A student would like to bring back phage and bacterial samples from Mexico to the UK.
With increased security at airports, he was considering bringing the pellets back in a sucrose/water solution rather than sucrose/cacodylate. The samples would be in this medium for approx. 15hours, and would be fixed in 2.5% Gluteraldehyde in 0.1M Sodium Cacodylate buffer. I am a bit concerned about leaving the samples in the sucrose/water medium, but what is the alternative?
Where he is going has no EM facilities, although they can obtain reagents.
Any advice would be greatly appreciated.
Linton Brown Institute of Aquaculture University of Stirling Stirling FK9 4LA Scotland UK -- The University of Stirling is a university established in Scotland by charter at Stirling, FK9 4LA. Privileged/Confidential Information may be contained in this message. If you are not the addressee indicated in this message (or responsible for delivery of the message to such person), you may not disclose, copy or deliver this message to anyone and any action taken or omitted to be taken in reliance on it, is prohibited and may be unlawful. In such case, you should destroy this message and kindly notify the sender by reply email. Please advise immediately if you or your employer do not consent to Internet email for messages of this kind. Opinions, conclusions and other information in this message that do not relate to the official business of the University of Stirling shall be understood as neither given nor endorsed by it.
Dear all, Thank you to all who replied and made suggestions about how we could fix things with the misaligned JEOL3010. The suggestion to use the "N" button and recover the engineers settings was a good one. I still didn't have much beam afterwards, but it was then close enough to adjust normally. The worst adjusted bits were the gun deflectors and condensor deflectors. The adjustment was made harder since the LAB6 crystal is a bit tilted to one side, but I have a good workable alignment now (even if at some point we have to look at the filament itself if it continues to tilt off).
So, thanks once again to all.
Best wishes
-- Ian MacLaren Technische Universität Darmstadt Material-und Geowissenschaften Petersenstr. 23 64287 Darmstadt Germany
This is an issue with many answers, depending on the type of work preformed.
Are you a quality control lab that is spot checking production type samples? If so, than a known defective product is given to the employee and asked to find any problems.
If you are a diagnostic TEM lab, than, are they responsible for actually making the diagnosis? Probably not, thus checking the quality of thicks and thins is the only objective way to due quality control. Look for useable thicks with minimal cutting and staining artifacts. When checking the thins, look for section quality, staining artifact and final micrograph quality. One can objectively review these items.
On the SEM, are you doing ED or WD x-ray analysis? Here a know sample can be used as a test material to determine if the proper x-ray spectrum is being determined. If bio. samples, look for CPD artifacts, such as ruptured membranes.
One way to help insure quality (if a bio. lab) is too have the techs become certified by MSA, thru their certification program.
This is not an easy answer, but if you can objectively critique the work on a daily basis, quality control should not be an issue.
Best of Luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} 03/31/03 07:42AM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Good morning all in the listserver, I had a question - how does on go about doing quality control on EM - both TEM and SEM. In other words, other than giving techs unknown samples and checking the end results - or just looking over their shoulders checking thicks and thin quality - is there another way? Thanks in advance, Connie Cummings
Why look for another service company, Philips is willing to work on a pay for service rate. Check with them, that way you still can maintain the current service person.
Best of Luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } Rick Harris {raharris-at-ucdavis.edu} 03/31/03 05:35PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As part of our restructuring, we will not renew the service contract on our trusty old Philips410LS. Can anyone recommend third party service in Northern California? I understand Ron Veil is no longer in business.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
} -------- } From: Linton Brown } Sent: 01 April 2003 12:09 } To: 'Microscopy-at-MSA.Microscopy.Com' } Subject: Transporting samples } } A student would like to bring back phage and bacterial samples from Mexico } to the UK. } } With increased security at airports, he was considering bringing the } pellets back in a sucrose/water solution rather than sucrose/cacodylate. } The samples would be in this medium for approx. 15hours, and would be } fixed in 2.5% Gluteraldehyde in 0.1M Sodium Cacodylate buffer. I am a bit } concerned about leaving the samples in the sucrose/water medium, but what } is the alternative? } } Where he is going has no EM facilities, although they can obtain reagents. } } Any advice would be greatly appreciated. } } Linton Brown } Institute of Aquaculture } University of Stirling } Stirling } FK9 4LA } Scotland } UK } } -- The University of Stirling is a university established in Scotland by charter at Stirling, FK9 4LA. Privileged/Confidential Information may be contained in this message. If you are not the addressee indicated in this message (or responsible for delivery of the message to such person), you may not disclose, copy or deliver this message to anyone and any action taken or omitted to be taken in reliance on it, is prohibited and may be unlawful. In such case, you should destroy this message and kindly notify the sender by reply email. Please advise immediately if you or your employer do not consent to Internet email for messages of this kind. Opinions, conclusions and other information in this message that do not relate to the official business of the University of Stirling shall be understood as neither given nor endorsed by it.
Hi Does anyone know when the word "EDAX" was registered as a trademark? The reason I ask is that I have come across procedural documentation that calls for "...analysis by EDAX (Energy Dispersive Analysis of X-Rays)." One lab is now telling us that they cannot comply as the don't have EDAX - theirs is a PGT system !
Ady
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EDAX is the name of a company which provides EDXA (or EDS or EDX or...) equipment. They were founded in 1962 as Nuclear Diodes Inc. and have operated under the name EDAX since 1972. (see www.EDAX.com) EDAX is a tradename just as "Kleenex" is for facial tissue or "Xerox" is for photocopies.
A number of companies manufacture EDX equipment including: EDAX, Gresham, IXRF, Noran, Oxford, PGT, Rontec, ... (I may have missed some.)
There have been a number of discussions about the "proper" terminology for the Energy Dispersive method of X-Ray Analysis. There are even quibbles about the term "energy-dispersive".
The writer of your procedures was not careful enough about his terminology.
Cheers, Henk
At 06:37 AM 4/1/2003 -0800, ady jenkinson wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Hendrik O. Colijn colijn.1-at-osu.edu Campus Electron Optics Facility Ohio State University (614) 292-0674 http://www.ceof.ohio-state.edu Time is that quality of nature which keeps events from happening all at once. Lately it doesn't seem to be working.
I have co-written a paper on "Quality in Electron Microscopy" which I would be pleased to send through to you if you are interested?
It deals with test procedures, instrument and staff assessment, quality control in an EM unit etc etc.
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: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, March 31, 2003 1:42 PM
Announcing an Additional Sunday Short Course Scheduled for Microscopy and Microanalysis 2003 San Antonio TX August 3, 2003
Application Pathways Solutions: From High Pressure Freezing to the Electron Microscope
Instructors: Kent McDonald Jan Slot
Location: San Antonio Convention Center, Room 008B Time: 9:00am to 5:00pm Attendance: Maximum 20 people Attendance Fee: same as other Sunday short courses. Payable with meeting registration or by contacting Meeting Management at:
Workshop outline (Combination of didactic and practical sessions): - Introduction of High Pressure Freezing, Why HPF? - Advantages over conventional chemical and or microwave fixation techniques. - Techniques and methods for Sample preparation after HPF including; - Freeze fracture - Frozen hydrated cryosectioning - Cryosectioning and immunolabeling - Cryoplaning - Freeze substitution. - Additional presentations
For registration information see the meeting site at: http://www.msa.microscopy.com/MSAMeetings/MM03/MMHomePage.html
Best- Jay ________________________________ AKA. W. Gray Jerome, Ph.D., FAHA Pathology Vanderbilt University Medical Center B-2101 MCN 1161 21st Ave, South Nashville, TN 37232-2561 615-322-5530 jay.jerome-at-vanderbilt.edu
Hello Ian, On the keyboard draw of the JEOL 3010 is a neutralize button (marked "N" under a clear plastic cover near the inset "reset" button). When this button is pressed the computer resets the presently selected alignment parameter to the last value set by the JEOL service technician when they last aligned your TEM in service mode.
If a user has put your alignment is some bizarre "wopper-jawed" state, pressing the neutralize button will return the setting to the last JEOL alignment. To accomplish this, press an alignment select button in the keyboard drawer then hit the neutralize button. Do this for all the alignment select buttons, e.g. Deflectors (Gun, Spot, Cond, Image, Proj), the Stimators, and Con Def Adj.
This should put your TEM in a useable state if gross misalignment is your only problem. You will need to fine tune the alignment after you neutralize the settings since the operational state of the instrument may have drifted since the last technician stored settings.
I will email you a PDF file of our detailed alignment procedures, which you may find of value.
Good luck and I hope this information helps.
Sincerely, Tyrone Daulton
Ian MacLaren wrote:
} } Dear all, } Our JEOL3010 alignment has got into a bad state, and I can't seem to get it } back with the normal alignments described in the handbook. Probably a user } has accidently altered a gun or condensor alignment. Does anybody have a } set of instructions for a complete gun and column alignment, and not just } the everyday procedures in the standard blue ring-binder? } } } -- } Ian MacLaren } Technische Universität Darmstadt } Material-und Geowissenschaften } Petersenstr. 23 } 64287 Darmstadt } Germany
-- _____________________________________________________ Tyrone L. Daulton Director: Marine Geosciences - Electron Microscopy Facility Physicist and Senior Electron Microscopist
Naval Research Laboratory Marine Geosciences Division (Code 7400) Stennis Space Center, MS 39529-5004
I have researcher gearing up to do a phytolith database (modern and ancient). I have never isolated these myself from fresh tissue and wondered if anyone had any pointers and good references I could look up. Thanks
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
EDAX is a tradename. It is not a trademark. It may have been at one time but not now. You can check for trademarks at
http://www.uspto.gov and do a search of the trademarks database. EDAX returns zero.
Have the procedure revised to kick the A out. EDX. That might work. Or, perhaps EDS?
gary g.
At 06:37 AM 4/1/2003, you wrote:
} Hi } Does anyone know when the word "EDAX" was registered } as a trademark? } The reason I ask is that I have come across procedural } documentation that calls for "...analysis by EDAX } (Energy Dispersive Analysis of X-Rays)." } One lab is now telling us that they cannot comply as } the don't have EDAX - theirs is a PGT system ! } } Ady
Below you will find the information on the EDAX trademark. I think it will answer your questions.
Best regards-
David
Word Mark EDAX Goods and Services IC 009. US 021 026 038. G & S: SCIENTIFIC MEASURING INSTRUMENTS-NAMELY, AN ENERGY DISPERSIVE X-RAY ANALYZER; ANALYTICAL X-RAY APPARATUS, NAMELY X-RAY DETECTORS, X-RAY SPECTROMETERS AND PARTS THEREFOR, ELEMENTAL ANALYZERS, X-RAY GENERATORS, VIDEO DISPLAY CONSOLES FOR X-RAY ANALYSIS, SAMPLE CHAMBERS, REMOTE CONTROLS AND COMPUTER PROGRAMS FOR X-RAY ANALYSIS; COMPUTER-BASED AIDED ANALYTICAL UNITS USING ENERGY DISPERSIVE ANALYSIS. FIRST USE: 19781229. FIRST USE IN COMMERCE: 19781229 Mark Drawing Code (3) DESIGN PLUS WORDS, LETTERS, AND/OR NUMBERS Design Search Code 170725 200308 261701 261704 Serial Number 73453331 Filing Date November 17, 1983 Published for Opposition March 24, 1987 Registration Number 1460660 Registration Date October 13, 1987 Owner (REGISTRANT) EDAX INTERNATIONAL, INC. CORPORATION DELAWARE 103 SHELTER ROAD P.O. BOX 135 PRAIRIE VIEW ILLINOIS 60069 (LAST LISTED OWNER) PHILIPS ELECTRONICS NORTH AMERICA CORPORATION CORPORATION BY CHANGE OF NAME FROM DELAWARE 100 EAST 42ND STREET NEW YORK NEW YORK 10017
Assignment Recorded ASSIGNMENT RECORDED Attorney of Record PAUL R. MILLER Prior Registrations 0925096 Disclaimer NO CLAIM IS MADE TO THE EXCLUSIVE RIGHT TO USE A DESIGN REPRESENTATION OF A GRAPH APART FROM THE MARK AS SHOWN Type of Mark TRADEMARK Register PRINCIPAL Affidavit Text SECT 15. SECT 8 (6-YR). Live/Dead Indicator LIVE
Word Mark EDAX Goods and Services IC 009. US 026. G & S: SCIENTIFIC MEASURING INSTRUMENTS-NAMELY, AN ENERGY DISPERSIVE X-RAY ANALYZER. FIRST USE: 19701000. FIRST USE IN COMMERCE: 19701000 Mark Drawing Code (1) TYPED DRAWING Serial Number 72391254 Filing Date May 6, 1971 Registration Number 0925096 Registration Date December 7, 1971 Owner (REGISTRANT) NUCLEAR DIODES, INC. CORPORATION ILLINOIS 103 SHELTER ROAD PRAIRIE VIEW ILLINOIS 60069 (LAST LISTED OWNER) EDAX, INC. CORPORATION BY CHANGE OF NAME, BY ASSIGNMENT, BY MERGER, BY CHANGE OF NAME, BY MERGER, BY CHANGE OF NAME, BY MERGER, BY ASSIGNMENT DELAWARE 91 MCKEE DRIVE MAHWAH NEW JERSEY 07430
Assignment Recorded ASSIGNMENT RECORDED Attorney of Record CHRISTOPHER R. LEWIS Type of Mark TRADEMARK Register PRINCIPAL Affidavit Text SECT 15. SECT 8 (6-YR). SECTION 8(10-YR) 20020516. Renewal 2ND RENEWAL 20020516 Live/Dead Indicator LIVE
ady jenkinson wrote:
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The problem for EDAX, the company, is that they have not defended their alleged trademark. None of their web pages show that EDAX is a registered trademark. Thus, they are in a minority position. Seemingly, a defensive position. But they may not even know this or be aware of this.
If one has a trademark, one must defend it....or risk losing it. The ® makes a big difference.
gary g.
At 06:39 PM 4/1/2003, you wrote: } -- [ From: Garber, Charles A. * EMC.Ver #3.1a ] -- } Attachment: earl.doc Code: 0KTIJ8M \ Created: 04-01-03, 09:36 PM } [124 Kb] } } I think you are incorrect, the term EDAX is a registered trade name with the } US Office of Patents and Trademarks. } Chuck } } } EDAX is a tradename. It is not a trademark. } } It may have been at one time but not now. } } You can check for trademarks at } } } } http://www.uspto.gov and do a search of } } the trademarks database. EDAX returns zero. } } } } Have the procedure revised to kick the A } } out. EDX. That might work. Or, perhaps EDS? } } } } gary g. } } } } } } } } At 06:37 AM 4/1/2003, you wrote: } } } } } Hi } } } Does anyone know when the word "EDAX" was registered } } } as a trademark? } } } The reason I ask is that I have come across procedural } } } documentation that calls for "...analysis by EDAX } } } (Energy Dispersive Analysis of X-Rays)." } } } One lab is now telling us that they cannot comply as } } } the don't have EDAX - theirs is a PGT system ! } } } } } } Ady } } } } } } -------- REPLY, End of original message -------- }
Reading the postings re JEOL 3010 alignment prompts me to ask whether any kind soul has a procedure for 840 alignment which is easier to follow than the method in the JEOL manual.
I will be grateful for a copy of same.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
John Russ helped start the confusion in terms. He popularized the "EDAX" term with the book "Energy Dispersive Analysis of X-rays" edited by John Russ and published by ASTM in 1971. John then worked for EDAX Inc. in the 1970s before going to North Caroline State.
Do you have any more comments on this history, John?
Ron Vane XEI Scientific
----- Original Message ----- } From: "Hendrik O. Colijn" {colijn.1-at-osu.edu} To: "ady jenkinson" {ajenkinson-at-yahoo.com} ; {Microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 01, 2003 7:18 AM
Gary Whether EDAX is a trademark or not, it is the name of the trading company, and I think you'll find they will be defending it with the necessary, if not overwhelming, force. Interestingly, EDAX do not refer to EDX but use the acronyms EDS, WDS and XMS.
Chris
----- Original Message ----- } From: "Gary Gaugler" {gary-at-gaugler.com} To: "Garber, Charles A." {cgarber-at-2spi.com} Cc: "MSA listserver" {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 02, 2003 4:25 AM
This is an interesting fact. Thanks.
The problem for EDAX, the company, is that they have not defended their alleged trademark. None of their web pages show that EDAX is a registered trademark. Thus, they are in a minority position. Seemingly, a defensive position. But they may not even know this or be aware of this.
If one has a trademark, one must defend it....or risk losing it. The ® makes a big difference.
gary g.
At 06:39 PM 4/1/2003, you wrote: } -- [ From: Garber, Charles A. * EMC.Ver #3.1a ] -- } Attachment: earl.doc Code: 0KTIJ8M \ Created: 04-01-03, 09:36 PM } [124 Kb] } } I think you are incorrect, the term EDAX is a registered trade name with the } US Office of Patents and Trademarks. } Chuck } } } EDAX is a tradename. It is not a trademark. } } It may have been at one time but not now. } } You can check for trademarks at } } } } http://www.uspto.gov and do a search of } } the trademarks database. EDAX returns zero. } } } } Have the procedure revised to kick the A } } out. EDX. That might work. Or, perhaps EDS? } } } } gary g. } } } } } } } } At 06:37 AM 4/1/2003, you wrote: } } } } } Hi } } } Does anyone know when the word "EDAX" was registered } } } as a trademark? } } } The reason I ask is that I have come across procedural } } } documentation that calls for "...analysis by EDAX } } } (Energy Dispersive Analysis of X-Rays)." } } } One lab is now telling us that they cannot comply as } } } the don't have EDAX - theirs is a PGT system ! } } } } } } Ady } } } } } } -------- REPLY, End of original message -------- }
-----Message d'origine----- De : Edward Calomeni [mailto:calomeni-1-at-medctr.osu.edu] Envoyé : mardi 1 avril 2003 15:32 À : Microscopy-at-sparc5.microscopy.com Objet : Re: Third party microscope repairs and service
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Morning Rick,
Why look for another service company, Philips is willing to work on a pay for service rate. Check with them, that way you still can maintain the current service person.
Best of Luck
Ed
Edward Calomeni Director EM Lab Ohio State University - Pathology M018 Starling Loving Hall 320 W. 10th Ave. Columbus, OH 43210-1240 614-293-5580 (office) 614-293-8806 (lab) calomeni-1-at-medctr.osu.edu
} } } Rick Harris {raharris-at-ucdavis.edu} 03/31/03 05:35PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
As part of our restructuring, we will not renew the service contract on our trusty old Philips410LS. Can anyone recommend third party service in Northern California? I understand Ron Veil is no longer in business.
Rick A. Harris, Director Microscopy and Imaging Facility Section of Molecular and Cellular Biology 1241 Life Sciences Addition University of California Davis, CA 530 752 2914 http://katie.ucdavis.edu raharris-at-ucdavis.edu
In a message dated 4/2/03 1:40:57 AM, RVane-at-Evactron.com writes:
} John Russ helped start the confusion in terms. He popularized the } "EDAX" term with the book "Energy Dispersive Analysis of X-rays" } edited by John Russ and published by ASTM in 1971. John then worked } for EDAX Inc. in the 1970s before going to North Caroline State. } } Do you have any more comments on this history, John?
Not really. I've been following the exchange with interest. EDAX Labs was started as a "division" of Nuclear Diodes in 1969, when I left JEOL to head it up. The name was actually proposed by Adrian Moggre, our European salesman. The name started to catch on, and we were a major, even dominant market player (along with PGT and Kevex), so in '71 we changed the name of company. The fact that people confused the name of the company with the name of the technique was mostly a welcome thing as it helped market penetration. EDS and EDX weren't pronouncable, but Edax was. Edax was sold to Philips in'74, and I left in '78 to come here to N. C. State U. (retired in '95). As far as I know there are only two people (Alan Sandborg, who preceded me, and Bob Shen, whom I hired) still at Edax from the "old days." Probably Alan could comment on whether any conscious decision was ever made to not register or defend the name as a trademark.
Dr. Guo Feng Zhang Division of Bioenginerring & Physical Science 301-451-3856 ZhangGuo-at-mail.nih.gov
-----Original Message----- } From: Nestor J. Zaluzec [mailto:zaluzec-at-sparc5.microscopy.com] Sent: Sunday, March 30, 2003 8:18 PM To: NewSub-at-sparc5.microscopy.com
********************************************* This Email contains Important Information about the Microscopy Listserver. Please read it all then SAVE a copy for future reference. - Nestor **************************************** To: NewSub-at-MSA.Microscopy.Com } From: "Nestor J. Zaluzec" {zaluzec-at-microscopy.com}
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I am interested in getting a second hand cryotransfer system for SEM. For example: Fisons LT 7400 Oxford CT 1500
Thank you for suggestions/offers.
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At 10:19 AM 4/2/03 -0600, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
A colleague of mine is planning 3D reconstruction of mouse retina based on EM micrographs. My limited experience was from the manual tracing of scanned micrographs. I wonder any of you had done EM reconstruction lately? What's the recommended internal alignment marker? Any good imaging software to help with the image reconstruction? Any suggestions would be highly appreciated.
I am using Scion Image and a graphics tablet to measure areas of tissues under the microscope. I must use the line tool rather than the automatic function to find the areas, and thus I can get only one area at a time. There are 20 or more areas, sometimes overlapping, to measure per field, which makes the proces difficult if not impossible. Is there a workaround in Scion Image, or perhaps other software compitable with Scion cards, that can measure multiple areas per field? I would appreciate any advice.
If you are absolutely sure that manual tracing is the only way to get the measurements you need, why not put the image into a program like Photoshop (or PAint Shop Pro, etc.) and use it to draw outlines and fill the regions you are interested in with a unique color. then just have Image measure the colored regions.
John Russ =====
In a message dated 4/2/03 6:20:59 PM, gbarclay-at-trinidad.net writes:
} I am using Scion Image and a graphics tablet to measure areas of tissues } under the microscope. I must } use the line tool rather than the automatic function to find the areas, } and thus I can get only one area } at a time. There are 20 or more areas, sometimes overlapping, to measure } per field, which makes the } proces difficult if not impossible. Is there a workaround in Scion Image, } or perhaps other software } compitable with Scion cards, that can measure multiple areas per field? } I would appreciate any advice.
Are you making your paper universally available? If so I would like a copy too.
Cheers,
Jodi Reynolds. ---------- From: Steve Chapman [SMTP:protrain-at-emcourses.com] Sent: Wednesday, 2 April 2003 01:25 To: MSA Cc: ROSSCAC Subject: Quality in Electron Microscopy
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Hi
I have co-written a paper on "Quality in Electron Microscopy" which I would be pleased to send through to you if you are interested?
It deals with test procedures, instrument and staff assessment, quality control in an EM unit etc etc.
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: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, March 31, 2003 1:42 PM Subject: EM Quality Control
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good morning all in the listserver, } I had a question - how does on go about doing quality control on EM - both } TEM and SEM. In other words, other than giving techs unknown samples and } checking the end results - or just looking over their shoulders checking } thicks and thin quality - is there another way? } Thanks in advance, } Connie Cummings } }
Are there are any plant people out there who have used Yariv reagents to disrupt AGPs? I want to know how to prepare the reagents (purcahsed from Biosupplies Australia) so I can put them in a solid agar medium for culturing. Are there are any good references on the technical aspects of using the reagents?
Thanks for your help (I know, not strictly a microscope question!).
Are there are any plant people out there who have used Yariv reagents to disrupt AGPs? I want to know how to prepare the reagents (purcahsed from Biosupplies Australia) so I can put them in a solid agar medium for culturing. Are there are any good references on the technical aspects of using the reagents?
Thanks for your help (I know, not strictly a microscope question!).
Well if I had a $ for every email reply I have had for my offer I would now be planning a l o n g holiday??*!!
One MSA listserver star pointed out to me that to place the paper (now upgraded) on the web would save a good deal of work. So if you need a copy you are welcome to take it from www.emcourses.com.
Thank you all for your interest.
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
I would also appreciate a copy if available. Thank you
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com ----- Forwarded by Robert Fowler/TCU/TDK-US on 04/03/2003 07:15 AM -----
"Reynolds, Jodi JI" To: {microscopy-at-sparc5.microscopy.com} {ReynoldsJ-at-one cc: steel.com} Subject: RE: Quality in Electron Microscopy
04/02/2003 08:44 PM
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Hi Steve,
Are you making your paper universally available? If so I would like a copy too.
Cheers,
Jodi Reynolds. ---------- From: Steve Chapman [SMTP:protrain-at-emcourses.com] Sent: Wednesday, 2 April 2003 01:25 To: MSA Cc: ROSSCAC Subject: Quality in Electron Microscopy
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html -----------------------------------------------------------------------.
Hi
I have co-written a paper on "Quality in Electron Microscopy" which I would be pleased to send through to you if you are interested?
It deals with test procedures, instrument and staff assessment, quality control in an EM unit etc etc.
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: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, March 31, 2003 1:42 PM Subject: EM Quality Control
} } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Good morning all in the listserver, } I had a question - how does on go about doing quality control on EM - both } TEM and SEM. In other words, other than giving techs unknown samples and } checking the end results - or just looking over their shoulders checking } thicks and thin quality - is there another way? } Thanks in advance, } Connie Cummings } }
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-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
When I have been asked in the past to comment on this topic, as a speaker, I have first asked the audience what their definition of quality was and it has always been an amazement to me how passionately individuals can become about "their" definition of quality.
I have always thought that quality meant "meeting or exceeding the customer's expectations". At least that has been our overall guiding philosophy in the management of both our own in-house electron microscopy services laboratory as well as the manufacturing and distribution of our SPI Supplies products.
Speaking specifically of the management of the EM laboratory part of our business, this means that in a sense, each customer could have their own perception of what quality means to them, and we have to understand that and we have to give them what they want in terms of expectations.
Some customers want the fastest possible turnaround time, yet others might want the highest possible resolution in an image. Others are more interested in contrast than necessarily resolution. Others are more interested in the detail and explanation in our final report. Some want the technician actually running the microscope to be readily available by telephone. Yet others, for example, someone measuring the size of latex particles, the correct magnification calibration of the microscopes is the single most important expectation.
And not to be forgotten is that customer who has a problem to solve and has the expectation that we will have the staff that has the ability to technically understand the problem and will take the time to sort things out to the degree that the right control and exemplar samples will submitted and the work will be done in a way that results will be generated to solve the problem.
The list becomes endless.
In order to enhance the chances that they will be operating at the expectation of their customers, there is needed a quality "process", a method by which there can be measured the degree to which the expectations of the customers are indeed being met. Some form of a total quality management system is needed. We ourselves have found that when a job had to be done over again, since the "cost of rework" in an EM lab is so high, it is always helpful to understand what went wrong and why it had to be done over again so that the management can learn from that experience and take steps to keep that kind of problem from happening in the future and thereby lowering the costs of rework and the overall costs of the laboratory's operation overall.
This kind of quality process is also an integral part of any credible laboratory accreditation program and the need to comply is what we needed to be the "stick" that brought about the needed mind set change. So we ourselves happen to see that whatever money we pay for the inspection to be accredited to the standard of ISO 17025, we get back in interest, so to speak, because of the way it has helped us to reduce the number of times a job has to be done over again and therefore also our cost of "rework".
So in the end, for an EM lab to operate with "high quality", it is a matter of whether it is being operated at a level that is either at or greater than the expectations of their customers and for that to happen, there has to be a constant process of feedback in terms of how close that laboratory is coming to meeting the expectations.
Chuck
PS: Remember that we are striving to be 100% paperless, therefore there are no paper copies kept of this correspondence. Please be sure to always reply by way of "reply" on your software so that the entire string of correspondence can be kept in one place. ============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id IAA08820 for dist-Microscopy; Thu, 3 Apr 2003 08:12:47 -0600 (CST) Received: from njz_spm_filter (sparc5 [206.69.208.10]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with SMTP id IAA08816 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Thu, 3 Apr 2003 08:12:16 -0600 (CST) Received: from PHSEXCHICI2.Partners.org (phsexchici2.partners.org [170.223.254.21]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id IAA08809 for {microscopy-at-sparc5.microscopy.com} ; Thu, 3 Apr 2003 08:12:04 -0600 (CST) Received: by phsexchici2.partners.org with Internet Mail Service (5.5.2655.55) id {HTAY6GV1} ; Thu, 3 Apr 2003 09:06:16 -0500 Message-ID: {877B1A0F53E2D411B09400508BF93B9F04FEFEEB-at-phsexch19.mgh.harvard.edu} {microscopy-at-sparc5.microscopy.com}
Steve,
I echo Jodi's response, I too would like a copy of your paper!
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
} -----Original Message----- } From: Reynolds, Jodi JI [SMTP:ReynoldsJ-at-onesteel.com] } Sent: Wednesday, April 02, 2003 8:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: RE: Quality in Electron Microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Steve, } } Are you making your paper universally available? If so I would like a } copy too. } } Cheers, } } Jodi Reynolds. } ---------- } From: Steve Chapman [SMTP:protrain-at-emcourses.com] } Sent: Wednesday, 2 April 2003 01:25 } To: MSA } Cc: ROSSCAC } Subject: Quality in Electron Microscopy } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } Hi } } I have co-written a paper on "Quality in Electron Microscopy" which } I would } be pleased to send through to you if you are interested? } } It deals with test procedures, instrument and staff assessment, } quality } control in an EM unit etc etc. } } 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: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, March 31, 2003 1:42 PM } Subject: EM Quality Control } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Good morning all in the listserver, } } I had a question - how does on go about doing quality control on } EM - both } } TEM and SEM. In other words, other than giving techs unknown } samples and } } checking the end results - or just looking over their shoulders } checking } } thicks and thin quality - is there another way? } } Thanks in advance, } } Connie Cummings } } } } } } } }
I would also like to a copy if available, thanks a lot
Jinguo Wang
Jinguo Wang, Ph.D The Pennsylvania State University Materials Research Institute 194 Materials Research Institute Building University Park, PA 16802 Tel: (814) 865-9285 Fax: (814) 863-8561, (814) 863-0637 email: jqw11-at-psu.edu At 07:16 AM 4/3/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Z slice depth in the Zeiss LSM 510 software is limited to the working distance of the objective you are using. The objective you describe I believe has a working distance of only 90 ums. That means if you are asking it to image deeper into a sample than that it will not do it because it is impossible to image beyond its working distance Check its specs.
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University 609-258-5432
Visit us at http://www.molbio.princeton.edu/facility/confocal/index.html
All Many thanks for all the useful information. I think the best think to do is to remove the offending "A" from "EDAX" as an 'Editorial Change'in our procedure. That should keep everyone happy :-)
Regards Ady
__________________________________________________ Do you Yahoo!? Yahoo! Tax Center - File online, calculators, forms, and more http://tax.yahoo.com
The Z slice depth in the Zeiss LSM 510 software is limited to the working distance of the objective you are using. The objective you describe I believe has a working distance of only 90 ums. That means if you are asking it to image deeper into a sample than that it will not do it because it is impossible to image beyond its working distance Check its specs.
Joe Goodhouse Confocal / EM Core Laboratory Department of Molecular Biology Princeton University 609-258-5432
Visit us at http://www.molbio.princeton.edu/facility/confocal/index.html
Marc Helvey VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 FAX: (408) 428-9555 E-Mail: marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} Internet: http://www.vlsistandards.com {http://www.vlsistandards.com/}
-----Original Message----- } From: Sherwood, Margaret [mailto:MSHERWOOD-at-PARTNERS.ORG] Sent: Thursday, April 03, 2003 6:06 AM To: 'protrain-at-emcourses.com' Cc: 'microscopy-at-sparc5.microscopy.com'
Steve,
I echo Jodi's response, I too would like a copy of your paper!
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
} -----Original Message----- } From: Reynolds, Jodi JI [SMTP:ReynoldsJ-at-onesteel.com] } Sent: Wednesday, April 02, 2003 8:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: RE: Quality in Electron Microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Steve, } } Are you making your paper universally available? If so I would like a } copy too. } } Cheers, } } Jodi Reynolds. } ---------- } From: Steve Chapman [SMTP:protrain-at-emcourses.com] } Sent: Wednesday, 2 April 2003 01:25 } To: MSA } Cc: ROSSCAC } Subject: Quality in Electron Microscopy } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } Hi } } I have co-written a paper on "Quality in Electron Microscopy" which } I would } be pleased to send through to you if you are interested? } } It deals with test procedures, instrument and staff assessment, } quality } control in an EM unit etc etc. } } 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: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, March 31, 2003 1:42 PM } Subject: EM Quality Control } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Good morning all in the listserver, } } I had a question - how does on go about doing quality control on } EM - both } } TEM and SEM. In other words, other than giving techs unknown } samples and } } checking the end results - or just looking over their shoulders } checking } } thicks and thin quality - is there another way? } } Thanks in advance, } } Connie Cummings } } } } } } } }
Quality is what meets the requirements of the job. Anything less has to be done over and anything more is a waste of time and money. Of course you have to aim a little higher than just good enough to do the job to eliminate the ones that fall short. But far too much money is spent on quality that isn't needed for the results desired.
How many times have you seen results to three decimal places based on measurement taken with instruments that are accurate to one decimal place. Computers have only made this worse. When we used slide rules we at least realized that the results were low resolution.
Realistically some one doing work for the public has to do higher quality than the customer would be satisfied with if he was doing it himself but when setting in house quality standards make them only as good as they need to be plus a reasonable safety factor.
Making every job a master piece looks great but it is terrible waste of time and money.
Gordon Couger Stillwater, OK www.couger.com/gcouger
----- Original Message ----- } From: "Garber, Charles A." {cgarber-at-2spi.com} : When I have been asked in the past to comment on this topic, as a speaker, I : have first asked the audience what their definition of quality was and it : has always been an amazement to me how passionately individuals can become : about "their" definition of quality. : : I have always thought that quality meant "meeting or exceeding the : customer's expectations". At least that has been our overall guiding : philosophy in the management of both our own in-house electron microscopy : services laboratory as well as the manufacturing and distribution of our SPI : Supplies products. : : Speaking specifically of the management of the EM laboratory part of our : business, this means that in a sense, each customer could have their own : perception of what quality means to them, and we have to understand that and : we have to give them what they want in terms of expectations. : : Some customers want the fastest possible turnaround time, yet others might : want the highest possible resolution in an image. Others are more interested : in contrast than necessarily resolution. Others are more interested in the : detail and explanation in our final report. Some want the technician : actually running the microscope to be readily available by telephone. Yet : others, for example, someone measuring the size of latex particles, the : correct magnification calibration of the microscopes is the single most : important expectation. : : And not to be forgotten is that customer who has a problem to solve and has : the expectation that we will have the staff that has the ability to : technically understand the problem and will take the time to sort things out : to the degree that the right control and exemplar samples will submitted and : the work will be done in a way that results will be generated to solve the : problem. : : The list becomes endless. : : In order to enhance the chances that they will be operating at the : expectation of their customers, there is needed a quality "process", a : method by which there can be measured the degree to which the expectations : of the customers are indeed being met. Some form of a total quality : management system is needed. We ourselves have found that when a job had to : be done over again, since the "cost of rework" in an EM lab is so high, it : is always helpful to understand what went wrong and why it had to be done : over again so that the management can learn from that experience and take : steps to keep that kind of problem from happening in the future and thereby : lowering the costs of rework and the overall costs of the laboratory's : operation overall. : : This kind of quality process is also an integral part of any credible : laboratory accreditation program and the need to comply is what we needed to : be the "stick" that brought about the needed mind set change. So we : ourselves happen to see that whatever money we pay for the inspection to be : accredited to the standard of ISO 17025, we get back in interest, so to : speak, because of the way it has helped us to reduce the number of times a : job has to be done over again and therefore also our cost of "rework". : : So in the end, for an EM lab to operate with "high quality", it is a matter : of whether it is being operated at a level that is either at or greater than : the expectations of their customers and for that to happen, there has to be : a constant process of feedback in terms of how close that laboratory is : coming to meeting the expectations. : : Chuck : : PS: Remember that we are striving to be 100% paperless, therefore there : are no paper copies kept of this correspondence. Please be sure to always : reply by way of "reply" on your software so that the entire string of : correspondence can be kept in one place. : ============================================ : : Charles A. Garber, Ph. D. Ph: 1-610-436-5400 : President 1-800-2424-SPI : SPI SUPPLIES FAX: 1-610-436-5755 : PO BOX 656 e-mail:cgarber-at-2spi.com : West Chester, PA 19381-0656 USA : Cust.Service: spi2spi-at-2spi.com : : Look for us! : ######################## : WWW: http://www.2spi.com : ######################## : ============================================ : :
I would also appreciate a copy if available. Thank you
dr. Pogány Lajos Senior Research Fellow, Metals Research Department, Research Institute for Solid State Physics and Optics, Hungarian Academy of Sciences Office Address: H-1121 Budapest, Konkoly-Thege ut 29-33 Letters: H-1525 Budapest, P.O.B. 49, Hungary Phone: (00)-36-1-392-2222/17-25; Fax: (00)-36-1-392-2215 e-mail:pogany-at-szfki.hu homepage: http://www.szfki.hu/~pogany/
The University of Texas Health Science Center at San Antonio will host a Symposium and Workshop sponsored by Hamamatsu Photonics KK on
Fluorescence Lifetime Imaging and Spectral Imaging
Take advantage of a unique opportunity to get Hands-On experience with commercially available Spectral Imaigng and Fluorescence Lifetime Imaging Microscopy systems.
Workshop Application Deadline: April 7, 2003 June 8-10, 2003 UT Health Science Center San Antonio, TX Tuition: $700 (includes room and board) 20 student limit/4 scholarships available
Prior to the Workshop a Symposium on Spectral Imaging and Fluorescence Lifetime Imaging Microscopy will be held at the Gunter Hotel. Separate regisistration for the symposium is required.
Symposium Registration Deadline: May 1, 2003 June 6-7, 2003 The Sheraton Gunter Hotel 205 E. Houston St. San Antonio, TX Student: $200 ($250 after May 1) Professional: $250 ($300 after May 1)
Academic Participants: Philippe Bastiaens (GER)*Robert Clegg (USA)*Mary Dickinson (USA)*Paul French (UK) Hans Gerritsen (Neth)*Brian Herman (USA)*Ammasi Periasamy (USA) Herbert Schneckenburger (GER)*Ton Visser (Neth)*Timo Zimmermann (GER)
-----Message d'origine----- De : Helvey, Marc [mailto:Marc.Helvey-at-vlsistd.com] Envoyé : vendredi 4 avril 2003 02:09 À : 'Sherwood, Margaret '; 'protrain-at-emcourses.com' Cc : 'microscopy-at-sparc5.microscopy.com' Objet : RE: Quality in Electron Microscopy
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In case someone hasn't already posted this:
http://www.emcourses.com/quality.htm
Marc Helvey VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 FAX: (408) 428-9555 E-Mail: marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} Internet: http://www.vlsistandards.com {http://www.vlsistandards.com/}
-----Original Message----- } From: Sherwood, Margaret [mailto:MSHERWOOD-at-PARTNERS.ORG] Sent: Thursday, April 03, 2003 6:06 AM To: 'protrain-at-emcourses.com' Cc: 'microscopy-at-sparc5.microscopy.com'
Steve,
I echo Jodi's response, I too would like a copy of your paper!
Peggy
Peggy Sherwood Lab Associate, Photopathology Wellman Laboratories of Photomedicine (W224) Massachusetts General Hospital 55 Fruit Street Boston, MA 02114 617-724-4839 (voice mail) 617-726-6983 (lab) 617-726-3192 (fax) msherwood-at-partners.org
} -----Original Message----- } From: Reynolds, Jodi JI [SMTP:ReynoldsJ-at-onesteel.com] } Sent: Wednesday, April 02, 2003 8:45 PM } To: microscopy-at-sparc5.microscopy.com } Subject: RE: Quality in Electron Microscopy } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi Steve, } } Are you making your paper universally available? If so I would like a } copy too. } } Cheers, } } Jodi Reynolds. } ---------- } From: Steve Chapman [SMTP:protrain-at-emcourses.com] } Sent: Wednesday, 2 April 2003 01:25 } To: MSA } Cc: ROSSCAC } Subject: Quality in Electron Microscopy } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } Hi } } I have co-written a paper on "Quality in Electron Microscopy" which } I would } be pleased to send through to you if you are interested? } } It deals with test procedures, instrument and staff assessment, } quality } control in an EM unit etc etc. } } 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: {"ROSSCAC-at-aol.com"-at-sparc5.microscopy.com} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, March 31, 2003 1:42 PM } Subject: EM Quality Control } } } } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of } America } } To Subscribe/Unsubscribe -- Send Email to } ListServer-at-MSA.Microscopy.Com } } On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } } -----------------------------------------------------------------------. } } } } } } Good morning all in the listserver, } } I had a question - how does on go about doing quality control on } EM - both } } TEM and SEM. In other words, other than giving techs unknown } samples and } } checking the end results - or just looking over their shoulders } checking } } thicks and thin quality - is there another way? } } Thanks in advance, } } Connie Cummings } } } } } } } }
Eastman Chemical Company has a position in its corporate research and development labs for a microscopist. Extensive experience with both optical and electron microscopy is required and experience in AFM, computer programming, and/or particle size analysis would be a strong plus. Knowledge of polymer morphology and/or the morphology of coatings and inks is also highly desirable. A PhD is preferred. The laboratory is well equipped with several optical microscopes, an SEM, a TEM, an AFM, and a particle size analyzer. The successful candidate will be working with this equipment and other scientists and engineers at Eastman to develop new/improved polymers and chemicals and to help solve manufacturing problems. Candidates must be highly motivated, have good communication skills and be able to work with teams.
