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Dear colleagues - I would like to bring to your attention a *12th
International Workshop on Piezoresponse Force Microscopy and
Electrochemical Strain Microscopy, *to be held at The Center for
Nanophase Materials Sciences, Oak Ridge National Laboratory, on March
4-8 this year. This workshop is organized by S.V Kalinin, A.P. Baddorf,
and A. Kholkin, and will feature a set of tutorials on advanced PFM and
ESM operation, and extensive hands-on labs. The attendance will be
limited to ~24 people. Please see below the information on the scope and
content of the school.

Yours truly

Sergei V. Kalinin

Piezoresponse Force Microscopy (PFM) has by now become one of the
dominant techniques for exploring ferroelectric, multiferroic, polar
macromolecular and biological, and ionic materials on the nanoscale. The
last several years have seen multiple breakthroughs in practice and
theory of PFM achieved in multiple research groups worldwide. This
workshop aims to provide an in-depth description of recent advances in
Piezoresponse Force Microscopy and to offer laboratory demonstrations
designed for advanced PFM practitioners. The workshop will introduce
basic principles of PFM operation, relevant instrumental aspects, and
image interpretation. The theory of cantilever dynamics, PFM contact
mechanics, and resolution theory, as well as their implications for
qualitative and quantitative data interpretation in PFM, will be
presented and illustrated experimentally. Recent technical advances in
PFM, including vector PFM, high-frequency PFM, band-excitation and dual
frequency resonancy tracking (DFRT) imaging, switching spectroscopy PFM
and imaging and polarization switching in liquids and vacuum, will be
presented. For ferroelectric materials, applications of PFM for domain
imaging, nucleation center mapping, and probing polarization dynamics in
thin films and capacitor structures will be discussed and demonstrated.
Finally, electromechanical probing of biological, electroactive polymer,
and soft-condensed matter systems beyond classical ferroelectric
applications will be described.

The 4 day workshop designed for advanced PFM users will include
tutorials and lectures given by leading PFM experts and experimental
hands-on tutorials on PFM imaging and spectroscopy. Ultimately, the goal
of the workshop is to build a network of advanced PFM practitioners to
promote rapid dissemination of theoretical knowledge, experimental
protocols, and novel technique development in this rapidly growing area,
as well as to establish links to areas such as energy storage and
conversion and information technology, macromolecular science and
electrochemistry.

The workshop is supported by the Center for Nanophase Materials Sciences
at Oak Ridge National Laboratory and the European Initial Training
Network (ITN) �Nanomotion.� Contact Sergei V. Kalinin (sergei2-at-ornl.gov
{mailto:sergei2-at-ornl.gov} ) for details.

**

*Piezoresponse Force Microscopy: Theory, Techniques, and Applications*

The workshop will open with a series of the tutorial talks on principles
and applications of PFM in the mornings and series of hands-on
experiments (samples brought by attendees) in the afternoon. The
hands-on training will be performed by the CNMS staff members of extant
platforms for the groups of ~3 people.

*Tentative list of tutorials:*

1. (Kholkin) Basic principles and history of PFM
2. (Kalinin) Contact mechanics, resolution theory, and interpretation of
PFM signal
3. (Kalinin) Cantilever dynamics and frequency dependent measurements in PFM
4. (Kalinin) Polarization switching in PFM: voltage and time spectroscopies
5. (Jesse) Band excitation methods and BE PFM
6. (Belianinov) Multivariate statistical methods: working with
multidimensional data
7. (Kalinin) Advanced PFM modes: spectroscopy and multivariate analysis
8. (Rodriguez) PFM and KPFM in liquids
9. (Maksymovych) Combined PFM and cAFM: ferroelectric tunneling and
domain walls
10. (Balke) Electrochemical Strain microscopy of Li-ion materials
11. (Kalinin) ESM of oxygen conductors and electrochemical effects in PFM
12. (Shvartsman) PFM of relaxor ferroelectrics
13. (Rodriguez) PFM of ferroelectric capacitors
14. (Martin) Domain structures and polarization dynamics in PZT and BFO
15. (Rodriguez) PFM of macromolecular and biological systems
16. (Jesse) Local thermal characterization
17. (Tselev) Microwave imaging

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Might try the following.

http://reindeergraphics.com/

Chrs&HNY,

Fred Monson

Frederick C Monson, PhD
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pannsylvania
West Chester, PA, 1938
610-738-0437
fmonson-at-wcupa.edu
________________________________________
X-from: oshel1pe-at-cmich.edu [oshel1pe-at-cmich.edu]
Sent: Wednesday, January 02, 2013 11:12 AM
To: Monson, Frederick

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} realname - Ken Hayes
} Email - khayes-at-firstsolar.com
} EDUCATION - Graduate College
} SUBJECT_OF_QUESTION - HRTEM Image Processing
} QUESTION - I collect a large number of HRTEM images in my job. I
} currently use ImageJ to view and process them. I am looking for a
} user friendly package softare package for HRTEM image processing
} with an emphesis on FFT pattern solving. the OS is Windows 7. Can
} you direct me to a list of recommended softare?
} Thanks.

--

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Dear Colleagues,
We are running a four day training course in Atomic Force Microscopy over�Easter�this year.
This is the second time we've run the course, and the students were�.very positive about the first edition.
The course will be held in our laboratory in Porto, Portugal, with two instruments for the students to use. The course will include days of theory and two days of practice, covering both Image acquisition and data analysis. The course will take place between the 25th and the 28th of March 2013.2. Places are very �limited with just a few remaining, so interested students are encouraged to reserve a place as soon as possible, but definitely before 5th January.�
Visit�http://bit.ly/TuuO2 for more information. Enquiries and reservations can be made by emailing afmhelp-at-gmail.com

____________________________________________________________________________
Atomic Force Microscopy
Dr Peter Eaton and Dr Paul West
Available through all good bookshops, or direct from Oxford University Press at:
http://ukcatalogue.oup.com/product/780199570454..do http://afmhelp.com

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Dear Colleagues, Apologies for the previous email which said that application for our AFM training course must be made by January the 5th. There was were some formatting errors in the email. The correct deadline is *January 15th*.
Regards,
Pete.

____________________________________________________________________________
Atomic Force Microscopy
Dr Peter Eaton and Dr Paul West
Available through all good bookshops, or direct from Oxford University Press at:
http://ukcatalogue.oup.com/product/9780199570454.do http://afmhelp.com

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From noreply-at-speed-magazin.com Tue Jan 8 17:47:21 2013
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Hi Ben,

I've done this with a Canon camera. These are a bit easier because the
phototube can be purchased, and Canon has built in features for
tethering the camera to the computer, live focus, etc.

However, it looks like you're on the right track. The sensor has to be
160mm from the back of the objectives. Otherwise, the focus point will
change for the objective when a specimen is focused by eye. It's best to
devise a means to slide the camera up and down along 2 tubes (a wider
diameter tube and a narrower diameter tube), or to order a focusing tube
from Thorlabs. The quick and dirty way is to use one tube at a narrower
diameter and a larger diameter "sleeve." Have a machine shop drill
threaded holes into the sleeve for thumb screws (or, if you're like me,
you do it yourself with a drill press and hand threading device). It may
be a bit difficult to get two different sized tubes that are meant to
fit to each other, but you can try Thorlabs or Edmund Optics.

Mount your camera, focus by eye, and then, while the camera is giving
you a live view, adjust the outer sleeve up or down until the image is
in focus. Then tighten the thumb screws against the inner tube at that
position.

The image may not fill the sensor. Who cares. I suspect you will have
plenty of pixel coverage, even for Nyquist rates. Simply crop the images
after these are acquired, best done via an automated routine in
Photoshop or other imaging program. In that way, the crop will always
remain the same. Anyway, you'll likely get darkening at the edges
(vignetting), some barrel distortion, out-of-focus areas, etc., so even
if the entire sensor is filled, you'll still want to exclude image edges.

If it goes the other way and the sensor is overfilled, then you will
have to drop a lens into the tube.

If you could find out the focal length of the tube lens in that model of
Leica, then you could calculate coverage of the sensor (there's a lens
after the objective along the light path). Or, you can turn off all the
lights and hold a piece of paper where the sensor *should* be to get the
diameter of the virtual image when focused.

Good luck!

Jerry

On 1/3/2013 4:50 AM, ben.micklem-at-pharm.ox.ac.uk wrote:
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} Dear List,
}
} I am very interested in hearing from anyone who has mounted a Nikon 1
} series compact system camera on a light microscope.
}
} I am considering trying a Nikon V1 camera and a cheap LCD monitor via HDMI
} for a low-budget imaging system on an old Leitz 160mm tube length
} microscope. I have a trinocular head (38mm photoport), and an adapter for
} it to C-mount (with no optics in it). Is this adapter the 1x, which I
} understand is suitable for covering a 1" sensor?
}
} If so, all I need is a c-mount to 1 series mount adapter, and I should be
} ready to go?
}
} Best regards,
}
}
} Ben
}
} --
} Research Support Manager
} MRC Anatomical Neuropharmacology Unit, Mansfield Road,
} Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271649
} {http://www.mrc.ox.ac.uk/}
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--
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Scientific Imaging & Image Analysis Consultant
Technical Writer / Regulatory Consultant
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Author: �Scientific Imaging with Photoshop: Methods, Measurement and Output�

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***************************************************************************************
Forwarded from "Ask a Microscopist"
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****************************************************************************************
} realname - Ekaterina
} Email - Eyarunov-at-uwyo.edu
} EDUCATION - Graduate College
} QUESTION - Hello! I am having problems with sectioning my blocks for
} TEM. I have blocks of cells embedded into Lowicryl resin and I am
} trying to obtain sections 50-70 nm thick. However my sections are
} very wrinkled and I cannot use them for further immunoEM. What would
} be the reasons that can be fixed to avoid wrinkling? Temperature,
} speed of cutting, etc?
} Thank you for you help,
} Ekaterina

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Hello All,

Registration for The Histochemical Society course in
Immunohistochemistry and Microscopy at MBL in Woods Hole, MA has been
extended until JANUARY 25, 2013.

Application and complete information on website:
http://hermes.mbl.edu/education/courses/special_topics/ihcm.html

The course is four full days and evenings (11 hours daily) of lecture
and laboratory sessions with experts in the field of
immunohistochemistry (IHC) and microscopy.

FASEB/MARC AND HCS TRAVEL AWARDS for THE COURSE

The FASEB MARC Program provides funding for travel awards to encourage
and support the participation of underrepresented students and
postdoctoral fellows in the HCS IHCM course. The deadline for applying
is February 9, 2013:
http://www.faseb.org/MARC-and-Professional-Development/Travel-Awards.aspx

The Histochemical Society offers travel awards to attend the HCS Course.
The deadline for applying is January 31, 2013:
http://histochemicalsociety.org/Awards/Students---Post-Docs.aspx

Please email if there are any questions, happy to help.

Kind regards,
Meg McGough
mmcgough-at-histochemicalsociety.org

==============================Original Headers==============================
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Email: LES-at-ZSGENETICS.COM
Name: Larry Scipioni

Organization: ZS Genetics

Title-Subject: [Filtered] Averaging STEM-HAADF images

Message: Dear Steven,
If you are looking to align and average a stack of noisy images, StackReg works great in ImageJ.
Then just use the "Z Project" command to average it all.
Also, if you use "Save as" tif in DM on a stack, it will produce a multipage tif that opens up
directly in ImageJ as a stack, ready for averaging.

Steven Spurgeon wrote:

I have a series of STEM-HAADF images taken at 40 micro-sec time
intervals that I would like to cross-correlate and average. Is there
any freely available software to do this, such as a plugin for ImageJ
or DigitalMicrograph? Also, does anyone know of a script to batch
export a series of slices from an image in DM?

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Email: tkremer-at-ipstesting.com
Name: Tom Kremer

Organization: IPS Testing

Title-Subject: [Filtered] Leitz 1512 Microtome

Message: I inherited a Leitz 1512 rotary microtome. It's in excellent shape but without any knives.
On occasion I will prepare cross sections of embedded polymer fiber networks, polymer films and wood
fiber based structures. Would some of you who have used or maybe still use this tool give me some
recommendations on knives, where you prefer to have them sharpened and preferred knife orientation
for optimum cutting? I would appreciate it.

Tom Kremer
IPS Testing

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Email: dlowry-at-asu.edu
Name: David Lowry

Organization: Arizona State Univ

Title-Subject: [Filtered] need help identifying unknown bug

Message: we are seeking help from list-members with microbiology background in identifying an
endoparasite that is causing contamination problems in cultures of green algae. The contaminant
appears to be a bacterium but possesses an unusual morphology and apparent mode of reproduction. If
you might be able to assist, please contact me off-line and I can forward a few representative
images for evaluation. thank you-

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Email: maryard-at-uga.edu
Name: Mary Ard

Organization: University of Georgia, College of Veterinary Medicine

Title-Subject: [Filtered] Ultracut S control box shut down

Message: I have a Reichert Ultracut S ultramicrotome which has served me well for many years with
little to no problems until yesterday. The control box shut down completely. I have checked all
outside connections and have replaced the fuse (which I found blown - 220v). I'm sure it is an
internal problem, but I hesitate to open the box without an expert opinion. When replacing with
another 220v fuse, I can feel some activity coming from the inside, but the control box does not
come on. When replacing with a 110v fuse, the box hums, the initial light display comes on, but
then shuts down. Has anyone had a similar experience with this?

Regards,
Mary

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Email: jefbettini-at-gmail.com
Name: Jefferson Bettini

Organization: Brazilian Nanotechnology National Laboratory (LNNano)

Title-Subject: [Filtered] We are buying a Dual Beam FIB

Message: This laboratory is currently evaluating the purchase of a dual beam FIB and I recently
learned that Carlos Kazuo Inoki, who is an employee from this laboratory, posted some comments about
products from FEI, ZEISS and other companies, in internet sites. These comments represent
exclusively the personal opinion from Carlos Inoki and they do not express any institutional
assessment or opinion of the Brazilian Nanotechnology National Laboratory (LNNano), represented by
its management. Indeed, members of this laboratory have a completely different opinion from Carlos
Inoki.
LNNano management and personnel have been in contact with different companies concerning this
acquisition and we welcome their proposals since this will allow us to reach a sound decision,
considering the needs of our users and researchers. Finally, we acknowledge JEOL and FEI for all the
support these companies have provided to their microscopes currently installed in this laboratory.

Jefferson Bettini
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Name: Marissa

Organization: LBNL

Title-Subject: [Filtered] water-solvent swap

Message: I have nanoparticles in what I suspect to be dirty water and would like to transfer them
into a clean solvent for drop-casting. Is it possible to evaporate the water and then add the
appropriate solvent or am I completely oblivious to some well known nanoparticle in solution rule?

Thanks!

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Marissa,

Evaporating the water will only concentrate the impurities and will likely
aggregate the nano particles as well. You need to look at methods using
filtration or centrifugation. However, I would start by looking at the
method used to prepare the nano particles initially to see if there are
ways to prevent the contamination in the first place.

Debby

Debra Sherman, Chief Scientific Officer
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Hi Marissa,

There's a lot to consider when contemplating cleaning up nanoparticles
in solution. Do the nanoparticles have any form of functionalization?
A peptide group? A surfactant? Anything? or do they just have an
appropriate counterion in the solution which keeps them from
flocculating/agglomerating? The answer there will determine your next
step, and whether or no you can easily consider changing solvents.

Assuming your particles have something (surfactant or otherwise) to
prevent agglomeration, then you can consider a centrifugation / wash /
re-suspend / (sonication?) / centrifugation... routine.

Depending on the weight/size of your nanoparticles, you can also
consider using dialysis tubing, as long as there is a significant enough
weight difference between your impurity and your nanoparticles. That
will also determine the diffusing species and direction.

Once you get past those questions, then you can start to consider
whether or not you can even suspend the nanoparticles in your desired
solvent.

-John

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8, 21 -- From jpapalia-at-papalia.net Sat Jan 12 18:52:23 2013
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Email: rjh40-at-esc.cam.ac.uk
Name: Richard Harrison

Organization: University of Cambridge

Title-Subject: [Filtered] Post-Doctoral Research Associate in Focussed Ion Beam (FIB)/Electron
Microscopy, University of Cambridge

Message: Post-Doctoral Research Associate in Focussed Ion Beam (FIB)/Electron Microscopy

Applications are invited for a post-doctoral research associate (PDRA) position within the mineral
magnetism group of the Department of Earth Sciences, University of Cambridge. The position is funded
by the ERC grant Nanopaleomagnetism: a new multiscale approach to paleomagnetic analysis of
geological materials, a brief description of which is given below. Funding is available for 3 years
in the first instance. Successful applicants will hold a PhD on taking up the post.

The PDRA project will focus on the application of a dual-beam focussed ion beam (FIB) workstation to
perform 3D slice-and-view tomography of natural samples. The person appointed will use facilities
located in the Department of Materials Science to study the positions, sizes, shapes, compositions
and crystallographic orientations of magnetic particles embedded in silicate hosts using a
combination of slice-and-view tomography, electron backscattered diffraction (EBSD) and energy
dispersive X-ray analysis (EDX). We seek candidates with a strong background in electron microscopy
and specific experience with using FIB techniques. Experience with EDX, EBSD and tomography would be
an advantage. It is not necessary to have a background in either Earth sciences or magnetism, and we
invite suitably experienced candidates from any area of the physical sciences. The successful
candidate will have a track record of publication in peer-reviewed journals, will have good
communication skills and have demonstrated the capacity to perform independent research.

For full details see http://www.jobs.cam.ac.uk/job/-24782/

Interested candidates should contact Dr. Richard Harrison (rjh40-at-esc.cam.ac.uk) for more information
about the project and formal application procedures.

General description of the ERC project:

Adopting cutting-edge techniques from physics and materials science, Nanopaleomagnetism aims to
perform paleomagnetic measurements at submicron length scales, enabling primary magnetic signals to
be extracted from ancient and severely altered geological materials. 3D measurements of the volume,
shape and spacing of all magnetic particles within a microscale region of interest will be made
using a dual beam focussed ion beam workstation. Combined with high-resolution paleomagnetic
measurements and nanometre/nanosecond electron/X-ray magnetic imaging, nanopaleomagnetism will, for
the very first time, be able to characterise the magnetic properties of geological materials at
fundamental length scales and time scales. The nanoscale measurements will enable us to capture the
essential physics of the remanence acquisition process and to explore magnetic behaviour in
silico, allowing predictions to be made that can be tested directly against experimental
observations at all length scales. Sample-return missions to asteroids, comets, moons and planets
will soon provide unprecedented opportunities for extraterrestrial paleomagnetism.
Nanopaleomagnetism will provide the methodology and instrumentation needed to analyse these precious
materials.

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Dear Researchers:

I am doing a cryosubstitution run. The samples being processed need to
be positioned in a certain way for Lowicryl embedding. The process of
positioning samples takes time and needs to be done under a dissecting
scope, therefore it would be easier if it is done when the samples are at
a higher temperature (near zero), and subsequently embed/UV polymerize in
Lowicryl at the same temperature. Has anyone tried this? Is there
anything I need to be aware when UV polymerize Lowicryl at this
temperature? Thanks in advance.

Hong

________________________________

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Dear Researchers:

I would like to get some information about JEOL JEM-1400 TEM. If you own
this microscope, I would be delighted to hear from you off-line. Thank
you very much in advance.

Hong

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Hong,

I have often polymerized Lowacryl HM20 using UV light at 0oC or -20oC. In
fact, for years we used a makeshift UV setup in a refrigerator freezer. I
just cut out poster board and covered them with aluminum foil to make a
reflective chamber. I had two plexiglass rods that were poked through the
side pieces and supported a test tube rack. The bottom wires on the rack
were just the right size for suspending Beem capsules by their lids. It
also would hold flat molds if necessary. All the actual orientation was
done in the flat mold on a cold tray using a dissecting microscope.In that
case the mold on the cold tray could be moved to the reflective chamber
and UV light suspended from above.

Later we did get a Leica FS unit with the binocular attachment. This
meant that we could orient in flat molds while still at very low
temperatures and then not have to move the mold until after
polymerization. That works great so we could then polymerize at much
lower temperatures. I found that many antigens were preserved just fine
at 0 or -20 but then there are those elusive few where labeling really
seemed to benefit from lower temperatures.

Debby

Debra Sherman, Chief Scientific Officer
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

On 1/13/13 12:47 PM, "hyi-at-emory.edu" {hyi-at-emory.edu} wrote:

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Title-Subject: [Filtered] Porter Blum MT2 Ultramicrotome Manual/Parts?

Message: I bought a MT2, and, of course, I need parts and a manual(hard or electronic). Can anyone
suggest some sources?

Anyone using one these days? How do they still run?

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Hi, Hong,

Is it possible to do a partial orientation on the samples, that is, to orient them in one direction, and polymerize them in the Lowicryl at the usual temperature, then cut the polymerized blocks with a saw, and Super-Glue the cut blocks onto other blocks to obtain your final orientation for sectioning?

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: hyi-at-emory.edu [hyi-at-emory.edu]
Sent: Sunday, January 13, 2013 12:52 PM
To: Haller, Edward

Dear Researchers:

I am doing a cryosubstitution run. The samples being processed need to
be positioned in a certain way for Lowicryl embedding. The process of
positioning samples takes time and needs to be done under a dissecting
scope, therefore it would be easier if it is done when the samples are at
a higher temperature (near zero), and subsequently embed/UV polymerize in
Lowicryl at the same temperature. Has anyone tried this? Is there
anything I need to be aware when UV polymerize Lowicryl at this
temperature? Thanks in advance.

Hong

________________________________

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Dear All,

With Lowicryl, it is also possible to embed using UV illumination at low temperature without regard for orientation, and then re-embed the block in epoxy resin using heat polymerization. We have been doing this for years with Lowicryl HM20 polymerized at -50�C, re-embeding in epoxy resin at +60�C.

The sections have been probed with a wide variety of antibodies, which have all shown no detectable reduction in antigenicity (as compared with non-heat treated Lowicryl blocks).

Re-embedding in epoxy resin offers an excellent way of determining orientation at room temp. To keep the orientation during polymerization of epoxy resin, the Lowicryl blocks are first "glued" in place using only a small amount of resin in the flat embedding molds. Once the polymerization has been started by heating in the oven, fresh resin is added to the mold to make the correct block shape.

Another trick when working with Lowicryl and difficult-to-see embedded specimens is to "mark" the specimen location in the polymerized block (and also the perimeter of the resin block) with a small drop of nail varnish (polish?). The small drop of color can be used to locate the specimen after re-embedding in epoxy resin.

Sincerely,

Paul Webster, Ph.D.
House Research Institute
2100 W. 3rd St
Los Angeles
CA 90057, USA

-----Original Message-----
X-from: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
Sent: Mon 1/14/2013 5:29 AM
To: Webster, Paul

Hi, Hong,

Is it possible to do a partial orientation on the samples, that is, to orient them in one direction, and polymerize them in the Lowicryl at the usual temperature, then cut the polymerized blocks with a saw, and Super-Glue the cut blocks onto other blocks to obtain your final orientation for sectioning?

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: hyi-at-emory.edu [hyi-at-emory.edu]
Sent: Sunday, January 13, 2013 12:52 PM
To: Haller, Edward

Dear Researchers:

I am doing a cryosubstitution run. The samples being processed need to
be positioned in a certain way for Lowicryl embedding. The process of
positioning samples takes time and needs to be done under a dissecting
scope, therefore it would be easier if it is done when the samples are at
a higher temperature (near zero), and subsequently embed/UV polymerize in
Lowicryl at the same temperature. Has anyone tried this? Is there
anything I need to be aware when UV polymerize Lowicryl at this
temperature? Thanks in advance.

Hong

________________________________

This e-mail message (including any attachments) is for the sole use of
the intended recipient(s) and may contain confidential and privileged
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I agree with the sentiments of Paul and Debbie. I frequently embed my tissues in Lowicryl, LR Gold, and BMMA at 4 C and have excellent polymerization. I generally start with a flat-bottom BEEM capsule with a cap on it. I make no special effort to orient the tissue at the initial stage and Murphy's Law ensures that I get the wrong orientation even less often than chance might predict. I cut the blocks out of the BEEM capsule with a razor blade and inspect the surface with tissue in it under a stereoscope. I then mark the surface with a Sharpie and use a Dremel to trim away the plastic I don't want (i.e., where the Sharpie ink is). This generally results in a somewhat rectangle block that could be sectioned in a pinch but I usually insert this chunk of plastic back in a fresh BEEM capsule - the size of the block makes it easy to orient like I want and it can't easily shift in the capsule. I add a fresh label and resin and repolymerize. It costs me an extra day but I end up with a well-oriented block that is fully complete for inserting into the microtome chuck.

I find 4 C is sufficient for most antigens but if the immunolabeling doesn't work, I consider using a lower temperature embedding procedure. I list below three papers that discuss the effect of temperature on Lowicryl polymerization and/or immunolabeling. I also include a reference to a simple device I devised which allows tissues samples to be processed at sub-zero temperatures during dehydration, infiltration and polymerization. I built the device for less than $500 and it uses a conventional siphon-type carbon dioxide gas cylinder to maintain an aluminum block at temperatures as low as -35�C for over 15 hours/cylinder. Good luck. Tom

Armbruster BL, Garavito RM, Kellenberger E. 1983 Dehydration and embedding temperatures affect the antigenic specificity of tubulin and immunolabeling by the protein A-colloidal gold technique. J Histochem Cytochem 31(12):1380-1384.

Glauert A, Young RD. 1988. The control of temperature during polymerization of Lowicryl K4M: there is a low-temperature embedding method. J Microscopy 154(2):101-113.

Weibull C. 1987. Temperature rise in Lowicryl resins during polymerization by ultraviolet light. J Ultrastruct Molec Res 97:207-209.

Shoemaker, W., C. Hayes, and T.E. Phillips. 2003. A simple, low-cost device for processing and embedding tissues at sub-zero temperatures. Microscopy Research and Technique 62:262-266.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: PWebster-at-hei.org [mailto:PWebster-at-hei.org]
Sent: Monday, January 14, 2013 9:33 AM
To: Phillips, Thomas E.

Dear All,

With Lowicryl, it is also possible to embed using UV illumination at low temperature without regard for orientation, and then re-embed the block in epoxy resin using heat polymerization. We have been doing this for years with Lowicryl HM20 polymerized at -50�C, re-embeding in epoxy resin at +60�C.

The sections have been probed with a wide variety of antibodies, which have all shown no detectable reduction in antigenicity (as compared with non-heat treated Lowicryl blocks).

Re-embedding in epoxy resin offers an excellent way of determining orientation at room temp. To keep the orientation during polymerization of epoxy resin, the Lowicryl blocks are first "glued" in place using only a small amount of resin in the flat embedding molds. Once the polymerization has been started by heating in the oven, fresh resin is added to the mold to make the correct block shape.

Another trick when working with Lowicryl and difficult-to-see embedded specimens is to "mark" the specimen location in the polymerized block (and also the perimeter of the resin block) with a small drop of nail varnish (polish?). The small drop of color can be used to locate the specimen after re-embedding in epoxy resin.

Sincerely,

Paul Webster, Ph.D.
House Research Institute
2100 W. 3rd St
Los Angeles
CA 90057, USA

-----Original Message-----
X-from: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
Sent: Mon 1/14/2013 5:29 AM
To: Webster, Paul

Hi, Hong,

Is it possible to do a partial orientation on the samples, that is, to orient them in one direction, and polymerize them in the Lowicryl at the usual temperature, then cut the polymerized blocks with a saw, and Super-Glue the cut blocks onto other blocks to obtain your final orientation for sectioning?

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: hyi-at-emory.edu [hyi-at-emory.edu]
Sent: Sunday, January 13, 2013 12:52 PM
To: Haller, Edward

Dear Researchers:

I am doing a cryosubstitution run. The samples being processed need to be positioned in a certain way for Lowicryl embedding. The process of positioning samples takes time and needs to be done under a dissecting scope, therefore it would be easier if it is done when the samples are at a higher temperature (near zero), and subsequently embed/UV polymerize in Lowicryl at the same temperature. Has anyone tried this? Is there anything I need to be aware when UV polymerize Lowicryl at this temperature? Thanks in advance.

Hong

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Dear Collective,

Does anyone know of a source of flexible tubing for use with liquid nitrogen, outside diameter 6mm and inside diameter 4mm? Our cryoultramicrotome setup has tubing labeled as Festo PP-4, which the manufacturer says is rated down to -30 C and it cracks and breaks easily at LN2 temps. This dewar-to-cutting chamber hose can be easily repaired with the right tubing, but I'm not having any luck finding anything suitable in the right size.

Not optimistic, but eternally hopeful!

Cheers and thanks,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

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to: tindallr-at-missouri.edu

Good afternoon,
Dear Randy,
since here it is time to go home (8.40 p.m.) and your request was received some minute before leaving the lab hopefully very briefly:
my first thought was and my suggestion is (not very {inexpensive} I guess but most probably getting rid of broken hoses.)
I remember "old days" in the late 1970ies when I had a similar problem with tubing on an old cryo-ultramicrome...(Reichert OMU3 or was it an old Ultracut? with "cryochamber")
try Google Search {TEFLON} or PTFE tubing / tubes e.g. (no interest, no affiliation, just one out of the results found):

http://www.zeusinc.com/extrusionservices/materials/ptfe.aspx

PTFE Background Information
As the world's largest manufacturer of PTFE Tubing, Zeus offers a wide range of standard size products and precision custom PTFE Tubing. PTFE Tubing has become the gold standard in industries requiring the ultimate in lubricity, high temperature use, chemical resistance, biocompatibility, and precision extruded tolerances.
Key Properties
Very Lubricious - Lowest coefficient of friction of any polymer
Working temperature range 500� F (260� C) to -454� F (-270� C)
Chemically Resistant (all common solvents, acids and bases)
Chemically Inert
Low extractable
Excellent Dielectric Insulation Properties
For specific properties, see our Summary of Properties ....

and: http://www.ptfemart.com/ptfe-tubing.htm
it might be you'll find an appropriate out-/Inside diameter, for better thermic "shielding"(also not to get burned accidentally) one could wrap around foam-material ("thermo"-sleeving http://www.ptfemart.com/ptfe-tubing.htm material).

Good luck and best regards,

Wolfgang

Wolfgang MUSS
SALZBURG-AUSTRIA

==============================================

Von: tindallr-at-missouri.edu [mailto:tindallr-at-missouri.edu]
Gesendet: Montag, 14. J�nner 2013 20:18
An: Mu� Wolfgang
Betreff: [Microscopy] Cryo tubing?

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Dear Collective,

Does anyone know of a source of flexible tubing for use with liquid nitrogen, outside diameter 6mm and inside diameter 4mm?
Our cryoultramicrotome setup has tubing labeled as Festo PP-4, which the manufacturer says is rated down to -30 C and it cracks and breaks easily at LN2 temps.

This dewar-to-cutting chamber hose can be easily repaired with the right tubing, but I'm not having any luck finding anything suitable in the right size.

Not optimistic, but eternally hopeful!

