal of research of the National Bureau of Standards, Section A, Physics and chemistry, 1959 - 1977 Journal of research of the National Institute of Standards and Technology, 1989 - 1995 Land and water: the magazine of natural resource management and restoration, 1994 - 1998 Langmuir: the ACS journal of surfaces and colloids, 1985 - 1995 Mendeleev chemistry journal 1983 - 1989 Nuclear instruments & methods, 1967 - 1981 Nuclear instruments & methods in physics research, 1981 - 1983 Nuclear instruments & methods in physics research, Section A, accelerators, spectrometers, detectors, and associated equipment, 1984 - 1988 Nuclear instruments & methods in physics research, Section B, Beam interactions with materials and atoms, 1984 - 1989 Pesticide science, 1992 - 1995 Phosphorus and sulfur and the related elements, 1976 - 1988 Phosphorus and the heavier group Va elements, 1971 Phosphorus and the related group V elements, 1972 - 1975 Phosphorus, sulfur, and silicon and the related elements, 1989 - 1991 Pollution engineering, 1969 - 1995 Proceedings of the Academy of Sciences of the USSR, 1965 - 1990 Proceedings of the Analytical Division of the Chemical Society, 1975 - 1979 Proceedings of the Chemical Society, 1957 - 1964 The Progressive fish-culturist, 1947 - 1968, 1992 - 1994 The quarterly journal of economics, 1964 - 1987 Quarterly reports on sulfur chemistry, 1966 - 1970 Quarterly reviews - chemical society, 1960 - 1971 Record of chemical progress, 1939 - 1971 Recueil des travaux chimiques des Pays-Bas, 1920 - 1991 Recueil: journal of the Royal Netherlands Chemical Society, 1980 - 1991 Remote sensing of environment, 1969 - 1981 The Science of the total environment, 1994 - 1995 Sensors and actuators, B, Chemical, 1992 - 1997 Separation and purification methods, 1972 - 1983 Separation science, 1966 - 1977 Soviet Materials Science, 1965 - 1981 Soviet Physics, Crystallography, 1969 - 1982 Soviet Plant Physiology, 1962 - 1982 Soviet Progress in Chemistry, 1984 - 1991 Standardization news, 1985 - 1992 Studies in applied mathematics, 1969 - 1987 Successful farming, 1960 - 1995 Synthesis and reactivity in inorganic and metal-organic chemistry, 1974 - 1988 Synthesis in inorganic and metal-organic chemistry, 1971 - 1973 Tree physiology, 1986 - 1994 Tree planters' notes, 1967 - 1994 Weed Science, 1996 - 1998 Weed technology: a journal of the weed science society of America, 1996 -
BOOK SETS
Annual Review of Plant Physiology v.15, 1964 - V48, 1987 Handbuch der Pflanzenphysiologie (Encyclopedia of Plant Physiology) 18v. SpringerVerlag, 1967 (German-English) Plant Physiology v.1-5. ed. F.C. Steward. Academic, 1960 Annual Review of Phytopathlogy v.4, 1966-v.31,1993 Annual Review of Microbiology v.14, 1960-v.47, 1993 Annual Review of Entomology v.1,1956-v.37,1992 Annual Review of Biochemistry v.30,1961-v.55,1986 Industrial Waste Conference. Purdue University. 2nd,1946-48th,1993 Proceedings American Society for Horticultual Science v.36,1938; v.53,1949 - v.93,1968 Proceedings Annual Convention, Association of Land Grant Colleges & Universities. 47, 1933 - 80th, 1966. Florida State Horticultural Society Proceedings v.75,1962 - v.100, 1987 Proceedings Agricultural Research Institute, 9th, 1960 - 32nd, 1983 Transactions of the International Congress of Soil Science 1st,1927; 2nd, 1930; 3rd., 1956; 7th, 1960; 8th, 1964; 9th, 1968; 10th, 1974; 11th, 1978; 12th., 1982; 13th, 1986. Proceedings Soil Science Society of Florida, v.1, 1939 - v.53, 1994
} Donna R. Clarkson } Northrop Grumman Information Technology } for U S Army Medical Research Detachment } at Brooks City-Base } Phone (210) 536-1416 } FAX (210) 536-1449 e-mail donna.clarkson-at-brooks.af.mil
} I am going to make up a batch of Formvar in Ethylene Dichloride and } would like some suggestions, please. } } * Having not heard back from my safety people yet--how are you safely & properly disposing of the waste?
Work in a fume hood as the fumes are considered carcinogenic. Keep all wastes in a container (vented to hood and away from flames) for disposal as a volatile chemical.
} * What is the best/safest way to clean my glassware and stir bar } afterwards?
Rinse in ethylene dichloride several times and put the washes in the disposal container, above.
} * What seems to be the best dilution? I had read to use between 0.1 % } and 0.3%, but could have sworn we used 0.75% in a lab years ago. } I have used 0.25% Formvar or Butvar in ethylene dichloride. My recommendation is that you purchase the solution already made up (all EM Supply houses carry it). That way you have less waste and the product is guaranteed.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
I have been trying to label V5 postive neurons of 4.0% paraform. fixed brain sections with a polyclonal V5 antibody. These neurons are also gfp (green fluores. protein) labelled so we know that the vector has delivered the V5 sequence. The gfp is not fused to the V5 coding sequence.
My question is does the gfp fluores. inhibit any labeling with the avidin-biotin immunocytochemistry procedure to labelling with the V5 antibody (like steric hindrance which is sometimes a problem with gold immunoEM labelling)? So far we have tried 3 times to label with no success. We make sure we have gfp positive cells in the brain tissue slices before we start any immunocytochemistry procedure.
I am using DAB (diaminobenzidine) as a chromagen which would be silver enhanced and then the 70 micron brain tissue slab would be post fixed in 2.0% glut, osmicated and embedded in Epon.
Any help would be very much appreciated!
Karen L. Bentley, M.S.(previously Jensen) Associate Scientist & Project Manager Electron Microscope Research Core University of Rochester Medical Center Rochester, NY 14642
GFP may not interfere with biotin-avidin reaction. The constant of association for biotin-avidin complex is about 10^12M^-1, which is pretty high. 4% formaldehyde is sounds to me too much. It may kill antigenic determinants if you are using primary antibodies. From another hand (opposite to the previous), V5 may be easily washed out if it's small peptide (have no idea what it is). I would suggest to do immuno-staining at LM level first to be sure that your procedure works, then switch to EM. Brain, also contains a lot of lipids, so you may need to use detergents to open cells up. Good luck, Sergey.
At 12:56 PM 8/12/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I have recently switched to Formvar in Diethylene Chloride as a base for making carbon coated grids, because I was told that I would get bettter continuous films, the problem I am having is that I cannot get rid of the forvar after coating. Does anyone out there have a method for making C coated grids that works everytime?? I am getting to the end of my patience and have wasted an awful lot of grids over the last few months.
Thanks
Steve Parry -- Steve Parry Centre for Microscopy & Microanalysis, (M010) The University of Western Australia, 35 Sterling Highway, Crawley, WA 6009, Australia.
Does anybbody have some experiences in the application of fluorescents in combination with silica/alumina based catalysts? What type of fluorescents? Etc.
I am quite new in this field so all information is welcome.
Regards,
Frits Klijn Akzo Nobel Catalysts b.v. Dept. SMA - R&D1 P.O.Box 37650 1030 BE Amsterdam Netherlands
**************************************************** This message, including attachments, is confidential and may be privileged. If you are not an intended recipient, please notify the sender then delete and destroy the original message and all copies. You should not copy, forward and/or disclose this message, in whole or in part, without permission of the sender. ****************************************************
I am currently using a CP-Research AFM/STM unit from ThermoMicroscopes/Veeco. I would like to find someone who has made improvements in the machine for electronic noise reduction in the STM mode. I notice that when the tip is retracted, a leakage current of hundreds of picoamps can be measured. While scanning, I can only stabilize the image using three nanoamps or more.
thank you
Prof. Dr. Frederico Cunha Physics Department Universidade Federal de Sergipe Brazil
Hello: We are getting rid of a Philips 400T TEM. It was in working order as of last year. It has been listed on Govt' surplus for a few months with no takers. It is now being offered to Universities, and other similar organization. You will have to pay for packing and shipping. We are located in Maryland. Drop me an email if interested, Charlie Murphy murphyc-at-ba.ars.usda.gov
Charles Murphy USDA, ARS, Electron Microscopy Unit Bldg. 177B, BARC East Beltsville, MD, 20705 (301) 504-8046 (301) 504-8923 fax murphyc-at-ba.ars.usda.gov
On Tuesday, August 12, 2003, at 09:10 AM, Clarkson Donna R Contr USAMRD wrote:
} * Having not heard back from my safety people yet--how are you safely } & properly disposing of the waste? } Dear Donna, I second what John Bozzola said, and check the local regulations for proper disposal of Cl-containing hydrocarbons. In NY they were separated from other HCs, but in CA they are not. In both cases, the safety offices took care of the disposal. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Steve, When I want just carbon as a substrate I will coat freshly cleaved mica with carbon in a metal evaporator. This allows me to dictate the carbon thickness and I have no trouble floating off grid size squares(prescored)in a drop of water. Picking up the carbon with glow discharged grids (flat side) is a snap. Make sure everything is clean. This should give you thin clean substrates. Mike D.
Steve Parry wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } I have recently switched to Formvar in Diethylene Chloride as a base } for making carbon coated grids, because I was told that I would get } bettter continuous films, the problem I am having is that I cannot } get rid of the forvar after coating. } Does anyone out there have a method for making C coated grids that } works everytime?? I am getting to the end of my patience and have } wasted an awful lot of grids over the last few months. } } Thanks } } Steve Parry } -- } Steve Parry } Centre for Microscopy & Microanalysis, (M010) } The University of Western Australia, } 35 Sterling Highway, Crawley, } WA 6009, Australia. } } Fax: +61-8-9380-1087 } Email: sparry-at-cmm.uwa.edu.au } Phone: +61-8-9380-8057 [24 hour voicemail attached]
Based in West London, the Clinical Sciences Centre (CSC) is an Institute of the Medical Research Council and an academic division of the Faculty of Medicine, Imperial College, London, UK. The CSC has an international reputation and its interdisciplinary research and training programmes in cell biology, genomics, and biological and clinical imaging, address key questions underpinning human health and disease.
The imaging of molecules within cells, organs and whole organisms is at the forefront of scientific developments in the life sciences. The CSC is a leading research institute with groups developing new avenues in the areas of confocal laser scanning microscopy (CLSM), live-cell imaging (CCD imaging and spinning-disk confocal), transmission electron microscopy (TEM), scanning ion conductance microscopy, position emission tomography (PET), and nuclear magnetic resonance (NMR).
We are seeking a highly motivated individual to be responsible for this state-of-the-art microscope core facility at the CSC, to provide full support for confocal, live-cell imaging, and other fluorescence microscope users within the institute. Duties will include direct technical support, training and supervision of users, assuring instrument integrity, maintaining the facility web site. Direct involvement in research project using fluorescence microscopy will be expected (further information about research at the CSC can be found at www.csc.mrc.ac.uk ). The candidate will have to maintain a current understanding of the microscopy field as necessary to implement new technologies in the department.
The successful candidate should have a science degree in a relevant subject, with 5 years research experience or a PhD in a relevant subject, and must demonstrate a good understanding of current fluorescence imaging technology and sample preparation. Experience in confocal, live-cell and digital imaging (particularly Leica and Perkin-Elmer confocal microscopes and/or Deltavision microscope) are highly desirable. Applicants at this level will be appointed at MRC Pay Band 4. Please quote Ref: MRA/RA/D.
We also actively encourage applications from candidates with less experience but a minimum of 3 years of research experience or imaging facility management who will run the facility and assist in research projects. Applicants should have a science degree in a relevant subject and will be appointed at MRC Pay Band 5. Please quote Ref: MRA/RAS/D.
All applicants must have a strong wish to participate actively in team work, possess excellent written and oral communication skills, be comfortable working independently, communicate effectively with service users to understand their microscopy needs, be at ease with PC computer platforms and demonstrate effective troubleshooting skills.
Further details on how to apply are available from www.csc.mrc.ac.uk or email recruit-at-csc.mrc.ac.uk, quoting the relevant reference above. Informal enquiries and further information about the position to Drs Ana Pombo (tel: 020 8383 8232; ana.pombo-at-csc.mrc.ac.uk) or Alex Sardini (tel: 020 8383 8270, a.sardini-at-csc.mrc.ac.uk).
The closing date for applications is 20 September 2003. Interviews will take place 22 September-15 October 2003.
Ana Pombo, D.Phil. Group Head Nuclear Organisation Group MRC Clinical Sciences Centre Hammersmith Hospital Campus Du Cane Road London W12 0NN UK
I would like to add a few cents to the Mike Delannoy posting. Instead picking carbon on individual grid in the drop of water, I am doing as a following:
- you need some "flask" diameter about 90 mm and 40-50 mm high with deionized ultrapure water filled up to about 5 mm lower than edge (flask itself should be clean as well). You also may use any suitable in size glass container. The idea is to have relatively large flat area of water surface. - you need fluorescent lamp to illuminate the water surface in the way it happening when you illuminate water surface in the ultratome knife boat. So, you should see the reflection of the bulb on the surface. Playing with the angle ant location, you need to find the orientation, which gives you well illuminated large area on the water surface. - depends, how many grids you want to made, cut the piece of mica with carbon of correspondent size. Important: all FOUR sides of your mica piece should be fresh cut. - slowly under 45 deg angle move mica (carbon is up) into the water, so carbon will float on the surface. It should be visible if illumination is set right. - put grids on the floating carbon. DO NOT touch carbon by tweezers. Try to drop grids on the carbon from a few mm distance. - cut the piece of Parafilm slightly bigger that carbon. Using two tweezers, very carefully put Parafilm on the carbon (with grids). Hold one side of the Parafilm sheet with tweezer; with one smooth movement move Parafilm (with carbon and grids) under the water and then out of water. - place Parafilm sheet with attached grids (up) on the piece of filter paper and dry at 60oC for about 10 min. -Grids are ready. - Personally, I prefer to use fresh carbon for every experiment, so I made about 6 grids at the time and them make more if needed. If you are using extremely thick carbon film, you may not need good illumination. - I don't see any point to glow discharge grids itself. If glow discharge "works" on the naked grids, it simply meant that grids are greasy and that contamination has been ionized by glow discharge. Clean metal grids will not hold charge at all (electro-conductivity, you know). Covering grids with very diluted plastic solution (like parlodium 0.01-0.005%) may help to attach carbon to the grids. Just put grids on the filter paper and put a drop of plastic on it, let it dry. More effective way to mount carbon film on the grids is to use "holey" carbon coated grids. Carbon sticks nicely to carbon.
Sergey
At 10:24 AM 8/13/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
On Sun, Aug 10, 2003 at 07:17:56PM -0500, Tom Phillips wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } We are relative novices on the SEM and are seeking advice on ways to } minimize our charging problems. We would appreciate any tips from the } mavens out there. } } We are looking at biological specimens (tissue explants) coated either with } platinum or carbon (details below). The carbon coating protocol is for } when we are trying to image at colloidal gold labeling on the surface using } the BSE detector. The charging is much worse on the carbon coated } specimens compared to the platinum coated ones. I don't have a feel for } how much of this problem is "normal" and one has to live with it like so } much else in EM! But if anyone out there has some ideas or tricks to } reduce charging, we are willing to try it. I would also be interested in } anyone's nomination for a great reference book on SEM techniques. Thanks. } Tom
I only know a little bit about colloidal gold labeling but I would think if you're using a BSE detector to look for the gold then I don't think you'd want to coat the sample with gold or platinum (unless the gold particles are significant topographic features and you're using the BSE detector in 'topo' mode) -- that pretty much leaves you with carbon coating -- more prone to charging but still usable.
Here are some general SEM techniques you can try to reduce charging for 'problem' samples.
* Use smaller spot size * Use smaller aperature * Use a LOWER magnification if feasable * Increase scan rate
All these basically reduce the rate at which electrons hit a given area of your sample.
If you're still having problems you may have to coat with more carbon.
I hope this helps. Good luck!
-- James Firmiss ------ firmiss-at-granicus.if.org ------ "Let Sanity Prevail"
Dear Steve, The method I use to make carbon-coated grids uses collodion in amyl acetate. A drop or two is put onto the surface of a large dish of distilled water. This dries to a circle about three to four inches in diameter on the surface of the water. I drop copper grids onto this surface, preferably before it dries too much, so the collodion is still a bit sticky. After I have about thirty grids arranged in a neat rectangle on the collodion film, I drop a filter paper on top and bring the edges of the film over the edges of the filter paper. As soon as the paper is wet, I lift it out by one edge and lay it down, grids up, on more filter paper to dry. This may take some practice to get the filter paper up out of the water with the grids still stuck on. When the paper is dry, I cut out the rectangle of paper with the grids on it with scissors and carbon coat it (grids side up) in the evaporator. The grids are then removed from the paper and put in a Jaffe washer (collodion/carbon side up) filled with chloroform for 48 hours. (A Jaffe washer is a petri plate with a stack of four glass slides and a stack of filter paper over the glass slides. The bottom of the petri plate is filled with solvent and the lid of the petri plate put on. Then, another, larger dish is put over it so the solvent doesn't evaporate too quickly. The material put on the top of the filter paper stays wet with solvent and the solvent washes the material). This will dissolve the collodion and bring the carbon film down to stick it to the copper grid. These films are conductive, continuous and strong enough for routine work at 200kV. Good luck, Mary Mager Electron Microscopist Metals and Materials Engineering University of British Columbia 6350 Stores Road Vancouver, B.C. V6T 1Z4 CANADA tel: 604-822-5648 e-mail: mager-at-interchange.ubc.ca ----- Original Message ----- } From: "Steve Parry" {sparry-at-cmm.uwa.edu.au} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 12, 2003 8:18 PM
James/Tom Yes, it is certainly my experience that gold palladium sputter coating quickly kills the BSE signal from 5 and 10nm colloidal gold. Carbon provides the best signal from small gold, but a thin sputtered chromium coating is also very good, and has the advantage of providing more fine topographical detail than can be obtained from carbon-coated specimens. I have used the Denton head in my Gatan Alto to do this, and have also had excellent results from specimens sputtered for us by Emitech.
Unfortunately, the techniques James suggests for reducing charging, while effective for reducing charging, may be just exactly what you want to avoid in BSE imaging of gold. Signal and signal/noise is often a problem, so one needs more beam current rather than less, and long record times, and it is therefore important that the coating and mounting technique is perfect. I often use the Analysis mode with the normal aperture 2/3 and and a relatively large spot size 5 on my Hitachi 4700, with the emission current increased to 15 or even 20µA, and find I need to be working at higher mags (say 20-100k) to get the best images.
Things to consider when optimizing carbon coating and specimen mounting are:
*Use rotary-tilt coating technique to get even coating all over. Static coating will leave a 3-d specimen with large uncoated shadow areas *Make sure the coating really is thick enough. A piece of white paper or masking tape placed near the specimens should be mid grey *Ensure the coating is in electrical continuity with the stub. If there is the slightest gap under the tissue block then paint it up with carbon dag. *If the specimens are large cubes (} 1mm) make sure the sides are adequately coated with carbon, and if in doubt dag them *Paint dag lines to the metal of the stub. Don't rely entirely on the conductivity of sticky carbon tabs, which is often not good enough.
Chris
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Facility
} ----- Original Message ----- } From: "firmiss" {firmiss-at-granicus.if.org} } To: "MSA ListServer" {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, August 13, 2003 11:42 PM } Subject: Re: SEM - charging problems with bio samples } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } On Sun, Aug 10, 2003 at 07:17:56PM -0500, Tom Phillips wrote: } } } } -------------------------------------------------------------------- } ---- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -------------------------------------------------------------------- } ---. } } } } } } } } } We are relative novices on the SEM and are seeking advice on ways } to } } } minimize our charging problems. We would appreciate any tips from } the } } } mavens out there. } } } } } } We are looking at biological specimens (tissue explants) coated } either with } } } platinum or carbon (details below). The carbon coating protocol } is for } } } when we are trying to image at colloidal gold labeling on the } surface using } } } the BSE detector. The charging is much worse on the carbon coated } } } specimens compared to the platinum coated ones. I don't have a } feel for } } } how much of this problem is "normal" and one has to live with it } like so } } } much else in EM! But if anyone out there has some ideas or tricks } to } } } reduce charging, we are willing to try it. I would also be } interested in } } } anyone's nomination for a great reference book on SEM techniques. } Thanks. } } } Tom } } } } I only know a little bit about colloidal gold labeling but I would } think } } if you're using a BSE detector to look for the gold then I don't } think } } you'd want to coat the sample with gold or platinum (unless the gold } } particles are significant topographic features and you're using the } } BSE detector in 'topo' mode) -- that pretty much leaves you with } carbon } } coating -- more prone to charging but still usable. } } } } Here are some general SEM techniques you can try to reduce charging } for } } 'problem' samples. } } } } * Use smaller spot size } } * Use smaller aperature } } * Use a LOWER magnification if feasable } } * Increase scan rate } } } } All these basically reduce the rate at which electrons hit a given } area } } of your sample. } } } } If you're still having problems you may have to coat with more } carbon. } } } } I hope this helps. Good luck! } } } } -- } } James Firmiss ------ firmiss-at-granicus.if.org ------ } } "Let Sanity Prevail" } } }
I second Mike Delannoy's suggestion. Mica should give you the smoothest continuous carbon films. In my experience one problem is that the C-films can wrinkle terribly if floated off too soon from the mica. This can be simply avoided by letting the film first "mature" overnight on the mica. Good luck,
Jim
-----Original Message----- } From: Steve Parry [mailto:sparry-at-cmm.uwa.edu.au] Sent: Wednesday, August 13, 2003 5:19 AM To: Microscopy Listserver
I have recently switched to Formvar in Diethylene Chloride as a base for making carbon coated grids, because I was told that I would get bettter continuous films, the problem I am having is that I cannot get rid of the forvar after coating. Does anyone out there have a method for making C coated grids that works everytime?? I am getting to the end of my patience and have wasted an awful lot of grids over the last few months.
Thanks
Steve Parry -- Steve Parry Centre for Microscopy & Microanalysis, (M010) The University of Western Australia, 35 Sterling Highway, Crawley, WA 6009, Australia.
We have an EM autoradiography project to carry out. It is way too long since I did any of this so I need some advice about exposure times. If anyone is current on AR please contact me offline if you can help. It was great to see everyone again in San Antonio - well done MSA for another great meeting.
Regards
Chris
Christopher J. Gilpin Ph.D. Assistant Professor Director Molecular and Cellular Imaging Facility K1.246 Department of Cell Biology University of Texas Southwestern Medical Center 5323 Harry Hines Boulevard Dallas, Texas 75390-9039 +1 214 648 2827 Phone +1 214 648 6408 Fax christopher.gilpin-at-utsouthwestern.edu
Does anybody know a quick recipe for "grid glue" made out of scotch tape (or other sticky stuff) and some solvent for glueing films to TEM grids? Thanks in advance. -Karl
-- Karl Garsha Light Microscopy Specialist Imaging Technology Group Beckman Institute for Advanced Science and Technology University of Illinois at Urbana-Champaign 405 North Mathews Avenue Urbana, IL 61801 Office: B650J Phone: 217.244.6292 Fax: 217.244.6219 Mobile: 217.390.1874 www.itg.uiuc.edu
I am attempting to do a negative stain (which I know how to do) on a sample. the problem I'm having is that according to the protocol I have to glow discharge the grid before I use it.
I don't have access to a vacuum evaporator with a glow discharge unit. So what's a tech to do?
Any suggestions as to how to fake a glow are greatly appreciated.
Please help me to glimmer, glimmer. ;-)
Unable to glow,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
We have had a lot of success using ion beam sputter deposited Cr thin films for colloidal gold labeling of biological samples. Due to the nature of ion beam sputtered films we can precisely control the deposited thickness to less than 10 angstroms and still provide a uniform layer of Cr for charge reduction. The gold label on the sample provide high contrast in backscatter mode and the deposited Cr does not interfere with the signal.
If you would like additional information on this type of equipment or would like to see some examples I would be happy to provide this to you off-list. I hope this helps!
Best Regards,
Shane Roberts Applications Engineering Manager South Bay Technology, Inc. 1120 Via Callejon San Clemente, CA 92673 www.southbaytech.com roberts-at-southbaytech.com
DISCLAIMER: South Bay Technology produces equipment and supplies as described above and, therefore, has a vested interest in promoting their use.
-----Original Message----- } From: Chris Jeffree [mailto:c.jeffree-at-ed.ac.uk] Sent: Thursday, August 14, 2003 12:15 AM To: microscopy-at-sparc5.microscopy.com
James/Tom Yes, it is certainly my experience that gold palladium sputter coating quickly kills the BSE signal from 5 and 10nm colloidal gold. Carbon provides the best signal from small gold, but a thin sputtered chromium coating is also very good, and has the advantage of providing more fine topographical detail than can be obtained from carbon-coated specimens. I have used the Denton head in my Gatan Alto to do this, and have also had excellent results from specimens sputtered for us by Emitech.