Eastman Chemical Company, a Fortune 500 company, is a major manufacturer of plastics, fibers, and specialty organic chemicals. This position is at the Kingsport, TN site. Kingsport is located in northeast Tennessee in the foothills of the Smoky Mountains. For great information on the Tri-Cities area check www.tricities.net {http://www.tricities.net} and www.johnsoncitytn.com {http://www.johnsoncitytn.com}
Eastman is an equal opportunity/affirmative action employer.
To submit your CV/Resume: www.eastman.com {http://www.eastman.com} , go to Employment, Positions Available and then click Submit Your Resume Online. Reference code ECO/1064HS.
Louis T. Germinario (Lou) Physical Chemistry Research Laboratory Eastman Chemical Company Lincoln Street, B-150B P.O. Box 1972 Kingsport, TN 37662-5150 (423) 229-4047 (423) 229-4558 (Fax) {mailto:germ-at-eastman.com}
The New England Society for Microscopy (NESM) and the Connecticut Microscopy Society (CMS) are pleased to announce the 20th. Annual Woods Hole Spring Symposium on Microscopy, to be held at the Marine Biological Laboratory, Woods Hole, Massachusetts, May 2-3, 2003.
Three scientific sessions are planned, together with a banquet and after-dinner talk, and an exhibition featuring the sposoring societies' sustaining members. A poster session and photomicrograph display will also be featured. Full details, including program information and registration details, are available in the NESM April newsletter, available online at:
All individuals interested in microscopy are invited to participate. Special registration rates are available for student members of the sponsoring societies. Corporate entities interested in sustaining membership are cordially invited to enquire for details as indicated in the Web pages.
The Electron Microscope Facility at the University of New Hampshire is in the process of buying a 120kV TEM. We are a core facility for the University. We are currently looking at Leo's 912AB Omega EFTEM, JEOL's JEM 1230 (HC) and FEI's Tecnai G2 12 Twin.
Do any list-server members have experience with any of these microscopes? Our applications tend to be predominantly biological although we do have some polymer and materials users/researchers.
Any responses would be helpful and can be e-mailed or telephoned directly to me.
Thank you very much.
Nancy Cherim Electron Microscope Facility University of New Hampshire Kendall Hall Room 6 Durham, NH 03824
The next meeting of the Metropolitan Microscopy Society will be held on April 9th, 2003 at the EDAX facility in Mahwah, NJ. EDAX has graciously agreed to host our meeting and to provide complimentary coffee and lunch to all the attendees.
The forthcoming meeting comprises five presentations offering a blend of microscopic, spectroscopic and image processing applications. Mike Marko will describe the cutting edge methods for tomographic reconstruction of biological organelles. Paul Kotula and Tom Hancewicz will discuss methods to extract maximum value from the data by analyzing spectral images. Matt Libera will illustrate ways to form novel surface patterns on polymeric materials, using electrons, for controlling biological activity and Del Redfern will highlight the advances in high-resolution imaging in projection x-ray microscopy.
We invite all members to attend and to inform their colleagues and fellow workers who may also wish to attend the meeting. This will not only offer an opportunity to listen to some of the experts in their respective fields but also a venue to meet colleagues and exchange ideas as well as to discuss some of the new items that are in the pipeline for this year.
It is important that members should pre-register as it will help us plan for lunches. The pre-registration deadline is April 4 and can be accomplished electronically. Please respond via email or fax to Evan Slow directly. For all attendees, the meeting fee, which includes lunch, will be $20.00.
For any additional information about the meeting, please contact any of the officers. We hope to see you on April 9th at the meeting in Mahwah!
Place: EDAX Inc, 91 McKee Drive, Mahwah, NJ 07430-2120. (201) 529-4880 Mapquest: http://www.mapquest.com/maps/map.adp?country=US&addtohistory=&address=18+mckee+drive&city=mahwah&state=nj&zipcode=&homesubmit=Get+Map · 9:30 - 9:55 Registration and Coffee· 9:55 - 10:00 Introduction Manoj Misra, Unilever Research and Development,Edgewater, NJ· 10:00 - 10:45 Electron tomography: techniques and applications Mike Marko, Wadsworth Research Center, Albany,NY· 10:45 - 11:30 Current Methods in Multivariate Spectroscopic ImageAnalysis Tom Hancewicz, Unilever Research andDevelopment, Edgewater, NJ· 11:30 - 12:15 Electron-beam surface-patterned polymers Matt Libera, Stevens Institute of Technology,Hoboken, NJ· 12:15 - 1:00 Lunch (included with registration ? pleasepre-register!)· 1:00 - 1:30 EDAX Tour· 1:30 - 2:15 Spectral Imaging: The Next Step in Microanalysis Paul Kotula, MAS Sponsored Tour Lecturer,Sandia National Lab, Albuquerque, NM · 2:15- 3:00 Projection X-ray microscopy in the SEM Del Redfern, EDAX, Mahwah, NJ
by sparc5.microscopy.com (8.9.3+Sun/8.9.3) id QAA03513 for dist-Microscopy; Mon, 7 Apr 2003 16:05:01 -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 QAA03506 for "MicroscopyFilteredEmail3-at-msa.microscopy.com"; Mon, 7 Apr 2003 16:04:30 -0500 (CDT) Received: from postmark.nist.gov (pushme.nist.gov [129.6.16.92]) by sparc5.microscopy.com (8.9.3+Sun/8.9.3) with ESMTP id QAA03499 for {Microscopy-at-sparc5.Microscopy.Com} ; Mon, 7 Apr 2003 16:04:18 -0500 (CDT) Received: from SCOTT-OPTI.nist.gov (mag216.nist.gov [129.6.98.216]) by postmark.nist.gov (8.12.5/8.12.5) with ESMTP id h37KwKbe023464 for {Microscopy-at-MSA.Microscopy.Com} ; Mon, 7 Apr 2003 16:58:20 -0400 (EDT) Message-Id: {5.1.1.5.2.20030407163719.03b66118-at-postmark.nist.gov} X-Sender: js0b-at-postmark.nist.gov X-Mailer: QUALCOMM Windows Eudora Version 5.1.1
Meeting Announcement
REGISTRATION DEADLINE: Friday April 11, 2003
A 4-day MAS Special Topics Workshop on spectrum-imaging and multispectral & hyperspectral data analysis will be held at NIST in Gaithersburg, Maryland from April 28th through May 1st, 2003.
The event will cover forms of hyperspectral data relevant to microbeam analysis, including STEM-based EELS and XEDS, energy-filtered TEM imaging, XEDS in the SEM, spectrum imaging in mass spectrometry, XPS, and optical spectroscopies such as IR and cathodoluminescence.
While the original registration deadline has passed and the technical program is full, a limited number of attendance slots are available. There will also be room for additional posters if you wish to contribute or present scientific material or discuss your ideas on hyperspectral analysis.
If you would like to register for the workshop, please send an email to {mailto:johnhenry.scott-at-nist.gov} including:
Name Affiliation Telephone Number Country of Citizenship which days you will likely attend
I will reply as soon as possible to confirm your registration if slots are still available.
Details about the meeting can be found in the
Workshop Announcement PDF http://www.microprobe.org/masjh/TCs/2003-Hyperspectral/2003-Workshop-Announcement.pdf
or at the Workshop Announcement web page http://www.microprobe.org/masjh/TCs/2003-Hyperspectral/2003-Workshop-Announcement.html
Some Confirmed Invited Speakers:
· Ian Anderson, Oak Ridge National Laboratory · Jim Bentley, Oak Ridge National Laboratory · John Friel, Princeton Gamma-Tech, Inc. · Gerald Kothleitner, FELMI TU-Graz · Paul Kotula, Sandia National Laboratories · Richard Leapman, National Institutes of Health · Paul Mainwaring, Gatan, Inc. · Robert Martin, University of Strathclyde · Chris Michaels, NIST · Larry Nittler, Carnegie Institution of Washington · Michaeleen Pacholski, Rohm and Haas Company · Diane Peebles, Sandia National Laboratories · Peter Statham, Oxford Instruments · Masashi Watanabe, Lehigh University · Norman Wright, Digilab, LLC
Thanks to the generosity of NIST, MAS, and our corporate sponsors attending the workshop, there is no registration fee.
Sponsors
4Pi Analysis, Inc. Advanced Microbeam, Inc. EDAX International Eigenvector Research, Inc. EmiSpec Systems, Inc. FEI Company Gatan, Inc. JEOL USA, Inc. Oxford Instruments Princeton Gamma-Tech, Inc. Research Systems, Inc. Thermo NORAN Microbeam Analysis Society
I hope to see you in Gaithersburg at the Workshop,
-- John Henry
John Henry J. Scott Bldg 222/Rm A113 NIST Microanalysis Research Group 100 Bureau Drive Stop 8371 (301) 975-4981; (301) 417-1321 FAX Gaithersburg, MD 20899-8371
I hope you will plan to join us this Thursday and Friday at the "Art of the Science Image" conference at the University of Tulsa. We're looking
forward to an exciting two days of lectures, workshops, and discussions about scientific imaging with speakers of great expertise and national reputation. For more detail about the conference, accomodations, and travel directions see the Oklahoma Microscopy Society (OMS) web page below.
We’ll explore the connection between human vision and imaging, delve into spectral imaging and x-ray mapping, tackle depth of field issues, and have the enviable opportunity of a half-day workshop on image analysis with renowned expert Dr. John Russ.
Our not-to-be-missed dinner event will feature a keynote address by David Scharf and a showing of the movie for which his unique blend of science and art received an Emmy award. Plan to enjoy continued conversation with the speakers and your fellow microscopists even after the event by staying at the conference hotel.
We’ll celebrate aesthetics with a workshop on how to create stunning polarized light images, a lecture on the basics of image composition, and a fascinating talk from author and art historian Lynn Gamwell on connections past and present between microscopy and art. An exhibit of scientific photography from the Rochester Institute of Technology will be on display along with works by David Scharf.
Corporate exhibitors and underwriters with whom you'll the the opportunity to meet and discuss equipment are: Gatan, MicroStar, IXRF, Olympus, Princeton Gamma Tech, Thermo NORAN, Meyer Instruments (Leica Microsytems
and Image-Pro), JEOL, Nikon, Atomic Spectroscopy Instruments, BioRad, and Hitachi.
-- ================================================================== Greg Strout Electron Microscopist, University of Oklahoma WWW Virtual Library for Microscopy: http://www.ou.edu/research/electron/www-vl/ e-mail: gstrout-at-ou.edu (405)325-4391 Opinions expressed herein are mine and not necessarily those of the University of Oklahoma ==================================================================
I'm making up a .28 M stock solution for veronal acetate buffer. 1.47g barbituric acid, 0.97g sodium acetate trihydrate, carbon dioxide free water to 50 ml. I can't get this to dissolve. Does anyone know how to get this to go into solution? Jeannie Selker {jselker-at-molbio.uoregon.edu}
} I'm making up a .28 M stock solution for veronal acetate buffer. } 1.47g barbituric acid, 0.97g sodium acetate trihydrate, carbon } dioxide free water to 50 ml. I can't get this to dissolve. Does } anyone know how to get this to go into solution? Jeannie Selker } {jselker-at-molbio.uoregon.edu}
Are you sure you want barbituric acid as it is notoriously hard to dissolve? Most people use sodium barbitol (or veronal as it is sometimes called). Also it sounds too concentrated.
The formula that I use is: 2.96 g sodium barbitol, 1.94 g sodium acetate (hydrated) made up to 100 ml. You need to stir this vigorously (and even you can heat it to 70-80 deg. C with stirring). So, use a heated, magnetic stirrer that is vigorously stirring the solution. Be patient as it may take a long time to dissolve (sometimes over an hour).
The buffer is then completed by mixing: 12.5 ml of your fixative (2% osmium for example, in ddwater), 5.0 ml of the above veronal solution, 2.5 ml of ddwater, and 0.1N HCl to adjust to the final pH (around 5 ml for a pH of 7.4).
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 Fax: 618-453-2665 Email: bozzola-at-siu.edu ##############################################################
We have the fei Technai 12 TEM. The scope is really nice. We have a twin lens and not the biotwin which suits us fine since we are suppose to serve all disciplines from one scope. The scope is great with relative easy alignment for biological science. "Auto stitch" is a very useful function (only free if negotiated during purchase!) stitching more than one pic eg. 3X3 together to increase your resolution. It works nice for biological sciences but the maximum magnification at which it works nice is a bit low for other sciences. The stigmators are the difficult part in the alignment procedure (I might be the only user struggling with it!). The problems we had is with the third party peripherals. Which ever scope you buy just make sure that it all is working properly and that the scope will do what you like in the future. Service and the service provider to me is part of the scope specs since it is horrible to be left without good or slow support. These are my comments. Hope it is of some help.
-----Original Message----- } From: Nancy Cherim [mailto:nac-at-cisunix.unh.edu] Sent: Monday, April 07, 2003 8:46 PM To: Microscopy-at-sparc5.microscopy.com
The Electron Microscope Facility at the University of New Hampshire is in the process of buying a 120kV TEM. We are a core facility for the University. We are currently looking at Leo's 912AB Omega EFTEM, JEOL's JEM 1230 (HC) and FEI's Tecnai G2 12 Twin.
Do any list-server members have experience with any of these microscopes? Our applications tend to be predominantly biological although we do have some polymer and materials users/researchers.
Any responses would be helpful and can be e-mailed or telephoned directly to me.
Thank you very much.
Nancy Cherim Electron Microscope Facility University of New Hampshire Kendall Hall Room 6 Durham, NH 03824
Further, with regard to barbituric acid, I just checked the Merck Index and they say "Difficultly sol in cold water; freely sol in hot water, in dil acids."
Regarding sodium barbitol: "One gram dissolves in 5 ml water, 2.5 ml boiling water, 400 ml alc. Aq soln is alkaline to litmus and phenophthalein. Ph of 0.1 molar aq soln, 9.4."
So, hot water is the way to go.
############################################################## 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 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
Hi, I was wondering if anybody can tell me where to buy a centrifugation adapter. I have dilute virus samples that I would like to pellet directly onto a grid.
I'm not a member of this group. Therefore, I would appreciate it if you could send your response directly to my e-mail address in addition to posting it.
Thank a lot.
Thomas
thomas.weber-at-mssm.edu -- Dr. Thomas Weber Assistant Professor The Carl C. Icahn Center for Gene Therapy and Molecular Medicine Box 1496 Mount Sinai School of Medicine 1425 Madison Avenue New York, NY 10029-6574 United States of America
Microscopy, your immediate help is needed. We are a .com corporation that is growing fast (over 1000% per year). We simply cannot keep up with demand. We are searching for motivated individuals who are looking to earn a substantial income working at home.
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I have a researcher trying to get decent images of mouse egg microvilli. So far no luck. Early SEM examples had an obvious layer of gunk (PVA?) on the the egg. Recent trials show collapsed or adherent microvilli. I have indicated below the articles they were referencing for their prep. Currently, they are fixing, removing zona, dehydrating in EtOH, then CPD. Any suggestions would be appreciated.
For SEM:
-Phillips, D.M. and Shalgi, R. Surface Architecture of the Mouse and Hamster Zona Pellucida and Oocyte. 1980; J. of Ultrastructure Research 72, 1-12
-Shalgi, R. and Phillips, D.M. Mechanics of in vitro Fertilization in the Hamster Biology of Reprod. 1980; 23, 433-444
-Fulka Jr, J., Flechon, B. and Flechon, J.E. Fusion of Mammalian oocytes: SEM observations of surface changes. Reprod. Nutr. Dev. 1989; 29, 551-558
For TEM:
1: Albertini DF, Combelles CM, Benecchi E, Carabatsos MJ. Cellular basis for paracrine regulation of ovarian follicle development. Reproduction. 2001 May;121(5):647-53. Review. PMID: 11427152 [PubMed - indexed for MEDLINE]
2: Albertini DF, Rider V. Patterns of intercellular connectivity in the mammalian cumulus-oocyte complex. Microsc Res Tech. 1994 Feb 1;27(2):125-33. PMID: 8123905 [PubMed - indexed for MEDLINE]
3: Ducibella T, Anderson E, Albertini DF, Aalberg J, Rangarajan S. Quantitative studies of changes in cortical granule number and distribution in the mouse oocyte during meiotic maturation. Dev Biol. 1988 Nov;130(1):184-97. PMID: 3141231 [PubMed - indexed for MEDLINE]
I have an Oxford QX2000 free to a good home, donee to pay packing costs and freight.
It was in working order when it was taken offline a few years ago, apart from a seriously drifty XP2 pulse-processor board.
Unit is complete with monitor and keyboard, and has a few options, the exact nature of which I can't remember, but which included the ability to show a BSE image and position the beam from it.
cheers
rtch
Ritchie Sims Phone : 64 9 3737599 ext 7713 Department of Geology Fax : 64 9 3737435 The University of Auckland email : r.sims-at-auckland.ac.nz Private Bag 92019 Auckland New Zealand
first, i have no commercial interest, but i was involved in the early work developing the use of the beckman Airfuge EM-90 rotor. i have used it extensively for quantification of virus, determining species of subviral components produced and their proportions, immunoEM, and about everything else i can think of doing with suspensions of viruses and bacteria. two papers bracketing the work and covering your specific questions are:
Hammond, GW, Hazelton, PR, Chuang, I, and Klisko, B. 1981. Improved Detection of viruses by electron microscopy after direct ultracentrifuge preparation of specimens. J. Clin. Micro., 14:210-221.
Hazelton, PR and Coombs, KM. 1999. The Reovirus Mutant tsA279 L2 Temperature- Sensitive Lesion Is Associated with Production of a Core Particle Deficient in the lambda 2 Core Spike Protein and Minor Core Protein sigma 2. Journal of Virology, 73:2298-2308.
also, i would refer you to the recent review article by hans gelderblom and i, where we discuss a number of concentration techniques.
Hazelton, PR and Gelderblom, HR. 2003. The use of the electron microscope for rapid diagnosis of viral agents in emergent situations. Emerging Infectious Diseases 9:294-303.
if you have any questions, or need any help, do not hesitate to contact me.
paul r. hazelton, PhD electron microscopist, Department of Medical Microbiology University of Manitoba 531 Basic Medical Sciences 730 William Avenue Winnipeg, MB, Canada, R3E 0W3 Clinical Virology consultant, Cadham Provincial Laboratory Winnipeg, Manitoba, Canada telephone 204-789-3313 pager: 204-931-9354 fax: 204-789-3926 e-mail: paul_hazelton-at-umanitiba.ca
We have the remains of two Oxford Instruments AN10000s - somewhat knocked about but still a valuable source of spares - and an entire LEMAS stage/spectrometer control system. The LEMAS was configured for a 4-WD spectrometer microprobe and in good working order when it was taken out last year - again a very useful source of spares for anybody still keeping one of those going.
Cheers,
Stu ---------------------------- Stuart Kearns Department of Earth Sciences University of Bristol Bristol, UK BS8 1RJ Tel: +44 117 954 5435 Fax: +44 117 925 3385 Stuart.Kearns-at-bristol.ac.uk ----------------------------
Is there freeware out there or a plug in to ifranview for batch printing of images 4 images per page of all selected images?
I really love this list. I just want to thank you all for your helpful comments on questions posted here. Thank you Nestor for keeping it up and running.
Stephan H Coetzee Department of Physics EMU Private Bag 0704 Gaborone, Botswana
I am not sure about the plugins, but if you use windows XP, it has "Windows picture and fax viewer" which does the batch printing.
Pavel
----- Original Message ----- } From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} To: "Listserver Microscopy (E-mail)" {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 09, 2003 5:54 AM
Hi everyone,
I have a Hitachi H600 TEM (20 years old). I think the HV cable is dying. My evidence for this is:
1. Unstable voltage readouts. When we set the voltage to, say, 50 Kv, it does not read 50 Kv but reads 75Kv. 2. There is a 'hissing' sound when we try to change from 75Kv to 100Kv. The hissing appears to be coming from behind the gun. 3. Our filaments, when they break, are showing signs of over-heating as evidenced by a round blob of tungsten on each of the broken ends. 4. When I open the gun, it smells.
We need to keep it alive for another 12-18 months by which time we anticipate replacing it.
Are my conclusions re the HV cable correct? Does anyone out there know if it is possible to repair the HV cable and if so, what is involved?
cheers
Sarah Ellis
Manager, Microscopy Imaging and Research Core Facility Peter MacCallum Cancer Centre Locked Bag #1 A'Beckett Street East Melbourne 8006
Sorry to be a bother, but I have a couple of queries.
I have looked in the archives and hope that I haven't missed any relevant info.
Firstly, I'm looking for the name/product no. of a Dow Corning product that can be used for moulding to make replicas for microscopy. In the past I have used simple latex, but heard Dow did something better. I have looked at the D-C web site but couldn't work out exactly what product to buy. I want to use this stuff for cleaning purposes (AFM standards) and also for making actual replicas, so the name of the right casting product would also be very much appreciated.
Secondly, I'm interested in pattern recognition (for AFM, SEM, TEM etc.). Am I right in thinking this should be a relatively simple branch of image analysis? I know that one can use image analysis to do things like count cells on agar plates via a live camera, or from an image. Can it be done for things like recurring patterns in say crystals? Is there anything available out there that will do this and is free or quite cheap (is there any add to Scion Image that will do it without the need for too many macros etc.?)? Finally, is there any web-based resource/text on image analysis that would cover from beginnings up to a reasonable sophistication, or what is/are the best available books at present?
Thank you all for your patience and thanks in advance to anyone who is able to help.
Are there any references or experienced folks out there that are doing immunoEM for TEM and antigen retreival for archived material and stuff soaking in 2.5% glut?
I'm checking into the method and have come up with nothing so far.
Thanks!
Paula :-)
Paula Sicurello George Washington Univ. Medical Center Dept. of Pathology, Ross Hall rm 505 Electron Microscope Lab 2300 Eye St. Washington, DC 20037 202-994-2930 phone 202-994-2518 fax
There are several options, none that I am aware of that are free or shareware.
The one I us is Thumbs Plus from Cerious Software. It costs about $30. Another program is Image AXS, about the same price. These allow variable number of thumbs per page, depending on the size of the thumbs and page size.
If you make thumbnails as a contact page, you can output them to Acrobat Distiller and save as PDF. They are small file sized and high fidelity output. Current Acrobat is 5.0.
gary g.
At 02:54 AM 4/9/2003, you wrote:
} Dear All } } Is there freeware out there or a plug in to ifranview for batch printing of } images 4 images per page of all selected images? } } I really love this list. I just want to thank you all for your helpful } comments on questions posted here. Thank you Nestor for keeping it up and } running. } } Stephan H Coetzee } Department of Physics } EMU } Private Bag 0704 } Gaborone, } Botswana } } Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} } } Telephone: (+267) 355 2462 } Fax: (+267) 3185 097 } Telephone: (+267) 355 0000 Switchboard
Hi listers My compressor on the cooling tower for my JEOL T-220A has failed. Can anyone refer me to a local (stateside) resource for acquiring this compressor. The system I have now is R-12. I would prefer to change over to R-134. Thank you
Robert Fowler Quality Assurance Failure Analysis Technician TDK Components USA, Inc. Multilayer Ceramic Capacitor Division 1 TDK Boulevard Peachtree City GA 30269-2051 Telephone: (770) 631-0410 Ext.315 Fax: (770) 487-1460 email: rfowler-at-tdktca.com www.component.tdk.com
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If you are not the intended recipient, be advised that any use, dissemination, distribution or duplication of this transmission is strictly prohibited. If you received this transmission in error, please notify the sender immediately by electronic reply to this transmission or by phone (847-803-6100). Thank you.
Stephan, You should have a look at XnView ( www.xnview.com ). It doesn't do batch printing, but it will create batch contact sheets that you can then batch print. Its a great all round program for working with piles of images. ACDSee does a good job with batch printing, but it's not free (CAD$ 70).
Cheers
Glenn
Glenn Poirier
Microbeam Specialist Mineralogy and Metallurgical Processing Mining and Mineral Sciences Laboratory, CANMET 555 Booth St Ottawa, ON K1A OG1
Spécialiste en microfaisceau Minéralogie et procédés métallurgiques Laboratoires des mines et des sciences minérales de CANMET 555, rue Booth Ottawa, Ontario K1A 0G1
tel: (613) 947-9833 FAX: (613) 996-9673 -----Original Message----- } From: Coetzee, Mr S. H Physics Science [mailto:COETZEES-at-mopipi.ub.bw] Sent: 09 April, 2003 05:54 AM To: Listserver Microscopy (E-mail)
Dear All
Is there freeware out there or a plug in to ifranview for batch printing of images 4 images per page of all selected images?
I really love this list. I just want to thank you all for your helpful comments on questions posted here. Thank you Nestor for keeping it up and running.
Stephan H Coetzee Department of Physics EMU Private Bag 0704 Gaborone, Botswana
Photoshop 7 (and 5 too, but with fewer options) has a File--} Automate--} Contact Sheet II feature for making contact sheets of variable # of images, names, PPI, etc. which can the be printed. Basically, you chhose the directory of images and the format you want and it automatically makes the sheets. It does not, however, batch save or print them.
At 07:31 AM 4/9/2003 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
____________________________________________________________________________ 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/
I have been using Qimage Pro 2003 that does a great job and has quite a few capabilities. It allows you to print different numbers of images per page, put the file name underneath the image, draw a line around the image, etc, etc. You can check it out and get a trial copy at
www.ddisoftware.com/qimage
it costs about $35
Usual disclaimer, Damian Neuberger Senior Research Scientist Baxter Healthcare Corp.
----- Original Message ----- } From: "Coetzee, Mr S. H Physics Science" {COETZEES-at-mopipi.ub.bw} To: "Listserver Microscopy (E-mail)" {microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 09, 2003 4:54 AM
Rick,
Aside from the fixation and sample prep, what type of SEM are you using. I did some extensive work with mouse peritoneal microvilli using approximately the same general prep methods. I took some very nice images of the microvilli at fairly high magnification using our JEOL Field Emission SEM at a couple of keV and low beam current. Unfortunately, near the end of the study, I got the call for my lung transplant and so couldn't complete the SEM work on the last samples. They sent them to an outside lab and the photos showed the same kind of damage to the microvilli you describe. Seems that the other lab used a standard tungsten filament SEM at 20keV and unknown beam currrent. It looked like beam damage to me. Could this be an issue for mouse egg samples?
Damian Neuberger Senior Research Scientist Baxter Healthcare Corp.
----- Original Message ----- } From: "Rick Harris" {raharris-at-ucdavis.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Tuesday, April 08, 2003 3:48 PM
Dear Stephan,
You wrote:
} Is there freeware out there or a plug in to ifranview for batch printing } of images 4 images per page of all selected images? } For some time we have been using a program called PrintStation which will allow you to have from one to however many images you want per page. It is very easy to set up and all our users love it. At US$19.95 it is also very cheap. You can find it at : http://www.picmeta.com/printstation.htm
David Vowles Electron Microscope Unit Dept of Materials Science and Metallurgy University of Cambridge Pembroke St Cambridge UK CB2 3QZ Tel: +44 1223 334325 Fax: +44 1223 334567 Email: djv23-at-cam.ac.uk
I have tried to maintain a good database of third-party service providers in the past, but recent expansions in this field have probably outdated the resources I currently list. I would appreciate it if anyone out there who provides services for SEM, TEM and x-ray microanalysers would send their particulars (company name, contact name, email, web site, phone number, fax number, address, coverage area, instruments serviced, services offered).
I receive many requests for sales, service, de-installation, installation and training that are outside the area I cover (the great fly-over country of the American Midwest) - from as far away as Pakistan. I always like to provide competitive referrals whenever possible.
I'd be happy to provide compilations to those who express an interest, and frankly, will soon update my website to provide such referrals in a simple format.
Allen R. Sampson Advanced Research Systems 317 North 4th. Street St. Charles, Illinois 60174
I can't say for certain whether it is your cable that's going. But I did have an old Siemens IA cable go on me back in the 1970s. It was quite spectacular because first there was a loud crack and for a little while there seemed to be a bit of a plasma-like glow at the breakdown point.
Cables may differ but I can tell you that ours went quickly giving no warning and the only recourse we had was to look for a second hand or re-conditioned cable, which we managed to source. I think the only form of repair possible then would have been to shorten the cable but the cost of re-connecting to the gun would be as much as a second hand cable and would not be provided by the manufacturer so there was a risk that it might not be reliable. Try asking a friendly e.m. engineer or Hitachi - they can be quite friendly sometimes.
Malcolm Haswell e.m. unit University of Sunderland UK
----- Original Message ----- } From: "Ellis, Sarah" {s.ellis-at-pmci.unimelb.edu.au} (by way of MicroscopyListserver)
Hi Sarah,
We had a HV cable problem with our Hitachi 2500 SEM and we heard a hissing noise whenever we tried to increase magnification and it would go worse and worse each time.We removed the suspect HV cable and it was cracked inside which caused arching when we turned it on.We were not able to repair it but since our SEM has 2 detectors and we only use one of them we just switched the cables and observation detector while we were waiting for a new cable.
Hope this helps
Virginia Soares INESC Microsistemas e Nanotecnologias R.Alves Redol,9 1000-029 Lisboa PORTUGAL Tel:+351 21 3100300 ext 2504 Fax: +351 21 314 58 43 URL: www.inesc-mn.pt
Well it looks as if you may be correct but there are two steps that you need to take.
1. Follow a strict gun cleaning procedure - take out the cable (care heavy) and if you know how remove the gun chamber (also heavy). If not we can still go ahead. Clean the chamber (if it is in place WITH GREAT CARE NOT TO GET THE MEDI NEAR ANY JOINTS as it will be hard to remove) with a metal polish soluble in ammonia solution. Clean this away with 10:1 water plus ammonia solution on a fluff free but absorbent tissue. Do not let the tissue get too wet, the solution must not run off the tissue when you wipe the chamber. Clean the glazed gun ceramic and all its components with metal polish or where possible in an ultrasonic cleaner in 1:3 water plus ammonia solution (strong). Clean each piece in the US cleaner on its own to prevent scratching. Heat the gun chamber and the metal pieces to drive off residual media and check the gun pieces with a 10X hand lens before re assembly. Check all "o" ring surfaces and "o" rings as you re assemble. This may be enough?
2. The "final" test. Switch off the high voltage and clean round the connection which goes into the high voltage tank. Have some clean paper towel ready, the type that will not leave fluff on a surface. Release the bolts and carefully remove the high voltage cable connection from the tank. Cover the hole in the tank with the clean paper after covering the high voltage connection. Switch on the microscope. If the high voltage levels and standing current are back to normal YES you do have a cable problem.
For greater detail have a look at our CD "Monitoring and Maintaining EM".
If the cable is the problem you are able to have it repaired in your own country. Any organisation that works with high voltage (e.g. hospital x-ray sets etc) will be able to attach a new cable to your old cable ends.
Good luck
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: "Ellis, Sarah (by way of MicroscopyListserver)" {s.ellis-at-pmci.unimelb.edu.au} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 09, 2003 1:46 PM
Does anyone know of a current phone number/address for the ETP/SEMRA company, who manufactured the Robinson detectors? My last address for them was Livermore, CA; however, that phone number is not valid anymore. This company may have merged with someone else. Any help would be greatly appreciated. Thank you in advance. Cathy Kelloes
we are trying to make a in-situ heating stage for a vg501 STEM and are unable to get factory drawings of the stage or sample cartridge. we are concerned with the tapered fit that postions the cartridge. any help would be welcome.
Michael T. Marshall Research Engineer University of Illinois Frederick Seitz Materials Research Laboratory 104 South Goodwin Avenue Urbana, IL 61801 Voice: 217/265-5380 Fax: 217/244-2278 marshall-at-mrl.uiuc.edu
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (jimekstrom-at-attbi.com) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Thursday, April 10, 2003 at 08:09:24 ---------------------------------------------------------------------------
Email: jimekstrom-at-attbi.com Name: James Ekstrom
Organization: Phillips Exeter Academy
Education: 9-12th Grade High School
Location: Exeter, NH USA
Question: I am looking for a copy of the manual for a Nikon Inverted - Phase Contrast microscope. (If you have such a manual I can make a copy and send the original back to you.) I do not have the model number, but I can send along a small JPEG image of the scope if you think you might have a manual for this older microscope. Any assistance would be appreciated.
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It sounds to me like you may have a problem with your vacuum system. I'm not really familiar with Hitachi microscopes but on the JEOL microscopes a smell in the gun usually indicated backstreaming oil from the diffusion pump. Poor vacuum combined with oil in the gun chamber would cause discharging in the gun at higher accelerating voltages. That could also explain the hissing (discharging) sound and the problems with filaments.
Bill
Hi everyone,
I have a Hitachi H600 TEM (20 years old). I think the HV cable is dying. My evidence for this is:
1. Unstable voltage readouts. When we set the voltage to, say, 50 Kv, it does not read 50 Kv but reads 75Kv. 2. There is a 'hissing' sound when we try to change from 75Kv to 100Kv. The hissing appears to be coming from behind the gun. 3. Our filaments, when they break, are showing signs of over-heating as evidenced by a round blob of tungsten on each of the broken ends. 4. When I open the gun, it smells.
We need to keep it alive for another 12-18 months by which time we anticipate replacing it.
Are my conclusions re the HV cable correct? Does anyone out there know if it is possible to repair the HV cable and if so, what is involved?
cheers
Sarah Ellis
Manager, Microscopy Imaging and Research Core Facility Peter MacCallum Cancer Centre Locked Bag #1 A'Beckett Street East Melbourne 8006
You can get quite a few varieties of impression materials from any dental supplier. I used to use them, especially the silicone ones, to make replicas. The casting material is epoxy, such as an Epon substitute or one of the resins made by Epotek. Hope this helps.
Lesley Weston.
on 09/04/2003 5:45 AM, Chris Peppiatt (by way of MicroscopyListserver) at chris.peppiatt-at-nuigalway.ie wrote:
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Dear All, } } Sorry to be a bother, but I have a couple of queries. } } I have looked in the archives and hope that I haven't missed any relevant } info. } } Firstly, I'm looking for the name/product no. of a Dow Corning } product that can be used for moulding to make replicas for } microscopy. In the past I have used simple latex, but heard Dow did } something better. I have looked at the D-C web site but couldn't work } out exactly what product to buy. I want to use this stuff for } cleaning purposes (AFM standards) and also for making actual } replicas, so the name of the right casting product would also be very } much appreciated. } } Secondly, I'm interested in pattern recognition (for AFM, SEM, TEM } etc.). Am I right in thinking this should be a relatively simple } branch of image analysis? I know that one can use image analysis to } do things like count cells on agar plates via a live camera, or from } an image. Can it be done for things like recurring patterns in say } crystals? Is there anything available out there that will do this and } is free or quite cheap (is there any add to Scion Image that will do } it without the need for too many macros etc.?)? Finally, is there any } web-based resource/text on image analysis that would cover from } beginnings up to a reasonable sophistication, or what is/are the best } available books at present? } } Thank you all for your patience and thanks in advance to anyone who } is able to help. } } Chris Peppiatt }
} ---------------------------------------------------------- } -------------- The Microscopy ListServer -- Sponsor: The } Microscopy Society of America To Subscribe/Unsubscribe -- } Send Email to ListServer-at-MSA.Microscopy.Com On-Line Help } http://www.msa.microscopy.com/MicroscopyListserver/FAQ.htm } l } ---------------------------------------------------------- } -------------. } } Does anyone know of a current phone number/address for the } ETP/SEMRA company, who manufactured the Robinson } detectors? My last address for them was Livermore, CA; } however, that phone number is not valid anymore. This } company may have merged with someone else. Any help would } be greatly appreciated. Thank you in advance. Cathy } Kelloes
Here's a followup on my not-so-recent query of The List:
} ... bought a suitable video adapter with C mount for my company's } stereomicroscope and then bought a Kodak MDS 100 camera, both on } eBay,
Took two tries. The first firm set impossible conditions and so my money order was returned unclaimed. The second fellow was quite responsive and the camera arrived factory sealed and installed like a dream, thanks to Kodak's software provider and a second CD hastily stuffed inside the box at "the factory."
} ... in my price range (less than $500 in it so far).
.. snippage ...
One question remains:
} Are there any wavelength issues, such as poor image quality due } to infrared seeping through all that glass ?
Several Listers warned me about this, and sure enough, the image is too red (correctable) and rather fuzzy. There is clearly the need for a filter transparent to visible light but opaque to the long- wavelength infrared light that's causing what I presume to be lots of chromatic abberation. What's such a thing called ?
Best regards, George Langford, Sc.D. Principal Consultant Amenex Associates, Inc. amenex-at-amenex.com http://www.amenex.com/
Hello, I must purchase indoor oil mist exhaust filters for six rotary pumps that are attached to EMs, sputter and carbon coaters and a freeze dryer. Can someone please suggest the most practical, easy to maintain economical units from their experience? Thank you. Regards, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
Here are the contact details of Dr Viv Robinson, and the company details.
Dr. Vivian Robinson ETP Semra Pty Ltd
244 Canterbury Rd Canterbury, NSW 2193
Ph: +61 (2) 9718 1444 Fax: +61 (2) 9718 8222
E-mail: viv-at-etpsemra.com.au
Regards George
----Original Message----- } From: Kelloes, Cathy L [mailto:KELLOECL-at-bp.com] Sent: Thursday, 10 April 2003 9:24 PM To: MSA (E-mail)
Does anyone know of a current phone number/address for the ETP/SEMRA company, who manufactured the Robinson detectors? My last address for them was Livermore, CA; however, that phone number is not valid anymore. This company may have merged with someone else. Any help would be greatly appreciated. Thank you in advance. Cathy Kelloes
Marc Helvey Strategic Accounts Manager VLSI Standards, Inc. 3087 North First Street San Jose, CA 95134-2006 Phone: (408) 428-1800, ext. 108 E-Mail: marc.helvey-at-vlsistd.com {mailto:marc.helvey-at-vlsistd.com} Internet: http://www.vlsistandards.com {http://www.vlsistandards.com/}
-----Original Message----- } From: Kelloes, Cathy L [mailto:KELLOECL-at-bp.com] Sent: Thursday, April 10, 2003 4:24 AM To: MSA (E-mail)
Does anyone know of a current phone number/address for the ETP/SEMRA company, who manufactured the Robinson detectors? My last address for them was Livermore, CA; however, that phone number is not valid anymore. This company may have merged with someone else. Any help would be greatly appreciated. Thank you in advance. Cathy Kelloes
A daylight LBD filter will be the first one needed followed by an IR filter. Each goes in different places, depending on your particular scope. The IR filter is usually at the lamp house. The LBD is at the transport lens or in the vertical illuminator.
gary g.
At 02:09 PM 4/10/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Assuming no toxic exhaust is present, and pumps are not huge, I would use the following:
Some of the best filters are made by Edwards Vacuum (now BOC Edwards). Less expensive OK filters made by Alcatel. Both have easily replaceable cartridges.
See both at these sites, or look up direct links (google will do) http://www.kurtlesker.com/ http://www.duniway.com/ Both these sites have variety of filters.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: "Jim Romanow" {bsgphy3-at-uconnvm.uconn.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, April 10, 2003 6:43 PM
Thanks to all of you who responded in reference to my request on contacts for the address of ETP-USA.
Jim, Also check out Balston Filters (part of the Parker Corporation). The link is : http://www.parker.com/parkersql/default.asp?type=2&id=344
Gary M. Easton Scanners Corporation 410-857-7633
----- Original Message ----- } From: "Vitaly Feingold" {vitalylazar-at-worldnet.att.net} To: {microscopy-at-sparc5.microscopy.com} ; "Jim Romanow" {bsgphy3-at-uconnvm.uconn.edu} Sent: Thursday, April 10, 2003 9:14 PM
Hi, Does anyone know where I could get a pure polystyrene latex for to create a performance specimen as described in "Quality in Electron Microscopy" by Tony Bruton, Steve Chapman, and Paul Harding.