Cheers and thanks,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

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Hey all, we've produced a few instructional videos here that go over how to use the Hitachi S-4800 and the S-3400. We've had some pretty good feedback on them and thought we would share them here in case anyone was interested.

Full 3400 instructions
http://youtu.be/BOUFKItQtHc

Quickie 3400 instructions
http://youtu.be/uQ8J2E3VzGk

Full 4800 instructions
http://youtu.be/52UKCY9fUR0

ERiC Jay Miller
Microscopy & Imaging Specialist
Electron Probe Instrumentation Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1152 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 467-0789
fax: (847) 467-657

http://www.nuance.northwestern.edu

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Rather than re-embedding, if tissue is not correctly oriented for sectioning after polymerisation, we usually cut off the tip of the block with the specimen in such a way that we can superglue it, with tissue in correct orientation, onto a piece of perspex rod that's the right size for the microtome chuck. We get our workshop to cut the rod into the correct lengths and polish the ends. We get them to make a couple of hundred each time, it doesn't take them long and there's usually an apprentice who gets the job...

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Tuesday, 15 January 2013 3:06 a.m.
To: White, Rosemary (PI, Black Mountain)

I agree with the sentiments of Paul and Debbie. I frequently embed my tissues in Lowicryl, LR Gold, and BMMA at 4 C and have excellent polymerization. I generally start with a flat-bottom BEEM capsule with a cap on it. I make no special effort to orient the tissue at the initial stage and Murphy's Law ensures that I get the wrong orientation even less often than chance might predict. I cut the blocks out of the BEEM capsule with a razor blade and inspect the surface with tissue in it under a stereoscope. I then mark the surface with a Sharpie and use a Dremel to trim away the plastic I don't want (i.e., where the Sharpie ink is). This generally results in a somewhat rectangle block that could be sectioned in a pinch but I usually insert this chunk of plastic back in a fresh BEEM capsule - the size of the block makes it easy to orient like I want and it can't easily shift in the capsule. I add a fresh label and resin and repolymerize. It costs me an extra day but I end up wi!
th a well-oriented block that is fully complete for inserting into the microtome chuck.

I find 4 C is sufficient for most antigens but if the immunolabeling doesn't work, I consider using a lower temperature embedding procedure. I list below three papers that discuss the effect of temperature on Lowicryl polymerization and/or immunolabeling. I also include a reference to a simple device I devised which allows tissues samples to be processed at sub-zero temperatures during dehydration, infiltration and polymerization. I built the device for less than $500 and it uses a conventional siphon-type carbon dioxide gas cylinder to maintain an aluminum block at temperatures as low as -35�C for over 15 hours/cylinder. Good luck. Tom

Armbruster BL, Garavito RM, Kellenberger E. 1983 Dehydration and embedding temperatures affect the antigenic specificity of tubulin and immunolabeling by the protein A-colloidal gold technique. J Histochem Cytochem 31(12):1380-1384.

Glauert A, Young RD. 1988. The control of temperature during polymerization of Lowicryl K4M: there is a low-temperature embedding method. J Microscopy 154(2):101-113.

Weibull C. 1987. Temperature rise in Lowicryl resins during polymerization by ultraviolet light. J Ultrastruct Molec Res 97:207-209.

Shoemaker, W., C. Hayes, and T.E. Phillips. 2003. A simple, low-cost device for processing and embedding tissues at sub-zero temperatures. Microscopy Research and Technique 62:262-266.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: PWebster-at-hei.org [mailto:PWebster-at-hei.org]
Sent: Monday, January 14, 2013 9:33 AM
To: Phillips, Thomas E.

Dear All,

With Lowicryl, it is also possible to embed using UV illumination at low temperature without regard for orientation, and then re-embed the block in epoxy resin using heat polymerization. We have been doing this for years with Lowicryl HM20 polymerized at -50�C, re-embeding in epoxy resin at +60�C.

The sections have been probed with a wide variety of antibodies, which have all shown no detectable reduction in antigenicity (as compared with non-heat treated Lowicryl blocks).

Re-embedding in epoxy resin offers an excellent way of determining orientation at room temp. To keep the orientation during polymerization of epoxy resin, the Lowicryl blocks are first "glued" in place using only a small amount of resin in the flat embedding molds. Once the polymerization has been started by heating in the oven, fresh resin is added to the mold to make the correct block shape.

Another trick when working with Lowicryl and difficult-to-see embedded specimens is to "mark" the specimen location in the polymerized block (and also the perimeter of the resin block) with a small drop of nail varnish (polish?). The small drop of color can be used to locate the specimen after re-embedding in epoxy resin.

Sincerely,

Paul Webster, Ph.D.
House Research Institute
2100 W. 3rd St
Los Angeles
CA 90057, USA

-----Original Message-----
X-from: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
Sent: Mon 1/14/2013 5:29 AM
To: Webster, Paul

Hi, Hong,

Is it possible to do a partial orientation on the samples, that is, to orient them in one direction, and polymerize them in the Lowicryl at the usual temperature, then cut the polymerized blocks with a saw, and Super-Glue the cut blocks onto other blocks to obtain your final orientation for sectioning?

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: hyi-at-emory.edu [hyi-at-emory.edu]
Sent: Sunday, January 13, 2013 12:52 PM
To: Haller, Edward

Dear Researchers:

I am doing a cryosubstitution run. The samples being processed need to be positioned in a certain way for Lowicryl embedding. The process of positioning samples takes time and needs to be done under a dissecting scope, therefore it would be easier if it is done when the samples are at a higher temperature (near zero), and subsequently embed/UV polymerize in Lowicryl at the same temperature. Has anyone tried this? Is there anything I need to be aware when UV polymerize Lowicryl at this temperature? Thanks in advance.

Hong

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Email: rleepenn-at-hotmail.com
Name: Lee Penn

Title-Subject: [Filtered] water-solvent swap

Message: In response to: Message: I have nanoparticles in what I suspect to be dirty water and
would like to transfer them into a clean solvent for drop-casting. Is it possible to evaporate the
water and then add the appropriate solvent or am I completely oblivious to some well known
nanoparticle in solution rule?
Thanks!

No - drying could change your particles irreversibly and cause substantial aggregation.

I recommend using dialysis against purified water.
If the things you want to remove from the suspension are dissolved, then the dialysis will result in
the diffusion of the dissolved species from the suspension to the surrounding purified water.

Check out the experimental section of this paper, and you will find details about how to perform
dialysis against purified water in order to remove dissolved species from a suspension of
nanoparticles and water.
Effect of Ionic Strength on the Kinetics of Crystal Growth by Oriented Aggregation, Nathan D.
Burrows, Christopher R. H. Hale, and R. Lee Penn (2012) Crystal Growth and Design, DOI:
10.1021/cg3004849 Article ASAP.

If you have a few more details about your nanoparticles, I'd be happy to chat about sample prep
options. We have worked with a range of nanoparticles in aqueous systems, ranging from very "dirty"
suspensions of natural materials in natural waters to nanoparticles harvested from aqueous reactors
to very clean systems that have been dialyzed against purified water.

Good luck and happy new year!

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Organization: Drexel University

Title-Subject: [Filtered] Reading beam current on ESEM Philips FEI XL30

Message: Hello all,

I am using a Philips FEI XL30 ESEM machine. I was wondering if it was possible to...
(1) measure the current that is striking the SAMPLE (not the filament).
(2) control the intensity of the beam current aside from changing the spot size (eg, specify a beam
with current of 1nA instead of spotsize 1)

My ultimate goal is to attain a beam with a current on the order of nano amps.

Any input will be appreciated,

Thank you
Gqiu

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Remembering back to when we did e-beam lithography on that microscope, there was a BNC connector on the door. We would plug our ammeter into this connector, and it seems that we had to disconnect a wire that grounded the stage inside the chamber. Disconnecting the wire would disable the touch sensor, so we had to be extra careful about not crashing the pole piece.

Jonathan Abbott

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} Title-Subject: [Filtered] Reading beam current on ESEM Philips FEI XL30
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} I am using a Philips FEI XL30 ESEM machine. I was wondering if it was possible to...
} (1) measure the current that is striking the SAMPLE (not the filament).
} (2) control the intensity of the beam current aside from changing the spot size (eg, specify a beam
} with current of 1nA instead of spotsize 1)
}
} My ultimate goal is to attain a beam with a current on the order of nano amps.
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} Gqiu
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{Note that this is a reply to a web question. I copied them as well as the list server.}

Your request is quite normal, except for the part about doing it without changing the "spot size".

You will probably need a Faraday cup and a nano-ammeter to measure the current reaching your sample. I would like to think you could find such a meter without much trouble at Drexel. I don't know where you plug that into an XL30. I suppose it is on the front door, but others could say for sure. A Faraday cup may be as simple as small, deep hole drilled into a piece of carbon. If you want to get fancy, you can glue a used aperture over the top so that the electrons that enter the hole stay in the hole and get absorbed. In that case, sample current = beam current.

Now about "spot size" - I learned on SEMs that didn't have a "spot size" setting. They had condenser lens controls without any particular numbering. You adjusted the lens one way to choke the beam down for more resolution and the other way for more beam current. Of course, one of the side effects was that the size of the beam spot on the sample changed a little. I usually didn't worry about how much since I was working at lower magnifications (by today's standards). I choked it down for high resolution microscopy while maintaining a decent signal-to-noise ratio in my iamge. I increased it as necessary to get enough current for a decent count rate for x-ray analysis. The setting varied some depending on my needs and how much time I had. Of course, you can also change the objective aperture to minimize spot size for a given beam current. You'll have to experiment a little to see what gives you the sharpest image at 1 nA.

So again, what is your concern about the spot size? The three SEMs I work with now have spot sizes that range up to 7, 60, or 255. They are quite arbitrary and non-linear in their numbering. You should probably go through an exercise to see what your numbers correlate to in terms of incident beam size. I think your beam diameter will probably be no more than a few nm even at hefty currents. Hopefully you can focus better than many of our users so that you really do attain the optimum effect of that spot size.

My work tends to concentrate on microanalysis and the beam diameter is much smaller than the interaction volume diameter. I can safely ignore the beam diameter and just call it zero. Interaction volume is the main determinant of x-ray resolution. So now I spend a fair amount of time determining the interaction volume as a function of voltage in an effort to keep my interaction within the phase of interest while still exciting lines that I can resolve in the EDS spectra.

Of course, you may have a different issue at hand, maybe e-beam lithography, but I think you will still find that the interaction volume is more the determining factor. Set the condenser lens (spot size) where you need to at the voltage you need to.

Warren Straszheim
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Title-Subject: [Filtered] Reading beam current on ESEM Philips FEI XL30

Message: Hello all,

I am using a Philips FEI XL30 ESEM machine. I was wondering if it was possible to...
(1) measure the current that is striking the SAMPLE (not the filament).
(2) control the intensity of the beam current aside from changing the spot size (eg, specify a beam
with current of 1nA instead of spotsize 1)

My ultimate goal is to attain a beam with a current on the order of nano amps.

Any input will be appreciated,

Thank you
Gqiu

Login Host: 144.118.157.21
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18, 37 -- From: "Straszheim, Warren E [BIOTC]" {wesaia-at-iastate.edu}
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18, 37 -- Subject: RE: [Microscopy] viaWWW:Reading beam current on ESEM Philips FEI
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Hi

You have had a good deal of advice on this problem but may I address the
control of the beam current? There are three variables

1) Emission current from the gun - bias setting or filament position -
move the filament toward the cathode aperture for a higher current or
increase the emission current control.

2) The setting of the first condenser lens - stronger lens for a lower
current.

3) The size of the beam defining aperture - larger aperture for more
current.

X-from my knowledge there are hardly any, if any instruments that do directly
link the beam current with control of the condenser system. Whilst the
manufacturers will often guess the current they will be placing on the
specimen (their readout figure) this guess is only true for their idea of
where the above three variables are set; their guess is very much an
approximation! The only true indication of beam current is the Faraday Cup!

Enjoy

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Tel & Fax +44 (0)1280 816512 Cell +44 (0)7711 606967
www.emcourses.com

REPLY TO

Email: gangqiu0-at-gmail.com
Name: Gqiu

Organization: Drexel University

Title-Subject: [Filtered] Reading beam current on ESEM Philips FEI XL30

Message: Hello all,

I am using a Philips FEI XL30 ESEM machine. I was wondering if it was
possible to...
(1) measure the current that is striking the SAMPLE (not the filament).
(2) control the intensity of the beam current aside from changing the spot
size (eg, specify a beam with current of 1nA instead of spotsize 1)

My ultimate goal is to attain a beam with a current on the order of nano
amps.

Any input will be appreciated,

Thank you
Gqiu

==============================Original Headers==============================
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23, 23 -- Subject: RE: [Microscopy] viaWWW:Reading beam current on ESEM Philips FEI XL30
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Title-Subject: [Filtered] Lehigh Microscopy School

Message: There is still time to register for the 2013 Lehigh Microscopy School which will be held
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will be the 43rd anniversary for the Lehigh Microscopy School and course offerings will include:
Introduction to SEM and EDS for the New Operator SEM and X-ray Microanalysis Focused Ion
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Message: The South Carolina EPSCoR/IDeA Program and the USC School of Medicine Instrumentation
Resource Facility are pleased to announce the 9th annual workshop on Basic Confocal Microscopy. The
Workshop will be held June 10-14 at the USC School of Medicine in Columbia, SC.

Workshop material is directed towards beginning and intermediate users of confocal microscopes and
involves a series of lectures (specimen preparation, labeling strategies, proper set-up of
instrument operating parameters, proper handling of 2D and 3D confocal images in programs such as
Adobe Photoshop and AMIRA), hands on specimen preparation, and time on a number of different point
scanning and spinning disk confocal microscopes.

Companies that have provided equipment and applications experts for past workshops include Leica,
Nikon, Olympus, Perkin Elmer, Zeiss, Intelligent Imaging Innovations (3i) and Photometrix.
Participants will have ample time for hands on use of the instruments during the workshop

Faculty will include Dr. Jay Jerome of Vanderbilt University, Dr. Ralph Albrecht of the University
of Wisconsin Madison, Dr. John Mackenzie of North Carolina State University, Dr. Tom Trusk of the
Medical University of South Carolina, and Dr. Bob Price of the University of South Carolina.

For further information please see: http://dba.med.sc.edu/price/irf/irf.htm or contact Anna McNeal
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Title-Subject: [Filtered] Post-doc available in electron microscopy

Message: Quantum Non-Demolition Electron Microscope (QEM) Post-doctoral candidates are sought with
experience in one or more of: quantum optics, free-electron optics design, numerical simulation of
electron systems, electron decohe rence, and/or transmission-electron microscopy. Each individual
will work with a small international team of researchers at MIT (K. Berggren and M. F. Yanik) and
the Technical University of Delft (P. Kruit) in the Netherlands, Stanford University (M. Kasevich),
and Max Planck Institute of Quantum Optics (P. Hommelhoff) to develop a new electron mi croscope
based on quantum non-demolition measurements (Putnam & Yanik, Phys. Rev. A 80, 040902 (R) 2009). The
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Email: Uchechukwu_Mba-at-yahoo.com
Name: Uchechukwu Mba

Organization: نٍ شيَا خُوِ
ذُوْ ذِ جِ ثُوْ
宁夏大学

Title-Subject: [Filtered] SEM chamber contamination

Message: Dear List members,

Our laboratory recently purchased a specially commissioned SEM. The idea was that instrument was
as oil free as possible, so it was equipped with a TMP and a scroll pump. To our dismay, there is a
substantial contamination: the clearly visible infamous dark squares. They are currently
negotiating with the manufacturer but in the mean time we need to assess its quantity and type.
We are thinking of preparing some polished sample and collect a set of images at different
integration times and make a plot of darkness vs. time. The question is however, what would be an
acceptable darkening rate?

Does anyone know if there exists some sort of standardized protocol?

Can anyone recommend a procedure?

Some recommendations in dealing with the manufacturer are also welcome. Their first reaction was
It is from your samples!!!. Fortunately, it was very easy to show this was not the case. Now they
are improvising all sorts of cleanings.

Thanks a lot
Mba

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Hi Mba,
You should be able to scan an area for several minutes, without seeing
build-up. See the NIST paper (Evactron.com).

Short of a UHV solution, it is not surprising to have contamination in a SEM
or FIB. There are so many HC's diffusing out of crevices and off surfaces
from machined parts and plastics in the instrument, especially when it is
newer. If you have budget, an in-situ cleaning device like the GV-10x or
Evactron is extremely helpful.

Regards,
Larry Scipioni
ZS Genetics

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Email: Uchechukwu_Mba-at-yahoo.com
Name: Uchechukwu Mba

Organization: نٍ شيَا
خُوِ ذُوْ ذِ
جِ ثُوْ 宁夏大学

Title-Subject: [Filtered] SEM chamber contamination

Message: Dear List members,

Our laboratory recently purchased a specially commissioned SEM. The idea
was that instrument was as oil free as possible, so it was equipped with a
TMP and a scroll pump. To our dismay, there is a substantial contamination:
the clearly visible infamous dark squares. They are currently
negotiating with the manufacturer but in the mean time we need to assess its
quantity and type.
We are thinking of preparing some polished sample and collect a set of
images at different integration times and make a plot of darkness vs. time.
The question is however, what would be an acceptable darkening rate?

Does anyone know if there exists some sort of standardized protocol?

Can anyone recommend a procedure?

Some recommendations in dealing with the manufacturer are also welcome.
Their first reaction was It is from your samples!!!. Fortunately, it was
very easy to show this was not the case. Now they are improvising all sorts
of cleanings.

Thanks a lot
Mba

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28, 22 -- From les-at-zsgenetics.com Wed Jan 16 08:56:26 2013
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Contents Retrieved from Microscopy Listserver Archives
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For a protocol you might look at:

Conru and LaBerge, Oil Contamination with SEM Operated in Spot Scan Mode. J
Phys E Sci Instr 8(2): 136-138, 1975.

Tony

��
Andrew Anthony �Tony� Havics, CHMM, CIH, PE
Environmental, Health & Safety, Microscopy, & Forensic Engineering
pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
(317) 718-7020 Office
(317) 718-7038 Fax
(317) 409-3238 Cell
aahavics-at-pH2LLC.com
www.ph2LLC.com

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Name: Uchechukwu Mba

Organization: نٍ شيَا
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ثُوْ
宁夏大学

Title-Subject: [Filtered] SEM chamber contamination

Message: Dear List members,

Our laboratory recently purchased a specially commissioned SEM. The idea
was that instrument was
as oil free as possible, so it was equipped with a TMP and a scroll pump. To
our dismay, there is a
substantial contamination: the clearly visible infamous dark squares.
They are currently
negotiating with the manufacturer but in the mean time we need to assess its
quantity and type.
We are thinking of preparing some polished sample and collect a set of
images at different
integration times and make a plot of darkness vs. time. The question is
however, what would be an
acceptable darkening rate?

Does anyone know if there exists some sort of standardized protocol?

Can anyone recommend a procedure?

Some recommendations in dealing with the manufacturer are also welcome.
Their first reaction was
It is from your samples!!!. Fortunately, it was very easy to show this was
not the case. Now they
are improvising all sorts of cleanings.

Thanks a lot
Mba

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From noreply-at-onlinebrusselsguide.com Wed Jan 16 10:17:51 2013
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Mba

Do contact XEI Scientific Inc., in Redwood, Ca. They sell devices which plug
into SEMs to decontaminate them of hydrocarbons. You can find them at
http://www.evactron.com/

Best Regards

Ian

Ian Holton
Acutance Scientific

-----Original Message-----
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To: ian-at-acutance.co.uk

For a protocol you might look at:

Conru and LaBerge, Oil Contamination with SEM Operated in Spot Scan Mode. J
Phys E Sci Instr 8(2): 136-138, 1975.

Tony

��
Andrew Anthony �Tony� Havics, CHMM, CIH, PE Environmental, Health & Safety,
Microscopy, & Forensic Engineering pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
(317) 718-7020 Office
(317) 718-7038 Fax
(317) 409-3238 Cell
aahavics-at-pH2LLC.com
www.ph2LLC.com

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Organization: نٍ شيَا
خُوِ
ذُوْ ذِ جِ
ثُوْ
宁夏大学

Title-Subject: [Filtered] SEM chamber contamination

Message: Dear List members,

Our laboratory recently purchased a specially commissioned SEM. The idea
was that instrument was
as oil free as possible, so it was equipped with a TMP and a scroll pump. To
our dismay, there is a
substantial contamination: the clearly visible infamous dark squares.
They are currently
negotiating with the manufacturer but in the mean time we need to assess its
quantity and type.
We are thinking of preparing some polished sample and collect a set of
images at different
integration times and make a plot of darkness vs. time. The question is
however, what would be an
acceptable darkening rate?

Does anyone know if there exists some sort of standardized protocol?

Can anyone recommend a procedure?

Some recommendations in dealing with the manufacturer are also welcome.
Their first reaction was
It is from your samples!!!. Fortunately, it was very easy to show this was
not the case. Now they
are improvising all sorts of cleanings.

Thanks a lot
Mba

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Contents Retrieved from Microscopy Listserver Archives
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What is the 'dirt'?
If it sediments then decant the less dirty supernate.
If what's decanted has dirt bigger than the nano-s, then filter - with forethought.
If the filtrate has dissolved material then use dialysis that won't pass the nano-s to reduce the volume wholistically and simultaneously concentrating the nano-s.
You might also use differential centrifugation.

All of the above depends on the size of the nano-s, and how they behave, AND whether you actually know they are present.

Hope this helps,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

HomePage: http://cmirt.wcupa.edu (with link to instrument Scheduler)

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Organization: LBNL

Title-Subject: [Filtered] water-solvent swap

Message: I have nanoparticles in what I suspect to be dirty water and would like to transfer them into a clean solvent for drop-casting. Is it possible to evaporate the water and then add the appropriate solvent or am I completely oblivious to some well known nanoparticle in solution rule?

Thanks!

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Mba,
I remember reading something in an old Kurt Lesker catalogue stating that if
you use a Turbo pump like a diffusion pump, with isolation valving so that
the pump can run continuously, it will be very clean. On the other hand, if
you run it the way many do, stopping it and bringing it to atmosphere every
time you vent the chamber, you will get oils creeping up through the turbo
pump and contaminating the chamber. This explained to me why SEMs that I
worked on where the DP had been replaced with a turbo were very clean and
Amrays (in particular) that had turbos and a single vent valve were so
terribly dirty.

Is your turbo in a system with roughing, backing and high vacuum isolation
valves, or just a single vent valve? If your turbo is mag-lev, then it is
most likely isolated and runs all the time due to the limited life of the
crash bearings. Otherwise, you might have the "simple" system, and it will
most likely be dirty.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

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Name: Uchechukwu Mba

Organization: نٍ شيَا
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宁夏大学

Title-Subject: [Filtered] SEM chamber contamination

Message: Dear List members,

Our laboratory recently purchased a specially commissioned SEM. The idea
was that instrument was
as oil free as possible, so it was equipped with a TMP and a scroll pump. To
our dismay, there is a
substantial contamination: the clearly visible infamous dark squares.
They are currently
negotiating with the manufacturer but in the mean time we need to assess its
quantity and type.
We are thinking of preparing some polished sample and collect a set of
images at different
integration times and make a plot of darkness vs. time. The question is
however, what would be an
acceptable darkening rate?

Does anyone know if there exists some sort of standardized protocol?

Can anyone recommend a procedure?

Some recommendations in dealing with the manufacturer are also welcome.
Their first reaction was
It is from your samples!!!. Fortunately, it was very easy to show this was
not the case. Now they
are improvising all sorts of cleanings.

Thanks a lot
Mba

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I believe that there is a basic limiting factor and that is electron current
density. This is pretty much determined by the source, and with thermionic
sources, when you increase the emission current you increase the area from
which electrons are emitted, all else being equal, you have a larger spot.

When you increase your condenser lens current, you get a smaller spot, but
with many fewer electrons. The same is true when you change final aperture
size.

In the end, there will be essentially a fixed spot size for a given beam
current when everything is focused. If the spot size you want is larger
than the nominal spot size for the current you need, you can just defocus
the beam. If you need a smaller spot for a given current, you need a
different electron source that will yield a higher current density.

Given all the variables in setting up a beam (kV, emission current,
condenser current, "objective" current, working distance, aperture sizes),
the only way to have a known current is to measure it using a Faraday cup
and pico-ammeter, as explained in other posts. You may be able to establish
some "standard settings" that will readily put you in the ball park, but if
the current is critical, it must be measured. Anyone doing quantitative
x-ray analysis or small scale ebeam lithography will confirm this.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: Tuesday, January 15, 2013 6:17 AM
To: kenconverse-at-qualityimages.biz

Hi

You have had a good deal of advice on this problem but may I address the
control of the beam current? There are three variables

1) Emission current from the gun - bias setting or filament position -
move the filament toward the cathode aperture for a higher current or
increase the emission current control.

2) The setting of the first condenser lens - stronger lens for a lower
current.

3) The size of the beam defining aperture - larger aperture for more
current.

X-from my knowledge there are hardly any, if any instruments that do
directly
link the beam current with control of the condenser system. Whilst the
manufacturers will often guess the current they will be placing on the
specimen (their readout figure) this guess is only true for their idea of
where the above three variables are set; their guess is very much an
approximation! The only true indication of beam current is the Faraday Cup!

Enjoy

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Tel & Fax +44 (0)1280 816512 Cell +44 (0)7711 606967
www.emcourses.com

REPLY TO

Email: gangqiu0-at-gmail.com
Name: Gqiu

Organization: Drexel University

Title-Subject: [Filtered] Reading beam current on ESEM Philips FEI XL30

Message: Hello all,

I am using a Philips FEI XL30 ESEM machine. I was wondering if it was
possible to...
(1) measure the current that is striking the SAMPLE (not the filament).
(2) control the intensity of the beam current aside from changing the spot
size (eg, specify a beam with current of 1nA instead of spotsize 1)

My ultimate goal is to attain a beam with a current on the order of nano
amps.

Any input will be appreciated,

Thank you
Gqiu

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Dear All,
today I mounted a new Denka LaB6 cathode first time and after firing it up got double contours on the image details.
Can anybody give me a hint if there is some basic problem with the geometry in the Wehnelt, e.g. cathode too far away or too near?
I mounted it as I normally mount Kimball LaB6 cathodes, centered very carefully, with a distance of the tip of ca. 0.15mm from the
UPPER (facing the Anode) Wehnelt plane.
It seems like there are two sources of electrons going out of the crystall. It might also come from the more "brick-like" shape of
the LaB6 cristal, in comparison with the round shape of a Kimball tip... When I heat the cathode up some notches more (I am now
nearly at the end of the scale), it`s still the same. I can post a link with images tomorrow, but today I missed taking some...
I put the same cathode in my Philips TEM EM420 and the image at 80 KV is perfect, also the cathode image during heating-up is as
it should be (in the TEM...).
Shall I first try and go nearer with the Wehnelt? Current at 30KV and medium Bias setting is now ca. 40 uA.

Any ideas?

Thanks,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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Email: gregc-at-physics.ucsb.edu
Name: Greg Courville

Organization: University of California, Santa Barbara

Title-Subject: [Filtered] ISI-SS40 documentation?

Message: Hello all,

We've just acquired an ISI-SS40 scanning electron microscope, with no documentation other than some
photocopied instructions and a manual for a different model (SX40). We contacted the service company
whose sticker we found on the unit, but they don't even have the docs for this model around any
more. If anyone has any documentation on this unit (in particular, schematics and/or a service
manual) that they wouldn't mind parting with or scanning, we'd love to hear from you and would be
willing to reimburse you as appropriate. Any other information on this series (we understand the
SS-60 and SS-160 are substantially similar instruments) would be greatly appreciated as well.

Best regards
--
Greg Courville

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Email: mlibbee-at-gmail.com
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Title-Subject: [Filtered] Graphene/scotch tape preparation

Message: Hello again. Is someone willing to share or recommend literature on a method for making a
graphene TEM sample using the scotch tape trick? Does this film need washing (HCl sol?) or plasma
cleaning to guarantee the adhesive is fully removed?

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***************************************************************************************
Forwarded from "Ask a Microscopist"
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****************************************************************************************
realname - Manuela Anstoetz
Email - manuela.anstoetz-at-scu.edu.au
ORGANIZATION - Southern Cross University
EDUCATION - Graduate College
LOCATION - Lismore, NSW, Australia
SUBJECT_OF_QUESTION - HRSEM and Ultra-HRSEM
QUESTION - Hello,
I would like to know if for HRSEM and Ultra-HRSEM Microscopy the premise
of low accelerating voltage gives more surface information and high
accelerating voltage delivers more sub-surface information is cancelled?
I have seen 30 kV being used for surface topology of a metallic whisker
growing from a thin film and feel confused about that. I looked up the
Microscope used (Hitachi SU5500) and discovered that it has a range for
Vacc from 0.5 keV to 30keV. I would have used 5keV or so to determine
surface morphology. Can you help clarify, please?
Thank you kindly,
Manuela

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Hi Manuela

First let me say that unfortunately not all the SEM results that you see in
publications are indicative of good microscopy! So much of what I see
suggests to me that the editors accept publications from those who know very
little about SEM! Now let's get down to your question.

No matter which SEM you use, no matter how clever its imaging system, the
basic physics of signal generation apply.

a) Secondary electrons are liberated from near the surface of the
material no matter what accelerating voltage is used.
b) Backscattered electrons are liberated from a volume below the
surface, the higher the accelerating voltage the deeper the depth from which
they may be generated.
c) Backscattered electrons striking parts of the chamber also produce
these signals. The BSE being converted into multiple secondary electrons
that may then contribute to the image. These converted BSE may be very
useful, are less effected by charge, therefore they often make a difficult
specimen usable. The problem is the converted BSE still carry the BSE data,
so the SE collected actually depict the sub surface information.

So to answer your question the lower the accelerating voltage the better if
you wish to see the true surface of a specimen. Lowering the accelerating
voltage reduces the depth of penetration allowing the SE signal to dominate.
Raising the accelerating voltage creates a larger volume from which BSE may
be produced. In this case the BSE themselves, or the converted BSE, may
dominate the information subduing the surface information.

You correctly note that the Hitachi have provided you with a very wide range
of accelerating voltages indicating that there is a very difficult and
critical decision to be made when you select your operating voltage.
Unfortunately 85% of operators around the world use the same accelerating
voltage no matter what type of specimen they are viewing. As a guide if you
wish to see the true surface of a specimen, you are correct, you should
operate at less than 5kV. However in a number of applications we may use
the collection of signals from different depths by changing the accelerating
voltage, "sectioning by kV", this technique often provides a mass of useful
information.

TTL(through the lens) detection is a very efficient method for collecting
SE, but if too many of the converted BSE are excluded charge may become a
major problem. Thus most knowledgeable operators will vary the WD with a
TTL system to vary the amount of converted BSE used to subdue charge. The
alternative is to work with a TTL system at very low accelerating voltages,
certainly less than 2kV, and to rely upon the superior collection efficiency
of the system to allow small spot sizes to be used, to minimise the probe
current thus minimise the charge. I am often working with clients at 1 to
1.5kV up to 150,000X in this type of system. We do not need to coat if we
carefully balance accelerating voltage with WD to make the impossible
possible.