Unfortunately, the techniques James suggests for reducing charging, while effective for reducing charging, may be just exactly what you want to avoid in BSE imaging of gold. Signal and signal/noise is often a problem, so one needs more beam current rather than less, and long record times, and it is therefore important that the coating and mounting technique is perfect. I often use the Analysis mode with the normal aperture 2/3 and and a relatively large spot size 5 on my Hitachi 4700, with the emission current increased to 15 or even 20µA, and find I need to be working at higher mags (say 20-100k) to get the best images.
Things to consider when optimizing carbon coating and specimen mounting are:
*Use rotary-tilt coating technique to get even coating all over. Static coating will leave a 3-d specimen with large uncoated shadow areas *Make sure the coating really is thick enough. A piece of white paper or masking tape placed near the specimens should be mid grey *Ensure the coating is in electrical continuity with the stub. If there is the slightest gap under the tissue block then paint it up with carbon dag. *If the specimens are large cubes (} 1mm) make sure the sides are adequately coated with carbon, and if in doubt dag them *Paint dag lines to the metal of the stub. Don't rely entirely on the conductivity of sticky carbon tabs, which is often not good enough.
Chris
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Facility
} ----- Original Message ----- } From: "firmiss" {firmiss-at-granicus.if.org} } To: "MSA ListServer" {Microscopy-at-sparc5.microscopy.com} } Sent: Wednesday, August 13, 2003 11:42 PM } Subject: Re: SEM - charging problems with bio samples } } } } -------------------------------------------------------------------- } ---- } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -------------------------------------------------------------------- } ---. } } } } } } On Sun, Aug 10, 2003 at 07:17:56PM -0500, Tom Phillips wrote: } } } } -------------------------------------------------------------------- } ---- } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -------------------------------------------------------------------- } ---. } } } } } } } } } We are relative novices on the SEM and are seeking advice on ways } to } } } minimize our charging problems. We would appreciate any tips from } the } } } mavens out there. } } } } } } We are looking at biological specimens (tissue explants) coated } either with } } } platinum or carbon (details below). The carbon coating protocol } is for } } } when we are trying to image at colloidal gold labeling on the } surface using } } } the BSE detector. The charging is much worse on the carbon coated } } } specimens compared to the platinum coated ones. I don't have a } feel for } } } how much of this problem is "normal" and one has to live with it } like so } } } much else in EM! But if anyone out there has some ideas or tricks } to } } } reduce charging, we are willing to try it. I would also be } interested in } } } anyone's nomination for a great reference book on SEM techniques. } Thanks. } } } Tom } } } } I only know a little bit about colloidal gold labeling but I would } think } } if you're using a BSE detector to look for the gold then I don't } think } } you'd want to coat the sample with gold or platinum (unless the gold } } particles are significant topographic features and you're using the } } BSE detector in 'topo' mode) -- that pretty much leaves you with } carbon } } coating -- more prone to charging but still usable. } } } } Here are some general SEM techniques you can try to reduce charging } for } } 'problem' samples. } } } } * Use smaller spot size } } * Use smaller aperature } } * Use a LOWER magnification if feasable } } * Increase scan rate } } } } All these basically reduce the rate at which electrons hit a given } area } } of your sample. } } } } If you're still having problems you may have to coat with more } carbon. } } } } I hope this helps. Good luck! } } } } -- } } James Firmiss ------ firmiss-at-granicus.if.org ------ } } "Let Sanity Prevail" } } }
We've been looking at the Nikon and Olympus TIRF systems.
Question 1: Is the 100X 1.65 N.A. objective really necessary?
Question 2: I would love to have an informal discussion off list with anybody who has experience with one or both of these systems. If I may speak with you, please contact me off list by email and we could write or set up a time that I could call you at your convenience.
Thanks! ____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
Light/ Confocal Microscopist: There is a full-time opening for a core facility light/confocal microscopist available immediately. The encumbent must demonstrate total facility with current technology and sample preparations. They will be responsible for 1) providing direct hands-on access to light microscopy in a core facility setting, 2) overseeing daily facility operations, 3) consulting with users on experiment design, materials and methods, and instrument optimization and assist in data acquisition and analysis, 4) troubleshooting problems with microscopes or samples and 5) assuring instrument integrity. Must maintain a current understanding of the microscopy field as necessary to implement new technologies on campus. Applicants must have a strong desire to participate actively as a member of a team, possess excellent written and oral communication skills, be comfortable working independently, communicate effectively with service users to understand their microscopy needs, be facile with PC and Mac computer platforms and demonstrate effective troubleshooting skills. M.S. in biological sciences with five years research experience in molecular biology, immunohistochemistry and cytology is required. Previous work in a core facility is preferred.
The Jackson Laboratory is one of the world's foremost centers for mammalian genetics research. Located in Bar Harbor, Maine, the lab is adjacent to Acadia National Park. Mountains, ocean, forests, lakes, and trails are all within walking distance. If you are looking for a more natural environment, this could be the opportunity you've been searching for.
Interested applicants should send cover letter and resume to: jax/210 Human Resources, Box 27 The Jackson Laboratory 600 Main Street Bar Harbor, Maine 04609 Fax: (207) 288-6106 Email (preferred option): jobs-at-jax.org The Jackson Laboratory is an Affirmative Action/Equal Opportunity Employer.
Lesley S. Bechtold Supervisor, Biological Imaging The Jackson Laboratory 600 Main St. Bar Harbor, ME 04609 207-288-6191
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Paula Sicurello wrote: ================================================================== I am attempting to do a negative stain (which I know how to do) on a sample. the problem I'm having is that according to the protocol I have to glow discharge the grid before I use it.
I don't have access to a vacuum evaporator with a glow discharge unit. So what's a tech to do?
Any suggestions as to how to fake a glow are greatly appreciated.
Please help me to glimmer, glimmer. ;-)
Unable to glow, =================================================================== If this is the kind of treatment one would apply to carbon coated grids to make them more hydrophilic, we can produce the same effect in the SPI Plasma Prep II Plasma Etcher but with an "air" plasma instead of an oxygen plasma (using oxygen would change your sample, actually it would probably etch away and effectively destroy your sample). The Plasma Prep II unit is shown on URL http://www.2spi.com/catalog/instruments/etchers1.shtml
We would be happy to do a "demo" of the technique for you if you sent us your grids in a grid box. Contact me off-line and I will give you shipping instructions. When this technique is used to make carbon coated grids hydrophilic, the effect seems to last something like 90 days.
Disclaimer: SPI Supplies is the manufacturer of the Plasma Prep II plasma etcher/cleaner/asher so we would have a vested interest in seeing more of them sold.....
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id PAA16630 for dist-Microscopy; Thu, 14 Aug 2003 15:24:37 -0500 (CDT) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id PAA16626 for "MicroscopyFilteredEmail4-at-msa.microscopy.com"; Thu, 14 Aug 2003 15:24:06 -0500 (CDT) Received: from web40206.mail.yahoo.com (web40206.mail.yahoo.com [66.218.78.67]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id PAA16618 for {Microscopy-at-sparc5.microscopy.com} ; Thu, 14 Aug 2003 15:23:53 -0500 (CDT) Message-ID: {20030814202245.40055.qmail-at-web40206.mail.yahoo.com} Received: from [137.131.240.13] by web40206.mail.yahoo.com via HTTP; Thu, 14 Aug 2003 13:22:45 PDT
Dear Steve and other microscopist,
The Formvar on your Carbon film is hard to be cleaned completely. The rest of Formvar could cause several problems, such as charging, poor stigmatism, drifting of specimen and even chemical reaction with your sample in EM.
I suggest you to try the pure carbon film coated grid, therefore you don't need to clean the Formvar on your grids. This carbon film didn't touch with any plastics while it was made. The pure carbon film is floated on the surface of water and sit on the cleaned grids.
We do produce this kind of grid. For more information, please check our website at
http://www.grid-tech.com/
Good luck.
Wendy ===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
--- Steve Parry {sparry-at-cmm.uwa.edu.au} wrote: } } } I have recently switched to Formvar in Diethylene } Chloride as a base } for making carbon coated grids, because I was told } that I would get } bettter continuous films, the problem I am having is } that I cannot } get rid of the forvar after coating. } Does anyone out there have a method for making C } coated grids that } works everytime?? I am getting to the end of my } patience and have } wasted an awful lot of grids over the last few } months. } } Thanks } } Steve Parry } -- } Steve Parry } Centre for Microscopy & Microanalysis, } (M010) } The University of Western Australia, } 35 Sterling Highway, Crawley, } WA 6009, Australia. } } Fax: +61-8-9380-1087 } Email: sparry-at-cmm.uwa.edu.au } Phone: +61-8-9380-8057 [24 hour voicemail } attached] } }
===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
Karl Garsha {garsha-at-itg.uiuc.edu} ASKED THE FOLLOWING QUESTION:
} Does anybody know a quick recipe for "grid glue" made out of scotch } tape (or other sticky stuff) and some solvent for glueing films to } TEM grids? Thanks in advance. } -Karl } RESPONSE:
GRID GLUE can be prepared by dissolving the adhesive from 2 inches of Scotch Brand transparent tape in 10 ml of ethylene dichloride. Please be sure to use the transparent tape (old style) rather than the Magik Tape since the adhesives are apparently different. I believe that chloroform can also be used as a solvent.
It might also be posible (and I invite microscopy vendors to chime in) to use the commercially prepared Grid-Stick Glue (EMS sells it, for example) but it will have to be diluted.... I have not tried this, however.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
We are getting rid of a functioning Philips 505 SEM. Anyone with a 500 series scope that might like parts let me know ASAP. SEI,BEI,CL,W-hairpin (boxes of filaments), LAB6 (sorry no filaments - got IG pumps), pneumatic valves, bit and pieces, etc. etc.. . .
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
There are several possibilities here, but none of them are as good as the real thing.
1. Put your grids in the chamber of a sputter coater but cover the grids with a "tent" of filter paper. The tent should completely cover over the grids so that few of the metal particles will strike it but the argon ions will. Alternatively, you could turn the slide containing the grids upside down so that they face away from the target. Just be sure to leave some space underneath, so the ions can contact the grids. Activate the sputtering process so that you see the plasma for 40-50 sec. You might have to experiment here so that you minimize the presence of metal. If you have a carbon sputterer, I believe that would be the best.
2. You might try a dozen hits of the grids with one of the electrostatic guns (used by photographers). This sometimes does work.
3. Finally, and don't ask why this works, just leaving the grids for several days inside of a refrigerator will cause them to become hydrophilic. I am guessing that the trapped organics (odors, fumes, etc.) are depositing onto the active surface of the carbon. This is what happens when you place sodium bicarb in the refrig to trap odors.
Let us know which work best for you.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
A couple of times I just dissolved cellotape in chloroform - as much as could be squeezed into a small 10 ml sample bottle. Saturated solution? Not scientific but it helped when needed!
Keith Ryan Marine Biological Association of the UK & University of Plymouth, UK
----- Original Message ----- } From: "Karl Garsha" {garsha-at-itg.uiuc.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, August 14, 2003 4:19 PM
Hello, Just another question about our Electroscan E3 ESEM. Lately, we've noticed that when you start up the main console power, the right screen remains grey and doesn't load the microscope software such as the vacuum/gun/image menus. The left screen has a scale bar on it, but nothing works. I have to power down and then power up, sometimes 3x for it to boot up correctly.
Anyone have any advice where I should look for problems? Thanks for the help.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I make a general purpose glue by stuffing several feet of Scotch Double Stick tape (other types would probably work as well) into a 100ml bottle then adding about 50 ml of heptane. Shake the bottle several times a day and in a few days you'll have a nice solution that will leave a sticky residue when it dries. This adhesive may not be the best to use in an electron microscope since it may outgas and contaminate your system--do some tests. Back in the 1970's there was a paper given at the MSA (EMSA) meeting describing a solution of Neoprene W in chloroform. It worked well with minimal outgassing but the only source of the Neoprene was DuPont. I have a few pieces buried somewhere but hopefully someone else has a beter source.
On Thu, 14 Aug 2003 10:19:19 -0500 Karl Garsha {garsha-at-itg.uiuc.edu} wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy } Society of America
Larry Ackerman Advanced Imaging Lab/Dr. John Sedat/Dr. David Agard Dept. of Biochemistry & Biophysics University of California San Francisco 600-16th Street, Box 2140 San Francisco, CA 94143
This might be due to a bad connection / malfunctioning of a SCSI-card inserted in your computer to connect the computer with the microscope. If one connection fails, ik can interfere other connections too. Try to see how the pc starts up when you disconnect the microscope from it. Then start up again after connecting one by one.
Another possibility is, if you have SCSI-hardware, there are conflicts with the startup-sequence. In other words, SCSI-hardware should start up in a particular order (one has to start up BEFORE/AFTER another). If you just added a new piece of hardware, the numbering might be conflicting.
Sven Terclavers
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We sometimes have the same problem with our E-3 scope. In our case it occurs because the disk drive does not begin to read the disk. In that case the yellow light on the disk drive will not light up when you start the microscope.
We have developed a special trick for our E-3 scope, where we give the disk a slight pressure to the left at the end of the insertion into the drive. When the disk has been inserted that way, it can usually be read by the drive.
-----Original Message----- } From: Gordon Vrololjak [mailto:gvrdolja-at-nature.Berkeley.EDU] Sent: 14. august 2003 23:56 To: Microscopy-at-sparc5.microscopy.com
Hello, Just another question about our Electroscan E3 ESEM. Lately, we've noticed that when you start up the main console power, the right screen remains grey and doesn't load the microscope software such as the vacuum/gun/image menus. The left screen has a scale bar on it, but nothing works. I have to power down and then power up, sometimes 3x for it to boot up correctly.
Anyone have any advice where I should look for problems? Thanks for the help.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
I use double sided tape in chloroform (a ball of tape that is just covered by the chloroform). Shake the bottle for 20 sec. and remove what is left of the tape (you want the glue off of the tape, not the plastic). Not too scientific, but it works. David
On Thursday, August 14, 2003, at 11:19 AM, Karl Garsha wrote:
} ----------------------------------------------------------------------- } - } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } ----------------------------------------------------------------------- } . } } } Does anybody know a quick recipe for "grid glue" made out of scotch } tape (or other sticky stuff) and some solvent for glueing films to TEM } grids? Thanks in advance. } -Karl } } -- } Karl Garsha } Light Microscopy Specialist } Imaging Technology Group } Beckman Institute for Advanced Science and Technology } University of Illinois at Urbana-Champaign } 405 North Mathews Avenue } Urbana, IL 61801 } Office: B650J } Phone: 217.244.6292 } Fax: 217.244.6219 } Mobile: 217.390.1874 } www.itg.uiuc.edu } } } } ____________________
David Elliott Ph.D.
Yale University School of Medicine Campus Address: TAC S160 333 Cedar Street PO Box 208022 New Haven, CT 06520-8022
Re method 3. placing grids in a fridge, do you put them in on filter paper in a petri dish with the lid on, for instance?
Dave
On Thu, 14 Aug 2003 15:45:26 -0500 "John J. Bozzola" {bozzola-at-siu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Paula, } } There are several possibilities here, but none of them are as good as } the real thing. } } 1. Put your grids in the chamber of a sputter coater but cover the } grids with a "tent" of filter paper. The tent should completely cover } over the grids so that few of the metal particles will strike it but } the argon ions will. Alternatively, you could turn the slide } containing the grids upside down so that they face away from the } target. Just be sure to leave some space underneath, so the ions can } contact the grids. Activate the sputtering process so that you see } the plasma for 40-50 sec. You might have to experiment here so that } you minimize the presence of metal. If you have a carbon sputterer, I } believe that would be the best. } } 2. You might try a dozen hits of the grids with one of the } electrostatic guns (used by photographers). This sometimes does work. } } 3. Finally, and don't ask why this works, just leaving the grids for } several days inside of a refrigerator will cause them to become } hydrophilic. I am guessing that the trapped organics (odors, fumes, } etc.) are depositing onto the active surface of the carbon. This is } what happens when you place sodium bicarb in the refrig to trap odors. } } Let us know which work best for you. } } JB } } ############################################################## } John J. Bozzola, Ph.D., Director } I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) } 750 Communications Drive - MC 4402 } Southern Illinois University } Carbondale, IL 62901 U.S.A. } Phone: 618-453-3730 } Email: bozzola-at-siu.edu } Web: http://www.siu.edu/~image/ } ############################################################## } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
We have used "grid pens" with glue with great success for our immunocytochemistry work. Initially, we had put up with LR White and Unicryl sections looking like a ragged piece of kleenex in the TEM. Then we moved to formvar/carbon grids, but had severe problems with "stickiness" causing high background labeling, apparently due to both antibody binding and mechanical trapping of the gold conjugate, not to mention various other folding and layering artifacts. Finally, after a suggestion from someone on this list, we tried the pens. Now our sections remain largely intact, the background problem is under control, and we are much happier campers. We do not dilute the glue in the pens, but just put one dab on grids on filter paper and let them dry a couple minutes before picking up our sections.
Cheers, Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: John J. Bozzola [mailto:bozzola-at-siu.edu] Sent: Thursday, August 14, 2003 3:29 PM To: Microscopy-at-sparc5.microscopy.com
Karl Garsha {garsha-at-itg.uiuc.edu} ASKED THE FOLLOWING QUESTION:
} Does anybody know a quick recipe for "grid glue" made out of scotch } tape (or other sticky stuff) and some solvent for glueing films to } TEM grids? Thanks in advance. } -Karl } RESPONSE:
GRID GLUE can be prepared by dissolving the adhesive from 2 inches of Scotch Brand transparent tape in 10 ml of ethylene dichloride. Please be sure to use the transparent tape (old style) rather than the Magik Tape since the adhesives are apparently different. I believe that chloroform can also be used as a solvent.
It might also be posible (and I invite microscopy vendors to chime in) to use the commercially prepared Grid-Stick Glue (EMS sells it, for example) but it will have to be diluted.... I have not tried this, however.
JB
############################################################## John J. Bozzola, Ph.D., Director I.M.A.G.E. (Integrated Microscopy & Graphics Expertise) 750 Communications Drive - MC 4402 Southern Illinois University Carbondale, IL 62901 U.S.A. Phone: 618-453-3730 Email: bozzola-at-siu.edu ##############################################################
first, have you tried a standard negative stain preparation? a lot of the time there is enough protein present in the sample to overcome the hydrophobicity. just leave the sample on the grid for about a minute.
second, try an airfuge, EM-90 preparation. the combination of centrifugal force for 1/2 hr plus protein present in the preparation usually overcomes the need for glow discharge.
third, try pre-treating the grids with either poly-L-lysine or Alcian blue. use a 1% solution, float the grids on the solution for 1-2 minutes, wick off the excess, then let dry
one of these should work
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
Michael, The 1.65 NA objective is not absolutely necessary, but as you aware TIRF needs higher NA (NA} n). But you can obtain fairly good images with 1.45 NA lenses available now with Nikon and Zeiss. The 1.65 NA available only with Olympus as of now and you may need to pay a higher price for that. I had an introductory practical course experience only with Olympus system with both 1.45 and 1.65 lenses. If you need any other information please let me know by email.
Shiv
Mayandi Sivaguru, Ph.D., Associate Director Molecular Cytology Core Facility Molecular Biology Program 2, Tucker Hall University of Missouri Columbia, MO 65211-7400 sivagurum-at-missouri.edu
Voice: 1-573-882-4895 Fax: 1-573-882-0123
www.biotech.missouri.edu/mcc/
At 12:42 PM 8/14/03 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It came to my mind, that exposure of the grids to the strong UV (real UV) light do the trick also. Effect will vary depends from the UV source and intensity. As I remember, grids are good for about 40 min after all. Personally, I prefer to use poly-lysine treatment. In my hands it works nearly the same as glow discharge. Even better, because it does not made surface rough. Sergey
At 01:45 PM 8/14/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (sswaffe-at-abv.bg) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Wednesday, August 13, 2003 at 02:56:55 ---------------------------------------------------------------------------
Email: sswaffe-at-abv.bg Name: Veselin Andreev
Organization: 54 "st.Ivan Rilski"
Education: 6-8th Grade Middle School
Location: Sofia Bulgaria
Question: How can I prepare permanent slides for examination under microscope?What materials should I use for preserving the speciment for a very long time?
Veselin- There are commercial preparations you can buy, but you can also use clear nail polish if you do not have access to these cements. But first- you must remove the water from your specimen, by soaking in alcohol. First water plus alcohol, then gradually reducing the water content. Making permanent slides takes a different process depending upon what the specimen is. What are you mounting on the slide? Rgds, Mike Shaw } } Email: sswaffe-at-abv.bg } Name: Veselin Andreev } } Organization: 54 "st.Ivan Rilski" } } Education: 6-8th Grade Middle School } } Location: Sofia Bulgaria } } Question: How can I prepare permanent slides for examination under microscope?What materials should I use for preserving the speciment for a very long time? } } ------------------------------------------------------------ } ---------------
Richard I use Electron Microscopy, 2nd edition by John Bozzola and Lonnie Russell for my graduate TEM course. It is an excellent book and my students really enjoy it, especially the survey of biological ultrastructure at the back.
My favorite is "A beginners handbook in Biological Transmission Electron Microscopy", by Brenda S. Weakley, Chruchill Livingstone is the publisher. My second edition is 1981, there may be a more recent edition. "Biological Techinques in Electron Microscopy" by Clinton Dawes is also good but my copy is so old (1971) it may be out of print.
Geoff
Richard Gardiner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
You can easily use the method described by Mike to make about 50 pure carbon grids at a time (much cheaper than buying if you already have the equipment).
You place about 50 grids on a square piece of filter paper lying on a little "table" made of wire mesh under the water surface. To get the grids under water make them wet first and push them through the surface vertically. Then you lower the water surface by removing water from underneath (for example with a syringe or a plastic tube with a valve) to place the carbon film (floated off the mica) on top of the grids. (Of course the carbon film has to be in one piece in this case.) Push the carbon into position with something blunt (a paddle ? :)
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em _______________________________________
} } Steve, } } When I want just carbon as a substrate I will } } coat freshly cleaved mica with carbon in a metal } } evaporator. This allows me to dictate the carbon } } thickness and I have no trouble floating off grid } } size squares(prescored)in a drop of water. Picking up the } } carbon with glow discharged grids (flat side) } } is a snap. Make sure everything is clean. } } This should give you thin clean substrates. } } Mike D.
I've used the Bozzola and Russell textbook entitled "Electron Microscopy: Principles and Techniques for Electron Microscopy" since the first edition was published in 1992. A second edition was published in 1999. It was especially suited to my needs as I taught both an SEM and a TEM course each year - primarily to undergraduate students. The students were able to use the same text for both courses. The authors provide a good general background on theory, and an excellent treatment of procedures and techniques for both types of microscopy. Although there may be other texts out there that are suitable as well, I highly recommend this one as a text to teach from.
Doug Bray
Richard Gardiner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } Hello: } } I was wondering what text books people are using to teach either } Undergraduate or Graduate courses in Biological TEM. } } Richard } Gardiner
Bozzola, John J., Russell, Lonie D. - Electron Microscopy - Principles and Techniques for Biologists.
Alice Dohnalkova Environmental Microbiology Pacific Northwest National Laboratory MS P7-50 Richland, WA 99352 tel. (509) 372-0692 office (509) 376-3654 TEM lab fax (509) 376-1321
} -----Original Message----- } From: Richard Gardiner [SMTP:rbgardiner-at-rogers.com] } Sent: Saturday, August 16, 2003 12:09 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: EM Textbooks Used } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello: } } I was wondering what text books people are using to teach either } Undergraduate or Graduate courses in Biological TEM. } } Richard } Gardiner } }
Hi, We use PTA to stain thin sections of human skin for collagen. I make up a fresh solution of 1% PTA in dH20. Mix thoroughly and filter twice through a #1 Whatman. Leave the pH acid. Don't adjust it toward neutral (it won't stain as well). Stain sections 30 mimutes. staining sequence should be: 1) PTA 2) UA 3) PB
Robert Underwood Dermatology Research Center University of Washington
On Mon, 18 Aug 2003, Dusevich, Vladimir wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need a protocol for staining collagen thin sections with } phosphotungstic acid. } } Thanks a lot, } } Vladimir } } Vladimir M. Dusevich, Ph.D. } Electron Microscope Lab Manager } 3127 School of Dentistry } 650 E. 25th Street } Kansas City, MO 64108-2784 } } Phone: (816) 235-2072 } Fax: (816) 235-5524 } Web: http://www.umkc.edu/dentistry/microscopy } } } }
With many of you looking for extra money for microscopy labs and the recent convergence of microscopy and spectroscopy, I thought this small grant program might be of interest.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^ Need class-room sized quantities of Optimizing Light Microscopy? Call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^
ANNOUNCEMENT
2004 Pittsburgh Conference Memorial National College Grant Program Eligibility Criteria
The Pittsburgh Conference on Analytical Chemistry and Applied Spectroscopy (a Pennsylvania non-profit Corporation) and its co-sponsoring technical societies, The Society for Analytical Chemists of Pittsburgh (SACP) and The Spectroscopy Society of Pittsburgh (SSP) proudly announce the 2004 Pittsburgh Conference Memorial National College Grants (PCMNCG) Program.