Hi Jim, I frankly don't know what you can do without spending a real bundle. I agree with Vitaly that Edwards BOC is the best bet but you probably have a mixture of pumps. The caveat with respect to the new Edwards oil mist filters is that they fill up with oil, at least on the new 5's and 8's, and if you don't want to be in constant maintenance, you have to purchase the Edwards recirculator packs. Finally, even their refills aren't inexpensive.
For Welch pumps, try: http://www.welchvacuum.com/
For Edwards pumps, try: http://www.bocedwards.com/vacuum/rotary_vane_pump/overview.html
Kurt J. Lesker Vacuum handles about everything you could want and might be the one to give the best prices.
http://www.lesker.com/
Rietschle Pumps can be found here:
http://www.rietschlepumps.com/home.htm
First thing I would recommend is to get a list of Model and serial numbers then you know how much leverage you will have.
Hope this helps, and I do not envy you this task. If it isn't done right, the effort will become a true waste of money as the filter media become saturated and non-functional. Welsh has drains available for their oil mist filters as well. Even when you have the mist depressors, a really good installation will take the air output from the filter to an easily refillable air filter to get the rest of the oil out.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Jim Romanow [mailto:bsgphy3-at-uconnvm.uconn.edu] Sent: Thursday, April 10, 2003 6:43 PM To: microscopy-at-sparc5.microscopy.com
Hello, I must purchase indoor oil mist exhaust filters for six rotary pumps that are attached to EMs, sputter and carbon coaters and a freeze dryer. Can someone please suggest the most practical, easy to maintain economical units from their experience? Thank you. Regards, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
We are trying to measure the proliferation of EPC cells after they have been injected into a host. Prior to injection, we label the cells with CMTMR, a live cell tracker, for identification and subsequently want to do BrdU labelling as a cell proliferation assay.
I fear that the harsh conditions necessary to denature the dsDNA in the BrdU staining protocol will quench the CMTMR signal. First we will test the system in cell culture but even if that works, such conditions may not be as severe as the ones which include paraffin embedding and possible antigen retrieval (cryo sections are probably better) of tissue.
Does anyone have experience with these protocols or are there any suggestions for a more compatible pair of cell marker/proliferation assay products?
Thank you - all suggestions will be highly appreciated.
Judy
Judy Trogadis Bio-Imaging Coordinator St. Michael's Hospital, 8Queen 30 Bond St. Toronto, ON M5B 1W8 Canada ph: 416-864-6060 x6337 pager: 416-685-9219 fax: 416-864-6043 trogadisj-at-smh.toronto.on.ca
The Solid Latex test specimen requires a large quantity (high density) of latex, much more than you can purchase from most suppliers for other applications.
Chuck Garber of Spi has been working on the preparation of a latex test specimen so you could try Spi, or your local university chemistry department. Many of these departments make latex in vast quantities as part of one of their student experiments.
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: "SBC ATC SEM" {atcsem-at-sbcglobal.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Friday, April 11, 2003 3:54 PM
Torino 14 April 2003 (Italy) Hi, I'm an amateur naturalist and I like to improve my optical microscope. So I'm going to change the traditional illumination with tungsten lamp by a LED light source with a 12v and 25 w alimentation (max. 20 mA current at the diode) I noticed that the light is quit strong. I wonder if it wuould be dangerous for my eies, although I use the cyan filter during the observation. Also I can lower the light intensity by a variable resistance. Thank you, With my Best Regards,
I am a PhD student in the Netherlands. I work with the fluorescent tracer Fluoro-Gold. I am trying to do plastic embedding of peripheral nerve and to observe the tracing. However the polymerisation step is performed at 60 deg. C, which is quite detrimental to my signal. Can somebody advise me what kind of plastic should I use. I do not want to heat up my specimens more than 40-45 deg. C.
regards
} Dr. Dimiter Prodanov } Neuroregulation Group, } Department of Neurosurgery, } Leiden University Medical Center, } P.O. Box 9604, 2300 RC Leiden, } The Netherlands } } Tel : +31 71 527 -6760, -6749 } Fax : +31 71 527 6782 }
Greetings, Plastic embedding with UV polymerization allows embedding at 4 C (or even colder). We use a mixture of butyl and methyl methacrylate that is easy and gives good preservation. Its main advantage is that it is extractable with acetone after sectioning so antibodies get great access to their antigens, boosting the signal. The disadvantage is that the resin is a bit soft (no crosslinker) so it is not perfect for TEM use. If you would like a protocol for using butylmethyl methacrylate, contact me off line. Tobias
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Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
Another question: is anyone experiencing what we call "concrete water" in the diamond knife boat??? The symptoms of "concrete water" are that the initial section cuts ok, but subsequent sections pile up becuase they will not move out across the water surface. Even when you manage to separate one section, it is almost impossible to move it about on the surface of the water with a one-hair brush...it just will not move...the water is like "concrete" (Note: this is NOT like the problem of an unclean knife edge when sections pile up for that reason. It is something entirely different.)
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Hello all,
I am a PhD student in the Netherlands. I work with the fluorescent tracer Fluoro-Gold. I am trying to do plastic embedding of peripheral nerve and to observe the tracing. However the polymerisation step is performed at 60 deg. C, which is quite detrimental to my signal. Can somebody advise me what kind of plastic should I use. I do not want to heat up my specimens more than 40-45 deg. C.
regards
} Dr. Dimiter Prodanov } Neuroregulation Group, } Department of Neurosurgery, } Leiden University Medical Center, } P.O. Box 9604, 2300 RC Leiden, } The Netherlands } } Tel : +31 71 527 -6760, -6749 } Fax : +31 71 527 6782 }
} Another question: is anyone experiencing what we call "concrete water" } in the diamond knife boat??? The symptoms of "concrete water" are that } the initial section cuts ok, but subsequent sections pile up becuase } they will not move out across the water surface. Even when you manage } to separate one section, it is almost impossible to move it about on the } surface of the water with a one-hair brush...it just will not } move...the water is like "concrete" (Note: this is NOT like the problem } of an unclean knife edge when sections pile up for that reason. It is } something entirely different.) } } Any ideas???
Yes, I know exactly what you mean: the water behaves like molasses so that the sections are nearly impossible to move. In our experience, we found the cause was a thin film of either protein or oil on the water surface since cleaning the water surface with a lintless lens paper (Rosmarin brand, for example) got rid of the phenomenon (at least temporarily).
You need to eliminate the source of the contamination. This could be traced to either: a dirty knife trough (clean with a gentle detergent), a dirty eyelash probe (caused by cleaning the eyelash by pinching between fingers), dirty glassware that contains the distilled water, contaminated micropore filtration apparatus (change micropore filter and rinse entire apparatus, especially the new micropore filter, with sterile, clean distilled water). Finally, water purification systems (like reverse osmosis or deionization) that become overloaded (or have been recently "recharged") are more likely to cause this than traditionally prepared distilled water.
So, clean everything mentioned. If the problem persists, borrow or purchase some distilled water from a different source and check it out.
############################################################## 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 Fax: 618-453-2665 Email: bozzola-at-siu.edu Web: http://www.siu.edu/~image/ ##############################################################
Does anybody have experience with mito-tracker dye which can distinguish between inactive (dead) mitochondria and active mitochondria. It is probably mostly used with animal or human tissue; however, we would like to use it with corn anthers. The dye seems to have difficulties penetrating into tissue deep enough. If anybody can give me some information or tip, I would be delighted. Thank you in advance. Marianne B. Smith, Schnable-Lab, Agronomy, Iowa State University, Ames, IA 50011
} From: "Massimo" {max_gra-at-libero.it} : : Torino 14 April 2003 : (Italy) : Hi, : I'm an amateur naturalist and I like to improve my optical microscope. : So I'm going to change the traditional illumination with tungsten lamp by a : LED light source with a 12v and 25 w alimentation (max. 20 mA current at the : diode) : I noticed that the light is quit strong. I wonder if it wuould be dangerous : for my eies, although I use the cyan filter during the observation. Also I : can lower the light intensity by a variable resistance.
There are two ways to lower the intensity of a LED. The simplest is to put a variable resistor in series with on leg of the LED and the more complicated is to very the time it turned on and off at frequency of 500 Hz or greater.
Fortunately a LED unlike a conventional bulb does not change color when you limit the current going to it.
Get a data sheet on the LED if you can and see what spectra it puts out. The cyan filter may be redundant. White LEDs are not continuous spectrum devices.
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.
I have received tons of mails from you, which made my mail flow fall into a congested state. Please please delete my e-address fro your list. I do not want to receive e-mail from you. Thank you.
in our lab we have attached to a TEM a Noran Ge-EDX detector with a possibly broken or damaged diamond window. It seems that either the window is broken or that the window is not anymore attached correctly to the counterpart.
Is there anything that we could repair by ourself (either "glueing" or exchang the window)? Does anyone know of any kind of repair kit for such broken windows?
Any advice would be greatly appreciated.
Johannes Bernardi Vienna University of Technology / E052
Back in the days when 4489 was new I too had problems with patchy negatives.
Fortunately Ilford and Kodak were clients so I went along and independently asked them what I was doing wrong. In short almost everything!
They both came up with this procedure
1. Develop sheet film using the dunk and tilt method. 2. Place the film rack in the developer which must be within 2 degs of the ideal 20 deg C if not warm or cool as appropriate. Do not use the time variation to compensate if the temperature is outside this range. 3. After 15 secs lift the film out of the developer and tilt over at 90 degrees for 3 seconds then return to the developer. 4. Repeat the procedure for the desired time, tilting in opposite directions each time. 5. At the development time place the rack of film in and out of water at the same temperature as the developer for 30 seconds. 6. Place the rack of film in the fixer and repeat the dunk and tilt method for the first minute and a half. The said patching could even come from the fixation process; that was new to me.
Problems!
1. The film fogs! Why, because no one replaces safe lights (do they?) and after five years they will have faded and are no longer efficient.
Results!
So good were the negatives that I was able to use a point source enlarger and they are famous for picking out every single defect in a negative! I was running the Hitachi European demo lab at the time, with such cracking pictures we wiped up the London biology market!
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: "bonnie sheppard" {bls4u-at-cstone.net} To: {Microscopy-at-sparc5.microscopy.com} Sent: Monday, April 14, 2003 8:35 PM
Dear Dimiter, } The alternative to heat polymerisation of epoxy resins (e.g. Epon, Spurr) } is to use UV light. Resins used are "London resins" or "Lowicryls", e.g. } "HM20". } Typically this is done after cryofixation and a process called "Freeze } Substition" (FS). During FS the specimen is brought from approx. minus90 } degrees C } to approx. minus30 degrees C ("freeze") while the water in the specimen is } exchanged against solvent ("substitution"). } Once the substitution is complete, infiltration in Lowicryl and } polymerisation under UVlight also happen at approx. -30°C. } The hardened blocks can be sectioned with an ultramicrotome semithin for LM } and ultrathin for TEM. Both types of sections can be immunolabelled. } } There will be a course on "Cryosectioning and Immunolabelling" in Utrecht } this July that will also include a part on cryofixation and FS. } If you need more info, please have a look on the Leica website } "www.em-preparation.com" under "events" or get in contact with Jan Slot and } George Posthuma } in Utrecht: Dr. George Posthuma, Department of Cell Biology } University Medical Center Utrecht, AZU room G.02.525 } Heidelberglaan 100, 3584 CX Utrecht, The Netherlands } Tel. : + 31 30 2506548 } Fax. : + 31 30 2541797 } Email: g.posthuma-at-lab.azu.nl } } } best regards from Vienna, } Joachim } } Dr. Joachim Prutsch } Product Manager EM Specimen Preparation } } Leica Microsystems GmbH } Hernalser Hauptstr. 219 email: } Joachim.Prutsch-at-leica-microsystems.com } A 1170 Vienna Tel.: +43 1 4 88 99 - 235 } AUSTRIA Fax: +43 1 4 88 99 - 350 } } } ------------------------------------------------------------------------ } 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 all, } } I am a PhD student in the Netherlands. I work with the fluorescent tracer } Fluoro-Gold. I am trying to do plastic embedding of peripheral nerve and to } observe the tracing. } However the polymerisation step is performed at 60 deg. C, which is quite } detrimental to my signal. Can somebody advise me what kind of plastic } should } I use. I do not want to heat up my specimens more than 40-45 deg. C. } } regards } } } Dr. Dimiter Prodanov } } Neuroregulation Group, } } Department of Neurosurgery, } } Leiden University Medical Center, } } P.O. Box 9604, 2300 RC Leiden, } } The Netherlands } } } } Tel : +31 71 527 -6760, -6749 } } Fax : +31 71 527 6782
Bonnie, I have not yet actually used the new 4489 film, but spoke with someone at Kodak before purchasing it. Seeing all the negative reports on it, I wanted to "get the scoop" before trying it out. They told me that the best things to try are additional agitation in both the D-19 and Fixer. If the negatives still appear uneven or muddy try using a 1:1 dilution of D-19 instead of the usual 1:2. Good luck to you and anyone else having to use the new film! I hope I won't have too much trouble. Have a good one!
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
-----Original Message----- } From: bonnie sheppard [mailto:bls4u-at-cstone.net] Sent: Monday, April 14, 2003 2:35 PM To: Microscopy-at-sparc5.microscopy.com
Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
Yesterday I posted a message asking if anyone is having trouble with streaking, blotchiness, etc. on kodak 4489 TEM film. I have received no answers, and maybe it is because I did not put my email address in the message.
I have never heard of such a thing. Perhaps one exists, but I doubt it.
The volume behind the window is normally evacuated so special procedures would be needed to replace it. There is also a risk of contamination of the detector crystal if the window is broken. We recently had to send a detector in for repair of a small leak in the window and it was deemed necessary to replace the detector crystal as well. On another occasion years before, just the window was replaced.
In short, I doubt that this project is one that can be done in one's own lab.
I presume most EDS vendors would offer detector repair services. There are also multiple third party vendors available. We have gone both routes. One unit was repaired by Kevex and the other by Gresham. Both were satisfactory.
Warren
At 12:47 PM 4/15/2003 +0200, you wrote:
} Dear all, } } in our lab we have attached to a TEM a Noran Ge-EDX detector with a } possibly broken or damaged diamond window. } It seems that either the window is broken or that the window is not } anymore attached correctly to the counterpart. } } Is there anything that we could repair by ourself (either "glueing" or } exchang the window)? } Does anyone know of any kind of repair kit for such broken windows? } } Any advice would be greatly appreciated. } } Johannes Bernardi } Vienna University of Technology / E052
A vacancy for a technical assistant in the Structural Virology Group of Purdue University is anticipated. This position is ideal for an individual interested in cryo-electron microscopy studies of viruses. The position will involve sample preparation, initial sample assessment, sample freezing, data collection, image analysis and interpretation. A BS with a major in biochemistry or a related area of biology and experience with transmission electron microscopy is considered a minimal requirement. Additional backgrounds in physics and computing along with a willingness to learn and the ability to balance multiple projects would be highly desirable. Employment will entail comprehensive training during the first year and extensive daily interactions with a team of graduate students, post-doctoral scholars and faculty. Our facilities include state of the art 200 and 300kv TEMs. Applications or requests for further information should be sent to Michael Rossmann or Richard Kuhn, Department of Biological Sciences, Purdue University, West Lafayette, Indiana 47907, USA. Tel: 765-494-4911; FAX 765-496-1189; e-mail: mgr-at-indiana.bio.purdue.edu or rjkuhn-at-bragg.bio.purdue.edu.
Paul Chipman Research Electron Microscopist Purdue University Structural Virology
Moxtek definitely sells EDS windows and may sell such a kit. I'm not sure, ask them. www.moxtek.com
Be aware that even if you are successful in gluing in a new window you'll have to re-evacuate (pump out) the cryostat again. That will require a diffusion or turbo pumping station and special adapter to attach it to the detector. A leak detector is also useful.
If you've never undertaken this before it may be better for you in the overall to have an expert do this for you.
Owen
At 12:47 PM 4/15/2003 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Sounds like: 1. some gunk on the block is building up on the knife edge causing the sections to stick. or 2. some contamination in the boat water that is floating on the surface so the sections can't leave the knife edge. 3. Both 1 and 2.
Try cleaning the face and sides of the block, and the knife edge and the boat. Then get some fresh distilled water and a syringe filter to fill the boat.
Geoff
bonnie sheppard wrote:
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-- -- ********************************************** Geoff McAuliffe, Ph.D. Neuroscience and Cell Biology Robert Wood Johnson Medical School 675 Hoes Lane, Piscataway, NJ 08854 voice: (732)-235-4583; fax: -4029 mcauliff-at-umdnj.edu **********************************************
Hi Bonnie, This problem was dealt with recently at length. You should search the archives.
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: bonnie sheppard [mailto:bls4u-at-cstone.net] Sent: Monday, April 14, 2003 3:35 PM To: Microscopy-at-sparc5.microscopy.com
Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
We were also taken by surprise by a box of the "New Formulation" 4489 that we didn't even know we had, until we started using it. This seems to be the film from hell. We solved the density problem by increasing our nitrogen burst agitation to 2 seconds every 10 seconds, but we still are getting strange agitation marks.
I started out years ago using the method described by Steve Chapman, which is the Kodak recommendation, and only started using nitrogen burst when I moved to my present job in 1999. Absolutely no problems until the new film showed up, then we had very thin negatives with horrible streaks. As I said, the density problem has been solved, but we're still fighting the streaks.
What we are doing is a series of exposure trials using "blank" exposed negatives, i.e., exposing the films in the scope with no specimen in place to give an even field of detail-less illumination. This is guaranteed to show any agitation irregularities. We will then vary agitation procedures and times until we get acceptable results. Preliminary results are showing that the plastic film holders with narrow sides, rather then the ones that reach nearly to the bottom of the films, cause fewer agitation problems. Other first results are showing that a combination of "lift and tilt" with nitrogen burst evens things out. Finally, we were getting a "thumb-print" on the bottom of the negatives that turned out to be caused by an agitation swirl around the plastic support rod that holds the films in place. We solved that one by lifting the films up slighty so they don't touch the rod, but that only works on holders that are narrow enough to bend the films when they're inserted.
When we complete our tests, I hope to post images and procedures on our website and post a link to the listserver. In the meantime, I'm following this thread with great interest.
My question is: why would Kodak substitute this for a perfectly good film without any warning? The package insert from a box of the old film was identical to that in the new film, but it seems as if processing procedures need to be radically altered. This has cost us some unhappy clients and no end of headaches, not to mention one embarassing incident where an employee was blamed for something that wasn't his fault. Unless I missed some major press release, this seems like a highly irresponsible action. Does anyone have any insight into this?
Agitatedly, Randy
Randy Tindall EM Core Facility University of Missouri, Columbia
-----Original Message----- } From: Clarkson Donna R Contr USAMRD [mailto:donna.clarkson-at-brooks.af.mil] Sent: Tue 4/15/2003 8:09 AM To: bonnie sheppard; Microscopy-at-sparc5.microscopy.com Cc:
Bonnie, I have not yet actually used the new 4489 film, but spoke with someone at Kodak before purchasing it. Seeing all the negative reports on it, I wanted to "get the scoop" before trying it out. They told me that the best things to try are additional agitation in both the D-19 and Fixer. If the negatives still appear uneven or muddy try using a 1:1 dilution of D-19 instead of the usual 1:2. Good luck to you and anyone else having to use the new film! I hope I won't have too much trouble. Have a good one!
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
-----Original Message----- } From: bonnie sheppard [mailto:bls4u-at-cstone.net] Sent: Monday, April 14, 2003 2:35 PM To: Microscopy-at-sparc5.microscopy.com
Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
I have to make a recommendation whether to buy one or the other cryostat. Do any of you have experience with one or the other or both? Microm is relatively new, gets a lot of recommendations, while Leica, being a long time around seems to receive complaints about service and parts, but we know that Leica is a good microtome. So I would very much like some feedback. Please!! Thank you all in advance.
We are using the new formulation 4489 film, and use nitrogen bursts during developing and fixing (1 second every 10 seconds). We haven't had any problem with fogging, etc. It might be a worthwhile investment to consider...
Marc
On Tuesday, April 15, 2003, at 05:55 AM, Steve Chapman 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 } } Back in the days when 4489 was new I too had problems with patchy } negatives. } } Fortunately Ilford and Kodak were clients so I went along and } independently } asked them what I was doing wrong. In short almost everything! } } They both came up with this procedure } } 1. Develop sheet film using the dunk and tilt method. } 2. Place the film rack in the developer which must be within 2 degs } of } the ideal 20 deg C if not warm or cool as appropriate. Do not use the } time } variation to compensate if the temperature is outside this range. } 3. After 15 secs lift the film out of the developer and tilt over } at 90 } degrees for 3 seconds then return to the developer. } 4. Repeat the procedure for the desired time, tilting in opposite } directions each time. } 5. At the development time place the rack of film in and out of } water at } the same temperature as the developer for 30 seconds. } 6. Place the rack of film in the fixer and repeat the dunk and tilt } method for the first minute and a half. The said patching could even } come } from the fixation process; that was new to me. } } Problems! } } 1. The film fogs! Why, because no one replaces safe lights (do } they?) } and after five years they will have faded and are no longer efficient. } } Results! } } So good were the negatives that I was able to use a point source } enlarger } and they are famous for picking out every single defect in a negative! } I was running the Hitachi European demo lab at the time, with such } cracking } pictures we wiped up the London biology market! } } 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: "bonnie sheppard" {bls4u-at-cstone.net} } To: {Microscopy-at-sparc5.microscopy.com} } Sent: Monday, April 14, 2003 8:35 PM } Subject: Kodak 4489 film } } } } } } ---------------------------------------------------------------------- } } -- } } 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 } } ---------------------------------------------------------------------- } } -. } } } } } } Is anyone else out there having trouble with "shadows and unevenness" } } on } } 4489 film? We have gone to extensive lengths to be sure our } } developing } } is consistent, and still we are having annoying problems with uneven } } (blotchy) negatives. This is not a problem when there is ample tissue } } detail in the negative...but it is horrible and a real printiong } } problem } } for folks looking at DNA spreads, where there is little tissue to } } negative. } } } } Help! } } } } } } } } } } }
-- Marc Pypaert Department of Cell Biology Center for Cell and Molecular Imaging Ludwig Institute for Cancer Research Yale University School of Medicine 333 Cedar Street, PO Box 208002 New Haven, CT 06520-8002 TEL 203-785 3681 FAX 203-785 7446
We are not looking forward to trying the new formulation. Perhaps we will go digital. Can anyone tell me the rough cost of a nitrogen burst system?
Dave
On Tue, 15 Apr 2003 18:53:50 -0400 Marc Pypaert {marc.pypaert-at-yale.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 } -----------------------------------------------------------------------. } } } We are using the new formulation 4489 film, and use nitrogen bursts } during developing and fixing (1 second every 10 seconds). } We haven't had any problem with fogging, etc. } It might be a worthwhile investment to consider... } } Marc } } On Tuesday, April 15, 2003, at 05:55 AM, Steve Chapman 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 } } } } Back in the days when 4489 was new I too had problems with patchy } } negatives. } } } } Fortunately Ilford and Kodak were clients so I went along and } } independently } } asked them what I was doing wrong. In short almost everything! } } } } They both came up with this procedure } } } } 1. Develop sheet film using the dunk and tilt method. } } 2. Place the film rack in the developer which must be within 2 degs } } of } } the ideal 20 deg C if not warm or cool as appropriate. Do not use the } } time } } variation to compensate if the temperature is outside this range. } } 3. After 15 secs lift the film out of the developer and tilt over } } at 90 } } degrees for 3 seconds then return to the developer. } } 4. Repeat the procedure for the desired time, tilting in opposite } } directions each time. } } 5. At the development time place the rack of film in and out of } } water at } } the same temperature as the developer for 30 seconds. } } 6. Place the rack of film in the fixer and repeat the dunk and tilt } } method for the first minute and a half. The said patching could even } } come } } from the fixation process; that was new to me. } } } } Problems! } } } } 1. The film fogs! Why, because no one replaces safe lights (do } } they?) } } and after five years they will have faded and are no longer efficient. } } } } Results! } } } } So good were the negatives that I was able to use a point source } } enlarger } } and they are famous for picking out every single defect in a negative! } } I was running the Hitachi European demo lab at the time, with such } } cracking } } pictures we wiped up the London biology market! } } } } 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: "bonnie sheppard" {bls4u-at-cstone.net} } } To: {Microscopy-at-sparc5.microscopy.com} } } Sent: Monday, April 14, 2003 8:35 PM } } Subject: Kodak 4489 film } } } } } } } } } } ---------------------------------------------------------------------- } } } -- } } } 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 } } } ---------------------------------------------------------------------- } } } -. } } } } } } } } } Is anyone else out there having trouble with "shadows and unevenness" } } } on } } } 4489 film? We have gone to extensive lengths to be sure our } } } developing } } } is consistent, and still we are having annoying problems with uneven } } } (blotchy) negatives. This is not a problem when there is ample tissue } } } detail in the negative...but it is horrible and a real printiong } } } problem } } } for folks looking at DNA spreads, where there is little tissue to } } } negative. } } } } } } Help! } } } } } } } } } } } } } } } } } } } } } -- } Marc Pypaert } Department of Cell Biology } Center for Cell and Molecular Imaging } Ludwig Institute for Cancer Research } Yale University School of Medicine } 333 Cedar Street, PO Box 208002 } New Haven, CT 06520-8002 } TEL 203-785 3681 } FAX 203-785 7446 } }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
Dear SIR, I am Mr.Bliss Marayi and my sister is miss Maureen Marayi we are the children of late Chief John Marayi from Sierra Leone.I am writing you in absolute confidence primarily to seek your assistance to transfer our cash of 18 Million Dollars($18,000.000.00) now in the custody of a private Security trust firm in Europe, the money is in two trunk boxes deposited and declared as family valuables by my late father as a matter of fact the company does not know the content as money, although my father made them to understand that the boxes belongs to his foreign partner. Source of the money: My late father Chief John Marayi , a native of Mende District in the Northerh province of Sierra Leone, was the General Manager of Sierra Leone Mining co-operation (S.L.M.C.) Freetown . According to my father,this money was the income accrued from Mining Co-operation's over draft and minor sales before the peak of the civil war between the rebels forces of Major Paul Koroma and the combined forces of ECOMOG peace keeping operation that almost destroyed my country, following the forceful removal from power of the civilian elected President of Ahmed Tejan Kabbah by the rebels. My father had already made arrangement for his family thats talking about my mother, my little sister and myself to be evacuated to Abidjan Cote d' Ivoire with the CERTIFICATE OF DEPOSIT he made with the security firm in Europe through the aid of U.N evacuation team. During the war in my country, and following the indiscriminate looting of Public and Government properties by the rebel forces, the Sierra Leone mining coop Was one of the target looted and it was destroyed. My father including other top Government functionaries were attacked and killed by the rebels in November 2000 because of his relationship with the civilian Government of Ahmed Tejan Kabbah. As a result of my father's death , and with the news of my uncle's involvement in the air crash in January it dashed our hope of survival.The untimely deaths caused my mother's heart failure and other related complications of which she later died in the hospital after we must have spent a lot of money on her early this year . Now my 18 years old sister and myself are alone in this strange country suffering without any care or help. Without any relation, we are now like refugees and orphans. Our only hope now is in you and the boxes deposited in the Security Firm.To this effect, I humbly solicit your assistance in the followings ways. 1. to assist me claim this boxes from the security Firm as the beneficiary. 2. to transfer this money (USD$18M) in your name to your country. 3. to make a good arrangement for a joint business investment on our behalf in your country and you, our Adviser/ Manager. For your assistance, I have agreed with my younger sister that 20% of the total amount will be for your effort and another 10 % to cover all expenses that may be incured during the business transaction, Lastly, I urge you to keep this transaction strictly confidential as no one knows our where about. Please as you show your willingness, Forward to us your full name,address, Tel and Fax numbers to me via my private email address as indicated bellow, this is for security reasons as i will only be accessing my private email earnestly awaiting your response. Thanks. May God bless you as you assist us.
Dear SIR, I am Mr.Bliss Marayi and my sister is miss Maureen Marayi we are the children of late Chief John Marayi from Sierra Leone.I am writing you in absolute confidence primarily to seek your assistance to transfer our cash of 18 Million Dollars($18,000.000.00) now in the custody of a private Security trust firm in Europe, the money is in two trunk boxes deposited and declared as family valuables by my late father as a matter of fact the company does not know the content as money, although my father made them to understand that the boxes belongs to his foreign partner. Source of the money: My late father Chief John Marayi , a native of Mende District in the Northerh province of Sierra Leone, was the General Manager of Sierra Leone Mining co-operation (S.L.M.C.) Freetown . According to my father,this money was the income accrued from Mining Co-operation's over draft and minor sales before the peak of the civil war between the rebels forces of Major Paul Koroma and the combined forces of ECOMOG peace keeping operation that almost destroyed my country, following the forceful removal from power of the civilian elected President of Ahmed Tejan Kabbah by the rebels. My father had already made arrangement for his family thats talking about my mother, my little sister and myself to be evacuated to Abidjan Cote d' Ivoire with the CERTIFICATE OF DEPOSIT he made with the security firm in Europe through the aid of U.N evacuation team. During the war in my country, and following the indiscriminate looting of Public and Government properties by the rebel forces, the Sierra Leone mining coop Was one of the target looted and it was destroyed. My father including other top Government functionaries were attacked and killed by the rebels in November 2000 because of his relationship with the civilian Government of Ahmed Tejan Kabbah. As a result of my father's death , and with the news of my uncle's involvement in the air crash in January it dashed our hope of survival.The untimely deaths caused my mother's heart failure and other related complications of which she later died in the hospital after we must have spent a lot of money on her early this year . Now my 18 years old sister and myself are alone in this strange country suffering without any care or help. Without any relation, we are now like refugees and orphans. Our only hope now is in you and the boxes deposited in the Security Firm.To this effect, I humbly solicit your assistance in the followings ways. 1. to assist me claim this boxes from the security Firm as the beneficiary. 2. to transfer this money (USD$18M) in your name to your country. 3. to make a good arrangement for a joint business investment on our behalf in your country and you, our Adviser/ Manager. For your assistance, I have agreed with my younger sister that 20% of the total amount will be for your effort and another 10 % to cover all expenses that may be incured during the business transaction, Lastly, I urge you to keep this transaction strictly confidential as no one knows our where about. Please as you show your willingness, Forward to us your full name,address, Tel and Fax numbers to me via my private email address as indicated bellow, this is for security reasons as i will only be accessing my private email earnestly awaiting your response. Thanks. May God bless you as you assist us.
I am having an odd problem staining cell monolayers with a directly labled FITC antibody to an intracellular antigen. If I try to stain the cells in either a labtek slide chamber (glass) or in a 96 well plate I get no staining. If I remove the cells with trypsin and make a cytospin the cells stain perfectly.
I really need to stain the intact monolayers for routine work, but nothing I have tried seems to work.
Some of the permutations I have tried include:
Fixing with PFA then permeabilizing with either Triton or MeOH then stain
Remove the media and let the monolayer AIR DRY, then permeabilizing with either Triton or MeOH then stain
Fix/permeabilize the wet monolayer with MeOH then stain
Suggestions? --
Michael J. Herron, U of MN, Entomology herro001-at-umn.edu 612-624-3688 Office, 624-3212 Lab, 625-5299 FAX
Hi Marianne, I have been using a Leica cryostat, motorized. It is really good and I am happy with it. For the parts, I can't say anything because it is new and therefore didn't need any service. In my area (Montreal, Quebec) the tech rep is really good but very busy, but so far I he always gave a good service (for other equipments). I guess it depends on who is in your area. Hope this helps! Emmanuelle
Randy, I'm sure it's got something to do with Kodak saving a fraction of a penny per film box. Remember Coca-Cola's new formulation years ago? Seems like Kodak should've taken a lesson. There's a Murphy's Law corollary which states, " If it ain't broke, don't fix it!"
WWW ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Winston W. Wiggins, Supervisor Cannon Electron Microscopy Lab Carolinas Medical Center P.O. Box 32861; Charlotte, NC 28232-2861 Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-0589 E-mail: {mailto:WWiggins-at-Carolinas.org} Winston.Wiggins-at-CarolinasHealthCare.org ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
-----Original Message----- } From: Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] Sent: Tuesday, April 15, 2003 1:33 PM To: Clarkson Donna R Contr USAMRD Cc: microscopy-at-sparc5.microscopy.com
Hi,
We were also taken by surprise by a box of the "New Formulation" 4489 that we didn't even know we had, until we started using it. This seems to be the film from hell. We solved the density problem by increasing our nitrogen burst agitation to 2 seconds every 10 seconds, but we still are getting strange agitation marks.
I started out years ago using the method described by Steve Chapman, which is the Kodak recommendation, and only started using nitrogen burst when I moved to my present job in 1999. Absolutely no problems until the new film showed up, then we had very thin negatives with horrible streaks. As I said, the density problem has been solved, but we're still fighting the streaks.
What we are doing is a series of exposure trials using "blank" exposed negatives, i.e., exposing the films in the scope with no specimen in place to give an even field of detail-less illumination. This is guaranteed to show any agitation irregularities. We will then vary agitation procedures and times until we get acceptable results. Preliminary results are showing that the plastic film holders with narrow sides, rather then the ones that reach nearly to the bottom of the films, cause fewer agitation problems. Other first results are showing that a combination of "lift and tilt" with nitrogen burst evens things out. Finally, we were getting a "thumb-print" on the bottom of the negatives that turned out to be caused by an agitation swirl around the plastic support rod that holds the films in place. We solved that one by lifting the films up slighty so they don't touch the rod, but that only works on holders that are narrow enough to bend the films when they're inserted.
When we complete our tests, I hope to post images and procedures on our website and post a link to the listserver. In the meantime, I'm following this thread with great interest.
My question is: why would Kodak substitute this for a perfectly good film without any warning? The package insert from a box of the old film was identical to that in the new film, but it seems as if processing procedures need to be radically altered. This has cost us some unhappy clients and no end of headaches, not to mention one embarassing incident where an employee was blamed for something that wasn't his fault. Unless I missed some major press release, this seems like a highly irresponsible action. Does anyone have any insight into this?
Agitatedly, Randy
Randy Tindall EM Core Facility University of Missouri, Columbia
-----Original Message----- } From: Clarkson Donna R Contr USAMRD } [ {mailto:donna.clarkson-at-brooks.af.mil} mailto:donna.clarkson-at-brooks.af.mil] Sent: Tue 4/15/2003 8:09 AM To: bonnie sheppard; Microscopy-at-sparc5.microscopy.com Cc:
Bonnie, I have not yet actually used the new 4489 film, but spoke with someone at Kodak before purchasing it. Seeing all the negative reports on it, I wanted to "get the scoop" before trying it out. They told me that the best things to try are additional agitation in both the D-19 and Fixer. If the negatives still appear uneven or muddy try using a 1:1 dilution of D-19 instead of the usual 1:2. Good luck to you and anyone else having to use the new film! I hope I won't have too much trouble. Have a good one!
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
-----Original Message----- } From: bonnie sheppard [ {mailto:bls4u-at-cstone.net} mailto:bls4u-at-cstone.net] Sent: Monday, April 14, 2003 2:35 PM To: Microscopy-at-sparc5.microscopy.com
Is anyone else out there having trouble with "shadows and unevenness" on 4489 film? We have gone to extensive lengths to be sure our developing is consistent, and still we are having annoying problems with uneven (blotchy) negatives. This is not a problem when there is ample tissue detail in the negative...but it is horrible and a real printiong problem for folks looking at DNA spreads, where there is little tissue to negative.
Help!
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We, at the high voltae microscope had used 4489 for years. A few years ago we noticed streaking, particulary from top to bottom. Attributing it to the way the films are held verical in the developing tank and that for the most part the nitrogen burst bubbles followed thesame paths repeatedly; we stopped the bursting for most of the developing time. Instead we would move the racks up and down and sideways so that the flow of developer was for the most part parallel to the film. This seemed to help but it took a lot more care and time.
As far as digital goes, I would suggest that film, particularly 4489, can give much more information and micrograph quality than almost any digital systems.
What you gain with digital is time efficiency ( no developing), better quantitation (CCDs are very linear with exposure), and perhapse more sensativity. The results are not subject to variation and whims of the development process. But what you loose with digital is the large uniform field of information ( 4489 would exceed 5000x5000 resolution easily), Digital tends to have more grainieness due to the scintillator and fiber optic transfer plates. If you are using 4489 instead of SO163 you are probably trying to get a very fine grained smooth image - going digital is going the other way. The dynamic range of 4489 film is greater than all but the most spectacular digital systems (probably unaffordable). This effects how well you can resolve micrographs with high brightness relief - with bright areas of detail and dark areas on the same frame.
If you have the time film is good.
Dave -- David Barnard Wadsworth Ctr NYS Dept Health Albany NY 12201-0509 barnard-at-wadsworth.org 518 473-5299 voice 518 474-7992 fax
I would like to say "Thank you very much" to all the people who took the time to respond to my request for information about and experience with the various 120kV TEMs.
This is Lifeng Dong from Department of Physics, Portland State University, USA. I am planning to attend M&M 2003, and reserved a double room (two beds) from Hampton Inn Downtown San Antonio. The price including tax is $121.42 per night, which is a little expensive for a graduate student. If you or some of your male colleagues will attend this conference and are looking for a hotel, I would like to share it with you or him.
If you have any question, contact me at lifengd-at-pdx.edu.
Hi, I want to thanks all answered my question about illumination by LED and at the same time I want to assure about my project that I am not going to use a Laser LED but a simple led lamp just like you are get used to see on some device with colors red, green. The difference is that my light is white. In spite of that I think would be better to insert an infrared filter or similar as you have recommended me. The device that I'm making is like that you can find at this address: http://www.microscopy-uk.org.uk/mag/artjul99/bdoumic.html Thank you. With my Best Regards,
Cost depends on what you already have. A gas burst timer is approximately $400.00. A setup with 3-1gal tanks in a water jacket with floating lids, a cover, and plenums in two tanks would be approximately $1100. This does not include the timer or water temperature control. A similar setup with 3-3.5gal tanks would be approximately $1400. These are estimates..
George
George Laing National Graphic Supply 800.223.7130 x3109 scisales-at-ngscorp.com
} } We are not looking forward to trying the new formulation. } Perhaps we will go digital. Can anyone tell me the rough } cost of a nitrogen burst system? } } Dave
I have the option to buy either a 5 or 3 megapixel camera. What I want to know is whether a 5 megapixel camera is better than a 3 megapixel camera, if used exclusively on a microscope, or whether the extra pixels (and cost) are wasted. Assume bit depth is the same.
I'd like someone to check my math and my reasoning.
I understand that microscope resolution is limited by diffraction and a good rule of thumb for displaying images at normal reading distance is a final image mag of no more than 1000x the numerical aperture of the objective. On my microscope this means about 100x for images taken with the 4x lens and 1250x for images with the 100x oil lens. Assuming I use a 1x relay eyepiece to connect the CCD camera, the image at the detector could be magnified 25 times for the 4x objective, and 12.5 times for the 100x objective, before getting into empty mag.
On the camera side, I understand that the more pixels the better the resolution. The 3 megapixel camera has 2080 horizontal pixels and the 5mp has 2580. Let's ignore the vertical pixels for the moment.
One more assumption is that for printing, it seems that the highest practical output resolution is 266-300 dpi. This means that the 3 mp camera produces an image that optimally prints an image 2080/266 = 7.8 inches or 198 mm horizontally, or less, and the 5 mp can print an image about 243 mm horizontally or less. Enlarging above these dimensions is possible but the print image will need interpolation and might be degraded.
Going back to the microscope, the maximum horizontal field of view with the 4x objective captured by the camera is 1.6 mm. It could be printed up to 100 x 1.6 = 160 mm before empty mag sets in. But I have already calculated that the 3 mp can print up to 198 mm and the 5 mp up to 243 mm before the print image degrades.
If these calculations are correct (please tell me if it they are not) then both the 3 mp and 5 mp have more than enough resolution to capture what the microscope makes available at 4x, and the situation is even better for the 100x lens because the lens does most of the heavy lifting, magnification-wise.
One final condition that might matter is that both are RBG cameras with Bayer filters, so that the real resolution is approximately 1/3 of the nominal pixel resolution. I'm not sure how to figure that into the calculations.