Remember, there will be a particular accelerating voltage that will provide
more information than any other. How do we find this voltage? Well
microscopists are scientists and a scientist are meant to experiment!

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Tel & Fax +44 (0)1280 816512 Cell +44 (0)7711 606967
www.emcourses.com

****************************************************************************
************
realname - Manuela Anstoetz
Email - manuela.anstoetz-at-scu.edu.au
ORGANIZATION - Southern Cross University EDUCATION - Graduate College
LOCATION - Lismore, NSW, Australia SUBJECT_OF_QUESTION - HRSEM and
Ultra-HRSEM QUESTION - Hello, I would like to know if for HRSEM and
Ultra-HRSEM Microscopy the premise of low accelerating voltage gives more
surface information and high accelerating voltage delivers more sub-surface
information is cancelled?
I have seen 30 kV being used for surface topology of a metallic whisker
growing from a thin film and feel confused about that. I looked up the
Microscope used (Hitachi SU5500) and discovered that it has a range for Vacc
from 0.5 keV to 30keV. I would have used 5keV or so to determine surface
morphology. Can you help clarify, please?
Thank you kindly,
Manuela

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Manuela,

I agree entirely with Steve's comments below, but want to extend the
conversation a bit in a slightly different direction. You are the perfect
example of the importance of not just teaching but "education" to make a
good microscopist.

The easy way out is to sit down at a microscope and have someone show you
what knobs to turn or buttons to push or screen functions to activate to
get an image. It is also the fastest way to get users for your
instruments. Some of these users will question and gradually educate
themselves, but the majority will not. However, in my opinion, this is a
real disservice to not only to you, the user, but to others that use the
instrumentation for two reasons:

1) As Steve so ably explained below, you need to understand enough about
the principles behind SEM (and other forms of microscopy) so as to use the
systems to your advantage "in an educated way" to get the most out of the
instrument and your sample. Understanding the hows and whys of basic
operation will make you able to adapt not only to different samples and
the information needed from that sample, but also to different microscopes
as you advance your career. A good solid course in microscopy principles
is not a waste of time. It actually will speed up good data accumulation
down the line with more in-depth understanding of your samples in the
process.

2) Understanding how an instrument works and recognizing when a system is
not working to its full potential will help not only you but all the other
users of the instrument. You will often be able to recognize potential
problems, alert the appropriate people, and thus get the problems resolved
before they become serious. Also, responsible use of major equipment goes
hand-in-hand with understanding the system and also your limitations in
dealing with that system, so that you do not unknowingly or
unintentionally compromise the instrument.

It comes down to EDUCATION not just training if we want to develop, or be,
more than just instrument technicians, but truly competent microscopists.
Unfortunately microscopy courses are to often eliminated or diluted down
due to pressures to produce NOW not after completing a course as well as
less emphasis on education by faculty/supervisors/facility directors &
managers as they are confronted with these pressures.

Debby

Debra Sherman, Founder and CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

On 1/20/13 5:45 AM, "protrain-at-emcourses.com" {protrain-at-emcourses.com}
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17, 37 -- To: "protrain-at-emcourses.com" {protrain-at-emcourses.com} ,
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17, 37 -- "manuela.anstoetz-at-scu.edu.au"
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17, 37 -- Subject: Re: [Microscopy] RE: Ask-A-Microscopist HR SEM, kV, and surface
17, 37 -- topography-education!
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17, 37 -- topography-education!
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Hi

Back again to add to the comments Debby made.

A good friend of mine from Australia has this headline in his laboratory "Do
YOU do science or do YOU do tradition." Think about it?

Steve

-----Original Message-----
X-from: Sherman, Debra [mailto:dsherman-at-purdue.edu]
Sent: 20 January 2013 18:33
To: protrain-at-emcourses.com; message to: MSA list;
manuela.anstoetz-at-scu.edu.au

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To amplify what Debbie and Steve are saying:

I have always loved SEMs because they are a signal generator and a signal
processor, not a microscope. If you tell me what you want to see, I can
probably show it to you, then we need to have a long talk about what is
real. (What, you found argon in another sample?).

The world NEEDS microscopists! Anyone can take a pretty picture with the
new instruments, but are you sure of what the image is showing you?

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

-----Original Message-----
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Sent: Sunday, January 20, 2013 2:26 PM
To: kenconverse-at-qualityimages.biz

Hi

Back again to add to the comments Debby made.

A good friend of mine from Australia has this headline in his laboratory "Do
YOU do science or do YOU do tradition." Think about it?

Steve

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Marissa Libbee asked about "nanoparticles in [suspected] dirty water". I
don't have an answer to her question but want to share my own findings and
ask others for comments.

I'm studying nanoparticles in aqueous solution. The particles are in the
range 1-5 nm diameter. My task is to image them by AFM and to measure the
height of many individual particles. The sample preparation procedure is
basically to apply a droplet of the suspension to freshly cleaved mica,
rinse and dry. The AFM images are made in air at room temperature using
TappingMode (intermittent contact) and obtain nice images. However, blank
samples prepared from the ultrapure water used as diluent and as rinse water
also show some particles.
I have tried more than 3 different samples of ultrapure water produced using
Milli-Q water purification systems at 3 different labs and gotten similar
results.
I wonder whether anyone else has had similar experiences working with water
and mica.

regards,
Don
=============================================
Don Chernoff, Ph.D., President
Advanced Surface Microscopy, Inc.; 3250 N. Post Rd., Suite 120;
Indianapolis IN 46226 USA
E-Mail: donc-at-asmicro.com www.asmicro.com
Voice: 317-895-5630 Toll free: 800-374-8557 (in USA & Canada)
Fax: 317-895-5652
[business activities since 1990: analytical services in AFM, AFM probes,
consulting, training, calibration and test specimens,
calibration and measurement software, used NanoScope equipment.]
=============================================
----- Original Message -----
From: FMonson-at-wcupa.edu
To: donc-at-asmicro.com
Sent: Wednesday, January 16, 2013 12:42 PM
Subject: [a] [Microscopy] RE: viaWWW:water-solvent swap

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What is the 'dirt'?
If it sediments then decant the less dirty supernate.
If what's decanted has dirt bigger than the nano-s, then filter - with
forethought.
If the filtrate has dissolved material then use dialysis that won't pass
the nano-s to reduce the volume wholistically and simultaneously
concentrating the nano-s.
You might also use differential centrifugation.

All of the above depends on the size of the nano-s, and how they behave,
AND whether you actually know they are present.

Hope this helps,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

HomePage: http://cmirt.wcupa.edu (with link to instrument Scheduler)

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Subject: [Microscopy] viaWWW:water-solvent swap

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Email: mlibbee-at-gmail.com
Name: Marissa

Organization: LBNL

Title-Subject: [Filtered] water-solvent swap

Message: I have nanoparticles in what I suspect to be dirty water and
would like to transfer them into a clean solvent for drop-casting. Is it
possible to evaporate the water and then add the appropriate solvent or am I
completely oblivious to some well known nanoparticle in solution rule?

Thanks!

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Email: mzugravu-at-nesmicroscopy.org
Name: Monica Viorica Zugravu

Organization: Harvard University, Center for Nanoscale Systems

Title-Subject: [Filtered] NESM February, 28th Meeting -at- Boston University

Message: Greetings Fellow Microscopists,

This is your friendly Save the Date reminder for the NESM February Meeting hosted by Boston
University on February 28. The meeting will consist of a BU Photonics Center tour, a buffet dinner,
and two technical talks. Stay tuned for detailed information about the February Meeting in the
coming weeks. You can also keep up with NESM activities by checking us out on the web at
nesmicroscopy.org and on Facebook (https://www.facebook.com/NESMicroscopy) and Twitter
(https://twitter.com/#!/NESMicroscopy).

Cheers,
NESM Board

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Email: poper-at-nbacc.net
Name: Robert Pope

Organization: BNBI/NBACC

Title-Subject: [Filtered] Open Position: Research Assistant Electron Microscopy

Message: JOB TITLE: Research Assistant, Electron Microscopy
Company:Battelle National Biodefense Institute www.bnbi.org

PRIMARY FUNCTION

Provides biological sample preparation support for Scanning Electron Microscopy (SEM) and
Transmission Electron Microscopy (TEM) and performs Scanning and Transmission Microscopy of samples
to support the identification and characterization of biological agents and animal tissues.

MAJOR RESPONSIBILITIES:

Prepares samples containing biological agents for SEM and TEM analysis.
Operates and maintains NBACC Scanning Electron Microscope.
Operates and maintains NBACC Transmission Electron Microscope.
Operates and maintains NBACC Electron Microscopy ancillary equipment.
Supports the operation of the NBACC Core Electron Microscopy capability.
Works with a team of scientists collaborating to develop, manage, and analyze research and to
conduct bioforensic analyses critical to the mission of the NBACC.
Maintains records, analyzes technical and programmatic data and prepares analytical reports.
Presents results in written and oral formats.
Attends specialized training and reviews safety manuals and SOPS in order to maintain a safe
workplace environment.
Performs work with select agents in BSL2, BSL3 and BSL4 laboratories with appropriate training.
Identifies departures from the Quality Management System (QMS) and initiates actions to investigate
and prevent such occurrences.
Performs other assigned duties.

MINIMUM REQUIRED QUALIFICATIONS:
Bachelors degree (or equivalent) preferably in a biological science field. Experience in general
laboratory, biological, and electron microscopy techniques preferred.
Experience in specific techniques including embedding, fixing, sectioning, and staining of
biological samples for scanning and transmission electron microscopy desired.
Experience in the interpretation of SEM and TEM images for a variety of biological samples desirable.
General skills in light microscopy, phase contrast microscopy and fluorescent microscopy desirable.
Familiarity with imaging of BSL-2/3/4 viruses and bacteria, and familiarity with the relevant
safety, biosurety, decontamination, and quality assurance guidelines desirable.
Proficiency of oral and written communications essential.
Exemplary organizational skills with a proven track record of working effectively both independently
and as a team player.
Knowledge or experience with biocontainment facilities and procedures, laboratory safety, biosurety,
and decontamination desirable.
Knowledge or experience with Quality Assurance (QA) programs such as ISO 9001 or ISO 17025 and/or
GMP/GLP work environments desirable.
Must be a citizen of the United States.
Must be able to obtain an Interim secret clearance, leading to an eventual top secret clearance,
Suitability for Department of Homeland Security (DHS), a favorable adjudication of the Department of
Justice (DoJ) for select agents access, and enrollment in the Biological and Personnel Reliability
Program.
Current clearance(s) desirable.
Required to enroll in the Special Immunizations Program (SIP).

Apply at http://www.bnbi.org/careers.html

https://home.eease.com/recruit2/?id=3620041&t=1

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I have a client who needs to make a replica of a wet surface
(gelatin-like) for our field emission SEM. I've seen a reference to
polyvinylsiloxane or Xantropen Blue. Before I run around trying to locate
this stuff, does anyone have a recommendation for any material, plus a
source? Hints and tips?

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*
****************************************************************************

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Hi

Two (euro) cents more to Steve, Debby and Ken very good general comments :

As far I know, the Hitachi SU5500 is a HR SEM working in a TEM like
configuration, with the sample (cut to a smale size) in the OL, between
the upper and the lower pole piece, at WD = 0.
If the sample is a thin sample, and if the SEM is fited with a
transmited electrons detector (note the two "if"), it works in a way
similar to a STEM, and in that case the best resolution conditions will
be at high beam energy. Note that I say "resolution" conditions, not
"surface informations" conditions.
Look carefully at the indications on the picture or what is said in the
paper : which detector was used, SE (upper) or STEM ? What about the
sample preparation ? Is the picture a side or cross section like view,
or a top view ? Maybe the explanation of the choice of 30 keV is there.
Maybe too, that the conditions were choosen for STEM mode, but a picture
in SE-TTL was made, and gave THE information needed to answer THE question.
I'll hope that someone using a SU5500 know the way to use a SEM, but...

Hope it helps

--

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Mat�riaux de Strasbourg
D�partement Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail : Jacques.Faerber-at-ipcms.unistra.fr

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Email: crozier-at-asu.edu
Name: Peter Crozier

Organization: Arizona State University

Title-Subject: [Filtered] Want CCD camera for Gatan Image Filter

Message: Dear Colleagues,
The CCD camera on the back of our Gatan Imaging Filter (GIF) has failed and is apparently too old to
be repaired according to Gatan. The system is installed on an FEI Tecnai F20 TEM and it is the main
detector for our EELS capability. The electron optics for the GIF is fine it is just the detector
that has failed. We are interested in hearing from anyone that may have a suggestion on how to
repair the system. Ideally we would like to acquire a used CCD camera that is compatible with our GIF.
The model number of our current camera is 794GIF.3K3BP.33. It is a SI033 Site CCD. Any 794GIF in
the serial# range of 01102000 up to 04020200 will have this CCD.

We would also consider a complete replacement of the GIF with a used GIF or used PEELS system (we
dont perform image filtering on our Tecnai).

If any of you are able to help us secure this repair please contact me. We appreciate any
assistance and suggestions you may have.

Thanks

Peter Crozier
Associate Professor
Arizona State University

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***************************************************************************************
Forwarded from "Ask a Microscopist"
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****************************************************************************************
realname - Soya
Email - sgamsey-at-hotmail.com
EDUCATION - Graduate College
QUESTION - Is it possible to coat a porous sample (~80um pore size) in a
way that allows deposition of the coating material (e.g. Pt) into the pores?

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Hi

If you are considering sputter coating there is a method I have used but not
for pores as small as you have.

Place the sample in the sputter coater and pump it to attain the best
possible vacuum. Do not bleed gas into the system but try to force it to
coat; automated systems may not let you do this old systems will. The
theory is that with very little gas you have more straight line deposition,
the only chance of putting the coat inside the pores.

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Tel & Fax +44 (0)1280 816512 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
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Sent: 23 January 2013 17:03
To: protrain-at-emcourses.com

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Hi Soya,

Sputter and evaporative coating methods are line of sight to the
source. This means if you want to coat into a porous surface, you have
to play games with the geometry. The most common way would be with a
sample spinner, wherein the sample is doubly rotated on an eccentric
path, usually while tilted. This will only allow you to coat those
pores with line of sight to the source from any possible orientation.

-Jacob

On 13-01-23 09:12 AM, philip.oshel-at-cmich.edu wrote:
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}

--
Jacob Kabel
Electron Microscopist
Department of Materials Engineering
The University of British Columbia
6350 Stores Road, Room 419
Vancouver, British Columbia
Canada
V6T 1Z4
Phone: (604) 822-5648
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Dear All,
anybody out there who can send me the manual of the LKB Histo Knifemaker 2078 in PDF?

Thanks,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
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++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
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I'm going to defer to others for the specifics on this, but wouldn'
tan Osmium tetra oxide treatment work for getting into the porous
surface cavities? I know it's not exactly the most environmentally
friendly solution, but would that work better than the sputter coating
methods?

--Justin A. Kraft

On Wed, Jan 23, 2013 at 2:41 PM, {jkabel-at-mail.ubc.ca} wrote:
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} Hi Soya,
}
} Sputter and evaporative coating methods are line of sight to the
} source. This means if you want to coat into a porous surface, you have
} to play games with the geometry. The most common way would be with a
} sample spinner, wherein the sample is doubly rotated on an eccentric
} path, usually while tilted. This will only allow you to coat those
} pores with line of sight to the source from any possible orientation.
}
} -Jacob
}
} On 13-01-23 09:12 AM, philip.oshel-at-cmich.edu wrote:
} }
} }
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} } realname - Soya
} } Email - sgamsey-at-hotmail.com
} } EDUCATION - Graduate College
} } QUESTION - Is it possible to coat a porous sample (~80um pore size) in a
} } way that allows deposition of the coating material (e.g. Pt) into the pores?
} }
}
} --
} Jacob Kabel
} Electron Microscopist
} Department of Materials Engineering
} The University of British Columbia
} 6350 Stores Road, Room 419
} Vancouver, British Columbia
} Canada
} V6T 1Z4
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} http://www.mtrl.ubc.ca/research/eml/
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--
"America believes in education; the average professor earns more money
in a year than a professional athlete earns in a whole week." Evan
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Hi Soya,

Time of coat depends upon your coater, perhaps try 1 minute coats. Tilt 45
degs in one direction for the first coat, then 45 degs in the opposite
direction for the second. For the final coat move to the best vacuum as
described.

In my experience the higher vacuum coat is important because the object of
sputter coating is to cause the metal to be deviated away from line of site
by the gas. The better vacuum increases the mean free path of the metal and
that is the only way to penetrate holes.

An interesting experiment for the doubters is to take a nut (as in nuts and
bolts) about 1.16inch hole diameter and see which technique puts metal
through the hole.

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Tel & Fax +44 (0)1280 816512 Cell +44 (0)7711 606967
www.emcourses.com

-----Original Message-----
X-from: Soya Gamsey [mailto:sgamsey-at-hotmail.com]
Sent: 23 January 2013 18:39
To: protrain-at-emcourses.com

--

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Staff position in the EM Facility at New York Structural Biology Center

The New York Structural Biology Center (NYSBC) seeks an experienced
electron microscopist to join the staff of its cryo-EM facility. The
NYSBC is a shared center that supports state-of-the-art research in
cryo-EM, NMR, and X-ray crystallography. Cryo-EM facilities includes
four transmission electron microscopes, one dual-beam FIB/SEM, and
related support equipment for grid preparation, high pressure freezing,
and sectioning. In general, projects focus on 3D reconstruction of
biological assemblies, such as the atomic structure of membrane
proteins, subunit organization of macromolecular complexes, and cellular
anatomy of various tissues. The individual will carry out experiments in
support of collaborative projects with affiliated investigators,
including EM sample preparation, data collection, and image processing.
Day to day duties include setup and maintenance of electron microscopes,
training and assisting users from NYSBC's nine Member Institutions, and
record keeping. The individual should be capable of multitasking and
enjoy working with other people. Good communication skills are
essential. Qualified applicants should send a curriculum vitae and names
of three references to David Stokes (_stokes-at-nysbc.org
{mailto:stokes-at-nysbc.org} _). Salary and job title will be commensurate
with experience. A Ph. D. is desirable but not essential for this
position. The position is currently open and applications will be
reviewed continuously until the position is filled.

--
William J. Rice, Ph.D.
Senior Scientist, EM Facility Manager
New York Structural Biology Center
89 Convent Avenue, NY, NY 10027

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Soya,

This is a perfect sample for low vacuum or low voltage imaging if you have
those capabilities.

Also, you can try coating with carbon using a vacuum evaporator. In this
case the carbon is projected down on the sample with minimal direction
change. Thus chances are better to get some down into the pores. However,
do keep in mind that you need to evaporate in a good vacuum to get the
finest coating. This along with metal shadowing to more effectively cover
the surface of your sample may be adequate to minimize charge accumulation.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

On 1/23/13 4:58 PM, "protrain-at-emcourses.com" {protrain-at-emcourses.com}
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Dear colleagues,

I am planning a mouse study which has to be performed by a private company. In their protocol to prepare the mouse intestine for histopathological investigation, they describe their protocol like that:

"In brief, colons (one part of the longitudinally divided colons) are fixed as “Swiss-rolls” overnight in 10% neutral buffered formalin. Then colons are transferred directly into 70% ethanol, rolled, processed(Technicon Autotechnicon Mono Tissue Processor.This automated tissue processor sequentially submerses tissue specimens into solutions of fixative, dehydrant, clearant and paraffin wax at timed intervals)and embedded into paraffin blocks. Sections of the size of 5 µm will be cut for haematoxylin andeosin staining. "

As a cell biology this dehydration looks really minimalistic. Personally I would add more step but I suppose that the protocols used by pathologists for routine analysis have been adapted for higher throughput.
The purpose is to investigate modifications of the histology of the tissue (rough changes), no special staining here.

I would like to have the feedback of pathologists to know if this is usual to reduce the dehydration to one step of ethanol.
Many thanks in advance.

Stephane

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Good morning,
dear Stephane,

the description of process by the company’s information says:

“in brief“... and further “…sequentially submerses tissue specimens into solutions of fixative, dehydrant, clearant and paraffin wax at timed intervals)”.
Therefore this does not necessarily mean that there is only “one step of dehydrating agent = ethanol”!

So you and we do not know which protocol the private company for the automated tissue processor exactly will follow.
You will have to ask for the detailed, complete processing manual including all steps until paraffin blocking (see below) , but I am sure, you would be able also (if you are paying client) to “order” a processing schedule YOU would like to be implemented for “your” special tissue.

In routine pathological tissue processing – as I understand and as I know from our Histology Lab - a routine schedule (times depending on the material to be processed, here you will get the times for our so called “fat containing tissue”-processing schedule - ) will be
(further evaluation of resulting paraffin blocks –as you know- by means of EM/TEM usually not recommended but can be done by “re-processing” and “re-embedding” into resin):

- Fixation in 10% neutral buffered formalin = 4% buffered Formaldehyde solution (commercially available, usually PO4-buffered):
since most of tissues sent to our Histo-Pathology Lab have been immersed in fixative already for transport, after macroscopy/grossing the small tissue objects are put into the plastic specimen cassettes for subsequent processing into paraffin. Usually the thickness of a sample at least in one dimension does not exceed 5 mm, but most of such sliced specimens are in the range of 2 -4 mm thickness (occasionally for larger biopsies applying also pressure or vacuum using special “fixator” devices) up to overnight (5-10 hrs) with or without increased temperature up to 35°/40°C for special tissues application)

after grossing: storage of the tissue containing cassettes in fixative usually -at- RT until automated processing:

Depending on the processor type (modern ones are sucking solutions from a concentrated stock solution bottle and dilute to the concentrations you would like / want to use automatically) you have to program an appropriate processing schedule: e.g. duration of each step / iteration / temperature and concentrations.

So in our Histo-Lab there is the following schedule (for “fat-containing” tissues) with regard to the automated processor (a Sakira Tissue-Tek) used:

- at least 1/2 hr -at- RT or elevated T 4% buffered formaldehyde solution
(- a washing step in buffer would be desirable but usually is not done to save TAT)
- Transfer from fixative directly into (70% ) 76% Ethanol (or Isopropanol) (1-2x, each at least 1hr) 1 hr
- Ethanol 80% : 1 hr
- Ethanol 96%: 3 x 1 hr
- Ethanol 100%: 2 x 1 hr
- clearant(s): varying from lab to lab: in our Histolab specimens are transferred within the tissue processor from the 3rd Ethanol 100%-bath directly into xylene (3 x 1 hr each). NB: other Labs use Xylene substituents (like Histoclear, Limonene, or similar).
Paraffin wax: Paraffin (melting point 65°C)4 x 45 mins each.

After Paraffin-impregnation, tissues are blocked on the plastic cassette (“Ausgiessen”, cf. “modular embedding centers” consisting of paraffin dispensing unit, warming trays for cassettes and cooling plate), cooled and then are ready for sectioning.

Disclaimer: mentioning of names of companies or chemicals only for information. No financial interest, no affiliation.

Hope this is helpful for you,

Wolfgang

Wolfgang MUSS
EM-Lab
Pathology
SALK-LKH (Gen. Hospital)
SALZBURG, AUSTRIA

} -----Ursprüngliche Nachricht-----
} Von: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
} Gesendet: Donnerstag, 24. Jänner 2013 08:11
} An: Muß Wolfgang
} Betreff: [Microscopy] dehydration for histopathology in mouse
}
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}
} Dear colleagues,
}
} I am planning a mouse study which has to be performed by a private
} company. In their protocol to prepare the mouse intestine for
} histopathological investigation, they describe their protocol like
} that:
}
} "In brief, colons (one part of the longitudinally divided colons) are
} fixed as “Swiss-rolls” overnight in 10% neutral buffered formalin. Then
} colons are transferred directly into 70% ethanol, rolled, processed
} (Technicon Autotechnicon Mono Tissue Processor. This automated tissue
} processor sequentially submerses tissue specimens into solutions of
} fixative, dehydrant, clearant and paraffin wax at timed intervals) and
} embedded into paraffin blocks. Sections of the size of 5 µm will be cut
} for haematoxylin andeosin staining. "
}
} As a cell biology this dehydration looks really minimalistic.
} Personally I would add more step but I suppose that the protocols used
} by pathologists for routine analysis have been adapted for higher
} throughput.
} The purpose is to investigate modifications of the histology of the
} tissue (rough changes), no special staining here.
}
} I would like to have the feedback of pathologists to know if this is
} usual to reduce the dehydration to one step of ethanol.
} Many thanks in advance.
}
} Stephane
}
}
} ==============================Original
} Headers==============================
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} 4, 30 -- From: Stephane Nizet {nizets2-at-yahoo.com}
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20, 39 -- Subject: [Microscopy] Re: dehydration for histopathology in mouse
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Dear Stephane,

I'm not familiar with the Autotechnicon Mono, but from Googling, it seems
that the tissue processor does multiple stages of dehydration.

I'm not sure why there is the preceding step of directly placing into 70%
ethanol� just guessing, but maybe if this is left out the rolls of tissue
are not tightly/compactly enough rolled after the automated dehydration?

Regards,

Ben
--
Research Support Manager
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271649
{http://www.mrc.ox.ac.uk/}

On 24/01/2013 07:14, "nizets2-at-yahoo.com" {nizets2-at-yahoo.com} wrote:

}
} Dear colleagues,
}
} I am planning a mouse study which has to be performed by a private
} company. In their protocol to prepare the mouse intestine for
} histopathological investigation, they describe their protocol like that:
}
} "In brief, colons (one part of the longitudinally divided colons) are
} fixed as �Swiss-rolls� overnight in 10% neutral buffered formalin. Then
} colons are transferred directly into 70% ethanol, rolled,
} processed(Technicon Autotechnicon Mono Tissue Processor.This automated
} tissue processor sequentially submerses tissue specimens into solutions
} of fixative, dehydrant, clearant and paraffin wax at timed intervals)and
} embedded into paraffin blocks. Sections of the size of 5 �m will be cut
} for haematoxylin andeosin staining. "
}
} As a cell biology this dehydration looks really minimalistic. Personally
} I would add more step but I suppose that the protocols used by
} pathologists for routine analysis have been adapted for higher throughput.
} The purpose is to investigate modifications of the histology of the
} tissue (rough changes), no special staining here.
}
} I would like to have the feedback of pathologists to know if this is
} usual to reduce the dehydration to one step of ethanol.
} Many thanks in advance.
}
} Stephane

==============================Original Headers==============================
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14, 31 -- Subject: Re: [Microscopy] dehydration for histopathology in mouse
14, 31 -- Thread-Topic: [Microscopy] dehydration for histopathology in mouse
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Email: yamawaki-at-stanford.edu
Name: Ruth Yamawaki

Organization: Stanford University

Title-Subject: [Filtered] Digital camera for JEOL CX100

Message: We are finally entering the digital age and are looking for a digital camera for our JEOL
CX100.

Please send me any tips/comments/service on your digital camera that can help us make an informed
decision.

Thank you,
Ruth Yamawaki
Stanford University
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650 723-3457

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Email: z.zhou-at-lboro.ac.uk
Name: Zhaoxia Zhou

Organization: Loughborough University

Title-Subject: [Filtered] Keep FEGTEM column clean

Message: We have a FEI Tecnai F20 TEM. Just installed, column nice and clean. STEM/HAADF mode hours
not seen the scan box/lines at all. Minimal contamination.

However, Ive got requests to look at some sol-gel prepared nanoparticles. Would heating the samples
(nanoparticles on holey carbon film) in an oven ~150°C for an hour be an effective way to prevent
possible contamination to the TEM? Are there other methods colleagues would like to recommend? What
are the usual procedures to keep an FEGTEM column clean with a wide range of types of samples?

Kind regards,
Zhaoxia Zhou

Dr Z Zhou
Experimental Officer
Loughborough Materials Characterisation Centre (LMCC)
Department of Materials
Loughborough University
Leicestershire
LE11 3TU
UK

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The University of Texas at Dallas has an opening for a postdoc position on the TEM/STEM study on nanostructured materials.

A Ph. D. in Materials Science or related physical science with experience on Transmission Electron Microscopy; knowledge of nanomaterials materials (especially, graphene, BN, MoS2, multilayer GaN/InN, etc.) and their structure-property relationships. Extensive experience with advanced electron microscopy documented by high-quality publications; hands-on experience on HRTEM and STEM with analytical techniques (EELS and EDS).

Interested candidates should provide a cover letter, CV, and a list of references to jinguo.wang-at-utdallas.edu. Applications will be evaluated as received.

Thanks

Jinguo Wang, Ph.D
Mail Station RL10
Department of Materials Science
University of Texas at Dallas
800 W. Campbell Road
Richardson, TX 75080-3021

jinguo.wang-at-utdallas.edu
972-883-6928

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Zhaoxia,

XEI Scientific Inc have a "TEM Wand" which replaces the sample-probe and
cleans up the sample region in a TEM by injecting Oxygen radicals into the
pole-region space.

Best Regards

Ian

Ian Holton
Acutance Scientific

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Email: z.zhou-at-lboro.ac.uk
Name: Zhaoxia Zhou

Organization: Loughborough University

Title-Subject: [Filtered] Keep FEGTEM column clean

Message: We have a FEI Tecnai F20 TEM. Just installed, column nice and
clean. STEM/HAADF mode hours not seen the scan box/lines at all. Minimal
contamination.

However, Ive got requests to look at some sol-gel prepared nanoparticles.
Would heating the samples (nanoparticles on holey carbon film) in an oven
~150°C for an hour be an effective way to prevent possible contamination to
the TEM? Are there other methods colleagues would like to recommend? What
are the usual procedures to keep an FEGTEM column clean with a wide range of
types of samples?

Kind regards,
Zhaoxia Zhou

Dr Z Zhou
Experimental Officer
Loughborough Materials Characterisation Centre (LMCC) Department of
Materials Loughborough University Leicestershire
LE11 3TU
UK

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Dear Collective,

Somewhere in the dim mists of memory seems to lurk a notion that I once saw a protocol for growing bacteria directly on carbon-coated TEM grids. So far, I've been unable to locate any trace of it, so either I have lost it or I am losing it. Does anybody know of techniques for doing this?

Thanks for any help you can give me.

Cheers,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

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Me, again. I forgot the second part of my question about culturing bacteria---are there ways of removing excess extracellular polysaccharides from bacterial cultures in order to cut down on the background garbage in negative stain? Our client is going to try a "protocol gradient", if I understand him correctly (I please ignorance on this), but we're coming up short on other ideas.

Thanks again!

Cheers,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

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Long, long ago, when using the Periodic Acid Schiff (PAS) Rx on 'fresh', paraffin-embedded (that means well cooked, dewaxed, dehydrated and fixed (factually denatured)) tissue sections (all of that done to something like liver so it would look like the picture Maximov drew with his colored inks and published in one of his many histology HARDBOUND books on histology*) those of us who required a 'control' section were EXPECTED to acquire it by processing a second section with freshly collected saliva either collected solely by the efforts of one or, of course, better statistically if acquired thru contributions of a large, representative group of saliva producers. The result was more magenta in the test and less in the control. Pure science!