Grants will be awarded to small college science departments for the purchase of scientific equipment, audio-visual or other teaching aids, and/or library materials for use in the teaching of science at the undergraduate level.
Based on submitted proposals, at least twelve (12) colleges will be selected to receive grants, each grant having a maximum of $9,000.
To be eligible for an award, schools must meet the following criteria. 1. Enrollment must not exceed 5000 full-time students.
2. No more than 25% of the operating budget may come from national or state governments. NOTES Two-year community colleges sponsored by political subdivisions of a state are not bound by criteria 1 and 2.
Schools should not expect funding from other programs sponsored by SACP and SSP (if they receive a PCMNCG grant). 3. Requests that support undergraduate research are allowed. However, requests for equipment to be used solely for non-instructional research purposes shall not be funded.
4. Awards may be used as part of Matching Grant programs; use of matching funds to increase the grants impact is encouraged.
5. Previous awardee colleges are ineligible for the PCMNCG program for a three-year period following receipt of the grant (awardee colleges from 2001, 2002 and 2003 are not eligible for the 2004 program).
6. Grant applications and proposals must be received no later than December 1, 2003. Faculty members may obtain application forms and additional information from:
Rita M. Windisch, Ph.D. The Pittsburgh Conference PCMNCG 300 Penn Center Blvd., Suite 332 Pittsburgh, PA 15235-5503 Telephone (412) 825-3220, Ext. 204 FAX (412) 825-3224 E-mail: windisch-at-pittcon.org
or from the Pittsburgh Conference Website: www.pittcon.org Announcement of the awards will be made by February 2004. Schools chosen will join the list of over 200 institutions honored since the inception of this program in 1974.
I think Dawes is still available from Ladd Research. But I prefer Bozzola and Russell for a text, and Maunsbach and Afzelius "Biomedical Electron Micropscopy" for lots of EMs comparing fixatives, buffers, embedding media, and so forth.
Phil
} My favorite is "A beginners handbook in Biological Transmission } Electron Microscopy", by Brenda S. Weakley, Chruchill Livingstone } is the publisher. My second edition is 1981, there may be a more } recent edition. "Biological Techinques in Electron Microscopy" by } Clinton Dawes is also good but my copy is so old (1971) it may be } out of print. } } Geoff } } Richard Gardiner wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Phillip Your way to mount carbon on the grids has one very serious disadvantage: when you mount carbon this way, you will use "air-side" of carbon as a working surface. This side is usually more grainy and dirty. The whole idea of using carbon from mica is to use "mica-side" of carbon as a working surface. The idea here is that the carbon face toward mica is more uniform and perfectly clean (it was not exposured to the air). In order to use "good" side of carbon, you have to put grids on floated carbon and then pick the grids with Parafilm for instance. Similarly, you need to do the same for plastic film created on water - working side is "water-side", not air. Only one exception I do know is Formvar film created on the glass. Glass is not such clean and perfect as mica or water surface, therefore, you need to use "air-side". Then your technique will work perfectly. I also use your technique to mount holey film on the grids. Best wishes, Sergey.
At 04:07 PM 8/18/03 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ssamuelsson-at-eyetk.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 18, 2003 at 14:09:26 ---------------------------------------------------------------------------
Email: ssamuelsson-at-eyetk.com Name: Steve Samuelsson
Organization: Eyetech Pharmaceuticals
Title-Subject: Whole Mounts
Question: Regarding growing endothelial cells on nickel grids for whole mount TEM analysis: our luck is inconsistent and we now need to dig further for the reason. 200 and 300 mesh nickel grids are coated with ~0.5% formvar in ethylene dichloride (we also tried chloroform) and picked up on a 13mm cover slip. These are allowed to dry and age a day or longer. Prior to plating, the slips are UV sterilized 10-15', incubated in 1% gelatin ~45', washed 1x with PBS, culture medium added followed by cells. Our issue is incomplete (or downright lousy) growth of cells on the grids. Cells are robust and growing fine on the bottom of the 24 well plate so we suspect something with the grids. My first thought was that we needed carbon coating of the grids but have some trouble with this explanation. Second thought was that the solvent was still coming out of the films and poisoning the cells; thus the 'aging' issue with the cover slips. Any suggestions?
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (hafeez189-at-yahoo.com) from http://www.msa.microscopy.org/Ask-A-Microscopist.html on Monday, August 18, 2003 at 14:38:56 ---------------------------------------------------------------------------
Email: hafeez189-at-yahoo.com Name: Hafeez Anwar
Organization: University of Agriculture, Faisalabad, Pakistan
Education: Graduate College
Location: Pakistan
Question: hi, i am working on optical polarization with hot stage and i am studying the morphology of polymer blends. my question is that why the colors of the sample change when we rotate the analyzer ?(kindly ,explain in detail)
A user has the following question. Please reply off line to her at: DCOLLISO-at-glcc.com (Donna Collison)
Does anyone have experience in staining 'toughened nylon' for backscattered compositional analysis. Toughened nylon as we are describing it is nylon having butadiene rubber phases within the nylon matrix. We need to determine were an antimony component is within the formulation. So far we have placed fractured pieces of this material directly in osmium tetroxide for a 24 hour period and was not able to detect any contrast. Any suggestions and help would be greatly appreciated.
Many thanks, Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
I need a little advice on safely examining plants infected with Salmonella sp. I have a user who wants to use cryofixation of melon and tomato plants that have been inoculated with the Salmonella sp. bacterium. She would like to bring the inoculated plants into the EM facility, dissect a small bit of the plant surface and then do a conventional cryo prep and examine in the SEM. I am not familiar with the safest method of examining pathogens in the SEM. Can the bacteria survive cryo fixation? What is the best method of cleaning the scope after use? What about the instruments used for dissection? As you can see, I have a lot of questions and before we proceed with this project I need to satisfy myself that we are doing this correctly.
Thanks for you help.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://microscopy.mcb.ucdavis.edu raharris-at-ucdavis.edu
On Mon, 18 Aug 2003 17:59:25 -0500 by way of Ask-A-Microscopist {ssamuelsson-at-eyetk.com} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ssamuelsson-at-eyetk.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 18, 2003 at 14:09:26 } --------------------------------------------------------------------------- } } Email: ssamuelsson-at-eyetk.com } Name: Steve Samuelsson } } Organization: Eyetech Pharmaceuticals } } Title-Subject: Whole Mounts } } Question: Regarding growing endothelial cells on nickel grids for whole mount TEM analysis: our luck is inconsistent and we now need to dig further for the reason. 200 and 300 mesh nickel grids are coated with ~0.5% formvar in ethylene dichloride (we also tried chloroform) and picked up on a 13mm cover slip. These are allowed to dry and age a day or longer. Prior to plating, the slips are UV sterilized 10-15', incubated in 1% gelatin ~45', washed 1x with PBS, culture medium added followed by cells. Our issue is incomplete (or downright lousy) growth of cells on the grids. Cells are robust and growing fine on the bottom of the 24 well plate so we suspect something with the grids. My first thought was that we needed carbon coating of the grids but have some trouble with this explanation. Second thought was that the solvent was still coming out of the films and poisoning the cells; thus the 'aging' issue with the cover slips. Any suggestions? } } } --------------------------------------------------------------------------- } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
My own experience has been that only gold or titanium grids give consistent results with this procedure. Both copper and nickel tend to be quite toxic.
Roger Moretz, Ph.D. Dept of Toxicology BI Pharmaceuticals Ridgefield, CT
-- Where the world is only slightly less weird than it actually is. } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Could the nickel be toxic? Why not try gold? } } Dave } } } On Mon, 18 Aug 2003 17:59:25 -0500 by way of } Ask-A-Microscopist {ssamuelsson-at-eyetk.com} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Below is the result of your feedback form (NJZFM-ultra-55). It was submitted } by (ssamuelsson-at-eyetk.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, } August 18, 2003 at 14:09:26 } } --------------------------------------------------------------------------- } } } } Email: ssamuelsson-at-eyetk.com } } Name: Steve Samuelsson } } } } Organization: Eyetech Pharmaceuticals } } } } Title-Subject: Whole Mounts } } } } Question: Regarding growing endothelial cells on nickel grids for whole mount } TEM analysis: our luck is inconsistent and we now need to dig further for the } reason. 200 and 300 mesh nickel grids are coated with ~0.5% formvar in ethylene } dichloride (we also tried chloroform) and picked up on a 13mm cover slip. These } are allowed to dry and age a day or longer. Prior to plating, the slips are UV } sterilized 10-15', incubated in 1% gelatin ~45', washed 1x with PBS, culture } medium added followed by cells. Our issue is incomplete (or downright lousy) } growth of cells on the grids. Cells are robust and growing fine on the bottom } of the 24 well plate so we suspect something with the grids. My first thought } was that we needed carbon coating of the grids but have some trouble with this } explanation. Second thought was that the solvent was still coming out of the } films and poisoning the cells; thus the 'aging' issue with the cover slips. Any } suggestions? } } } } } } --------------------------------------------------------------------------- } } } } } } } } This incoming email to UWE has been independently scanned for viruses and any } virus detected has been removed using McAfee anti-virus software } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } }
Suppliers are, in my experience, reluctant to comment on the problem potential hazards of non chemically fixed microorganisms in cryoSEM orlow vacuum/ESEM.
I have seen a paper showing that some bacteria (which species?) survive cryoSEM sessions. In theory they could get pumped into your rotary pump and out of the room (comments anyone?).
A colleague looked for presence of bacteria in the chamber after ESEM and found little, I think. (I will e-mail her for comment).
Here if a colleague can show the bacteria are harmless we would consider looking at them unfixed. If they are pathogenic I insist they are fixed first.
Your colleague should advise on cleaning tools. One of our users cleaned forceps in 90% ethanol. You can wipe over the stage and chamber but what about the column?
I hope you have started an interesting debate on a topic we mull over here periodically.
Dave
On Mon, 18 Aug 2003 16:52:52 -0700 "Rick A. Harris" {raharris-at-ucdavis.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I need a little advice on safely examining plants infected with Salmonella } sp. I have a user who wants to use cryofixation of melon and tomato plants } that have been inoculated with the Salmonella sp. bacterium. She would like } to bring the inoculated plants into the EM facility, dissect a small bit of } the plant surface and then do a conventional cryo prep and examine in the } SEM. I am not familiar with the safest method of examining pathogens in the } SEM. Can the bacteria survive cryo fixation? What is the best method of } cleaning the scope after use? What about the instruments used for } dissection? As you can see, I have a lot of questions and before we proceed } with this project I need to satisfy myself that we are doing this correctly. } } Thanks for you help. } } Rick A. Harris, Director } Microscopy and Imaging Facility } Section of Molecular and Cellular Biology } 1241 Life Sciences Addition } University of California } Davis, CA } 530 752 2914 } http://microscopy.mcb.ucdavis.edu } raharris-at-ucdavis.edu } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I highly recommend Bozzola & Russell's Electron Microscopy 2nd ed. I use it for my graduate level EM course.
Cheers,
Soumitra
************************************************************* Soumitra Ghoshroy College Associate Professor, Biology Director, Electron Microscopy Lab Box 3EML New Mexico State University Las Cruces, NM 88003 Tel: 505-646-3268 (office), 646-3283 (lab) Fax: 505-646-3282 e-mail:sghoshro-at-nmsu.edu http://www.cs.nmsu.edu/~biology/Faculty%20&%20Staff/Ghoshroy/Ghoshroy.htm http://confocal.nmsu.edu/eml
On Sat, 16 Aug 2003, Richard Gardiner wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hello: } } I was wondering what text books people are using to teach either } Undergraduate or Graduate courses in Biological TEM. } } Richard } Gardiner } } }
I have a lot of trouble making too Epon/Araldite in this weather of high humidity. Although our hospital is air conditioned, it still seems to leave a lot of humidity in the air [read: it's almost useless], and it seems that we always having a lot of trouble infiltrating and getting the thin sections to stay together in this sort of weather. Our thin sections break either in the boat when we are cutting, or rapidly under the beam of the microscope.
Is there any way of modifying the formula of our plastic to compensate for this problem?
This is the formula that we presently use to make our plastic:
On Monday, August 18, 2003, at 04:52 PM, Rick A. Harris wrote:
} I need a little advice on safely examining plants infected with } Salmonella } sp. I have a user who wants to use cryofixation of melon and tomato } plants } that have been inoculated with the Salmonella sp. bacterium. She } would like } to bring the inoculated plants into the EM facility, dissect a small } bit of } the plant surface and then do a conventional cryo prep and examine in } the } SEM. I am not familiar with the safest method of examining pathogens } in the } SEM. Can the bacteria survive cryo fixation? What is the best method } of } cleaning the scope after use? What about the instruments used for } dissection? As you can see, I have a lot of questions and before we } proceed } with this project I need to satisfy myself that we are doing this } correctly. } Dear Rick, We do some cryo-TEM on (mild) pathogens, and our safety-office-approved protocol is to do the preparation--growth and freezing--in a biosafety hood. We are allowed to transfer the grids, which are kept under LN2, to the cryostage as with any other specimen, then the grids are dropped into bleach after examination. You should assume that Salmonella will survive cryofixation, and treat your specimens, tools, stubs, etc. as potential sources of infection. Check with your safety office for the best sterilization method for Salmonella, but bleach or detergent will usually disrupt bacteria. I'm not at all certain what would be best for the inside of the SEM, but, if your specimens remain frozen for the duration of their time inside the scope, you may be able to avoid putting bleach or detergent into your instrument. Good luck. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
On Monday, August 18, 2003, at 04:00 PM, by way of Ask-A-Microscopist wrote:
} Question: hi, i am working on optical polarization with hot stage and } i am studying the morphology of polymer blends. } my question is that why the colors of the sample change when we rotate } the analyzer ?(kindly ,explain in detail) } Dear Hafeez, Your sample causes a rotation of the polarization vector of the light passing through it. The amount of the rotation is proportional to the thickness of the sample, and it will vary with the wavelength of the light. The phenomenon is called "optical rotary dispersion" (if I remember my optics correctly), and the details should be available in a physics text. Consider a simple case. If you are looking through a slab of material with uniform thickness, the light entering the sample will be polarized, say, at 12 o'clock. As it passes through the slab, the polarization of each wavelength is rotated to a different extent, say, red to 5 o'clock, green to 6 o'clock, and blue to 7 o'clock. When the analyzer is oriented at 5 o'clock, the sample will appear red, etc. If the sample thickness varies, the apparent color at a given analyzer position will vary with thickness, and, as the analyzer orientation changes, the colors will shift. If your specimen is wedge-shaped, you should see the colors in the same order as in a rainbow--assuming that the dispersion is monotonic with wavelength, as it usually is--and the colors will move pretty much in unison along the slope of the wedge as the analyzer is rotated. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
Epon is somewhat miscible with water, a colleague regularly goes from 95% to an Epon:alcohol mix, so humidity may not be the problem. First I would try all fresh plastics, Epon, Araldite and especially DMP-30 which has a short shelf life. You could also try longer times (or smaller pieces of tissue) and agitation to be sure infiltration is not the problem. It could be that your plastics have gotten 'tired' just as the weather has become more humid?
Geoff
Garry Burgess wrote:
} I have a lot of trouble making too Epon/Araldite in this weather of high } humidity. Although our hospital is air conditioned, it still seems to leave } a lot of humidity in the air [read: it's almost useless], and it seems that } we always having a lot of trouble infiltrating and getting the thin sections } to stay together in this sort of weather. Our thin sections break either in } the boat when we are cutting, or rapidly under the beam of the microscope. } } Is there any way of modifying the formula of our plastic to compensate for } this problem? } } This is the formula that we presently use to make our plastic: } } Araldite: 65 grams } Epon [JEMBED 812 Epon relacement]: 81 grams } DDSA: 232 grams } DMP-30: 7.2 grams. } } } Garry Burgess } Charge Technologist } Department of Pathology - EM Laboratory } Health Sciences Centre } Winnipeg } } }
-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
One thing I've found that at least helps is to pre-cook the embedding molds to sort of dry them out. Also, store the resin components under vacuum. Don't know about changing your recipe....
Good luck!
Tamara
On Tue, 19 Aug 2003, Garry Burgess wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I have a lot of trouble making too Epon/Araldite in this weather of high } humidity. Although our hospital is air conditioned, it still seems to leave } a lot of humidity in the air [read: it's almost useless], and it seems that } we always having a lot of trouble infiltrating and getting the thin sections } to stay together in this sort of weather. Our thin sections break either in } the boat when we are cutting, or rapidly under the beam of the microscope. } } Is there any way of modifying the formula of our plastic to compensate for } this problem? } } This is the formula that we presently use to make our plastic: } } Araldite: 65 grams } Epon [JEMBED 812 Epon relacement]: 81 grams } DDSA: 232 grams } DMP-30: 7.2 grams. } } } Garry Burgess } Charge Technologist } Department of Pathology - EM Laboratory } Health Sciences Centre } Winnipeg } }
|--------------------------------------------------| Tamara Howard Department of Cell Biology and Physiology University of New Mexico - Health Sciences Center Albuquerque, NM 87131 thoward-at-unm.edu |--------------------------------------------------|
Now that the safety nazis have forced us to wear face shields, smocks and gloves for handling liquid nitrogen, they want us to wear safety glasses while doing wedge polishing, which entails frequent looking through a microscope to judge the progress. The concern is that some polish slurry 'could' splash into the eyes of a technician. Of course the safety glasses really get in the way when looking through a microscope, and our technicians are really not happy about this. I would like to hear as many opinions as possible on whether there is significant likelyhood of such a splash, and if polishing slurry, like Siton or Glanzox really poses a hazard to the eyes of a person using it. Thanks.
This is a simple matter of your company doing risk assessment and covering their butts to prevent law suits and higher insurance premiums. I don't think it matters if it is silicon polish, rock chips off a jack hammer or any other flying material, a company/institution wants to lessen liability and increase safety. By telling you and your techs that you MUST wear these items, they have less liability if someone does get something in their eye.
Example A: Occupational safety does not say to wear the eye shields and Technician A gets abrasive slurry compound in his/her eye. Depending on how that compound affects the eye, he/she is now getting paid comp/injury leave and possibly preparing a lawsuit against the company for not providing a safe work place while OSHA is looking for answers and handing out fines as well.
Example B: Occupational safety says to wear the eye shields and Technician A does not because its an "inconvenience". Technician A gets compound in his/her eye. The company can now say, "Sorry but you were negligent in following proper protocol." The company's management and insurance company can rest easy knowing that any lawsuit has little to no validity due to the negligence of the employee.
I doubt there is much you can do about a perceived safety threat. If you can state that productivity is being hampered due to the implementation of these new safety protocols, management will probably listen to your complaint. What do the directions and MSDSs say for these materials? Do the manufacturers suggest wearing eye shields? I hear Oakley safety spec glasses are expensive but comfortable.
Probably not what you wanted to hear but, for the most part, it's the hard truth. Whether these materials pose a real threat or not, occupational safety views it as a risk and you have to follow the rules. Allowing anyone you supervise to take their glasses off would put your position in jeopardy. Just something to think about.
Chris Zurenko Univ. of Michigan KHRI/Otopathology
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Now that the safety nazis have forced us to wear face shields, smocks } and gloves for handling liquid nitrogen, they want us to wear safety } glasses while doing wedge polishing, which entails frequent looking } through a microscope to judge the progress. The concern is that some } polish slurry 'could' splash into the eyes of a technician. Of course } the safety glasses really get in the way when looking through a } microscope, and our technicians are really not happy about this. I would } like to hear as many opinions as possible on whether there is } significant likelyhood of such a splash, and if polishing slurry, like } Siton or Glanzox really poses a hazard to the eyes of a person using it. } Thanks. } } John Mardinly } Intel
I follow your logic, but it is logic I will resist to the day I retire (which may just have come a little closer!). Possible outcomes for this line of thinking are that many legitimate research activities will effectively be denied us, and more worryingly that this approach to safety is a way for comapnies to deny their responsibilities to employees for their safety. Would you want to work for an employer whose rules effectively abolished any accident insurance. Just something to think about. Chris
Dr. Chris Jeffree University of Edinburgh Biological Sciences EM Facility
----- Original Message ----- } From: "Christopher S. Zurenko" {czurenko-at-umich.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 20, 2003 3:14 AM
In high humidity Michigan, I routinely used an Epon-Araldite receipe from Palevitz and Newcombe (JCB 45(2):383, 1970) that used 40ml DDSA, 20ml Epon (or replacement), 20ml Araldite 6005 and 1.44ml DMP-30. I used a glass syringe for dispensing the DMP-30, a glass rod to stir the resin, de-gassed the resin before use and between resin changes, and kept all components under vacuum. When my resin started behaving oddly I tossed all components and started with fresh.
Good luck!
Angela Welford, EMT Dept of Pathology University of New Mexico Albuquerque, NM, 87131 505-272-1445
The formula should not be changed. We would suggest storing the chemicals in a desiccator cabinet with a desiccant such as silica gel..
JD Arnott
Disclaimer: Ladd Research sells the chemicals and supplies mentioned in this e-mail.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Garry Burgess" {GBurgess-at-exchange.hsc.mb.ca} To: {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 19, 2003 12:25 PM
Garry: I have been using an Epon/Araldite mixture here in Houston for at least the past 15 years with no problems. I doubt that you are any more hot and humid than I am here. I do not measure mine by weight but by volume. I use the following formula: 12 ml of Epon 812 (any substitute 812) 10 ml Araldite 502 24 ml DDSA 1 ml DMP 30
I do all of my measurements using a disposable plastic syringe. I remove the plunger and pour in an amount just over what I need (be sure to keep the tip cover on or it will flow out while you are pouring). After filling the syringe with slightly more than the desired amount, I carefully replace the plunger and invert the syringe, remove the cap and expel the air. Then I discard the small amount over what I need and then expel the measured portion into a disposable 50 ml polypropylene centrifuge tube. Brand does not matter as long as it has a sealable top (I am currently using Falcon Bluemax 2098). I use the same syringe for all of the first 3 components. I measure the DMP30 by placing the tip of a 2 or 3 ml syringe in the bottle and drawing up the desired amount into the syringe. After all the components have been place in the centrifuge tube, I mix by inversion on a rotator for at least 15 minutes. I know many people say you shouldn't mix by inversion because of the air bubbles but I find this is not a problem if I let the mix set for 15 to 20 minutes after mixing. My embedding protocol is as follows 50% ethanol for 15 minutes 70% ethanol for 15 minutes 95% ethanol for 15 minutes 100% ethanol for 15 minutes X2 100% propylene oxide for 15 minutes X2 1:1 Epon/araldite:propylene oxide for 1 hour Pure Epon/Araldite for 1 hour Place one piece of tissue on fresh plastic in a beem capsule and let it slowly settle to the bottom of the capsule. After 3 to 4 hours place in oven at 80o C overnight to polymerize. If you need more info you can email me at msteglic-at-mdanderson.org.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center Houston, TX
Steve, Karen, who was Dr. Keith Porter's Research Specialist, was in my department several years back. They were studing fish scale cells growing on gold grids that were coated but I do not recall with which plastic. The grids were carbon coated indirectly (a shield was placed between the rods and the grids). I do know that the grids were glow discharged while still on the coverslip which was used to pick up the coated grids. A thin strip of aluminum was used instead of the usual copper wire for the glow. The grids were UV sterilized and I believe they were put directly into the culture dishes with no washing/rinsing. They did use 400 kV. to see more than just the edge of the cells. I believe that this work was published before Dr. Porter passed away. Pat
Patricia Stranen Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
} Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (ssamuelsson-at-eyetk.com) from } http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html } on Monday, August 18, 2003 } --------------------------------------------------------------------------- } Email: ssamuelsson-at-eyetk.com } Name: Steve Samuelsson } Organization: Eyetech Pharmaceuticals } Title-Subject: Whole Mounts } Question: Regarding growing endothelial cells on nickel grids for } whole mount TEM analysis: our luck is inconsistent and we now need } to dig further for the reason. 200 and 300 mesh nickel grids are } coated with ~0.5% formvar in ethylene dichloride (we also tried } chloroform) and picked up on a 13mm cover slip. These are allowed } to dry and age a day or longer. Prior to plating, the slips are UV } sterilized 10-15', incubated in 1% gelatin ~45', washed 1x with PBS, } culture medium added followed by cells. Our issue is incomplete (or } downright lousy) growth of cells on the grids. Cells are robust and } growing fine on the bottom of the 24 well plate so we suspect } something with the grids. My first thought was that we needed } carbon coating of the grids but have some trouble with this } explanation. Second thought was that the solvent was still coming } out of the films and poisoning the cells; thus the 'aging' issue } with the cover slips. Any suggestions? } ---------------------------------------------------------------------------
This has happened at our institution also, but for short-sighted folk like me who have to wear glasses on the microscope anyway, "they" made a real effort to get reasonably comfortable and not too large fully enclosed safety glasses with prescription lenses - must have cost a packet. They do have to worry about insurance, etc, but here, at least, although it's an annoyance, it's not too hard to follow the regulations.