So, at approximately a dollar-per-pixel, do I need the extra 2000 pixels of a 5 mp camera?
Gary P. Radice gradice-at-richmond.edu Department of Biology 804-289-8107 (voice) University of Richmond 804-289-8233 (FAX) Richmond VA 23173 http://www.richmond.edu/~gradice
When teaching an EM course, how many additional non-EM courses might others teach per semester? I am presently teaching 2 courses, one of which is on SEM. I also teach microtechnique and a TEM course.
Obviously, the answer depends on EM course formats and campus size.
My courses are intensive, usually involving 5-6 students, sometimes more. An SEM course may take 15-25 hours per week, including all formally scheduled lab and lecture times, weekly tutorials, solutions preparation etc.(there is no assistant). Our campus is small, perhaps 14,000 students.
Since I teach so few students, my Chair is wondering about the fairness of my workload relative to others in my department who teach many more students in straight lecture courses (the larger courses have assistants).
Your answers will be most helpful in clarifying a "normal" academic work load that includes EM instruction at other similar sized universities. Please send your replies to me at the e-mail address below. I will summarize the results and post them on the listserver.
Thank you in advance!
Jerry
--------------------- Jerry D. Kudenov Dept. Biological Sciences University of Alaska Anchorage 3211 Providence Drive Anchorage, Alaska 99508
} My question is: why would Kodak substitute this for a perfectly good film } without any warning? The package insert from a box of the old film was } identical to that in the new film, but it seems as if processing } procedures need to be radically altered. This has cost us some unhappy } clients and no end of headaches, not to mention one embarrassing incident } where an employee was blamed for something that wasn't his fault. Unless I } missed some major press release, this seems like a highly irresponsible } action. Does anyone have any insight into this? } } Agitatedly, } Randy
Normally I don't like litigation but the pain and suffering and the lack of warning seems to point to a class action suit, and at least an official explanation accompanying the new film and properly labeled new film. I'm maybe just surprised to not have seen that suggestion in the previous discussion on 4489 formula change.
I know this has been gone over in length before. We do not have any nitrogen capabilities in our darkroom, and I am dreading the next batch of 4489 film. Maybe Kodak is using this like Coca-cola did when they made "new" coke, and then switched back to "classic" but that change let them go from using expensive beet sugar to corn sugar as the primary sweetener saving millions of dollars. Who knows - but from the great discussion here I at least have a plan for when we get the new formula (not that I will be able to implement it).
Geoff Williams Microscopy Facility Supervisor
CMU Biology Department Microscopy Facility web page. http://www.cst.cmich.edu/centers/microscopy/
Hello Gary Your calculations sounds reasonable. The things you always have to keep in mind going "digital" are that most problems appeared when you need to crop (and possible magnify after all) your image in order to show some small details etc. It happening frequently when image prepared for publication. There you will face the real problem with amount of pixels you have. Sergey
At 11:30 AM 4/16/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
On Wednesday, April 16, 2003, at 03:16 AM, Patton, David wrote:
} We are not looking forward to trying the new formulation. } Perhaps we will go digital. Can anyone tell me the rough } cost of a nitrogen burst system? } Dear David, We just got one from Pella for ~$1150. 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
In respond on my message on the "first wave" of the 4489 new formulation discussion I got respond from Kodak that most customers are very happy with new formulation and ONLY a few were unhappy. I am too lazy to find that message in my archive. I wrote back asking why they did not change the product name if altering development process on "new formulation" - I never had respond back. Sergey
At 02:12 PM 4/16/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (ramg21-at-yahoo.co.in) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, April 16, 2003 at 11:36:56 ---------------------------------------------------------------------------
Email: ramg21-at-yahoo.co.in Name: ram varma
Organization: univ of louisiana lafayette
Education: Graduate College
Location: lafayette,LA, USA
Question: could you tell me where i can find comprehensive information about
1] near field optical microscopy 2] reflectometry 3] ellipsometry
I would like to know all the details viz. equipment specification,opration,theory of operation etc.
Sounds like the paint crew I hired once to paint my house- all other customers were always happy, no further response.
Vitaly Feingold Scientific Instruments and Applications 2773 Heath Lane, Duluth GA 30096 (770)232-7785 ph. (770)232-1791 fax (678)467-0012 mobile
This message is made of 100% recycled electrons.
This address can not receive messages larger than 15 kb without prior notification.
----- Original Message ----- } From: "Sergey Ryazantsev" {sryazant-at-ucla.edu} To: {Microscopy-at-sparc5.microscopy.com} Sent: Wednesday, April 16, 2003 10:05 PM
} From: "Massimo" {max_gra-at-libero.it} : : Torino 16 April 2003 (ITALY) : : Hi, : I want to thanks all answered my question about illumination by LED and at : the same time I want to assure about my project that I am not going to use a : Laser LED but a simple led lamp just like you are get used to see on some : device with colors red, green. The difference is that my light is white. : In spite of that I think would be better to insert an infrared filter or : similar as you have recommended me. : The device that I'm making is like that you can find at this address: : http://www.microscopy-uk.org.uk/mag/artjul99/bdoumic.html
That will work fine. The LED you are using will have a rating in milliamps or 1/1000 amps. The resistor R1 is what limits the current to keep from burning the LED out. The value of R1 is found R1=Voltage divided by the maximum current that the LED can carry.
e.g. If you have 20 ma LED and are using 5 volts R1=5/.02 = 250 ohms. The variable resistor should be a 1,000 ohms. If you are using a ultra bright LED you need to consider the power that the resistor carries that is represented by the voltage times the current and is represented in watts in this case it is 250 *.02 = 0.1 watt so a .25 watt resistor will do. An ultra bright LED will need a half watt or possibly a 1 watt resistor in both R1 and R2.
Any voltage over 3 volts will work. With a 1.5 volt battery you don't need R1 in most cases and with less than 3 volts you may not get full brightness from the LED.
None of the LED materials that emit in the visible range emit infra red see: http://www.theledlight.com/technical3.html
Here is a lot more information on LED construction. http://www.theledlight.com/technical.html
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.
Having read of the problems experienced by many users of Kodak 4489 film, after the formulation or something else in the production process appears to have changed, we faced using our new stock with much fear and trepidation. Therefore, in order to try to see what effect this would have in our lab, and to see what we could do to minimize the problems faced by other users, I decided to fill a film cassette half with "old" 4489 film and half with the "new" film and then instructed our darkroom technician to process the film exactly as he had always done, but to increase manual agitation of the film (we do not have nitrogen burst agitation) to 10 seconds at a time at 10 second intervals throughout development, i.e. 3 x 10 second agitations per minute.
After development the difference between the "old" and the "new" was immediately obvious, but only in that the "new" negatives were significantly denser (darker) than the old. There does not appear to be any steaking or inconsistency in the development that others have reported. Although the "new" negatives are not so dark as to be difficult to print, I have since adjusted the TEM's film sensitivity setting to allow us to obtain negatives that are similar in density to those that we obtained from the previous 4489 film.
Perhaps the fact that we do not experience the same problems as reported by others has something to do with the development conditions that we use? We use Ilford PQ Universal developer diluted 1 + 9 for 5 minutes at 20C.
I hope this does not further confuse the issue!
Regards
Rob
=====================================
Rob Cross Director : EM Unit, Rhodes University tel: (046) 603 8168/9
We also use Ilford PQ Universal with Kodak film, 4489 and SO-163, because it is much more convenient. We only use D-19 very very occaisionally for a forced development protocol.
Sally Stowe
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University, Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 http://www.anu.edu.au/EMU
} } } "R. Cross" {r.cross-at-ru.ac.za} 04/17/03 05:10PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Dear 4489 users
Having read of the problems experienced by many users of Kodak 4489 film, after the formulation or something else in the production process appears to have changed, we faced using our new stock with much fear and trepidation. Therefore, in order to try to see what effect this would have in our lab, and to see what we could do to minimize the problems faced by other users, I decided to fill a film cassette half with "old" 4489 film and half with the "new" film and then instructed our darkroom technician to process the film exactly as he had always done, but to increase manual agitation of the film (we do not have nitrogen burst agitation) to 10 seconds at a time at 10 second intervals throughout development, i.e. 3 x 10 second agitations per minute.
After development the difference between the "old" and the "new" was immediately obvious, but only in that the "new" negatives were significantly denser (darker) than the old. There does not appear to be any steaking or inconsistency in the development that others have reported. Although the "new" negatives are not so dark as to be difficult to print, I have since adjusted the TEM's film sensitivity setting to allow us to obtain negatives that are similar in density to those that we obtained from the previous 4489 film.
Perhaps the fact that we do not experience the same problems as reported by others has something to do with the development conditions that we use? We use Ilford PQ Universal developer diluted 1 + 9 for 5 minutes at 20C.
I hope this does not further confuse the issue!
Regards
Rob
=====================================
Rob Cross Director : EM Unit, Rhodes University tel: (046) 603 8168/9
you are right. From the point of microscopy there are only two reasons to have more pixels:
#1 To print of large images (e.g. for posters or for other representations) without loss of details. #2 For digital processing of your images (e.g. to 'improve' resolution of instrument or to extract details).
Or are there other reasons?
} From a major interest should be the data depth (possible grey steps in each pixel) for a computer processing of brightness and contrast. If you have high dynamics, even wrong lighted images are able to convert into better results or you are able to improve contrast of low differences in an image region, which is uniform in colour and brightness with first views.
Frank Eggert
http://www.microanalyst.net
Gary Radice schrieb:
} ------------------------------------------------------------------------ } 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 the option to buy either a 5 or 3 megapixel camera. What I want } to know is whether a 5 megapixel camera is better than a 3 megapixel } camera, if used exclusively on a microscope, or whether the extra } pixels (and cost) are wasted. Assume bit depth is the same. } } I'd like someone to check my math and my reasoning. } } I understand that microscope resolution is limited by diffraction and a } good rule of thumb for displaying images at normal reading distance is } a final image mag of no more than 1000x the numerical aperture of the } objective. On my microscope this means about 100x for images taken with } the 4x lens and 1250x for images with the 100x oil lens. Assuming I use } a 1x relay eyepiece to connect the CCD camera, the image at the } detector could be magnified 25 times for the 4x objective, and 12.5 } times for the 100x objective, before getting into empty mag. } } On the camera side, I understand that the more pixels the better the } resolution. The 3 megapixel camera has 2080 horizontal pixels and the } 5mp has 2580. Let's ignore the vertical pixels for the moment. } } One more assumption is that for printing, it seems that the highest } practical output resolution is 266-300 dpi. This means that the 3 mp } camera produces an image that optimally prints an image 2080/266 = 7.8 } inches or 198 mm horizontally, or less, and the 5 mp can print an image } about 243 mm horizontally or less. Enlarging above these dimensions is } possible but the print image will need interpolation and might be } degraded. } } Going back to the microscope, the maximum horizontal field of view with } the 4x objective captured by the camera is 1.6 mm. It could be printed } up to 100 x 1.6 = 160 mm before empty mag sets in. But I have already } calculated that the 3 mp can print up to 198 mm and the 5 mp up to 243 } mm before the print image degrades. } } If these calculations are correct (please tell me if it they are not) } then both the 3 mp and 5 mp have more than enough resolution to capture } what the microscope makes available at 4x, and the situation is even } better for the 100x lens because the lens does most of the heavy } lifting, magnification-wise. } } One final condition that might matter is that both are RBG cameras with } Bayer filters, so that the real resolution is approximately 1/3 of the } nominal pixel resolution. I'm not sure how to figure that into the } calculations. } } So, at approximately a dollar-per-pixel, do I need the extra 2000 } pixels of a 5 mp camera? } } Gary P. Radice gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond 804-289-8233 (FAX) } Richmond VA 23173 http://www.richmond.edu/~gradice
At least with Coke there are other soft drinks. As far as I can tell 4489 is the only film available for routine biological tissue work. I have formed the impression from this listserver that S0163 (?) is for more specialised use. Therefore I think we are over a barrel re voting with our feet (mixed metaphors anyone?).
} ------------------------------------------------------------------------ } 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 } -----------------------------------------------------------------------. } } } Randy, } I'm sure it's got something to do with Kodak saving a fraction of a } penny per film box. Remember Coca-Cola's new formulation years ago? } Seems like Kodak should've taken a lesson. There's a Murphy's Law } corollary which states, " If it ain't broke, don't fix it!" } } WWW } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W. Wiggins, Supervisor } Cannon Electron Microscopy Lab } Carolinas Medical Center } P.O. Box 32861; Charlotte, NC 28232-2861 } Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 } Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-0589 } E-mail: {mailto:WWiggins-at-Carolinas.org} Winston.Wiggins-at-CarolinasHealthCare.org } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } -----Original Message----- } } From: Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] } Sent: Tuesday, April 15, 2003 1:33 PM } To: Clarkson Donna R Contr USAMRD } Cc: microscopy-at-sparc5.microscopy.com } Subject: RE: Kodak 4489 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Hi, } } We were also taken by surprise by a box of the "New Formulation" 4489 } that we didn't even know we had, until we started using it. This } seems to be the film from hell. We solved the density problem by } increasing our nitrogen burst agitation to 2 seconds every 10 } seconds, but we still are getting strange agitation marks. } } I started out years ago using the method described by Steve Chapman, } which is the Kodak recommendation, and only started using nitrogen } burst when I moved to my present job in 1999. Absolutely no problems } until the new film showed up, then we had very thin negatives with } horrible streaks. As I said, the density problem has been solved, } but we're still fighting the streaks. } } What we are doing is a series of exposure trials using "blank" } exposed negatives, i.e., exposing the films in the scope with no } specimen in place to give an even field of detail-less illumination. } This is guaranteed to show any agitation irregularities. We will } then vary agitation procedures and times until we get acceptable } results. Preliminary results are showing that the plastic film } holders with narrow sides, rather then the ones that reach nearly to } the bottom of the films, cause fewer agitation problems. Other first } results are showing that a combination of "lift and tilt" with } nitrogen burst evens things out. Finally, we were getting a } "thumb-print" on the bottom of the negatives that turned out to be } caused by an agitation swirl around the plastic support rod that } holds the films in place. We solved that one by lifting the films up } slighty so they don't touch the rod, but that only works on holders } that are narrow enough to bend the films when they're inserted. } } When we complete our tests, I hope to post images and procedures on } our website and post a link to the listserver. In the meantime, I'm } following this thread with great interest. } } My question is: why would Kodak substitute this for a perfectly good } film without any warning? The package insert from a box of the old } film was identical to that in the new film, but it seems as if } processing procedures need to be radically altered. This has cost us } some unhappy clients and no end of headaches, not to mention one } embarassing incident where an employee was blamed for something that } wasn't his fault. Unless I missed some major press release, this } seems like a highly irresponsible action. Does anyone have any } insight into this? } } Agitatedly, } Randy } } Randy Tindall } EM Core Facility } University of Missouri, Columbia } } } } } } -----Original Message----- } } From: Clarkson Donna R Contr USAMRD } } [ {mailto:donna.clarkson-at-brooks.af.mil} mailto:donna.clarkson-at-brooks.af.mil] } Sent: Tue 4/15/2003 8:09 AM } To: bonnie sheppard; Microscopy-at-sparc5.microscopy.com } Cc: } Subject: RE: Kodak 4489 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Bonnie, } I have not yet actually used the new 4489 film, but spoke with someone } at Kodak before purchasing it. Seeing all the negative reports on it, I } wanted to "get the scoop" before trying it out. They told me that the best } things to try are additional agitation in both the D-19 and Fixer. If the } negatives still appear uneven or muddy try using a 1:1 dilution of D-19 } instead of the usual 1:2. } Good luck to you and anyone else having to use the new film! I hope I } won't have too much trouble. } Have a good one! } } 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 } } } -----Original Message----- } } From: bonnie sheppard [ {mailto:bls4u-at-cstone.net} mailto:bls4u-at-cstone.net] } Sent: Monday, April 14, 2003 2:35 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Kodak 4489 film } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver } On-Line Help } {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } -----------------------------------------------------------------------. } } } Is anyone else out there having trouble with "shadows and unevenness" on } 4489 film? We have gone to extensive lengths to be sure our developing } is consistent, and still we are having annoying problems with uneven } (blotchy) negatives. This is not a problem when there is ample tissue } detail in the negative...but it is horrible and a real printiong problem } for folks looking at DNA spreads, where there is little tissue to } negative. } } Help! } } } } } } } } } *********************************************************************** } This electronic message may contain information that is confidential } and/or legally privileged. It is intended only for the use of the } individual(s) and entity named as recipients in the message. If you } are not an intended recipient of this message, please notify the } sender immediately and delete the material from any computer. Do not } deliver, distribute or copy this message, and do not disclose its } contents or take any action in reliance on the information it } contains. Thank you. }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
The first place to start is a Google search on each of the terms. A similar search could be done under PubMed (National Library of Medicine). I have found that the Google searches find good on-line references from educational institutions that range from the basic to in-depth.
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 } -----------------------------------------------------------------------. } } } Below is the result of your feedback form (NJZFM-ultra-55). It was } submitted by (ramg21-at-yahoo.co.in) from } http://www.msa.microscopy.com/Ask-A-Microscopist.html on Wednesday, } April 16, 2003 at 11:36:56 } --------------------------------------------------------------------------- } } Email: ramg21-at-yahoo.co.in } Name: ram varma } } Organization: univ of louisiana lafayette } } Education: Graduate College } } Location: lafayette,LA, USA } } Question: could you tell me where i can find comprehensive information about } } 1] near field optical microscopy } 2] reflectometry } 3] ellipsometry } } I would like to know all the details viz. equipment } specification,opration,theory of operation etc. } } --------------------------------------------------------------------------- }
Capturing the spatial resolution of your microscope is a matter of both inner and outer resolution. The inner resolution is the capability of you camera to resolve the spatial information provided by your microscope. This is determined by the spatial resolution of the microscope, total magnification and the pixel size of the CCD-grid on your camera. The Nyquist sampling theorem and the Whittaker-Shannon Sampling theorem deals with this issue:
The larger the individual pixels of the camera, the more you will need to magnify the image, given the same spatial resolution. This is important for quantification, but can be more relaxed for visualisation or publications.
The outer resolution is determined by the fraction of the field of view the camera can capture on its CCD. The larger the field covered by the camera in one image the less you will need to move the microscope stage to view more of the sample.
So in a sense the digital image is defined by its inner and outer resolution. This is only a short description of this issue and there are of course more and more precise thing to say about it.
===================================================== } } I have the option to buy either a 5 or 3 megapixel camera. What I want to } } know is whether a 5 megapixel camera is better than a 3 megapixel } } camera, if used exclusively on a microscope, or whether the extra pixels } } (and cost) are wasted. Assume bit depth is the same. } } } } I'd like someone to check my math and my reasoning. } } } } I understand that microscope resolution is limited by diffraction and a } } good rule of thumb for displaying images at normal reading distance is a } } final image mag of no more than 1000x the numerical aperture of the } } objective. On my microscope this means about 100x for images taken with } } the 4x lens and 1250x for images with the 100x oil lens. Assuming I use a } } 1x relay eyepiece to connect the CCD camera, the image at the detector } } could be magnified 25 times for the 4x objective, and 12.5 times for the } } 100x objective, before getting into empty mag. } } } } On the camera side, I understand that the more pixels the better the } } resolution. The 3 megapixel camera has 2080 horizontal pixels and the 5mp } } has 2580. Let's ignore the vertical pixels for the moment. } } } } One more assumption is that for printing, it seems that the highest } } practical output resolution is 266-300 dpi. This means that the 3 mp } } camera produces an image that optimally prints an image 2080/266 = 7.8 } } inches or 198 mm horizontally, or less, and the 5 mp can print an image } } about 243 mm horizontally or less. Enlarging above these dimensions is } } possible but the print image will need interpolation and might be degraded. } } } } Going back to the microscope, the maximum horizontal field of view with } } the 4x objective captured by the camera is 1.6 mm. It could be printed up } } to 100 x 1.6 = 160 mm before empty mag sets in. But I have already } } calculated that the 3 mp can print up to 198 mm and the 5 mp up to 243 mm } } before the print image degrades. } } } } If these calculations are correct (please tell me if it they are not) then } } both the 3 mp and 5 mp have more than enough resolution to capture what } } the microscope makes available at 4x, and the situation is even better for } } the 100x lens because the lens does most of the heavy lifting, } } magnification-wise. } } } } One final condition that might matter is that both are RBG cameras with } } Bayer filters, so that the real resolution is approximately 1/3 of the } } nominal pixel resolution. I'm not sure how to figure that into the } } calculations. } } } } So, at approximately a dollar-per-pixel, do I need the extra 2000 pixels } } of a 5 mp camera? } } } } } } Gary P. Radice gradice-at-richmond.edu } } Department of Biology 804-289-8107 (voice) } } University of Richmond 804-289-8233 (FAX) } } Richmond VA 23173 http://www.richmond.edu/~gradice } }
I suggest that everyone unhappy with the new formulation of 4489 should contact Eric Ambrose, and respond to his post of Jan 24th.
Jim
PS: Our users are some of the "few" unhappy.
} } Eric Ambrose } } eric.ambrose-at-kodak.com } } Silver Halide Product Manager } } Scientific Imaging Systems } } Eastman Kodak Company
Jim
Sergey wrote: } } In respond on my message on the "first wave" of the 4489 new formulation } discussion I got respond from Kodak that most customers are very happy with } new formulation and ONLY a few were unhappy. I am too lazy to find that } message in my archive. I wrote back asking why they did not change the } product name if altering development process on "new formulation" - I never } had respond back. Sergey } } }
} } From Microscopy-request-at-sparc5.microscopy.com Fri Jan 24 05:38:36 2003 } } Date: Thu, 23 Jan 2003 18:25:06 -0600 } } To: Microscopy-at-sparc5.microscopy.com } } From: eric.ambrose-at-kodak.com (by way of MicroscopyListserver) } } Subject: TEM Imaging: Kodak 4489 EM Film Reformulation } } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } To Subscribe/Unsubscribe -- Send Email to ListServer-at-MSA.Microscopy.Com } } On-Line Help http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html } } -----------------------------------------------------------------------. } } } } } } } } } } Subject: TEM Imaging: Kodak 4489 EM Film Reformulation } } } } Recently, there have been a number of messages posted on the MSA ListServer } } regarding issues with the reformulated KODAK Electron Microscope Film 4489. } } As the Kodak product manager for the films for electron micrography, I } } would like to offer the following information in response to the recent } } inquiries on Kodak EM films. } } } } Scientific Imaging Systems, a division of Eastman Kodak Company, remains } } committed to supplying the highest quality scientific imaging products } } required to meet the critical needs of today's research scientists. Kodak } } manufactures and sells two different films for use in TEM imaging, KODAK } } Electron Microscope Film 4489 and KODAK Electron Image Film SO-163. } } } } Over the years, for many different reasons, Kodak has made changes to the } } formulations of these films. Some changes have been minor while others have } } been more significant. The decision to make a formulation change is one of } } the most contentious and difficult decisions that we address. Most of } } these changes are forced upon us in response to manufacturing equipment } } requirements, market demand, and/or issues with supply of raw material. } } Whatever the reason, Kodak makes these required changes not only to ensure } } the continued availability of electron micrography products to scientific } } customers, but also to ensure the products perform to the highest quality } } standards. Every change to a formulation is accompanied by extensive } } in-house and customer testing to ensure the product meets its } } specifications. Various techniques performed in the research community can } } result in some instances where modifications to some protocols are required } } to achieve comparable results with a reformulated film compared to its } } predecessor. } } } } Scientific Imaging Systems (SIS) has a dedicated team of technical support } } staff ready to assist anyone with questions or concerns about the } } performance of the Kodak EM films, as well as any other products sold by } } SIS. People should not hesitate to contact the SIS Technical Support Group } } with questions, concerns, and suggestions. We value our customers and } } will make every effort to assist you. Please email us at: } } sis-support-at-kodak.com or call 1-877-SIS-HELP or 203-786-5657. } } } } In regards to some of the comments and issues recently expressed on the MSA } } Microscopy ListServer, I have personally contacted, or have attempted to } } contact, each person listed with a concern. As in many instances, we need } } to see the films themselves to make a determination on the issue and how to } } best assist the customer going forth. Most customers that have used the } } reformulated 4489 film have found that it supplies the same high quality } } results they have always received from Kodak EM films. However, as seen on } } the Microscopy ListServer, we do have a small number of customers that } } appear to have an issue of some kind. As is often the case, these issues } } may not be related to the reformulation and/or each other. We see every } } one of our customers as very important; we are committed to understanding } } the nature and cause of the difficulty, and finding a way to resolve it. } } Should anyone have a concern or question, please feel free to contact our } } technical support group. } } } } Thank you for your interest in Eastman Kodak Company products. } } } } Respectfully, } } } } Eric Ambrose } } Silver Halide Product Manager } } Scientific Imaging Systems } } Eastman Kodak Company } }
To our knowledge also the Kodak SO163 changed. Does anybody have more information about this? We tried to get information from Kodak about the changed emulsion, but were referred to the sales people ... who did not have the information. The film seems to have gotten less sensitive. I am concerned that this may pose a serious problem for low-dose electron microscopy.
Regards, Michael
__________________________________________ Michael Radermacher University of Vermont College of Medicine Dept. Mol. Physiol. & Biophysics Burlington, VT 05405
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This is Lifeng Dong from Department of Physics, Portland State University, USA. I am planning to attend M&M 2003, and reserved a double room (two beds) from Hampton Inn Downtown San Antonio. The price including tax is $121.42 per night, which is a little expensive for a graduate student. If you or some of your male colleagues will attend this conference and are looking for a hotel, I would like to share it with you or him.
If you have any question, contact me at lifengd-at-pdx.edu.
Post for a friend. -shuyou. ------------------------------------------------------------- Dear All,
I have some troubles on my results. Any comments would be appreciated.
It is about characterization of epitaxial beta-FeSi2. The substrate is Si (111). The thickness of epitaxial Fe film is 10 angstroms. Form beta-FeSi2 islands after annealing at 600C. Bulk beta-FeSi2 belongs to based centered orthorhombic lattice with a=0.9863nm, b=0.7791nm and c=0.7833nm. Epitaxial relationship is (110)beta//(111)Si with [001]beta//[1-10]Si. The fact is that beta-FeSi2 can be viewed as a layered structure based on (110)beta plane, and the atoms on each layer always distribute in the form of distorted hexagon similar to that of Si (111) plane.
Now my question is whether the superimposed diffraction pattern along [111]Si zone axis are same (similar) as the FFT result obtained from the HRTEM images having [111]Si orientation or not.
In my opinion, they are different. The reason is that in real space (110)beta // (111)Si according to the epitaxial relationship. It is clear that [111]Si is perpendicular to (111)Si. But [110]beta is not perpendicular to (110)beta because beta is orthorhombic. On the other hand, the crystal directions should always be perpendicular to the reciprocal planes with the same indices as crystal directions. As a result, in reciprocal space two reciprocal plane (111)*Si and (110)*beta are not parallel each other. The reciprocal plane (111)*Si is perpendicular to electron beam when the orientation is exact [111]Si zone axis, whereas the reciprocal (110)*beta plane is not perpendicular to beam. Another thing is the reciprocal lattice points on (110)*beta plane should distribute in a form of RECTANGLE. So we can see magnified projection of thess rectangles in the superimposed pattern.
We can observe nice plan-view HRTEM images when the orientation is [111]Si. As for FFT of HRTEM image taken from [111]Si orientation, the dots resulting from overlaping beta and Si each other distribute in the form of distorted hexagon. The FFT results show there are two sets of spots, near the Si reflection spot there is always another adjacent spot. Both of them display HEXAGONAL shape in FFT results. I think this is because the atoms distribute in the forms of hexagon in Si and distorted hexagon in beta. In this case both (110)beta and (111)Si planes are perpendicular to beam, but no beta zone axis with simple indices are parallel to beam. The reason why we can see HRTEM images is that there indeed exist some atomic (atom group) planes perpendicular to (110)beta plane although these atomic planes do not have simple indices.
My conclusion is that with same orientation the electron diffraction pattern and FFT of HRTEM image are not always same (similar) if one of them is non-cubic crystal. But somebody tell me this conclusion is stupid wrong. I can not find which step is wrong. Also he did not let me know where is my mistake. Till now I believe it is true. I beg your kind comment for my above conclusion. I am anxious to know any comments. Thank you very much in advance.
Best Wishes,
M. HAN HAN.ming-at-nims.go.jp
_____________________________ Shu-You Li, Ph.D. Electron Microscopist, EPIC NUANCE Northwestern University http://www.nuance.northwestern.edu/EPIC
Having followed several messages on this site, I wanted to know if I already had the "new formulation" in my stock so I asked a few questions and the result of two messages resulted in the following Quotes and the directions that have already been posted about nitrogen bursts/extra agitation with tilting. I also requested that Kodak go back to the old formulation.
"All of the new formulation film is marked on the outside of the container "New Formulation". We started producing the "new formulation" approximately last June. The films expiration date is typically 2 years from the date manufactured. Therefore, you if the film that you have does not specify "new formulation", you may still have the original formulation film."
"The reason for reformulating KODAK Electron Microscope Film 4489 was the direct result of the unavailability of some raw materials necessary in the manufacture of the prior version of this product. Every change to a formulation is accompanied by in-house and customer testing to ensure the product meets its specification. Both in-house and customer testing confirmed that reformulated KODAK Electron Microscope Film 4489 does meet the product specification. Most customers that have used the reformulated KODAK Electron Microscope Film 4489 have found it supplies the same high quality results they have always received from Kodak EM films. However, the various processing techniques performed in the research community can result in some instances where modifications to some protocols are required to achieve comparable results with a reformulated film compared to its predecessor."
This answered my questions and I hope that it will do the same for you.
Remember when Kodak discontinued SO-163 a few decades ago? That was when we switched to 4489 after an extended trial and error period. Or is my mind playing tricks on me?
Pat Connelly Dept. of Biology University of Pennsylvania Philadelphia, PA 19104-6018
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } My question is: why would Kodak substitute this for a perfectly good } film without any warning? The package insert from a box of the old } film was identical to that in the new film, but it seems as if } processing procedures need to be radically altered. This has cost } us some unhappy clients and no end of headaches, not to mention } one embarrassing incident where an employee was blamed for something } that wasn't his fault. } Unless I missed some major press release, this seems like a highly } irresponsible action. Does anyone have any insight into this? } Agitatedly, } Randy } === } Normally I don't like litigation but the pain and suffering and the lack } of warning seems to point to a class action suit, and at least an } official explanation accompanying the new film and properly labeled new } film. I'm maybe just surprised to not have seen that suggestion in the } previous discussion on 4489 formula change. } } I know this has been gone over in length before. We do not have any } nitrogen capabilities in our darkroom, and I am dreading the next batch } of 4489 film. Maybe Kodak is using this like Coca-cola did when they } made "new" coke, and then switched back to "classic" but that change let } them go from using expensive beet sugar to corn sugar as the primary } sweetener saving millions of dollars. Who knows - but from the great } discussion here I at least have a plan for when we get the new formula } (not that I will be able to implement it). } } Geoff Williams } Microscopy Facility Supervisor } CMU Biology Department Microscopy Facility web page. } http://www.cst.cmich.edu/centers/microscopy/
Greetings, In the early and mid 1990's I was the proud owner of a Panasonic Optical Drive, model LF 7010, until it stopped working a few years ago. I now find myself in the embarassing position of wanting to get some data off a disk written by that drive (I could have sworn we backed the stuff up on CD but if so the back up is lost). The Panasonic drive was operated by a Mac, as a scsi device. The disk in question is a 1.5 GB Panasonic Optical Disk (LM-R1500A). I know this is a long shot, but is there anyone out there who might be able to read this disk?? (Panasonic tech support only has PC's nowadays, I checked with their Midwest branch).
It's been my experience that this sort of comment comes from a complete lack of understanding of EM techniques. I have always maintained that it is much, much easier to teach a standard lecture course to 50-100+ students, or a standard lecture & lab course to 20-30 students, than it is to teach an intensive EM course to 8-16 students. The students need much more intensive instruction in lab techniques and equipment operation than in a standard lab course; they usually work at their own convenience (since we don't have an EM or microtome for each student), so they need help throughout the day and week; they usually work on a course project to hone skills, and that requires lots of mentoring and personal problem solving; and let's not forget about the time it takes to keep the equipment running when novices are hard on it. Try counting the time you spend on the course (not limited to in-class time), and compare it to 3 hours of lecture per week. And good luck. --------------------------------------- Jan Robert Factor, Ph.D. Professor of Biology --------------------------------------- Division of Natural Sciences Purchase College State University of New York Purchase, NY 10577 --------------------------------------- Ofc. Tel: 914-251-6659 Ofc. Fax: 914-251-6635 E-mail: jfactor-at-purvid.ns.purchase.edu ---------------------------------------
JERRY KUDENOV 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 Colleagues, } } When teaching an EM course, how many additional non-EM courses might } others teach per semester? I am presently teaching 2 courses, one } of which is on SEM. I also teach microtechnique and a TEM course. } } Obviously, the answer depends on EM course formats and campus size. } } My courses are intensive, usually involving 5-6 students, sometimes } more. An SEM course may take 15-25 hours per week, including all formally } scheduled lab and lecture times, weekly tutorials, solutions preparation } etc.(there is no assistant). Our campus is small, perhaps 14,000 students. } } Since I teach so few students, my Chair is wondering about the fairness } of my workload relative to others in my department who teach many more } students in straight lecture courses (the larger courses have assistants). } } Your answers will be most helpful in clarifying a "normal" academic work } load that includes EM instruction at other similar sized universities. } Please send your replies to me at the e-mail address below. I will } summarize the results and post them on the listserver. } } Thank you in advance! } } Jerry } } --------------------- } Jerry D. Kudenov } Dept. Biological Sciences } University of Alaska Anchorage } 3211 Providence Drive } Anchorage, Alaska 99508 } } afjdk-at-uaa.alaska.edu } Office: 1-907.786.1769 } Fax: 1-907.786.4607 } ----------------------
The annual meeting of the Pacific Northwest Microscopy Society will be held this year at the Pacific Northwest National Laboratory (PNNL) in Richland, WA on May 29 and 30, 2003.
We are soliciting abstracts for contributed and poster presentations in all aspects of light and electron microscopy, as well as in technology advances using microscopy as the analytical technique.
Two parallel sessions will run separately in biological and material science. Biological sciences will focus on applications in fluorescent microscopy including confocal and multi-photon imaging, cathodoluminescence, flow cytometry and immunocytochemistry, and 3-d reconstruction by electron tomography. Contributions to the "technologist's forum" are encouraged. The material science division will have a great Focused Ion Beam (FIB) session; our invited MSA sponsored speaker Dr.Lucille Gianuzzi (University of Central Florida) will give a presentation on this topic.
Abstracts - for both posters and platform presentations should be sent to the program chair, Jim Young (jim.young-at-pnl.gov) in an electronic version (either Word or pdf file), with text and figures not to exceed one page. Deadline for abstract submission is April 30, 2003. Please specify platform or poster session. All accepted papers will be printed (as submitted) in meeting proceedings. We encourage both undergraduate and graduate students to participate in a poster session. Cash prizes of $300, $200 and $100 will be awarded to the best student posters in both biological and material science categories. Vendors - of companies manufacturing or selling products and instrumentation related to microscopy will participate in the exhibit during the meeting. Social event - an evening social including a complimentary catered dinner and wine tasting will be held at the Gordon Brothers winery in Pasco on Thursday evening for all registered participants and spouses. Accommodation - the new PNNL user-housing facility, conveniently located just a short walk from the conference rooms, will be available to the meeting participants at a discounted rate. Rooms with a queen bed and a private bathroom at $50/night includes in room continental breakfast. We have reserved 30 of these rooms at a group rate. You can make a reservation on-line at uhf-at-pnl.gov or call 509-372-6736. Additional accommodation available in Richland hotels. Registration - for the meeting will be free of charge to all current PNMS members (contact Jim Young for membership info). Non-members $10 symposia only, $20 including the Thursday night reception. Badging - for those unfamiliar with the PNNL policies, a visitor badge is required for getting in all PNNL labs. Therefore, each participant wishing to tour the lab facility needs to have a visitor's badge. Conference rooms and the auditorium doesn't require a badge, however we strongly recommend to apply for one in order to participate in in-lab demos and tours. The following information is required: 1. Full name, including middle name 2. Social Security Number 3. Citizenship 4. Date of birth 5. Employer and address 6. Telephone number Additional information is required for foreign nationals, and they should contact Jim Young as soon as possible since the security clearance takes 4-6 weeks.
We are looking forward to see you in Richland!
Alice Dohnalkova PNMS president 2003 PNNL, Richland WA (509)372-0692
Jim Young WA program chair jim.young-at-pnl.gov PNNL, Richland WA (509) 376-2797
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
At 06:26 AM 4/17/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
SO 163 actually has an advantage over 4489 for biological work because it is faster (less problem with moving specimens. which in our lab is THE main problem for biologists). So unless you have a lot of users who routinely enlarge the negative 8 or 10 times....why not try it?
cheers
Sally
Dr Sally Stowe Facility Coordinator, ANU Electron Microscopy Unit Research School of Biological Sciences Australian National University, Canberra ACT0200 AUSTRALIA stowe-at-rsbs.anu.edu.au fax 61 (0)2 6125 3218 http://www.anu.edu.au/EMU
} } } "Patton, David" {David.Patton-at-uwe.ac.uk} 04/17/03 08:45PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
At least with Coke there are other soft drinks. As far as I can tell 4489 is the only film available for routine biological tissue work. I have formed the impression from this listserver that S0163 (?) is for more specialised use. Therefore I think we are over a barrel re voting with our feet (mixed metaphors anyone?).
} ------------------------------------------------------------------------ } 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
} -----------------------------------------------------------------------. } } } Randy, } I'm sure it's got something to do with Kodak saving a fraction of a } penny per film box. Remember Coca-Cola's new formulation years ago? } Seems like Kodak should've taken a lesson. There's a Murphy's Law } corollary which states, " If it ain't broke, don't fix it!" } } WWW } ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } Winston W. Wiggins, Supervisor } Cannon Electron Microscopy Lab } Carolinas Medical Center } P.O. Box 32861; Charlotte, NC 28232-2861 } Ship to: 1000 Blythe Blvd; Charlotte, NC 28203 } Ofc:704-355-1267 ; Lab:704-355-7220 ; Fax 704-355-0589 } E-mail: {mailto:WWiggins-at-Carolinas.org} Winston.Wiggins-at-CarolinasHealthCare.org
} ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ } } } -----Original Message----- } } From: Tindall, Randy D. [SMTP:TindallR-at-missouri.edu] } Sent: Tuesday, April 15, 2003 1:33 PM } To: Clarkson Donna R Contr USAMRD } Cc: microscopy-at-sparc5.microscopy.com } Subject: RE: Kodak 4489 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver
} On-Line Help } {http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html} http://www.msa.microscopy.com/MicroscopyListserver/FAQ.html
} -----------------------------------------------------------------------. } } } Hi, } } We were also taken by surprise by a box of the "New Formulation" 4489 } that we didn't even know we had, until we started using it. This } seems to be the film from hell. We solved the density problem by } increasing our nitrogen burst agitation to 2 seconds every 10 } seconds, but we still are getting strange agitation marks. } } I started out years ago using the method described by Steve Chapman, } which is the Kodak recommendation, and only started using nitrogen } burst when I moved to my present job in 1999. Absolutely no problems } until the new film showed up, then we had very thin negatives with } horrible streaks. As I said, the density problem has been solved, } but we're still fighting the streaks. } } What we are doing is a series of exposure trials using "blank" } exposed negatives, i.e., exposing the films in the scope with no } specimen in place to give an even field of detail-less illumination. } This is guaranteed to show any agitation irregularities. We will } then vary agitation procedures and times until we get acceptable } results. Preliminary results are showing that the plastic film } holders with narrow sides, rather then the ones that reach nearly to } the bottom of the films, cause fewer agitation problems. Other first } results are showing that a combination of "lift and tilt" with } nitrogen burst evens things out. Finally, we were getting a } "thumb-print" on the bottom of the negatives that turned out to be } caused by an agitation swirl around the plastic support rod that } holds the films in place. We solved that one by lifting the films up } slighty so they don't touch the rod, but that only works on holders } that are narrow enough to bend the films when they're inserted. } } When we complete our tests, I hope to post images and procedures on } our website and post a link to the listserver. In the meantime, I'm } following this thread with great interest. } } My question is: why would Kodak substitute this for a perfectly good } film without any warning? The package insert from a box of the old } film was identical to that in the new film, but it seems as if } processing procedures need to be radically altered. This has cost us } some unhappy clients and no end of headaches, not to mention one } embarassing incident where an employee was blamed for something that } wasn't his fault. Unless I missed some major press release, this } seems like a highly irresponsible action. Does anyone have any } insight into this? } } Agitatedly, } Randy } } Randy Tindall } EM Core Facility } University of Missouri, Columbia } } } } } } -----Original Message----- } } From: Clarkson Donna R Contr USAMRD } } [ {mailto:donna.clarkson-at-brooks.af.mil} mailto:donna.clarkson-at-brooks.af.mil]
} Sent: Tue 4/15/2003 8:09 AM } To: bonnie sheppard; Microscopy-at-sparc5.microscopy.com } Cc: } Subject: RE: Kodak 4489 film } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver
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} -----------------------------------------------------------------------. } } } Bonnie, } I have not yet actually used the new 4489 film, but spoke with someone } at Kodak before purchasing it. Seeing all the negative reports on it, I } wanted to "get the scoop" before trying it out. They told me that the best } things to try are additional agitation in both the D-19 and Fixer. If the } negatives still appear uneven or muddy try using a 1:1 dilution of D-19 } instead of the usual 1:2. } Good luck to you and anyone else having to use the new film! I hope I } won't have too much trouble. } Have a good one! } } 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 } } } -----Original Message----- } } From: bonnie sheppard [ {mailto:bls4u-at-cstone.net} mailto:bls4u-at-cstone.net]
} Sent: Monday, April 14, 2003 2:35 PM } To: Microscopy-at-sparc5.microscopy.com } Subject: Kodak 4489 film } } } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } To Subscribe/Unsubscribe -- } {http://www.msa.microscopy.com/MicroscopyListserver} http://www.msa.microscopy.com/MicroscopyListserver
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} -----------------------------------------------------------------------. } } } Is anyone else out there having trouble with "shadows and unevenness" on } 4489 film? We have gone to extensive lengths to be sure our developing } is consistent, and still we are having annoying problems with uneven } (blotchy) negatives. This is not a problem when there is ample tissue } detail in the negative...but it is horrible and a real printiong problem } for folks looking at DNA spreads, where there is little tissue to } negative. } } Help! } } } } } } } } } *********************************************************************** } This electronic message may contain information that is confidential } and/or legally privileged. It is intended only for the use of the } individual(s) and entity named as recipients in the message. If you } are not an intended recipient of this message, please notify the } sender immediately and delete the material from any computer. Do not } deliver, distribute or copy this message, and do not disclose its } contents or take any action in reliance on the information it } contains. Thank you. }
---------------------------------------- Patton, David Email: David.Patton-at-uwe.ac.uk "University of the West of England"
On Thursday, April 17, 2003, at 03:45 AM, Patton, David wrote:
} I have formed the impression from } this listserver that S0163 (?) is for more specialised use.