Using that method, one NEVER considered a gradient of anything in a protocol. However, one could, even in that dark age, purchase a pure example of an appropriate enzyme from Worthington.

* Histology has almost disappeared as a subject that synthesizes, anatomy, physiology, development, and often some pathology. My guess is that it requires sufficient knowledge of anatomy, microscopy, and those other subjects to build a curriculum of obligatory study - something that does not ring the school bells these days. Except for one exception, written for pathologists, normal histology now comes with a soft cover just to prove that it is not going to last much longer.

Cheers,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

HomePage: http://cmirt.wcupa.edu (with link to instrument Scheduler)

-----Original Message-----
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Me, again. I forgot the second part of my question about culturing bacteria---are there ways of removing excess extracellular polysaccharides from bacterial cultures in order to cut down on the background garbage in negative stain? Our client is going to try a "protocol gradient", if I understand him correctly (I please ignorance on this), but we're coming up short on other ideas.

Thanks again!

Cheers,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

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Dear Randy,

since I have not seen any reaction or reply to your post (1) with
the exception of Fred Monson's post on TEM Neg staining(2)

I would like to point you cautiously to the following papers

---------------------------------------------------------------------
cf.
www.bss.phy.cam.ac.uk/~es10009/documents/thomson_scanning2011.pdf

= SCANNING VOL. 33, 59-68 (2011)
& Wiley Periodicals, Inc.
=====Imaging Internal Features of Whole, Unfixed Bacteria=======
NICHOLAS M. THOMSON1, KEVIN CHANNON1, NOOR AZLIN MOKHTAR2,
LECH STANIEWICZ1, RANJANA RAI3,IPSITA ROY3, SHUN SATO4,
TAKEHARU TSUGE4, ATHENE M. DONALD1, DAVID SUMMERS2, AND
EASAN SIVANIAH1
1Cavendish Laboratory, University of Cambridge, Cambridge, UK
2Department of Genetics, University of Cambridge, Cambridge, UK
3Department of Molecular and Applied Biology, University of Westminster,
London, UK
4Department of Innovative and Engineered Materials, Tokyo Institute of
Technology, Yokohama, Japan
DOI 10.1002/sca.20221 Published online 22 February 2011 in
Wiley Online Library (wileyonlinelibrary.com)
--------------------------------------------------------------------
Most important with regard to your initial quest:

==== In situ multiple sampling of attached bacteria for scanning
and transmission electron microscopy.=====
Bozzola JJ, Johnson MC, Shechmeister IL.
Stain Technol. 1973 Nov;48(6):317-25. No abstract available.
PMID: 4129036 [PubMed - indexed for MEDLINE]
(If you need a / this .pdf, please just request it with your reply)

--------------------------------------------------------------------
and cf. (free access) -at-
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC246670/pdf/jbacter00340-0326.pdf
J Bacteriol. 1974 April; 118(1): 304-311. PMCID: PMC246670
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC246670/
===== Morphological Study of Streptococcus mutans and
Two Extracellular Polysaccharide Mutants =========

by M. C. Johnson, J. J. Bozzola, and I. L. Shechmeister

ex methods (it is more/less the same method as used in the
Stain Technol. 1973 Nov;48(6):317-25-article):

Electron microscopy.
A technique was developed which allowed electron micrographs to be taken of bacteria as they appeared when grown on a hard surface in a liquid medium containing sucrose. In this procedure,
a sterilized microscope slide,
a cover slip, and

{ {a cover slip with Formvar-carbon-coated grids were placed in a sterile plastic petri dish} } .

Sucrose broth was added, inoculated with the desired organism, and incubated at 35 C for a predetermined time. The above components were then treated as follows. The slide and plain cover slip were removed and gently washed two to three times in double-distilled water; the slide was air-dried for three to four days, and the cover slip was fixed overight in 3% gluteraldehyde and buffered to pH 6.0 with Veronal acetate.

The dried slide was processed for replicas (17), and the

cover slip was rinsed thoroughly in double-distilled water, air- or freeze-dried, coated with 40 to 50 nm of palladium-gold (60 to 40 alloy), and examined in a Kent Cambridge Mark II Stereoscan operating at an accelerating voltage of 30 kV and at a 450 angle tilt.

{ {The cover slip with attached grids} }
was dipped three times in double-distilled water, and the grids were removed to a filter paper and stained with 2% phosphotungstic acid at pH 5.5.

The bacteria on the bottom of the petri dish were then embedded in situ as follows: after a gentle wash with Veronal acetate, the sample was fixed for 24 h in Veronal acetate-buffered 1% OSO4 (16), dehydrated, and gradually infiltrated with Epon at 0.5 h-intervals by three changes of 3:1, 1:1, 1:3 (vol/vol) mixtures of absolute ethanol-Epon. After three more changes of pure Epon, a thin layer of resin was poured onto the petri dish and polymerized at 60 C for 48 h. The hardened plastic was allowed to cool for 24 h, after which the Epon layer was stripped off, sectioned, and examined with a Hitachi HU-11A transmission electron microscope at an accelerating voltage of 50 kV.

---------------------------------------------------------------------------
and last but not least (old too, but perhaps also interesting, and
Reinhard Rachel, Regensburg,
who is also on the MSA-List can/will comment on that):

J Bacteriol. 2006 October; 188(19): 6915-6923.
doi: 10.1128/JB.00527-06
PMCID: PMC1595509
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1595509/
Flagella of Pyrococcus furiosus: Multifunctional Organelles,
Made for Swimming, Adhesion to Various Surfaces, and Cell-Cell Contacts
Daniela J. N�ther,1 Reinhard Rachel,1 Gerhard Wanner,2 and Reinhard Wirth1
Adherence studies: growth on carbon-coated gold grids for TEM.
Methods to study growth of microorganisms
{ { directly on carbon-coated gold grids used for TEM } } have been
developed in our labs ([39] notice: + Ref.: see below).
Gold grids were placed in small Teflon holders in serum bottles
containing anaerobic medium. For TEM cells were fixed with 2.5%
(final concentration) glutaraldehyde for 30 min at room temperature.
In the case of cell or flagellum suspensions, a drop was placed on
a carbon-coated 200-mesh copper grid (Plano, Wetzlar, Germany).
The samples were either unidirectionally shadowed with Pt/C at
15� (CFE 50; Cressington Ltd., Watford, United Kingdom) or
negatively stained for 1 min with 2% uranyl acetate.
All TEM micrographs were recorded using a slow-scan charge-coupled
device camera (TEM 1000; TVIPS-Tietz, Gauting, Germany) attached
to a CM 12 transmission electron microscope (FEI, Eindhoven,
The Netherlands) operated at 120 keV.

[39]. Rieger, G., R. Rachel, R. Herrmann, and K. O. Stetter. 1995.
Ultrastructure of the hyperthermophilic archaeon Pyrodictium abyssi.
J. Struct. Biol. 115:78-87.
unfortunately this article does not show up in PubMed and
did not result as a Google-search finding.
------------------------------------------------------------------------

There also one could consider to use "lacey Support-Film" Grids:

http://www.tedpella.com/supflm_html/suptfilm.htm
7. Lacey Support Films - NetMeshT Grids:

Lacey Support Film

A lacey network support film. The holes in the lacey support film
vary in size from less than a quarter micron to more than 10 microns
making them ideal for any type of specimen. Lacey support films are
extremely strong and withstand vigorous specimen preparation treatment.

The specimen material is supported by the film network but lies across
or protrudes into the holes of the mesh. This allows high definition
imaging without the effects of underlying support material. Lacey films
can be used for specimens ranging from large crystals and other
particulate material to virus particles. Smaller particles,
such as viruses or bacteria,
tend to adhere around the inner edges of the holes, an ideal situation
for high resolution microscopy. Lacey films are also ideal for selected
area electron diffraction imaging. We offer three types of lacey film:

http://www.tedpella.com/supflm_html/suptfilm.htm
Support Film Grids,

Substrate Application Guide
B= Best; G= Good Alternative; -= Not Suitable
------------------------------------------------------------------------x
Substrate Formvar Formvar Silicon Silicon Carbon Carbon
Application Only Stabilized Monoxide Monoxide Type-A Type-B
with Carbon on Formvar on Type-A

Bacterial -NO- G B B B B
Suspensions
------------------------------------------------------------------------x

x-----------------------------------------------------------------------z
Substrate Pure Is Lacey Film
Application Carbon suitable for
Film this application?
Bacterial
Suspensions B Yes
x-----------------------------------------------------------------------z

Hope this helps you at least a little bit to get started,
sorry again (apologizing) for the longness of my posting
best regards and
to all of you listers
a happy and joyful weekend,

Wolfgang MUSS PhD
Salzburg - Austria

} -----Urspr�ngliche Nachricht-----
} Von: tindallr-at-missouri.edu [mailto:tindallr-at-missouri.edu]
} Gesendet: Donnerstag, 24. J�nner 2013 18:03
} An: Mu� Wolfgang
} Betreff: [Microscopy] TEM Neg staining(1): bacteria grown on carbon
} films
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} Dear Collective,
}
} Somewhere in the dim mists of memory seems to lurk a notion that I once
} saw a protocol for growing bacteria directly on carbon-coated TEM
} grids.
}
} So far, I've been unable to locate any trace of it, so either I have
} lost it or I am losing it.
} Does anybody know of techniques for doing this?
}
} Thanks for any help you can give me.
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior Research Specialist
} Electron Microscopy Core Facility
} W125 Veterinary Medicine
} University of Missouri
} Columbia, MO 65201
} (573) 884-5383
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Dear Judy,
unfortunately I have not seen any response to your post via MSA-Listserver. You might have gotten some reply's on this, so apologize if I am bothering with my posting.
The procedure of collecting blood cells for TEM most frequently is the preparation of a so called "Buffy Coat" (in the literature or in www- recipes I have found several misspellings for this term).

So, just to get started (there may exist hundreds of other or more sophisticated protocols):

1) Usually one will collect platelets from peripheral blood (the more ml the better - knowing this to be an ethical question)
2) Use e. g. EDTA-peripheral blood (filled into 'Monovettes' or similar pre-treated tubes with EDTA for blood sampling). There are out also Heparin-layered tubes. Avoid/ DON'T use collecting systems containing small glass globules! You'll never get a {buffy coat} .
2) You'll need a preparatory centrifuge, for preparing a "buffy coat" a centrifuge with a swing-out rotor is suited best (there are some small centrifuges which have implemented not only fix angle rotor, but swing out rotor as well, if you own a (cooled) bench top centrifuge you for sure are on the right side)
NB: cf. e.g. http://www.hettichlab.com/appc/content_manager/page.php?ID=160391&dbc=fa06a43d4d0ce718387d80170d8decd7
See: e.g. Small centrifuges: EBA 270, EBA 21; or BenchTop centrifuges...+/- all types, spec. rotors are parts to order)
(Disclaimer: there are a lot of centrifuge manufacturers. This URL only for a rough information. No financial interest, no affiliation, just having been satisfied user of one of their products over many years)
3) Before centrifugation topple tubes carefully / with caution some times (3-4 times)
4) make sure you'll have at least 2 tubes with the same weight (total tube+blood)
5) centrifuge blood for 10 min -at- 1000 g (the g-force will depend on radius of the rotor as well as the velocity, so you'll have to check this with the centrifuge's nomogramm which normally is enclosed with the centrifuge manual)
6) don't use a "break-function" of the centrifuge, that means, after centrifugation let swing out the rotor without disturbing the slowing of rotation until the rotor stands still

7) Carefully/cautiously take out the vials - put them into a fitting (tube)rack.
8) You have prepared in advance your fixative for TEM (there are a lot of possibilities, as you know), use at RT
9) In the Monovettes/tubes you should have gotten a "Buffy coat", that means layers of different dense "fluids" (from above to bottom), containing:
- Plasma/Serum (somehow yellowish-brown, depending on lipids in serum
- a thin layer of somehow whitish substrate (= platelet fraction)
- a thin layer of red/reddish substrate ( on top of that layer you'll find lymphocytes, on ground of that layer you 'll have spinned down neutrophils etc.
- bottom (+/less thick): RBC-Red blood Cells.
CAVE: don't use rotation forces higher than maximally recommended for the plastic tubes! 1.000 g is enough!

Take care when handling this because it is all fluid and the layer formation can be disturbed by any breath of a 'kick'.

Fixation of "Buffy Coat":
10) Gently/carefully open the tubes, suck off Plasma by means of a long pipette tip (slowly and positioning the tip in the middle of the tube. Control removal of serum carefully. Leave only a small layer of serum over the first opaque looking layer (taking care not to suck off from that layer with the pipette).
11) Gently release your TEM- fixative solution ONLY DROP BY DROP from another pipette tip which you place closely to the inner wall of the tube. Observe initial mixing of plasma remnants with the fixative. NO WHIRLING should be produced with that initial step of fixation.
(Cave: the thicker the left plasma layer will be the less you'll face problems with whirling, on the other hand you'll need longer time to get initially fixed the cells below (i.e. platelets, lymphocytes, neutrophils, etc).
After some minute add more drops of fixative slowly (at the end, fill up the tube)
If you use initially a primary fixative (e. g. 0.5 % FA + 1.5% GA buffered n 0.1M PO4-(or sodium cacodylate) buffer, let stand at RT for at least 15 -30 min UNDISTURBED on its own. Afterwards you can suck off the primary fixative and fill in cautiously your main fixative (e.g. 3-4% GA buffered in 0.13M PO4 = isotonic for blood or 0.1M cacodylate buffer).
12: let stand for either at least 3 hrs at RT or try out also "mild" MW-fixation in the tubes.

Collection of "Buffy Coat":
Depending on the type of tubes you have used (plastic ones are favorite, but also with glass tubes there is a possibility of breaking the glass by means of a glass cutting device and special pliers, which could be produced by a tool shop) you have to get rid of the bottom part of the tube. Remember that the bottom part (RBC's) will not have been fixed properly at that time (so use PSA, at least gloves!).
Cut the bottom part some mm below the RBC-layer, place tube into fixative again (swirling a little bit to get rid of loose and unfixed RBC's). then try to separate the whole layer-plug from the tube wall (there are some possibilities how to manage this) and try to push out the whole fixed plug.
Dissect under stereomicroscope: you certainly will see the different layers, among them the layer of thrombocytes / platelets right below the small fixed plasma layer.

Another possibility to collect platelets perhaps would be density gradient centrifugation (but this - with e.g. the SIGMA Histopaque�-Ficoll method usually is done to remove all platelets from the MNC-fraction).
Please also consider other techniques depending on the purpose of your intended studies.
If you have any question left, let me know
(also off list if you prefer... I have done a lot of those {buffy coats} in my "career"),
Best wishes and regards
Wolfgang

Wolfgang MUSS
EM-Lab
Pathology,
SALK-LKH (Gen. Hospital) SALZBURG
AUSTRIA

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} An: Mu� Wolfgang
} Betreff: [Microscopy] Prep for platelets for TEM
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} Title-Subject: Prep for platelets for TEM
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Dear Listers;

I have a faculty member who wants to embed brain tissue pieces that are 100-200 microns thick, 2cm in length and 1 to 2cm in width in LR White. I have Aclar film and have located PTFE flat molds with large enough cavities that may work. Before I proceed I would like to hear from anybody who may have experience in doing this kind of embedding. Any advice is appreciated. I am especially interested in knowing if the Aclar film will really work well enough to seal the mold and prevent air from interfering with the polymerization. Also would it be best to first use UV polymerization in a cold chamber or can I go directly to the embedding oven at 65 degrees C? Could variable wattage microwave polymerization be used without submerging the mold underwater? As always thanks in advance for the help.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

==============================Original Headers==============================
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Tom
Do you have access to an oven with vacuum? If so then just connect to a nitrogen source and alternate pulling a vacuum and the flooding with nitrogen to drive out the oxygen. Then leave the oven at 60oC to polymerize. Only leave vacuum low- at about 5lb - so as not to force resin to creep up the molds.
Debby

DS imaging LLC
Www.dsimagingllc.com

Sent from my iPhone

On Jan 29, 2013, at 3:55 PM, "tbargar-at-unmc.edu" {tbargar-at-unmc.edu} wrote:

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} Dear Listers;
}
} I have a faculty member who wants to embed brain tissue pieces that are 100-200 microns thick, 2cm in length and 1 to 2cm in width in LR White. I have Aclar film and have located PTFE flat molds with large enough cavities that may work. Before I proceed I would like to hear from anybody who may have experience in doing this kind of embedding. Any advice is appreciated. I am especially interested in knowing if the Aclar film will really work well enough to seal the mold and prevent air from interfering with the polymerization. Also would it be best to first use UV polymerization in a cold chamber or can I go directly to the embedding oven at 65 degrees C? Could variable wattage microwave polymerization be used without submerging the mold underwater? As always thanks in advance for the help.
}
} Tom Bargar
} UNMC
} Electron Microscopy Core Facility
} 986395 Nebraska Medical Center
} Omaha, NE 68198-6395
} 402-559-7347
} tbargar-at-unmc.edu
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6, 30 -- From dsherman-at-purdue.edu Tue Jan 29 15:03:20 2013
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Hello Tom,

Yes, LR White can be tricky, but we have a method that seems odd but always works. In our experience it is better to not fill all the PTFE mold spaces with samples. We have to orientate drosophila retina and larvae very strategically, so we place our samples into the center 6 wells, overfill the wells including two extra empty ones on either side of the samples. Press the Aclar film over the samples in the middle first then gently lay out the film toward both sides so that the resin pours over into the left-over empty well spaces toward the ends. The very end well spaces will not polymerise very well, but the ones with the samples will turn out very well, at least for us. We then place the mold into a 60 degree oven overnight to two days, sometimes three depending on the weather. So far we have been able to thick section and do fluorescence staining with no problem. Hope this helps.

Lita Duraine
EM Technologist
Bellen Lab
HHMI- Molecular Genetics
Duncan Neurological Research Institute
1250 Moursund St.
Houston, TX 77030
Rm: N1165.17
MS: N1125.50
832-824-8772 TEM Room
979-549-6526 Cell
http://flypush.imgen.bcm.tmc.edu/lab/people/lita.php

-----Original Message-----
X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu]
Sent: Tuesday, January 29, 2013 3:05 PM
To: Duraine, Lita

Dear Listers;

I have a faculty member who wants to embed brain tissue pieces that are 100-200 microns thick, 2cm in length and 1 to 2cm in width in LR White. I have Aclar film and have located PTFE flat molds with large enough cavities that may work. Before I proceed I would like to hear from anybody who may have experience in doing this kind of embedding. Any advice is appreciated. I am especially interested in knowing if the Aclar film will really work well enough to seal the mold and prevent air from interfering with the polymerization. Also would it be best to first use UV polymerization in a cold chamber or can I go directly to the embedding oven at 65 degrees C? Could variable wattage microwave polymerization be used without submerging the mold underwater? As always thanks in advance for the help.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

==============================Original Headers==============================
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One of the outstanding characteristics of turbomolecular pumps is
that they develop such high compression ratios for oil molecules that
these molecules are unable to move from the backing line, through the
pump, and into the vacuum system - provided the pump's rotor is
turning at nearly full rotational speed. This, combined with the fact
that they can produce pressures in the UHV range, makes them
particularly attractive as pumps on electron microscopes. HOWEVER, if
the turbo pump is backed by an oil-sealed rotary vane pump achieving
this contamination-free operation requires very careful adherence to
proper pump-down and venting procedures. This important matter is
discussed in considerable detail in Section 6.1.9b (pages 255 - 258)
of my book, "Vacuum Methods in Electron Microscopy".

Now-a-days, however, a more straightforward solution is to use an
oil-free backing pump, such as a scroll pump. If you replace a rotary
vane pump with a scroll pump it would be wise to also replace the
backing line with a new oil-free line.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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Email: trent-at-ornl.gov
Name: shawn reeves

Organization: Oak Ridge National Lab

Title-Subject: [Filtered] RuO4 staining

Message: Hi,

I will be setting up an area to stain polypropylene blocks with RuO4. I'll be using the kit sold
from SPI which consist of RuO2 hydrate and sodium periodate. I'm wondering if anyone could guide me
on the safety of this process and how to dispose of the product. i think this kit produces 0.67%
aqueous RuO4 and I will need the solution to be 0.5% to dispose of it. How should I treat
everything that has come in contact with the solution, vials, tissues etc. Also I will be slicing
the polypropylene with our microtome, is there any safety issues concerning the stained
polypropylene which will be falling in the microtome?

Thank you everyone in advance for your replies!

shawn

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Email: margaret.dienelt-at-ars.usda.gov
Name: Margaret Dienelt

Organization: Agricultural Research Service, USDA

Title-Subject: [Filtered] Need manual for EM Tech K850 Critical point dryer

Message: Hello Everyone,

We have an old EM Tech K850 critical point dryer in our lab that has never been used. After all
these years, we need to use it but of course the manual is nowhere to be found. Does anyone have a
manual to this instrument they can spare? If you have a manual but can't spare it, would you please
share the contact information of EM Tech with us?

Another option: if you can spare your manual for a week, we will copy it at our expense and send it
back the next day.

Thank you very much.

Margaret Dienelt
Plant Pathologist/Electron Microscopist
Floral and Nursery Plants Research Unit
NA/ARS/USDA

Bldg 012 Rm 1-7
10300 Baltimore Avenue
Beltsville, MD 20705 USA

Phone: 760-263-4219, 301-504-6097
Email: margaret.dienelt-at-ars.usda.gov
Fax: 301-504-5096

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Email: joseph.uknalis-at-ars.usda.gov
Name: Joe Uknalis

Organization: USDA-ERRC

Title-Subject: [Filtered] Infiltrate EMbed 812 into gram positive bacteria

Message: I've had difficulty getting good infiltration into Listeria and Staphlococcus. I've tried
long infiltration times (2 days) and under house vacuum.

Is the secret to good infiltration even longer times, higher vacuum levels or ???

thanks

Joe

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Greetings Fellow Microscopists,

Hooke College of Applied Sciences, located in Westmont, IL, is offering a scanning electron microscopy short course March 4-8, 2013. In addition to lectures, this course emphasizes hands-on training using state-of-the-art equipment. For further SEM training details and registration information, please follow the link below:

http://www.hookecollege.com/courses/course.asp?COURSE_ID=42

Best regards,

Chris Gorman
Hooke College of Applied Sciences
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7412 (fax)

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Dear Colleagues,

We would like to bring to your attention three symposia at the
upcoming M&M 2013 meeting,
to be held in Indianapolis, Indiana on August 4-8, 2013. We would like
to encourage you
to submit a short paper and join us in Indianapolis.

This year the M&M conference will begin with a plenary session
featuring a talk by
Prof. Harald Rose, “The Long-Lasting Struggle to Achieve
Atomic-Resolution Microscopy
by Correcting the Aberrations of Electron Lenses". In his presentation
Prof. Rose will
provide an overview of the history of electron microscopy, and give
key insights into
the challenges of electron microscopy in the future. The second
keynote speaker will
be Joris Dik with a talk entitled: "Looking Through Paintings" With a
background in
Art History, Chemistry and Materials Sciences, Professor Dik brings a
unique perspective
to the study of paintings and masterworks, combining insights from
both the science
and the art worlds.

The three symposia related to structure, ultrastructure, and
technology development are:
_________________________________________________________________________________________
A04 Electron Tomography in Life and Material Science
Organizers: Heiner Friedrich, Montserrat Barcena, Esther Bullitt

Advances in sample preparation, instrumentation and methodology have
widened the scope
of electron tomography (ET) from sub-nanometer details to the
micrometer scale. This
symposium will address leading scientific and technological
developments in the physical
and biological sciences, using the widest possible range of ET imaging
approaches and
their integration with complementary techniques. Applications and
developments covering
— but not limited to — (aberration-corrected, energy-filtered) TEM
and STEM, phase plates,
diffraction or holography, are invited. Contributions including
complementary approaches
such as correlative light-electron microscopy, X-ray tomography,
serial-block face
SEM/AFM, and novel processing tools are encouraged.

Invited presenters: David Mastronarde, Elizabeth Wright, Irene Wacker,
Dirk van Dyck,
Krijn de Jong, Daniel Wolf
____________________________________________________________________________________________
A09 Advances in Data Processing in Optical and Electron Microscopy
Organizers: Edward P. Morris, David Morgan, Jeffrey L. Clendenon

Many advances in microscopy have been driven by developments in
software and the availability
of affordable high-speed computing. This symposium will focus on
software tools for EM and
LM that are publically available with an emphasis on analysis after
data acquisition. A major
topic will be EM software for structural biology (analysis of single
particles, helical
structures, 2d crystals), TEM/STEM tomography and enhancing
information obtained from
materials. Another focus will be software developed for LM to process,
visualize, segment
and measure 3D/4D images.

Invited presenters: Kevin Eliceiri, James Glazier, Michael
Radermacher, Hanspeter Winkler,
Quentin Ramasse, Lewys Jones

____________________________________________________________________________________________
B03 Structural Biology and Cell Ultrastructure
Organizers: Paula C. A. da Fonseca, Michael Radermacher, Ingeborg Schmidt-Krey

Our understanding of the 3D structure and function of cells,
microorganisms and macromolecular
assemblies has experienced great advances through recent developments
of EM techniques and
hybrid methodologies. This symposium highlights structural and
ultrastructural studies of cells,
microorganisms and biological macromolecules using electron microscopy
techniques
(e.g. single-particle analysis, tomographic methods; helical
reconstruction, crystallographic
methods) singly or combined with other structural methods (e.g. X-ray
methods; atomic force
microscopy). Topics will include: structure and function of
macromolecular assemblies, virus
structure and virus-host interactions; eukaryotic and prokaryotic cell
architecture; cellular
metabolism; cell division and protein translation; cellular secretion,
adhesion and motility;
cell-cell communication and signaling.

Invited presenters: David Veesler, Edward Morris, Beate Rockel, Yifan
Cheng, Tamir Gonen,
Timothy Baker
_____________________________________________________________________________________________
Please also spend some time to browse the complete program and you
will discover many other
presentation of high interest to our field.

Specifics:
The abstract/short paper submission deadline is on February 15th. The
submission website will
close at 5PM PST on this date. Contributions must be 2-pages long and
have at least 300 words.
Note that 1-page or shorter contributions might be rejected and, if
accepted for presentation,
will not be part of the proceedings. The 2-page meeting contributions
can be deposited under
the public access policy of granting agencies (e.g. Pubmedcentral / NIH).

When you submit your paper, select any of the three symposia:
A04 Electron Tomography in Life and Material Science
A09 Advances in Data Processing in Optical and Electron Microscopy
B03 Structural Biology and Cell Ultrastructure

Please see the full paper submission guidelines on the meeting website.
Link to meeting site: http://www.microscopy.org/MandM/2013/callforpapers.cfm
This is a great educational and networking meeting for the entire
microscopy community.
Students can benefit from a reduced registration fee ($50.00) and have
ample opportunities
to receive awards either before the meeting (make sure to check the
corresponding box during
submission) or during the poster session. For more information go to:
http://www.microscopy.org/MandM/2013/callforpapers.cfm

When registering, please also check out the many interesting Sunday
pre-meeting courses
and workshops.

We look forward to seeing many of you at the meeting.

Sincerely,
The organizers of A04, A09, and B04

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Hi all,
I would like your advice/opinion on critical point dryers and sputter-
coaters. We are leaning towards the Tousimis 931 cpd (winner of the
2012 Microscopy Today Innovation Award) and the SPI Module sputter/
carbon-coaters.
thanks in advance,
Beth

==============================Original Headers==============================
2, 20 -- From beth-at-plantbio.uga.edu Wed Jan 30 15:34:52 2013
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Hi Beth,
We recently purchased the Tousimis 931. I absolutely love it!!!!!!! It's
awesome. The samples I processed have been superb!!! It has tremendously
freed up my time and has been completely dependable.
I'm not familiar with the SPI sputter/carbon coaters. We have a PELCO SC-7
Sputter coater and it's probably the best one I've ever used. It's easy and
quick. However it is just a low vacuum sputter coater(0.08mbar).
Hope that helps!
Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

-----Original Message-----
X-from: beth-at-plantbio.uga.edu [mailto:beth-at-plantbio.uga.edu]
Sent: Wednesday, January 30, 2013 1:48 PM
To: pekysar-at-ucdavis.edu

Hi all,
I would like your advice/opinion on critical point dryers and sputter-
coaters. We are leaning towards the Tousimis 931 cpd (winner of the
2012 Microscopy Today Innovation Award) and the SPI Module sputter/
carbon-coaters.
thanks in advance,
Beth

==============================Original Headers==============================
2, 20 -- From beth-at-plantbio.uga.edu Wed Jan 30 15:34:52 2013
2, 20 -- Received: from dogwood.plantbio.uga.edu (pbmail.plantbio.uga.edu
[128.192.26.3])
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==============================Original Headers==============================
12, 30 -- From pekysar-at-ucdavis.edu Wed Jan 30 16:11:15 2013
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12, 30 -- To: {beth-at-plantbio.uga.edu}
12, 30 -- Cc: "Microscopy Listserver" {Microscopy-at-microscopy.com}
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We'd have to second that, we got the earlier model Tousimis 815 and it has been very good though rather greedy for CO2. It would be worth looking at the new fully automatic Leica CPD, looked good on demo. Fully automatic is the only way...

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
________________________________________
X-from: pekysar-at-ucdavis.edu [pekysar-at-ucdavis.edu]
Sent: Thursday, 31 January 2013 9:14 a.m.
To: White, Rosemary (PI, Black Mountain)

Hi Beth,
We recently purchased the Tousimis 931. I absolutely love it!!!!!!! It's
awesome. The samples I processed have been superb!!! It has tremendously
freed up my time and has been completely dependable.
I'm not familiar with the SPI sputter/carbon coaters. We have a PELCO SC-7
Sputter coater and it's probably the best one I've ever used. It's easy and
quick. However it is just a low vacuum sputter coater(0.08mbar).
Hope that helps!
Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

-----Original Message-----
X-from: beth-at-plantbio.uga.edu [mailto:beth-at-plantbio.uga.edu]
Sent: Wednesday, January 30, 2013 1:48 PM
To: pekysar-at-ucdavis.edu

Hi all,
I would like your advice/opinion on critical point dryers and sputter-
coaters. We are leaning towards the Tousimis 931 cpd (winner of the
2012 Microscopy Today Innovation Award) and the SPI Module sputter/
carbon-coaters.
thanks in advance,
Beth

==============================Original Headers==============================
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I need some details on attaching and operating a Digital camera to the Slow Scan Photography-Polaroid Fixture on an old SEM. The college doesn't seem to have enough funding for the commercial camera fixtures.
Jeff/N. Texas Area
____________________________________________________________
Woman is 57 But Looks 27
57-Year-Old Mom has a simple facelift trick that angered doctors...
http://thirdpartyoffers.juno.com/TGL3131/5109c1f678c8241f619bbst02vuc

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Dear All,
anybody out there who can help me with the correct jumper settings for the scan voltage at the NORAN SIX EDS system?
I need to have ca 8 Volt Vss for my scan-amp, now I have only 5 Vss. With the potis at the rear side I am not able to go up high
enough...