} } Now that the safety nazis have forced us to wear face shields, smocks and } gloves for handling liquid nitrogen, they want us to wear safety glasses } while doing wedge polishing, which entails frequent looking through a } microscope to judge the progress. The concern is that some polish slurry } 'could' splash into the eyes of a technician. Of course the safety glasses } really get in the way when looking through a microscope, and our } technicians are really not happy about this. I would like to hear as many } opinions as possible on whether there is significant likelyhood of such a } splash, and if polishing slurry, like Siton or Glanzox really poses a } hazard to the eyes of a person using it. Thanks. } } John Mardinly } Intel
Dr Rosemary White rosemary.white-at-csiro.au Microscopy Centre fax 61- 2 6246 5000 CSIRO Plant Industry ph. 61- 2 6246 5475 or GPO Box 1600 mob. 61- 0402 835 973 Canberra, ACT 2601, Australia
Years ago I grew crustacean neurosecretory cells on Formvar-coated gold grids, fixed, postfixed, dehydrated, and critical point dried them (we had a grid holder for our Tousimis CPD) for 400 kV TEM. They were pretty happy cells, and it worked well.
Aloha, Tina
**************************************************************************** * Tina (Weatherby) Carvalho * tina-at-pbrc.hawaii.edu * * Biological Electron Microscope Facility * (808) 956-6251 * * University of Hawaii at Manoa * http://www.pbrc.hawaii.edu/bemf* ****************************************************************************
Being in the US university system for ten years now (wow, time is just flying) I was and is amused how this system handles safety issue. Students and most technical stuff have very little idea how to perform safe in the laboratory environment. Safety issue is limited to some bureaucratic tasks not related to the real safety (mostly keeping paperwork in a good shape and accurately fill out chemical waste disposal tags). So, in such situation, I think, it's superviser's personal responsibility to provide safely environment to the workers and train them accordingly. I think, the health issue is much, much more important than any personal or corporative ambitious. If it's SAFER to use protective glasses, it's your responsibility to ensure that your workers do use glasses. It's really sad, but I agree at some degree with Chris Zurenko, that safety issue sometime reflects not real carry about worker's health but attempt to avoid legal responsibility (which, in my point of view, nothing to do with real safety). Sergey
P.S. By the way: in my EM work, my glasses, I wear to correct my vision, save my eyes in uncounted number of times. I, also, used to remove them every time I am using light microscope or binoculars. I don't see big problem removing them if necessary. But: yes, it decreases my productivity. Ok, let say, I need about 5 sec. to remove them and put back. Let say I do it 40 times a day: 5 secx40=200 sec/3.3 min per day.
At 12:36 AM 8/20/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
I'm not saying that I follow my own logic 100%. There are tasks that I perform in the lab everyday that I probably "should" be wearing more protective gear, but I don't, and I'm sure we all do. With regard to the original post, I'm not sure that I would want to wear a full smock for pouring liquid nitrogen either, but gloves and eye protection are a good idea.
The point I'm trying to make is that in a day and age of fast food chains getting sued because their fatty food makes people overweight and cigarette companies handing out huge settlements because people now have cancer, etc. (after doctors have been stating for YEARS that fatty food and cigarettes are not good for you), companies can not be too careful. That is the case here. The company implementing the eye guards does not want someone to get abrasive compound in their eyes. They are attempting to limit their risk and liability for someone being injured, very simple. You would like to think that the company has acted on more of a humanitarian agenda to provide for and protect its employees (and it may be a small piece of the pie) but I think it's more to protect the company.
I don't see how legitimate research would be denied due to the implementation of safety rules. There should be rules to limit risk, but you can never 100% eliminate it. Legitimate research grinds on everyday (following internal, local, state and federal safety mandates) and I don't see that ending anytime soon.
Back to the original post - if using this eyewear is creating a major problem for the type of research being conducted, talk with management and the safety people to attempt to implement a mutually beneficial setup where an eye splash could mostly be eliminated. Maybe some sort of Lexan guards or shielding rather than eyewear. As far as the specific question about the abrasive getting in the eye, I don't know but I don't think that matters (though I can't imagine any sort of abrasive in the eye would ever be a good or tolerable thing). It probably wouldn't matter if it's abrasive, oil, sewage, tap water or sterile saline that possesses a risk of getting slashed in the eye. Your safety people have identified a safety risk and have moved to protect the employees and company. Whether you or anyone else agrees with me or not, it is now the responsibility of techs to wear the glasses and supervisors to make sure they are worn, whether anyone is happy about it or not.
Cheers
-------------------------------------------------------------------- Christopher S. Zurenko Research Assistant II Kresge Hearing Research Institute, Otopathology The University of Michigan Medical School MSRB 3, Room 9303 1150 W. Medical Center Dr. Ann Arbor, MI 48109-0648
Garry: Please check to storing conditions of your components, the DDSA and less the DMP 30 are very hidroscopyc. The last one can be replaced with BDMA with very good results at ultramicrotomy time. Have a good luck!
Francisco Freire FRMS Head Electron Microscopy Service University of Las Palmas de Gran Canaria Canary Islands Spain
In my labs it is mandatory to wear safety glasses. Risk assessments are carried out on all microscope usage and, if deemed safe, can be given an exemption sticker (signed off by management) so the user can remove specs. That RA would cover what activities are occurring adjacent and whether the operative has wet hands/gloves as any contaminants could be transferred to hair/skin and indeed eyes during removal of the glasses.
Where the 'environment' remains unsafe we are now moving to replacing the eyepieces with those suitable for spectacle wearers (safety or not). Non spectacle users in Histopath labs here are also having them fitted for doing their 'safe' microscopy as they prefer the ergonomics of them.
So change the microscopy set up and every-one should be safe and happy.
Gillian Brown Histology Section, Asthma Biology RI CEDD, GSK.
After a massive (or at least sobig) virus attack I'm back online.
That's a very interesting point Sergej makes.
We work with thin (1 layer) protein crystals and large individual protein molecules (chaperones etc.).
For crystals the flatness of the carbon surface is obviously important but for single proteins I've always thought it shouldn't make any difference. Initially we use negative stain to look at the quality of a specimen. To get higher resolution data we use cryo-specimens on holey carbon so that is a different story. Back to the negative stain: I've been wondering about two possible effects when staining: 1. When you stain a specimen on the rough side the stain might fill all the valleys of the carbon surface leading to large background noise and making it difficult to see the particles (some grids seem to have a lot of background). 2. When you stain a specimen on the smooth side (which is facing the grid) the stain might get trapped between the grid bars leading to an unnecessarily thick layer of stain which again increases the noise.
Are either of the two real problems?
Philip Koeck Svdertvrns Hvgskola and Karolinska Institutet Dept. of Bioscience at Novum S-14157 Huddinge Sweden phone: +46-8-6089186 fax: +46-8-6089290 http://www.biosci.ki.se/em _______________________________________
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Monday, August 18, 2003 10:06 PM
The attachments with .pif extensions
This is the virus you may have. http://securityresponse.symantec.com/avcenter/venc/data/w32.sobig.f-at-mm.html
Note that the virus is spoofing e-mail addresses. so your system could be clean.
Pavel Lozovyy ATC SEM Lab Phone: 216-692-6637 E-mail: atclabs-at-sbcglobal.net web: www.atclabs.com
first, i am not really sure what the position of chris jeffree is, i found his response a bit confusing. sergey's comments considering time are to the point, and rosemary describes the situation i have experienced quite accurately. from my view, after thirty years i still have my eyes. last time i checked, they were a bit important for my work.
in a previous persona (before i grew up and went to graduate school) i had the privilege of negotiating collective agreements on behalf of an independent union which represents university non-academic staff. at the university of manitoba this means about 2000 administrative, clerical, computer, library, security and technical employees. the technical staff were, and still are about 40% of the population. for the record, i was chief negotiator, not member of the team, and in one of those cases we were on strike for 7 weeks. hopefully this sets it clear with chris zurenko that i was not a management stooge. the issue of safety supplies and equipment was always important, even in the dark ages of 1975. in fact, historically most improvements in working conditions and staff safety are the result of labour organizations fighting for them. five day work weeks, vacations, and child labour laws were not the result of an insurance company telling managers they had to make the changes. they were the result of the workers fighting for better and safer working conditions. in our case, we identified problems long before any insurer or government agency ever did. what is most important is that the university of manitoba, at least, never argued the importance of the issues. usually, our only problem was figuring out how we could ensure payment for the needed materials in individual grant funded positions.
having worked in the university research system for over 30 years i have seen most different approaches. to make it clear, i do not think universities are any more concerned with safety than any employer. have the 'safety nazis' made it more difficult, yes. but at least the safety issues are being dealt with in many areas, not by a few benevolent employers. no insurance payment will compensate for the use of a hand, hearing, life, or in most of our cases, an eye. if you have a problem with implimentation do what rosemary did, discuss the methods, rationally. do it as if you were defending your thesis again. sell the problem and get a cure. but let's listen to sergey and not complain about the few seconds needed to take off glasses or shields.
finally, to make it clear, i learned my attitudes from my manufacturer father, who taught me that the most important resource he had was his workers. i am not a left wing pinko, i am very middle of the road. and perhaps a little to the right of centre on economic issues
paul
Paul R. Hazelton, PhD Electron Microscope Unit University of Manitoba Department of Medical Microbiology 531 Basic Medical Sciences Building 730 William Avenue Winnipeg, Manitoba, Canada, R3E 0W3 e-mail: paul_hazelton-at-umanitoba.ca Phone:204-789-3313 Pager:204-931-954 Cell:204-781-1502 Fax:204-789-3926
Can anyone recommend a microscopy method/technique to visualize and/or possibly quantify the amount of free plasticizer vs the amount of bound plasticizer in cast PVC skins? We suspect a ratio difference of free vs bound plasticizer when comparing PVC raw materials vs cast PVC skins which are exhibiting adhesive failure between the PVC skin and foam. We think there may be a higher amount than normal, of free plasticizer migrating to the PVC skin surface.
Fred A. Hayes Polymer Microscopy Collins & Aikman IntelliMold Div 4651 Platt Lane Ann Arbor, MI 48108 734-477-7029 office 734-477-9214 fax
Only for windows. Mac users never have the problem. :) I received about 30..opened some..no problem. :)
on 8/21/03 7:24 AM, "atcsem-at-earthlink.net"-at-sparc5.microscopy.com at "atcsem-at-earthlink.net"-at-sparc5.microscopy.com wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } The attachments with .pif extensions } } This is the virus you may have. } http://securityresponse.symantec.com/avcenter/venc/data/w32.sobig.f-at-mm.html } } Note that the virus is spoofing e-mail addresses. so your system could be } clean. } } Pavel Lozovyy } ATC SEM Lab } Phone: 216-692-6637 } E-mail: atclabs-at-sbcglobal.net } web: www.atclabs.com } } }
In Europe (I am not sure of the USA) there is a general system of controlling exposure to hazards which involves in order of preference: 1.Prevent exposure (eg remove the hazardous process or substitute hazardous material with safer one). 2.Control process by design/engineering (eg shielding, fume hood). 3.Use procedural controls (eg less people involved, good hygiene, reduce quantities). 4.and only finally use PPE (personal protective equipment) such as safety glasses.
If the first three can be applied successfully then there should be no need for PPE such as safety glasses. An obvious point may be to separate the preparation procedure from the microscopy because if there is a risk of splashing the eyes then surely there is a risk of splashing the optics of microscopes.
It should also be borne in mind that if an accident (at least in the UK) results in an injury and the employer didn't take reasonable precautions and enforce them then that employer could be deemed to be negligent. Health and safety issues are often covered by criminal law so the employer or their representative could go to prison (doesn't often happen - but can) and any damages would not normally be insurable so the employer would have to pay and not the insurer. It gives a different slant on the "safety nazis" .
I am still a union health and safety officer and so my sympathies are with the staff rather than employers but I do recognize how difficult a job employers can sometimes have.
I'm sorry for extending an already lengthy debate but the principles are extremely important and I have tried to be brief.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: "paul r hazelton, PhD" {paul_hazelton-at-umanitoba.ca}
Sergey! You say that you can't use the bar side of the grids. Why is that so clear? A thicker layer of stain might not be a disadvantage as long as it is even.
(I'm assuming specimens that are much smaller than the mesh)
Philip
} Phillip - you right: on the rough surface negative stain fills the } imperfections of the surface and therefore increase background noise. It } also makes stain's layer thicker (holding more stain then necessary). All } these things decrease the resolution of your images. You could not use, of } coarse, the opposite side of the grid with bars. So, you need to put flat } side of the carbon up: mount your grids ON the floating carbon. If you are } trying to do high resolution EM on individual proteins, you definitely need } to pay attention to the quality of the carbon: this is a whole point to } use "mica-side" of the carbon, because mica surface is atomically perfect } (it's a single crystal), so the carbon surface will be nearly perfect as } well. The quality of mica is important too. Have a good day, Sergey. } } } } } } } } After a massive (or at least sobig) virus attack I'm back online. } } } } That's a very interesting point Sergej makes. } } } } We work with thin (1 layer) protein crystals and large individual protein } } molecules (chaperones etc.). } } } } For crystals the flatness of the carbon surface is obviously important } } but for single proteins I've always thought it shouldn't make any } } difference. } } Initially we use negative stain to look at the quality of a specimen. } } To get higher resolution data we use cryo-specimens on holey carbon so } } that is a different story. } } Back to the negative stain: I've been wondering about two possible effects } } when staining: } } 1. When you stain a specimen on the rough side the stain might fill all the } } valleys } } of the carbon surface leading to large background noise and making it } } difficult } } to see the particles (some grids seem to have a lot of background). } } 2. When you stain a specimen on the smooth side (which is facing the grid) } } the stain } } might get trapped between the grid bars leading to an unnecessarily thick } } layer of } } stain which again increases the noise. } } } } Are either of the two real problems? } } } } } } } } } } Phillip } } } Your way to mount carbon on the grids has one very serious disadvantage: } } } } } when you mount carbon this way, you will use "air-side" of carbon as a } } } working surface. This side is usually more grainy and dirty. The whole } } } idea of using carbon from mica is to use "mica-side" of carbon as a } } working } } } surface. The idea here is that the carbon face toward mica is more } } uniform } } } and perfectly clean (it was not exposured to the air). In order to use } } } "good" side of carbon, you have to put grids on floated carbon and then } } } pick the grids with Parafilm for instance. Similarly, you need to do the } } } same for plastic film created on water - working side is "water-side", not } } } air. Only one exception I do know is Formvar film created on the } } } glass. Glass is not such clean and perfect as mica or water surface, } } } therefore, you need to use "air-side". Then your technique will work } } } perfectly. I also use your technique to mount holey film on the grids. } } } Best wishes, Sergey. } } } } } } } } } } } } } } } } } } You can easily use the method described by Mike to make } } } } about 50 pure carbon grids at a time (much cheaper than } } } } buying if you already have the equipment). } } } } } } } } You place about 50 grids on a square piece of filter paper } } } } lying on a little "table" made of wire mesh under the water surface. } } } } To get the grids under water make them wet first and push them } } } } through the surface vertically. } } } } Then you lower the water surface by removing water from underneath } } } } (for example with a syringe or a plastic tube with a valve) to place } } } } the carbon film (floated off the mica) on top of the grids. } } } } (Of course the carbon film has to be in one piece in this case.) } } } } Push the carbon into position with something blunt (a paddle ? :) } } } } } } } } Philip Koeck
} } } } } } Steve, } } } } } } When I want just carbon as a substrate I will } } } } } } coat freshly cleaved mica with carbon in a metal } } } } } } evaporator. This allows me to dictate the carbon } } } } } } thickness and I have no trouble floating off grid } } } } } } size squares(prescored)in a drop of water. Picking up the } } } } } } carbon with glow discharged grids (flat side) } } } } } } is a snap. Make sure everything is clean. } } } } } } This should give you thin clean substrates. } } } } } } Mike D. } } } } } } }
You mentioned a good point with regard to chemistry and microscopy, wet gloves. Many people do etching, polishing, and wet chemistry with iterations between a hood/sink and a microscope. Some also feel that they are saving time/money by not removing their gloves whilst adjusting the focus, stage position and worst of all the eyepieces. I can say from experience how this caused me great pain and an emergency hospital visit because someone had their gloved hands in nitric acid and decided to adjust the inter-ocular distance on a microscope that I was alternately using.
My rule was that no gloved hand may touch a microscope whether it's been in a hood, sink or whatever. This way there can be no mistake unless some fool likes to work with bare skin, and there is no protecting oneself from the super-idiot. The only remedy for them is banishment to a desk job where the limit of injury is paper cuts or burns from hot coffee.
Peter Tomic Agere Systems Allentown, PA
-----Original Message----- } From: "gillian.2.brown-at-gsk.com"-at-sparc5.microscopy.com [mailto:"gillian.2.brown-at-gsk.com"-at-sparc5.microscopy.com] Sent: Thursday, August 21, 2003 5:46 AM To: microscopy-at-sparc5.microscopy.com
In my labs it is mandatory to wear safety glasses. Risk assessments are carried out on all microscope usage and, if deemed safe, can be given an exemption sticker (signed off by management) so the user can remove specs. That RA would cover what activities are occurring adjacent and whether the operative has wet hands/gloves as any contaminants could be transferred to hair/skin and indeed eyes during removal of the glasses.
Where the 'environment' remains unsafe we are now moving to replacing the eyepieces with those suitable for spectacle wearers (safety or not). Non spectacle users in Histopath labs here are also having them fitted for doing their 'safe' microscopy as they prefer the ergonomics of them.
So change the microscopy set up and every-one should be safe and happy.
Gillian Brown Histology Section, Asthma Biology RI CEDD, GSK.
your points are very good. i don't know that i have seen them written down here, but they fit exactly the process we use here. there is heavy emphasis on point three. procedure controls. if a person is only going to do something only 1x or 2x, we do it for them. this does not matter if it is dealing with pathogens, radiation, osmium, or other chemical or biologicals.
for what it is worth, i have been trying to eliminate all use of radioactives from my work - not because of the necessary controls and protection, but because of the paper work, that unmentioned administrative point, #5.
thanks for the points, i will forward them to my former union.
I see you've already had a detailed explanation (which I've lost - accidentally deleted because of something odd in my software - never mind, I can pick it up again once it's in the archive), however, I would like to share with you some experience of observing polymer spherulites under the polarizing microscope.
Normally, one observes spherulites between crossed polars, and if the birefringence is strong, you get a four lobed pattern with colours in the sequence you also observe with the quartz wedge. However, if you rotate the analyser to be parallel with the polarizer, you see different colours, in a two lobe pattern. These colours are the complementary ones to those observed between crossed polars. Going from no birefringence to just over the 1st order, one has:
Crossed - Parallel
Black - White (zero order) Grey - not so bright Yellow - Dark Blue Purple - Green (1st order) Blue - Reddish Orange
I have often looked at polymers (including blends) under the polarizing optical microscope. I do not find that rotating the analyser gives extra information, it simply changes the colour of the presentation. However, rotating the specimen between crossed polars, and watching the colours brighten and darken (though they remain the same colour) does give information about the orientation of the molecules.
Hope this is of use.
+-----------------------------------------+ Robert H.Olley J.J.Thomson Physical Laboratory University of Reading Whiteknights Reading RG6 6AF England +----------------------------------------+ Phone:+44 (0) 118 9318572 Fax: +44 (0) 118 9750203 University internal extension 7867 Email: R.H.Olley-at-reading.ac.uk URL: http://www.reading.ac.uk/~spsolley +----------------------------------------+
My company is in the semiconductor industry. We current have an AMRAY 2030 C for performing defect review on 8-inch wafers with the ability to read defect files from wafer inspection tools. Our AMRAY is on its last legs.
Are there lab SEM available that can inspect 8-inch wafers edge to edge with the ability to read defect files?
The semiconductor SEM's are expensive and restrictive for r&d work. They are design for production purposes and many SEM controls are removed or automated that limit the tools usefulness.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (yezer-at-barak-online.net) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Saturday, August 23, 2003 at 10:16:49 ---------------------------------------------------------------------------
Email: yezer-at-barak-online.net Name: Ezer Yosi
Title-Subject: MListserver:
Question: I am looking for a fast glue which is compatible with UHV (for TEM sample). Can you kindly recomend on one of loctite system for this application.
Ezer Yossef wrote: ===================================================== Question: I am looking for a fast glue which is compatible with UHV (for TEM sample). Can you kindly recomend on one of loctite system for this application. --------------------------------------------------------------------------- The M-Bond 610 system seems to be the most widely used glue for this application, and it can be purchased from SPI Supplies (or some of the other major supply houses for EM consumables) and is described on URL http://www.2spi.com/catalog/spec_prep/glue.shtml
I do hear reports of some who claim to get comparable results with some of the "5 minute" epoxy systems, such as is shown on URL http://www.2spi.com/catalog/spec_prep/5minglue.shtml
The latter is obviously much cheaper but most people seem to prefer M-Bond 610 (or possibly other alternatives that might be comparable to M-Bond 610).
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
One more thing add under your message. The Mo grid with very smooth hole will provide the most flat carbon film without cryo-wrink. For more details, please cheak Pacific Grid-Tech website.
http://www.grid-tech.com/
regards
Wendy
} -----------------------------------------------------------------------. } } } After a massive (or at least sobig) virus attack I'm } back online. } } That's a very interesting point Sergej makes. } } We work with thin (1 layer) protein crystals and } large individual protein } molecules (chaperones etc.). } } For crystals the flatness of the carbon surface is } obviously important } but for single proteins I've always thought it } shouldn't make any } difference. } Initially we use negative stain to look at the } quality of a specimen. } To get higher resolution data we use cryo-specimens } on holey carbon so } that is a different story. } Back to the negative stain: I've been wondering } about two possible effects } when staining: } 1. When you stain a specimen on the rough side the } stain might fill all the } valleys } of the carbon surface leading to large background } noise and making it } difficult } to see the particles (some grids seem to have a lot } of background). } 2. When you stain a specimen on the smooth side } (which is facing the grid) } the stain } might get trapped between the grid bars leading to } an unnecessarily thick } layer of } stain which again increases the noise. } } Are either of the two real problems? } } } Philip Koeck } Svdertvrns Hvgskola and } Karolinska Institutet } Dept. of Bioscience at Novum } S-14157 Huddinge } Sweden } phone: +46-8-6089186 } fax: +46-8-6089290 } http://www.biosci.ki.se/em } _______________________________________ } } } ----- Original Message ----- } } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} } To: {microscopy-at-sparc5.microscopy.com} } Sent: Monday, August 18, 2003 10:06 PM } Subject: Re: Carbon Support Films for TEM } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Phillip } } Your way to mount carbon on the grids has one very } serious disadvantage: } } } when you mount carbon this way, you will use } "air-side" of carbon as a } } working surface. This side is usually more grainy } and dirty. The whole } } idea of using carbon from mica is to use } "mica-side" of carbon as a } working } } surface. The idea here is that the carbon face } toward mica is more } uniform } } and perfectly clean (it was not exposured to the } air). In order to use } } "good" side of carbon, you have to put grids on } floated carbon and then } } pick the grids with Parafilm for instance. } Similarly, you need to do the } } same for plastic film created on water - working } side is "water-side", not } } air. Only one exception I do know is Formvar film } created on the } } glass. Glass is not such clean and perfect as } mica or water surface, } } therefore, you need to use "air-side". Then your } technique will work } } perfectly. I also use your technique to mount } holey film on the grids. } } Best wishes, Sergey. } } } } } } At 04:07 PM 8/18/03 +0200, you wrote: } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The } Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } } -----------------------------------------------------------------------. } } } } } } } } } You can easily use the method described by Mike } to make } } } about 50 pure carbon grids at a time (much } cheaper than } } } buying if you already have the equipment). } } } } } } You place about 50 grids on a square piece of } filter paper } } } lying on a little "table" made of wire mesh under } the water surface. } } } To get the grids under water make them wet first } and push them } } } through the surface vertically. } } } Then you lower the water surface by removing } water from underneath } } } (for example with a syringe or a plastic tube } with a valve) to place } } } the carbon film (floated off the mica) on top of } the grids. } } } (Of course the carbon film has to be in one piece } in this case.) } } } Push the carbon into position with something } blunt (a paddle ? :) } } } } } } Philip Koeck } } } Svdertvrns Hvgskola and } } } Karolinska Institutet } } } Dept. of Bioscience at Novum } } } S-14157 Huddinge } } } Sweden } } } phone: +46-8-6089186 } } } fax: +46-8-6089290 } } } http://www.biosci.ki.se/em } } } _______________________________________ } } } } } } } } Steve, } } } } } When I want just carbon as a substrate I } will } } } } } coat freshly cleaved mica with carbon in a } metal } } } } } evaporator. This allows me to dictate the } carbon } } } } } thickness and I have no trouble floating off } grid } } } } } size squares(prescored)in a drop of water. } Picking up the } } } } } carbon with glow discharged grids (flat side) } } } } } is a snap. Make sure everything is clean. } } } } } This should give you thin clean substrates. } } } } } Mike D. } } } } } }
===== Pacific GridTech A high quality EM grid provider 3505 Caminito Carmel Landing San Diego, CA 92130, USA Tel: (858) 336 8938; Fax: (858) 259 5511 Email: info-at-grid-tech.com Web: http://www.Grid-Tech.com/
First off, let me comment about UHV and TEM. UHV is generally considered to be in the vacuum range where the pressure is less than 1E-9 Torr. Although the microscope manufacturers would like you to believe that your sample is setting in an UHV environment, chances are it isn't. The dedicated STEM systems may come close. The reason for this is that the pressure is typically measured using the ion pump or a gauge located close to the pump flange. Your sample sits at the end of a conductance limited tube that connects to your pump. In vacuum systems, the throughput (quantity of gas per unit time), Q (Torr l/sec) = S * P, where S is the pump speed (l/sec) and P is pressure (Torr). S * P at the pump is equal to S(eff)*P(sample). S(eff) is the effective pump speed at your sample and is related to both the conductance and pump speed. However, if your diameter of the tubing is small and the length is long or has bends in it, the effective pump speed will be conductance limited and is approximately the same as the conductance. The net effect is that the effective pump speed at your sample is always less than the pump speed and as a result, the true pressure at your sample will be higher than the reported pressure. With all that said, if you cool an anti-contamination device in the neighborhood of your sample, that acts as a cryopump and will lower the pressure at your sample.