Dear David, SO163 is more sensitive than 4489, and it can be push-processed to double its sensitivity. The grain size is small enough that the negatives can be scanned with a small pixel size, say 5 micrometers, and get acceptable quantitation. These features make it better for low-dose EM than 4489. Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
I have some troubles on my results. Any comments would be appreciated.
It is about characterization of epitaxial beta-FeSi2. The substrate is Si (111). The thickness of deposited Fe film is 10 angstroms. Form beta-FeSi2 islands after annealing. Bulk beta-FeSi2 belongs to based centered orthorhombic lattice with a=0.9863nm, b=0.7791nm and c=0.7833nm. Epitaxial relationship is (110)beta//(111)Si with [001]beta//[1-10]Si. The fact is that beta-FeSi2 can be viewed as a layered structure based on (110)beta plane, and the atoms on each layer always distribute in the form of distorted hexagons similar to that of Si (111) plane.
Now my question is whether the superimposed diffraction pattern along [111]Si zone axis are same (similar) as the FFT result obtained from the HRTEM images having [111]Si orientation or not.
In my opinion, they are different. The reason is that in real space (110)beta // (111)Si according to the epitaxial relationship. It is clear that [111]Si is perpendicular to (111)Si. But [110]beta is not perpendicular to (110)beta because beta is orthorhombic. On the other hand, the crystal directions should always be perpendicular to the reciprocal planes with the same indices as crystal directions. As a result, in reciprocal space two reciprocal plane (111)*Si and (110)*beta are not parallel each other. The reciprocal plane (111)*Si is perpendicular to electron beam when the orientation is exact [111]Si zone axis, whereas the reciprocal (110)*beta plane is not perpendicular to beam. Another thing is the reciprocal lattice points on (110)*beta plane should distribute in a form of RECTANGLE. So we can see the magnified projection of thess rectangles in the superimposed pattern.
We can also observe nice plan-view HRTEM images when the orientation is [111]Si. As for FFT of HRTEM image taken from [111]Si orientation, the dots resulting from overlaping beta and Si each other distribute in the form of distorted hexagon. The FFT results show there are two sets of spots, near the Si reflection spot there is always another adjacent spot. Both of them display HEXAGONAL shape in FFT results. I think this is because the atoms distribute in the forms of hexagon in Si and distorted hexagon in beta. In this case both (110)beta and (111)Si planes in real space are perpendicular to beam, but no beta zone axis with simple indices are parallel to beam. The reason why we can see HRTEM images in this case is that there indeed exist some atomic (atom group) planes perpendicular to (110)beta plane although these atomic planes do not have simple indices.
My conclusion is that with same orientation the electron diffraction pattern and FFT of HRTEM image are not always same (similar) if one of them is non-cubic crystal. But somebody tell me this conclusion is stupid wrong. I can not find which step is wrong. Also he did not let me know where is my mistake. Till now I believe it is true. I beg your kind comment for my above conclusion. I am anxious to know any comments. Thank you very much in advance.
Below is the result of your feedback form (NJZFM-ultra-55). It was submitted by (cperalta-at-wisc.edu) from http://www.msa.microscopy.com/Ask-A-Microscopist.html on Friday, April 18, 2003 at 08:18:09 ---------------------------------------------------------------------------
Email: cperalta-at-wisc.edu Name: Carlos Peralta
Organization: University of Wisconsin
Education: Graduate College
Location: Madison, Wisconsin, USA
Question: Hello: I am getting ready to mount conodonts on glass slides. I'll be using gum tragacanth. Can you tell me what compound should I add to prevent fungal growth. Any answer or suggested sources will be greatly appreciated.
I already asked my colleagues but I didn't get a good answer.
Thank you to everyone who responded to my question regarding the best oil mist filtration system for venting my vacuum pumps indoors.
I received several responses that boiled down to about three basic ideas.
1) Using a favorite brand name (their were several) filter on each pump.
2) Connecting one very large brand name filter to all of the pumps.
3) Venting to the outside air with or without some kind of filter.
Someone also suggested using modified generic automobile engine oil filters.
Regards, Jim
James S. Romanow The University of Connecticut Physiology and Neurobiology Department Electron Microscopy Facility Unit 2242 354 Mansfield Road Beach Hall, Room 129 Storrs, CT 06269-2242
Pat, I do not know where you got your information but Kodak has been making SO-163 for years and is still doing so. We have used it since the late 80's and have not had any problem with developing (or new formulations so far!). It gives excellent images. Those of you having problems with 4489 might want to try this film. 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
On 4/17/03 2:02 PM, "Pat Connelly" {psconnel-at-sas.upenn.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 } -----------------------------------------------------------------------. } } } Having followed several messages on this site, I wanted to know if I } already had the "new formulation" in my stock so I asked a few } questions and the result of two messages resulted in the following } Quotes and the directions that have already been posted about } nitrogen bursts/extra agitation with tilting. I also requested that } Kodak go back to the old formulation. } } "All of the new formulation film is marked on the outside of the } container "New Formulation". We started producing the "new formulation" } approximately last June. The films expiration date is typically 2 } years from the date manufactured. Therefore, you if the film that you } have does not specify "new formulation", you may still have the original } formulation film." } } "The reason for reformulating KODAK Electron Microscope Film 4489 was the } direct result of the unavailability of some raw materials necessary in } the manufacture of the prior version of this product. Every change to a } formulation is accompanied by in-house and customer testing to ensure } the product meets its specification. Both in-house and customer testing } confirmed that reformulated KODAK Electron Microscope Film 4489 does } meet the product specification. Most customers that have used the } reformulated KODAK Electron Microscope Film 4489 have found it supplies } the same high quality results they have always received from Kodak EM } films. However, the various processing techniques performed in the } research community can result in some instances where modifications to } some protocols are required to achieve comparable results with a } reformulated film compared to its predecessor." } } This answered my questions and I hope that it will do the same for you. } } Remember when Kodak discontinued SO-163 a few decades ago? That was } when we switched to 4489 after an extended trial and error period. Or } is my mind playing tricks on me? } } Pat Connelly } Dept. of Biology } University of Pennsylvania } Philadelphia, PA 19104-6018 } } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } } } My question is: why would Kodak substitute this for a perfectly good } } film without any warning? The package insert from a box of the old } } film was identical to that in the new film, but it seems as if } } processing procedures need to be radically altered. This has cost } } us some unhappy clients and no end of headaches, not to mention } } one embarrassing incident where an employee was blamed for something } } that wasn't his fault. } } Unless I missed some major press release, this seems like a highly } } irresponsible action. Does anyone have any insight into this? } } Agitatedly, } } Randy } } === } } Normally I don't like litigation but the pain and suffering and the lack } } of warning seems to point to a class action suit, and at least an } } official explanation accompanying the new film and properly labeled new } } film. I'm maybe just surprised to not have seen that suggestion in the } } previous discussion on 4489 formula change. } } } } I know this has been gone over in length before. We do not have any } } nitrogen capabilities in our darkroom, and I am dreading the next batch } } of 4489 film. Maybe Kodak is using this like Coca-cola did when they } } made "new" coke, and then switched back to "classic" but that change let } } them go from using expensive beet sugar to corn sugar as the primary } } sweetener saving millions of dollars. Who knows - but from the great } } discussion here I at least have a plan for when we get the new formula } } (not that I will be able to implement it). } } } } Geoff Williams } } Microscopy Facility Supervisor } } CMU Biology Department Microscopy Facility web page. } } http://www.cst.cmich.edu/centers/microscopy/ } } }
Does anyone out there have an Amray SEM not being used that has the External Beam Interface plate? I would appreciate getting one of these plates. Will pay packing and shipping.
It is a small aluminum plate inside the rear of the main console. There are two BNC connectors ( X Y ) and a screw terminal strip. coming out of the plate is a cable with a 6-pin AMP connector that says P9 on it.
I think this needs a bit clarification. More is not always better.
The theoretical requirements for optimum sample frequency were calculated by Shannon and Nyquist some time ago. They basically arrive at the same results: To measure or transmit a frequency X without artifacts, you need to sample the signal with frequency 2X. Staying below this frequency will introduce artifacts, going above it will not yield much improvement.
In other words, if your microscope is capable of resolving 1 micron, you need to use a camera that can resolve 0.5 microns. To find out the matching magnification on the microscope (b/w camera), you solve PS/mag=0.5, where PS is the Pixel size of the camera, and mag is the total magnification between object and camera chip (note that there may be other lenses in the beam path).
For color cameras the situation is more complex, as the resolution is actually color dependend. A Bayer-type color chip has twice as many green pixels as red and blue. For green object, therefore, the resolution is half that of the chip, for red and blue one quarter. If you have a grey object, you'd get full resolution as all pixels are sensitive to some of the grey spectrum.
As for the printing mentioned below: If you are limited by the instrument resolution, and you are using a camera with "too many" pixels, all you get is empty resolution, which is the same as using a camera with a lower resolution (fewer pixels) and artificially increasing the number of pixels and interpolate. The same is true for posters or large prints: Whether you "grossly oversample" the image by using a camera with way too many pixels or artificially increase the image size with interpolation doesn't matter. All you get is empty resolution.
Finally, the "resolution improvement". Image processing is no magic bullet. I have seen those scenes on TV or in Movies, where somebody takes a blurry picture of a crowd, and someone takes a single face, which now looks like a grey blob, then "sharpens" the image to a point where you can see the stubbles of the man's five 'o clock shadow. That's fiction. There are some ways of improving resolution by using deconvolution techniques, but they usually require additional information about the measurement setup (modulation transfer functions, etc). Most other image processing functions make certain information in the image more visible (or less visible), but don't actually improve the resolution. For example a "sharpen" filter does not add information to the image. It does make contrast transitions more visible and thus creates the impression of a "sharper" image, but it also increases noise.
So much for my 2 cents worth...
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Frank Eggert [mailto:Eggert-at-mikroanalytik.de] Sent: Thursday, April 17, 2003 1:41 AM To: Gary Radice Cc: Microscopy-at-sparc5.microscopy.com
Gary,
you are right. From the point of microscopy there are only two reasons to have more pixels:
#1 To print of large images (e.g. for posters or for other representations) without loss of details. #2 For digital processing of your images (e.g. to 'improve' resolution of instrument or to extract details).
Or are there other reasons?
} From a major interest should be the data depth (possible grey steps in each pixel) for a computer processing of brightness and contrast. If you have high dynamics, even wrong lighted images are able to convert into better results or you are able to improve contrast of low differences in an image region, which is uniform in colour and brightness with first views.
Frank Eggert
http://www.microanalyst.net
Gary Radice schrieb:
} ------------------------------------------------------------------------ } 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 the option to buy either a 5 or 3 megapixel camera. What I want } to know is whether a 5 megapixel camera is better than a 3 megapixel } camera, if used exclusively on a microscope, or whether the extra } pixels (and cost) are wasted. Assume bit depth is the same. } } I'd like someone to check my math and my reasoning. } } I understand that microscope resolution is limited by diffraction and a } good rule of thumb for displaying images at normal reading distance is } a final image mag of no more than 1000x the numerical aperture of the } objective. On my microscope this means about 100x for images taken with } the 4x lens and 1250x for images with the 100x oil lens. Assuming I use } a 1x relay eyepiece to connect the CCD camera, the image at the } detector could be magnified 25 times for the 4x objective, and 12.5 } times for the 100x objective, before getting into empty mag. } } On the camera side, I understand that the more pixels the better the } resolution. The 3 megapixel camera has 2080 horizontal pixels and the } 5mp has 2580. Let's ignore the vertical pixels for the moment. } } One more assumption is that for printing, it seems that the highest } practical output resolution is 266-300 dpi. This means that the 3 mp } camera produces an image that optimally prints an image 2080/266 = 7.8 } inches or 198 mm horizontally, or less, and the 5 mp can print an image } about 243 mm horizontally or less. Enlarging above these dimensions is } possible but the print image will need interpolation and might be } degraded. } } Going back to the microscope, the maximum horizontal field of view with } the 4x objective captured by the camera is 1.6 mm. It could be printed } up to 100 x 1.6 = 160 mm before empty mag sets in. But I have already } calculated that the 3 mp can print up to 198 mm and the 5 mp up to 243 } mm before the print image degrades. } } If these calculations are correct (please tell me if it they are not) } then both the 3 mp and 5 mp have more than enough resolution to capture } what the microscope makes available at 4x, and the situation is even } better for the 100x lens because the lens does most of the heavy } lifting, magnification-wise. } } One final condition that might matter is that both are RBG cameras with } Bayer filters, so that the real resolution is approximately 1/3 of the } nominal pixel resolution. I'm not sure how to figure that into the } calculations. } } So, at approximately a dollar-per-pixel, do I need the extra 2000 } pixels of a 5 mp camera? } } Gary P. Radice gradice-at-richmond.edu } Department of Biology 804-289-8107 (voice) } University of Richmond 804-289-8233 (FAX) } Richmond VA 23173 http://www.richmond.edu/~gradice
Debby, When I started at U of P in 1976, the film in use was the SO-163 and it was in use until either the late 70's or early 80's. I took off three years, had two children and when I returned, the film had been switched to Kodak 4489. I believe that so many researchers complained, that Kodak re-issued the film but we decided not to switch back from the 4489 that we were, at that time, using as the replacement. I am considering the switch back to the SO-163 now, in that by using either it or the "new formulation 4489" I will have to adjust my protocol and scope (Philips 200) settings. (There are no nitrogen burst tanks here.)
Can anyone tell me when using the new formulation 4489, if the negatives are darker or lighter than the old formulation before making adjustments to exposure time or sensitivity or is the total problem just the proper agitation of the developer?
I am blessed to have ordered, by chance, a quantity of film while the old supply still was available.
Thanks, Pat Connelly
} Pat, } I do not know where you got your information but Kodak has been making } SO-163 for years and is still doing so. We have used it since the late } 80's and have not had any problem with developing (or new formulations so } far!). } It gives excellent images. Those of you having problems with 4489 might } want to try this film. } 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 } } On 4/17/03 2:02 PM, "Pat Connelly" {psconnel-at-sas.upenn.edu} wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America } } -----------------------------------------------------------------------. } } Remember when Kodak discontinued SO-163 a few decades ago? That was } } when we switched to 4489 after an extended trial and error period. Or } } is my mind playing tricks on me? } } Pat Connelly } } Dept. of Biology } } University of Pennsylvania } } Philadelphia, PA 19104-6018
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
This is exactly the test I performed (we alternated old/new sheets) with exactly the same density results, when photographing a "typical" biological sample. I used freshly made Kodak D-19 developer diluted 1:2 (as recommended by Kodak) and Kodak Rapid Fixer; the temperature was within 1/2 degree of the recommended 20 degrees C, using constant agitation, occasionally lifting the negatives completely out of the developer, tilting first to one side then the next to get the developer in the grooves of the Lucite holder. All of the negatives were evenly developed, but the new formulation was significantly darker than the old formulation. I have not yet adjusted the exposure (sensitivity) of the microscope to the film since we still have a little of the old film left, but I expect that this should be much easier (and cheaper!) than trying to implement a nitrogen bust system, which we've never had nor needed despite Kodak's instructions.
I've been following this discussion since early February when I first posted a question myself about the film, and was rather surprised to see that my test result was quite different from everything I had read. Glad to see that someone's results finally corroborate mine!
Valerie
At 09:10 AM 4/17/2003 +0200, R. Cross wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Valerie M. Knowlton Research Assistant/Teaching Technician Center for Electron Microscopy 1219 Gardner Hall, Box 7615 North Carolina State University Raleigh, NC 27695
I have a user who wants to make TEM carbon platinum replicas of mammalian cells growing on glass slides with attached tissue tek culture chambers. He figures to fix the cells, dehydrate, critical point dry, then coat the slides (minus the growth chambers) and float the replicas off the slide the same way he floats formvar off a slide when making coated grids. In his first attempt, he is unable to float the metal coating off the slide. A query to this listserver quite a while back mentioned using hydrofluoric acid to remove the replica. does anyone have details of this procedure, or preferably an alternative procedure that will enable my user to make his replicas?
Thanks for any insights in this regard
Steve -- Dr. Steven Barlow EM Facility/Biology Dept. San Diego State University 5500 Campanile Drive San Diego CA 92182-4614 phone: (619) 594-4523 fax: (619) 594-5676 email: sbarlow-at-sunstroke.sdsu.edu http://www.sci.sdsu.edu/emfacility
Chairman, Educational Outreach subcommittee promoting microscopy instruction and increased access to microscopes Microscopy Society of America http://www.msa.microscopy.com/
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
A friend from our Soil Science department wants to compare roots from live plants grown in good soil and dead plants grown in poisoned soil. So far we have been looking at them under dark-field optical microscopy. It would be good if we could look at them under SEM, but we don't have an ESEM, and we don't want the root shape to collapse under vacuum dessication.
When I have searched for root fixation I come across things like glutaraldehyde and cacodylate buffer, and we'd like to avoid using these if possible. Does anyone know simpler ways of fixing the external shape of roots?
It would be nice if we could do the same thing for moulds, to show visiting students "this is what you get if you leave bread lying around in your accommodation!".
Any ideas, please?
+-----------------------------------------+ 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 +----------------------------------------+
Dear Steve If it's Pt or Pt/C coating to remove replica from the glass you may use some HF solution. I don't remember exact concentration, but I would think, 10% (v/v) should work. If not, you may try higher concentration. Please, keep in mind: HF is extremely aggressive chemical: it's strong irritant and will corrode everything on it's way including SS EM tweezers. It should be stored in the plastic (NO GLASS) and used with precautions. Basically, you use the same technique as for floating Formvar film except everything should be plastic and HF solution instead of water. When replica is detached from the glass, you may move it with Pt loop to the water to wash it a few times and then mount on the grids. Good luck. Sergey.
At 12:55 PM 4/21/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Dear Ming, This problem is in principle quite simple. Any diffraction pattern can be seen as a Fourier transform of the diffracting object. You can find this in any undergraduate optics textbook. Normally in TEM you will then have to convulute the periodicity of the periodicity of the object with a top- hat function describing its very limited thickness. This leads to the well-known reciprocal lattice rods for thin samples.
So, an SAD pattern is a Fourier transform including specimen periodicity and specimen thickness.
An HRTEM image can also be Fourier transformed. As you should know, however, the HRTEM image is strongly influenced by the contrast transfer function (i.e. highly defocus dependent). That means that the intensities in a Fourier-transform of a HRTEM image are strongly affected by image defocus. You see this effect particularly strongly for amorphous material.
So, an SAD pattern and a Fourier-transformed HRTEM image have the same geometry, but may have rather different intensities in the different spots since the latter is so focus-dependent.
That said, in your case the main point is that the geometry of the pattern is identical for SAD or for Fourier-transformed HRTEM.
Now, if your orientation relationship is correct, then when you orient along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are matching the (111) Si planes. I reckon on a quick calculation that the angle between the (110)beta plane normal and [110]beta is 13.4 °, much too large to see the [110]beta diffraction pattern. But the axis [230]beta lies only 1.86° from the (110)beta plane normal. Are you seeing this diffraction pattern? At such a small angle off axis, you will still see the diffraction pattern and an HRTEM image (even if it’s not very good or symmetric).
Best wishes
Ian MacLaren
------- Forwarded message ------- } From: HAN.Ming-at-momokusa.nims.go.jp To: Microscopy-at-sparc5.microscopy.com
ATTN:
} From the Desk of MR MIKE OBI AFRI Bank of Nigeria, Lagos-Nigeria.
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Greetings, Fixation will distort the organ, no matter what. But you should still be able to compare your samples. Aldehydes give the best fixation, as a rule, but there is no reason to use cacodylate for your purpose (actually, some would say no reason ever, but that is another story). Plant roots can be fixed in FAA, dehydrated to absolute ethanol, critically point dried, coated with metal, and viewed. As an alternative to FAA, you could use 4% paraformaldehyde in say PBS (or the dilute buffer of your choice, near pH7), rinse in the buffer, dehydrate etc as above. I expect that 2 h in 4% paraformaldehyde fix would be fine. You can check for shrinkage and other untoward affects by looking at your fixed or dehydrated roots under darkfield.
Hope this helps, Tobias } } } } A friend from our Soil Science department wants to compare roots from live } plants grown in good soil and dead plants grown in poisoned soil. So far } we have been looking at them under dark-field optical microscopy. It } would be good if we could look at them under SEM, but we don't have an } ESEM, and we don't want the root shape to collapse under vacuum } dessication. } } When I have searched for root fixation I come across things like } glutaraldehyde and cacodylate buffer, and we'd like to avoid using these } if possible. Does anyone know simpler ways of fixing the external shape } of roots? } } It would be nice if we could do the same thing for moulds, to show } visiting students "this is what you get if you leave bread lying around in } your accommodation!". } } Any ideas, please? } } +-----------------------------------------+ } 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 } +----------------------------------------+
I´m interested in labelling bacterial surfaces with colloidal gold for negative stain and TEM. So far negative stain works OK, also indirect immunofluorescence with a primary mouse monoclonal. However, secondary antibodies conjugated with 10-15 nm gold doesn´t work at all. We have adhered fimbriated E Coli to Formvar-coated gold grids with or without fixation in various concentrations of formaldehyde, labelled them and fixed with glutaraldehyde. Any tips and tricks?? Regards, Margaretha
--
Margaretha Lindroth, Ph D Phone: 4613-222616 Dept of Medical Microbiology Fax: 4613-224789 Faculty of Health Sciences e-mail: Margaretha.Lindroth-at-hul.liu.se Linköpings Universitet SE-581 85 Linköping
I would greatly appreciate your reponse. I believe you spent much time answering my stupid question.
In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this sphere is not a plane. Sometimes it brings some troubles to us. Let me show an example. If the specimen normal is NOT parallel to incident beam, or if the orientation is NOT exact, the intersections between relrod and sphere are not symmetric. In SAD pattern, not only intensity but also the distance is different, in other words, SAD pattern is NOT symmetric. On the other hand, FFT always symetric even though zone axis is not exact. This let me believe FFT and SAD is not so identical.
Thanks for your kind calculation. Near [111]Si orientation, we can find SAD pattern. As you said, it is not an exact zone axis. I took three patterns with different tilting angles (x=-9, +2, +8) and spent two weeks understanding these patterns. After calibrating the tilting angles by Kikuchi pattens, my indexing results let me believe they come from an identical reciprocal plane (130)*. I believe your calculation is right. I guess the reasons why it is [130] rather than [230] maybe come from two points. 1. beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. (640) spot is too far from transmitted spot. 2. The epitaxial phase is under distorted state. The real angle maybe different from those calculated based on bulk data.
Till now I still think FFT is not completely identical to SAD. I believe the discussion itself is significant. In my opinion, we can view Ewald sphere as a plane and based on this approximation we can think FFT=SAD. But for epitaxy, we can not ignore this approximation if we want to discuss some slight mismatch information. Maybe I made some mistakes in my analysis. I hope to hear different sounds and look forward to hearing from you again.
Thanks again,
Best wishes,
Han
} Dear Ming, } This problem is in principle quite simple. Any diffraction pattern can be } seen as a Fourier transform of the diffracting object. You can find this } in any undergraduate optics textbook. Normally in TEM you will then have } to convulute the periodicity of the periodicity of the object with a top- } hat function describing its very limited thickness. This leads to the } well-known reciprocal lattice rods for thin samples. } } So, an SAD pattern is a Fourier transform including specimen periodicity } and specimen thickness. } } An HRTEM image can also be Fourier transformed. As you should know, } however, the HRTEM image is strongly influenced by the contrast transfer } function (i.e. highly defocus dependent). That means that the intensities } in a Fourier-transform of a HRTEM image are strongly affected by image } defocus. You see this effect particularly strongly for amorphous material. } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } geometry, but may have rather different intensities in the different spots } since the latter is so focus-dependent. } } That said, in your case the main point is that the geometry of the pattern } is identical for SAD or for Fourier-transformed HRTEM. } } Now, if your orientation relationship is correct, then when you orient } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } matching the (111) Si planes. I reckon on a quick calculation that the } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much too } large to see the [110]beta diffraction pattern. But the axis [230]beta } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } diffraction pattern? At such a small angle off axis, you will still see } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } symmetric). } } Best wishes } } Ian MacLaren }
The Pt-C replicas can be detached from the glass substrate by submerging the specimen at a shallow angle (} 45º) into a small petri dish containing 6 to 10% HF. If the concentration is too high, the replica may disintegrate into small pieces. To clean the replicas of any biological material adhering to the replica undersurface, you may have to transfer to 70% sulphuric acid. Another choice would be 1- 2 M NaOH. After a few hours, the replicas should be transferred to distilled water and then to 400 mesh TEM grids. Any remaining water should be removed with filter paper. A good technical reference is Zeile, U. (2000) Fundamentals of cryo preparation and replica technique. 7th Asia-Pacific Electron Microscopy Conference. Reprints are available from Bal-Tac. However, other methods to avoid the creation of artefacts in biological specimens is to use an Environmetal SEM (LV-SEM, VP-SEM) or the cryo method for rapid freezing (plunge freezing, jet-freezing, or high pressure freezing) and the use a freeze-fracture system for Pt-C replicas.
-- Kelly
} } Dear Steve } If it's Pt or Pt/C coating to remove replica from the glass you may } use some HF solution. I don't remember exact concentration, but I } would think, 10% (v/v) should work. If not, you may try higher } concentration. Please, keep in mind: HF is extremely aggressive } chemical: it's strong irritant and will corrode everything on it's way } including SS EM tweezers. It should be stored in the plastic (NO } GLASS) and used with precautions. Basically, you use the same } technique as for floating Formvar film except everything should be } plastic and HF solution instead of water. When replica is detached } from the glass, you may move it with Pt loop to the water to wash it a } few times and then mount on the grids. Good luck. Sergey. } } At 12:55 PM 4/21/2003, you wrote: } } } } } } } Dear all } } } } I have a user who wants to make TEM carbon platinum replicas of } } mammalian cells growing on glass slides with attached tissue tek } } culture chambers. He figures to fix the cells, dehydrate, critical } } point dry, then coat the slides (minus the growth chambers) and float } } the replicas off the slide the same way he floats formvar off a slide } } when making coated grids. In his first attempt, he is unable to } } float the metal coating off the slide. A query to this listserver } } quite a while back mentioned using hydrofluoric acid to remove the } } replica. does anyone have details of this procedure, or preferably } } an alternative procedure that will enable my user to make his replicas? } } } } Thanks for any insights in this regard }
Mike; Yes, Nyquist was an optimist. The 44 hz frequency rate for CDs is for stereo, and the 22hz sampling rate per channel has been abandoned in the newer SACD standard which oversamples 8 times (that's 176 kz).
John Mardinly
John Mardinly Intel
-----Original Message----- } From: Mike Bode [mailto:mb-at-Soft-Imaging.com] Sent: Monday, April 21, 2003 1:11 PM To: 'Microscopy-at-MSA.Microscopy.Com'
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
} } FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD.
I'm afraid not. In fact, the FFT (of the image) is NEVER the same as the SAD. This is a point that I have always tried to stress when teaching at the NCEM Summer Schools.
The FFT (or image diffractogram) is always complex hermitian because it is the FT of a real function (the image intensity) -- that means that the intensities of the FFT (or image diffractogram) are always symmetric (as you point out) for ANY image at ANY specimen tilt or ANY misorientation.
For a crystalline specimen, producing strong Bragg spots in the diffraction pattern, the spots in the FFT (or image diffractogram) are well-defined combinations of interferences of the Bragg spots in the diffraction pattern. These interferences come from the auto-correlation (a kind of convolution) of the (complex) Bragg spot amplitudes (after the Bragg spot amplitudes have been modulated by the phase changes imposed by the objective lens). For a brief discussion of how the spots in the FFT (or image diffractogram, or image intensity spectrum) are formed from the Bragg spots in the diffraction pattern, see "Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, San Antonio, Texas (1979) 556-557.
Of course, none of the above helps you with your orientation problem -- sorry.
Mike O'Keefe
$B4Z(J $BL-at-(J 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 Ian MacLaren, } } I would greatly appreciate your reponse. I believe you spent much time answering my } stupid question. } } In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is true if Ewald } sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this sphere is not } a plane. Sometimes it brings some troubles to us. Let me show an example. If the specimen } normal is NOT parallel to incident beam, or if the orientation is NOT exact, the } intersections between relrod and sphere are not symmetric. In SAD pattern, not only } intensity but also the distance is different, in other words, SAD pattern is NOT } symmetric. On the other hand, FFT always symetric even though zone axis is not exact. } This let me believe FFT and SAD is not so identical. } } Thanks for your kind calculation. Near [111]Si orientation, we can find SAD pattern. As } you said, it is not an exact zone axis. I took three patterns with different tilting } angles (x=-9, +2, +8) and spent two weeks understanding these patterns. After } calibrating the tilting angles by Kikuchi pattens, my indexing results let me believe } they come from an identical reciprocal plane (130)*. I believe your calculation is right. } I guess the reasons why it is [130] rather than [230] maybe come from two points. 1. } beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. (640) spot is } too far from transmitted spot. 2. The epitaxial phase is under distorted state. The real } angle maybe different from those calculated based on bulk data. } } Till now I still think FFT is not completely identical to SAD. I believe the discussion } itself is significant. In my opinion, we can view Ewald sphere as a plane and based on } this approximation we can think FFT=SAD. But for epitaxy, we can not ignore this } approximation if we want to discuss some slight mismatch information. Maybe I made some } mistakes in my analysis. I hope to hear different sounds and look forward to hearing } from you again. } } Thanks again, } } Best wishes, } } Han } } } Dear Ming, } } This problem is in principle quite simple. Any diffraction pattern can be } } seen as a Fourier transform of the diffracting object. You can find this } } in any undergraduate optics textbook. Normally in TEM you will then have } } to convulute the periodicity of the periodicity of the object with a top- } } hat function describing its very limited thickness. This leads to the } } well-known reciprocal lattice rods for thin samples. } } } } So, an SAD pattern is a Fourier transform including specimen periodicity } } and specimen thickness. } } } } An HRTEM image can also be Fourier transformed. As you should know, } } however, the HRTEM image is strongly influenced by the contrast transfer } } function (i.e. highly defocus dependent). That means that the intensities } } in a Fourier-transform of a HRTEM image are strongly affected by image } } defocus. You see this effect particularly strongly for amorphous material. } } } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } } geometry, but may have rather different intensities in the different spots } } since the latter is so focus-dependent. } } } } That said, in your case the main point is that the geometry of the pattern } } is identical for SAD or for Fourier-transformed HRTEM. } } } } Now, if your orientation relationship is correct, then when you orient } } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } } matching the (111) Si planes. I reckon on a quick calculation that the } } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much too } } large to see the [110]beta diffraction pattern. But the axis [230]beta } } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } } diffraction pattern? At such a small angle off axis, you will still see } } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } } symmetric). } } } } Best wishes } } } } Ian MacLaren } }
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SACD was launched in 1999 and is based on a different technology for the CDs (you won't be able to play the SAcDs on your regular CD player). I looked this up because, frankly, I had not even heard of SACD. They also claim they can reproduce sound up to 100 kHz (on the Sony SACD site). That's even too high for my dog, perhaps bats can hear that. And what speaker can reproduce that? But again: 176 kHz sampling rate for a 100 kHz signal is again roughly a factor of 2.
What they are saying is, that the reproduction of higher frequencies can produce better sound. That may well be. In order to sample the higher frequencies, they have to go to higher sampling rates, and they end up again at roughly Nyquist.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, April 22, 2003 10:42 AM To: Mike Bode; Microscopy-at-MSA.Microscopy.Com
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
I can't help Tobias, but if anybody out there has a need for a Panasonic LF-5090 (sold through Corel) I can let them have it for the cost of shipping.
This unit writes to LM-D501W disks (940 MB double sided - 470 MB per side). I don't have any spare media, though. I know it works as I lashed it together over the Christmas holidays to transfer all of my files from it to CD. I've never been able to get it to work under Windows, but it works under DOS on my old, original Compaq Portable just fine (my age is showing). I've got the ISA SCSI card that it seems to like, all the documentation, and several (I think) versions of the software (available at www.driverguide.com as well). It's just taking up space here, so if anybody needs it to keep an old piece of equipment viable, let me know.
Cheers,
Jim
--
James M. Ehrman Digital Microscopy Facility Mount Allison University Sackville, NB E4L 1G7 CANADA
} Greetings, } In the early and mid 1990's I was the proud owner of a } Panasonic Optical Drive, model LF 7010, until it stopped working a } few years ago. I now find myself in the embarassing position of } wanting to get some data off a disk written by that drive (I could } have sworn we backed the stuff up on CD but if so the back up is } lost). The Panasonic drive was operated by a Mac, as a scsi device. } The disk in question is a 1.5 GB Panasonic Optical Disk (LM-R1500A). } I know this is a long shot, but is there anyone out there who might } be able to read this disk?? (Panasonic tech support only has PC's } nowadays, I checked with their Midwest branch). } } Thanks, } Tobias
Mike, you are missing the point: as pointed out by Professor Marks, just Nyquist just doesn't quite cut it.
John Mardinly Intel
-----Original Message----- } From: Mike Bode [mailto:mb-at-soft-imaging.com] Sent: Tuesday, April 22, 2003 10:41 AM To: 'Microscopy-at-MSA.Microscopy.Com' Cc: Mardinly, John
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
Several weeks ago, I asked on references of dose measurement in microscope. I am still looking for that. This time, I want to more specify my question. I would appreciate any comments you have.
Here, I simply consider current density instead of dose. There is a relation between current densities on specimen (Is) and Faraday cup (If).
Is = If x M x M
Where M is a magnification on the Faraday cup.
The relation between magnifications on Faraday cup (M) and film (m) are given by m/M = l/L where L indicates the distance between Projection Lens and Faraday cup, and l indicates the distance between Projection Lens and film. We know magnification on film (m) and can measure the current density on Faraday cup, so we can calculate the current density on the specimen.
I am looking for a textbook or paper that described the method I wrote above. Please advise.
Thank you,
Hiromi Konishi, Ph.D Dept. of Earth and Planetary Sciences The University of New Mexico
i am sure you will get dozens of responses, all with good advice. the only thing i would mention is some early work we did with chlamydial elementary bodies. the problem was that the monoclonal antibody worked very well with the western blot and, supposedly with immunofluorescent microscopy. i tried making my own gold and labelling it directly with the antibody, using protein A as an interface with the gold and antibody, and indirect, with purchased gold anti-mouse. none of them worked. my only explaination is, to this day, that the problem with monoclonal antibodies is accessability and conformation of their target site. if it is inside the cell, or buried in a hydrophobic fold or globular structure, the antibody will not work. it sounds as if one answer is that your antigenic target is not accessable. for what it is worth, i have had good success with each of the gold labelling methods, direct Ab to gold, using protein A, or using store bought stuff. and the store bought stuff is soooooo much easier because you do not have to make it yourself.
i would suggest that you try repeating the work with polyclonal antisera. i have always had much better success with them. the chances of an antibody which will recognize an accessable site is much greater.
paul hazelton, PhD EM Unit Medical Microbiology and Infectious Diseases University of Manitoba Winnipeg, Manitoba, Canada phone 204-789-3313 fax: 204-789-3926 pager: 204-931-9354
Ok, John, maybe I AM not getting the point, but here is as I see it:
1) Both Nyquist and Shannon calculated the THEORETICAL sampling frequency to measure a frequency without introducing artifacts. They both arrive at 2X. In a practical setting (camera with noise), the frequency may well be higher. How much a few percent noise will add to the sampling frequency I don't know, but my guess is, it's not a lot.
2) You mentioned the new SACDs. They use a higher sampling frequency to be able to reproduce higher acoustic frequencies because the claim is, that the music sounds better then (which I can't make any statement about). However, their own web site claims, that they can reproduce up to 100 kHz. With a 176 kHz sampling frequency, they are staying BELOW Nyquist and claim they can reproduce 100 kHz sound. I am not saying that they can or can't or that it sounds better or worse, just an observation that the guys at Sony and Philips use numbers similar to Nyquist.
Maybe we should move this discussion off-line before we start boring people.
I'd be interested to hear if you have any calculations or papers that show the effects of noise and/or pixel shape on the resolution.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Tuesday, April 22, 2003 2:27 PM To: Mike Bode; Microscopy-at-MSA.Microscopy.Com
I suppose that arguments can be made that Nyquist is optimistic, but it provides a good estimate of what is needed. Noise may well require higher sampling frequencies, but on a normal image (not darkfield or fluorescence or similar), the noise should be of the order of a few percent. I can't say if that has a significant influence on the sampling frequency.
I do agree with Dr. Marks on the image resolution improvement, if you have additional information, of course. We do precisely that with some of our filters. In general, however, if you do NOT have additional information, image processing is usually the selective destruction of unnecessary or unwanted information (my words :).
CDs are to my knowledge recorded at 44 kHZz, which is 2x 22 kHz, which again is not too far from twice the highest frequency one can hear. If I remember correctly, the upper limit is about 20 kHz for someone with good hearing.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Monday, April 21, 2003 11:49 AM To: L. D. Marks; Mike Bode Cc: Microscopy-at-MSA.Microscopy.Com
I have not followed the details of this thread, but there are a couple of important points that need clarification/correction otherwise some misconceptions may take root.
1) For an "ideal" camera, it is true that all one wants is the Nyquist sampling. However, a ficticious ideal camera has no noise, and also only measures the intensity at the very center of a given pixel. In reality you have a finite pixel so the image is an integral of the intensity across the pixel which introduces a sinc function envelope to the resolution (in reciprocal or Fourier space). In addition, if you have a finite readout noise (or statistical, i.e. measurement since there may not be that many photons/electrons) this is per pixel. Combining these two in general it is better to have something like 2-3 times oversampling (relative to the Nyquist sampling); the exact value depends upon the camera and the experiment.