Best wishes,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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Dear Listers:

My thanks to everyone who responded to my request on flat embedding a large piece of tissue (2cm long, 1 to 1.5cm wide and 100microns thick) in LR White. So the embedding certainly looks feasible. Related to this project, the faculty member has asked me if it is possible to section the resulting block in 10 micron thick sections? This is for confocal imaging apparently. So he would like to get a section that is 10microns thick and approx. 2cm long and 1 to 1.5cm wide and I presume heat fixed to a glass slide. Now the thickest I have ever cut is 2 microns on my Diatome Jumbo Histo diamond and less than 8mm wide. Anyone out there with experience in cutting a section like this? I feel the LR White would be too brittle to go as thick as 10 microns without cracking, but I really don't know. Can the block be sectioned on a standard histology microtome, the way you section a paraffin block? Any and all advice would be appreciated.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

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Tom,

Do you have the option of embedding some of the tissue in another resin
that would not be suitable for TEM? If so than I would suggest embedding
in JB-4 resin which can be cut with larger glass knives (and also
disposable metal blades). Usually this is done on a JB-4 microtome and
uses wider glass that is cut with a special knife breaker. These pieces
of equipment may not be available to you. However, it may be possible to
use a standard paraffin microtome to cut sections of this size in this
resin. Perhaps others can comment on that since I have not tried it.

Ultrastructure of tissue embedded in JB-4 resin is of much higher quality
than that embedded in paraffin. It also can still be stained with many of
the aqueous stains used for LM.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

On 1/31/13 11:42 AM, "tbargar-at-unmc.edu" {tbargar-at-unmc.edu} wrote:

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Sent on behalf of Dr. M. K. Miller, address for queries below.

------

A new postdoc position is available immediately at Oak Ridge National Laboratory

Purpose
Under general supervision, the incumbent will focus on the nanostructural characterization of the microstructures of
nanostructured ferritic alloys under various high dose ion irradiations and thermally aged treatments. This position resides in
the Microscopy Group in the Materials Science and Technology Division (MSTD), Physical Sciences Directorate (PSD) at
Oak Ridge National Laboratory.

Major Duties/Responsibilities
- Perform atom probe tomography, scanning electron microscopy, and transmission electron microscopy
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- Prepares specimens with a dual-beam scanning electron microscope and focused ion beam miller
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- Ensures compliance with environment, safety, health and quality program requirements.
- Maintains strong commitment to the implementation and perpetuation of values and ethics.

(more details are in the attachment)

To apply:
Go to http://jobs.ornl.gov/
Select View External Positions button on the rightmost column
Enter "tomography" in the keywords and hit start button
Double clink on Postdoctoral Research Associate / NB50344514

See the buttons at the top of the page to apply.

For addition details, please e-mail (millermk-at-ornl.gov)
Dr. Michael K Miller
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Microscopy Group
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Oak Ridge, TN 37831-6139, USA

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Hi Tom,
It doesn't make sense to me to epoxy embed and cut 10um sections for
confocal examination. For confocal I suggest cutting the large tissue at
50 um then process and image with a confocal. Most confocals can image
at a depth of at least 50um. A multi-photon confocal can image deeper
~200-300um. Otherwise if the epoxy preparation is important cut 2um
sections and use a widefield microsccope--confocal is not really needed
since you already have thin sections. The individual section images can
be aligned post-microscopy. However, considering the problems of section
folding and wrinkling that I have encountered with 4mm X 4mm X 1.5mm
epoxy sections, larger sections may present even more headaches and
frustration.
Good luck,
Larry

On 1/31/2013 8:51 AM, tbargar-at-unmc.edu wrote:
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} Dear Listers:
}
} My thanks to everyone who responded to my request on flat embedding a large piece of tissue (2cm long, 1 to 1.5cm wide and 100microns thick) in LR White. So the embedding certainly looks feasible. Related to this project, the faculty member has asked me if it is possible to section the resulting block in 10 micron thick sections? This is for confocal imaging apparently. So he would like to get a section that is 10microns thick and approx. 2cm long and 1 to 1.5cm wide and I presume heat fixed to a glass slide. Now the thickest I have ever cut is 2 microns on my Diatome Jumbo Histo diamond and less than 8mm wide. Anyone out there with experience in cutting a section like this? I feel the LR White would be too brittle to go as thick as 10 microns without cracking, but I really don't know. Can the block be sectioned on a standard histology microtome, the way you section a paraffin block? Any and all advice would be appreciated.
}
} Tom Bargar
} UNMC
} Electron Microscopy Core Facility
} 986395 Nebraska Medical Center
} Omaha, NE 68198-6395
} 402-559-7347
} tbargar-at-unmc.edu
}
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--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, Diabetes & Endocrinology Research Center Microscopy Core
UCSF, Dept. of Anatomy, Rm S657
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

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415-999-4758

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Dear List
I frequently receive requests for recommendations of reliable service
providers for conventional microscopes. Issues like cleaning crusted
optics, repairing and replacing springs etc. Are there any you can
recommend that work in Southern California, USA?

Thanks
Mary

--
Mary Raven
Microscopy Facility Director
http://www.lifesci.ucsb.edu/~m_raven/
Phone: (805) 893 8702

Neuroscience Research Institute &
Molecular, Cellular & Developmental Biology
Biology 2 Building, room 5173
The University of California
Santa Barbara, CA 93106-5060

2014 Advanced Microscopy and Digital Imaging Workshop
https://www.lifesci.ucsb.edu/mcdb/events/imaging_workshop/
Jan 14- Jan 17, 2014

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Hello,
We would like to invite you to an ultra microtomy work shop hosted by
San Joaquin Delta College, Stockton, on February 25th-28th. We will
cover both room temp and cryo processes for material and biological
specimen sample prep. There will be lectures in the morning followed by
"hands-on" training after lunch. We encourage our attendees to bring a
sample of what they are currently working on. SJDC - Stockton has a well
equipped imaging facility and RMC will have the latest cryo
ultramicrotome and glass knife maker to use. There is no charge to
attend the workshop, lunch is included, but attendees will be
responsible for their own travel and accomodation expenses. Space is
limited so please let me know as soon as possible. February 7th is the
sigh up deadline.
Thank you,
Mike Leeper

--
Mike Leeper
RMC National Sales and Service Manager
Boeckeler Instruments, Inc.
4650 S. Butterfield Drive
Tucson, AZ U.S.A. 85714
Phone: (520) 745-0001 Fax: (520) 745-0004

mike-at-boeckeler.com
www.rmcproducts.com

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Email: margaret.dienelt-at-ars.usda.gov
Name: Margaret Dienelt

Organization: Agricultural Research Service, USDA

Title-Subject: [Filtered] Need manual for EM Tech K850 Critical point dryer

Message: Hello Everyone,

We have an old EM Tech K850 critical point dryer in our lab that has never been used. After all these years, we need to use it but of course the manual is nowhere to be found. Does anyone have a manual to this instrument they can spare? If you have a manual but can't spare it, would you please share the contact information of EM Tech with us?

Another option: if you can spare your manual for a week, we will copy it at our expense and send it back the next day.

Thank you very much.

Margaret Dienelt
Plant Pathologist/Electron Microscopist
Floral and Nursery Plants Research Unit
NA/ARS/USDA

Bldg 012 Rm 1-7
10300 Baltimore Avenue
Beltsville, MD 20705 USA

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I haven't used Spurr's for many years. I use Eponate 812 or EMbed--Epon
equivalents. I put a drop of water on a glass slide then transfer a
section from the knife boat to the drop of water with a Minutien pin
(very fine needle) glued on a wood applicator stick. Usually I put two
or three sections on a drop but for larger sections one per drop. The
slide then goes on a hot plate set at a temperature so the water drop
dries slowly ~3-4 minutes. Sometimes just allowing the water to air dry
at room temperature works better. Then I stain the sections with
Toluidine Blue for 10-20 seconds and rinse with water. I am usually
just looking for an area that I wish to thin section and do not use the
1-2um sections for microscopy. Another possibility to minimize wrinkles
is to stretch the sections in the knife boat--with a heat pen or solvent
vapors (chloroform vapors are not healthy).
Happy Sectioning!
Larry

On 1/31/2013 12:52 PM, Philip Oshel wrote:
} Larry,
}
} How do you deal with section wrinkling & folding in large sections?
} I've got a student do LM on corn kernels - embryonic development - and
} he's reached a stage where he needs large area sections. Not 4x4 mm,
} but a couple of millimeters, maybe more. Using Spurr's, I believe.
} That's what I have, and I suspect the prof he's working with is also
} using that, not some acrylate or the like.
} This is for LM and tol-blue staining, although they're moving towards
} doing some TEM.
}
} Phil
}
} }
} } Hi Tom,
} } It doesn't make sense to me to epoxy embed and cut 10um sections for
} } confocal examination. For confocal I suggest cutting the large tissue at
} } 50 um then process and image with a confocal. Most confocals can image
} } at a depth of at least 50um. A multi-photon confocal can image deeper
} } ~200-300um. Otherwise if the epoxy preparation is important cut 2um
} } sections and use a widefield microsccope--confocal is not really needed
} } since you already have thin sections. The individual section images can
} } be aligned post-microscopy. However, considering the problems of section
} } folding and wrinkling that I have encountered with 4mm X 4mm X 1.5mm
} } epoxy sections, larger sections may present even more headaches and
} } frustration.
} } Good luck,
} } Larry
}

--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, Diabetes & Endocrinology Research Center Microscopy Core
UCSF, Dept. of Anatomy, Rm S657
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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Dear Colleagues,

The 3DEM GRC in 2014 will take place from June 22 to 27, 2014, in Girona, Spain. This is close to Barcelona.

Please mark your calendars.

Henning.

Henning Stahlberg, PhD
Prof. for Structural Biology, C-CINA, Biozentrum, University Basel
Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland
http://c-cina.org | Tel. +41-61-387 32 62 | mailto:Henning.Stahlberg-at-unibas.ch

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10, 31 -- Subject: 3DEM GRC 2014 will be June 22-27 at Melia Golf Vichy Catalan,
10, 31 -- Girona, Spain
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Scientific Services at The Jackson Laboratory is conducting benchmarking surveys in an effort to develop an understanding of the current best practices in operations, management and technical delivery at peer institutions. The goal is to gather quality data that can be used to assist us and all survey participants with operational assessment and planning. Survey participants will receive compiled results which will be de-identified before distribution. Thank you in advance for your participation.

Pete Finger
Manager, Electron Microscopy Service
The Jackson Laboratory
Bar Harbor, ME 04609
207-288-6337

"Do your duty in all things. You can not do more, you should never wish to do less".
- Robert E. Lee

The information in this email, including attachments, may be confidential and is intended solely for the addressee(s). If you believe you received this email by mistake, please notify the sender by return email as soon as possible.

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The NUANCE Center at Northwestern University is going to be producing a new web series called "What's It Made Of?" It will be a short, semi-educational show mainly featuring SEM and EDS to examine normal, everyday objects. We'd love to use this as a fun educational outreach to show the uninitiated the types of things that we can do in the world of electron microscopy. So please share with all your friends and whoever.

The first episode has been posted and examines the sparkly ink on the $20 bill.

http://youtu.be/z3qHg1QOAk4

ERiC Jay Miller
Microscopy & Imaging Specialist
Electron Probe Instrumentation Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1152 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 467-0789
fax: (847) 467-657

http://www.nuance.northwestern.edu

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Dear Collective,

Might someone out there have manuals for a Gatan 601 Ultrasonic Disc Cutter=
and Gatan 656 Dimple Grinder that they would be willing to share, copy, le=
nd, or PDF? We have two lonely instruments waiting for some attention, but=
we just don't know their proper care and feeding, yet. Any assistance wou=
ld be greatly appreciated and postage, copying, etc. costs reimbursed with =
all dispatch.

Thanks!

Randy
(from home----signature's on the office computer----too lazy to add one her=
e now)

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4, 27 -- From tindallr-at-missouri.edu Fri Feb 8 16:44:43 2013
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4, 27 -- Subject: Gatan manuals?
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Thanks to all who responded to our request for manuals. We have either received the ones we need, or they are on their way. What a great bunch you guys are!

Randy

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Dear Collective,

I know. I'm getting to be a real pest, but the day of reckoning has come. We need to take sequential sections down through an HPF-frozen sample and doubt that we will be able to get the sample out of the cup. This means we will probably have to trim away the cup from around the sample. We know this can be done (in fact, we are trimming away gold with an old beat-up diamond as I write this), but doing it in a time frame that makes it affordable and doesn't destroy the knife in the process is the problem.

A couple potential approaches:

1) pretrimming the block with a dremel or similar tool and leaving a small amount of gold intact around the sample cup area before doing HPF. Would this still provide an adequate seal during the actual freezing?

2) doing the trimming of the metal in a cryostat with a steel knife after freezing the sample, then transferring into the cryo-ultramicrotome for the final sections.

Does anyone have any thoughts on these or other approaches? To repeat, we hope to get the cup-trimming part down to a manageable amount of time, without trashing every (expensive) knife we have.

Thanks!

Cheers,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

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13, 30 -- CC: "White, Tommi A." {whiteto-at-missouri.edu} ,
13, 30 -- "Vinson, Juliana Pavlovna"
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I doubt a cryostat would be cold enough to prevent crystallization of the of vitrified ice.

If a diamond can cut the gold, have you tried a glass knife?

Good luck and please share the final method that leads to success. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: tindallr-at-missouri.edu [mailto:tindallr-at-missouri.edu]
Sent: Monday, February 11, 2013 10:11 AM
To: Phillips, Thomas E.

Dear Collective,

I know. I'm getting to be a real pest, but the day of reckoning has come. We need to take sequential sections down through an HPF-frozen sample and doubt that we will be able to get the sample out of the cup. This means we will probably have to trim away the cup from around the sample. We know this can be done (in fact, we are trimming away gold with an old beat-up diamond as I write this), but doing it in a time frame that makes it affordable and doesn't destroy the knife in the process is the problem.

A couple potential approaches:

1) pretrimming the block with a dremel or similar tool and leaving a small amount of gold intact around the sample cup area before doing HPF. Would this still provide an adequate seal during the actual freezing?

2) doing the trimming of the metal in a cryostat with a steel knife after freezing the sample, then transferring into the cryo-ultramicrotome for the final sections.

Does anyone have any thoughts on these or other approaches? To repeat, we hope to get the cup-trimming part down to a manageable amount of time, without trashing every (expensive) knife we have.

Thanks!

Cheers,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

==============================Original Headers==============================
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13, 30 -- "Vinson, Juliana Pavlovna"
13, 30 -- {VinsonJu-at-missouri.edu}
13, 30 -- Subject: Cryo-ultramicrotomy: trimming hpf cups 13, 30 -- Thread-Topic: Cryo-ultramicrotomy: trimming hpf cups 13, 30 -- Thread-Index: Ac4IcK0CNxNUfEndSeah6b2Z4NAYDQ== 13, 30 -- Date: Mon, 11 Feb 2013 16:10:25 +0000 13, 30 -- Message-ID: {995EF3174193844182C7E47258DCB04C5B76EC-at-UM-MBX-N03.um.umsystem.edu}
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25, 31 -- From PhillipsT-at-missouri.edu Mon Feb 11 10:35:58 2013
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25, 31 -- Subject: RE: [Microscopy] Cryo-ultramicrotomy: trimming hpf cups
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I just remembered - I think the cryo-ultramicrotome has a tool for this! It is a metal rod with a sharp end that fits in the knife holder - take a peek in your goodies box. you could have extras made at the machine shop

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

--------------------------------------
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I doubt a cryostat would be cold enough to prevent crystallization of the of vitrified ice.

If a diamond can cut the gold, have you tried a glass knife?

Good luck and please share the final method that leads to success. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: tindallr-at-missouri.edu [mailto:tindallr-at-missouri.edu]
Sent: Monday, February 11, 2013 10:11 AM
To: Phillips, Thomas E.

Dear Collective,

I know. I'm getting to be a real pest, but the day of reckoning has come. We need to take sequential sections down through an HPF-frozen sample and doubt that we will be able to get the sample out of the cup. This means we will probably have to trim away the cup from around the sample. We know this can be done (in fact, we are trimming away gold with an old beat-up diamond as I write this), but doing it in a time frame that makes it affordable and doesn't destroy the knife in the process is the problem.

A couple potential approaches:

1) pretrimming the block with a dremel or similar tool and leaving a small amount of gold intact around the sample cup area before doing HPF. Would this still provide an adequate seal during the actual freezing?

2) doing the trimming of the metal in a cryostat with a steel knife after freezing the sample, then transferring into the cryo-ultramicrotome for the final sections.

Does anyone have any thoughts on these or other approaches? To repeat, we hope to get the cup-trimming part down to a manageable amount of time, without trashing every (expensive) knife we have.

Thanks!

Cheers,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

==============================Original Headers==============================
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13, 30 -- "Vinson, Juliana Pavlovna"
13, 30 -- {VinsonJu-at-missouri.edu}
13, 30 -- Subject: Cryo-ultramicrotomy: trimming hpf cups 13, 30 -- Thread-Topic: Cryo-ultramicrotomy: trimming hpf cups 13, 30 -- Thread-Index: Ac4IcK0CNxNUfEndSeah6b2Z4NAYDQ== 13, 30 -- Date: Mon, 11 Feb 2013 16:10:25 +0000 13, 30 -- Message-ID: {995EF3174193844182C7E47258DCB04C5B76EC-at-UM-MBX-N03.um.umsystem.edu}
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25, 31 -- Subject: RE: [Microscopy] Cryo-ultramicrotomy: trimming hpf cups
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As Tom Phillips suggests, there are options in the cryoultramicrotome. We do trimming of frozen roots with attached soil with a tungsten-coated glass knife, which would probably work for you.

Also, could you make/buy the freezing cups in a softer metal or other material?

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
________________________________________
X-from: tindallr-at-missouri.edu [tindallr-at-missouri.edu]
Sent: Tuesday, 12 February 2013 3:17 a.m.
To: White, Rosemary (PI, Black Mountain)

Dear Collective,

I know. I'm getting to be a real pest, but the day of reckoning has come. We need to take sequential sections down through an HPF-frozen sample and doubt that we will be able to get the sample out of the cup. This means we will probably have to trim away the cup from around the sample. We know this can be done (in fact, we are trimming away gold with an old beat-up diamond as I write this), but doing it in a time frame that makes it affordable and doesn't destroy the knife in the process is the problem.

A couple potential approaches:

1) pretrimming the block with a dremel or similar tool and leaving a small amount of gold intact around the sample cup area before doing HPF. Would this still provide an adequate seal during the actual freezing?

2) doing the trimming of the metal in a cryostat with a steel knife after freezing the sample, then transferring into the cryo-ultramicrotome for the final sections.

Does anyone have any thoughts on these or other approaches? To repeat, we hope to get the cup-trimming part down to a manageable amount of time, without trashing every (expensive) knife we have.

Thanks!

Cheers,
Randy

Randy Tindall
Senior Research Specialist
Electron Microscopy Core Facility
W125 Veterinary Medicine
University of Missouri
Columbia, MO 65201
(573) 884-5383

We Do Small Well!

www.emc.missouri.edu (dayjob)
http://nadiasyard.com (blog)
http://www.facebook.com/RandyTindallGalleries (photography)

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From noreply-at-creatividentro.com Mon Feb 11 13:54:07 2013
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Hello,

I am having an issue with the moveable aperture (part MP-30060) on my
JSM-6490LV and am hoping someone out there has had a similar issue and can
be kind enough to provide some insight.

The mechanism has coarse positioning mechanism to hold the objective
aperture strip in three positions plus a fully extracted position.
Currently, it is only able to be positioned in the middle two positions
(aperture positions #2, and #1). Any help would be appreciated.

Thanks in advance,
Bryan

--
_________________________________________________________________
Bryan R. Bandli

Research Instrumentation Laboratory Manager
University of Minnesota, Duluth
229 Heller Hall
1114 Kirby Dr.
Duluth, MN 55812
218-726-7362
==================================================================

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My experience with JEOL (and many other) aperture carriers is that the fine adjustment is enough to (nearly) shift from one aperture to another. So instead of seeing apertures 1, 2, 3, and OPEN you end up seeing nothing, 1, 2, and OPEN or 2, 3, nothing, and OPEN. If you have an ammeter on your system or an EDS system, you can check eventually determine which situation you have. You would have to know what the normal current or x-ray signal is for a given aperture and condenser lens setting.

I would experiment, carefully, with the longitudinal adjustment to see of you can bring the next aperture into view. You should then find all three apertures in their usual places.

BTW, I had trouble with a user damaging our strip. I tried to adjust my x-ray count rate but there was no difference between the positions. It turned out that they forgot the order of the twist and pull operations, and they put a 45-degree twist in the aperture strip. They dropped the strip down into the column. At lease it did not block the beam. Your problem is considerable simpler

Warren.
________________________________________
X-from: bbandli-at-d.umn.edu [bbandli-at-d.umn.edu]
Sent: Monday, February 11, 2013 2:37 PM
To: wesaia-at-iastate.edu

Hello,

I am having an issue with the moveable aperture (part MP-30060) on my
JSM-6490LV and am hoping someone out there has had a similar issue and can
be kind enough to provide some insight.

The mechanism has coarse positioning mechanism to hold the objective
aperture strip in three positions plus a fully extracted position.
Currently, it is only able to be positioned in the middle two positions
(aperture positions #2, and #1). Any help would be appreciated.

Thanks in advance,
Bryan

--
_________________________________________________________________
Bryan R. Bandli

Research Instrumentation Laboratory Manager
University of Minnesota, Duluth
229 Heller Hall
1114 Kirby Dr.
Duluth, MN 55812
218-726-7362
==================================================================

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14, 39 -- Subject: RE: [Microscopy] JEOL aperture problem
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Good Afternoon,

Does anyone have any tips for hardening some epoxy blocks that are too soft to section? Somehow, they didn�t harden as well as they usually do. I�ve put them back in the oven for a longer period of time, and they did get a bit harder, but not enough. They �bend�!! This has happened to me only twice in over 23 years! I�d really like to be able to section the samples (Fly embryos) for the PI.

Thanks,
Virginia Crocker
Biologist, NIH EM Facility

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As the deadline for abstract submission approaches and plans are being
made to attend the Indianapolis meetings Aug. 4-8, please don*t forget
about MSA*s student bursary program. Its purpose is to encourage
students to attend the meetings by helping to defray some of the costs
and giving them an opportunity to meet and interact with the established
microscopy community.

The students will be paid $10 an hour to work for ~20 hours (up to 40
hours) during the meeting or pre-meeting events. The jobs involve such
things as providing support in the different symposia (assisting with
audio-visual needs, speaker set-up, maintaining an attendance count),
staffing the volunteer office, monitoring use of the Internet Caf�, and
helping with vendor tutorial sign-up. Payment is given as a check at
the end of the meetings or when the student leaves Indianapolis.

Once the program has been finalized, each registered bursary will be
contacted and allowed to choose the times and activities they would like
to work. Many times they end up *working* sessions they would
attend anyway. There is an added bonus of $10 cash to help with meals
for each morning and/or afternoon session worked and a meeting shirt.

If anyone would like to participate in the bursary program, please
check the *I wish to apply for a student bursary* box in section 2
of the registration form. Bursary space is limited, so sign-up early.
Applicants for the bursaries must be members of MSA or MAS, enrolled as
students at a recognized educational institution, and have paid their
registration fee.

For those *non-students* volunteers are also needed to help with
the above mentioned meeting activities. Although not paid on an hourly
basis as the student bursaries, volunteers do receive a meeting shirt
and the same cash allotment to help with meals. They also have the
opportunity to interact more with the microscopy community as they
assist with meeting tasks.

If anyone has any questions about the bursary/volunteer program, or
would like to participate, please contact:

Amanda Lawrence
Institute for Imaging and Analytical Technologies
Mississippi State University
662-325-7998
alawrence-at-i2at.msstate.edu

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Email: kenhon-at-hawaii.edu
Name: Ken Hon

Organization: University of Hawaii at Hilo

Title-Subject: [Filtered] Student Petrographic Microscope

Message: Aloha,

We are thinking about slowly replacing our aging Leitz monocular student
petrographic microscopes used in mineralogy and petrology. I was
wondering if anyone had suggestions for relatively inexpensive, but
durable binocular petrographic microscopes for use by students. Also,
they must withstand relatively high humidity. Even in a cabinet with a
dehumidifier stick in an air conditioned lab, the lowest we can get
humidity is about 50%. The Leitz's have lasted over 20 years and are
still workable, so they've been great from that standpoint. However,
they no longer make parts for them and we can't get them serviced any
longer. So, I'd welcome any ideas and suggestions for dealers (if
that's allowed on this board).

Aloha,

Ken

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Email: IMAGE-at-siu.edu
Name: George Trifon

Organization: Southern Illinois University

Title-Subject: [Filtered] DIRECTOR, INTEGRATED MICROSCOPY & GRAPHICS
EXPERTISE (IMAGE) FACILITY

Message: DIRECTOR, INTEGRATED MICROSCOPY & GRAPHICS EXPERTISE (IMAGE)
FACILITY
(Deadline Extended - 12 mo. Term, A/P)

Southern Illinois University Carbondale seeks to hire a Director for the
Integrated Microscopy & Graphic Expertise Facility. The Director will be
responsible for operating and maintaining state-of-the-art microscopy
equipment, teaching SEM and TEM classes, printing large posters for
research presentations, generating external contracts and grants,
overseeing billing and inventory control, and engaging in outreach and
promotional activities across campus and in the community.

Minimum Qualifications: Applicants must have a Masters degree in
Science or Engineering and four (4) years experience as a professional
electron microscopist in a centralized, service-oriented facility with
multidisciplinary clients.

Preferred Qualifications: Experience in trouble-shooting with advanced
instrumentation. A working knowledge of graphic programs for use in
professional publications and large format posters. The ability to work
independently with minimal administrative oversight or support.
Excellent organizational and communication skills. Ph.D. preferred.

Website: image.siu.edu

Extended Deadline: Application review will begin Feb 18, 2013, and
continue until filled

Application: Send letter of application, resume, and three reference

letters via the website or hard copy to:

Chair, Search Committee
Office of Sponsored Projects Administration
SIU Carbondale
900 S Normal, Woody Hall C206, MC 4709
Carbondale, IL 62901

SIU Carbondale is an affirmative action/equal opportunity employer that
strives to enhance its ability to develop a diverse faculty and staff
and to increase its potential to serve a diverse student population. All
applications are welcomed and encouraged and will receive consideration.

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Email: serene_ng-at-dsi.a-star.edu.sg
Name: Serene

Title-Subject: [Filtered] Electron microscopy school

Message: Dear Listers,

I am looking for electron microscopy school(s) which offer short or
full-term courses (TEM in particular) - both theory and practical
lessons will be good.

I know there's Lehigh and Hooke which offer short courses. Are there
other institutions fellow listers can suggest? Are there similar schools
hosted in Europe or
Asia?

Thank you listers!
Serene

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Email: amit.welcomes.u-at-gmail.com
Name: Amit

Organization: IISc

Title-Subject: [Filtered] CCD camera sensor size

Message: Good evening,
I was wondering if anyone knows any way to find out exact sensor size of
ccd camera? (i want to measure actual distance, like in films, between
diffraction spots, any other suggestions are also welcome!)
i read the brochure of manufacturer, many things dont make sense there,
e.g chip size- 2/3 inches, 10.2mmx8.3mm
but wont 10.3mm x 8.3 mm be 1.31 cm diagonally? while 2/3 inches should
be 1.69 cm.
pixcel count-1376x 1032

thank you

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Email: trogadisj-at-smh.ca
Name: Judy Trogadis

Organization: St. Michael's Hospital

Title-Subject: [Filtered] microtome accuracy

Message: Fellow microscopists

We are planning to do very fine optical measurements from tissue
sections, therefore, control of thickness and of even surface is critical.

Which are the most accurate and reproducible microtomes on the market
for paraffin or frozen blocks? Are there laser based tissue sectioning
equipment.

Any suggestions are welcomed, including replies from manufacturers.
Thank you,
Judy

Judy Trogadis
Bio-Imaging Coordinator
Keenan Research Centre, St. Michael's
209 Victoria Street
Toronto M5B 1T8, Canada
office: 416-864-6060 ext. 77612
imaging facility: ext. 77434
cell: 416-254-9330
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Email: yankova-at-neuron.uchc.edu
Name: Maya Yankova

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Title-Subject: [Filtered] LKB Manual needed

Message: Hello,
A researcher in our lab inherited an LKB microtome model 8800, but the
manual is missing. We were wandering if anyone has a pdf or hard copy
they could send us.

Thank you,

Maya Yankova
University of Connecticut,Health center

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Title-Subject: [Filtered] Refurbished DoubleTilt Holder

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Email: karen.rothberg-at-utsouthwestern.edu
Name: Karen Rothberg

Organization: UT Southwestern Medical Center

Title-Subject: [Filtered] Fixation and Processing of Mouse Dorsal Root
Ganglia

Message: Hi,

We have a user who wants 0.5 micron cross sections of the L3 ganglia
and spinal chords from mice. What is the best way to process the tissue
without mechanical damage to the root?

They did perfuse the mouse with Paraformaldehye/Glut and gave the
samples to us in 2.5% glut. They dissected the ganglia from the spinal
chord and we put the roots in agarose prior to osmium. They do not want
to do electron microscopy on the tissue but need the resin embedding.

The second thing they want to do is quantitate the Tol blue staining
from the thick sections. Is that doable and reproducible?

thanks,
Karen
UT Southwestern Medical Center
Live Cell and Electron Microcopy Core Facilities
214-648-2805

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Title-Subject: [Filtered] sem

Message: can i insert wood in electron scanning microscope quanta 200

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Name: Ian MacLaren

Organization: University of Glasgow / EMAG (Institute of Physics)

Title-Subject: [Filtered] EMAG 2013 - Call for abstracts - deadline 22nd
March 2013

Message: Electron Microscopy and Analysis Group Conference 2013

Organised by the IOP Electron Microscopy and Analysis Group

www.emag-iop.org

3 6 September 2013

University of York, York, UK

Conference highlights

High profile plenary speakers
Prof. Ahmed Zewail (Nobel Prize in Chemistry 1999, CalTech)
Prof. Archie Howie (University of Cambridge)
Prof. Pratibha Gai (University of York)

Special symposium on in-situ microscopy in honour of Prof. Archie Howie

Refereed proceedings

Short courses on advanced topics in electron microscopy (Monday 2 and
Tuesday 3 September)
Aberration-corrected electron microscopy
Spectroscopy using electrons and X-rays in the electron microscope
Simulation of TEM and STEM electron microscope images

Trade exhibition

Call for abstracts

We invite abstract submissions for oral or poster presentations on any
of the following topics:

Technique developments and their applications:
The Archie Howie symposium on in-situ microscopy
Advances in electron microscope imaging
Spectroscopy and analysis
Developments in STEM
Modelling and quantification of electron microscope data
Applications of electron microscopy to specific areas:
Nanomaterials
Structural materials
Functional materials
Biomaterials and hard-soft matter interfaces in biomedical and
environmental science

Key dates

Abstract submission deadline: 22 March 2013
Early registration deadline: 2 August 2013

Abstracts should be submitted online at www.emag-iop.org

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There should be no problem examining wood in the Quanta as far as the SEM is concerned. The question will be what changed take place in the sample and if they are acceptable.