For high vacuum systems, epoxy based materials will work fine. However, you should make sure that the mix is right so that everything cross links well and you don't have any of either the hardener or the resin "left over". You should also minimize the amount that you use. Epoxies that have worked well in TEM sample prep are M-bond 610 (as Chuck Garber said), Gatan's G-1 which is equivalent to Epoxy Technologies' Epo-Tek 353ND. A silver filled conductive epoxy that works well is Epoxy Technologies' H-22. Instead of the 5 or 10 min epoxies, I would recommend the 90 min working time, super strength types that require an overnight cure.
If your interested in true, UHV compatible "glues", you are in trouble. There are two sealers that people use for this purpose in UHV systems. One is a product called VacSeal(R) and comes in a brush-on bottle form or a spray application. This stuff works very well when it is room temperature dried, but works even better with a 200C cure. I doubt that it would work well for TEM sample prep. Another material that is used is a two part putty-like material that used to be available from Perkin Elmer. Sorry, I can't remember the name. I would check with vacuum component manufacturers for the name. You might also check with Kurt Lesker. People have also used the water based "Aquadawg" or "Aquadog" to tack things down in UHV systems, but it has to be heated to drive off the water.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
We are in the market for an EBSD system. (I believe all the relevant vendors already know this.) This is a very new area for us and we have basically no experience in this area (to be very blunt). We are dealing with technical specs, vendor information, and demos. And as good and as hands- on as demos maybe they can not give you the true picture of working with a system for months or years or multiple users. What we are looking for is information from USERS of EBSD systems (Researchers, facility managers, etc.) - we are looking for comments, opinions, pro & cons, experiences of any EBSD systems.
Please send your comments directly to me so you maybe as candid as possible.
Thank you.
Richard E. Edelmann, Ph.D. Electron Microscopy Facility Supervisor 350 Pearson Hall Miami University, Oxford, OH 45056 Ph: 513.529.5712 Fax: 513.529.4243 E-mail: edelmare-at-muohio.edu http://www.emf.muohio.edu
I'd suggest you investigate both Raman confocal and FT-IR microscopy.
The FT-IR is probably easier to do, if you can get cross sections. Otherwise, Raman confocal can go down layer by layer. I don't think that there is any limitation for either system in terms of the chemical fingerprint they can provide to you. The bound v. unbound may have a better signature in Raman, however.
A number of companies, including BioRad and Nicolet, provide FT-IR microscopes. SensIR (Danbury, CT) provides the IlluminatIR - a unique microscopy accessory which turns a research microscope into an FT-IR microprobe without disabling the microscope.
Renishaw and Jobin-Yvon (JY) both provide Raman confocal.
All of the companies have apps labs which can run a sample or two for you. I'd suggest you visit microscopy.info for contact information. I'm going to be on vacation most of this week, but if you get stuck, drop me an email and I'll send you more specific contact information.
Good hunting! Barbara Foster Microscopy/Microscopy Education 125 Paridon Street, Suite 102 Springfield, MA 01118 PH: 413-746-6931 FX: 413-746-9311 Web: www.MicroscopyEducation.com
^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& Optimizing Light Microscopy is still available through MME. For information on ordering single or small quantities, visit our website. For discounts on classroom-sized, call us today. ^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^&^& At 11:19 AM 8/21/03 -0400, Fred.Hayes-at-colaik.com"-at-sparc5.microscopy.com wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
(Please do not respond to this e-mail. Send all responses to the contact listed below)
A position is available in the Materials Science and Engineering Department at Carnegie Mellon University (http://neon.mems.cmu.edu/newfront.shtml) for a full time staff member with experience in transmission electron microscopy. The position will focus on analysis of ceramic thin films having applications in the areas of electronics, magnetics, hard coatings, and catalysis. A significant portion will focus on nano-layered thin films for use as hard coatings. Tasks will include sample preparation (of plan-view and cross-sectional samples), electron microscopy (including high-resolution), and analysis of structural details (such as crystal, interfacial, grain, and superlattice structures). The position could be filled either at the research scientist or research associate (postdoctoral) level. Salary will be commensurate with experience. Carnegie Mellon University is an equal opportunity employer.
For Details and Inquiries Please Contact Paul Salvador Assistant Professor Department of Materials Science and Engineering Wean Hall 3301 Carnegie Mellon University Pittsburgh, PA 15206 TEL: 412-268-2702, FAX: 412-268-7596 paulsalvador-at-cmu.edu
To all those people out there who use the Gatan PIPS -
I thought this was common knowledge, but maybe not. Instead of polishing the sputtered deposits off the viewing window after every hour or so of milling time, just use a grade 0, 19 mm diameter glass cover slip. It sits nicely in the recess under the window - the only precaution you have to take is to make sure it doesn't slide to partially cover the O-ring. They are cheap and can be considered disposable, but if you are really tight and have a big jar of KOH in the cupboard (like me) you can clean the deposits off by boiling in caustic solution for a while and re-use them.
I just hate to think of all those cumulative hours spent by users around the world polishing glass when there is a simple alternative!
======================================================================= This e-mail is intended for the person it is addressed to only. The information contained in it may be confidential and/or protected by law. If you are not the intended recipient of this message, you must not make any use of this information, or copy or show it to any person. Please contact us immediately to tell us that you have received this e-mail, and return the original to us. Any use, forwarding, printing or copying of this message is strictly prohibited. No part of this message can be considered a request for goods or services. ======================================================================= Any questions about Bookham's E-Mail service should be directed to postmaster-at-bookham.com.
I would like to try poly-L-lysine for sticking cells to coverslips etc for dehydration and SEM. My S**** catalogue offers about 15 variants. Can anyone advise me on which type to use?
Dave
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
I'm looking for any feedback on fixation of eyes (small and large animal). More specifically: a fixative that would work well for Histology and EM. Any info. on: * Glutaraldehyde * Karnovsky's Fixative * McDowell - Trump's Fixative or any other fixative would be greatly appreciated.
Thanks in advance,
Pedro
********************************************************************* This message and any attachments are solely for the intended recipient. If you are not the intended recipient, disclosure, copying, use or distribution of the information included in this message is prohibited -- Please immediately and permanently delete.
The 2-part epoxy is called Torr-seal, and is is very good. --John Mardinly
material that is used is a two part putty-like material that used to be available from Perkin Elmer. Sorry, I can't remember the name. I would check with vacuum component manufacturers for the name. You might also check with Kurt Lesker. -Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
David There is some trick with Poly-Lysine solution. I would suggest to use some from EM major suppliers. Personally I am using 1% solution from TedPella. I don't think there is a difference between different brands. In England you may contact Agar, very good EM supplier. Good luck, Sergey.
At 07:11 AM 8/26/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Dear Listers, I am looking for a lab that can perform TEM analysis of high impact polystyrene. I intend to obtain an extended support for next few months in order to analyze 30-35 samples. The analysis involves specimen preparation (cryomicrotomy, staining), imaging at 2-3 reasonable magnifications and calculation of rubber volume fraction/size of each sample. I will appreciate the help of the list members in this regard. The interested labs on the list server may want to contact me of line for further discussions regarding the project.
Best regards.
Toseef Ahmed, Ph.D. Electron Microscopy and Thermal Labs, Analytical Section SABIC R&T, P.O. Box 42503, Riyadh 11551, Saudi Arabia Tel: (966-1) 265-3333 Ext.6003, 6104 Fax: (966-1) 265-1101 E-mail: toseef-at-sabic.com,
My original post was ambiguous. I am looking for ways to stick fixed cells to coverslips to avoid repeated centrifugation during dehydration for SEM.
Dave
On Tue, 26 Aug 2003 15:11:46 +0100 (GMT Daylight Time) "Patton, David" {David.Patton-at-uwe.ac.uk} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } I would like to try poly-L-lysine for sticking cells to } coverslips etc for dehydration and SEM. My S**** catalogue } offers about 15 variants. Can anyone advise me on which } type to use? } } Dave } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England" } } } } } This incoming email to UWE has been independently scanned for viruses and any virus detected has been removed using McAfee anti-virus software } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Torr-Seal is available from many of the supply houses including Ladd. See http://www.laddresearch.com/General_Catalog/Chapter_11/Miscellaneous_Vacuum_ Accessori/miscellaneous_vacuum_accessori.html for our selection. MSDS is also available on line for those seeking more information.
Debra Sicard Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com ----- Original Message ----- } From: "Mardinly, John" {john.mardinly-at-intel.com} To: "Walck, Scott D." {walck-at-ppg.com} ; "Microscopy (E-mail)" {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, August 26, 2003 1:12 PM
Hi all,
We are a University microscopy recharge center and will be installing a single beam FIB within the next 6 months. I've got to establish a reasonable initial hourly use rate without the benefit of experience with a FIB costs and use patterns. Will you university FIB users out there share some of your experience, wisdom and rates? Off line will be best I think.
Owen
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (maloneyb-at-fiu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Monday, August 25, 2003 at 14:36:09 ---------------------------------------------------------------------------
Email: maloneyb-at-fiu.edu Name: Barb
Organization: FIU
Title-Subject: Phillips 200 TEM
Question: Does anyone have the specs on this particular model? Need ASAP Thanks Barb
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (info-at-klughammer.de) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, August 27, 2003 at 03:42:53 ---------------------------------------------------------------------------
does anybody know a EPI fluorescence microscope manufacturer which offers quite good quality at an affordable price. They will be used for university beginner groups.
This is way off the microscopy topic, but unfortunately, it is relevant to our daily activities of late. I am trying to contact someone probably associated with this list to inform them that they are infected with the SoBig virus and don't appear to know it yet.
Like many of us, I have been receiving dozens of copies of the SoBig virus of late. Being a curious type and being the IT guy for the computers in our labs, I have been poking into some of the messages to see where they came from. Even though the Return Path: and From: fields are usually spoofed, I found that many were coming from a couple of sources. I have included the relevant information below.
Since many of the alleged senders were frequent posters to this list I am guessing that the real sender may be a member of this list. So if BILLPC on SYMET.NET or PRIVAT on T-DIALIN.NET means something to one of you out there, please see about getting your PC disinfected.
Thanks.
Warren
Received: from PRIVAT (pD9E7724F.dip.t-dialin.net [217.231.114.79]) by despam-3.iastate.edu (8.12.4/8.12.4) with ESMTP id h7QCNLKG007344 for {wesaia-at-iastate.edu} ; Tue, 26 Aug 2003 07:23:22 -0500 } From: {martin.voecks-at-web.de}
Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7REWjbX018529 for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 09:32:46 -0500 Message-Id: {200308271432.h7REWjbX018529-at-despam-2.iastate.edu} } From: {gary-at-gaugler.com}
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
We do have the capibiilty. What kind of arrangement would you propose to have the work done?
**************************************** Chaoying Ni, PhD The W.M. Keck Electron Microscope Facility Facility location: 022 Spencer Laboratory Mailing address: 201 duPont Hall Dept of Matls Sci & Eng University of Delaware Newark, DE 19716 (302) 831-8354 (O); -2318(L); -4545(Fax) http://eml.masc.udel.edu *****************************************
On Wed, 27 Aug 2003, AHMED, TOSEEF wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear Listers, } I am looking for a lab that can perform TEM analysis of high impact polystyrene. I intend to obtain an extended support for next few months in order to analyze 30-35 samples. The analysis involves specimen preparation (cryomicrotomy, staining), imaging at 2-3 reasonable magnifications and calculation of rubber volume fraction/size of each sample. I will appreciate the help of the list members in this regard. The interested labs on the list server may want to contact me of line for further discussions regarding the project. } } Best regards. } } } Toseef Ahmed, Ph.D. } Electron Microscopy and Thermal Labs, Analytical Section } SABIC R&T, P.O. Box 42503, Riyadh 11551, Saudi Arabia } Tel: (966-1) 265-3333 Ext.6003, 6104 } Fax: (966-1) 265-1101 } E-mail: toseef-at-sabic.com, } }
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
I've found good success sticking live bacteria cells to the coverslips FOR fixing and dehydrating... Prep (slides or coverslips) Clean slides in Ethanol with a small amount of dilute HCl Rinse in Distilled water Soak for 30 minutes in your Poly-L solution Let air dry (can be stored desiccated for a long time) The Solution that works well and as is published in Steven E. Ruzin's Plant Microtechnique and Microscopy, calls for 100µg-1mg/ml and Tris 10mM, pH 8.0
I've had success with a "thats about enough" diluting the stock Poly-L with Distilled water (one or two mls per 100-200mls of dH20).
This has worked well for general histochemistry (wax sections) as well as liquid suspended bacterial preparations for the SEM. Samples survive fixation, washing, dehydration, and even CPD with significant coverage where the Poly-L is.
We now get our Poly-L-Lysine from the big F chemical/supply company (the S chemical company also has it) the 1% solution is what we get.
HTH
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
} -----Original Message----- } From: Patton, David [mailto:David.Patton-at-uwe.ac.uk] } Sent: Wednesday, August 27, 2003 4:20 AM } To: Microscopy-at-sparc5.microscopy.com } Subject: Poly-L-lysine: clarification } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } My original post was ambiguous. I am looking for ways to } stick fixed cells to coverslips to avoid repeated } centrifugation during dehydration for SEM. } } Dave } } } On Tue, 26 Aug 2003 15:11:46 +0100 (GMT Daylight Time) } "Patton, David" {David.Patton-at-uwe.ac.uk} wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } I would like to try poly-L-lysine for sticking cells to } } coverslips etc for dehydration and SEM. My S**** catalogue } } offers about 15 variants. Can anyone advise me on which } } type to use? } } } } Dave } } } } ---------------------------------------- } } Patton, David } } Email: David.Patton-at-uwe.ac.uk } } "University of the West of England" } } } } } } } } } } This incoming email to UWE has been independently scanned for viruses } and any virus detected has been removed using McAfee anti-virus software } } } } } } ---------------------------------------- } Patton, David } Email: David.Patton-at-uwe.ac.uk } "University of the West of England"
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (pkrsko-at-stevens-tech.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, August 27, 2003 at 12:10:52 ---------------------------------------------------------------------------
Email: pkrsko-at-stevens-tech.edu Name: Peter Krsko
Organization: Stevens Inst Tech
Education: Graduate College
Location: Hoboken, NJ
Question: Hello,
I would like to find out if it's possible to get phosphorus used on TEM screens in solution. I am using simple electron beam mask and I want to highlight the apperture opening with this paint.
Your help is greatly appreciated.
Thank you.
Peter Krsko
P.S: Here are some images for you: http://attila.stevens-tech.edu/~pkrsko/billboart/
Pedro, Karnowsky's is the best. works both for LM and EM. Shashi CCMB Hyderabad INDIA
I'm looking for any feedback on fixation of eyes (small and large animal). More specifically: a fixative that would work well for Histology and EM. Any info. on: * Glutaraldehyde * Karnovsky's Fixative * McDowell - Trump's Fixative or any other fixative would be greatly appreciated.
Thanks in advance,
Pedro
__________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com
Toseef Ahmed, Ph.D. wrote: ======================================================= I am looking for a lab that can perform TEM analysis of high impact polystyrene. I intend to obtain an extended support for next few months in order to analyze 30-35 samples. The analysis involves specimen preparation (cryomicrotomy, staining), imaging at 2-3 reasonable magnifications and calculation of rubber volume fraction/size of each sample. I will appreciate the help of the list members in this regard. The interested labs on the list server may want to contact me of line for further discussions regarding the project. ======================================================= There are a number of independent analytical laboratories in the USA and elsewhere set up to do this type of TEM work for clients. We maintain a listing of some of these laboratories on our website under our "Hot Services" section, see URL http://www.2spi.com/hot-service7.html
Structure Probe, Inc. has been conducting this type of characterization work on high impact polystyrene (HIPS) since the inception of our business in 1970, both cryo and at RT. We are also accredited by A2LA to the standard of ISO Guide 17025. We are not a university, but are a commercial business. Providing a high standard of service to our clients is our #1 priority.
Chuck
=================================================== Charles A. Garber, Ph. D. Ph: 1-(610)-436-5400 President 1-(800)-2424-SPI Structure Probe, Inc. FAX: 1-(610)-436-5755 PO BOX 656 e-mail: cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust. Service: spi2spi-at-2spi.com
Look for us! ############################ WWW: www.2spi.com ############################ ==================================================
Paula Sounds like you have the wrong bulb. An enlarger bulb is opal (milky-white) so the filament should not be visible, and the label should be printed on the base close to the cap, where it won't interfere with image formation . Chris
Dr. Chris Jeffree University of Edinburgh
----- Original Message ----- } From: "Paula Sicurello" {patpxs-at-gwumc.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, August 27, 2003 8:21 PM
Hi,
I am currently testing our new microscopy based reader on which I would like to test a variety of samples. One of the samples I am looking for is a plate which contains 6144 samples of cells or a tissue array within the boundaries of a SBS-standard plate. Arrays or plates with 1536 well or samples/spots on them would be useful too, if they fit into the SBS-standard plate boundaries. The arrays should be in a evenly spaced rectangular pattern. The carrier should be either glass or plastic.
Does anyone have such samples available which I could use for testing either in brightfield mode or with fluorescence or a combination of both ? Preferably fixed material which can be reused several times for testing.
The list of Local Arrangements Committee members shown in the EXPO and Proceedings for the 2003 Microscopy & Microanalysis meeting was based on a preliminary list I submitted two months before the meeting for those publications' deadlines. By the time of the meeting some of the people on the list were unable to participate, and others not on the list attended M&M 2003 and served on the LAC. To give the LAC members proper recognition for the tremendous job they did, here are their names:
} From Texas:
Pamela Neill
Jodi Roepsch
Robert Champaign
Becky Holdford
Ann Burke
Josephine Taylor
Ann Rushing
Susan Robbins
Sandra Westmoreland
Robert Droleskey
} From Oklahoma:
Mary Sheldon
Elizabeth Duncan
Rae Harrington
Amy Sheldon
} From Minnesota:
Dwight Erickson, LAC Treasurer
Thanks to all!
Ev Osten
Local Arrangements Committee Chair Microscopy & Microanalysis 2003 August 3 - 7, San Antonio
efosten-at-mmm.com 651-736-0104 fax: 651-733-0648
3M Company Corporate Analytical Technology Center 3M Center, 201-BE-16 St. Paul, MN 55144-1000
It sounds like you have the point-source bulb in there. The correct bulb for that Durst is a large frosted bulb, the number of which I can no longer remember. If your enlarger is set up for point source, you will need to do a point-source alignment of the system and use the proper condensers to get rid of the filament image. I no longer remember how to do that, either, except that the lens must be used at its largest f/stop. The point source lamp plugs into a rheostat to control the brightness, instead of using regular f-stops, so if you switch to regular condenser lighting, you will need to plug the lamp into a regular outlet. If you later switch back to point-source you MUST remember to switch back to the proper power source or the bulb will explode like a little grenade and scare the *^%*%* out of you. Speaking from experience here. :-)
Lots of luck, Randy
Randy Tindall EM Specialist Electron Microscopy Core---We do Small Well! W122 Veterinary Medicine University of Missouri Columbia, MO 65211 Tel: (573) 882-8304 Fax: (573) 884-5414 Email: tindallr-at-missouri.edu Web: http://www.biotech.missouri.edu/emc/
-----Original Message----- } From: Paula Sicurello [mailto:patpxs-at-gwumc.edu] Sent: Wednesday, August 27, 2003 2:21 PM To: microscopy-at-sparc5.microscopy.com
Hi Listers,
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Barb, I think that I have everything that you would need. Please reply with what you need in particular. I am currently using the Philips 200 and have complete sets of the manuals and I think all the O-rings that you could possible need too. Pat Connelly Research Specialist Dept. of Biology Univ. of Pennsylvania Philadelphia, PA 19104-6018 215-898-7145
Yet another reason to go with Macintosh! Not a problem on our end..
Carol Jean Hirt, VP Materials Research Laboratories, Inc. 800-424-1776 www.mrllab.com
on 8/27/03 12:22 PM, Warren E Straszheim at wesaia-at-iastate.edu wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } This is way off the microscopy topic, but unfortunately, it is relevant to } our daily activities of late. } I am trying to contact someone probably associated with this list to inform } them that they are infected with the SoBig virus and don't appear to know } it yet. } } Like many of us, I have been receiving dozens of copies of the SoBig virus } of late. Being a curious type and being the IT guy for the computers in our } labs, I have been poking into some of the messages to see where they came } from. Even though the Return Path: and From: fields are usually spoofed, I } found that many were coming from a couple of sources. I have included the } relevant information below. } } Since many of the alleged senders were frequent posters to this list I am } guessing that the real sender may be a member of this list. So if BILLPC on } SYMET.NET or PRIVAT on T-DIALIN.NET means something to one of you out } there, please see about getting your PC disinfected. } } Thanks. } } Warren } } Received: from PRIVAT (pD9E7724F.dip.t-dialin.net [217.231.114.79]) } by despam-3.iastate.edu (8.12.4/8.12.4) with ESMTP id h7QCNLKG007344 } for {wesaia-at-iastate.edu} ; Tue, 26 Aug 2003 07:23:22 -0500 } } From: {martin.voecks-at-web.de} } } } Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) } by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7REWjbX018529 } for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 09:32:46 -0500 } Message-Id: {200308271432.h7REWjbX018529-at-despam-2.iastate.edu} } } From: {gary-at-gaugler.com} } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 46 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } } } }
You may wish to consider an alternative. The use of glass slides coated with 2% aminopropyltriethoxysilane in dry acetone is now commonly used to make sections adhere better. You dip once in 2% APTS for 10-30 sec, rinse in acetone and then rinse in distilled water. It works far better than poly-L-lysine or chrome-gel for this purpose. But it was originally described by Gottlieb & Glaser (1976 Biochem Biophys Res Comm 63:815-821) in conjunction with glutaraldehyde to get single cells to adhere to glass better. You might want to check this paper out. good luck. tom
At 09:20 AM 8/27/2003 +0100, you wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
That is one cofiguration and should work OK. I have also used 240/200 and a 105 mm lens. If all you are getting is the filament image, then you need to adjust the bellows and refocus on the grains in your film. From the bulb description, it sounds like you are using a regular enlarging bulb and not a point source. You can verify this by checking if the enlarger is plugged directly into the timer or is there a variable reostat inbetween that allows you to adjust the light intensity. If you need further help in aligment, email me directly.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center Houston TX msteglic-at-mdanderson.org ---------------------- Forwarded by Mannie Steglich/MDACC on 08/28/2003 11:16 AM ---------------------------
Microscopy-request-at-sparc5.microscopy.com on 08/27/2003 02:21:13 PM
To: microscopy-at-sparc5.microscopy.com cc:
Hi Listers,
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Or if you can't live without Windows - just get rid of outlook express on every windows machine you have. Replace it with something better like Eudora that doesn't have so many security flaws.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } Carol Jean Hirt, VP } Materials Research Laboratories, Inc. } 800-424-1776 } www.mrllab.com } } on 8/27/03 12:22 PM, Warren E Straszheim at wesaia-at-iastate.edu wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } This is way off the microscopy topic, but unfortunately, it is relevant to } } our daily activities of late. } } I am trying to contact someone probably associated with this list to inform } } them that they are infected with the SoBig virus and don't appear to know } } it yet. } } } } Like many of us, I have been receiving dozens of copies of the SoBig virus } } of late. Being a curious type and being the IT guy for the computers in our } } labs, I have been poking into some of the messages to see where they came } } from. Even though the Return Path: and From: fields are usually spoofed, I } } found that many were coming from a couple of sources. I have included the } } relevant information below. } } } } Since many of the alleged senders were frequent posters to this list I am } } guessing that the real sender may be a member of this list. So if BILLPC on } } SYMET.NET or PRIVAT on T-DIALIN.NET means something to one of you out } } there, please see about getting your PC disinfected. } } } } Thanks. } } } } Warren } } } } Received: from PRIVAT (pD9E7724F.dip.t-dialin.net [217.231.114.79]) } } by despam-3.iastate.edu (8.12.4/8.12.4) with ESMTP id h7QCNLKG007344 } } for {wesaia-at-iastate.edu} ; Tue, 26 Aug 2003 07:23:22 -0500 } } } From: {martin.voecks-at-web.de} } } } } } } Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) } } by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7REWjbX018529 } } for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 09:32:46 -0500 } } Message-Id: {200308271432.h7REWjbX018529-at-despam-2.iastate.edu} } } } From: {gary-at-gaugler.com} } } } } ------------------------------------------- } } No files should be attached to this message } } ------------------------------------------- } } Warren E. Straszheim, Ph.D. } } Materials Analysis and Research Lab } } Iowa State University } } 46 Town Engineering } } Ames IA, 50011-3232 } } } } Ph: 515-294-8187 } } FAX: 515-294-4563 } } } } E-Mail: wesaia-at-iastate.edu } } Web: www.marl.iastate.edu } } } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } } Computer applications and networking } } } } } } } } } }
Going with Linux or Mac OS will probably ensure that you don't get infected by a virus, but it won't spare you from receiving all the fool traffic that these viruses have generated. These viruses are equal opportunity pains in the butt in that regard.