2) It is correct that most of the simpler image manipulation procedures (e.g. deconvolution, sharpening) do not improve the signal-to-noise - no free lunch. However, there are some well known cases where resolution extension (e.g. the Gerschberg alogorithm) is known to work. These exploit additional information, for instance knowledge that certain regions have no intensity or (in the case of maximum-entropy based algorithms) the fact that the noise is statistical in character. This is not "something for nothing" - it is rather exploitation of additional information not apparent in the image but present in the physics of the imaging process to reduce the apparent noise. (Some of the more advanced books in the image processing literature or astronomy deal with this, you won't find too much information in the simpler books.)
----------------------------------------------- Laurence Marks Department of Materials Science and Engineering MSE Rm 2036 Cook Hall 2225 N Campus Drive Northwestern University Evanston, IL 60201, USA Tel: (847) 491-3996 Fax: (847) 491-7820 mailto:ldm-at-risc4.numis.nwu.edu http://www.numis.nwu.edu -----------------------------------------------
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Hello all,
I´m interested in labelling bacterial surfaces with colloidal gold for negative stain and TEM. So far negative stain works OK, also indirect immunofluorescence with a primary mouse monoclonal. However, secondary antibodies conjugated with 10-15 nm gold doesn´t work at all. We have adhered fimbriated E Coli to Formvar-coated gold grids with or without fixation in various concentrations of formaldehyde, labelled them and fixed with glutaraldehyde. Any tips and tricks?? Regards, Margaretha
--
Margaretha Lindroth, Ph D Phone: 4613-222616 Dept of Medical Microbiology Fax: 4613-224789 Faculty of Health Sciences e-mail: Margaretha.Lindroth-at-hul.liu.se Linköpings Universitet SE-581 85 Linköping
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Dear Ian MacLaren,
I would greatly appreciate your reponse. I believe you spent much time answering my stupid question.
In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this sphere is not a plane. Sometimes it brings some troubles to us. Let me show an example. If the specimen normal is NOT parallel to incident beam, or if the orientation is NOT exact, the intersections between relrod and sphere are not symmetric. In SAD pattern, not only intensity but also the distance is different, in other words, SAD pattern is NOT symmetric. On the other hand, FFT always symetric even though zone axis is not exact. This let me believe FFT and SAD is not so identical.
Thanks for your kind calculation. Near [111]Si orientation, we can find SAD pattern. As you said, it is not an exact zone axis. I took three patterns with different tilting angles (x=-9, +2, +8) and spent two weeks understanding these patterns. After calibrating the tilting angles by Kikuchi pattens, my indexing results let me believe they come from an identical reciprocal plane (130)*. I believe your calculation is right. I guess the reasons why it is [130] rather than [230] maybe come from two points. 1. beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. (640) spot is too far from transmitted spot. 2. The epitaxial phase is under distorted state. The real angle maybe different from those calculated based on bulk data.
Till now I still think FFT is not completely identical to SAD. I believe the discussion itself is significant. In my opinion, we can view Ewald sphere as a plane and based on this approximation we can think FFT=SAD. But for epitaxy, we can not ignore this approximation if we want to discuss some slight mismatch information. Maybe I made some mistakes in my analysis. I hope to hear different sounds and look forward to hearing from you again.
Thanks again,
Best wishes,
Han
} Dear Ming, } This problem is in principle quite simple. Any diffraction pattern can be } seen as a Fourier transform of the diffracting object. You can find this } in any undergraduate optics textbook. Normally in TEM you will then have } to convulute the periodicity of the periodicity of the object with a top- } hat function describing its very limited thickness. This leads to the } well-known reciprocal lattice rods for thin samples. } } So, an SAD pattern is a Fourier transform including specimen periodicity } and specimen thickness. } } An HRTEM image can also be Fourier transformed. As you should know, } however, the HRTEM image is strongly influenced by the contrast transfer } function (i.e. highly defocus dependent). That means that the intensities } in a Fourier-transform of a HRTEM image are strongly affected by image } defocus. You see this effect particularly strongly for amorphous material. } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } geometry, but may have rather different intensities in the different spots } since the latter is so focus-dependent. } } That said, in your case the main point is that the geometry of the pattern } is identical for SAD or for Fourier-transformed HRTEM. } } Now, if your orientation relationship is correct, then when you orient } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } matching the (111) Si planes. I reckon on a quick calculation that the } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much too } large to see the [110]beta diffraction pattern. But the axis [230]beta } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } diffraction pattern? At such a small angle off axis, you will still see } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } symmetric). } } Best wishes } } Ian MacLaren }
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The Pt-C replicas can be detached from the glass substrate by submerging the specimen at a shallow angle (} 45º) into a small petri dish containing 6 to 10% HF. If the concentration is too high, the replica may disintegrate into small pieces. To clean the replicas of any biological material adhering to the replica undersurface, you may have to transfer to 70% sulphuric acid. Another choice would be 1- 2 M NaOH. After a few hours, the replicas should be transferred to distilled water and then to 400 mesh TEM grids. Any remaining water should be removed with filter paper. A good technical reference is Zeile, U. (2000) Fundamentals of cryo preparation and replica technique. 7th Asia-Pacific Electron Microscopy Conference. Reprints are available from Bal-Tac. However, other methods to avoid the creation of artefacts in biological specimens is to use an Environmetal SEM (LV-SEM, VP-SEM) or the cryo method for rapid freezing (plunge freezing, jet-freezing, or high pressure freezing) and the use a freeze-fracture system for Pt-C replicas.
-- Kelly
} } Dear Steve } If it's Pt or Pt/C coating to remove replica from the glass you may } use some HF solution. I don't remember exact concentration, but I } would think, 10% (v/v) should work. If not, you may try higher } concentration. Please, keep in mind: HF is extremely aggressive } chemical: it's strong irritant and will corrode everything on it's way } including SS EM tweezers. It should be stored in the plastic (NO } GLASS) and used with precautions. Basically, you use the same } technique as for floating Formvar film except everything should be } plastic and HF solution instead of water. When replica is detached } from the glass, you may move it with Pt loop to the water to wash it a } few times and then mount on the grids. Good luck. Sergey. } } At 12:55 PM 4/21/2003, you wrote: } } } } } } } Dear all } } } } I have a user who wants to make TEM carbon platinum replicas of } } mammalian cells growing on glass slides with attached tissue tek } } culture chambers. He figures to fix the cells, dehydrate, critical } } point dry, then coat the slides (minus the growth chambers) and float } } the replicas off the slide the same way he floats formvar off a slide } } when making coated grids. In his first attempt, he is unable to } } float the metal coating off the slide. A query to this listserver } } quite a while back mentioned using hydrofluoric acid to remove the } } replica. does anyone have details of this procedure, or preferably } } an alternative procedure that will enable my user to make his replicas? } } } } Thanks for any insights in this regard }
Mike, You are correct in your statement about Nyquist & Shannon, but this is not in fact how a camera works. If you take an image I(r), and measure it ONLY at a set of points (and nowhere else) what you said is completely true. (This is the case that they considered.) However, what a camera actually does is measure the integrated intensity over a pixel. This integration is equivalent to a convolution in the image plane, i.e. it introduces an envelope that modifies what you have in reciprocal (Fourier) space. If it is a square pixel this is a sinc function along the x & y directions; if you had a round pixel it would be a Bessel-type function (J1(ar)/ar if I remember right). When you say 2% noise, you have also to be a bit careful. If the measurement noise for a single pixel is 2%, you will get 2% noise at the Nyquist limit. If you take a coarser frequency, say twice this (in the image) the average noise is 2%/sqrt(2) - less. (Similarly for other frequencies). This gives the classic 1/f noise.
On Tue, 22 Apr 2003, Mike Bode 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 } -----------------------------------------------------------------------. } } } Ok, John, maybe I AM not getting the point, but here is as I see it: } } 1) Both Nyquist and Shannon calculated the THEORETICAL sampling frequency to } measure a frequency without introducing artifacts. They both arrive at 2X. } In a practical setting (camera with noise), the frequency may well be } higher. How much a few percent noise will add to the sampling frequency I } don't know, but my guess is, it's not a lot. } } 2) You mentioned the new SACDs. They use a higher sampling frequency to be } able to reproduce higher acoustic frequencies because the claim is, that the } music sounds better then (which I can't make any statement about). However, } their own web site claims, that they can reproduce up to 100 kHz. With a 176 } kHz sampling frequency, they are staying BELOW Nyquist and claim they can } reproduce 100 kHz sound. I am not saying that they can or can't or that it } sounds better or worse, just an observation that the guys at Sony and } Philips use numbers similar to Nyquist. } } Maybe we should move this discussion off-line before we start boring people. } } I'd be interested to hear if you have any calculations or papers that show } the effects of noise and/or pixel shape on the resolution. } } mike } }
SUMMER I 2003 COURSE ANNOUNCEMENT - Scanning Electron Microscopy (BIO. 222-Section BA)
NASSAU COMMUNITY COLLEGE, Garden City, Long Island, New York
A five week, Summer Session I, 2003 semester, course in Biological Scanning Electron Microscopy is being offered by the Biology Department of Nassau Community College. This is a 4 credit course offered four days per week (Monday through Thursday) between the hours of 8:00 am and NOON. Classes will begin on May 27 and end on June 26, 2003.
This is a "hands-on" course emphasizing biological specimen preparation, student operation of the SEM (Hitachi S-2400) and the production of electron micrographs through the process of black & white photography and digital image capture, along with electron micrograph analysis. Students will work on a number of biological samples with the goal of producing a portfolio of high quality SEM photomicrographs.
The course is widely transferrable and the cost per credit is reasonable at $106 per credit (for Nassau County residents or New York State residents with a certificate of residency).
More information about the Bio-Imaging Center at NCC, course descriptions and syllabi, and student gallery of EM photomicrographs is available at our web site. The URL is {http://www.ncc.edu/users/becks} .
Interested individuals should register as soon as possible since the course is limited to a total enrollment of ten (10) students.
If you have further questions, you should e-mail me directly at the address below.
For information about mail or telephone registration (Dial-a-Course) point your browser to http://www.sunynassau.edu/courses/sum03/pdf/dialacourse.pdf . The phone registration option is available until 5/1/03 by calling 516-572-7131 or 7372 or 7425.
P.S. A Fall 2003 TEM course is also being offered (BIO 221 - Section E2) on Thursday evenings beginning at 5:30 PM.
Stephen J. Beck Professor Bio-Imaging Center/Electron Microscopy Department of Biology Nassau Community College Garden City, NY 11530 Voice Mail: (516) 572-7829 Email: {becks-at-sunynassau.edu} URL: {http://www.sunynassau.edu/webpages/biology/becks.htm}
I have included the formulas I used to calculate the sampling frequencies for digital microscopy on my webpage.
The calculation is based on the Rayleigh formula and the Nyquist sampling rate is twice the resolution given by the Rayleigh formula, the Rayleigh formula gives a better approximation for high N.A. lenses than the Abbe formula. For the wavelength I have used green light at a wavelength (lambda) of 0.520 micrometer (um). The image pixel-unit is calculated from the CCD pixel size and the total magnification.
Abbe formula for resolving power of a microscope: d = lambda / 2 * N.A. Rayleigh formula for resolving power of a microscope: d = 0.61 * lambda / N.A.
I use the Rayleigh formula in another form for my own clarity: Rayleigh (um) = 1.220 * lambda(um) / ( 2.0 * Numerical Aperture ) Nyquist samplingrate = 0.5 * Rayleigh / image pixel-unit
} From this formula you can understand that turning down the light of a microscope by lowering the voltage of the lightsource will reduce the resolving power. The light will become "reddish" and the wavelength will shift from 520 to +/- 800. Using more "blueish" ligh will improve the resolving power for the microscope. You should use neutral density filters to reduce the amount of light instead of lowerin the voltage of your lightsource.
Using cameras with smaller pixelsizes will allow a lower magnification, which will increase the amount of light falling on the CCD chip in fluorescence microscopy where the intensity (I) is given by:
I = N.A.^4 / Mag.^2
However larger CCD pixel sizes give you a bigger "bucket", which allows to capture more photons.
As usual there is a lot more to say, but I stop here.
P.S.: in the discussion of the appropriate sampling rate, people tend to forget that for sound one also has to sample the harmonics. Musical instruments do not create a "pure" sound, but create complex sounds, composed by interacting harmonics.
I think we definitely shouldn't move the discussion offline when it's getting interesting.
I don't understand Prof. Marks' point about noise and the need for oversampling.
The sampling theorem states that any scene with a maximum frequency can be represented without loss by samples taken at twice that frequency. (That means two samples per shortest wavelength in the original scene.) This requirement that the scene is bandlimited also applies to whatever noise is present in the original scene or is added somewhere in the recording process before the actual sampling takes place. The effect of the finite size of the detector pixels is simply to multiply the Fourier transform of the image with a sinc function (for square pixels). Since this sinc function doesn't go to zero below the Nyqist frequency no information is lost and the correct amplitudes of all Fourier components can be easily recovered. (I'm assuming that the physical pixel size is not bigger than the distance between samples, so no overlap.) Assuming this correction is done the sampling theorem seems to state that the original signal including whatever noise there is can be perfectly reconstructed from the mentioned samples. I don't see why noise would require oversampling as long as it is also bandlimited. Actually I don't see how you distinguish between noise and signal from just one image. The sampling theorem will reconstruct whatever it samples, no matter whether that happens to be signal or noise.
I've heard about the need to oversample in a different context, namely when images need to be interpoated in order to align and average them. I have equally big problems with that. I wonder if anybody can point me to a publication?
Philip
----- Original Message ----- } From: "L. D. Marks" {ldm-at-risc4.numis.nwu.edu} To: "Mike Bode" {mb-at-Soft-Imaging.com} Cc: "'Mardinly, John'" {john.mardinly-at-intel.com} ; "'Microscopy-at-MSA.Microscopy.Com'" } } } Mike, } You are correct in your statement about Nyquist & Shannon, but } this is not in fact how a camera works. If you take an image I(r), and } measure it ONLY at a set of points (and nowhere else) what you said } is completely true. (This is the case that they considered.) However, } what a camera actually does is measure the integrated intensity over a } pixel. This integration is equivalent to a convolution in the image plane, } i.e. it introduces an envelope that modifies what you have in reciprocal } (Fourier) space. If it is a square pixel this is a sinc function along the } x & y directions; if you had a round pixel it would be a Bessel-type } function (J1(ar)/ar if I remember right). } } } } Ok, John, maybe I AM not getting the point, but here is as I see it: } } } } 1) Both Nyquist and Shannon calculated the THEORETICAL sampling frequency to } } measure a frequency without introducing artifacts. They both arrive at 2X. } } In a practical setting (camera with noise), the frequency may well be } } higher. How much a few percent noise will add to the sampling frequency I } } don't know, but my guess is, it's not a lot.
as you are looking at the surface of the root, you can just fix the root in 95% Ethanol, then to several times 100% ethanol. Then either you can go CPD or HMDS before coating. Aldehyde fixation can preserve well the organells, but I dont think it matters so much for your goals. I know a SEM expert who used this method to study the morphology of tree buds.
Good luck
Haixin Xu (Ph.D) Biological Sciences University of Maryland Baltimore County On Tue, 22 Apr 2003, Tobias Baskin 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 } -----------------------------------------------------------------------. } } } Greetings, } Fixation will distort the organ, no matter what. But you } should still be able to compare your samples. Aldehydes give the } best fixation, as a rule, but there is no reason to use cacodylate } for your purpose (actually, some would say no reason ever, but that } is another story). Plant roots can be fixed in FAA, dehydrated to } absolute ethanol, critically point dried, coated with metal, and } viewed. As an alternative to FAA, you could use 4% paraformaldehyde } in say PBS (or the dilute buffer of your choice, near pH7), rinse in } the buffer, dehydrate etc as above. I expect that 2 h in 4% } paraformaldehyde fix would be fine. You can check for shrinkage and } other untoward affects by looking at your fixed or dehydrated roots } under darkfield. } } Hope this helps, } Tobias } } } } } } } } A friend from our Soil Science department wants to compare roots from live } } plants grown in good soil and dead plants grown in poisoned soil. So far } } we have been looking at them under dark-field optical microscopy. It } } would be good if we could look at them under SEM, but we don't have an } } ESEM, and we don't want the root shape to collapse under vacuum } } dessication. } } } } When I have searched for root fixation I come across things like } } glutaraldehyde and cacodylate buffer, and we'd like to avoid using these } } if possible. Does anyone know simpler ways of fixing the external shape } } of roots? } } } } It would be nice if we could do the same thing for moulds, to show } } visiting students "this is what you get if you leave bread lying around in } } your accommodation!". } } } } Any ideas, please? } } } } +-----------------------------------------+ } } 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 } } +----------------------------------------+ } } } -- } _ ____ __ ____ Tobias I. Baskin } / \ / / \ / \ \ 109 Tucker Hall } / / / / \ \ \ Biological Sciences } /_ / __ /__ \ \ \__ University of Missouri } / / / \ \ \ Columbia, MO USA } / / / \ \ \ 65211-7400 } / / ___ / \ \__/ \ ____ voice: 573-882-0173 } fax: 573-882-0123 } http://www.biosci.missouri.edu/Baskin/baskin_lab__home_page.htm }
If I only compare 2D primitive cell shapes between FFT and SAD, I think the statement I said last time is right, i.e. FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD.
Is it right? Thank you very much. I also look forward to hearing your kind comment.
Best wishes,
Han
} } } FFT=SAD is true if Ewald sphere is viewed as a plane. In this case, FFT ALWAYS same as } SAD. } } I'm afraid not. In fact, the FFT (of the image) is NEVER the same as the SAD. This is a } point that I have always tried to stress when teaching at the NCEM Summer Schools. } } The FFT (or image diffractogram) is always complex hermitian because it is the FT of a real } function (the image intensity) -- that means that the intensities of the FFT (or image } diffractogram) are always symmetric (as you point out) for ANY image at ANY specimen tilt or } ANY misorientation. } } For a crystalline specimen, producing strong Bragg spots in the diffraction pattern, the } spots in the FFT (or image diffractogram) are well-defined combinations of interferences of } the Bragg spots in the diffraction pattern. These interferences come from the } auto-correlation (a kind of convolution) of the (complex) Bragg spot amplitudes (after the } Bragg spot amplitudes have been modulated by the phase changes imposed by the objective } lens). For a brief discussion of how the spots in the FFT (or image diffractogram, or image } intensity spectrum) are formed from the Bragg spots in the diffraction pattern, see } "Resolution-damping functions in non-linear images", M.A. O'Keefe, in 37th Ann. Proc. EMSA, } San Antonio, Texas (1979) 556-557. } } Of course, none of the above helps you with your orientation problem -- sorry. } } Mike O'Keefe } } } $B4Z(J $BL-at-(J 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 Ian MacLaren, } } } } I would greatly appreciate your reponse. I believe you spent much time answering my } } stupid question. } } } } In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is true if Ewald } } sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this sphere is not } } a plane. Sometimes it brings some troubles to us. Let me show an example. If the specimen } } normal is NOT parallel to incident beam, or if the orientation is NOT exact, the } } intersections between relrod and sphere are not symmetric. In SAD pattern, not only } } intensity but also the distance is different, in other words, SAD pattern is NOT } } symmetric. On the other hand, FFT always symetric even though zone axis is not exact. } } This let me believe FFT and SAD is not so identical. } } } } Thanks for your kind calculation. Near [111]Si orientation, we can find SAD pattern. As } } you said, it is not an exact zone axis. I took three patterns with different tilting } } angles (x=-9, +2, +8) and spent two weeks understanding these patterns. After } } calibrating the tilting angles by Kikuchi pattens, my indexing results let me believe } } they come from an identical reciprocal plane (130)*. I believe your calculation is right. } } I guess the reasons why it is [130] rather than [230] maybe come from two points. 1. } } beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. (640) spot is } } too far from transmitted spot. 2. The epitaxial phase is under distorted state. The real } } angle maybe different from those calculated based on bulk data. } } } } Till now I still think FFT is not completely identical to SAD. I believe the discussion } } itself is significant. In my opinion, we can view Ewald sphere as a plane and based on } } this approximation we can think FFT=SAD. But for epitaxy, we can not ignore this } } approximation if we want to discuss some slight mismatch information. Maybe I made some } } mistakes in my analysis. I hope to hear different sounds and look forward to hearing } } from you again. } } } } Thanks again, } } } } Best wishes, } } } } Han } } } } } Dear Ming, } } } This problem is in principle quite simple. Any diffraction pattern can be } } } seen as a Fourier transform of the diffracting object. You can find this } } } in any undergraduate optics textbook. Normally in TEM you will then have } } } to convulute the periodicity of the periodicity of the object with a top- } } } hat function describing its very limited thickness. This leads to the } } } well-known reciprocal lattice rods for thin samples. } } } } } } So, an SAD pattern is a Fourier transform including specimen periodicity } } } and specimen thickness. } } } } } } An HRTEM image can also be Fourier transformed. As you should know, } } } however, the HRTEM image is strongly influenced by the contrast transfer } } } function (i.e. highly defocus dependent). That means that the intensities } } } in a Fourier-transform of a HRTEM image are strongly affected by image } } } defocus. You see this effect particularly strongly for amorphous material. } } } } } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } } } geometry, but may have rather different intensities in the different spots } } } since the latter is so focus-dependent. } } } } } } That said, in your case the main point is that the geometry of the pattern } } } is identical for SAD or for Fourier-transformed HRTEM. } } } } } } Now, if your orientation relationship is correct, then when you orient } } } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } } } matching the (111) Si planes. I reckon on a quick calculation that the } } } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much too } } } large to see the [110]beta diffraction pattern. But the axis [230]beta } } } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } } } diffraction pattern? At such a small angle off axis, you will still see } } } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } } } symmetric). } } } } } } Best wishes } } } } } } Ian MacLaren } } } }
*********************************** Dr. Ming HAN Nanomaterials Laboratory National Institute for Materials Science 3-13, Sakura Tsukuba 305-0003, Ibaraki Japan Tel: +81-29-863-5548 Fax: +81-29-863-5616 E-mail: HAN.Ming-at-nims.go.jp http://www.nims.go.jp/nano_char/index.html
-- I have been asked to under take a project looking at junctional complexes in the lens of mouse eye. I understand, from personal conversations, that lens is very difficult to fix and embed for TEM. A combination fixative, including Picric acid, is recommended in the literature (Sakai et al 1991). Do any of you listers have experience of handling and processing of mouse lens for ultrastructural TEM studies?
Thanks for your kind comments. It is the reason that I propose this question. Actually I can not tilt the specimen to get both exact zone axes for epitaxial phase and substrate.
Best wishes,
Han
} Han, } another point to consider is that in cases of mismatched epitaxy } such as this, there will inevitably be some tilt between the different } crystals, giving a small deviation from the nominal orientation relationship } (typically {5 degrees). This is a combination of misfit strain and the } dislocations which are present at interfacial steps, which produce a } low-angle grain boundary type misorientation. } } Richard } } _______________________________ } Richard Beanland } Analytical Services } Bookham Technology plc } Caswell, } Towcester, } Northamptonshire NN12 8EQ } UK } Tel: +44 (0) 1327 356362 } Fax: +44 (0) 1327 356775 } http://www.bookham.com } } } } -----Original Message----- } From: HAN.Ming-at-momokusa.nims.go.jp [mailto:HAN.Ming-at-momokusa.nims.go.jp] } Sent: 22 April 2003 16:51 } To: Microscopy-at-sparc5.microscopy.com } Subject: Re: EDP and FFT } } } ------------------------------------------------------------------------ } 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 Ian MacLaren, } } I would greatly appreciate your reponse. I believe you spent much time } answering my } stupid question. } } In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD is } true if Ewald } sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But this } sphere is not } a plane. Sometimes it brings some troubles to us. Let me show an example. If } the specimen } normal is NOT parallel to incident beam, or if the orientation is NOT exact, } the } intersections between relrod and sphere are not symmetric. In SAD pattern, } not only } intensity but also the distance is different, in other words, SAD pattern is } NOT } symmetric. On the other hand, FFT always symetric even though zone axis is } not exact. } This let me believe FFT and SAD is not so identical. } } Thanks for your kind calculation. Near [111]Si orientation, we can find SAD } pattern. As } you said, it is not an exact zone axis. I took three patterns with different } tilting } angles (x=-9, +2, +8) and spent two weeks understanding these patterns. } After } calibrating the tilting angles by Kikuchi pattens, my indexing results let } me believe } they come from an identical reciprocal plane (130)*. I believe your } calculation is right. } I guess the reasons why it is [130] rather than [230] maybe come from two } points. 1. } beta-FeSi2 is base-centered orthorhombic, (320) spot should be extincted. } (640) spot is } too far from transmitted spot. 2. The epitaxial phase is under distorted } state. The real } angle maybe different from those calculated based on bulk data. } } Till now I still think FFT is not completely identical to SAD. I believe the } discussion } itself is significant. In my opinion, we can view Ewald sphere as a plane } and based on } this approximation we can think FFT=SAD. But for epitaxy, we can not ignore } this } approximation if we want to discuss some slight mismatch information. Maybe } I made some } mistakes in my analysis. I hope to hear different sounds and look forward to } hearing } from you again. } } Thanks again, } } Best wishes, } } Han } } } } Dear Ming, } } This problem is in principle quite simple. Any diffraction pattern can be } } } seen as a Fourier transform of the diffracting object. You can find this } } in any undergraduate optics textbook. Normally in TEM you will then have } } to convulute the periodicity of the periodicity of the object with a top- } } hat function describing its very limited thickness. This leads to the } } well-known reciprocal lattice rods for thin samples. } } } } So, an SAD pattern is a Fourier transform including specimen periodicity } } and specimen thickness. } } } } An HRTEM image can also be Fourier transformed. As you should know, } } however, the HRTEM image is strongly influenced by the contrast transfer } } function (i.e. highly defocus dependent). That means that the intensities } } } in a Fourier-transform of a HRTEM image are strongly affected by image } } defocus. You see this effect particularly strongly for amorphous } material. } } } } So, an SAD pattern and a Fourier-transformed HRTEM image have the same } } geometry, but may have rather different intensities in the different spots } } } since the latter is so focus-dependent. } } } } That said, in your case the main point is that the geometry of the pattern } } } is identical for SAD or for Fourier-transformed HRTEM. } } } } Now, if your orientation relationship is correct, then when you orient } } along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are } } } matching the (111) Si planes. I reckon on a quick calculation that the } } angle between the (110)beta plane normal and [110]beta is 13.4 $BB0(J, much } too } } large to see the [110]beta diffraction pattern. But the axis [230]beta } } lies only 1.86$BB0(J from the (110)beta plane normal. Are you seeing this } } diffraction pattern? At such a small angle off axis, you will still see } } the diffraction pattern and an HRTEM image (even if it$BQT(J not very good or } } symmetric). } } } } Best wishes } } } } Ian MacLaren } } } } } } ======================================================================= } 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. }
Film Users: About a year ago we noticed a slight change in density of low dose micrographs on SO163 film which had been developed in full strength D19 for 12 minutes, the recommended protocol by Kodak. Specific information on the SO163 film change has been difficult to obtain but through a supplier we heard that there was a change in late 2001 (something to do with manufacturing process and raw material availability). It was suggested to us that a 10-15% change in developing time may be sufficient to compensate. To identify the film lots, if the first three digits of the eight digit emulsion number on the front of each multipak of film is 205 or higher, then that pack is "new" film. Additionally, we stumpled upon a couple of multipaks of SO163 film with an unusual emulsion number: the first four digits were 9801. This turned out to be a lot of the "old" or original film which was manufactured in Canada. Don
Donald Gantz Dept. Physiology & Biophysics Center Advanced Biomedical Research Boston Univ. School of Medicine Boston, MA 02118 Email: gantz-at-biophysics.bumc.bu.edu Phone: 617-638-4017 (voice mail)
The Facility for Electron Microscopy Research at McGill University is looking to purchase new web based calendar software for the reservation of instruments and equipment. Some of the needs that we have identified are:
- Pre-defined time-slots - Time slots that can be customized - Can create many reservation pages for all the instruments/equipment - Can customize time slots per instrument - Can assign individual user names and passwords for the booking of each instrument - Keep historical reservation information
Using the criteria listed above, I would appreciate any recommendations.
Thanks,
Kelly
--
S. Kelly Sears, Ph.D., B.F.A. Manager, Electron Microscopy Centre McGill University, 3640 University Street, Montreal, QC H3A 2B2 (T) 514.398.6334; (C) 514.576.1926; (F) 514.398.5047; (E) sksears-at-eps.mcgill.ca http://www.medicine.mcgill.ca/emcentre/s__kelly_sears.htm
Could someone please tell me what cellular components/substances that Toluidine Blue stains? (We use 1% Toluidine Blue O and 1% Sodium Tetraborate in 30% Ethanol on 1 micron-thick Durcupan sections.)
We are investigating histopathological changes in the mouse cochlea.
Thank you,
J.M. Lett
Harold W. Siebens Hearing Research Center Central Institute for the Deaf 4560 Clayton Avenue St. Louis, MO 63110
John, you are great to advertise himself. Is clear advertisement permitted on ListServer? Or opposite: responds on advertisement are not permitted? It sounds the case: I had troubles when respond on Cryo-EM course (Vancouver) advertisement. Have a good day. Sergey.
At 01:35 PM 4/22/03 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
I think, this discussion is very interesting and useful. Moreover: I think such discussions are most important things on our ListServer. Many thanks to all participants. Sergey.
At 03:00 PM 4/23/03 +0200, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Sergey Ryazantsev, Ph.D. Electron Microscopy Department of Biological Chemistry, School of Medicine University of California, Los Angeles Box 951737 Los Angeles, CA 90095-1737
120 film is a lot bigger than the little rectangles of x-ray film my dentist uses. Will the film even fit in the processor? I suggest you call Kodak on the phone and talk to them. If you want a machine to process roll film try a company called Jobo.
Geoff
Teresa Flores wrote:
} We have borrowed a Dent-X developer and have been trying to go online to } KODAK to see if previous users have imputed times for developing } Technical } Pan-120 Kodak film using a Dent-X automatic developer (Used in Dentist } office to developer x-rays). } There are three choices to choose from in the Dent-X auto-developer. } 4 & 1/2; 6 and Endo. } Any help would be appreciated, even from KODAK to let us know where we } could look up the correct time to use for the Technical Pan - 120 film } (used to take TEM photos in a Zeiss 109). } Any imput is greatly appreciated and will save rolls of film. } Teresa Flores } LSUHSC } EM Lab } Path Dept } New Orleans,LA } } } }
-- -- ********************************************** 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 **********************************************
We have borrowed a Dent-X developer and have been trying to go online to KODAK to see if previous users have imputed times for developing Technical Pan-120 Kodak film using a Dent-X automatic developer (Used in Dentist office to developer x-rays). There are three choices to choose from in the Dent-X auto-developer. 4 & 1/2; 6 and Endo. Any help would be appreciated, even from KODAK to let us know where we could look up the correct time to use for the Technical Pan - 120 film (used to take TEM photos in a Zeiss 109). Any imput is greatly appreciated and will save rolls of film. Teresa Flores LSUHSC EM Lab Path Dept New Orleans,LA
Having looked at several packages myself, I would appreciate any leads you are given that do not appear to the list. Here is a partial list of what I have examined so far that may match your criteria, though you did not specify what platform to run it on. In addition to your criteria we are looking for a package that does not require installation of a client, requires little administrative overhead does not cost a fortune, and requires only a few clicks to use. That said I have not found anything suitable, though the closest contender is perlcal. One other drawback to them all is in tracking sessions added in series. Suppose you sign-up for every M,W,F 3-4. This booking goes in as one "unit" If you later do not show up on day or need to adjust the time slot, all slots are affected. I have not found a way to individually edit a serial booking in any of the packages I have tried. Another nice feature is e-mail reminders. Anyway, there are a bunch out there. Good luck
Scott Whittaker Laboratories of Analytical Biology Smithsonian Institution National Museum of Natural History PO Box 37012 MRC104 Washington DC 20013-7012 202-357-1651
} } } "S. Kelly Sears" {sksears-at-eps.mcgill.ca} 04/23/03 03:50PM } } } ------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
The Facility for Electron Microscopy Research at McGill University is looking to purchase new web based calendar software for the reservation of instruments and equipment. Some of the needs that we have identified are:
- Pre-defined time-slots - Time slots that can be customized - Can create many reservation pages for all the instruments/equipment - Can customize time slots per instrument - Can assign individual user names and passwords for the booking of each instrument - Keep historical reservation information
Using the criteria listed above, I would appreciate any recommendations.
Thanks,
Kelly
--
S. Kelly Sears, Ph.D., B.F.A. Manager, Electron Microscopy Centre McGill University, 3640 University Street, Montreal, QC H3A 2B2 (T) 514.398.6334; (C) 514.576.1926; (F) 514.398.5047; (E) sksears-at-eps.mcgill.ca http://www.medicine.mcgill.ca/emcentre/s__kelly_sears.htm
Over the years I have had to gradually revised the policy on announcements of all courses.
Originally only "non-profit" courses were allowed. Non-profit meant (to me ) you may charge costs but don't make any money, but there were loop holes which were constantly cropping up. You can envision any number if you think about how to declare you don't make a profit on a short course.
To be honest it was beginning to be alot of hassel with individuals as well as organizations saying that they were just covering all costs. I'm neither an auditor nor a lawyer so , in the long run and to try to be fair to everyone I changed the rules to treat all short course/workshop announcements equally.
Thus, it is currently the policy to allow a "single" short posting of an announcement for a short courses of interest to the Listserver community. (See theListserver FAQ)
The benefits to the general community of allowing a single posting of a course/workshop in my opinion are sufficient to warrant this change.
I will use my judgement and if I perceive any individual or organization abusing this single short announcement policy then I will take appropriate steps.
} ------------------------------------------------------------------------ } 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 all, } Does anyone have a good contact for buying M-Bond 610 in Germany? } } Our previous contact seems to have gone dead. We ring the phone but } get no answer. This was Vishay measurements group. } } Any other ideas? } } Best wishes }
-- -- ********************************************** 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 **********************************************
We are considering the purchase of an Alto system and are interested in better understanding the capabilities and ease of use of these systems. Applications will involve, at least in part, polymers.
Feel free to contact me off-line.
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
------------------------------------------------------------------------ The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- [ From: Garber, Charles A. * EMC.Ver #3.1a ] --
Ian MacLaren wrote: ============================================================== Does anyone have a good contact for buying M-Bond 610 in Germany?
Our previous contact seems to have gone dead. We ring the phone but get no answer. This was Vishay measurements group.
Any other ideas? ============================================================== M-Bond 610 has been available in Germany for many years via or long time distributor:
Mr.Adi Hassel Ing.-Büro fur Prozeßtechnik und Instrumentelle Analytik Connollystraße 29 D-80809 München 40
I am sure he would be more than happy to help you with your requirement for M-Bond 610! Details on M-Bond 610 are given on URL http://www.2spi.com/catalog/spec_prep/glue.shtml
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 ######################## ============================================
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Au contraire my friend. We, at the Lab have Meeting Maker labwide. It is used for the reservation of meeting rooms and I think it would be great for the reservation of scope time. Didn't think of it until your posting. And there it was in the list of applications. I am checking into it today!
I have made a series of appointments, on my MM calendar, regarding a certain meeting I attend on Tuesday mornings. And I have gone in later to change some parameter. A box pops up asking me if this will affect this particular date or all future dates of this activity. Also, if the microscopist would like to have the investigator/scientist on hand while studying the sample, all one has to do is send a proposal to that person through Meeting Maker.
Annie --
+++++++++++++++++++++++++++++
R. Ann Bliss Technician, Chemistry and Materials Science Materials Science and Technology Division Lawrence Livermore National Laboratory
I have found a Roll-Film Apron (long plastic strip with wavy edges) for the 120 film that was made for the Kodak Roll-Film Tanks that are no longer available. It is in very good condition and will hold approx. 36 inches of film. If anyone, like Teresa Flores, would like to have it for the price of shipping or to TRADE for an Apron for 35mm film (mine are getting brittle) I will be willing to part with it.
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
I have found a Roll-Film Apron (long plastic strip with wavy edges) for the 120 film that was made for the Kodak Roll-Film Tanks that are no longer available. It is in very good condition and will hold approx. 36 inches of film. If anyone, like Teresa Flores, would like to have it for the price of shipping or to TRADE for an Apron for 35mm film (mine are getting brittle) I will be willing to part with it.
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
I have found a Roll-Film Apron (long plastic strip with wavy edges) for the 120 film that was made for the Kodak Roll-Film Tanks that are no longer available. It is in very good condition and will hold approx. 36 inches of film. If anyone, like Teresa Flores, would like to have it for the price of shipping or to TRADE for an Apron for 35mm film (mine are getting brittle) I will be willing to part with it.
Pat Connelly The University of Pennsylvania Department of Biology Philadelphia, PA 91904-6018 215-898-7145 psconnel-at-sas.upenn.edu
} On Tuesday, April 22, 2003, at 08:51 AM, Han wrote: } } In my opinion, FFT=SAD reqires an approximation. I agree that FFT=SAD } is true if Ewald } sphere is viewed as a plane. In this case, FFT ALWAYS same as SAD. But } this sphere is not } a plane. Sometimes it brings some troubles to us. Let me show an } example. If the specimen } normal is NOT parallel to incident beam, or if the orientation is NOT } exact, the } intersections between relrod and sphere are not symmetric. In SAD } pattern, not only } intensity but also the distance is different, in other words, SAD } pattern is NOT } symmetric. On the other hand, FFT always symetric even though zone } axis is not exact. } This let me believe FFT and SAD is not so identical. } Dear Han, In addition, there are effects due to dynamical and secondary scattering that change the diffraction amplitudes (both magnitude and phase) so that they are not identical to the FFT of the scattering potential (the diffracting object). Yours, Bill Tivol EM Scientist and Manager Cryo-Electron Microscopy Facility Broad Center, Mail Code 114-96 California Institute of Technology Pasadena CA 91125 (626) 395-8833 tivol-at-caltech.edu
I've been following the discussion of pixel number and empty mag with interest, although I haven't been able to pick out the take-home message (are 3 megapixels enough or do I need 5?).
Coincidently, during this discussion I was contacted by a large company that makes LCD computer monitors. At first they were merely interested in using one of my MicroAngela pictures to show off their new 9.2 megapixel LCD display. However, this evolved into a discussion of the usefulness of large, high-res displays for scientific use, such as microscopy.
I realized that I have a 1kx1k cooled CCD on my EFTEM, a couple of 3.2 megapixel cameras on light microscopes, my confocal users typically use 512x512 pixels, but we can up that, and I have completely forgotten the number of lines my A-to-D converter picks up from my FESEM, final size 3.2MB.
The first question is, are there cameras out there that people are using, such as on newer types of instruments, that approach anywhere near 9 megapixels?
Would there by any reason to have a 9.2 megapixel monitor (3840 x 2400 pixels), 20.2 inches, for viewing micrographs? Other than the fact that having a big, bright, expensive high-res monitor on your desk would make you smile, of course...!
Trying to apply some of Gary Radice's math, if a 3mp camera will give you a 7.8 inch print at 300 dpi, will that image displayed on your monitor at 72 dpi require at least a 32.5 inch monitor to see all the information?
Comments?
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* ****************************************************************************
Toluidine blue at highly alkaline pH is pretty non-specific. Most things are varing shades of blue. In dilute aqueous solutions tol blue stains a variety of intercellular secretory granules and extracellular components, depending on the (acidic) pH.
Geoff
Jaclynn Lett 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 **********************************************
We are considering the purchase of an Alto system and are interested in better understanding the capabilities and ease of use of these systems. Applications will involve, at least in part, polymers.
Feel free to contact me off-line.
Gary M. Brown ExxonMobil Chemical Company Baytown Polymers Center 5200 Bayway Drive Baytown, Texas 77520-2101 phone: (281) 834-2387 fax: (281) 834-2395 e-mail: Gary.M.Brown-at-ExxonMobil.com
I did as David suggested and discovered that the Hopkins calendar appears to be a development that is at least introduced here:
http://www.macromedia.com/devnet/mx/dreamweaver/articles/aspnet_calendar_02. html Hopkins might even do better than charge heavily for the code, but even if they did, it would be worth a try to see how their IT folks implemented such a nice method.
In response to Scott's attempts to find a program that permits action on elements of a series, I go back to Outlook and its ability to calendar via mail/Exchange with calendars in public folders in which resources may be scheduled. Wherever that is used, however, would be private rather than web-based.