You might need to use environmental mode if it is essential to keep the moisture content the same. Otherwise, you could use variable pressure mode to eliminate charging without coating.

What are you looking for in your sample that you want to use SEM in the first place?

Warren Straszheim
Materials Analysis Lab, Iowa State University
home of a Quanta 250 FEG and other equipment

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Email: hadeelzh-at-yahoo.com
Name: hadeel

Organization: yarmouk university

Title-Subject: [Filtered] sem

Message: can i insert wood in electron scanning microscope quanta 200

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Here is one:

http://www.edmundoptics.com/learning-and-support/technical/learning-center/application-notes/imaging/electronic-imaging-resource-guide/?&pagenum=5#4.3

google "CCD size" for more

Vitaly Feingold
SIA
2773 Heath Lane
Duluth GA 30096
Ph. 770-232-7785
Fax 770-232-1791
www.sia-cam.com
vitaly-at-sia-cam.com

On 2/12/2013 11:15 PM, microscopylistserver-noreply-at-microscopy.com wrote:
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} Name: Amit
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} Title-Subject: [Filtered] CCD camera sensor size
}
} Message: Good evening,
} I was wondering if anyone knows any way to find out exact sensor size of
} ccd camera? (i want to measure actual distance, like in films, between
} diffraction spots, any other suggestions are also welcome!)
} i read the brochure of manufacturer, many things dont make sense there,
} e.g chip size- 2/3 inches, 10.2mmx8.3mm
} but wont 10.3mm x 8.3 mm be 1.31 cm diagonally? while 2/3 inches should
} be 1.69 cm.
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Name: Rossou Ludo

Organization: University of Antwerp EMAT group (Belgium)

Title-Subject: [Filtered] sample holder for Baltec RES 010

Message: Dear all,

I am looking for sample holders for the old Baltec RES 010. In our lab,
a few are still working.
I think that it can be that somewhere this ion milling machines are not
in use anymore.
So I am very interested in all kind of holders.

Contact me directly.

Thanks in advance,
Ludo

Rossou Ludo
Universiteit Antwerpen
Departement Natuurkunde
Dienst EMAT
Groeneborgerlaan 171
B-2020 Antwerpen
Belgium
Tel. 00 32 3 265 32 55
Fax.00 32 3 265 33 18
ludo.rossou-at-ua.ac.be
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Serene,

Central Michigan University.
We have a "microscopy major" - a microscopy option within the biology
major. The courses are light microscopy, scanning laser confocal
microscopy, transmission electron microscopy, and scanning electron
microscopy (with EDS if needed by the student). They all cover both
theory and practical lab work, and are full semester courses leading to
a B.Sc.
The courses are also open to non-majors, so we commonly have materials
science (undergrad and PhD students) and geology students..

Phil

} Email: serene_ng-at-dsi.a-star.edu.sg
} Name: Serene
}
} Title-Subject: [Filtered] Electron microscopy school
}
} Message: Dear Listers,
}
} I am looking for electron microscopy school(s) which offer short or
} full-term courses (TEM in particular) - both theory and practical
} lessons will be good.
}
} I know there's Lehigh and Hooke which offer short courses. Are there
} other institutions fellow listers can suggest? Are there similar schools
} hosted in Europe or
} Asia?
}
} Thank you listers!
} Serene

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Back in January I posted a question about using a Nikon 1 series
inter-changeable lens, small sensor camera for microscopy.

These cameras have a 1" sensor (the recent thread on CCD sizes reminded me
to get back to the list with my experiences), which is ideal for this
application.

I bought a Nikon V1, which is heavily discounted at the moment, less than
30% of the new RRP: �250 with lens.
We also bought a c-mount to Nikon 1 series adapter, IR remote release,
fast SD card and card reader, mini-HDMI to HDMI cable, and a cheap full HD
monitor with HDMI input (total cost under �140).

So for under �400 GBP we have a complete imaging system.

Some photos of the set-up:

http://booking.mrc.ox.ac.uk/NikonV1_microscope_mount.jpg

We fitted this to a Leitz Dialux microscope (160mm tube length) with a 1x
c-mount adapter on the trinocular head's photo port. The C-mount adapter
gives perfect focus registration with the image in the oculars.

The shutter is fired by IR remote release. After turning on the camera, it
takes three button presses to engage the mode where it waits for the IR
signal. All other settings (e.g. Exposure mode, white balance and ISO) are
saved when the camera is turned off. The camera will only work in fully
manual exposure (the lenses normally communicate electronically with the
body, and without this, the camera isn't very helpful). The live view on
the monitor doesn't show the effect of changing the exposure, it is always
auto-gained. It does show white balance effects though. Having said that,
as it will do a quick review of the shot after it is taken, and changing
the shutter speed is just a matter of nudging a lever on the back of the
camera, if only takes a few shots to get the right exposure.

We are using the live view on a 20" LCD via HDMI, which has very fast
frame rate and is lag-free for focussing and searching.

The camera does show the imperfections in the peri-plan optics of the
low-end EF objectives, but cropping to the central region gets rid of the
soft edges.

E.g. This example taken with the 10x 0.25NA EF objective:
http://booking.mrc.ox.ac.uk/exampleV1image.jpg

The camera can also taken impressive full HD movies at 60fps, so would be
useful for teaching purposes.

Hope this mini-review is of use to someone looking for colour brightfield
imaging on a low budget.

Ben

--
Research Support Manager
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271649
{http://www.mrc.ox.ac.uk/}

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Anyone out there using the SEM mounted microCT scanners? Would like to discuss real world practicality and use examples as my Museum considers the future of CT going forward and this might solve a dilemma we are having in addressing the range of sample sizes and resolutions.

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012�� MRC104
Washington DC 20013-7012
202-633-0891

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Hi Amit

Forgive me if I have missed something obvious since I only have experience with biological specimens and 1 vendor's software.

My understanding is that if the images is calibrated then the distance can be calculated from the image using software. The number of pixels x the distance per pixel gives the measured distance: measurement software can do this for you, you just select the 2 points. If the images you speak of is acquired on a CCD camera, then it should be (or can be) calibrated, and the camera software may allow the measurements you require.

If you are attempting to measure distance on an image that has been exported from that software since acquisition then care should be taken since you may not have the original correct calibration information associated with the image. Keep in mind such things as binning, changes to pixel size/shape etc on post processed images.

There are others on the list with more experience but hope this helps.
Naomi

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Here is one:

http://www.edmundoptics.com/learning-and-support/technical/learning-center/application-notes/imaging/electronic-imaging-resource-guide/?&pagenum=5#4.3

google "CCD size" for more

Vitaly Feingold
SIA
2773 Heath Lane
Duluth GA 30096
Ph. 770-232-7785
Fax 770-232-1791
www.sia-cam.com
vitaly-at-sia-cam.com

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} Title-Subject: [Filtered] CCD camera sensor size
}
} Message: Good evening,
} I was wondering if anyone knows any way to find out exact sensor size of
} ccd camera? (i want to measure actual distance, like in films, between
} diffraction spots, any other suggestions are also welcome!)
} i read the brochure of manufacturer, many things dont make sense there,
} e.g chip size- 2/3 inches, 10.2mmx8.3mm
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19, 26 -- From prvs=17571c23d3=Naomi_McCallum-at-health.qld.gov.au Wed Feb 13 20:33:12 2013
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Hi Judy

Unfortunately we don't have much detail on what (size) you will be attempting to measure and thus degree of accuracy that you are seeking. However, even with the "most accurate" microtome on the market, a lot can happen from tissue dissection through processing/freezing, and especially during & after microtomy that can affect accuracy .

Here's an idea from left field: a researcher engaging our services injected fluorescent polystyrene beads (commercial name escapes me at present) into their specimens to locate area of interest. (Excellent for accurate sampling and dissection.) When processed, the beads dissolved but left replicas as perfect resin circles within the tissue (ie in the fat adjacent to the area of interest.

If this was applied to frozen section, the beads would not dissolve if aqueously mounted, and the fluorescent label would allow localisation of the beads for measurement calibration. Or if paraffin processing was employed it would be the same as we experienced with the resin, you have to look for the regular spherical spaces where the beads once were.

If this is not helpful, feel free to file appropriately.

Kind regards
Naomi

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Email: trogadisj-at-smh.ca
Name: Judy Trogadis

Organization: St. Michael's Hospital

Title-Subject: [Filtered] microtome accuracy

Message: Fellow microscopists

We are planning to do very fine optical measurements from tissue
sections, therefore, control of thickness and of even surface is critical.

Which are the most accurate and reproducible microtomes on the market
for paraffin or frozen blocks? Are there laser based tissue sectioning
equipment.

Any suggestions are welcomed, including replies from manufacturers.
Thank you,
Judy

Judy Trogadis
Bio-Imaging Coordinator
Keenan Research Centre, St. Michael's
209 Victoria Street
Toronto M5B 1T8, Canada
office: 416-864-6060 ext. 77612
imaging facility: ext. 77434
cell: 416-254-9330
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Email: kalennon-at-hagerstowncc.edu
Name: Kristen Lennon

Organization: Hagerstown Community College

Title-Subject: [Filtered] microscopy outreach - microscopes

Message: Hello Everyone,

I've just been given the exciting and daunting challenge of creating a
K-12 outreach and teacher education program centered on microscopy.I've
visited the Project Micro web site and am using those resources, but I'm
wondering if any of you have any other suggested resources. I'd also
like some suggestions as to where to buy good quality stereo microscopes
at a K-12 non-profit kind of price. The idea is to get more kids excited
about science through the engaging microscopic world and make microscopy
more accessible to K-12 students and their teachers.

Your advice is, as always, much appreciated.
Best,
Kristen Lennon
kalennon-at-hagerstowncc.edu

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Thank you for replying first of all.
Thank you Vitaly Feingold for the link it helped a lot in clearing up
jargon. Though no solutions were given!

Naomi McCallum: Actually i want to measure varios constants of microscope
for example exact magnification and spherical aberration etc. for that what
we require is how much the image feature has actually "moved". earlier when
films were used it was easy as you can take out the films and measure it
with a foot ruler or something(thats what is sounds like from papers i have
read, if i am missing something then sorry, may be someone more experienced
might correct me). but to find out that now i need what is size of each
pixel.
software allots size of each pixel according to values given to it (eg.0466
nm /pixel etc). but actual pixel size is in micrometers, i.e physical size
of sensor on camera. To avoid binning etc i am making sure that camera runs
on maximum resolution.

I have tried following method but dont know if its correct.-
1. obtain diffraction pattern of some single crystal sample
2.increase the camera length to magnify it
3.fluorescent screens have markers on them (jeol one that we are using has
5mm ticks at the lower side)
4. move any spot on that tticks and take images.
5. you can get physical distance from that ticks(ie. 5mm) and number of
pixels moved in image.
we can then find out whats the exact physical size of pixel

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4, 28 -- From amit.welcomes.u-at-gmail.com Thu Feb 14 03:27:36 2013
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Dear all,

We are pleased to announce

the 3rd UB Practical Course on Cryo-Electron Microscopy of Vitreous
Sections (CEMOVIS, a.k.a cryoEM of frozen-hydrated sections)

2-4 July 2013 / 9-11 July 2013

Organiser : Beno�t Zuber
Instructors: Beno�t Zuber, Daniel Studer, Ioan Iacovache

The objective of the course is to teach participants the practical
skills necessary to successfully apply CEMOVIS (a.k.a. Frozen-hydrated
sections) in their laboratories. Our latest tricks making CEMOVIS
quite easy will be demonstrated. Essential background theory of
CEMOVIS will be given and most of the time will be spent practicing
high-pressure freezing, cryo-sectioning, and low-dose TEM imaging of
vitreous sections. 1 high-pressure freezing machine, 3
state-of-the-art cryo-ultramicrotomes and 1 cryo-electron microscope
will be dedicated to the course. The 3-day course is given twice for
two different groups of maximum 4 participants each so that every
participant can be actively practicing during the whole course.

All the participants of the previous previous courses have been
successful in producing, collecting and imaging vitreous sections.

The course is intended for scientists whose research projects will
benefit from the use of CEMOVIS. Experience in either cryo-electron
microscopy or ultramicrotomy of resin-embedded specimens is a
prerequisite.

Information and registration:
http://www.ana.unibe.ch/events/cemovis/index.html

Registration deadline: 17.03.2013

Yours faithfully
Beno�t Zuber
Note: do not reply to this e-mail address but to the one in my signature below.
--
Prof. Beno�t Zuber
Institute of Anatomy
University of Bern
Baltzerstrasse 2
3000 Bern 9
Switzerland
benoit.zuber-at-ana.unibe.ch
+41 31 631 84 40

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I think you may be confusing with the physical dimensions of the photosite
on the CCD, and the dimensions of the image projected onto one photosite-
which leads to one pixel in the image.

The camera manufacturer should state the size of the photosites in the
spec sheet, if they don't, you can divide the horizontal or vertical
distance by the number of photosites/pixels in that axis.

It that it sounds like you are using a TEM, so in this case there is an
added complexity: (unless you are using a fancy direct electron capture
CCD) the electrons hit a scintillator, this leads to photons being
emitted, and these are then then either reflected off a mirror into a
lens, or down optic fibres to the CCD. The size of the image created by
the electrons on the scitillator may not be the same size as the photon
image reaching the CCD. Also, the electron image is getting larger as you
lower plane of the scintillator. The camera length will be calculated to
the location of the normal view screen, so if you are using that for you
scaling calulations, you have to take into account how the camera length
will be changed by the location of the scintillator plane (if the
scintintillator is say 10cm below the screen, this will not mean the
camera length is 10cm greater! It will require more thought, and possibly
knowing how hight above the view screen and the camera's scintillator the
nodal point of the projector lens is? But my electron optical knowledge is
lacking, sadly).

I have never worked with diffraction images (I am a biologist), and I
don't know what the exact relationship between camera length and
magnification is, or whether diffraction images really have a
'magnification' as such- I am completely ignorant!

Hope that helps you (even if it makes things more confusing in the short
term!),

Ben

--
Research Support Manager
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271649
{http://www.mrc.ox.ac.uk/}

On 14/02/2013 09:33, "amit.welcomes.u-at-gmail.com"
{amit.welcomes.u-at-gmail.com} wrote:

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16, 31 -- From ben.micklem-at-pharm.ox.ac.uk Thu Feb 14 05:33:29 2013
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16, 31 -- From: Ben Micklem {ben.micklem-at-pharm.ox.ac.uk}
16, 31 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
16, 31 -- Subject: Re: [Microscopy] CCD camera sensor size
16, 31 -- Thread-Topic: [Microscopy] CCD camera sensor size
16, 31 -- Thread-Index: AQHOCpaPEiJPCxEQEkaj96oJEVDSvZh5OQcA
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Kristen -

Project MICRO will be delighted to help you as much as we can.
You're already working with MICRO's website, so I'll avoid
duplicating its contents; I'll restrict myself to your specific
concerns.

Teacher's manuals. "Microscopic Explorations" was produced as part of
the Lawrence Hall of Science's Great Explorations in Math and Science
(GEMS) series, with sponsorship by MSA; it's middle school. "Private
Eye" is an independently produced K-12 manual with a different
approach. "Nanoscale Science" is a NSTA manual for grades 6-12.
You'll find full information on the first two in MICRO' main
booklist; "Nanoscale Science" is in the 'Nanotechnology for Kids"
list. A dozen articles about various aspects of microscopy outreach
have appeared recently in MSA's 'Microscopy Today' magazine. You
can read them online; they're listed in the introduction to the MICRO
booklist.

Online lesson. Lesley Bechtold {Lesley.Bechtold-at-jax.org} of the
Jackson Lab in Bar Harbor, Maine has written an online microscopy
lesson that MICRO hopes to offer soon; you might be able to use it.
Contact Lesley directly for information.

Teacher education. Both GEMS and Private Eye offer excellent onsite
teacher training programs.

Microscopes. You say "stereo". That just means "two eyepieces". Do
you mean low power inspection/dissection scopes, compound scopes, or
both? I've got a lot to say about buying school microscopes on the
website, and none of it encourages binocular optics. Monocular is
much cheaper. Rough use is inevitable, and repairing misaligned
optics requires professional help. An estimated 17% of younger
children have some sort of binocular vision problem, and young faces
haven't the eye spacing to use many binocs.

Having said that, my basic advice is that you should work with a
company that specializes in student microscopes, rather than a
general school supply or lab equipment firm. All of the scopes are
Chinese, and they're all quite similar; the difference will be in
quality control and repair service. If you'd like help with onsite
evaluation of samples, MICRO can find an experienced MSA member in
your area.

I welcome your further questions!

Caroline

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Email: kalennon-at-hagerstowncc.edu
Name: Kristen Lennon

Organization: Hagerstown Community College

Title-Subject: [Filtered] microscopy outreach - microscopes

Message: Hello Everyone,

I've just been given the exciting and daunting challenge of creating a
K-12 outreach and teacher education program centered on microscopy.I've
visited the Project Micro web site and am using those resources, but I'm
wondering if any of you have any other suggested resources. I'd also
like some suggestions as to where to buy good quality stereo microscopes
at a K-12 non-profit kind of price. The idea is to get more kids excited
about science through the engaging microscopic world and make microscopy
more accessible to K-12 students and their teachers.

Your advice is, as always, much appreciated.
Best,
Kristen Lennon
kalennon-at-hagerstowncc.edu

--
Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
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There are a few things I am curious about:

Concerning a recent statement.."A monocular microscope is much cheaper.", I started thinking (dangerous!).

Being a user, not a microscope optics technologist, I always had several questions:

1)Don't Binocular microscopes simply split the image? What exactly is the advantage(s)in a compound turret-type microscope)? Isn't this an obsolete function and design?(Pre-electronic camera)?

2)Also, it would seem 'logical' to use ccd cameras and monocular microscopes.
a)since there are fewer optical components in monocular, so less optical distortions(minimal absorption, refraction, diffraction,..?, right?
b) and 'how much' information is lost or degraded by electronic amplification, scanning or dithering(?) artifacts and CCD noise(& etc.)? Are the losses reduced by higher ccd pixel resolution? I have some confusion about CCDs, and I would guess that I am not alone. I suppose the higher number of pixels advertised or stated in CCD specs. is actually suggesting a higher density per square inch..is this equivalent or similar to 'spatial resolution'(and how is it comparable)?

3)And finally, which produces more usable information: monocular, binocular or trinocular microscopes(these more advanced levels would seem to require many more optical components).

(I think this is just the tip of the iceberg..is there a very thorough website for these details..seems like I remember someone mentioning a few...frequently)
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The "recent statement" was mine, in response to a question about
school microscopes. I'll respond to your questions from that point
of view.
1) Some dissecting scopes have 2 fully separate light paths,
providing a true stereo image.; the cheaper ones do not. Such scopes
are used to look at the surface of a sample, so stereo is useful.
Compound scopes are used with transmitted light to view the internal
structure of thin samples. Obsolete? Perhaps eventually, but at the
K-12 level, cost is still in control.
2a) The 2 light paths are parallel rather than in series, so the
defects aren't additive.
2b) Not relevant for basic school microscopes.
3) Silly question. The 3rd tube is just a place to mount a camera
or other device.
The website that you remember is Molecular Expressions; it's very
good. http://micro.magnet.fsu.edu/primer/

Caroline

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Caroline Schooley
Project MICRO Coordinator
Microscopy Society of America
45301 Caspar Point Road, Box 117
Caspar, CA 95420
Phone/FAX (707)964-9460
Project MICRO: http://www.microscopy.org/education/projectMICRO

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Sorry for late reply everyone. thanks for the responses!
-at-Ben- yes you are right i want the physical size of the photosite
which makes one pixel on image. as from it i can actually count
movement in cm etc by measuring pixcels.

Sadly its not a Gatan camera. And procuring a standard sample like
mag*i*cal will not be feasible, vis a vis money and time.

Regards
Amit Gupta

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Maybe I am missing something here. Correct me if I am. Back in the old days, I knew the size of my Polaroid film exactly. That would be like knowing the size of your sensor (or its pixels) exactly. Your question is about the actual magnification.

Do not trust the magnification display on your scope. I have heard that +/- 5% errors are not uncommon. Most systems are better. You need to calibrate the scope under the same conditions that you are using for your sample.

I understand that one of the benefits of Mag*i*cal is that it provides several functions and scales on the same sample so that it is useful at multiple magnifications.

However, the standard may not have to be exotic. You might use a TEM grid and count the grid openings. You can then determine your pixel spacing from the number of pixels across a feature of know dimension. You should also be able to determine the magnification. Just don't assume that the displayed magnification is correct.

Warren S.

-----Original Message-----
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Sorry for late reply everyone. thanks for the responses!
-at-Ben- yes you are right i want the physical size of the photosite
which makes one pixel on image. as from it i can actually count
movement in cm etc by measuring pixcels.

Sadly its not a Gatan camera. And procuring a standard sample like
mag*i*cal will not be feasible, vis a vis money and time.

Regards
Amit Gupta

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The sensor is a part of the optical / detection system. As an image is the output of the instrument, the sensor size creates confusion in the image calibration and can be excluded from calculations:
http://confocal-manawatu.pbworks.com/w/page/60817289/Image%20Calibration

Just try to avoid calibration of your equipment and images by pixel size or scale bar.

Cheers,
Dmitry
Advanced Knowledge Management
for MICROSCOPY and Image Analysis
_________________________________________
Dmitry Sokolov, Ph.D.
Mobile: +64 21 063 5382
dmitry.v.sokolov-at-gmail.com

On 19/02/2013 3:48 a.m., wesaia-at-iastate.edu wrote:
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} Maybe I am missing something here. Correct me if I am. Back in the old days, I knew the size of my Polaroid film exactly. That would be like knowing the size of your sensor (or its pixels) exactly. Your question is about the actual magnification.
}
} Do not trust the magnification display on your scope. I have heard that +/- 5% errors are not uncommon. Most systems are better. You need to calibrate the scope under the same conditions that you are using for your sample.
}
} I understand that one of the benefits of Mag*i*cal is that it provides several functions and scales on the same sample so that it is useful at multiple magnifications.
}
} However, the standard may not have to be exotic. You might use a TEM grid and count the grid openings. You can then determine your pixel spacing from the number of pixels across a feature of know dimension. You should also be able to determine the magnification. Just don't assume that the displayed magnification is correct.
}
} Warren S.
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} Sorry for late reply everyone. thanks for the responses!
} -at-Ben- yes you are right i want the physical size of the photosite
} which makes one pixel on image. as from it i can actually count
} movement in cm etc by measuring pixcels.
}
} Sadly its not a Gatan camera. And procuring a standard sample like
} mag*i*cal will not be feasible, vis a vis money and time.
}
}
} Regards
} Amit Gupta
}
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Hello Amit,

I have been following your posts, and I am not sure I understand why you
need the actual pixel size. If you want to measure diffraction spots or
their position, all you need to do is to insert a specimen with a known
crystal lattice (for example a piece of Si), and calibrate your system for
your camera length. The rest is then just a bit of arithmetic. And the
centers of the diffraction spots should be as accurate as you can get.

The same is true if you want to measure distances in real space. Insert a
calibration sample (for example the ubiquitous line grating), and measure
the distance between lines and use a simple program that allows you to get
the pixel coordinates, and you can calculate a calibration in microns/pixel.
And then you can calculate distances or movements.

Any microscopy software should be able to do this.

Am I missing something?

Can you tell us who the manufacturer of your camera is, or what the model
is? Perhaps there is a way to find the sensor that was used in the camera.

Mike

Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
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Sorry for late reply everyone. thanks for the responses!
-at-Ben- yes you are right i want the physical size of the photosite which
makes one pixel on image. as from it i can actually count movement in cm etc
by measuring pixcels.

Sadly its not a Gatan camera. And procuring a standard sample like mag*i*cal
will not be feasible, vis a vis money and time.

Regards
Amit Gupta

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-at- Warren S.
exactly! I need to calibrate my whole TEM. From magnification to
camera length to aberration constants. for that i need what is the
size of my "polaroid film". since we have a ccd camera instead of
film, I need to know the actual size of ccd sensor (will I not?)

-at- rest (sorry for not addressing individually)
I can caliberate at high mag with gold lattice image, at low mag with
copper grid etc, its the range 80000 to 200000 which is giving bit
trouble.

camera: Olympus Keen View G2

Gatan generally gives a caliberation sample with CCD, alas! olympus dont.

With Regards
Amit

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Hi Amit,

the trick with the microscopes is that they can be considered as a
single "filter" between your sample and your computer screen / eye.

Calibrate the instrument against known (better if with the standard
reference) sample at the magnification and other critical parameters
fixed. The subsequent calibration of the image is a breeze:
Image Size (um) = Maximum Visible Length of Reference (um) * Image
Size (pixels) / Maximum Visible Length of Reference (pixels)

The pixel size at given (same as at the calibration of the instrument!)
imaging conditions is calculated as:
Image Size (um) / Image Size (pixels).

Estimation of the camera pixel size:
Camera pixel size (um) = Maximum Visible Length of Reference (um) /
Maximum Visible Length of Reference (pixels) / Magnification

Here we come to the need of calibration of the microscope in terms of
magnification at the plane of the camera chip that may be tricky. I
would rely in this case on either chip data from the manufacturer of the
camera or direct measurements of the chip size under the binocular
microscope. Don't forget about the LM calibration too. ;0)

This all is described at MIAWiki Knowledge Network:
http://confocal-manawatu.pbworks.com/w/page/60817289/Image%20Calibration
If something is missing or unclear, your comments/suggestions at the
bottom of the page will be highly appreciated.

Please Skype me: FalconDot if I can be of more help in real time.

Cheers,
Dmitry

Advanced Knowledge Management
for MICROSCOPY and Image Analysis
________________________________
Dmitry Sokolov, Ph.D.
Mob: +64 21 063 5382
dmitry.v.sokolov-at-gmail.com

19.02.2013 18:05, amit.welcomes.u-at-gmail.com ��:
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}
} -at- Warren S.
} exactly! I need to calibrate my whole TEM. From magnification to
} camera length to aberration constants. for that i need what is the
} size of my "polaroid film". since we have a ccd camera instead of
} film, I need to know the actual size of ccd sensor (will I not?)
}
} -at- rest (sorry for not addressing individually)
} I can caliberate at high mag with gold lattice image, at low mag with
} copper grid etc, its the range 80000 to 200000 which is giving bit
} trouble.
}
} camera: Olympus Keen View G2
}
} Gatan generally gives a caliberation sample with CCD, alas! olympus dont.
}
} With Regards
} Amit
}
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Amit,

For magnifications where you cannot directly use a standard to calibrate, you can use ratios.

So, for example, starting from the lowest high-magnification where you can calibrate directly from a lattice image, find a region on the sample with two clearly visible and widely separated features - measure the spacing. Drop the magnification and re-measure the spacing. The ratio of the two measurements gives you the ratio of the lower, uncalibrated magnification to the higher, calibrated magnification.

Larry Stoter
(Working on a Microsoft-free computer)

On 19 Feb 2013, at 05:04, amit.welcomes.u-at-gmail.com wrote:

}
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} ----------------------------------------------------------------------------
}
} -at- Warren S.
} exactly! I need to calibrate my whole TEM. From magnification to
} camera length to aberration constants. for that i need what is the
} size of my "polaroid film". since we have a ccd camera instead of
} film, I need to know the actual size of ccd sensor (will I not?)
}
} -at- rest (sorry for not addressing individually)
} I can caliberate at high mag with gold lattice image, at low mag with
} copper grid etc, its the range 80000 to 200000 which is giving bit
} trouble.
}
} camera: Olympus Keen View G2
}
} Gatan generally gives a caliberation sample with CCD, alas! olympus dont.
}
} With Regards
} Amit
}
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Hello Amit,

Can you give me the serial number of the camera? I can then get you the
pixel size of the camera.

For a magnification of 8000 - 20,000 the Diffraction Grating Replica,
available from a number of vendors, should give you what you need to
calibrate your system.

If you look on the DVD or CD for the software that came with your camera,
you should find a "step-by-step" document (in the doc folder), which
explains the calibration routine for the cameras. It is pretty straight
forward. You can store a number of calibration values for different
magnifications, and the software (iTEM) will interpolate between the
calibrated values. Let me know if you found the document. I can also send it
to you, but I do need to know what software and what release you are using.

Can you tell me where the TEM is located?

Mike

Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-234-9270
f: +1-303-202-6350
Mike.Bode-at-ResAltaTech.com
www.ResAltaTech.com

-----Original Message-----
X-from: amit.welcomes.u-at-gmail.com [mailto:amit.welcomes.u-at-gmail.com]
Sent: Monday, February 18, 2013 10:03 PM
To: mike.bode-at-resaltatech.com

-at- Warren S.
exactly! I need to calibrate my whole TEM. From magnification to camera
length to aberration constants. for that i need what is the size of my
"polaroid film". since we have a ccd camera instead of film, I need to know
the actual size of ccd sensor (will I not?)

-at- rest (sorry for not addressing individually) I can caliberate at high mag
with gold lattice image, at low mag with copper grid etc, its the range
80000 to 200000 which is giving bit trouble.

camera: Olympus Keen View G2

Gatan generally gives a caliberation sample with CCD, alas! olympus dont.

With Regards
Amit

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Dear All,

Does anyone have any step-by-step notes they're willing to share on how to
use the strain mapping software within ESVision (including screen shots?)?

I have some handwritten notes taken a few years ago, but they seem to make
little sense with the version of ESVision I am using (Version 5.0).
Likewise, the Help menu on ESVision is hopeless- there is no mention of it
anywhere and certainly nothing on how to set it up.

I'd be interested to hear other recommendations on getting strain maps out
of Emispec files.

Yours, Jon

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Hello listers, got an odd question that I am hoping the more experienced
microanalytical folks can help me with.

I think we are seeing some type of "Escape" peak using an SDD XEDS
system, but these are not the 1.740kev Silicon escape peaks I am used to.
They are much closer to the primary peak, vary in energy displacement from
the primary peaks, and seem to be proportionally larger than Silicon escape
peaks from Si\Li detectors. Examples: 0.372 kev below Ti Ka (-at- 4.508kev),
0.610kev below Ni Ka (-at- 7.471 kev), and 0.646 Kev Cu Ka (-at- 8.040kev).

They are not "system peaks" as they track with the larger primary peaks as
we change samples. I suspect that they are "escape artifacts" of SDD�s and
obviously I do not spend enough time reading "MicroNews" and the
microanalysis literature to be aware of them. Anyone want to help us out,
please?

We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps (TEM
remember?), but have tested with same results at higher 10 - 60kcps.

The Bruker software does not provide any identification markers. As they
are a significant size, if they are an escape artifact of SDD�s, their absence
from the primary peak will significantly effect the quant calculations. Is this
correct?