But I agree with you, Windows is a high-profile target for virus writers. Those of us using it cannot afford to let our guard down and say "it won't happen to me". We should all have safeguards in place and make sure that virus definitions are up to date if we are going to connect to the net.
Then there is also the often forgotten caution, don't open unexpected attachments. I don't care how cute the clip or website is alleged to be, I don't care to risk my computer over that. And if someone is sending me a document, I want to have a clear indication that it is something worthwhile and and authentic and safe. Messages supposedly from colleagues that say simply "Here is the new file - check it out" don't do much to certify that it is a file that I should trust.
BTW, Gordon, one of the virus copies was _allegedly_ from you.
You all be careful out there. Warren
At 12:20 PM 8/28/2003 -0700, you wrote:
} Or better yet, go with Linux... } } Or if you can't live without Windows - just get rid of outlook express on } every windows machine you have. Replace it with something better like } Eudora that doesn't have so many security flaws. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } Carol Jean Hirt, VP } } Materials Research Laboratories, Inc. } } 800-424-1776 } } www.mrllab.com
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Paula, The Durst 45-S should have the two condensor lens configurations printed out on the cover, front panel, of the enlarger head. Some will be for 4x5, 120, and 35mm. Unfortunately I never did much work for 35mm since we had an easy to use Omega with the frosted bulb. A point-source bulb, which is what you have, will give you better images but may also exacerbate problems you may have on the film, such as dust and scratches, which is why many people opt for the frosted set-up, especially for 35mm. Good luck John
-- John Perrino Cell Sciences Imaging Facility Beckman B001 Stanford University Stanford, CA 94305 650-723-3462
Chris, It's not true, halogen bulbs in relatively modern enlargers are transparent. Halogen bulbs delivered much more "light" and it's spectral composition is better for color photography, which, actually, unimportant for B&W. The important thing is that those bulbs considered as a "pointed light source", so the image is sharper and more contrasty.
In my old Devere enlarger, I am using the following combinations:
Condenser: top lens (mm) 180 180 120 bottom 180 120 100 Objective lens------------} 150-105 105-75 60-50 mm
The bulb is transparent, but I am not quite sure it's halogen.
Unfortunately, I know nothing about Durst enlargers.
Sergey
At 12:01 AM 8/28/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Virus from me...? I doubt it. Send me the full headers and I'll check it out. My email is run through pine on a SunOS computer.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Thu, 28 Aug 2003, Warren E Straszheim wrote:
} Going with Linux or Mac OS will probably ensure that you don't get infected } by a virus, but it won't spare you from receiving all the fool traffic that } these viruses have generated. These viruses are equal opportunity pains in } the butt in that regard. } } But I agree with you, Windows is a high-profile target for virus writers. } Those of us using it cannot afford to let our guard down and say "it won't } happen to me". We should all have safeguards in place and make sure that } virus definitions are up to date if we are going to connect to the net. } } Then there is also the often forgotten caution, don't open unexpected } attachments. I don't care how cute the clip or website is alleged to be, I } don't care to risk my computer over that. And if someone is sending me a } document, I want to have a clear indication that it is something worthwhile } and and authentic and safe. Messages supposedly from colleagues that say } simply "Here is the new file - check it out" don't do much to certify that } it is a file that I should trust. } } BTW, Gordon, one of the virus copies was _allegedly_ from you. } } You all be careful out there. } Warren } } At 12:20 PM 8/28/2003 -0700, you wrote: } } } Or better yet, go with Linux... } } } } Or if you can't live without Windows - just get rid of outlook express on } } every windows machine you have. Replace it with something better like } } Eudora that doesn't have so many security flaws. } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } } Gordon Ante Vrdoljak Electron Microscope Lab } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } phone (510) 642-2085 Berkeley CA 94720-3330 } } fax (510) 643-6207 cell (510) 290-6793 } } } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } } } Carol Jean Hirt, VP } } } Materials Research Laboratories, Inc. } } } 800-424-1776 } } } www.mrllab.com } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 46 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } }
I've been asked to see what the EM difference is between rat eye lens epithelium in culture and in situ. So I'd like to fix the epithelium while it is still in the eye. I'm worried that if I do that it then won't be possible to get the epithelium off the lens without damaging it; I also seem to remember that lens becomes brittle and difficult to handle when fixed. I'd like to keep life simple, so it may be necessary to take the epithelium off the lens before fixing. Does anyone have experience with this?
Perhaps this is a good time to engage PDF as safe attachments(?). It would seem so.
The Mac is not particularly affected since it constitutes a small percentage of PC sales and total installed base. However, it is not immune. It is just that nobody wastes time on a minuscule platform. If it is hyped-up, of course that may change.
gary g.
At 01:30 PM 8/28/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The main thing with fixing eyes is to make sure that the fix can get in - so slit across the cornea and take out the lens (assuming you don't want either of these). For EM, glut is OK, but because an eye is big, the fast penetration/fixation of paraform combined with the thorough action of glut is better - so Karnovsky's. If you need to do immuno, neither of these is suitable; PLP (paraform/lysine/periodate) is good or just paraform. As for McD-Trump - I've never heard of it!
Diana
At 12:19 PM -0400 26/8/03, Louro, Pedro wrote: } I'm looking for any feedback on fixation of eyes (small and large animal). } More specifically: a fixative that would work well for Histology and EM. } Any info. on: } * Glutaraldehyde } * Karnovsky's Fixative } * McDowell - Trump's Fixative } or any other fixative would be greatly appreciated. } } Thanks in advance, } } Pedro
--
Diana van Driel Department of Clinical Ophthalmology Sydney University GPO Box 4337 Sydney NSW AUSTRALIA 2001
My linux server has not noticed, is there a new virus ?
but my windows box cannot find time to shut out the viri, the popups, the the cookies and the adware, spyware, and other stuff. Could it be because some of the browsers were written to accomodate, to force the user to put up with these things? Browsers simply read the file that are transmitted from the server to the client. If the browsers ignored the adware, the pop ups and stuff, there would be few people sending them out, because no one could read them.
What is needed is concerted organized effort by those of us in the Internet community to find the people that entered this virus into the system.
Interesting thought, back in 1947, 48, and 49 there was a similar problem with people on the radio, everybody kept trying to out maneuver the next guy so that no one could get anything on their personal radio receivers.
It turned out that the big stations were heavily financed and they wanted the little guys (competition) off the air. So they thought, we will keep doing this until we can get the government to license the airways, in that manner we can control the media called radio by forcing them to get licensed, and by limiting the number of licenses, and eliminate our competition, since the little guys do not have money, manpower, or knowledge about how to lobby the government.
Now this is not a result we want to happen with the internet. Government should not be involved in the email system. So let's start finding the persons that did this So Big Virus. Lets enlist everyone in the net to help back track it. until we find its source. That should not be too big of a job. it only happened last week.
I am betting the guys responsible for it, would suprise us all?
Any ideas how we could back trace this? one idea is two create two threads on this list: 1. to identify So Big by its entry date and source amoung members 2. the other to report any progress in backtracking to the origin that is known or reported by anyone anywhere through out the world as one or more of us becomes aware of it.
sterling
On Thu, 28 Aug 2003, Warren E Straszheim wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Going with Linux or Mac OS will probably ensure that you don't get infected } by a virus, but it won't spare you from receiving all the fool traffic that } these viruses have generated. These viruses are equal opportunity pains in } the butt in that regard. } } But I agree with you, Windows is a high-profile target for virus writers. } Those of us using it cannot afford to let our guard down and say "it won't } happen to me". We should all have safeguards in place and make sure that } virus definitions are up to date if we are going to connect to the net. } } Then there is also the often forgotten caution, don't open unexpected } attachments. I don't care how cute the clip or website is alleged to be, I } don't care to risk my computer over that. And if someone is sending me a } document, I want to have a clear indication that it is something worthwhile } and and authentic and safe. Messages supposedly from colleagues that say } simply "Here is the new file - check it out" don't do much to certify that } it is a file that I should trust. } } BTW, Gordon, one of the virus copies was _allegedly_ from you. } } You all be careful out there. } Warren } } At 12:20 PM 8/28/2003 -0700, you wrote: } } } Or better yet, go with Linux... } } } } Or if you can't live without Windows - just get rid of outlook express on } } every windows machine you have. Replace it with something better like } } Eudora that doesn't have so many security flaws. } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } } Gordon Ante Vrdoljak Electron Microscope Lab } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } phone (510) 642-2085 Berkeley CA 94720-3330 } } fax (510) 643-6207 cell (510) 290-6793 } } } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } } } Carol Jean Hirt, VP } } } Materials Research Laboratories, Inc. } } } 800-424-1776 } } } www.mrllab.com } } ------------------------------------------- } No files should be attached to this message } ------------------------------------------- } Warren E. Straszheim, Ph.D. } Materials Analysis and Research Lab } Iowa State University } 46 Town Engineering } Ames IA, 50011-3232 } } Ph: 515-294-8187 } FAX: 515-294-4563 } } E-Mail: wesaia-at-iastate.edu } Web: www.marl.iastate.edu } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } Computer applications and networking } } }
Does anyone know what has become of DiaTech in Knoxville Tenn.? They were makers of fine diamond knives for ultramicrotomy and must have folded or been sold on to another company--any news of them? Many thanks, Lee F. Brunckhorst leeb-at-uow.edu.au (Wollongong University Materials Engineering, Australia)
I'll have to go with Gary on this one, at the risk of starting a flame war. My Timex Sinclair and Atari ST are completely immune to viruses for the same reason...
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Hi, Could those of you doing EBSD in an SEM kindly share your sample prep method with me. I welcome all feedback, but would particularly like to here from those working with geological samples. Is anyone using an Ion Mill? If so, what is the maximum size of the 'thinned area'?
TIA Mike -- ******************************************************************** Michael M. Cheatham 312 Heroy Geology Laboratory Phone (315)-443-1261 Syracuse University Fax (315)-443-3363 Syracuse, NY 13244-1070
owner of PLASMACHEM-L: http://listserv.syr.edu/archives/plasmachem-l.html owner of XRF-L: http://listserv.syr.edu/archives/xrf-l.html owner of TIMS-L: http://listserv.syr.edu/archives/tims-l.html owner of SIRIS-L: http://listserv.syr.edu/archives/siris-l.html ********************************************************************
Mannie is correct in suggesting to adjust the bellow to focus the grain on your negative. Specifically, you should bring the 50mm lens closer to the condensers. You were at a wrong focal plane when you see filament. It does not matter what bulb you use in terms of focusing.
Ann Fook Yang
} } } {"msteglic-at-mdanderson.org"-at-sparc5.microscopy.com} 08/28/03 12:22PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
That is one cofiguration and should work OK. I have also used 240/200 and a 105 mm lens. If all you are getting is the filament image, then you need to adjust the bellows and refocus on the grains in your film. From the bulb description, it sounds like you are using a regular enlarging bulb and not a point source. You can verify this by checking if the enlarger is plugged directly into the timer or is there a variable reostat inbetween that allows you to adjust the light intensity. If you need further help in aligment, email me directly.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center Houston TX msteglic-at-mdanderson.org ---------------------- Forwarded by Mannie Steglich/MDACC on 08/28/2003 11:16 AM ---------------------------
Microscopy-request-at-sparc5.microscopy.com on 08/27/2003 02:21:13 PM
To: microscopy-at-sparc5.microscopy.com cc:
Hi Listers,
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
Mannie is correct in suggesting to adjust the bellow to focus the grain on your negative. Specifically, you should bring the 50mm lens closer to the condensers. You were at a wrong focal plane when you see filament. It does not matter what bulb you use in terms of focusing.
Ann Fook Yang
} } } {"msteglic-at-mdanderson.org"-at-sparc5.microscopy.com} 08/28/03 12:22PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
That is one cofiguration and should work OK. I have also used 240/200 and a 105 mm lens. If all you are getting is the filament image, then you need to adjust the bellows and refocus on the grains in your film. From the bulb description, it sounds like you are using a regular enlarging bulb and not a point source. You can verify this by checking if the enlarger is plugged directly into the timer or is there a variable reostat inbetween that allows you to adjust the light intensity. If you need further help in aligment, email me directly.
Mannie Steglich Tech Dir E M Lab U T M D Anderson Cancer Center Houston TX msteglic-at-mdanderson.org ---------------------- Forwarded by Mannie Steglich/MDACC on 08/28/2003 11:16 AM ---------------------------
Microscopy-request-at-sparc5.microscopy.com on 08/27/2003 02:21:13 PM
To: microscopy-at-sparc5.microscopy.com cc:
Hi Listers,
I have a Durst Laborator S-45 and want to do some 35mm work. I have the proper big glass lenses (130/85), the proper small lense (50mm).
I haven't do 35mm in a long time so I can't remember the complete setup. When I put in the 50mm lens in the turret and put the 130/85 in the proper configuration all I get is a lovely image of the filament of the light bulb that is the light source.
Does anybody know how to set this up right? I thought when I did this a looooong time ago that all I did was swap the big glass lenses and away I went.
Could it be that I don't have the correct light source in there? It's a smallish regular looking (by that I mean it looks like a bulb you use at home) clear glass light bulb.
Any suggestions are greatly appreciated. Of course I don't have/can't find the instruction manual that would to be convenient.
Dodging summer thunderstorms,
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Electron Microscope Lab Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
I am glad you mentioned PDF, i have noticed that when I load some PDf's that all of a sudden my computer starts sending things outbound lots and lots of bits and I cannot stop it unless I shut down and reboot. anyone else notice that with PDF? Can someone say what it is that is happening?
sterling
On Thu, 28 Aug 2003, Gary Gaugler wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Perhaps this is a good time to engage PDF } as safe attachments(?). It would seem so. } } The Mac is not particularly affected since it } constitutes a small percentage of PC sales } and total installed base. However, it is not immune. } It is just that nobody wastes time on a minuscule } platform. If it is hyped-up, of course that } may change. } } gary g. } } } At 01:30 PM 8/28/2003, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Going with Linux or Mac OS will probably ensure that you don't get } } infected by a virus, but it won't spare you from receiving all the fool } } traffic that these viruses have generated. These viruses are equal } } opportunity pains in the butt in that regard. } } } } But I agree with you, Windows is a high-profile target for virus writers. } } Those of us using it cannot afford to let our guard down and say "it won't } } happen to me". We should all have safeguards in place and make sure that } } virus definitions are up to date if we are going to connect to the net. } } } } Then there is also the often forgotten caution, don't open unexpected } } attachments. I don't care how cute the clip or website is alleged to be, I } } don't care to risk my computer over that. And if someone is sending me a } } document, I want to have a clear indication that it is something } } worthwhile and and authentic and safe. Messages supposedly from colleagues } } that say simply "Here is the new file - check it out" don't do much to } } certify that it is a file that I should trust. } } } } BTW, Gordon, one of the virus copies was _allegedly_ from you. } } } } You all be careful out there. } } Warren } } } } At 12:20 PM 8/28/2003 -0700, you wrote: } } } } } Or better yet, go with Linux... } } } } } } Or if you can't live without Windows - just get rid of outlook express on } } } every windows machine you have. Replace it with something better like } } } Eudora that doesn't have so many security flaws. } } } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } } } Gordon Ante Vrdoljak Electron Microscope Lab } } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } } phone (510) 642-2085 Berkeley CA 94720-3330 } } } fax (510) 643-6207 cell (510) 290-6793 } } } } } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } } } } } Carol Jean Hirt, VP } } } } Materials Research Laboratories, Inc. } } } } 800-424-1776 } } } } www.mrllab.com } } } } ------------------------------------------- } } No files should be attached to this message } } ------------------------------------------- } } Warren E. Straszheim, Ph.D. } } Materials Analysis and Research Lab } } Iowa State University } } 46 Town Engineering } } Ames IA, 50011-3232 } } } } Ph: 515-294-8187 } } FAX: 515-294-4563 } } } } E-Mail: wesaia-at-iastate.edu } } Web: www.marl.iastate.edu } } } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } } Computer applications and networking } } } }
} Perhaps this is a good time to engage PDF } as safe attachments(?). It would seem so. } } The Mac is not particularly affected since it } constitutes a small percentage of PC sales } and total installed base. However, it is not immune. } It is just that nobody wastes time on a minuscule } platform. If it is hyped-up, of course that } may change. } } gary g.
Actually also not a good idea, there are such things as a PDF virus.
The biggest problem with Windows (any flavor) is Outlook Express. As installed, it auto executes any Attachment. This makes it a sucker punch for the common cracker. If Windows users turned that "feature" off or use a different email app (like Eudora) then virus propagation would drop by many factors. This is how the SoBig virus infects and propagates.
Another problem is Windows (any flavor), by default, has many network ports open that have no business being open thus exposing the machine to numerous buffer overflow cracks. This is how the MSBlaster virus infects and propagates.
As for the MacOS not being affected because the size of it's market. A small gain of truth but mostly FUD. Market share of linux is also small but that does not keep the virus and cracks away. Also statistically, even though the MacOS is smaller, one should see a few virus problems over the years. What do I see, zero, nothing. I don't even run virus protection software on the MacOS any more. Stopped doing that many years ago.
As a professional programmer fluent in apps and drivers in both the WinOS and MacOS world, the truth is there are fundamental differences in the way OS9 works from Windows that make 95% of the virus cracks impossible to implement. Can you design a virus to infect MacOS9, yes but you are going to have to work really hard at it. Script kiddies and hack virus programmers need not apply.
OSX is also different but OSX is an industrial strength OS that is installed in a safe manner (for example, there are no network ports open unless you open them. Note that does not mean you can't surf the net, it means that no one can surf you). Also Apple takes a defensive approach, they assume that there are evil people out there that want to mess with your computer. Microsoft does not design Windows with security in mind, they keep saying they are but as long as those extraneous network ports are open by default, I know they are not.
As a case in point, 4pi runs an powerpc linux server for web, ftp and mail services. We always get probed and I mean seriously probed, but they can't get in because 1) it's a powerpc box and intel linux cracks don't work. and 2) it's locked down with regard to security. We have all flavors of OSs here. Macs (both OS9 and OSX) and linux boxes are permitted email access. Windows boxes are not. Too much risk with cracks. Last time I saw a virus problem on a MacOS box was back in the pre-powerpc days and that was an programming experiment in self infection.
For more years, than I can count, non-MacOS users have been declaring victory and Apple with the MacOS dead because of its size. Apple is still around, Macs are still around and with OSX, the market size is even growing. OSX is pretty neat I must say.
And last, you will not keep out a professional cracker out of your computer in the same way you will not keep a professional thief out of your house. Professional are a different breed, they will get in. That's way they are called professionals. Thankfully, most all virus and crack writers are not real professionals. I can recommend several books that will put the fear of god in anyone that thinks the internet is a benign place. It's a real war zone and the sooner we accept that fact and convince companies such as Microsoft to default the OS in a secure mode, then we will start to see a reduction rather than a increase in cracks.
Microsoft resorted to linux to protect its servers. I found it funny. http://www.internetweek.com/story/showArticle.jhtml?articleID=13100775
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ Gordon Ante Vrdoljak Electron Microscope Lab ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall gvrdolja-at-nature.berkeley.edu UC Berkeley phone (510) 642-2085 Berkeley CA 94720-3330 fax (510) 643-6207 cell (510) 290-6793
On Thu, 28 Aug 2003, Gordon Vrololjak wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Or better yet, go with Linux... } } Or if you can't live without Windows - just get rid of outlook express on } every windows machine you have. Replace it with something better like } Eudora that doesn't have so many security flaws. } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } Gordon Ante Vrdoljak Electron Microscope Lab } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } gvrdolja-at-nature.berkeley.edu UC Berkeley } phone (510) 642-2085 Berkeley CA 94720-3330 } fax (510) 643-6207 cell (510) 290-6793 } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } Carol Jean Hirt, VP } } Materials Research Laboratories, Inc. } } 800-424-1776 } } www.mrllab.com } } } } on 8/27/03 12:22 PM, Warren E Straszheim at wesaia-at-iastate.edu wrote: } } } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } To Subscribe/Unsubscribe -- } } } http://www.msa.microscopy.com/MicroscopyListserver } } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } } } } This is way off the microscopy topic, but unfortunately, it is relevant to } } } our daily activities of late. } } } I am trying to contact someone probably associated with this list to inform } } } them that they are infected with the SoBig virus and don't appear to know } } } it yet. } } } } } } Like many of us, I have been receiving dozens of copies of the SoBig virus } } } of late. Being a curious type and being the IT guy for the computers in our } } } labs, I have been poking into some of the messages to see where they came } } } from. Even though the Return Path: and From: fields are usually spoofed, I } } } found that many were coming from a couple of sources. I have included the } } } relevant information below. } } } } } } Since many of the alleged senders were frequent posters to this list I am } } } guessing that the real sender may be a member of this list. So if BILLPC on } } } SYMET.NET or PRIVAT on T-DIALIN.NET means something to one of you out } } } there, please see about getting your PC disinfected. } } } } } } Thanks. } } } } } } Warren } } } } } } Received: from PRIVAT (pD9E7724F.dip.t-dialin.net [217.231.114.79]) } } } by despam-3.iastate.edu (8.12.4/8.12.4) with ESMTP id h7QCNLKG007344 } } } for {wesaia-at-iastate.edu} ; Tue, 26 Aug 2003 07:23:22 -0500 } } } } From: {martin.voecks-at-web.de} } } } } } } } } } Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) } } } by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7REWjbX018529 } } } for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 09:32:46 -0500 } } } Message-Id: {200308271432.h7REWjbX018529-at-despam-2.iastate.edu} } } } } From: {gary-at-gaugler.com} } } } } } } ------------------------------------------- } } } No files should be attached to this message } } } ------------------------------------------- } } } Warren E. Straszheim, Ph.D. } } } Materials Analysis and Research Lab } } } Iowa State University } } } 46 Town Engineering } } } Ames IA, 50011-3232 } } } } } } Ph: 515-294-8187 } } } FAX: 515-294-4563 } } } } } } E-Mail: wesaia-at-iastate.edu } } } Web: www.marl.iastate.edu } } } } } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } } } Computer applications and networking } } } } } } } } } } } } } } } } }
What you describe sounds like you are mounting the 50mm lens below the lens mounting turret in the same manner as you mount the 135mm and 150mm lenses for larger format negatives.
The 50mm lens must be recessed into the bellows. There is a tube shaped mounting adapter that fits into the lens mounting turret.
Frank **************************************************************************** Frank Macaluso tel: 718-430-3547 Analytical Imaging Facility fax: 718-430-8996 Albert Einstein College of Medicine e-mail: macaluso-at-aecom.yu.edu 1300 Morris Park Avenue http://www.aecom.yu.edu/aif/ Bronx, NY 10461 ****************************************************************************
No, the virus was not from you. I said "_allegedly_" from you. You can check out the headers below. The culprit was BILLPC, whoever that may be. Your name (and many other listers including myself and Debby Sherman, etc) have been forged as the sources. I just wanted you to know you were included in the party. {G}
More viruses are hiding or forging the identity of the true sender. I don't assume anymore that the alleged source is the real source but instead examine the other headers before I try to alert someone to an infection on their end. I have been scolded as the source of a virus on several such occasions. In all cases, so far, I have not been the real source.