Cheers,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: David Elliott Ph.D. [mailto:David.Elliott-at-yale.edu] Sent: Thursday, April 24, 2003 9:08 AM To: Microscopy-at-sparc5.microscopy.com
I would suggest going to http://www.hopkinsmedicine.org/micfac/reservations.cfm check out there page. It may work for you. David
} } } }
} } } } } } } } } } ------------------------------------------------------------------- -- -- } } } } } - } } } } } 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 Facility for Electron Microscopy Research at McGill University is } } } } } looking to purchase new web based calendar software for the } } } } } reservation of instruments and equipment. Some of the needs that we } } } } } have identified are: } } } } } } } } } } - Pre-defined time-slots } } } } } - Time slots that can be customized } } } } } - Can create many reservation pages for all the instruments/equipment } } } } } - Can customize time slots per instrument } } } } } - Can assign individual user names and passwords for the booking of } } } } } each instrument } } } } } - Keep historical reservation information } } } } } } } } } } Using the criteria listed above, I would appreciate any } } } } } recommendations. } } } } } } } } } } Thanks, } } } } } } } } } } Kelly } } } } } } } } } } } } } } } -- } } } } } S. Kelly Sears, Ph.D., B.F.A. } } } } } Manager, Electron Microscopy Centre } } } } } McGill University, 3640 University Street, Montreal, QC H3A 2B2 } } } } } (T) 514.398.6334; (C) 514.576.1926; (F) 514.398.5047; (E) } } } } } sksears-at-eps.mcgill.ca } } } } } http://www.medicine.mcgill.ca/emcentre/s__kelly_sears.htm
On Wednesday, April 23, 2003, at 04:08 PM, Robert Kayton wrote:
} A faculty member asked me if knew anything about an ultra-small gold } particle, for immunogold labeling in E.M. } } Any help is appreciated. } Dear Bob, EM supply vendors sell colloidal gold in various sizes, and this can be conjugated to antibodies for immunogold labeling. Depending on your requirements, you may be able to purchase secondary antibody already conjugated. (I assume that your antigen is not common enough that you could get primary antibody commercially.) 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
Another group on campus that is handling scheduling for a research boat and is using this software "csCalendar". http://www.cgiscript.net/cgi-script/csNews/csNews.cgi?database=cgi.db&command=viewone&id=35. I haven't tried this yet but we are going to. I'll let you know how it works. Price is sure right.
Owen
At 08:10 AM 4/24/2003 -0400, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Owen P. Mills Electron Optics Engineer Materials Science & Engineering Michigan Technological University Rm 512 M&M Bldg. Houghton, MI 49931 PH 906-369-1875 FAX 906-487-2934 mailto:opmills-at-mtu.edu http://www.mm.mtu.edu/~opmills
Oops - a 3840 x 2400 pixel, 20.2 inch display has more than 72 dpi. The 20.2 inch designation is diagonal, of course, and I didn't take the time to do the math, but the display has closer to 190 dpi.
Sorry!
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* ****************************************************************************
On Tuesday, April 22, 2003, at 03:39 PM, Hiromi Konishi wrote:
} Several weeks ago, I asked on references of dose measurement } in microscope. I am still looking for that. This time, I want to more } specify my question. I would appreciate any comments you have. } } Here, I simply consider current density instead of dose. } There is a relation between current densities on specimen (Is) } and Faraday cup (If). } } Is = If x M x M } } Where M is a magnification on the Faraday cup. } } The relation between magnifications on Faraday cup (M) and film } (m) are given by m/M = l/L where L indicates the distance between } Projection Lens and Faraday cup, and l indicates the distance } between Projection Lens and film. We know magnification on film } (m) and can measure the current density on Faraday cup, so we } can calculate the current density on the specimen. } } I am looking for a textbook or paper that described the method I } wrote above. } Please advise. } Dear Hiromi, I performed such a measurement a few years ago, but did not publish the results. Our shop fabricated a high-precision Faraday cup that could be mounted in the same plane as the film, so the mag calibrations taken on film would also apply to the FC. Since M is squared, it is important to use the calibrated mag, not just the nominal mag. It is also important to have a FC that has the correct aspect ratio so that backscattered electrons do not escape--a paper by J.N. Turner (and perhaps other authors) discusses this. I had a piece of low-Z material inserted into the bottom of the FC to reduce backscattering further; either carbon or polyethylene will do. I was able accurately to measure the area of the hole through which the electrons could enter the FC, which is, of course, important. 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
Using Tech Pan film there are a very wide range of results you can get depending on the developer, agitation and time.
Your best bet is to call Kodak support explain to them what you are doing and how you are processing the film. They can get you started with the right developer, time etc. With Tech Pan these you can spend a lot of time trying to get what you want by experimentation. You need to be very consistent with all your development technique.
USA Support phone numbers: http://www.kodak.com/global/en/service/contactKodak/kodakPhoneNumbers.jhtml# 0012
International contact phone numbers: http://www.kodak.com/include/international.shtml ----- Original Message ----- } From: "Teresa Flores" {tflore-at-lsuhsc.edu} To: "Histotechnologist communicat" {histonet-at-pathology.swmed.edu} Cc: {ListServer-at-MSA.Microscopy.Com} Sent: Wednesday, April 23, 2003 8:47 AM
Add one more to your list to check out... www-sched at http://wilfred.berkeley.edu/~gordon/www-sched-download
You can vew a real sample of it at: http://wilfred.berkeley.edu/~gordon/www-sched/sched.cgi?fac=test2 I won't give out the passwords to this one yet, as we are doing some work on it this weekend.
The new version allows multiple schedules for different instruments/rooms and has a really nice web based interface for administration. You can manage all the schedules, including customizing their appearance, schedulable hours, how many people can sign up on one spot... etc.
Let me know what you think, as we are doing final customizations to it this weekend. The older www-sched is stand alone, and is meant to work with one schedule, unless you are adept at Perl. Gordon
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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, 24 Apr 2003, Scott Whittaker 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 } -----------------------------------------------------------------------. } } } Having looked at several packages myself, I would appreciate any leads you } are given that do not appear to the list. Here is a partial list of what I } have examined so far that may match your criteria, though you did not } specify what platform to run it on. In addition to your criteria we are } looking for a package that does not require installation of a client, } requires little administrative overhead does not cost a fortune, and } requires only a few clicks to use. That said I have not found anything } suitable, though the closest contender is perlcal. One other drawback to } them all is in tracking sessions added in series. Suppose you sign-up for } every M,W,F 3-4. This booking goes in as one "unit" If you later do not show } up on day or need to adjust the time slot, all slots are affected. I have } not found a way to individually edit a serial booking in any of the packages } I have tried. Another nice feature is e-mail reminders. Anyway, there are a } bunch out there. Good luck } } Ical-http://www.brownbearsw.com/ical/icalpage.html unix/pc } Webcal-http://bulldog.tzo.org/webcal/webcal.html unix } perlcal-http://www.perlcal.com/ unix/pc } Meeting maker- http://www.meetingmaker.com/home.cfm pc } webevent-http://www.webevent.com/demo/index.html } } Scott Whittaker } Laboratories of Analytical Biology } Smithsonian Institution } National Museum of Natural History } PO Box 37012 MRC104 } Washington DC 20013-7012 } 202-357-1651 } } } } } } "S. Kelly Sears" {sksears-at-eps.mcgill.ca} 04/23/03 03:50PM } } } } ------------------------------------------------------------------------ } 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 Facility for Electron Microscopy Research at McGill University is } looking to purchase new web based calendar software for the reservation } of instruments and equipment. Some of the needs that we have identified } are: } } - Pre-defined time-slots } - Time slots that can be customized } - Can create many reservation pages for all the instruments/equipment } - Can customize time slots per instrument } - Can assign individual user names and passwords for the booking of each } instrument } - Keep historical reservation information } } Using the criteria listed above, I would appreciate any recommendations. } } Thanks, } } Kelly } } } -- } } S. Kelly Sears, Ph.D., B.F.A. } Manager, Electron Microscopy Centre } McGill University, 3640 University Street, Montreal, QC H3A 2B2 } (T) 514.398.6334; (C) 514.576.1926; (F) 514.398.5047; (E) } sksears-at-eps.mcgill.ca } http://www.medicine.mcgill.ca/emcentre/s__kelly_sears.htm } } } } }
Ian; I would have though that the electrons forming SAD spots also would have gone through the objective lens, and have similar CTF convolution, but the part of the specimen sampled would be different between FTF and SAD.
John Mardinly Intel
-----Original Message----- } From: Ian MacLaren [mailto:maclaren-at-tu-darmstadt.de] Sent: Tuesday, April 22, 2003 2:07 AM To: HAN.Ming-at-momokusa.nims.go.jp Cc: Microscopy Listserver
Dear Ming, This problem is in principle quite simple. Any diffraction pattern can be seen as a Fourier transform of the diffracting object. You can find this in any undergraduate optics textbook. Normally in TEM you will then have to convulute the periodicity of the periodicity of the object with a top- hat function describing its very limited thickness. This leads to the well-known reciprocal lattice rods for thin samples.
So, an SAD pattern is a Fourier transform including specimen periodicity and specimen thickness.
An HRTEM image can also be Fourier transformed. As you should know, however, the HRTEM image is strongly influenced by the contrast transfer function (i.e. highly defocus dependent). That means that the intensities in a Fourier-transform of a HRTEM image are strongly affected by image defocus. You see this effect particularly strongly for amorphous material.
So, an SAD pattern and a Fourier-transformed HRTEM image have the same geometry, but may have rather different intensities in the different spots since the latter is so focus-dependent.
That said, in your case the main point is that the geometry of the pattern is identical for SAD or for Fourier-transformed HRTEM.
Now, if your orientation relationship is correct, then when you orient along [111]Si then the FeSi2 cannot be on axis if the (110)beta planes are matching the (111) Si planes. I reckon on a quick calculation that the angle between the (110)beta plane normal and [110]beta is 13.4 °, much too large to see the [110]beta diffraction pattern. But the axis [230]beta lies only 1.86° from the (110)beta plane normal. Are you seeing this diffraction pattern? At such a small angle off axis, you will still see the diffraction pattern and an HRTEM image (even if it’s not very good or symmetric).
Best wishes
Ian MacLaren
------- Forwarded message ------- } From: HAN.Ming-at-momokusa.nims.go.jp To: Microscopy-at-sparc5.microscopy.com
The gold label in question is likely the 1.4nm Nanogold particle, available from Nanoprobes, Inc. and probably from other distributors. Nanoprobes Web address is http://www.nanoprobes.com
-- _______________________________________________________________________ Michael Nesson, Ph.D. Department of Biochemistry & Biophysics 2011 Ag&LS, Oregon State University, Corvallis, OR 97331-7305 (541)737-5245 FAX:(541)737-0481 nessonm-at-onid.orst.edu
The smallest gold particles known to me are "undecaGold" 0.8 nm diameter and "nanoGold", 1.4 nm diameter. Both do have reactive group to bind with different compounds (antibodies etc). I do believe both compounds are still manufactured by NanoProbe: http://www.nanoprobes.com Sergey
At 02:17 PM 4/24/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Hello Nestor Does these changes mean that any institution including commercial may post a single advertisement on any course/workshop? It would be nice to establish some rules how to post such things. I would suggest to use some template, like: Course's title, date, place, short description and reference to particular WEB-site or contact person and positively NO self-advertising or words how successful that course was last year. "Short" should be short, right? Thanks. Sergey
At 05:47 AM 4/24/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Not to beat a dead horse (no, not the Amray SEM--it is quite alive), does anyone have any experience with EDS detectors on the 1810/1910 flat lens Amray systems? I am trying to add EDS to this 1910FE system but am frustrated by WD, take-off angle and physical position.
I need to be able to do spot analysis at about 6mm, ROI analysis at 6-10mm and mapping at between 6-15mm. Is this possible or just a dream?
The flat lens has distinct advantages over conical lenses. But it also has disadvantages. Such is life...and SEMs.
All inputs, public and private are appreciated.
gary g. Microtechnics, Inc. http://www.microtechnics.com Granite Bay, CA 95746 916.791.8191
Hello everyone It's not absolutely necessary to spent money on expensive huge LCDs. I do find that even on my 19" LCD with 1280x1024 the letters sometime too small to read comfortably. In most cases we need more space, not resolution (we are talking about monitors). If you are running Win2K you may use two monitors simultaneously. They act altogether as one big monitor. It mean that you could place your menu bars in one place (monitor) and use another for fine work. If it's a case, people usually use expensive, good monitor as a working place and another cheaper (and usually smaller) to store icons, menu-bars etc... Mouse and other controls act as it's single monitor. Such set-up is very popular between graphic designers. Sergey
At 12:46 PM 4/24/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Have you tried changing your font size to larger than it is?
gary g.
At 06:43 PM 4/24/2003, you wrote:
} Hello everyone } It's not absolutely necessary to spent money on expensive huge LCDs. I do } find that even on my 19" LCD with 1280x1024 the letters sometime too small } to read comfortably. In most cases we need more space, not resolution (we } are talking about monitors). If you are running Win2K you may use two } monitors simultaneously. They act altogether as one big monitor. It mean } that you could place your menu bars in one place (monitor) and use another } for fine work. If it's a case, people usually use expensive, good monitor } as a working place and another cheaper (and usually smaller) to store } icons, menu-bars etc... Mouse and other controls act as it's single } monitor. Such set-up is very popular between graphic designers. Sergey } } At 12:46 PM 4/24/2003, you wrote: } } ------------------------------------------------------------------------ } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
It's actually quite easy to make colloidal gold with diameters between 3.5 and 15 nm using a method such as Slot, Geuze 1985, Europ. J. Cell Biology 38, 87-93.
If you want even smaller labels you need clusters such as "nanogold", which is about 1.5 nm in diameter (Hainfeld, Furuya 1992, J. Histochemistry and Cytochemistry 40, 177-184)
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: "Bill Tivol" {tivol-at-caltech.edu} To: {microscopy-at-sparc5.microscopy.com} Sent: Thursday, April 24, 2003 11:17 PM
Yes, I could, but, for instance on some WEB-pages you could meet very small fonts and nothing to do with that. Personally, I am really happy with my monitor. My point was, that sometime we need more space, not necessary so many pixels when we are talking about monitors. Two monitors also is very economical solution. Sergey.
At 09:48 PM 4/24/2003, you wrote: } Have you tried changing your font size to } larger than it is? } } gary g. } } } } At 06:43 PM 4/24/2003, you wrote: } } } Hello everyone } } It's not absolutely necessary to spent money on expensive huge LCDs. I do } } find that even on my 19" LCD with 1280x1024 the letters sometime too } } small to read comfortably. In most cases we need more space, not } } resolution (we are talking about monitors). If you are running Win2K you } } may use two monitors simultaneously. They act altogether as one big } } monitor. It mean that you could place your menu bars in one place } } (monitor) and use another for fine work. If it's a case, people usually } } use expensive, good monitor as a working place and another cheaper (and } } usually smaller) to store icons, menu-bars etc... Mouse and other } } controls act as it's single monitor. Such set-up is very popular between } } graphic designers. Sergey } } } } At 12:46 PM 4/24/2003, you wrote: } } } ------------------------------------------------------------------------ } } } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
_____________________________________
Sergey Ryazantsev Ph. D. Electron Microscopy UCLA School of Medicine Department of Biological Chemistry Box 951737 Los Angeles, CA 90095-1737
Hello: Well, our SEM with LaB6 started to behave very strange. Turning on high voltage creates an emission current even with cold filament. About 12 kV the curren is small so we can eventually work. Higher high voltage, lets say about 20 kV, causes so high emission current (with no filament heating, Filamet=0) that there is emergency turn off. Any hint or suggestion would be greatly appreciated. Maybe I should add that the filament works "only" two months.
I have several large monitors on my desk. Not so that I can look at images on the whole screen, but so that when I am looking at a high res image, I can still have all of the editing tools on the screen. You need more (sometimes, much more) space on your monitor that is needed for the image. David
On Thursday, April 24, 2003, at 03:46 PM, Tina Carvalho wrote:
} Would there by any reason to have a 9.2 megapixel monitor (3840 x 2400 } pixels), 20.2 inches, for viewing micrographs? Other than the fact that } having a big, bright, expensive high-res monitor on your desk would } make } you smile, of course...!
Yes, you're right. The diffraction pattern is in fact distorted by the objective lens spherical aberration, i.e. the phase distortion implied by the CTF is also a geometrical distortion in the pattern (wave optics and ray optics agree on this point).
But the amount of distortion in the pattern is typically tiny. It gets bigger for very high-order reflections, but is essentially not measurable under normal circumstances. What is important for imaging is that when the beams interfere to form the image, this tiny directional shift translates into very signifacant phase shifts of the Fourier components of the image.
For the story as relates to SADP, the details are covered well in Hirsch, section 1.8 'Accuracy of selected area diffraction'.
Regards,
Wharton **************************************************************** Wharton Sinkler, PhD. UOP LLC 25 E. Algonquin Rd. Des Plaines, IL 60017-5017 tel. 847-391-3878 fax. 847-391-3719 mailto Wharton.Sinkler-at-uop.com
-----Original Message----- } From: Mardinly, John [mailto:john.mardinly-at-intel.com] Sent: Thursday, April 24, 2003 7:13 PM To: ian.maclaren-at-physics.org; HAN.Ming-at-momokusa.nims.go.jp Cc: Microscopy Listserver
Dear Dr. Kayton:
Ultrasmall colloidal gold particles was first introduce by Dr. Jan Leunissen (The founder of Aurion) as a marker for immunogold labeling. Antibodies conjugated to +/- one nanometer gold particles became commercially available in late 80's. Currently there are several companies carrying this type of products with different trade names. The following is a list. Even though "Ultra small gold" was first and still is used by Aurion as the trade name for their products, the term has become popular and often been used casually to refer to the whole category of small gold conjugates.
Ultra Small Gold: Aurion Immunogold Reagents and Accessories (distributed in the US by Electron Microscopy Sciences, www.emsdiasum.com) AuroProbe One: Amersham Biosciences, (www.amersham.com) One nanometer gold: British BioCell International (distributed in the US by Ted Pella, www.tedpella.com) Nanogold, Nanoprobes (www.nanoprobes.com)
Hong
==================== Hong Yi Emory Neurology Microscopy Core Emory University School of Medicine 6215 Woodruff Memorial Research Building 1639 Pierce Drive Atlanta, GA 30322
List, To add to the discussion, ultrasmall gold has advantages over larger gold conjugates. The size of US-gold is on the order of the size of the antibody allowing much better penetration, especially in tissue such as vibratome sections of brain. It can also result in increased labeling density. The US-gold is then silver enhanced to increase its visibility in the TEM. Tom
Thomas Moninger (thomas-moninger-at-uiowa.edu) University of Iowa Central Microscopy Research Facility (www.uiowa.edu/~cemrf) View expressed are my own.
In one sentence, ultrasmall gold probe has much higher sensitivity (meaning that you can dilute your primary Ab an order of magnitude further down), and yet you can grow it by silver enhancement as big as you wish, even to be visible under LM. The developing part is now much easier than it used to be.
I thought this was unfair that noone mentioned Aurion (http://www.aurion.nl/). They make a line of ultrasmall (0.8 nm) conjugates which may actually be a better fit for a histo-related application. I of course have no material interest in Aurion or its US distributor, but their Goat Anti-Rabbit F(ab')2-Ultrasmall has really done wonders for two of my applications. You are welcome to contact me with more specific questions, Bob.
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
} } From: darryl krueger {dkruege-at-clemson.edu} } Subject: TEM single crystal sample } } I have a friend that is doing Diffraction, who is looking for a source for } single crystals on TEM grids. If there is source someone knows of could } you please let us know. TIA } } Darryl
Tina; That monitor has a 205 dpi pixel density, so the pixels are .0049 inches apart. The resolution of the human eye is approximately 1/60 of a degree but that is only at the center of the field of view. The density of rods and cones on the retina is less off axis, At 10 inches viewing distance (nearsighted people can get closer and farsighted and presbyopic people would be farther away, without glasses, but that is another optical element...). This works out to approximately .0029 inches, so you should be able to easily see the pixels when you are ten inches away from the monitor. However, I have difficulty holding my eyes 10 inches away from a monitor while operating a microscope... Another plus would be the ability to display six 1 megapixel images without any overlap, if you need that.
John Mardinly Intel
-----Original Message----- } From: Tina Carvalho [mailto:tina-at-pbrc.hawaii.edu] Sent: Thursday, April 24, 2003 12:46 PM To: Microscopy Listserver
Hi, All-
I've been following the discussion of pixel number and empty mag with interest, although I haven't been able to pick out the take-home message (are 3 megapixels enough or do I need 5?).
Coincidently, during this discussion I was contacted by a large company that makes LCD computer monitors. At first they were merely interested in using one of my MicroAngela pictures to show off their new 9.2 megapixel LCD display. However, this evolved into a discussion of the usefulness of large, high-res displays for scientific use, such as microscopy.
I realized that I have a 1kx1k cooled CCD on my EFTEM, a couple of 3.2 megapixel cameras on light microscopes, my confocal users typically use 512x512 pixels, but we can up that, and I have completely forgotten the number of lines my A-to-D converter picks up from my FESEM, final size 3.2MB.
The first question is, are there cameras out there that people are using, such as on newer types of instruments, that approach anywhere near 9 megapixels?
Would there by any reason to have a 9.2 megapixel monitor (3840 x 2400 pixels), 20.2 inches, for viewing micrographs? Other than the fact that having a big, bright, expensive high-res monitor on your desk would make you smile, of course...!
Trying to apply some of Gary Radice's math, if a 3mp camera will give you a 7.8 inch print at 300 dpi, will that image displayed on your monitor at 72 dpi require at least a 32.5 inch monitor to see all the information?
Comments?
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* ****************************************************************************
Greetings, Does anyone have a protocol for immunolabeling intracellular constituents in small A. thaliana seedlings? I would like to label microtubules in a whole mount for confocal microscopy, but I anticipate that those nasty cell walls will pose a problem. Is there an effective way to permeablize the little weeds or is this a silly approach? Thanks in advance for any advice. Regards, Karl G.
On Thursday, April 24, 2003, at 12:46 PM, Tina Carvalho wrote:
Dear Tina,
} I've been following the discussion of pixel number and empty mag with } interest, although I haven't been able to pick out the take-home } message } (are 3 megapixels enough or do I need 5?). } } Coincidently, during this discussion I was contacted by a large company } that makes LCD computer monitors. At first they were merely interested } in } using one of my MicroAngela pictures to show off their new 9.2 } megapixel } LCD display. However, this evolved into a discussion of the usefulness } of } large, high-res displays for scientific use, such as microscopy. } } I realized that I have a 1kx1k cooled CCD on my EFTEM, a couple of 3.2 } megapixel cameras on light microscopes, my confocal users typically use } 512x512 pixels, but we can up that, and I have completely forgotten the } number of lines my A-to-D converter picks up from my FESEM, final size } 3.2MB. } } The first question is, are there cameras out there that people are } using, } such as on newer types of instruments, that approach anywhere near 9 } megapixels?
Yes. We are getting a 4k x 4k CCD. } } Would there by any reason to have a 9.2 megapixel monitor (3840 x 2400 } pixels), 20.2 inches, for viewing micrographs? Other than the fact that } having a big, bright, expensive high-res monitor on your desk would } make } you smile, of course...!
Yes, again--assuming that you wanted to look at several pics at the same time. } } Trying to apply some of Gary Radice's math, if a 3mp camera will give } you } a 7.8 inch print at 300 dpi, will that image displayed on your monitor } at } 72 dpi require at least a 32.5 inch monitor to see all the information?
Yes. However, if the 3mp was in the usual 3:2 ratio, the camera would be 1414 x 2121 pixels, so the image could be displayed at full resolution with a large border. Either the monitor is, indeed, as large as you calculate, or the display is at } 72 dpi. You might want to zoom in on a detail that wouldn't have apparent pixillation on the monitor, so the extra size could be warranted for that. 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 Thursday, April 24, 2003, at 05:13 PM, Mardinly, John wrote:
} I would have though that the electrons forming SAD spots also would } have gone through the objective lens, and have similar CTF } convolution, but the part of the specimen sampled would be different } between FTF and SAD. } Dear John, The electrons do go through the objective lens, and the lens is at precisely the same current for both SAD and SA imaging modes, but the projector lenses image the back focal plane in SAD, and there is no CTF convolution. This can easily be seen by taking an ED pattern of something like Au, where the intensities of the spots do not have too much structure--the unit cell is pretty simple--and taking the FFT of the corresponding image. Even if the objective lens is a little out of focus, you will not see the diffraction spots disappear at the zeros of the CTF, but you will see it in the FFT. 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 smallest gold particles known to me are "undecaGold" 0.8 nm } diameter and "nanoGold", 1.4 nm diameter. } There is the problem, however, of seeing the gold in your specimen, and undecagold is not visible (unless it can be decorated). Furthermore, if the specimen is stained, the gold must be distinguished from the other heavy metals, so it must be large enough to have an identifiably distinct shape. If it just looks like the clusters of stain, it won't be very useful. 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
Karl, Oh this is not silly, you can do it. Of course, chewing up the cell wall takes its toll, but you can get some nice images. Several groups have published on this. You can look at Sugimoto et al 2000 Plant Physiology 124: 1493-1506 for a protocol that has been used fairly widely.
A couple of points.
If you grow arabidopsis on agar plates, keep the plates vertical to get straight roots, on the surface of the plate, and when their time has come pour the fix directly on the plates (put them horizontal first!). This is the gentlest way, the plants die happy, and they stay submerged. No need to mess about with vacuums. When you are ready to add antibody, a nice idea is to put a droplet, 50 microL say, on a piece of parafilm covering the bottom of a Petri dish. Then you can put 5 or so root tips in the same drop and lay the rest of the root and the cotyledons out across the parafilm. The cotyledons are handy for picking up the seedlings and moving them from solution to solution (ie for rinsing) so don't cut them off until you are ready to mount.
One change that I recommend from the Sugimoto protocol is to replace the EGTA in the fix buffer with a little bit of CaCl2 (say 0.5 mM). EGTA is toxic and rips calcium out of the membranes and generally wreaks havoc on the cell. Not a good idea.
Hope this helps. Please feel free to contact me if you have further questions.
As ever, Tobias
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Hi all, The discussion on camera pixel numbers has been interesting but I haven't the expertise to contribute anything but have learned something. Thanks to all contributors.
However, I did make the jump to dual monitors as Sergey describes and find it very helpful in several ways: 1) When using Photoshop I keep the tools and maybe another app windows on one monitor, and the image on the Photoshop desktop on the primary monitor. This is great also when you are documenting in another app every step in manipulating the image; 2) when working on more that one document; and 3) when answering email like this one.
I did run into one issue with using dual monitors. Depending on which type of video board you get, you could end up with the desktop spread over two monitors making mousing across a wide field an annoyance but allowing you to open two images to a fairly large size (depending on screen width) with no overlap. Other boards make one monitor the primary and the other a secondary so that the desktop is on one monitor and the other just a blank work area where you can open a second app or a duplicate app. An example of this is Matrox G450 versus Matrox G550, the former puts the desktop across both screens, the latter puts the desktop on the primary screen (of your choice). I still perfer the CRT for my primary Photoshop screen as I think it still gives slightly better colors than an LCD but the gap is closing fast.
Damian Neuberger
} } Hello everyone } It's not absolutely necessary to spent money on expensive huge LCDs. I do } find that even on my 19" LCD with 1280x1024 the letters sometime too small } to read comfortably. In most cases we need more space, not resolution (we } are talking about monitors). If you are running Win2K you may use two } monitors simultaneously. They act altogether as one big monitor. It mean } that you could place your menu bars in one place (monitor) and use another } for fine work. If it's a case, people usually use expensive, good monitor } as a working place and another cheaper (and usually smaller) to store } icons, menu-bars etc... Mouse and other controls act as it's single } monitor. Such set-up is very popular between graphic designers. Sergey
I have a friend that is doing Difraction, who is looking for a source for single crystals on TEM grids. If there is source someone knows of could you please let us know. TIA
Darryl Krueger wrote: ===================================================== I have a friend that is doing Difraction, who is looking for a source for single crystals on TEM grids. If there is source someone knows of could you please let us know. TIA ===================================================== Actually there are number of such "samples" depending on the spacing you are wanting to calibrate and measure. These are all available from SPI Supplies as well as some of our major competitors:
The single crystal catalase specimen is probably the most commonly requested , see URL http://www.2spi.com/catalog/standards/othertem11.shtml
Another popular such test sample is the molybdenum trioxide specimen, see URL http://www.2spi.com/catalog/standards/othertem19.shtml
We also have the chloro-Cu-phthalocyanine test specimen favored by some, see URL http://www.2spi.com/catalog/standards/othertem9.shtml
And finally, there is the gold crystal specimen, see URL http://www.2spi.com/catalog/standards/othertem4.shtml
Other single crystal specimens can be found on URL http://www.2spi.com/catalog/standards/othertem.html
I am sure that there are others but these seem to be the most popular of the various possibilities, perhaps because they are low cost and are reasonably robust and seem to last the typical user quite some time.
Chuck
============================================
Charles A. Garber, Ph. D. Ph: 1-610-436-5400 President 1-800-2424-SPI SPI SUPPLIES FAX: 1-610-436-5755 PO BOX 656 e-mail:cgarber-at-2spi.com West Chester, PA 19381-0656 USA Cust.Service: spi2spi-at-2spi.com
Look for us! ######################## WWW: http://www.2spi.com ######################## ============================================
In general a digital microsocope is calibrated by using grids and measuring the distance between two points on the grid. The sampling error made when calibrating the microscope and the time spent, made us think of another way of calibrating a microscope by using the physical proersties of the CCD grid of the camera and the properties of the digitizer/framegrabber.
I want to know if this procedure is reliable and which problems may arise ? The result of the calculation will be the "size" of one pixel and as such can be used to calibrate the system. This example is based on a traditional PAL video camera (NTSC differs) and a framegrabber, modern digital cameras will need a different approach of course.
First we need to determine the chip area used on the CCD for acquireing, we have to take care only to use the active chip area in our calculations and not the entire chip. The placement of the CCD chip in the lightbeam of the microsocope will have an influence on the final magnification also, and there is no "standard" position for various cameras and microscopes. In addition when a camera dosn't have rectangular pixels, we will have to take this into account.
e.g. A (B/W) camera with the following specifications and square pixels:
A 1/2 inch CCD and a CCD diameter of 18 mm with a 3/4 aspect ratio, an array size 800 x 581 pixels (of which 756 active horizontaly) will result in:
The effective area is 756 x 581 pixels and not 800 x 581, thus the aspect ratio is not 4/3 ( 768/576 ), so in the end this will give us a (square) pixel size of :
Which instrument do you use? Some manufacturers have a "overnight" heating system which runs the filament at half power when you have switched OFF the high voltage. In this way they keep the filament out gassed when it is not in operation, which is a very good idea.
Could this system have failed to switch over to normal filament operation when you switched the high voltage on?
Please tell us more?
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: "Witold Zielinski" {WIZIEL-at-INMAT.PW.EDU.PL} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, April 25, 2003 11:28 AM
I am looking for a RMC MT 6000-XL specimen holder (the whole thing with arc segment mount etc., second hand or new). Anybody an idea where I could purchase one?
I guess I did not read you mail as well as I thought. I would suggest you have a microscope (Philips or FEI) that always has a degree of filament heating even if the "filament" supply is turned off!
With that in mind I think you need to clean the cathode. I believe you have a whisker shorting out the bias circuit. If you have a whisker of material between the filament and the cathode cap (Whenelt, or whatever you call it) the bias circuit is no longer in operation and the current is massive. Add to that the stand-by heating and you will easily obtain an image. Push the high voltage up too high and the safety circuit will trip in as you will pull too much current.
Sorry I did not spot this when I first read your mail.
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: "Witold Zielinski" {WIZIEL-at-INMAT.PW.EDU.PL} To: {microscopy-at-sparc5.microscopy.com} Sent: Friday, April 25, 2003 11:28 AM
} From: Coetzee, Mr S. H Physics Science } Sent: Monday, April 28, 2003 12:24 PM } To: Listserver Microscopy (E-mail) } Subject: Needs assessment - EM project for kids } } } Dear All } I would love to call this e-mail: Needs assessment - video or DVD on EM } and LM under subject but due to juncmail filters I had to change } the subject to } get it to the list. } } Life is always interesting and full of fun when you are doing EM. } We are doing a needs assessment on the following: } "Introduction video/DVD and LM for kids" } We want to target 13 - 18 year old kids and hopefully be able to reach 1st } and second year students as well. } The idea is to improve the number of students in Science with the focus on } EM. Input will be greatly appreciated. } We need this input for fund application. } } } Stephan H Coetzee } Department of Physics } EMU } Private Bag 0704 } Gaborone, } Botswana } } Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} } } Telephone: (+267) 355 2462 } Fax: (+267) 3185 097 } Telephone: (+267) 355 0000 Switchboard } }
When you make an image, the electrons that are scattered by the sample in different directions have to be brought back together by the lens. The phase relation between the electrons that leave the sample in different directions is important. If it is wrong, the image will be wrong. The contrast transfer function tells you how the electrons in different directions are shifted in phase. It controls the quality of the image.
In diffraction, whether selected area diffraction or convergent beam diffraction (which would be a better choice nearly always), the electrons that leave the sample in different directions are never brought back together. They go to different places in the diffraction pattern. Therefore the phase shift has no effect on the pattern. The contrast transfer function plays no role. This is why a diffraction pattern is still good if the microscope is misaligned, but an image is greatly degraded if the microscope is misaligned.
Alwyn Eades -- .......... Alwyn Eades Department of Materials Science and Engineering Lehigh University 5 East Packer Avenue Bethlehem Pennsylvania 18015-3195 Phone 610 758 4231 Fax 610 758 4244 jae5-at-lehigh.edu
This is a commercial reply relative to the request from: Gary S. Laevsky, Ph.D. Research Associate The Scripps Research Institute " I'm looking for the "best" source for an objective warmer."
Below is a clip and paste from our brochure on objective heating. But first a word of caution to all on this subject.
Bioptechs developed the first patented commercial objective heating system over ten years ago. Now there are several companies making similar devices. There are several very important factors that should not be overlooked when selecting an objective warming device.
a. When heat is applied to an objective it must be applied efficiently to the central core (where the lenses are). This means that all of the heat must go into the objective. There can be no excess radiation. Reason, it will require X amount of energy to warm the objective above ambient plus a continuous amount of additional energy to overcome the heat-sink effect of the nosepiece. Therefore, if the heat is not applied efficiently, the heat that is not transferred to the objective will radiate out and upward toward your specimen. This excess heat can "cook" your specimen.
b. Not all objectives have the same thermal profile. Therefore, the control system must be able to compensate for these variations.
c. Not all objectives can be thermally regulated. When lenses are designed there is little, probably no, thought given by the manufacturers to the prospect of temperature control for live-cell observation. I think this is an oversight on their part but that is another story for another time. Further detailed information is available on the Bioptechs web site. http:.//www.bioptechs.com.
Snip The Problem: When high numeric aperture objectives are used to observe temperature sensitive specimens, heat from the specimen is transferred through the optical coupling medium (oil, glycerine, or water) to the colder objective. Therefore when live-cell microscopy requires the use of high numeric aperture lenses, it is necessary to control the temperature of the objective. The thermal mass of a fluid coupled objective is overwhelming when compared to the thermal mass of the cells. Unfortunately, microscope manufacturers do not offer temperature controlled objectives, nor do they consider the need for thermal regulation in the design of their objectives.
The Solution: To eliminate this thermal gradient, Bioptechs offers a patented Objective Heater System, which includes a heater/sensor and an electronic controller. The heater/sensor is comprised of an adjustable thin film heating band which surrounds 3/4 of the diameter of the upper region of the central retracting tube of the objective. A surface probe thermal sensor positioned in the gap formed between the ends of the heating band measures the temperature of the objective. This heater/sensor assembly is supported on an adjustable metal mounting to fit objectives ranging from 16 to 34mm in diameter. The heater loop requires a minimum of 5mm longitudinal, physical contact with the cylindrical objective surface. In some cases there may be a decorative collar on the objective which must be removed in order to permit adequate surface contact.
The Controller is specifically designed to slowly heat the objective over a fifteen-minute warm-up period, then hold the objective at the setpoint value within 0.2°C. The Controller operates from ambient to 50 °C and has special safety circuitry which utilizes a 0.9°C error window to shut down the Controller and sound an alarm if, for any reason, the temperature of the objective deviates after it has reached setpoint. A user calibration test is also built in to the Controller. End snip
Dan
-- Dan Focht Bioptechs 3560 Beck Road Butler, PA 16002 dan-at-bioptechs.com www.bioptechs.com V 724-282-7145 F 724-282-0745
I am not sure what you are getting at. Calibrating a microscope means, that you try to get rid of the errors and non-linearities of the optical system (lenses, etc.) and other influences that affect the true magnification of the system, so that you can do true measurements on an image. I have the impression, your calculation starts from known numbers about the chip size and ends up with a number for the pixel size, which you can also get from the vendor. If that is your goal, then yes, you can do it that way. However, you have now "calibrated" the chip or camera, but not the microscope.
If you think, that the errors introduced by the measurement of a standard are larger than those introduced by the lenses and other elements, you only need to take the total magnification between the sample and the camera chip and divide the pixel size by this number, and you arrive at the pixel calibration for the image.
Calibrating a microscope takes about 5 minutes, and you really don't need to redo it unless you change anything on the microscope (objectives, camera position, etc.).
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Peter Van Osta [mailto:pvosta-at-unionbio-eu.com] Sent: Monday, April 28, 2003 1:48 AM To: MSA
Hi,
In general a digital microsocope is calibrated by using grids and measuring the distance between two points on the grid. The sampling error made when calibrating the microscope and the time spent, made us think of another way of calibrating a microscope by using the physical proersties of the CCD grid of the camera and the properties of the digitizer/framegrabber.
I want to know if this procedure is reliable and which problems may arise ? The result of the calculation will be the "size" of one pixel and as such can be used to calibrate the system. This example is based on a traditional PAL video camera (NTSC differs) and a framegrabber, modern digital cameras will need a different approach of course.
First we need to determine the chip area used on the CCD for acquireing, we have to take care only to use the active chip area in our calculations and not the entire chip. The placement of the CCD chip in the lightbeam of the microsocope will have an influence on the final magnification also, and there is no "standard" position for various cameras and microscopes. In addition when a camera dosn't have rectangular pixels, we will have to take this into account.
e.g. A (B/W) camera with the following specifications and square pixels:
A 1/2 inch CCD and a CCD diameter of 18 mm with a 3/4 aspect ratio, an array size 800 x 581 pixels (of which 756 active horizontaly) will result in:
The effective area is 756 x 581 pixels and not 800 x 581, thus the aspect ratio is not 4/3 ( 768/576 ), so in the end this will give us a (square) pixel size of :
Does anyone know which company sell (make) a motorized Z-drive for light microscope? I am only interested in the simple add-on device to collect z-stacks of light images, I don't need a motorized stage. The microscope used is Olympus BX51.
Any shared information will be highly appreciated.
Xinran Liu
Xinran Liu, M.D., Ph.D. Center for Basic Neuroscience UT Southwestern Medical Center at Dallas Phone: 214-648-1830 Fax: 214-648-1801 E-mail: xinran.liu-at-utsouthwestern.edu
Hi Stephen, TEM is hard work, but while SEM is even harder to understand, with ESEM you can make movies - the perfect solution for we biologists! Here's a recipe that I just worked out.
You need to find some dehydrated, over the counter, crosslinked polyacrylamide that is used in diapers and in some crazy toys, like alligators that grow in the bathtub. I got mine in a bag of crystals sold for the purpose of helping me keep our Christmas tree sanguine for as long as possible. These crystals imbibe water and swell to at least 50x their original size. Thus, if you find the same raw material as was in my little bag, you will have to do as I did and crush a couple crystals with mortar and pestle until you have one VERY tiny crystal that you can place (with #5 tweezers) at the bottom of the smallest depression in your cold stage insert set. Settle the insert in the cold stage and bring the temperature to 5oC and the pressure to 840 Pascal. You should home the stage (WITHOUT ROTATION, of course), and you should connect focus to z ASAP, and finally raise the stage to as close to 10 mm as possible. You should be able to image the crystal and it should be on the order of 100-200 um in size. Set up the movie to capture 1 image every 30 seconds and start recording. Then raise the pressure to 860, 880, 900, and finally 920 Pascal at which point you should begin to see puddles (dark blotches in the image) begin to form. NOTE: you will have to continuously tweak brightness and contrast to keep the image balanced properly. Once the water begins to condense on the stage, you should soon see changes in the crystal. The crystal will grow to its maximum size (1-2mm) and then you can start reducing pressure from 920 back to 840 by increments of 20. Again keep tweaking the brightness and contrast to keep the image right, until you are back to your crystal. NOTE: If the crystal is strange enough, it will probably resemble a cartoon character or two during its ascent or descent from totally bilious.