Thank you in advance!

Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.cami.muohio.edu

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I wonder if it might be some sort of incomplete charge collection or else a calibration error. We ran into incomplete charge collection on a broken Ge detector. It broke after it warmed up and we got a shadowing of the spectrum downscale from where it was suppose to be. As I recall, we found peaks at about 60% of the energy of the main peaks. Yours seems to be consistently around 92% of the energy of the main peak (i.e., ~8% downscale).

I mention calibration error in case different portions of the detector feed into different preamps. (Does anyone do that?) Maybe one segment is out of calibration.

I hope your system is under warranty. I would guess you need to get Bruker in to evaluate the detector and/or its setup. Maybe it is just a calibration issue.

I would be interested in seeing some of the data if you could provide a screen shot or an MSA copy of a spectrum.

Warren

-----Original Message-----
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To: wesaia-at-iastate.edu

Hello listers, got an odd question that I am hoping the more experienced
microanalytical folks can help me with.

I think we are seeing some type of "Escape" peak using an SDD XEDS
system, but these are not the 1.740kev Silicon escape peaks I am used to.
They are much closer to the primary peak, vary in energy displacement from
the primary peaks, and seem to be proportionally larger than Silicon escape
peaks from Si\Li detectors. Examples: 0.372 kev below Ti Ka (-at- 4.508kev),
0.610kev below Ni Ka (-at- 7.471 kev), and 0.646 Kev Cu Ka (-at- 8.040kev).

They are not "system peaks" as they track with the larger primary peaks as
we change samples. I suspect that they are "escape artifacts" of SDD�s and
obviously I do not spend enough time reading "MicroNews" and the
microanalysis literature to be aware of them. Anyone want to help us out,
please?

We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps (TEM
remember?), but have tested with same results at higher 10 - 60kcps.

The Bruker software does not provide any identification markers. As they
are a significant size, if they are an escape artifact of SDD�s, their absence
from the primary peak will significantly effect the quant calculations. Is this
correct?

Thank you in advance!

Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.cami.muohio.edu

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O.k., well thank you folks for thinking about it.

Here is an example of what we are seeing:

http://www.cami.muohio.edu/xeds/Weirdpeak1.jpg

Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.cami.muohio.edu

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Warren:

Not a calibration error, as the peaks above and below all align where they
should. Hmm, but I hope its not a "shadowing as you suggest.

Image at:

http://www.cami.muohio.edu/xeds/Weirdpeak1.jpg

On 21 Feb 2013 at 17:18, Straszheim, Warren E [BIOTC] wrote:

} I wonder if it might be some sort of incomplete charge collection or
} else a calibration error. We ran into incomplete charge collection on
} a broken Ge detector. It broke after it warmed up and we got a
} shadowing of the spectrum downscale from where it was suppose to be.
} As I recall, we found peaks at about 60% of the energy of the main
} peaks. Yours seems to be consistently around 92% of the energy of the
} main peak (i.e., ~8% downscale).
}
} I mention calibration error in case different portions of the detector
} feed into different preamps. (Does anyone do that?) Maybe one segment
} is out of calibration.
}
} I hope your system is under warranty. I would guess you need to get
} Bruker in to evaluate the detector and/or its setup. Maybe it is just
} a calibration issue.
}
} I would be interested in seeing some of the data if you could provide
} a screen shot or an MSA copy of a spectrum.
}
} Warren
}
} -----Original Message-----
} From: Edelmare-at-miamioh.edu [mailto:Edelmare-at-miamioh.edu]
} Sent: Thursday, February 21, 2013 10:45 AM
} To: wesaia-at-iastate.edu
} Subject: [Microscopy] Subject: Unidentified extra XEDS peaks
}
}
}
}
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} ------
}
}
} Hello listers, got an odd question that I am hoping the more
} experienced microanalytical folks can help me with.
}
} I think we are seeing some type of "Escape" peak using an SDD XEDS
} system, but these are not the 1.740kev Silicon escape peaks I am used
} to. They are much closer to the primary peak, vary in energy
} displacement from the primary peaks, and seem to be proportionally
} larger than Silicon escape peaks from Si\Li detectors. Examples:
} 0.372 kev below Ti Ka (-at- 4.508kev), 0.610kev below Ni Ka (-at- 7.471
} kev), and 0.646 Kev Cu Ka (-at- 8.040kev).
}
} They are not "system peaks" as they track with the larger primary
} peaks as we change samples. I suspect that they are "escape
} artifacts" of SDD�s and obviously I do not spend enough time reading
} "MicroNews" and the microanalysis literature to be aware of them.
} Anyone want to help us out, please?
}
} We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
} 200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps
} (TEM remember?), but have tested with same results at higher 10 -
} 60kcps.
}
} The Bruker software does not provide any identification markers. As
} they are a significant size, if they are an escape artifact of SDD�s,
} their absence from the primary peak will significantly effect the
} quant calculations. Is this correct?
}
}
} Thank you in advance!
}
}
} Richard E. Edelmann, Ph.D., Director
} Center for Advanced Microscopy & Imaging
} 9C Upham Hall
} Miami University, Oxford, OH 45056
} Ph: 513.529.5712 Fax: 513.529.4243
} E-mail: edelmare-at-muohio.edu
} http://www.cami.muohio.edu
}
}
}
} ==============================Original
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Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.cami.muohio.edu

==============================Original Headers==============================
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I am by no means very knowledgeable in the subject, but isn't that a sum
peak? ( Ti Ka1+2)

Regards,
Darrell

Edelmare-at-miamioh.edu wrote on 02/21/2013 11:44:39 AM:

} From: Edelmare-at-miamioh.edu
} To: Darrell Miles/Fishkill/IBM-at-IBMUS,
} Date: 02/21/2013 11:55 AM
} Subject: [Microscopy] Subject: Unidentified extra XEDS peaks
}
}
}
}
}
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}
} Hello listers, got an odd question that I am hoping the more experienced

} microanalytical folks can help me with.
}
} I think we are seeing some type of "Escape" peak using an SDD XEDS
} system, but these are not the 1.740kev Silicon escape peaks I am used
to.
} They are much closer to the primary peak, vary in energy displacement
from
} the primary peaks, and seem to be proportionally larger than Silicon
escape
} peaks from Si\Li detectors. Examples: 0.372 kev below Ti Ka (-at-
4.508kev),
} 0.610kev below Ni Ka (-at- 7.471 kev), and 0.646 Kev Cu Ka (-at- 8.040kev).
}
} They are not "system peaks" as they track with the larger primary peaks
as
} we change samples. I suspect that they are "escape artifacts" of SDD�s
and
} obviously I do not spend enough time reading "MicroNews" and the
} microanalysis literature to be aware of them. Anyone want to help us
out,
} please?
}
} We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
} 200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps (TEM
} remember?), but have tested with same results at higher 10 - 60kcps.
}
} The Bruker software does not provide any identification markers. As
they
} are a significant size, if they are an escape artifact of SDD�s,
} their absence
} from the primary peak will significantly effect the quant
} calculations. Is this
} correct?
}
}
} Thank you in advance!
}
}
} Richard E. Edelmann, Ph.D., Director
} Center for Advanced Microscopy & Imaging
} 9C Upham Hall
} Miami University, Oxford, OH 45056
} Ph: 513.529.5712 Fax: 513.529.4243
} E-mail: edelmare-at-muohio.edu
} http://www.cami.muohio.edu
}
}
}
} ==============================Original
Headers==============================
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Actually, Warren's idea is compelling, even though I have not heard of this before. The energy ratio of the "shadow" peak to the characteristic peak is constant at 0.918+-0.01. Sounds like there are two calibrations acting here with different gains.
Ken

On Feb 21, 2013, at 1:04 PM, Edelmare-at-miamioh.edu wrote:

}
}
}
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} Warren:
}
} Not a calibration error, as the peaks above and below all align where they
} should. Hmm, but I hope its not a "shadowing as you suggest.
}
} Image at:
}
} http://www.cami.muohio.edu/xeds/Weirdpeak1.jpg
}
}
}
}
} On 21 Feb 2013 at 17:18, Straszheim, Warren E [BIOTC] wrote:
}
} } I wonder if it might be some sort of incomplete charge collection or
} } else a calibration error. We ran into incomplete charge collection on
} } a broken Ge detector. It broke after it warmed up and we got a
} } shadowing of the spectrum downscale from where it was suppose to be.
} } As I recall, we found peaks at about 60% of the energy of the main
} } peaks. Yours seems to be consistently around 92% of the energy of the
} } main peak (i.e., ~8% downscale).
} }
} } I mention calibration error in case different portions of the detector
} } feed into different preamps. (Does anyone do that?) Maybe one segment
} } is out of calibration.
} }
} } I hope your system is under warranty. I would guess you need to get
} } Bruker in to evaluate the detector and/or its setup. Maybe it is just
} } a calibration issue.
} }
} } I would be interested in seeing some of the data if you could provide
} } a screen shot or an MSA copy of a spectrum.
} }
} } Warren
} }
} } -----Original Message-----
} } From: Edelmare-at-miamioh.edu [mailto:Edelmare-at-miamioh.edu]
} } Sent: Thursday, February 21, 2013 10:45 AM
} } To: wesaia-at-iastate.edu
} } Subject: [Microscopy] Subject: Unidentified extra XEDS peaks
} }
} }
} }
} }
} } ----------------------------------------------------------------------
} } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
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} }
} }
} } Hello listers, got an odd question that I am hoping the more
} } experienced microanalytical folks can help me with.
} }
} } I think we are seeing some type of "Escape" peak using an SDD XEDS
} } system, but these are not the 1.740kev Silicon escape peaks I am used
} } to. They are much closer to the primary peak, vary in energy
} } displacement from the primary peaks, and seem to be proportionally
} } larger than Silicon escape peaks from Si\Li detectors. Examples:
} } 0.372 kev below Ti Ka (-at- 4.508kev), 0.610kev below Ni Ka (-at- 7.471
} } kev), and 0.646 Kev Cu Ka (-at- 8.040kev).
} }
} } They are not "system peaks" as they track with the larger primary
} } peaks as we change samples. I suspect that they are "escape
} } artifacts" of SDD�s and obviously I do not spend enough time reading
} } "MicroNews" and the microanalysis literature to be aware of them.
} } Anyone want to help us out, please?
} }
} } We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
} } 200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps
} } (TEM remember?), but have tested with same results at higher 10 -
} } 60kcps.
} }
} } The Bruker software does not provide any identification markers. As
} } they are a significant size, if they are an escape artifact of SDD�s,
} } their absence from the primary peak will significantly effect the
} } quant calculations. Is this correct?
} }
} }
} } Thank you in advance!
} }
} }
} } Richard E. Edelmann, Ph.D., Director
} } Center for Advanced Microscopy & Imaging
} } 9C Upham Hall
} } Miami University, Oxford, OH 45056
} } Ph: 513.529.5712 Fax: 513.529.4243
} } E-mail: edelmare-at-muohio.edu
} } http://www.cami.muohio.edu
} }
} }
} }
} } ==============================Original
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} Richard E. Edelmann, Ph.D., Director
} Center for Advanced Microscopy & Imaging
} 9C Upham Hall
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|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
Director, The High-Resolution Analytical Electron Microbeam Facility
of the Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| | | :|

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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Like I said, I'm not very knowledgeable. :-D

I just showed how daft I am, and how long it has been since I looked at
this stuff. I had looked at the image, and missed that it is on the wrong
side to be a sum. I am interested in what the real answer is. Just for
fun, I got out my old Tracor slide chart and PGT X-Ray Energy "ruler". I
poked around a bit, but nothing makes sense, unless you have Scandium in
your sample. So, I will wait for someone who knows what they are talking
about, and learn...

Regards,
Darrell

"Straszheim, Warren E [BIOTC]" {wesaia-at-iastate.edu} wrote on 02/21/2013
03:44:46 PM:

} From: "Straszheim, Warren E [BIOTC]" {wesaia-at-iastate.edu}
} To: Darrell Miles/Fishkill/IBM-at-IBMUS,
} Date: 02/21/2013 03:47 PM
} Subject: RE: [Microscopy] Re: Subject: Unidentified extra XEDS peaks
}
} Did you check the link to the JPG file that he offered? There are
} peaks that are downscale and proportionate to the major peaks.
}
} I see sum peaks often in our Oxford system when I am looking at Al
} and push the dead time up toward 30%. I keep finding "argon" at 3 keV.
}
} -----Original Message-----
} From: milesd-at-us.ibm.com [mailto:milesd-at-us.ibm.com]
} Sent: Thursday, February 21, 2013 2:41 PM
} To: wesaia-at-iastate.edu
} Subject: [Microscopy] Re: Subject: Unidentified extra XEDS peaks
}
}
}
}
}
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} I am by no means very knowledgeable in the subject, but isn't that a sum

} peak? ( Ti Ka1+2)
}
} Regards,
} Darrell
}
}
} Edelmare-at-miamioh.edu wrote on 02/21/2013 11:44:39 AM:
}
} } From: Edelmare-at-miamioh.edu
} } To: Darrell Miles/Fishkill/IBM-at-IBMUS,
} } Date: 02/21/2013 11:55 AM
} } Subject: [Microscopy] Subject: Unidentified extra XEDS peaks
} }
} }
} }
} }
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} }
} } Hello listers, got an odd question that I am hoping the more
experienced
}
} } microanalytical folks can help me with.
} }
} } I think we are seeing some type of "Escape" peak using an SDD XEDS
} } system, but these are not the 1.740kev Silicon escape peaks I am used
} to.
} } They are much closer to the primary peak, vary in energy displacement
} from
} } the primary peaks, and seem to be proportionally larger than Silicon
} escape
} } peaks from Si\Li detectors. Examples: 0.372 kev below Ti Ka (-at-
} 4.508kev),
} } 0.610kev below Ni Ka (-at- 7.471 kev), and 0.646 Kev Cu Ka (-at- 8.040kev).
} }
} } They are not "system peaks" as they track with the larger primary
peaks
} as
} } we change samples. I suspect that they are "escape artifacts" of
SDD�s
} and
} } obviously I do not spend enough time reading "MicroNews" and the
} } microanalysis literature to be aware of them. Anyone want to help us
} out,
} } please?
} }
} } We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
} } 200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps
(TEM
} } remember?), but have tested with same results at higher 10 - 60kcps.
} }
} } The Bruker software does not provide any identification markers. As
} they
} } are a significant size, if they are an escape artifact of SDD�s,
} } their absence
} } from the primary peak will significantly effect the quant
} } calculations. Is this
} } correct?
} }
} }
} } Thank you in advance!
} }
} }
} } Richard E. Edelmann, Ph.D., Director
} } Center for Advanced Microscopy & Imaging
} } 9C Upham Hall
} } Miami University, Oxford, OH 45056
} } Ph: 513.529.5712 Fax: 513.529.4243
} } E-mail: edelmare-at-muohio.edu
} } http://www.cami.muohio.edu
} }
} }
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear all,

I'm 99.9% sure this is Scandium as well. The Gaussian peakshape and position matches Sc-Ka whereby the Sc-Kb is hidden under the Ti-Ka.

Some comments on the shadowing phenomena on SDD: We do have 3 SDD in house which do have shadow peaks on the left side of the main peak but this is only clearly noticable in the energy region below 1 keV. In this region, the resolution gets worse, the main peak reduces in height and a satelite peak shows up on the left side of the main peak. The peakposition of the main peak is also moving to a lower energy. Therefore analysis of B, C, N, O, and F and/or working at low kV is getting more and more problematic over time.

According to the SDD crystal supplier, X-ray photons can permanently damage the crystal structure resulting in the phenomena described. The guaranteed number of counts hitting the detector without causing damaging is 1.0E+12. This number seems to be enormous, but the reality is, using a detector at 60.000 counts/sec, 24 hrs/day, 7 days/wk, it can be used in an optimal condition for only half a year. Even just doing imaging on the microscope is contributing to the crystal damage (reported on the microscopy listserver in oct 2011).

Regards,

Corrie

________________________________________
X-from: milesd-at-us.ibm.com [milesd-at-us.ibm.com]
Sent: Thursday, February 21, 2013 23:41
To: Hoek, Corrie van

Like I said, I'm not very knowledgeable. :-D

I just showed how daft I am, and how long it has been since I looked at
this stuff. I had looked at the image, and missed that it is on the wrong
side to be a sum. I am interested in what the real answer is. Just for
fun, I got out my old Tracor slide chart and PGT X-Ray Energy "ruler". I
poked around a bit, but nothing makes sense, unless you have Scandium in
your sample. So, I will wait for someone who knows what they are talking
about, and learn...

Regards,
Darrell

"Straszheim, Warren E [BIOTC]" {wesaia-at-iastate.edu} wrote on 02/21/2013
03:44:46 PM:

} From: "Straszheim, Warren E [BIOTC]" {wesaia-at-iastate.edu}
} To: Darrell Miles/Fishkill/IBM-at-IBMUS,
} Date: 02/21/2013 03:47 PM
} Subject: RE: [Microscopy] Re: Subject: Unidentified extra XEDS peaks
}
} Did you check the link to the JPG file that he offered? There are
} peaks that are downscale and proportionate to the major peaks.
}
} I see sum peaks often in our Oxford system when I am looking at Al
} and push the dead time up toward 30%. I keep finding "argon" at 3 keV.
}
} -----Original Message-----
} From: milesd-at-us.ibm.com [mailto:milesd-at-us.ibm.com]
} Sent: Thursday, February 21, 2013 2:41 PM
} To: wesaia-at-iastate.edu
} Subject: [Microscopy] Re: Subject: Unidentified extra XEDS peaks
}
}
}
}
}
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}
} I am by no means very knowledgeable in the subject, but isn't that a sum

} peak? ( Ti Ka1+2)
}
} Regards,
} Darrell
}
}
} Edelmare-at-miamioh.edu wrote on 02/21/2013 11:44:39 AM:
}
} } From: Edelmare-at-miamioh.edu
} } To: Darrell Miles/Fishkill/IBM-at-IBMUS,
} } Date: 02/21/2013 11:55 AM
} } Subject: [Microscopy] Subject: Unidentified extra XEDS peaks
} }
} }
} }
} }
} }
}
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} }
} }
} } Hello listers, got an odd question that I am hoping the more
experienced
}
} } microanalytical folks can help me with.
} }
} } I think we are seeing some type of "Escape" peak using an SDD XEDS
} } system, but these are not the 1.740kev Silicon escape peaks I am used
} to.
} } They are much closer to the primary peak, vary in energy displacement
} from
} } the primary peaks, and seem to be proportionally larger than Silicon
} escape
} } peaks from Si\Li detectors. Examples: 0.372 kev below Ti Ka (-at-
} 4.508kev),
} } 0.610kev below Ni Ka (-at- 7.471 kev), and 0.646 Kev Cu Ka (-at- 8.040kev).
} }
} } They are not "system peaks" as they track with the larger primary
peaks
} as
} } we change samples. I suspect that they are "escape artifacts" of
SDD�s
} and
} } obviously I do not spend enough time reading "MicroNews" and the
} } microanalysis literature to be aware of them. Anyone want to help us
} out,
} } please?
} }
} } We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
} } 200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps
(TEM
} } remember?), but have tested with same results at higher 10 - 60kcps.
} }
} } The Bruker software does not provide any identification markers. As
} they
} } are a significant size, if they are an escape artifact of SDD�s,
} } their absence
} } from the primary peak will significantly effect the quant
} } calculations. Is this
} } correct?
} }
} }
} } Thank you in advance!
} }
} }
} } Richard E. Edelmann, Ph.D., Director
} } Center for Advanced Microscopy & Imaging
} } 9C Upham Hall
} } Miami University, Oxford, OH 45056
} } Ph: 513.529.5712 Fax: 513.529.4243
} } E-mail: edelmare-at-muohio.edu
} } http://www.cami.muohio.edu
} }
} }
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I vote for the "new artifact from new SDD" as explained by Ritchie et al
in their 2011 Microscopy and Microanalysis article (vol 17, pp 903-910):
Compton Scattering Artifacts in Electron Excited X-Ray Spectra Measured
with a Silicon Drift Detector
by Nicholas W.M. Ritchie,* Dale E. Newbury, and Abigail P. Lindstrom

(I am using it in my electron microprobe class this semester, so have a
link to it, if you need it...
{www.geology.wisc.edu/~johnf/g777/777MMarticles2.html}

John Fournelle

On 2/21/13 9:48 AM, Edelmare-at-miamioh.edu wrote:
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}
} Hello listers, got an odd question that I am hoping the more experienced
} microanalytical folks can help me with.
}
} I think we are seeing some type of "Escape" peak using an SDD XEDS
} system, but these are not the 1.740kev Silicon escape peaks I am used to.
} They are much closer to the primary peak, vary in energy displacement from
} the primary peaks, and seem to be proportionally larger than Silicon escape
} peaks from Si\Li detectors. Examples: 0.372 kev below Ti Ka (-at- 4.508kev),
} 0.610kev below Ni Ka (-at- 7.471 kev), and 0.646 Kev Cu Ka (-at- 8.040kev).
}
} They are not "system peaks" as they track with the larger primary peaks as
} we change samples. I suspect that they are "escape artifacts" of SDD�s and
} obviously I do not spend enough time reading "MicroNews" and the
} microanalysis literature to be aware of them. Anyone want to help us out,
} please?
}
} We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
} 200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps (TEM
} remember?), but have tested with same results at higher 10 - 60kcps.
}
} The Bruker software does not provide any identification markers. As they
} are a significant size, if they are an escape artifact of SDD�s, their absence
} from the primary peak will significantly effect the quant calculations. Is this
} correct?
}
}
} Thank you in advance!
}
}
} Richard E. Edelmann, Ph.D., Director
} Center for Advanced Microscopy & Imaging
} 9C Upham Hall
} Miami University, Oxford, OH 45056
} Ph: 513.529.5712 Fax: 513.529.4243
} E-mail: edelmare-at-muohio.edu
} http://www.cami.muohio.edu
}
}
}
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--
John Fournelle
Senior Scientist
Director, Electron Probe and SEM Lab
University of Wisconsin Dept of Geoscience
Madison, Wisconsin 53706
Cell 608-438-7480

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} realname - alaa afeef
} Email - alaa.afeef-at-gmail.com
} ORGANIZATION - Glasgow University
} EDUCATION - Graduate College
} LOCATION - Glasgow
} SUBJECT_OF_QUESTION - regular sample with geometrical shape
} QUESTION -
} May I have your help with the below issue:
} I am trying to look for a sample of a 50-200 nm size to do Electron Tomography (tilted tomo),
}
} Actually, I am trying to find something that is already characterized, with geometrical shape and a well known morphology.
}
} I would be very grateful for your help.
}
} Wish you a nice weekend!
} Ala'

==============================Original Headers==============================
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I'd try carbon black, an ASTM N500 grade. It has the morphology and size you're interested in and depending on your definition of characterized- it's been well studied.

Failing that, I try kaolinite clay. Hexagonal shaped plates, xtal system known, crystallographic and chemical properties well documented.

My two cents..........

Stay safe.............
Frank

} May I have your help with the below issue:
} I am trying to look for a sample of a 50-200 nm size to do Electron Tomography (tilted tomo),
}
} Actually, I am trying to find something that is already characterized, with geometrical shape and a well known morphology.
}
} I would be very grateful for your help.
}
} Wish you a nice weekend!
} Ala'

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Here is a question for Richard...
When you said "Examples: 0.372 kev below Ti Ka (-at- 4.508kev), 0.610kev below Ni Ka (-at- 7.471 kev), and 0.646 Kev Cu Ka (-at- 8.040kev)." These were examples from different samples, each having a Ni or Cu as the major element?

Question for John Fournelle...
Can you give more specifics about the new artifact? I'm interested.
Ken

On Feb 21, 2013, at 11:47 AM, Edelmare-at-miamioh.edu wrote:

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}
} Hello listers, got an odd question that I am hoping the more experienced
} microanalytical folks can help me with.
}
} I think we are seeing some type of "Escape" peak using an SDD XEDS
} system, but these are not the 1.740kev Silicon escape peaks I am used to.
} They are much closer to the primary peak, vary in energy displacement from
} the primary peaks, and seem to be proportionally larger than Silicon escape
} peaks from Si\Li detectors. Examples: 0.372 kev below Ti Ka (-at- 4.508kev),
} 0.610kev below Ni Ka (-at- 7.471 kev), and 0.646 Kev Cu Ka (-at- 8.040kev).
}
} They are not "system peaks" as they track with the larger primary peaks as
} we change samples. I suspect that they are "escape artifacts" of SDD�s and
} obviously I do not spend enough time reading "MicroNews" and the
} microanalysis literature to be aware of them. Anyone want to help us out,
} please?
}
} We are working with a JEOL-2100, LaB6 at 200kV, and a Bruker Quantax
} 200 SDD. Using a beryllium holder. Count rates of 0.7 - 2.0 Kcps (TEM
} remember?), but have tested with same results at higher 10 - 60kcps.
}
} The Bruker software does not provide any identification markers. As they
} are a significant size, if they are an escape artifact of SDD�s, their absence
} from the primary peak will significantly effect the quant calculations. Is this
} correct?
}
}
} Thank you in advance!
}
}
} Richard E. Edelmann, Ph.D., Director
} Center for Advanced Microscopy & Imaging
} 9C Upham Hall
} Miami University, Oxford, OH 45056
} Ph: 513.529.5712 Fax: 513.529.4243
} E-mail: edelmare-at-muohio.edu
} http://www.cami.muohio.edu
}
}
}
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|_|"|_|"|_|_|"|_|"|_|"|_|_|"|_|"|_|_|"|_|"|_|"|_|_|
Kenneth JT Livi, PhD
Director, The High-Resolution Analytical Electron Microbeam Facility
of the Integrated Imaging Center
Departments of Earth and Planetary Sciences and Biology
Olin Hall
3400 N Charles Street
Johns Hopkins University
Baltimore, Maryland 21218 USA
| | | .| | .| | .| | .| | | :| | | .| | .| | .| | .| | | :|

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To the listserver,

Some time ago was a posted query regarding embedding large tissues in LR White. As an alternative to flat embedding molds, I suggest using polyethylene Wheaton Snap Caps for LR White embedding. They are available from Fisher in 22mm diameter (06-450-201) and also in larger diameters. To drive out inherent moisture in the caps, store them in the oven prior to use. Once the sample is embedded in LR White within the caps, place these directly in a 60C oven for polymerization, without any cover, with the media directly exposed to the atmosphere within the oven. Importantly, together with the samples, place either a 500ml beaker of dry ice or a small, uncovered thermos of liquid nitrogen. The sublimation of CO2 or evaporation of nitrogen displaces enough of the atmosphere within the oven cavity for quality polymerization of the media. There may be a thin gooey layer at the top of the caps but this can be wiped clean with ethanol. The media separates easily from the caps. Including an Aclar film in the bottom of the cap works well for a crystal-clear view of the embedded sample.

Doug

Douglas R. Keene
Assistant Investigator
Micro-Imaging Center
Shriners Hospital for Children
3101 SW Sam Jackson Park Road
Portland, Oregon 97239

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Hello Everyone,
Out of left field comes: Does anyone run an aggregate particle sizing on precipitated silica by TEM? We measure the primary particle size by TEM, but we normally have to breakup the aggregate in silica recovered from rubber by ashing. Is there an ASTM or other procedure anyone can recommend?

Thanks!!!

Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Hi Dmitry,

Even that nice explanation is still simplified. Nowadays, microscopes have many different modes and many different detectors, hence even a single microscope system has to be considered and many 'filters'. Depending on the mode of operation, and if it has been previously accurately aligned and calibrated, these factors will determine if the actual pixel size stored with the data is actually correct for the microscope, the kV, and the mode of operation used.

FEI has software that tries to semi-automate the calibration routine, which can be stored for any camera, any mode and any kV. However, it still requires the user to recall the file that is the correct calibration for the operational conditions that are applicable. i.e. a camera at the end of an imaging filter may be calibrated for TEM mode, however if EFTEM mode is engaged to improve the usability of this detector with a reduction in effective magnification of about 10X, the user would like the calibration to be updated automatically too.

Many camera manufacturers assume that once a calibration is done and valid for a camera this is the complete picture, furthermore usually the camera SW allows any operator to change the calibrations without verifying if 'administrator' or 'supervisor' credentials are in place, and therefore all is typically NOT well; and what is good today is not necessarily right tomorrow!

In plain language this allows any operator to 'mess' with the calibrations. Nothing can be trusted. If the camera manufacturers remain in charge of the calibrations in such a manner, it is only a matter of time before wrong calibrations are used for data analysis and interpretation which is what currently happens very frequently.

Different modes:-
LM mode
LM STEM mode
TEM mode (35mm ccd camera)
TEM mode (bottom-mount camera)
EFTEM mode (specialized projection lens series to be used in TEM mode with energy filters) Diffraction mode EFTEM Diffraction mode (specialized projection lens series to be used in diffraction mode with an energy filter) STEM mode EFTSTEM mode a small camera length series to be used for optimized STEM/EELS experiments.

And, to complete the necessary scenario's, all combinations of modes with different kV's.

Keeping systems calibrated and producing accurately calibrated images is much more complex than initially thought. Even if all calibrations are done accurately; If the sample height at the time of imaging/analysis is not correct, all of the calibrations can be undone by varying the primary focusing lens too far from the 'calibrated' values.

The FEI MagCal software goes some way towards having a properly calibrated data acquisition system by allowing a semi-automated calibration routine for all different modes and all different detectors, however users still have to be vigilant to ensure the system is correctly calibrated for the kV and mode currently selected by recalling the appropriate file. However once this file is locked in by an administrator, at least the calibrations cannot be altered within the file.

Sincerely, Jan

P.S. Disclaimer: I work for FEI, manufacturer of Electron Microscope Systems, who also develop and produce SW to aid users to correctly calibrate and quantify data obtained with electron microscope systems.

-----Original Message-----
X-from: dmitry.v.sokolov-at-gmail.com [mailto:dmitry.v.sokolov-at-gmail.com]
Sent: Tuesday, February 19, 2013 1:34 AM
To: Ringnalda, Jan

Hi Amit,

the trick with the microscopes is that they can be considered as a single "filter" between your sample and your computer screen / eye.

Calibrate the instrument against known (better if with the standard
reference) sample at the magnification and other critical parameters fixed. The subsequent calibration of the image is a breeze:
Image Size (um) = Maximum Visible Length of Reference (um) * Image Size (pixels) / Maximum Visible Length of Reference (pixels)

The pixel size at given (same as at the calibration of the instrument!) imaging conditions is calculated as:
Image Size (um) / Image Size (pixels).

Estimation of the camera pixel size:
Camera pixel size (um) = Maximum Visible Length of Reference (um) / Maximum Visible Length of Reference (pixels) / Magnification

Here we come to the need of calibration of the microscope in terms of magnification at the plane of the camera chip that may be tricky. I would rely in this case on either chip data from the manufacturer of the camera or direct measurements of the chip size under the binocular microscope. Don't forget about the LM calibration too. ;0)

This all is described at MIAWiki Knowledge Network:
http://confocal-manawatu.pbworks.com/w/page/60817289/Image%20Calibration
If something is missing or unclear, your comments/suggestions at the bottom of the page will be highly appreciated.