Warren
At 03:05 PM 8/28/2003 -0700, Gordon V. wrote: } Virus from me...? } I doubt it. Send me the full headers and I'll check it out. My email is } run through pine on a SunOS computer.
Received: from BILLPC (dsl1-216-90-9-53.symet.net [216.90.9.53]) by despam-2.iastate.edu (8.12.4/8.12.4) with ESMTP id h7RKJFbX022026 for {wesaia-at-iastate.edu} ; Wed, 27 Aug 2003 15:19:17 -0500 Message-Id: {200308272019.h7RKJFbX022026-at-despam-2.iastate.edu} } From: {gvrdolja-at-nature.berkeley.edu} To: {wesaia-at-iastate.edu}
I suppose it could be one of a couple things with PDF files.
First, I think PDF files may now allow for some code, either macros or embedded files, that could execute on your computer when you view the file. I would need a real PDF expert to comment on the current capabilities. I know MS Word document files were immune from infection until Word 97, IIRC. About that time, MS started supporting macros as valid parts of Word documents. It wasn't long before hackers started exploiting the "feature". Even before that, the Concept virus came out using a document template (.DOT) named as a document (.DOC) showing that documents could now be hosts for viruses. Maybe the same thing is happening to PDF files.
Second, it could be that PDF is not the real file extension. I know I have seen attachments with names such as "details.txt.vbs". The file could show up as "details.txt" in the mail window or an Explorer window if the system was set to hide known file extensions. Unfortunately, the default Windows installation hides extensions, ala Mac, and allows hackers to use this trick to fool users into thinking the file is just a benign text or PDF file. But when the file is executed, Windows uses the true, hidden extension to determine what to do with the file.
Either way, I come back to the conclusion that attachments should be received only cautiously. I appreciate Scott Davilla's comments that the Internet is a very dangerous place. If we knew just how dangerous, we might be more careful.
Warren
At 02:23 AM 8/29/2003 -0500, you wrote:
} I am glad you mentioned PDF, i have noticed that when I load some PDf's } that all of a sudden my computer starts sending things outbound lots and } lots of bits and I cannot stop it unless I shut down and reboot. anyone } else notice that with PDF? } Can someone say what it is that is happening? } } sterling
------------------------------------------- No files should be attached to this message ------------------------------------------- Warren E. Straszheim, Ph.D. Materials Analysis and Research Lab Iowa State University 46 Town Engineering Ames IA, 50011-3232
Hello all, I am looking into getting a nebulizer to deposit proteins in glycerol as 25-150um diameter droplets onto cleaved sheets of mica that will later be rotary shadowed. I have heard that the bulb-driven nebulizers do not give enough condensed force of spray to create fine droplets. Does anyone have experience with this method and can suggest protocols and a nebulizer to purchase? John -- John Perrino Cell Sciences Imaging Facility Beckman B001 Stanford University Stanford, CA 94305 650-723-3462
Can anyone recommend a source for an antibody against rabbit-GFP to use for TEM immunocytochemistry? We are trying to localize a protein with a GFP construct in arabidopsis root tissue. We have tried one antibody but labeling is poor and not well localized. We want to make sure that it is not the antibody. The protein is an unknown and the research group has not yet been able to develop a clean antibody against it.
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
Apparently they may have found him http://www.reuters.com/newsArticle.jhtml;jsessionid=1MBT2YBXGTM1QCRBAELCFEY?type=topNews&storyID=3361073
On Thu, 28 Aug 2003 sstouden-at-thelinks.com wrote:
} } My linux server has not noticed, is there a new virus ? } } but my windows box cannot find time to shut out the viri, the popups, the } the cookies and the adware, spyware, and other stuff. Could it be because } some of the browsers were written to accomodate, to force the user to put } up with these things? Browsers simply read the file that are transmitted } from the server to the client. If the browsers ignored the adware, the } pop ups and stuff, there would be few people sending them out, because no } one could read them. } } What is needed is concerted organized effort by those of us in the } Internet community to find the people that entered this virus into the } system. } } Interesting thought, back in 1947, 48, and 49 there was a similar problem } with people on the radio, everybody kept trying to out maneuver the next } guy so that no one could get anything on their personal radio receivers. } } It turned out that the big stations were heavily financed and they wanted } the little guys (competition) off the air. So they thought, we will keep } doing this until we can get the government to license the airways, in } that manner we can control the media called radio by forcing them to } get licensed, and by limiting the number of licenses, and eliminate our } competition, since the little guys do not have money, manpower, or } knowledge about how to lobby the government. } } Now this is not a result we want to happen with the internet. Government } should not be involved in the email system. So let's start finding the } persons that did this So Big Virus. Lets enlist everyone in the net to } help back track it. until we find its source. That should not be too big } of a job. it only happened last week. } } } I am betting the guys responsible for it, would suprise us all? } } Any ideas how we could back trace this? } one idea is two create two threads on this list: } 1. to identify So Big by its entry date and source amoung members } 2. the other to report any progress in backtracking to the origin } that is known or reported by anyone anywhere through out the } world as one or more of us becomes aware of it. } } sterling } } } On Thu, 28 Aug 2003, Warren E Straszheim wrote: } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Going with Linux or Mac OS will probably ensure that you don't get infected } } by a virus, but it won't spare you from receiving all the fool traffic that } } these viruses have generated. These viruses are equal opportunity pains in } } the butt in that regard. } } } } But I agree with you, Windows is a high-profile target for virus writers. } } Those of us using it cannot afford to let our guard down and say "it won't } } happen to me". We should all have safeguards in place and make sure that } } virus definitions are up to date if we are going to connect to the net. } } } } Then there is also the often forgotten caution, don't open unexpected } } attachments. I don't care how cute the clip or website is alleged to be, I } } don't care to risk my computer over that. And if someone is sending me a } } document, I want to have a clear indication that it is something worthwhile } } and and authentic and safe. Messages supposedly from colleagues that say } } simply "Here is the new file - check it out" don't do much to certify that } } it is a file that I should trust. } } } } BTW, Gordon, one of the virus copies was _allegedly_ from you. } } } } You all be careful out there. } } Warren } } } } At 12:20 PM 8/28/2003 -0700, you wrote: } } } } } Or better yet, go with Linux... } } } } } } Or if you can't live without Windows - just get rid of outlook express on } } } every windows machine you have. Replace it with something better like } } } Eudora that doesn't have so many security flaws. } } } } } } \/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ } } } Gordon Ante Vrdoljak Electron Microscope Lab } } } ICQ 23243541 http://nature.berkeley.edu/~gvrdolja 26 Giannini Hall } } } gvrdolja-at-nature.berkeley.edu UC Berkeley } } } phone (510) 642-2085 Berkeley CA 94720-3330 } } } fax (510) 643-6207 cell (510) 290-6793 } } } } } } On Thu, 28 Aug 2003, Materials Research Laboratories, Inc. wrote: } } } } } } } } Yet another reason to go with Macintosh! Not a problem on our end.. } } } } } } } } Carol Jean Hirt, VP } } } } Materials Research Laboratories, Inc. } } } } 800-424-1776 } } } } www.mrllab.com } } } } ------------------------------------------- } } No files should be attached to this message } } ------------------------------------------- } } Warren E. Straszheim, Ph.D. } } Materials Analysis and Research Lab } } Iowa State University } } 46 Town Engineering } } Ames IA, 50011-3232 } } } } Ph: 515-294-8187 } } FAX: 515-294-4563 } } } } E-Mail: wesaia-at-iastate.edu } } Web: www.marl.iastate.edu } } } } Scanning electron microscopy, x-ray analysis, and image analysis of materials } } Computer applications and networking } } } } } } } }
As the recipient of an excellent grant, I am contemplating a purchase of a digital camera for my High School classroom, to be used often on microscopes of various kinds---with 23mm and 30mm id eyepiece tubes, one of them a trinocular scope. These are not "up-to-date" scopes. Be that as it may. I would like to request comments from list members, who in the past have given lots of such advice. I apologize for covering familiar territory.
I note that many microscopists have mentioned Coolpix's on this list. I am interested in probably either a Coolpix or a Canon 1D EOS digital camera.
Although I would appreciate comments from users of the Canon, I fear that the 1:1.6 cmos photodetector size to 35mm film size would render it difficult to focus the EOS with a straightforward adaptor such as the Diagnostic Instruments adaptors for 35mm cameras---which works QUITE well with a Canon film EOS. THis was my plan, but, again, I am pretty discouraged by potential focusing problems.
I have ruled out the Olympus E20N, after some experience and alot of vignetting.
The Coolpix 5000 looks interesting. It does much of what I need, including monitoring the live specimen on a TV, Computer, or video projector. I like some of the features of the 5700, including better telephoto range for shore bird studies. I have a couple of questions I would refer to those with Coolpix experience:
How well does the 5700 work on a scope? Does the 5700 identically to the 5000 out of the box, with the Nikon adaptor kit? Are these cameras significantly better than the 4500? Are macro kits available to enable focus to within 1/8"? (I intend to use it as an "aquarium microscope" for example to study the community at the sediment/water column interface, for which the Videoflex camera works admirably well.) Can one take a shot easily an within ordinary lag values while monitoring in real time? Nikon has a relay lens (is that a projection lens?) for this purpose, that will work on Coolpix 5000; does it work ok on the 5700?
Another question: is the Nikon D-100 worth the extra cost, and does it incorporate the video monitoring Coolpix features? Is it useable on a scope, and how?
Thank you for any comments or suggestions. I apologize for asking so many questions at once...
Alan Davis Marianas High School Saipan
I wish to ask whether microscopists feel it
-- adavis-at-saipan.com 1-670-322-6580 Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI
I have steadily endeavored to keep my mind free, so as to give up any hypothesis, however much beloved -- and I cannot resist forming one on every subject -- as soon as facts are shown to be opposed to it. -- Charles Darwin (1809-1882)
The right to search for truth implies also a duty; one must not conceal any part of what one has recognized to be true. -- Albert Einstein
As we enjoy great advantages from the inventions of others we should be glad of an opportunity to serve others by any invention of ours, and this we should do freely and generously. -- Benjamin Franklin
Debby- Usually, GFP is fluorescent or luminesces on it own part, and it isn't necessary to localize it with an antibody.
Here, there seems to be some confusion about the subject protein. GFP is not a rabbit protein. If the protein of interest is being localized with a rabbit antibody, that makes sense. But what is the reagent with the GFP? Carol Heckman (Bowling Green State University)
} Date: Fri, 29 Aug 2003 17:28:42 -0500 } Subject: GFP antibody } From: Debby Sherman {dsherman-at-purdue.edu} } To: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} } X-Virus-Scanned: by amavisd-new on Purdue Mailhub } X-Sig: 11b694fff7e42b4e8f56e22d68ad0d62 } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Carol GFP is fluorescent, but there are many uses for antibodies against it nevertheless, including immunoblotting, immunoprecipitation and detection of the GFP-labelled protein by immunogold labelling for electron microscopy in the absence of antibodies against the target protein itself.
What is presumably being requested is a recommendation for a reliable anti-GFP.
Chris
----- Original Message ----- } From: "Carol Heckman" {heckman-at-bgnet.bgsu.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Sunday, August 31, 2003 12:19 AM
Carol,
You are correct in that GFP is often inserted into the code of a particular protein so that it is replicated as the protein in replicated. Then the fluorescent properties of the GFP molecule can be used to localize the particular target protein using fluorescent or confocal microscopy. However, this fluorescence is not useful if you want to do higher resolution localization of the protein. For that you need TEM immunocytochemical procedures utilizing a marker such as colloidal gold.
It is very helpful to identify GFP in living tissue and then take that same tissue, prepare it for TEM ICC and then localize the protein using an antibody against the GFP. If that antibody is made in rabbits than you need an anti-rabbit colloidal gold IgG probe.
What I wanted was suggestions on where to get the antibody against GFP that was been successfully used for TEM ICC. I would like to try a second antibody to make sure my localization, or lack of such, is not due to the antibody we have. Often an antibody will work well in Westerns, such as the one we have, and not work as well under ICC conditions. This is just like how one rabbit will produce what looks like a great antibody and another rabbit will produce one that does not appear to be as strong but actually will work better. Don't ask me why,...it just happens.
I did lead you astray in that I said rabbit -GFP in my original post. I am sorry for that in that It should have read rabbit antibody against GFP. We always have lots of anti-rabbit colloidal gold tagged IgG around so prefer rabbit antibodies. However, we can easily get IgG from other sources so that should not be a limit if there is a GFP antibody out there from another organism that works better than the rabbit ones.
Debby
On 8/30/03 6:19 PM, "Carol Heckman" {heckman-at-bgnet.bgsu.edu} wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Debby- } Usually, GFP is fluorescent or luminesces on it own part, and it } isn't necessary to localize it with an antibody. } } Here, there seems to be some confusion about the subject protein. } GFP is not a rabbit protein. If the protein of interest is being } localized with a rabbit antibody, that makes sense. But what is the } reagent with the GFP? } Carol Heckman } (Bowling Green State University) } } } Date: Fri, 29 Aug 2003 17:28:42 -0500 } } Subject: GFP antibody } } From: Debby Sherman {dsherman-at-purdue.edu} } } To: "message to: MSA list" {microscopy-at-sparc5.microscopy.com} } } X-Virus-Scanned: by amavisd-new on Purdue Mailhub } } X-Sig: 11b694fff7e42b4e8f56e22d68ad0d62 } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- } } http://www.msa.microscopy.com/MicroscopyListserver } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } Hi, } } } } Can anyone recommend a source for an antibody against rabbit-GFP to use } } for TEM immunocytochemistry? We are trying to localize a protein with a GFP } } construct in arabidopsis root tissue. We have tried one antibody but } } labeling is poor and not well localized. We want to make sure that it is } } not the antibody. The protein is an unknown and the research group has not } } yet been able to develop a clean antibody against it. } } } } Debby } } } } Debby Sherman, Manager Phone: 765-494-6666 } } Life Science Microscopy Facility FAX: 765-494-5896 } } Purdue University E-mail: dsherman-at-purdue.edu } } S-052 Whistler Building } } 170 S. University Street } } West Lafayette, IN 47907 } } }
Apologies for the late reply. I don't want to overdo this topic, but few peeps have addressed one of the original questions: is there much danger to getting abrasive sloshed in your eye?
While SiC/water splashes might not be much of a chemical irritant, aqueous suspensions of metal oxide particles are typically not pH-neutral, e.g., alumina suspensions--} acidic, silica suspensions--} basic. So, if you work with those types of polishing suspensions, an eye splash seems likely to be more painful than just the irritation associated with having gritty stuff in your eye.
I work in a lab where I wear safety glasses and frequently go back and forth between the polisher and the optical microscope. It's a bit of a pain to take the glasses off and on, but you get used to it, and the slight inconvenience is worth preventing a splash in the eye.
Eve
-- Eve Donnelly Experimental Biomechanics Lab Sibley School of Mechanical & Aerospace Engineering Cornell University 130 Upson Hall Ithaca, NY 14853 tel. 607.255.3582 fax. 607.255.1222
I have two suggestions: 1. Cryo sections of the same should help 2. Application of alcohol helps to loosen the epithelium which can then be gently peeled off. This techique is used to debride the corneal epithelium during surgeries. Shashi CCMB, Hyderabad INDIA
Hi all,
I've been asked to see what the EM difference is between rat eye lens epithelium in culture and in situ. So I'd like to fix the epithelium while it is still in the eye. I'm worried that if I do that it then won't be possible to get the epithelium off the lens without damaging it; I also seem to remember that lens becomes brittle and difficult to handle when fixed. I'd like to keep life simple, so it may be necessary to take the epithelium off the lens before fixing. Does anyone have experience with this?
Thanks,
Diana
===== Shashi Singh Scientist Centre for Cellular and Molecular Biology Hyderabad-500 007 INDIA PH-91-40-7192575,7192761,7192615 FAX-91-40-7160591, 7160311
__________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com
Temporary position (contractor status) offering 40 hrs/week with limited benefits
Job Description: Transmission Electron Microscopist / Analyst in Process Characterization Lab supporting the Advanced Technology Development Facility (ATDF) as well as both internal and external projects at International SEMATECH (ISMT) in Austin, Texas. The TEM group at ISMT provides materials characterization in support of research and process development projects for future integrated circuit manufacturing. Our research projects tend to focus on materials issues several years ahead of current semiconductor industry manufacturing trends, we therefore often study materials systems that offer new and exciting characterization problems and often collaborate with leading researchers at national labs and universities around the world, however we also have a significant workload involving routine characterization and process monitoring for systems that are already well understood.
Required: M.S. or Ph.D. in Materials Science, Solid-State Physics or Chemistry with strong analytical TEM experience.
For more details please contact: Brendan Foran -at- Brendan_Foran-at-SEMATECH.org
International SEMATECH, headquartered in Austin, Texas, is a global consortium of leading semiconductor manufacturers that represent about half the world's semiconductor production. International SEMATECH engages in cooperative, precompetitive efforts to improve semiconductor manufacturing technology through the support of our members. To learn more about International SEMATECH, visit our website at www.sematech.org.
} not related to the real safety (mostly keeping paperwork in a good shape } and accurately fill out chemical waste disposal tags). So, in such } situation, I think, it's superviser's personal responsibility to provide } safely environment to the workers and train them accordingly.
Look, we don't want to have this devolve into a session on what is safe and what isn't, but this is a very simplistic view. Paperwork may be essential for real safety of all handlers of material down the chain of disposal. If a supervisor has a limited budget, he/she is going to scrimp on safety.
Policies are in place to protect workers who historically have been mistreated. Yes, blanket policies do not apply in all situations. The bureaucracy simply needs some flexibility for individually documented exemptions.
} P.S. By the way: in my EM work, my glasses, I wear to correct my vision, } save my eyes in uncounted number of times. I, also, used to remove them } every time I am using light microscope or binoculars. I don't see big } problem removing them if necessary. But: yes, it decreases my } productivity. Ok, let say, I need about 5 sec. to remove them and put } back. Let say I do it 40 times a day: 5 secx40=200 sec/3.3 min per day.
But you're wearing gloves and your gloves shouldn't be near your unshielded eyes each time you have to handle the googles. So you really should take off your gloves before removing your goggles each time.
____________________________________________________________________________ Michael Cammer Analytical Imaging Facility Albert Einstein Coll. of Med. Jack & Pearl Resnick Campus 1300 Morris Park Ave. Bronx, NY 10461 (718) 430-2890 Fax: 430-8996 URL: http://www.aecom.yu.edu/aif/
Dear Listeners, I am making ferrite nanoparticles doped with Co and/or Mn in the size range of 20-100nm. I have looked at them with TEM and made up size distributions from the pictures. However, I would now like to check my distribution curves with a particle sizer. Does anyone now if this is possible? I suspect it to be difficult since my particles are magnetic. If it is possible, does anyone know what type of medium I can disperse my particles in? So far I have tried to alter the pH but it doesn't seem to be enough and the particles still agglomerate before the the measurement is over. Thanks .
Yours sincerely Richard
PhD student, Swedish Defense Research Agency Department of Polymer Technology Royal Institute of Technology Teknikringen 56 SE-100 44 Stockholm SWEDEN phone: +46-8-790 76 37
--------------------------------------------------------- Legal Notice: This electronic mail and its attachments are intended solely for the person(s) to whom they are addressed and contain information which is confidential or otherwise protected from disclosure, except for the purpose they are intended to. Dissemination, distribution, or reproduction by anyone other than their intended recipients is prohibited and may be illegal. If you are not an intended recipient, please immediately inform the sender and send him/her back the present e-mail and its attachments and destroy any copies which may be in your possession.
Our webppage on connecting a Coolpix to a microscope will answer a lot of your questions: http://www.mvia.com/Coolpix/clpxadpt.htm
Also, please see my answers below...
Thanks! Jim Haley
****************************** Jim Haley Applications Engineer MVIA, Inc. 125 Sherwood Drive Monaca, PA 15061 voice: (724) 728-7493 fax: (412) 291-1709 e-mail: haley-at-mvia.com webpage: http://www.mvia.com ******************************
Alan Davis wrote: } } The Coolpix 5000 looks interesting. It does much of what I need, } including monitoring the live specimen on a TV, Computer, or video } projector. I like some of the features of the 5700, including better } telephoto range for shore bird studies. I have a couple of questions I } would refer to those with Coolpix experience: } } How well does the 5700 work on a scope?
Very poorly - you'll need to use the digital zoom mode on the 5700. See our FAQ section: http://www.mvia.com/Coolpix/clpxadpt.htm#FAQ
} Does the 5700 identically to the 5000 out of the box, with the Nikon } adaptor kit?
NO! You'll need to use the digital zoom mode on the 5700 as opposed to the optical zoom mode on the 5000, which will result in very poor images.
} Are these cameras significantly better than the 4500?
The 5000 has a higher resolution, but does not have a swivel body as the 4500 model does, which make it a little less ergonomical. However, the 500 will give you better images.
} Are macro kits available to enable focus to within 1/8"? (I intend } to use it as an "aquarium microscope" for example to study the community } at the sediment/water column interface, for which the Videoflex camera } works admirably well.)
There are a couple of lenses available from Nikon for various wide angle and fisheye applications.
} Can one take a shot easily an within ordinary lag values while } monitoring in real time?
Yes, but the live image will pause when you capture your image.
} Nikon has a relay lens (is that a projection lens?) for this } purpose, that will work on Coolpix 5000; does it work ok on the 5700? } } Another question: is the Nikon D-100 worth the extra cost, and does it } incorporate the video monitoring Coolpix features? Is it useable on a } scope, and how? } } Thank you for any comments or suggestions. I apologize for asking so } many questions at once... } } Alan Davis } Marianas High School } Saipan } } I wish to ask whether microscopists feel it } } -- } adavis-at-saipan.com 1-670-322-6580 } Alan E. Davis, PMB 30, Box 10006, Saipan, MP 96950-8906, CNMI } } I have steadily endeavored to keep my mind free, so as to give up any } hypothesis, however much beloved -- and I cannot resist forming one } on every subject -- as soon as facts are shown to be opposed to it. } -- Charles Darwin (1809-1882) } } The right to search for truth implies also a duty; one must not } conceal any part of what one has recognized to be true. } -- Albert Einstein } } As we enjoy great advantages from the inventions of others we should } be glad of an opportunity to serve others by any invention of } ours, and this we should do freely and generously. } -- Benjamin Franklin } }
Here is our answer to the many responses concerning the previous thread on Torr Seal. Torr Seal is an extremely high quality epoxy resin manufactured under a private label agreement as Torr Seal. Since there are non-critical applications which do not require such a high quality epoxy we, like others do carry a product known as a Torr Seal equivalent. We do let all our users know that when they select an 'equivalent they are paying less, but naturally there is a sacrifice in quality.
John Arnott
Disclaimer: Ladd Research sells the products mentioned in this e-mail.
Ladd Research 83 Holly Court Williston, VT 05495
On-line Catalog: http://www.laddresearch.com
tel: 1-802-658-4961(anywhere) or 1-800-451-3406(US) fax: 1-802-660-8859 e-mail: sales-at-laddresearch.com
Dear Listeners, I am making ferrite nanoparticles doped with Co and/or Mn in the size range of 20-100nm. I have looked at them with TEM and made up size distributions from the pictures. However, I would now like to check my distribution curves with a particle sizer. Does anyone now if this is possible? I suspect it to be difficult since my particles are magnetic. If it is possible, does anyone know what type of medium I can disperse my particles in? So far I have tried to alter the pH but it doesn't seem to be enough and the particles still agglomerate before the the measurement is over. Thanks .
Yours sincerely Richard
PhD student, Swedish Defense Research Agency Department of Polymer Technology Royal Institute of Technology Teknikringen 56 SE-100 44 Stockholm SWEDEN phone: +46-8-790 76 37
--------------------------------------------------------- Legal Notice: This electronic mail and its attachments are intended solely for the person(s) to whom they are addressed and contain information which is confidential or otherwise protected from disclosure, except for the purpose they are intended to. Dissemination, distribution, or reproduction by anyone other than their intended recipients is prohibited and may be illegal. If you are not an intended recipient, please immediately inform the sender and send him/her back the present e-mail and its attachments and destroy any copies which may be in your possession.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (Jlindstrom-at-Hotmail.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Tuesday, September 2, 2003 at 17:19:35 ---------------------------------------------------------------------------
Email: Jlindstrom-at-Hotmail.com Name: james lindstrom
I would appreciate off-list feedback from users of JEOL 6700F tools.
I'm looking at a system with turbo pump, large specimen exchange interlock, solid state BSE vs. Robinson BSE, beam stability, accommodation to EDAX Genesis EDS and overall usability and support. Location is Northern CA.