Finally, the routine is not perfect yet, but I am working on it before I bring elementary kids in on the joke. On the first one I did, the crystal went through a growing flower stage on the way up and a dying chicken stage on the way down.
My next trick will not be so silly, though I do plan to try rice and unhusked wheat. I plan to reproduce something I have not yet seen but was done somewhere in Mississippi with an old Electroscan. A movie of opening and closing stomata on a leaf.
Finally, during my very shallow learning curve, I managed to figure out a way to pull the ribbon cable out of the stage, thus disconnecting it from it's brain. It has been repaired, and has worked well since, as long as I am on my knees when I close the stage door and home the stage before doing that.
Hope this works for you, if you can try it, and I would like your results too.
I am copying this to the Q200-600 users list as well.
Regards,
Fred Monson
Frederick C. Monson, PhD Center for Advanced Scientific Imaging Mail Drop: Geology West Chester University West Chester, PA, 19383 http://darwin.wcupa.edu/casi/ Phone/FAX: 610-738-0437
-----Original Message----- } From: Coetzee, Mr S. H Physics Science [mailto:COETZEES-at-mopipi.ub.bw] Sent: Monday, April 28, 2003 8:24 AM To: Microscopy-at-sparc5.microscopy.com
} From: Coetzee, Mr S. H Physics Science } Sent: Monday, April 28, 2003 12:24 PM } To: Listserver Microscopy (E-mail) } Subject: Needs assessment - EM project for kids } } } Dear All } I would love to call this e-mail: Needs assessment - video or DVD on EM } and LM under subject but due to juncmail filters I had to change } the subject to } get it to the list. } } Life is always interesting and full of fun when you are doing EM. } We are doing a needs assessment on the following: } "Introduction video/DVD and LM for kids" } We want to target 13 - 18 year old kids and hopefully be able to reach 1st } and second year students as well. } The idea is to improve the number of students in Science with the focus on } EM. Input will be greatly appreciated. } We need this input for fund application. } } } Stephan H Coetzee } Department of Physics } EMU } Private Bag 0704 } Gaborone, } Botswana } } Coetzees-at-mopipi.ub.bw {mailto:Coetzees-at-mopipi.ub.bw} } } Telephone: (+267) 355 2462 } Fax: (+267) 3185 097 } Telephone: (+267) 355 0000 Switchboard } }
Bill Usually the "silver enhancer" used to enlarge particle size to made it visible in EM or even in LM. This technique is quite popular when you use pre-embedding "staining" with antibodies. In this case the probe size is critical, so people tends to use the smallest possible probe. Another good things about "undecaGold" and "NanoGold" (I am not familiar with other brands) that it has only one specific reactive group, maleimide for instance. So, you may attach these particles very precisely: such probe is more "direct". Sergey
At 04:46 PM 4/25/03 -0700, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Anyone know where I could get someting coated with a thin layer of silicon dioxide?
A researcher here wants to coat a gold microelectrode with a 100 nm layer of silicon dioxide to use in a quartz crystal microbalance. He asked me because I have done other coating jobs for him, but I can't do silicon dioxide.
If you can do it or know where to go to get it done, let me know and I will pass the info on.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
Damian If I understand correctly, you are using "dual" monitor video-cards. Nevertheless is it true or not, you may use two independent video-cards. Personally, I like Nvidia Gforce line (ATI is good choice as well). I am using one AGP-video for primary monitor, and another PCI-video card - for secondary. Personally, I like to have Win toolbar on the single monitor, so I am using "smaller" monitor as a primary. I was suspicious about LCD monitors for years, but things are changed now. Samsung monitors are very good (no interest in company) and deliver very good image quality. My favorite setup is 17" (1280x1024) + 15" in portrait mode (780x1024). It comes with "Colorific (?)" software for color calibration. It's first monitor in my life, which delivers exact matching with color printer. Another good thing about LCD - it's not flickering. My eyes tend to get tired after a few hours of working with high-quality CRT monitor, with LCD I could work comfortably much longer. Have a good day. Sergey
At 11:51 PM 4/25/03 -0500, you wrote: } Hi all, } The discussion on camera pixel numbers has been interesting but I haven't } the expertise to contribute anything but have learned something. Thanks to } all contributors. } } However, I did make the jump to dual monitors as Sergey describes and find } it very helpful in several ways: 1) When using Photoshop I keep the tools } and maybe another app windows on one monitor, and the image on the Photoshop } desktop on the primary monitor. This is great also when you are documenting } in another app every step in manipulating the image; 2) when working on more } that one document; and 3) when answering email like this one. } } I did run into one issue with using dual monitors. Depending on which type } of video board you get, you could end up with the desktop spread over two } monitors making mousing across a wide field an annoyance but allowing you to } open two images to a fairly large size (depending on screen width) with no } overlap. Other boards make one monitor the primary and the other a } secondary so that the desktop is on one monitor and the other just a blank } work area where you can open a second app or a duplicate app. An example of } this is Matrox G450 versus Matrox G550, the former puts the desktop across } both screens, the latter puts the desktop on the primary screen (of your } choice). I still perfer the CRT for my primary Photoshop screen as I think } it still gives slightly better colors than an LCD but the gap is closing } fast. } } Damian Neuberger } } } } } } Hello everyone } } It's not absolutely necessary to spent money on expensive huge LCDs. I do } } find that even on my 19" LCD with 1280x1024 the letters sometime too small } } to read comfortably. In most cases we need more space, not resolution (we } } are talking about monitors). If you are running Win2K you may use two } } monitors simultaneously. They act altogether as one big monitor. It mean } } that you could place your menu bars in one place (monitor) and use another } } for fine work. If it's a case, people usually use expensive, good monitor } } as a working place and another cheaper (and usually smaller) to store } } icons, menu-bars etc... Mouse and other controls act as it's single } } monitor. Such set-up is very popular between graphic designers. Sergey
} I am looking for a RMC MT 6000-XL specimen holder (the whole thing with } arc segment mount etc., second hand or new). Anybody an idea where I } could purchase one? } Stefan -
Contact Dave Roberts of RMC (now a division of Boeckeler Instruments) at dave-at-boeckeler.com.
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
I need to perform plunge freezing of solutions for CryoTEM. I was wondering if anyone had some advice on how to handle these explosive cryogens while using the Gatan cryotransfer system.
Any and all advice would be welcome as well as additional protocals for plunge freezing.
What kind of something? I might be able to get a TEOS wafer or two (SiO2 on Si).
gary g.
At 01:46 PM 4/28/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
What kind of something? I might be able to get a TEOS wafer or two (SiO2 on Si).
gary g.
At 01:46 PM 4/28/2003, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
In a message dated 04/28/2003 10:05:23 AM US Mountain Standard Time, xinran.liu-at-utsouthwestern.edu writes:
{ { Does anyone know which company sell (make) a motorized Z-drive for light microscope? I am only interested in the simple add-on device to collect z-stacks of light images, I don't need a motorized stage. The microscope used is Olympus BX51.
Any shared information will be highly appreciated.
Xinran Liu } }
Xinran,
I would recommend either Ludl Electronic Products at {www.ludl.com} or Prior Scientific at {www.prior.com} .
On the Prior website, look under "focus controls." The product you're looking for is the "Laser Autofocus" system.
Best regards,
Bob
Bob Chiovetti GTI Microsystems Leica Regional Dealer Desert Southwest Region / USA
***Disclaimer: GTI Microsystems represents Ludl and Prior, along with several other manufacturers of peripheral microscope control systems, but not in your region. This information is offered only for your information. Please contact your local representing agency for more details.***
We have decided to install AC/DC field cancellations to improve the performance of our FE-TEM and FE-SEM. We also plan to put acoustic tiles on walls in both rooms. I was wondering if anyone in the community could share some experience with me. Off-line vendor responses are welcome.
**************************************** Chaoying Ni 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 *****************************************
Are we talking about calibrating the ability of the camera to measure distances (spatial resolution)?
What about using a graticule? Just use an appropriate graticule once with each lens and as long as you don't change the lenses or other system components it will 'never' need to be done again.
Graticules are expensive so for one-off jobs perhaps you could borrow?
There was a company in UK called Graticules Limited who have now been taken over by Pyser-SGI Limited (see http://www.vertmedia.co.uk/pyser/contact.htm) or write to graticules-at-pyser-sgi.com.
I am not connected with the company in any way and if anyone from the company is reading this - do you think you could make your website a little more user-friendly?
Also try searching on www.google.com using graticule(s) as a search term.
I hope this helps. Med vänliga hälsningar/With best regards
Gareth
Take a look at http://www.vardforbundet.se/ifbls2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
I have never made one for a microscope but I have made them for many other devices using 4 solid state relays, a power supply, stepper motor and two timing belt pulleys and a timing belt. The pulleys are select to give the resolution needed using a 200 step per revolution stepper motor.
The stepper motor is driven by a simple program from the printer port driving the realys.
The only unknown is how you trigger your camera. It really needs to be triggered by the computer as well after a few seconds wait for vibration to die down.
If you only need one building one of off the rack parts is probably the least expensive choice. If you need several replacing the solid state relays with discreet components will save money. I expect 10 or 15 would be the break even point.
I can help you with the design and software or I expect I can get them built for you and furnish the software.
If you are having them built or building them yourself consider what other features your would like in the future. A few dollars spent on a more complex controller at the start will save scrapping what you have an starting over.
Good Luck
Gordon Couger Stillwater, OK www.couger.com/gcouger
} From: "RCHIOVETTI-at-aol.com"-at-sparc5.microscopy.com : : In a message dated 04/28/2003 10:05:23 AM US Mountain Standard Time, : xinran.liu-at-utsouthwestern.edu writes: : : { { Does anyone know which company sell (make) a motorized Z-drive for light : microscope? I am only interested in the simple add-on device to collect : z-stacks of light images, I don't need a motorized stage. The microscope : used is Olympus BX51. : : Any shared information will be highly appreciated. : : Xinran Liu : } } : : Xinran, : : I would recommend either Ludl Electronic Products at {www.ludl.com} or Prior : Scientific at {www.prior.com} . : : On the Prior website, look under "focus controls." The product you're : looking for is the "Laser Autofocus" system. : : Best regards, : : Bob : : Bob Chiovetti : GTI Microsystems : Leica Regional Dealer : Desert Southwest Region / USA : : ***Disclaimer: GTI Microsystems represents Ludl and Prior, along with several : other manufacturers of peripheral microscope control systems, but not in your : region. This information is offered only for your information. Please : contact your local representing agency for more details.*** :
we sell a motorized Z-drive which only controls the focus. We also have controllers which allow to connect the Z-drive and a motorized X/Y- stage. You could use the Z-drive now and add the X/Y-stage later. Our system is based on a Märzhäuser Z-drive.
Please contact me if you would like to receive a quotation.
Listers, I have a person wanting to look at rhihzobium involvement in root nodules. What is the best way to prepare this sample? I'm most concerned
about how best to expose the fungal involvement with the least amount of
artifact. Any suggestions or references will be appreciated. Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
Hello, this is my first post to the list. I need to find the specification of a Philips both the CM10 and CM100 water chillers, so I can be sure that the replacements are up to the specification the microscopes require. If anyone could help, I would be very grateful,
You can buy stage micrometers for about $150 at the usual microscopy supply places. If I am not mistaken, they are even traceable. For an additional price you can also get certified micrometers.
I didn't get that far looking for graticule. "Stage micrometer" seems to be a better phrase to look for.
mike
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Gareth Morgan [mailto:Gareth.Morgan-at-labmed.ki.se] Sent: Tuesday, April 29, 2003 12:09 AM To: Microscopy-at-sparc5.microscopy.com
Hi
Are we talking about calibrating the ability of the camera to measure distances (spatial resolution)?
What about using a graticule? Just use an appropriate graticule once with each lens and as long as you don't change the lenses or other system components it will 'never' need to be done again.
Graticules are expensive so for one-off jobs perhaps you could borrow?
There was a company in UK called Graticules Limited who have now been taken over by Pyser-SGI Limited (see http://www.vertmedia.co.uk/pyser/contact.htm) or write to graticules-at-pyser-sgi.com.
I am not connected with the company in any way and if anyone from the company is reading this - do you think you could make your website a little more user-friendly?
Also try searching on www.google.com using graticule(s) as a search term.
I hope this helps. Med vänliga hälsningar/With best regards
Gareth
Take a look at http://www.vardforbundet.se/ifbls2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
I got 11,300 hits for graticule and 18,700 for stage micrometer. Why do we call the same thing such very different names? Med vänliga hälsningar/With best regards
Gareth
Take a look at http://www.vardforbundet.se/ifbls2004
Gareth Morgan MPhil MSc FIBMS, Department of Laboratory Medicine (Labmed), Karolinska Institutet, Huddinge Universitetssjukhus, F46 SE 141 86 Stockholm Sweden
OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet för klinisk patologi och cytologi.
NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. Clinical Histo- and Cytopathology Laboratory.
} We are doing a needs assessment on the following: } "Introduction video/DVD and LM for kids" } We want to target 13 - 18 year old kids and hopefully be able to reach 1st } and second year students as well. } The idea is to improve the number of students in Science with the focus on } EM. Input will be greatly appreciated. } We need this input for fund application.
Mr. Coetzee -
I'd like to help, but it isn't clear what sort of input you need. If a reviewed list of what is currently available on video & CD-ROM (mostly at the middle & secondary level) will help, I can send the updated MICRO information directly to you as an attachment. It will be available at the MICRO website (URL below) as a searchable database within the month.
Caroline
Caroline Schooley Project MICRO Coordinator Microscopy Society of America Box 117, 45301 Caspar Point Road Caspar, CA 95420 Phone/FAX (707)964-9460 Project MICRO: http://www.msa.microscopy.com/ProjectMicro/PMHomePage.html Intertidal invertebrates: http://www.fortbragg.k12.ca.us/AG/marinelab.html
You didn't say if you wanted TEM or SEM, but I've prepared nodules for both, so here goes:
For TEM, I usually just cut the nodule into small pieces and do a standard TEM prep---primary fix in buffered gluteraldehyde or paraformaldehyde/glut mixture, buffer washes, osmium post-fix, water washes, uranyl acetate enbloc stain/fix, washes, dehydration, infiltration (recommend Spurr's or Spurr's/Epon for better infiltration), embedding, and ultramicrotomy. Vacuum and/or microwave-assisted fixation/infiltration often helps get the reagents in. Sometimes the best part of the nodule is in the center, if you're looking for bacteria (rhizobial strains), otherwise you may find a lot of empty cells.
For SEM, I have done standard fixation (again, vacuum and/or microwave-assist helps with infiltration of reagents), up to about 90% ethanol. At this point, you can drop the nodules into liquid nitrogen until they stop fizzing, then slice/fracture with a razor blade, complete the ethanol dehydration, critical point dry, mount and view. You should be able to see good internal cell structure in the nodule, with lots of rhizobial bacteria/bacteroids inside the cavaties.
About the fungal involvement, if you mean the rhizobia they are actually nitrogen-fixing species of bacteria, such as Rhizobia or Bradyrhizobia. Did you perhaps mean mycorrhizae (hope I spelled that right)? If the latter, I have no knowledge or experience at viewing them in nodules, or if they're even present in nodules.
Hope this helps. Let me know if you need more detail.
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: Bill Chissoe [mailto:wchiss-at-ou.edu] Sent: Tuesday, April 29, 2003 8:52 AM To: MSA listserver
Listers, I have a person wanting to look at rhihzobium involvement in root nodules. What is the best way to prepare this sample? I'm most concerned
about how best to expose the fungal involvement with the least amount of
artifact. Any suggestions or references will be appreciated. Bill
-- ============================================================= Bill Chissoe III Electron Microscopist University of Oklahoma 770 Van Vleet Oval Norman, Ok. 73019 E-mail: wchiss-at-ou.edu Ph. (405)325-4391 =============================================================
Thank you for your reply. I am aware that calibrating a microscope with a graticule is the classiacla way to do. We use the apporach I mentioned before for our automated microsopy based system. It has multiple cameras and of course different objectives and other lenses. We sue the calculation I mentioed before to avoid the calibration step when we exchange lenses.
An automated dual (or triple) camera setup with 6 objectives and 3 intermediate lenses plus an additional lens in the camera coupler of one of the cameras would take us some time to calibrate the entire system. By suing th approach I mentioned before, we back-calculate the size an iamg epixel has when going trhough the entire system; objective -} intermediate lens -} camera -} framegrabber -} memory.
The formula I mentioned, allows to take into account the effect of the camera-framegrabber combination on the "physical" dimensions of an image pixel.
Best regards,
Peter Van Osta
=========================================== Gareth Morgan 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 } } Are we talking about calibrating the ability of the camera to measure } distances (spatial resolution)? } } What about using a graticule? Just use an appropriate graticule once with } each lens and as long as you don't change the lenses or other system } components it will 'never' need to be done again. } } Graticules are expensive so for one-off jobs perhaps you could borrow? } } There was a company in UK called Graticules Limited who have now been taken } over by Pyser-SGI Limited } (see http://www.vertmedia.co.uk/pyser/contact.htm) or write to } graticules-at-pyser-sgi.com. } } I am not connected with the company in any way and if anyone from the } company is reading this - do you think you could make your website a little } more user-friendly? } } Also try searching on www.google.com using graticule(s) as a search term. } } I hope this helps. } Med vänliga hälsningar/With best regards } } Gareth } } Take a look at http://www.vardforbundet.se/ifbls2004 } } http://www.ki.se/biomedlab } e-mail Gareth.Morgan-at-labmed.ki.se } } Tel +46 8 5858 1038 } Fax +46 8 5858 7730 } } Gareth Morgan MPhil MSc FIBMS, } Department of Laboratory Medicine (Labmed), } Karolinska Institutet, } Huddinge Universitetssjukhus, F46 } SE 141 86 Stockholm } Sweden } } OBS! Besöksadress: F-Huset, Forskningsgatan 2 F52, Rum 2.10. Laboratoriet } för klinisk patologi och cytologi. } } NB! Visiting address: Building F, Research Corridor 2 F52, Room 2.10. } Clinical Histo- and Cytopathology Laboratory.
Response to the following comments from Dr Ian S. Harper. Mon, 28 Apr 2003
Reply:
I am happy to take this opportunity to clarify any misconceptions regarding the PeCon objective heating product and Bioptechs Products. Bioptechs began producing a patented commercial objective heater in 1992. Although it is a common sense solution to an existing problem, it has been a difficult concept to mainstream into live-cell time lapse applications. Back in 1992 every major microscope manufacturer was petitioned to either adopt our objective heater or at least make objectives in a manner that an optimal heat transfer surface would be exposed. None of the manufacturers were interested. They all said there was not enough of a market for temperature controlled objectives to be worthwhile.
The Bioptechs system is designed to meet the specific needs of high resolution live-cell time lapse imagers and takes into account both the safety of the objective and the cells under investigation. It is not simply a heating device that mounts on an objective. Now that time lapse imaging has become a more commonly used methodology, there are several companies producing devices that claim to have similar capabilities as the Bioptechs system.
Be it known that none of these companies are affiliated in any way with Bioptechs. We do not re-label, re-bag, or permit any other representation of our products. We have a distributor network but they are not permitted to re-label our products.
PeCon has a very similar product in appearance sold for the same application as ours but that is as far as it goes.
I again suggest referring to my Monday April, 28th posting relative to important factors with respect to selecting an objective warmer. Especially the part concerning excess radiated heat.
Further info on this subject is available on our web site http://www.bioptechs.com.
Dan Focht
Original message:
} For general information: Objective heaters (heating device plus } controller) are commercially available from: } 1. Bioptechs } (http://www.bioptechs.com/Instructions/Obj_Htr_i/obj_htr_i.html). } The manufacturers recommend the removal of the objective sleeve so allow } adequate contact between heating loop and objective barrel. A thermistor } against the objective barrel senses the temp and is part of feedback } control. Two heater loop sizes avail. } } 2. PeCon (http://www.pe-con.de/PeCon/PeCon-News.htm).. for specific } Zeiss and Leica lenses. Design look rather like the Bioptechs system - } does anyone know if this is a rebadged product ? } } 3. Our local uni instrument maker designed and built a system for us } (now available through a company Scientific concepts } (http://www.scientificconcepts.com.au/) - 'scuse the plug but people } might be interested in other live cell imaging accessories they have. (I } get no kickbacks, in case you're wondering, am merely a satisfied } owner). I suggest you make contact with S.C. via the website as the } heater does not yet feature on the website. The design is based on a } metal heating cylinder made to fit each lens (not an issue really, as, } for example, all our Olympus immersion lenses have more or less the same } diameter). Contact area between heater and barrel is large, and when } coupled with a heating chamber at same temperature, I have measured } {0.2ºC variation over the radius of a 25mm coverslip. (Compare this to a } 5ºC under similar conditions with non-heated immersion lens !!!).
} 4. 20/20 Technology } http://www.20-20tech.com/objective_heater.htm } To fit Leica, Nikon and Olympus lenses. } } 5. Zeiss } http://www.zeiss.de/C12567BE0045ACF1/allBySubject/2E19B0FD92393E6CC1256CB400326879 } } 6. Scientifica Life Sciences - a low cost thermal foil wrapping for } objective heating. } http://www.scientifica.uk.com/a.html } http://www.scientifica.uk.com/a_categorydetails.php?id=6
} Dr Ian S. Harper, Director } Monash Micro Imaging } The Microscopy & Imaging Research Facility } Building 13C, Monash University, Clayton, VIC 3800, Australia
-- Dan Focht Bioptechs 3560 Beck Road Butler, PA 16002 dan-at-bioptechs.com www.bioptechs.com V 724-282-7145 F 724-282-0745
University of Washington Employment Opportunities Research Coordinator (Imaging)
Department: Biology Date Available: May 1, 2003 Closing Date: Open until filled.
General Duties/Description: Under faculty overview, oversee the operation of a Bio-Rad MRC-600 laser scanning confocal microscope (LSCM), a Bio-Rad Radiance 2000 multi-photon LSCM microscope, a Philips CM-100 transmission electron microscope (TEM), a JEOL 840A scanning electron microscope (SEM), a sample preparation lab and photographic facilities. In addition to oversight, this position has a substantial "hands-on" component.
Training/consultation responsibilities: train facility users (faculty, postdocs, grad students and technicians) on equipment and techniques; instruct users in photographic and darkroom techniques; analyze and solve problems in methodology, software and hardware; consult with users on technical aspects of research projects related to microscopy; perform TEM, SEM, or confocal microscopy for potential users to make sure the techniques suit their needs before they go through training on the instruments (this may include sectioning samples or teaching sectioning); and collaborate with faculty and students to carry out long-term research projects. Maintenance responsibilities: determine service needs and schedule repairs; perform maintenance and necessary adjustments to equipment; recommend the upgrade of facilities by evaluating new equipment and software for purchase; and install new equipment and software (includes configuring the computer). Assist with computer programs designed for image analysis, graphics output and three-dimensional reconstruction: provide technical support in software use age; customize software via configuration or macros to fit users' needs; write and update support documentation; troubleshoot problems; and assist users with variable computer skills. General responsibilities: ensure imaging suite and darkrooms are safe and uncontaminated environments; properly dispose of waste chemicals for imaging and photographic suite; and order and oversee use of supplies, calculate recharges for instrument usage.
Requirements: A Bachelor's degree and a minimum of 2 years' experience with TEM (fixation, embedding, sectioning, staining of biological samples); prior experience with SEM, confocal or multiphoton microscopy would be useful but not essential (we will provide training for the successful applicant); computer literate on DOS and Macintosh (able to write macros, experience with Photoshop and NIH Image desirable). An equivalent combination of education and experience may substitute for stated requirements.
Salary: Salary and benefits are competitive. Salary is commensurate with qualifications and experience.
Send a resume and letter describing relevant experience and education to:
Sharon Larsson, Administrator, Biology, Box 351800, University of Washington Seattle, WA 98195.
slarsson-at-u.washington.edu 206 685-8241
The University of Washington is an equal opportunity, affirmative action employer.
Can someone point me in the direction of a public domain program which can take as input raw (peak minus background) wds element peak intensities of standards and samples and convert the sample intensities into wt % concentrations?
thanks
rtch
-- Ritchie Sims Ph D Phone : 64 9 3737599 ext 87713 Microanalyst Fax : 64 9 3737435 Department of Geology email : r.sims-at-auckland.ac.nz The University of Auckland Private Bag 92019 Auckland New Zealand
There have been several listers who seem to be calling graticules and stage micrometers the same thing. In my experience in UK light microscopy a stage micrometer is a specific accurate 'ruler' which may be mounted on a light microscope slide. It is an absolute size such as 1 or 10mm and is subdivided for accurate absolute measurements. Most graticules are eyepiece devices which may have up to 100 linear divisions or be divided up in some other way such as by angles. You can calibrate an eyepiece graticule using a stage micrometer for a particular objective lens configuration but the graticule is still an arbitrary measure.
I suppose you could argue that a stage micrometer was a specific absolute graticule but that does not make the terms interchangeable. If I am being pedantic, I apologise.
Thanks for your attention
Malcolm Haswell e.m. unit University of Sunderland UK
We observed similar behaviour in gold labeling E.coli outer membrane proteins quite some time back. We were not succesful in trying to label even unfixed intact cells, whereas labeling was not a problem at all on ultrathin cryosections of aldehyde fixed cells. A small percentage of the intact cells would label however, being about the same number as the number of mutant cells with shorter LPS molecules. That was the basis of our conclusion that long carbohydrate chains of LPS caused steric hindrance in intact cells, whereas an approach 'from the side' on sections was just working fine.
You mention using 10/15 nm particles, I am not sure whether ultra small particles would have the same limitations if LPS carbohydrates are causing this phenomenon.
I´m interested in labelling bacterial surfaces with colloidal gold for negative stain and TEM. So far negative stain works OK, also indirect immunofluorescence with a primary mouse monoclonal. However, secondary antibodies conjugated with 10-15 nm gold doesn´t work at all. We have adhered fimbriated E Coli to Formvar-coated gold grids with or without fixation in various concentrations of formaldehyde, labelled them and fixed with glutaraldehyde. Any tips and tricks??
I would suggest a detailed site survey of your field problems before going ahead and buying any field cancelling system. You should really find out if fields are adversely affecting your scope rooms (preferably without the scopes there, I know this may be difficult!) and if the fields are of a nature which a field cancelling systems can deal with. This is because our field cancelling system was actually unable to cancel our largest field problem as the frequency was too low to damp (we ended up digging up and re-installing old mains cables to solve the problem). It is also important to realise that a field cancellation system can typically only damp fields at one point (where the measurement probe is), which begs the question where do you put it? At the objective lens? Or close to the gun/HT? Anybody who has tested the position effect more closely, I would love to hear your input!
Acoustic tiles are however a simpler matter and I would definately recomend you install these.
Hope that helped!
Dr M Weyland, 1851 Research Fellow, Electron Microscopy group --------------------------------------------------------------------------- Department of Materials Science & Metallurgy University of Cambridge Pembroke St Cambridge CB2 3QZ
--On 28 April 2003 22:11 -0400 Chaoying Ni {cni-at-udel.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 } -----------------------------------------------------------------------. } } } Hello all, } } We have decided to install AC/DC field cancellations to improve the } performance of our FE-TEM and FE-SEM. We also plan to put acoustic tiles } on walls in both rooms. I was wondering if anyone in the community could } share some experience with me. Off-line vendor responses are welcome. } } **************************************** } Chaoying Ni } 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 } ***************************************** }
Hi: We are currently investigating various freeze sub and freeze plunge systems and would appreciate any pros or cons people have towards the different systems. Vendors feel free to email me with your products and please include the prices. Thanks 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
Does anyone have any information about whether osmium tetroxide can be used in xylene?
In our clinical pathology EM lab I am frequently called upon to *rescue* a piece of tissue from a paraffin block and work it up for EM. The standard protocol is to deparaffinate the tissue in xylene, then hydrate the tissue to osmicate it then dehydrate it again to go on to infiltrating with resin and finally embedding/polymerizing.
I have a faint recollection of a discussion years ago that osmium could be suspended in xylene saving the whole hydration/osmication/dehydration step. Thus one would deparaffinate the tissue in xylene then osmicate in xylene and go on to infiltration from there.
Any comments?
Karen Bovard Creighton University Medical Center Department of Pathology Electron Microscopy Laboratory Omaha, Nebraska
Jon; If you have a Gatan duo mill, you can put a piece of quartz where the sample normally goes, and the sputtered SiO2 will deposit on your target substrate held near it.
John Mardinly Intel
-----Original Message----- } From: jmkrupp-at-cats.ucsc.edu [mailto:jmkrupp-at-cats.ucsc.edu] Sent: Monday, April 28, 2003 1:46 PM To: Microscopy-at-sparc5.microscopy.com
Anyone know where I could get someting coated with a thin layer of silicon dioxide?
A researcher here wants to coat a gold microelectrode with a 100 nm layer of silicon dioxide to use in a quartz crystal microbalance. He asked me because I have done other coating jobs for him, but I can't do silicon dioxide.
If you can do it or know where to go to get it done, let me know and I will pass the info on.
Thanks
Jonathan Krupp Microscopy & Imaging Lab University of California Santa Cruz, CA 95064 (831) 459-2477 jmkrupp-at-cats.ucsc.edu
I don't know about xylene but I do know it works well in 100% ethanol in many different freeze-substitution protocols. So even if xylene won't work, you might not have to do a full dehydration. But the strength of the osmication reaction in ethanol at room temperature is much stronger than in water so I would try low osmium concentrations.
At 11:23 AM 4/30/2003 -0500, you wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
Thomas E. Phillips, PhD Associate Professor of Biological Sciences Director, Molecular Cytology Core 3 Tucker Hall University of Missouri Columbia, MO 65211-7400
Hello all, I am trying to gather information about saving digital images. Currently I import all images to Quartz PCI and save them as TIFF files on our network. I would like to continue to do this however, the IT department is threatening to cut me off if I continue to use so much space on the server. We do occasional image analysis, and since JPEG compression is lossy and introduces artifacts, I am leary of heading in that direction. I thought that if I could convince them that this is the best practice for saving images we could come to some sort of agreement. Any information of how you handle saving SEM images would be helpful in my quest for peace. Thank you
~Amanda Marchese
PQ Corporation Research & Development Center 280 Cedar Grove Road Conshohocken PA 19428 (610) 651-4668
On Monday, April 28, 2003, at 07:11 PM, Chaoying Ni wrote:
} We have decided to install AC/DC field cancellations to improve the } performance of our FE-TEM and FE-SEM. We also plan to put acoustic } tiles } on walls in both rooms. I was wondering if anyone in the community } could } share some experience with me. Off-line vendor responses are welcome.
Dear Chaoying, We are installing a FE-TEM, and we had some acoustics problems (our fields were very low). We eliminated the acoustical vibrations at the source, and we hung thick acoustical curtains, which solved all but one problem, which we are still working on. During the course of our investigations, we found that acoustic tiles are very good for reducing noise in the few-hundred Hertz range and above, but they were poor for reducing lower frequencies. Depending on the noise spectrum at your facility, tiles may either be useful or useless. Getting rid of, say, 60 Hz noise requires mass, so even anechoic tiles will be impractical--they must be at least 1/4 wavelength thick, which is impractical. 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
Offer to split the cost or buy your IT department a new hard drive for the storage of your images.
Don't sacrifice image information if you don't have to.
Mike
} ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- ============================================================ Michael Dunlap office (530) 752-0284 University of California lab (530) 752-5489 Chemical Engineering & Material Science Fax (530) 752-9554 110A EU-II mrdunlap-at-ucdavis.edu One Shields Ave. http://www.chms.ucdavis.edu/ Davis CA, 95616 ============================================================
You can dissolve the osmium in xylene. Remove the paraffin first with xylene, then mix the osmium tetroxide in xylene directly before use. The osmium reduces over time, so the mixture does not store well. We buy 0.1gram ampules of crystalline osmium tetroxide and use it at a concentration of 1% in xylene. Time in osmium is dependent on the size of the sample: sections on slides-15 minutes, 1mm3 tissue blocks-45 min to 1 hour. Rinse afterwards with several changes of xylene, then on to propylene oxide and resin. To dispose of the osmium mixture, we combine it with corn oil (1 part Os waste: 2 parts corn oil).
Vicky
Victoria Madden Microscopy Services Laboratory Pathology and Laboratory Medicine University of North Carolina at Chapel Hill vmadden-at-med.unc.edu
Possible solutions with increasing costs would be to 1. Buy a larger hard drive to save the images on your computer and share it to the network (I've seen 120 GB drives for $99/50,000+ images, assuming 2 MB images), 2. Burn images to CD's, $2/300 images, 3. Buy a network storage device, like a SNAP server made by Quantum, Iomega, Dell, and others, 200 GB systems are about $1500-2000/ 100,000 images. Or tell your management you could go back to Polaroids at $2 to $3 an image versus about $.002/image on your hard drive. The network storage device is nice because the images are easily accessed by others without affecting the computer where the images are collected.
At 1:16 PM -0400 4/30/03, Amanda Marchese(RD) wrote: } ------------------------------------------------------------------------ } The Microscopy ListServer -- Sponsor: The Microscopy Society of America
-- David R. Hull NASA Glenn Research Center at Lewis Field Advanced Metallics Branch Mail Stop 49-1 21000 Brookpark Road Cleveland, OH 44135
Yes. Osmium dissloves in xylene. I have used a similar procedure. Biggest problem is measuring a tiny amount of osmium, since such a small amount is needed. Our Mettler is not in a hood, only an old swinging arm balance. I keep one vial of OsO4 crystals for such use. On the whole, over the years, such "rescued" tissue does not give good results and in most cases the results are uninterpretable and non-diagnostic. It is much more effective to put a snip of tisue for any case which might possibly need EM in EM fixative for potential use. The cost of processing one case for EM with poor results far outweighs the cost of a batch (or 2) of EM fixative and vials.
Becky Garrison Pathology Supervisor
-----Original Message----- } From: "kbovard-at-creighton.edu"-at-sparc5.microscopy.com [mailto:"kbovard-at-creighton.edu"-at-sparc5.microscopy.com] Sent: Wednesday, April 30, 2003 12:23 PM To: Microscopy-at-sparc5.microscopy.com
Does anyone have any information about whether osmium tetroxide can be used in xylene?
In our clinical pathology EM lab I am frequently called upon to *rescue* a piece of tissue from a paraffin block and work it up for EM. The standard protocol is to deparaffinate the tissue in xylene, then hydrate the tissue to osmicate it then dehydrate it again to go on to infiltrating with resin and finally embedding/polymerizing.
I have a faint recollection of a discussion years ago that osmium could be suspended in xylene saving the whole hydration/osmication/dehydration step. Thus one would deparaffinate the tissue in xylene then osmicate in xylene and go on to infiltration from there.
Any comments?
Karen Bovard Creighton University Medical Center Department of Pathology Electron Microscopy Laboratory Omaha, Nebraska
I'd be glad to explain how I handle this problem. It may not be extendable to your situation. But you can pick and choose if you find something useful.
My premise is that the digital images are priceless. Trying to re-create them is a horrible prospect. So redundancy is paramount. I have perhaps 10,000 image files, ranging from LM, macro and SEM plus film images that have been scanned. Some images are 5MB while others can be up to 100MB. They are all over the place, size-wise.
Once the images are captured on MO disk (Fujitsu magneto optical 3.5"), the images are transferred to RAID IDE drives on my main computer. These images are then processed and finalized. The original images are then sent to a set of Dell 715N NAS servers (1TB total) for fast access archiving. Then, I do periodic backups of the servers to Ultrium 1 LTO tape (redundant). Each media holds between 100-200MB, depending on compression ability. This is all done over 100BaseT LAN using Novastore NovaBack network software. Once a full backup is done, all the others are incremental ones. These tapes are stored off-site in a bonded vault (very low cost).
Final output is also stored on the NAS servers and LTO tapes but also on redundant DVD-R media (I used to use CDs).
So, depending on how much data you have that makes your IT people complain, consider getting a DVD-R burner, or CD burner as a minimum. Then, if necessary, get one NAS SNAP server. Dell, Maxtor, and others make these. They are about $1200 used, and vary in price based on total capacity (size of the four internal IDE drives). These run under Win2K but you can also store Mac data to them using DAVE and standard TCP/IP protocol.
Another option is to get two external Firewire or USB hard drives. Plug them in and store the data to both drives. These drives are pretty low cost. Furthermore, if you can accommodate them, removeable IDE drives are very handy. You install a tray in your system box and put a drive in a pull out drawer. They show up in the OS as a regular hard drive. I have about eight drives ranging from 30GB to 200GB. Again, the price of the lost data is $????
Hope that this gives you some ideas.
gary g. Microtechnics, Inc.
At 10:16 AM 4/30/2003, you wrote:
} Hello all, } I am trying to gather information about saving digital images. } Currently I import all images to Quartz PCI and save them as TIFF files on } our network. I would like to continue to do this however, the IT department } is threatening to cut me off if I continue to use so much space on the } server. We do occasional image analysis, and since JPEG compression is lossy } and introduces artifacts, I am leary of heading in that direction. } I thought that if I could convince them that this is the best practice for } saving images we could come to some sort of agreement. Any information of } how you handle saving SEM images would be helpful in my quest for peace. } Thank you } } ~Amanda Marchese } } } PQ Corporation } Research & Development Center } 280 Cedar Grove Road } Conshohocken PA 19428 } (610) 651-4668
if you want to or have to save the images on a server and can't afford to tolerate some changes or artifacts in the images, you only have the option for a loss-less compression. There are several options to do so, the most commonly known is probably the ZIP utilities, but there are others, for example JPEG2000. The last one actually offers both lossy and loss-less compression. I don't know if Quartz PCI supports these compressions or file formats, we have implemented in our software, specifically for this reason, a JPEG2000 compression within the TIF format, so you get the best of both worlds.
If you can use a different storage location, you can probably relax this a lot. For example, you can buy so-called "Network attached storage devices" (basically a rudimentary PC with huge hard disks, often with redundant storage systems) and simply hook them up to the network. Perhaps you can convince your IT department to let you buy them one of those and keep it with the servers? Because you are not running any applications on these boxes (except the administration interface), there is usually very little administration necessary.
Michael Bode, Ph.D. Soft Imaging System Corp. 12596 West Bayaud Avenue Suite 300 Lakewood, CO 80228 =================================== phone: (888) FIND SIS (303) 234-9270 fax: (303) 234-9271 email: mailto:info-at-soft-imaging.com web: http://www.soft-imaging.com ===================================
-----Original Message----- } From: Amanda Marchese(RD) [mailto:Amanda.Marchese-at-PQCorp.com] Sent: Wednesday, April 30, 2003 11:16 AM To: 'Microscopy-at-MSA.Microscopy.Com'
Hello all, I am trying to gather information about saving digital images. Currently I import all images to Quartz PCI and save them as TIFF files on our network. I would like to continue to do this however, the IT department is threatening to cut me off if I continue to use so much space on the server. We do occasional image analysis, and since JPEG compression is lossy and introduces artifacts, I am leary of heading in that direction. I thought that if I could convince them that this is the best practice for saving images we could come to some sort of agreement. Any information of how you handle saving SEM images would be helpful in my quest for peace. Thank you
~Amanda Marchese
PQ Corporation Research & Development Center 280 Cedar Grove Road Conshohocken PA 19428 (610) 651-4668
This is sooo strange to hear. Disk drives are so cheap, it is like $80 for a 120 Gigabyte drive now. Just buy your IT department a new drive for your own data, and tell them you'll throw in lunch.
You can also do the storage internally in your own lab. Buy a server with a large hard disk and dump your data onto there. Servers more than adequate for this run as low as $400-$800. Add a DVD burner and you can back up all your images onto more permanent storage on a monthly basis.
I've implemented the above in our laboratory. If you need help with it, contact me offline.
\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\/\ 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 Wed, 30 Apr 2003, Amanda Marchese(RD) 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 all, } I am trying to gather information about saving digital images. } Currently I import all images to Quartz PCI and save them as TIFF files on } our network. I would like to continue to do this however, the IT department } is threatening to cut me off if I continue to use so much space on the } server. We do occasional image analysis, and since JPEG compression is lossy } and introduces artifacts, I am leary of heading in that direction. } I thought that if I could convince them that this is the best practice for } saving images we could come to some sort of agreement. Any information of } how you handle saving SEM images would be helpful in my quest for peace. } Thank you } } ~Amanda Marchese } } } PQ Corporation } Research & Development Center } 280 Cedar Grove Road } Conshohocken PA 19428 } (610) 651-4668 } }