Off-list please. All are confidential. Trying to make a purchase decision between this and comparable Hitachi. Specimens are microchips in cross section, top down, metallurgical coupons of IC manufacturing tools, legacy support for Amray 12mm diameter 3.1mm pin specimen stubs. Also, there is a need for working with rather large specimens. Like blocks of NIST standards (100mm diameter, 50mm high).
Native digital capture resolution is low. Are there other options that work? Is anyone using Soft Imaging ADDA? 4Kx4K pixels minimum is needed.
} From: "Michael Cammer" {cammer-at-aecom.yu.edu} : : } not related to the real safety (mostly keeping paperwork in a good shape : } and accurately fill out chemical waste disposal tags). So, in such : } situation, I think, it's superviser's personal responsibility to provide : } safely environment to the workers and train them accordingly. : : Look, we don't want to have this devolve into a session on what is safe and : what isn't, but this is a very simplistic view. Paperwork may be essential : for real safety of all handlers of material down the chain of disposal. If : a supervisor has a limited budget, he/she is going to scrimp on safety. : : Policies are in place to protect workers who historically have been : mistreated. Yes, blanket policies do not apply in all situations. The : bureaucracy simply needs some flexibility for individually documented : exemptions. : : } P.S. By the way: in my EM work, my glasses, I wear to correct my vision, : } save my eyes in uncounted number of times. I, also, used to remove them : } every time I am using light microscope or binoculars. I don't see big : } problem removing them if necessary. But: yes, it decreases my : } productivity. Ok, let say, I need about 5 sec. to remove them and put : } back. Let say I do it 40 times a day: 5 secx40=200 sec/3.3 min per day. : : But you're wearing gloves and your gloves shouldn't be near your unshielded : eyes each time you have to handle the googles. So you really should take : off your gloves before removing your goggles each time. : Face shields will solve the glove problem. If the curve of plastic face shields interferes with your work a welders helmet with plain glass or you prescription lenses could be used. You would need some clearance behind the microscope but it would speed things up and not run the risk of getting something in your eye when you were removing goggles. With prescription lens in the helmet it should speed up work as you don't have to remove your glasses. It also solves the problem of scratched face shields.
Here is an example http://cgi.ebay.com/ws/eBayISAPI.dll?ViewItem&item=2551210520&category=633 The part below the lens should be cut away as you don't need protection from the burning rays of the welders arc and the dark lens replaced with a clear one. Then just the push of your thumb will stand the helmet straight up out of the way of a microscope.
A welders helmet should be sturdy enough to satisfy any safety officer and quicker to tip up than removing goggles. Some surgery on the hood would make it lighter and reduce the clearance need behind the microscope with out compromising eye safety.
There are also some goggles that have a flip up lens available at welders supply houses as well. I don't know if they would clear a microscope. Here is an example http://cgi.ebay.com/ebaymotors/ws/eBayISAPI.dll?ViewItem&category=35000&item=2429373537 There are other designs that might be more suitable with the lens swinging 180 degrees. A call to your local Air Gas or other welding supply dealer will let you know what is available.
Gordon Couger gcouger-at-couger.com
I collect links on information related to light microscopes. http://www.couger.com/microscope/links/gclinks.html Please forward any links or information you think might be useful to others.
Dear Colleagues, We have following trouble with Zephyr ZEM 1000CW watercooling unit: When our CM12 is switched on from STAND BY, overload protector switches off the pump motor of ZEM 1000CW from time to time. If the reset button of overload protector is pressed down, the motor starts and all is OK. But after several days or weeks this occurs again. Please, can you give us some hints or suggestions. Many thanks in advance. Oldrich Benada
+-----------------------------------+ Oldrich Benada Acad. Sci. CR Institute of Microbiology Laboratory of electron microscopy Videnska 1083 CZ - 142 20 Prague 4 - Krc Czech Republic +------------------------------------+ Phone: +420-241062399 Fax: +420-241062347 WEB: http://www.biomed.cas.cz/mbu/lem113/lem.htm
Hello, I am searching for a manual for Reichert FC4 cryoultramicrotome. Is there anybody who could help me with that? Best regards,
Dr Dobroslawa Budka Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa fax: +27-21-8433543 phone: +27-21-8431161 mobile phone: 0722656091
We are looking for volunteers to perform beta testing of some code to do HREM image processing and Direct Methods. The code is a gnu-like package (i.e. free) which should compile on any unix or Mac computer fairly simply using a conventional "./configure ; make ; make install" strategy. If you know what I am talking about and are interested, please email me (not the whole list). Thanks.
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm2-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
I just about wrote "Urgent Assistance Needed" as the subject line, but then realized that would probably not make it past several dozen spam filters. I'm in my usual position (stuck between a rock and a deadline). I'm looking for a copy (original reprint or the journal issue) of:
MANTON I. & VON STOSCH H.A. 1966. Observations on the fine structure of the male gamete of the marine centric diatom Lithodesmium undulatum. Journal of the Royal Microscopical Society 85: 119-134.
I need to scan several figures from the paper, and our interlibrary loans can only get a xerox copy. Can anybody help? I'd gladly pay courier charges (both ways if you need the journal/paper back). Alternatively, if someone is willing, they could scan the figures from their end, but it would most likely be all of the plates in the paper, as I'm not sure right now which ones I need.
If you help, I'll make several offerings on your behalf to the gods of microscopy, requesting long filament life, long strings of buttery smooth silver-gray thin sections, and awesome powers against the Dark Forces of Bureaucracy. I'll also put you on the list to receive the coveted large format scan of the Philips 200 column cross-section poster (discussed earlier in this forum) which I should have constructed and cleaned up Real Soon Now.
Please contact me off-list. Thanks,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
I'm trying to stain liver sections embedded in paraffin and fixed by metacarn fixative. My first antibody is monoclonal. The problem is that I only have a positive signal at the periphery of the section and nothing in the middle. Does someone have any clue why this could happen? Thanks in advance.
Prof. Dr. Francisco Javier Hernandez Blazquez Universidade de São Paulo Fac. de Medicina Veterinária e Zootecnia Departamento de Cirurgia - Setor de Anatomia Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
Thanks for the quick reply, Chris (and others who may be in the pipe),
Even my good luck seems to have a bad side! Not 5 minutes after posting this request, I found a copy of this paper, literally under my nose. Embarassing to say the least! I'll be quiet for at least a week....
Cheers,
Jim
Christopher F. Blanford wrote:
} James - } } Unfortunately we can't take any copies of the journal out of the } library. But as the RMS is just up the street from me, do you think } you could just give them a call? Either see if they have a copy they } could send you, or if you're still in a bind, see if they'd loan me a } copy for a day and I could scan in the pages you want. The number: +44 } 1865 248 768. } } Good luck. } } Chris } } On Wednesday, September 3, 2003, at 01:40 pm, James M. Ehrman wrote: } } } Hi all, } } } } I just about wrote "Urgent Assistance Needed" as the subject line, } } but then realized that would probably not make it past several dozen } } spam } } filters. I'm in my usual position (stuck between a rock and a } } deadline). I'm looking } } for a copy (original reprint or the journal issue) of: } } } } MANTON I. & VON STOSCH H.A. 1966. Observations on the fine structure } } of the male gamete of the marine centric diatom Lithodesmium } } undulatum. Journal of } } the Royal Microscopical Society 85: 119-134. } } } } I need to scan several figures from the paper, and our interlibrary } } loans can only } } get a xerox copy. Can anybody help? I'd gladly pay courier charges } } (both ways } } if you need the journal/paper back). Alternatively, if someone is } } willing, they could } } scan the figures from their end, but it would most likely be all of } } the plates in the paper, } } as I'm not sure right now which ones I need. } } } } If you help, I'll make several offerings on your behalf to the gods } } of microscopy, requesting } } long filament life, long strings of buttery smooth silver-gray thin } } sections, and awesome } } powers against the Dark Forces of Bureaucracy. I'll also put you on } } the list to receive } } the coveted large format scan of the Philips 200 column cross-section } } poster (discussed } } earlier in this forum) which I should have constructed and cleaned up } } Real Soon Now. } } } } Please contact me off-list. Thanks, } } } } Jim } } } } -- } } } } James M. Ehrman } } Digital Microscopy Facility } } Mount Allison University } } Sackville, NB E4L 1G7 } } CANADA } } } } phone: 506-364-2519 } } fax: 506-364-2505 } } email: jehrman-at-mta.ca } } www: http://www.mta.ca/~jehrman } } } } } } } } } }
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (taylor-at-bio.fsu.edu) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 3, 2003 at 11:21:33 ---------------------------------------------------------------------------
Email: taylor-at-bio.fsu.edu Name: Kenneth Taylor
Organization: Florida State University
Title-Subject: Fixation and Embedding of bacteria
Question: Hi,
I am looking for a protocol for fixing, embedding and sectioning bacteria, in particular E. coli. Good membrane preservation would be important for this study.
Does anyone have a protocol that they would be willing to share?
OK...thanks to all of the respondents about FESEMs. I did receive some good feedback.
I am trying to make a budget wedge for FY04 and need to know a basic idea of benefits of what I have selected as candidate systems. That said, I also need to put in funding wedges to cover the venue of appropriate SEMs.
JEOL 6700F Hitachi S-4800 Hitachi S-5200
Other options are welcome too.
I would appreciate knowing personal experiences based on differing sizes of specimens and different types of analysis. Budget time comes along one time, and is perversely ignorant of systems. Caveat emptor.
It may be time to upgrade. But to what? Cold FE with accommodation of my EDAX Cryospec is necessary. Robinson BSE is also a big plus.
Hello, could you please let me know who the suppliers of HISTORESIN are. It is a glycol methacrylate usful for light microscopy. It used to be marketed originally as LKB HISTORESIN. Thank you, Dobrusia
Dr Dobroslawa Budka Materials Research Group iThemba LABS PO Box 722 Somerset West 7129 South Africa fax: +27-21-8433543 phone: +27-21-8431161 mobile phone: 0722656091
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (reckert-at-nwlabs1896.com) from http://www.msa.microscopy.com/MicroscopyListserver/MLFormMail.html on Wednesday, September 3, 2003 at 21:24:36 ---------------------------------------------------------------------------
Question: We are interested in purchasing a CamScan CS44 SEM (appr. 10 years old) and I was wondering if anybody could share their experience (weak and strong points) about the scope. Thank you.
Well, BILLPC has stopped sending me copies of a virus, but I still get copies from PRIVAT (pD9E76707.dip.t-dialin.net [217.231.103.7], and MINIHOTEL (modemcable027.214-131-66.nowhere.mc.videotron.ca [66.131.214.27]
Some of the spoofed addresses/domains on the messages are ones I recognize from this list. So if any of you recognize these folks or their domains, would you please tell them to check out their machines. Thanks.
Hello Historesin is now produced and sold by Leica Microsystems Nussloch GmbH. Their address is Heidelberg Strabe 17-19 D-69226 Nussloch/Heidelberg Tel. 0 62 24/143-0 You may look at their homepage at http://www.histo-solutions.com/website/sc_hbu.nsf
Good luck
Prof. Dr. Francisco Javier Hernandez Blazquez University of São Paulo Fac. Veterinary Medicine and Animal Sciences Departament of Surgery - Sector of Anatomy Av. Prof. Dr. Orlando Marques de Paiva, 87 05508-000 - São Paulo (SP) - Brasil Tel..55 (11) 3091 1374 Fax 55 (11) 3091 7805 email: fjhblazq-at-usp.br
----- Original Message ----- } From: "dobrusia budka" {dobrusiabudka-at-tlabs.ac.za} To: "Microscopy Listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Thursday, September 04, 2003 8:26 AM
Hi:
Help!!! I'm presently having problems with photography of nerve tissue. My problem is that the tissue appears shredded, and it has multiple holes and splits along the tissue but not the plastic. Making the tissue impossible to photo. This problem can only be seen when we are to photograph in the microscope. 1 micron light slides look fine. The only thing we have change is the type of fixative. We went from glutaraldehyde to para formaldehyde, but nothing have change from my protocol. We think is an infiltration problem, because the tissue is big to start and humidity have been a problem in the past in other types of tissue, and because the holes and splits are only in the tissue. We are looking for processing protocols for Sural Nerve tissue in order to compare with the one we have. If anyone can help it will be greatly appreciated.
Thanks
Omayra Velez MS Electron Microscopy Specialist Pathology Department Weill Cornell Medical College 1300 York Ave NY,NY, 10021 212-746-6437 omv2001-at-med.cornell.edu
__________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com
Does anyone know anything about the ISI WB-6 that is currently being offered on eBay? A friend of mine was wondering about the specifics concerning the condition and usability of the instrument.
Omayra It looks like fixation/impregnation problem. Paraformaldehyde itself is not enough for EM. I know nothing about your particular sample, in general, we are using 4% paraformaldehyde+ 1% GA overnight (+4oC). Nervous tissue also contains a lot of lipids, so dehydratation should be slow: actually, all steps should be longer than usual. I do find that Spurr resin is good for brain tissue: it penetrates better than Epon or Araldite. Good luck, Sergey.
At 02:04 PM 9/4/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry 10833 Le Conte Ave, Room 33-089 Los Angeles, CA 90095
Historesin as such is now discontinued but lives on as a series of resins produced by Heraeus Kulzer in Germany - Technovit 7100 (Historesin), Technovit 8100 (Historesin Plus) and Technovit 9100 NEW (Histodur).
These resins are all identical to the Historesin products. TAAB Laboratories Equipment Ltd is the Distributor for these products in the UK, but if anyone has a problem finding the distributor in their country, I may be able to help,
Best regards
Terry Cooper TAAB Laboratories Equipment Ltd 3 Minerva House, Calleva Park Aldermaston, Berks, RG7 8NA, England Tel ++44 (0)118 981 7775 Fax ++44 (0)118 981 7881 e-mail sales-at-taab.co.uk www.taab.co.uk
I have problems with my EBSD measurements. The image of the phosphorscreen is distorted at the bottom of the screen while substracting the backgroung. instead of pure cirlces at the bottom you see more or less straight lines cutting the circles or the circles themselfs are distorted. When substracting at very low magnifiocations this problem doesnŽt occure. When you look at the RAW picture the area at the bottom shows a dark line. Please I need help. It is urgent
Regards Dirk Kirch -- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
I have problems with my EBSD measurements. The image of the phosphorscreen is distorted at the bottom of the screen while substracting the backgroung. instead of pure cirlces at the bottom you see more or less straight lines cutting the circles or the circles themselfs are distorted. When substracting at very low magnifiocations this problem doesnŽt occure. When you look at the RAW picture the area at the bottom shows a dark line. Please I need help. It is urgent
Regards Dirk Kirch
-- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
-- begin:vcard n:Kirch;Dirk tel;fax:+49(0)241-8022301 tel;work:+49(0)241-8026861 x-mozilla-html:FALSE url:www.imm.rwth-aachen.de org:Institute of Physical Metallurgy and Metal Physics;University of Aachen version:2.1 email;internet:kirch-at-imm.rwth-aachen.de title:Dipl.- Phys. Dirk Kirch adr;quoted-printable:;;Kopernikusstra=DFe 14=0D=0A;52056 Aachen;Germany;; end:vcard
We are thinking to buy a liquid helium cooled double tilt TEM holder. Has anyone used this type of holder? Can you please tell me your experience with it? I would appreciate very much if you could tell me how well it works, and what problems it might have.
I would like to ask more specifically about the liquid helium cooling TEM holder made by USA side of the Gatan company. The UK made holder will be too expensive for us, so anyone has experience with the US made holder?
Hi to all: Thanks for the responce on the nerve problem. Some of you have asked how big the tissue samples are and the type of resin I'm using. We are using, EMBED 812 with DMP-30 as catalyst. The tissue size is about 5mm x 2mm x 3mm. Big for the most part. I already tryed to make the blocks smaller, but the nero-pathologist wanted the pieces this big. We have a 20 hour protocol from buffer to 100% epon. I'll try some of the sudjestions, lenthtening the protocol. Although we think we are also having a humidity issue as well, due to construction and AC problems.
Thanks again
Omayra Velez Electron Microscopy Specialist Pathology Department Weill Cornell Medical College 1300 York Ave NY,NY. 10021 212-746-6437 omv2001-at-med.cornell.edu
__________________________________ Do you Yahoo!? Yahoo! SiteBuilder - Free, easy-to-use web site design software http://sitebuilder.yahoo.com
Here is the table of contents for the September/October 2003 issue of Microscopy Today.
New Subscriptions via http://www.microscopy-today.com only, please New subscriptions will close on Tuesday 9 September for this issue.
There has been a major change in subscription policies coming out of the MSA Winter Council meeting. Briefly: Canadians and Mexicans are now offered free subscriptions along with microscopists in the USA. MSA members anywhere have free subscriptions. Non-MSA, non-North American subscriptions have been reduced from $80 or $110US to $35US. Additional details in the magazine or on our web site.
Table of Contents:
Carmichael Print Your Own Organs! Kelly and Gribb Atom Probes LEAP Ahead Breger Microscopy At The Ends Of The Earth Barkan, et al. A Si Multi-Cathode Detector For Microanalysis Applications Sehgal, Karim, Stafford, & Fasolka Techniques for Combinatorial and High- Throughput Microscopy Part 1: Gradient Spec. Fabrication for Polymer Thin Film Research Millette, Boltin, Few, & Turner Microscopical Studies of World Trade Center Disaster Dust Particles Vrdoljak Prep. Of Soil Samples For Light And TEM Radzikowska A New Look At Cast Iron Microstructure Lifshin & Gauvin Precision and Detection Limits for EDS Analysis in the SEM Sedgewick Adding Color To Grayscale Images Downing Support Films with Uniform Hole Size Mills Cleaning a Cold Cathode Gauge Tube Yang and Kalab Zigzag Edges in SEM Micrographs Schooley Microscopy CD-ROMs For Children: A Bibliography
Does anyone have a package or know of a package that can calculate changes in sample thickness from thickness fringes observed in conventional TEM? I am imageing defect free single crystals on the order of 100nm. I need to know how the crystals thickness profile changes.
I have a student who wants to image magnetic particles with SEM. The particles are in the 400nm to 1µm size range. I had planned to stick them to double stick copper tape prior to imaging. However, I am concerned about their adversely affecting the microscope. Are there any suggestions as to mounting the particles, whether short working distance may magnify potential problems if some particles break loose and get into the lens. Etc?
If there is a concern then I guess it would be possible to mount on sticky carbon tape, overlay the sample with a formvar film, and then use backscattering to image the particles. However, is this necessary?
Debby
Debby Sherman, Manager Phone: 765-494-6666 Life Science Microscopy Facility FAX: 765-494-5896 Purdue University E-mail: dsherman-at-purdue.edu S-052 Whistler Building 170 S. University Street West Lafayette, IN 47907
If one wants to do pure standards-based quantitative EDS analysis, protocol says that conditions for the standards must be the same as for the specimen. Well, if one is using a thermal FE SEM, then the beam current is very likely to be the same at any time. How about for a cold FE gun?
From what I can surmise, the cold guns vary much more than thermal FE guns, to the point that intervention is necessary. Does this mean that every quant EDS measurement has to have a precursor Faraday cup reading of beam current? Conversely, does this mean that this is not required when using a thermal FE gun?
The EDAX system supports a Beam Current Factor so that calculations are made based on as-read current when making the quant analysis. This seems like a lot of hassle. Is the difference between standardless and standards-based EDS so huge that the hassle is warranted? Or, is this nit picking?
Is it possible to simply degauss the samples before putting them in the SEM?
There is a border, I believe, between too much degaussing and too little. Looking at hard drive tracks is a case in point. Depending on your ultimate resolution, the working distance may facilitate your specimens by keeping them away from the final lens. 1um specimens should not require short WD.
Your BSE ought to be a separate issue depending on what type it is and what KV it works at.
gary g.
At 03:14 PM 9/5/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
We have routinely examined (gamma-)ferric oxide particles in the SEM by dusting the particles onto double-sided sticky carbon tape. Or you can paint a small dab of carbon paint onto your stub and dust the particles onto the paint while it is still wet. Then tap the side of the stub firmly on a hard surface to dislodge any loose particles. If you are really paranoid you can use a low-pressure gas stream (like a "Micro Duster") to make sure there are no loose particles present. However, the more aggressively you remove looser particles, the less you are likely to see larger particles that may exist in the specimen. This may be important if you are trying to do particle size distribution measurements.
Particles like Fe2O3 are weakly magnetic, so there is not much danger of them being sucked up into your lenses by the magnetic field.
You will probably want to keep the working distance fairly short ( {10mm) to achieve acceptable image quality.
Hope this helps, Ken Gaugler
Debby Sherman wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi all, } } I have a student who wants to image magnetic particles with SEM. The } particles are in the 400nm to 1µm size range. I had planned to stick them } to double stick copper tape prior to imaging. However, I am concerned about } their adversely affecting the microscope. Are there any suggestions as to } mounting the particles, whether short working distance may magnify potential } problems if some particles break loose and get into the lens. Etc? } } If there is a concern then I guess it would be possible to mount on } sticky carbon tape, overlay the sample with a formvar film, and then use } backscattering to image the particles. However, is this necessary? } } Debby } } Debby Sherman, Manager Phone: 765-494-6666 } Life Science Microscopy Facility FAX: 765-494-5896 } Purdue University E-mail: dsherman-at-purdue.edu } S-052 Whistler Building } 170 S. University Street } West Lafayette, IN 47907 } } }
Wouldn't it also be dependent on the type of SEM used (i.e. HRSEM)?
If the final lens (pole piece) is not energized for through-the-lens detection, the sticky Cu tape should hold them down. Disperse the particles then blow off the weakly held particles. Now lower a ferro magnetic material over the samples and see if any pop off. If particles do "pop" off then use formvar and a high kV. The magnetism of the particles may play a role in their physical construct so degauss with caution. I believe imaging aberrations will most likely be the issue over that of tool contamination. I have successfully imaged toner particles in a relatively low resolution SEM. It should be noted that this tool has been classified as a "dirty" tool. The toner particle were even dispersed on the "new" C sticky tabs. The ones that are rock hard with barely any adhesive.
Kristopher
-----Original Message----- } From: Gary Gaugler [mailto:gary-at-gaugler.com] Sent: Friday, September 05, 2003 7:28 PM To: Debby Sherman Cc: MSA listserver
Is it possible to simply degauss the samples before putting them in the SEM?
There is a border, I believe, between too much degaussing and too little. Looking at hard drive tracks is a case in point. Depending on your ultimate resolution, the working distance may facilitate your specimens by keeping them away from the final lens. 1um specimens should not require short WD.
Your BSE ought to be a separate issue depending on what type it is and what KV it works at.
gary g.
At 03:14 PM 9/5/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
You really don't need a package. You can do this by hand using basic TEM techniques.
I think that your best way to do this would be to set up a good 2-beam condition and use convergent beam electron diffraction. You use the distances between the symmetrical fringes (Kossel-Mollenstedt fringes) in the known reflection, calculate the deviation from exact Bragg condition for each fringe, and plot the results assuming that the first fringe that you see is the 1st, 2nd, or 3rd. You plot s(i)^2/n(i)^2 vs 1/n(i)^2, where n(i) is the index of the particular fringe. You try different starting values for n(1) starting from 1, then 2, then 3. You usually don't have to go higher than 3. When you get a straight line, the slope of the straight line is -1/(extdist)^2 and the intercept of the line is 1/t^2. This comes from the following relationship,
s(i)^2/n(i)^2 + 1/(extdist)^2/n(i)^2 = 1/t^2.
This technique is demonstrated in a number of TEM texts. The procedure is written in the Williams and Carter book.
You could apply this at two points on the sample along the wedge and get the angle.
You could also do this from the results of the two-beam approximation model. If you are at 2-beam Bright Field condition and you do not have bend contours present, then the first dark fringe that occurs in the wedge will occur at a thickness of 0.5 * (extdist(g)) and the second at 1.5 * (extdist(g)), (and so forth.) From the lateral separation distance between the fringes and the value of the extinction distance for that reflection, you can calculate the wedge angle. This is also written up in the Williams and Carter book and others as well.
-Scott
Scott D. Walck, Ph.D. PPG Industries, Inc. Glass Technology Center P. O. Box 11472 (letters) Guys Run Rd. (packages) Pittsburgh, PA 15238-0472
Walck-at-PPG.com
(412) 820-8651 (office) (412) 820-8515 (fax)
-----Original Message----- } From: thurston e herricks [mailto:thurst0n-at-u.washington.edu] Sent: Friday, September 05, 2003 3:37 PM To: Microscopy-at-sparc5.microscopy.com
Hello all,
Does anyone have a package or know of a package that can calculate changes in sample thickness from thickness fringes observed in conventional TEM? I am imageing defect free single crystals on the order of 100nm. I need to know how the crystals thickness profile changes.
Take care
Thurston Herricks
MicroscopyListserver Archive Email Extraction Software Version NJZ07060908