Please Skype me: FalconDot if I can be of more help in real time.

Cheers,
Dmitry

Advanced Knowledge Management
for MICROSCOPY and Image Analysis
________________________________
Dmitry Sokolov, Ph.D.
Mob: +64 21 063 5382
dmitry.v.sokolov-at-gmail.com

19.02.2013 18:05, amit.welcomes.u-at-gmail.com ��:
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}
} -at- Warren S.
} exactly! I need to calibrate my whole TEM. From magnification to
} camera length to aberration constants. for that i need what is the
} size of my "polaroid film". since we have a ccd camera instead of
} film, I need to know the actual size of ccd sensor (will I not?)
}
} -at- rest (sorry for not addressing individually) I can caliberate at
} high mag with gold lattice image, at low mag with copper grid etc, its
} the range 80000 to 200000 which is giving bit trouble.
}
} camera: Olympus Keen View G2
}
} Gatan generally gives a caliberation sample with CCD, alas! olympus dont.
}
} With Regards
} Amit
}
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Thank you Jan, that's a good point!

What you suggest below for the TEM calibration procedure is rewritten in
the Knowledge Network format and published in MIAWiki.

TEM Magnification Factors:
http://confocal-manawatu.pbworks.com/w/page/63920300/TEM%20Magnification%20Factors

TEM Modes:
http://confocal-manawatu.pbworks.com/w/page/63920396/TEM%20Modes

MagCal:
http://confocal-manawatu.pbworks.com/w/page/63920440/MagCal

TEM Calibration Best Practices:
http://confocal-manawatu.pbworks.com/w/page/63920519/TEM%20Calibration%20Best%20Practices

Your corrections, suggestions and comments would be highly appreciated.

Regarding the simplification, the degree of abstraction/details depends
on the problem to be solve. Too many details make the decision making
difficult. That is the same kind of phenomena as with the signal/noise
ratio in microscopy. I believe if users remember to have the calibration
done/applied at all the critical parameters fixed, that will do the job.
Your detailed comments however put the focus further into how namely
should the calibration be done. Many thanks for that again.

With kind regards,
Dmitry

Advanced Knowledge Management
for MICROSCOPY and Image Analysis
________________________________
Dmitry Sokolov, Ph.D.
Mob: +64 21 063 5382
dmitry.v.sokolov-at-gmail.com

23.02.2013 11:35, Jan.Ringnalda-at-fei.com ��:
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} Hi Dmitry,
}
} Even that nice explanation is still simplified. Nowadays, microscopes have many different modes and many different detectors, hence even a single microscope system has to be considered and many 'filters'. Depending on the mode of operation, and if it has been previously accurately aligned and calibrated, these factors will determine if the actual pixel size stored with the data is actually correct for the microscope, the kV, and the mode of operation used.
}
} FEI has software that tries to semi-automate the calibration routine, which can be stored for any camera, any mode and any kV. However, it still requires the user to recall the file that is the correct calibration for the operational conditions that are applicable. i.e. a camera at the end of an imaging filter may be calibrated for TEM mode, however if EFTEM mode is engaged to improve the usability of this detector with a reduction in effective magnification of about 10X, the user would like the calibration to be updated automatically too.
}
} Many camera manufacturers assume that once a calibration is done and valid for a camera this is the complete picture, furthermore usually the camera SW allows any operator to change the calibrations without verifying if 'administrator' or 'supervisor' credentials are in place, and therefore all is typically NOT well; and what is good today is not necessarily right tomorrow!
}
} In plain language this allows any operator to 'mess' with the calibrations. Nothing can be trusted. If the camera manufacturers remain in charge of the calibrations in such a manner, it is only a matter of time before wrong calibrations are used for data analysis and interpretation which is what currently happens very frequently.
}
} Different modes:-
} LM mode
} LM STEM mode
} TEM mode (35mm ccd camera)
} TEM mode (bottom-mount camera)
} EFTEM mode (specialized projection lens series to be used in TEM mode with energy filters) Diffraction mode EFTEM Diffraction mode (specialized projection lens series to be used in diffraction mode with an energy filter) STEM mode EFTSTEM mode a small camera length series to be used for optimized STEM/EELS experiments.
}
} And, to complete the necessary scenario's, all combinations of modes with different kV's.
}
} Keeping systems calibrated and producing accurately calibrated images is much more complex than initially thought. Even if all calibrations are done accurately; If the sample height at the time of imaging/analysis is not correct, all of the calibrations can be undone by varying the primary focusing lens too far from the 'calibrated' values.
}
} The FEI MagCal software goes some way towards having a properly calibrated data acquisition system by allowing a semi-automated calibration routine for all different modes and all different detectors, however users still have to be vigilant to ensure the system is correctly calibrated for the kV and mode currently selected by recalling the appropriate file. However once this file is locked in by an administrator, at least the calibrations cannot be altered within the file.
}
} Sincerely, Jan
}
} P.S. Disclaimer: I work for FEI, manufacturer of Electron Microscope Systems, who also develop and produce SW to aid users to correctly calibrate and quantify data obtained with electron microscope systems.
}
}
} -----Original Message-----
} X-from: dmitry.v.sokolov-at-gmail.com [mailto:dmitry.v.sokolov-at-gmail.com]
} Sent: Tuesday, February 19, 2013 1:34 AM
} To: Ringnalda, Jan
} Subject: [Microscopy] Re: CCD sensor size
}
}
}
}
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} Hi Amit,
}
} the trick with the microscopes is that they can be considered as a single "filter" between your sample and your computer screen / eye.
}
} Calibrate the instrument against known (better if with the standard
} reference) sample at the magnification and other critical parameters fixed. The subsequent calibration of the image is a breeze:
} Image Size (um) = Maximum Visible Length of Reference (um) * Image Size (pixels) / Maximum Visible Length of Reference (pixels)
}
} The pixel size at given (same as at the calibration of the instrument!) imaging conditions is calculated as:
} Image Size (um) / Image Size (pixels).
}
} Estimation of the camera pixel size:
} Camera pixel size (um) = Maximum Visible Length of Reference (um) / Maximum Visible Length of Reference (pixels) / Magnification
}
} Here we come to the need of calibration of the microscope in terms of magnification at the plane of the camera chip that may be tricky. I would rely in this case on either chip data from the manufacturer of the camera or direct measurements of the chip size under the binocular microscope. Don't forget about the LM calibration too. ;0)
}
} This all is described at MIAWiki Knowledge Network:
} http://confocal-manawatu.pbworks.com/w/page/60817289/Image%20Calibration
} If something is missing or unclear, your comments/suggestions at the bottom of the page will be highly appreciated.
}
} Please Skype me: FalconDot if I can be of more help in real time.
}
} Cheers,
} Dmitry
}
} Advanced Knowledge Management
} for MICROSCOPY and Image Analysis
} ________________________________
} Dmitry Sokolov, Ph.D.
} Mob: +64 21 063 5382
} dmitry.v.sokolov-at-gmail.com
}
} 19.02.2013 18:05, amit.welcomes.u-at-gmail.com ��:
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} }
} } -at- Warren S.
} } exactly! I need to calibrate my whole TEM. From magnification to
} } camera length to aberration constants. for that i need what is the
} } size of my "polaroid film". since we have a ccd camera instead of
} } film, I need to know the actual size of ccd sensor (will I not?)
} }
} } -at- rest (sorry for not addressing individually) I can caliberate at
} } high mag with gold lattice image, at low mag with copper grid etc, its
} } the range 80000 to 200000 which is giving bit trouble.
} }
} } camera: Olympus Keen View G2
} }
} } Gatan generally gives a caliberation sample with CCD, alas! olympus dont.
} }
} } With Regards
} } Amit
} }
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Email: akomakov-at-physics.siu.edu
Name: Andrei Kolmakov

Organization: SIUC

Title-Subject: [Filtered] FEG Hitachi 4500

Message: Dear Colleague,
We are using refurbished Hitachi 4500 FEG SEM for about two years. The instrument is out of service
and I don't know when last time it was serviced. Lately new symptoms start to develop: the emission
current is constantly going down and multiple fleshing does not help the current does not recover
or stabilize. The preset value can only be reached for a short time only after HV turned Off/ON. Any
advice. If it is FEG dying does somebody have a manual to replace, adjust it?
Thank you for your comments in advance.
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Organization: Beckman Res. Inst. City of Hope

Title-Subject: [Filtered] Stereomicroscope System for LEICA EM AFS2

Message: Dear Colleagues,

We need a stereomicroscope for the LEICA freeze substitution system so as to be able to see
particular types of specimens. Our impression is that sometime these steromicroscopes are purchased
but not needed or used. Before ordering a new one we thought it might be good to ask if anyone have
has one that is not needed and that we could be purchase perhaps at a savings compared to the
current price of a new one. Below is the part number and description from the Leica website.

Always looking to make our limited dollars stretch as far as is reasonable. Best wishes, Marcia Miller

16907105 Stereomicroscope System for EM AFS2
consisting of:
-Stereo carrier
-S6E Stereomicroscope
-0.5 x Objective
-eyepieces 10 x (2 pcs.)

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Hi Ala',
MgO is very good for this - lots of very small perfect cubes. Note that there are hazards involved in the specimen prep protocol below and you MUST do a proper risk assessment. Perhaps one of the few TEM specimen preparations where temporary/permanent blindness is a real possibility!

You will need:
A couple of cm of Mg ribbon
Some tongs
Some welding goggles or similar; better with a very dark glass sheet you can put in front of the burning metal
A flame source (e.g. Bunsen burner)
A fume hood
Some carbon-coated TEM grids
Tweezers
Also it can be easier with someone else to help.

Set up the bunsen burner in the fume hood. One person holds the Mg ribbon in the tongs; the other takes a TEM grid in the tweezers. Alternatively hold the grid in reverse-action tweezers in a clamp. The aim is to set the Mg alight and waft the TEM grid in the smoke. The difficulty is that Mg burns with a very intense white light that you must not look at directly - you must put the goggles on before setting the metal alight. This of course makes it difficult to see anything else and why it can be easier with an extra pair of helping hands.

With a little practice this is not too difficult to do. MgO is moisture-sensitive so the samples are best put in the TEM vacuum within an hour or so - the cubes will decompose somewhat after a day.

Good luck and stay safe

Richard
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} realname - alaa afeef
} Email - alaa.afeef-at-gmail.com
} ORGANIZATION - Glasgow University
} EDUCATION - Graduate College
} LOCATION - Glasgow
} SUBJECT_OF_QUESTION - regular sample with geometrical shape
} QUESTION -
} May I have your help with the below issue:
} I am trying to look for a sample of a 50-200 nm size to do Electron Tomography (tilted tomo),
}
} Actually, I am trying to find something that is already characterized, with geometrical shape and a well known morphology.
}
} I would be very grateful for your help.
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Hello listers, well we found our unidentified peaks. They were caused by our
over working of our XEDS system. Thanks to the good folks at Bruker.
Shutting the system down and allowing it to cool for a hour then restarting
cleared the "extra peaks".

With respects to Dr. Ritchie et al. Like with any new technology new
"quirks" (aka "Features") will arise, and it takes experience to learn the pit
falls to watch out for, and once again the microscopy list comes through in
letting us all learn a little more.

The one thing I do wish to make clear, as we all should realize that in
naming specific vendors in postings in this list, it is done so that the specifics
of a saturation are known, and NEVER should be taken as an off-handed
criticism of a vendor. I for one very much appreciate vendors who attend
and post responses to the list. I think we all benefit from open discussions.

Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
http://www.cami.muohio.edu

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Dear Listers,

In fixing brain tissue, is one type of buffer better than any other. Here are the conditions, perfusion fixation, 2.5% glutaraldehyde + 2% paraformaldehyde, pH 7.2 to 7.4. Buffers in question are .1m Sodium Phosphate buffer vs. .1M Sodium Cacodylate buffer. Is one better than the other? Also could there be some other buffer that is better? I am not involved in a project involving brain tissue, but I have been asked my opinion. In my experience I have seen neural tissues that have been fixed in fixatives that have used both types of buffers and the fixation in any of those tissues was fine. However to provide the individual with as much advice as possible, I would like to pass on the advice and opinions of others, especially anyone out there who has a lot of experience with neural tissue. As always thanks in advance.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
tbargar-at-unmc.edu
402-559-7347

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Good morning,

Dear Tom, (dear all)
just to keep this reply on ��major issue�� very short�� (for sure no complete or exhaustive answers at all!)

Unfortunately you have not given more detailed information about type and use of tissue after such a perfusion fixation, guessing it will be TEM (and - perhaps ? as a prerequisite for such, LM on semithin sections).

1) IMHO, in the first place the most annoying issue (esp. for BRAIN) isn��t choice of ��buffer�� but instead: parameters of perfusion, like perfusion PRESSURE, volume/min; way of application of perfusion.

2) second place: CAVE: ischemic conditions before and during preparation for brain perfusion.

3) buffers: usually perfusion with fixative is preceded by flushing vasculature with ��Ringer solutions�� to get vessels free from plasma & cells (RBC��s, etc.). Most faults are done within the first mentioned steps.

4) it depends really whether you are using 0.1 M PO4 or 0.1M CAC-buffer�� to the former you cannot add osmolality {active} ions, to the latter you can (CaCl2, MgCl2 etc) without forcing precipitation. For example, Millonig's 0.13M or other, (brain-compatible) molarity will be one out of suitable PO4-buffers (I personally would not use commercially available brands of PO4-buffers!)
Decision also to use saccharose or glucose as an additive, also usually for 3) not few methods in the past have been using {Tyrode solution} (there are several recipes for this), or other mammalian ��Ringer�� solutions. Also there are a lot of methods using polyvinylpyrrolidone (e.g. PVP-40, to adjust the correct colloid osmotic pressure) ) �� try Google Search for: e.g. { polyvinylpyrrolidone & perfusion fixation of Brain } ...yes, I see: 71.900 results! but perhaps the first two results-pages will give you an overview...

5) Choice of Perfusion technique, contents of flushing buffer/Ringer solutions as well as fixatives (Pre-fix, ��main-fix��, post-fix) depending also a little bit on the conception what will be done afterwards: LM (semithin sections and e.g. 3D-(serial) reconstruction), HC-IHC, etc.), EM, etc.

So - to be short enough ? it also will depend on the animal which��s brain (the brain of which?) is to be perfused.
Some recipes / techniques {in stock} , needing a little bit more of information on the project.

Best wishes and regards
Wolfgang MUSS, PhD
Member of MSA
EM-Lab
SALK-LKH (Gen. Hospital) SALZBURG
AUSTRIA
PS: sorry for poor English, but it is to early...

Reminder for MAY 2013:
+++ 40th Annual SCUR (Society for Cutaneous Ultrastructure Research) Meeting +++
SALZBURG, Austria
6th Joint Meeting with the SSSR (Society for Skin Structure Research, Japan)
Sunday, 12th until Tuesday, 14th May 2013
Call for papers, Registration open
All Information needed:
cf. website at: http://orgs.dermis.net/scur/scur2013/content/e01home/e01home/index_ger.html

Von: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu]
Gesendet: Montag, 25. Februar 2013 23:47
An: Mu�� Wolfgang
Betreff: [Microscopy] choice of buffer in fixation of neural tissue

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Dear Listers,

In fixing brain tissue, is one type of buffer better than any other.
Here are the conditions, perfusion fixation, 2.5% glutaraldehyde + 2% paraformaldehyde, pH 7.2 to 7.4. Buffers in question are 0.1M Sodium Phosphate buffer vs. 0.1M Sodium Cacodylate buffer. Is one better than the other? Also could there be some other buffer that is better? I am not involved in a project involving brain tissue, but I have been asked my opinion.
In my experience I have seen neural tissues that have been fixed in fixatives that have used both types of buffers and the fixation in any of those tissues was fine.
However to provide the individual with as much advice as possible, I would like to pass on the advice and opinions of others, especially anyone out there who has a lot of experience with neural tissue. As always thanks in advance.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
tbargar-at-unmc.edu
402-559-7347

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Good morning, Tom,

I have been doing neural work for 6 years now on a focused basis, but E.M. for 39 years. Phosphate buffer is my buffer of choice for all tissue, because it does a better job of preserving tissue cytosol than cacodylate. The only problem with phosphate that you can run into is that you have to keep it fresh, and to rinse it out of tissue well, or it will precipitate in your tissue, giving you a fine pepper in your tissue. If you use cacodylate, you will get beautiful tissue, but you will loose some of the ground substance in your cells, the cells will be less dense than with phosphate buffer. It depends, then, on what you are looking for. You will almost never run into precipitate problems with cacodylate buffer. The buffer is toxic, having arsenic in it, so it's not as good for our environment, and is more of a hassle to dispose of with your Health and Safety people. Phosphate will give you a more life-like presentation of tissue, preserving more of the proteins and ground substance in your cells, but if it's mishandled, you will get precipitates. Uranyl acetate will react with phosphate buffer, forming uranyl phosphate, which is insoluble, for instance, so you need to watch your en-bloc staining step, if you do this proceedure. For our neural work, we've been perfusing with 4% E.M. grade paraformaldehyde with 4% sucrose, allowing fixation overnight or over the weekend, doing the dissection, then fixing with glutaraldehyde. We get some tissue tearing upon dissection, but using this method, we can submit some of our tissue for histology and some for E.M. It's something for your associates to consider.

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
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Sent: Monday, February 25, 2013 5:50 PM
To: Haller, Edward

Dear Listers,

In fixing brain tissue, is one type of buffer better than any other. Here are the conditions, perfusion fixation, 2.5% glutaraldehyde + 2% paraformaldehyde, pH 7.2 to 7.4. Buffers in question are .1m Sodium Phosphate buffer vs. .1M Sodium Cacodylate buffer. Is one better than the other? Also could there be some other buffer that is better? I am not involved in a project involving brain tissue, but I have been asked my opinion. In my experience I have seen neural tissues that have been fixed in fixatives that have used both types of buffers and the fixation in any of those tissues was fine. However to provide the individual with as much advice as possible, I would like to pass on the advice and opinions of others, especially anyone out there who has a lot of experience with neural tissue. As always thanks in advance.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
tbargar-at-unmc.edu
402-559-7347

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Email: amit.welcomes.u-at-gmail.com
Name: Amit

Organization: IISc

Title-Subject: [Filtered] CCD camera sensor size

Message: Sorry everyone for disappearing, was trying to calibrate the microscope.
First of all thank you for all the suggestions and patience.
Though even after 8th attempt I am still not able to replicate value of spherical abberation
constant, but we will not yield!!!!

-at-Xing- i dont know about 2010, but 2100 that we are using has an option of "Lens relaxation" an i
believe it must be there in 2010 also, though its location might have been buried under menus. I am
rying to calibrate by keeping objective lens at "std focus" and trying to focus using Z-controls.

-at-Warren- problem lies in that 1) we do not have proper std, 2) still we will need "actual distance"
as in film. thats why i am trying to find the sensor size.

-at- Mike Bode- thanks for your interest, as far geographical location is concerned it is in Bangalore,
India. I tried looking on the camera but serial number was not there. Unfortunately i dont have
access to its installation certificate etc, so i am not sure if i will be able to provide you with
its serial number. Still i will be grateful if you can just provide the same for the latest release
that you can. iTEM is version 5.2.Thank again

-at-Larry Stoter- Thats a nice idea! Thanks!

-at-Ray Twesten- Thanks for going through the pain of taking and pasting snapshots.

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Email: krpeters-at-mtu.edu
Name: Karl Peterson

Organization: U of Toronto

Title-Subject: [Filtered] welding tungsten filaments to posts

Message: Dear DIYers out there:

We are working with an x-ray CT machine that burns through filaments on a weekly basis; not from
mis-use, it's just the nature of the beast. We are looking into the feasibility of spot welding our
own tungsten filaments to the posts that stick out of the ceramic block inside the electron gun.
We've located a business that will sell us filaments of the correct diameter, and bent in the same
hair-pin shape as what we are currently using. We've also found on campus a lab with a tiny
spot-welder, and they're willing to let us use it.

Having never tried anything of the sort, I have some questions:

1) The business that sells the filaments has two grades: lamp-grade tungsten, and a "3RW" alloy made
with 3% Re. They said the Re-bearing alloy lasts longer. I figure we'll try it. Is there any reason
not to?(Using SEM/EDS I couldn't tell whether or not there was trace rhenium in our old filaments,
as the peaks would be right next to tungsten's, and buried in the shoulder)

2) The lab with the spot welder said they just used some "general" electrode tips, and thought they
should work OK, but that it might depend on the composition of the posts. The posts are some kind of
tungsten iron chromium alloy. Does this raise any warning flags? (I don't want to wreck their welder)

3) I am accustomed to cleaning the gun, and replacing/centering filaments for an SEM, but have
always ordered filaments that came already pre-attached to the ceramic block. I couldn't find
anything with the right dimensions from standard SEM suppliers. So, we plan to
re-use the old blocks, but they have a noticeable build-up of I'm guessing mostly evaporated
tungsten. Should I worry about cleaning this stuff off?

Any advice people might have about making their own filaments would be greatly appreciated. As an
undergrad, I had a resourceful professor who made his own filaments, which gave me the idea. I also
remember him sticking a paperclip (in place of whatever was supposed be there) in a busted Pirani
vacuum gage. Come to think of it, he also came to lecture once with giant bandages on his hand, from
trying to pull something out of a running lawn-mower.

Thanks,
kp
--
Karl Peterson, Univ. of Toronto, Dept. of Civil Eng.
35 Saint George Street, Toronto, ON M5S 1A4, CANADA
phone 1.416.978.4589 fax 1.416.978.6813

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Name: Rick Thompson

Organization: Bmi Surplus

Title-Subject: [Filtered] Is this an ion testing chamber?

Message: I'm trying to find out if this (URL below) is a an ion testing chamber.
Does anyone have an idea?

http://blog.bmius.com/

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I training in college (UICC) on a micro-probe and the filaments were welded to the electrode post. This was years ago. They had a full time electro-tech who kept the electronics operating and made filaments. He use to tell the students he'd kill anyone who burnt out a filament because they were so difficult to weld. I'm sure that was hyperbola, but the police did find bodies in the tennis courts overnight my first two quarters.

I'd like to hear more of your progress.

Frank

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Email: krpeters-at-mtu.edu
Name: Karl Peterson

Organization: U of Toronto

Title-Subject: [Filtered] welding tungsten filaments to posts

Message: Dear DIYers out there:

We are working with an x-ray CT machine that burns through filaments on a weekly basis; not from
mis-use, it's just the nature of the beast. We are looking into the feasibility of spot welding our
own tungsten filaments to the posts that stick out of the ceramic block inside the electron gun.
We've located a business that will sell us filaments of the correct diameter, and bent in the same
hair-pin shape as what we are currently using. We've also found on campus a lab with a tiny
spot-welder, and they're willing to let us use it.

Having never tried anything of the sort, I have some questions:

1) The business that sells the filaments has two grades: lamp-grade tungsten, and a "3RW" alloy made
with 3% Re. They said the Re-bearing alloy lasts longer. I figure we'll try it. Is there any reason
not to?(Using SEM/EDS I couldn't tell whether or not there was trace rhenium in our old filaments,
as the peaks would be right next to tungsten's, and buried in the shoulder)

2) The lab with the spot welder said they just used some "general" electrode tips, and thought they
should work OK, but that it might depend on the composition of the posts. The posts are some kind of
tungsten iron chromium alloy. Does this raise any warning flags? (I don't want to wreck their welder)

3) I am accustomed to cleaning the gun, and replacing/centering filaments for an SEM, but have
always ordered filaments that came already pre-attached to the ceramic block. I couldn't find
anything with the right dimensions from standard SEM suppliers. So, we plan to
re-use the old blocks, but they have a noticeable build-up of I'm guessing mostly evaporated
tungsten. Should I worry about cleaning this stuff off?

Any advice people might have about making their own filaments would be greatly appreciated. As an
undergrad, I had a resourceful professor who made his own filaments, which gave me the idea. I also
remember him sticking a paperclip (in place of whatever was supposed be there) in a busted Pirani
vacuum gage. Come to think of it, he also came to lecture once with giant bandages on his hand, from
trying to pull something out of a running lawn-mower.

Thanks,
kp
--
Karl Peterson, Univ. of Toronto, Dept. of Civil Eng.
35 Saint George Street, Toronto, ON M5S 1A4, CANADA
phone 1.416.978.4589 fax 1.416.978.6813

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Hello All,

I'm hoping to find out some information for our Zeiss LEO 1550 SEM, specifically in repairing/replacing our column isolation valve or the bellows within it. We have an issue with our SEM when the valve opens between the gun and the chamber that we lose significant amounts of vacuum. Being a schottky gun, this is not ideal.

If anyone has any experiences with dealing with cracks or other failures of the column isolation valves, possibly with the bellows, could you please share them? Or if there is a known third party supplier of Zeiss LEO 1550 parts, possibly second hand, we would appreciate hearing about them.

Much appreciation,

Andrew
Valued Quality. Delivered.
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http://www.intertek.com

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Title-Subject: [Filtered] ImmunoGold Summer Workshop

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Karl:

We fabricated our own filaments for the electron microscopes in our
student lab for many years. It is not particularly difficult to do,
once you work out the system, and the results are very satisfactory -
and it saves quite a bit of money.

You should have no trouble spot welding tungsten filaments to the
posts on your filament disks. Mostly, it takes a steady hand and
some care to get the filaments properly centered. Most spot welders
have copper rods for contacts, because they have high electrical and
thermal conductivities and therefore don't stick to most metals in
the welding process. You will need to experiment a bit to get the
proper time and current values to get a good weld without burning
through the wire. The tungsten alloy containing Re will probably
give you a bit longer life, and I see no reason for not using it. It
probably is not absolutely necessary to clean the deposit off the
ceramic base, but you can probably do it easily with a mildly
alkaline solution, maybe Alcanox, or sodium carbonate, or dilute
sodium hydroxide.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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Dear colleagues,

I would like to inform you about a job opening for an electron
microscopy facility head at the Campus Science Support Facilities in
Vienna, Austria.

The CSF Electron Microscopy Facility offers a very wide range of
preparation and visualization techniques for biological samples from
various sources. The technologies on offer comprise conventional and
cryo-methods for sample preparation together with TEM (Morgagni,
Tecnai 20), cryo-TEM (Polara), tomography, EELS, and SEM for data
acquisition.

We are looking for candidates (m/f) with

* PhD in Biology, Biochemistry, Molecular Biology, or a related field

* Several years of research experience; highly skilled in conventional
and cryo-TEM of biological specimens

* Solid technical understanding of electron microscopes and instruments
for sample preparation in biology

* Previous leadership experience, together with project and resource
management skills, will be an asset

The EM Facility Head is expected to be a strategic thinker with strong
organizational, communication, and social skills. Excellent command of
English (spoken and written) is a must, German skills are an asset.

The complete version of the job advertisement can be found at

http://www.csf.ac.at/fileadmin/downloads/open_positions/position_em_head.pdf

Please contact me for questions, applications should be sent to
barbara.miksch-at-csf.ac.at. Review of applications will begin on March
25, 2013.

Best regards,

Guenter

--
Dr. Guenter Resch
Head of Electron Microscopy, Campus Science Support Facilities GmbH
P: +43 (1) 796 23 24 - 7120; F -227120; W: http://www.csf.ac.at/em
Post: Dr. Bohr-Gasse 3, 1030 Vienna, Austria, European Union

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A Research Associate III position is available to be filled immediately in the Electron Microscopy Laboratory at the Frederick National Laboratory for Cancer Research. The Electron Microscopy Laboratory (EML) is a core facility equipped with four TEMs (FEI T20 and T12 and Hitachi H7600 and H7650) and an SEM (Hitachi S-3000). The Facility provides state-of-the-art EM service to the National Cancer Institute.

JOB DESCRIPTION

The Research Associate III will perform biological sample preparation and imaging for traditional transmission electron microscopy (TEM) and scanning electron microscopy (SEM) including room-temperature and cryo ultramicrotomy, as well as immuno-gold TEM and SEM applications. Will also perform image analysis and three dimensional reconstructions on data from negative stained and cryo-electron microscopy and electron tomography. Specific duties include (i) sample preparation for electron microscopy studies (SEM, room temperature plastic embedding, ultramicrotomy, cryo-electron microscopy); (ii) operation of electron microscopes (SEM and TEM) and cryogenic instrumentation; (iii) image analysis and three dimensional reconstructions of electron microscopy data; (iv) segmentation, interpretation and presentation of three-dimensional volume data.

Qualifications:
BASIC QUALIFICATIONS

- Possession of a Bachelors degree from an accredited college/university in a field related to Computational Science, Physics, Biology, Chemistry or four (4) years of related experience in lieu of degree. Foreign degrees must be evaluated for U.S. equivalency.
- In addition to educational requirements, a minimum of eight (8) years related biomedical experience.
- Proficiency in biological and/or material sample preparation for electron microscopy.
- Proficiency in the use and maintenance of transmission electron microscopes.
- Strong understanding of ultrastructure of cells and tissues.
- Ability to function independently and effectively within a team in a specialized research laboratory using originality and sound judgment to perform tasks.
- Experience with Linux/Unix.
- This position is subject to obtaining a Public Trust Clearance.

PREFERRED QUALIFICATIONS

- Proficiency in the use and maintenance of scanning electron microscopes.
- Outstanding computational skills.
- Experience with scripting or other programming skills.
- Experience with computational image analysis with electron microscopy data (e.g. Eman, Spider, Imagic, etc.).
- Experience with cryogenic liquids.
- Experience with maintenance of advanced instrumentation.

Please submit your application online at
http://jobs.saic.com/job/Frederick-Research-Associate-III-321714-%28NCI%29-Job-MD-21701/2417904/

Lauri Rimorin, PHR
Sr. Employment Specialist
SAIC-Frederick, Inc.
Frederick National Laboratory for Cancer Research
Email: rimorinla-at-mail.nih.gov

==============================Original Headers==============================
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Hi Andrew.

Yes, that would be a leaky bellows. I've seen my share of those. Other than contacting Zeiss, http://microscopy.zeiss.com, I'm not sure where you could pick one up.

Andy Zobel
Instrument Technician III

R J Lee Group

www.rjlg.com

724.325.1776 Office
724.325.1776 Direct
724.733.1799 Fax

350 Hochberg Road | Monroeville, PA 15146

Please let us know if we've met your expectations on your project by visiting our Customer Survey.

-----Original Message-----
X-from: andrew.ornelas-at-intertek.com [mailto:andrew.ornelas-at-intertek.com]
Sent: Tuesday, February 26, 2013 5:32 PM
To: Andy Zobel

Hello All,

I'm hoping to find out some information for our Zeiss LEO 1550 SEM, specifically in repairing/replacing our column isolation valve or the bellows within it. We have an issue with our SEM when the valve opens between the gun and the chamber that we lose significant amounts of vacuum. Being a schottky gun, this is not ideal.

If anyone has any experiences with dealing with cracks or other failures of the column isolation valves, possibly with the bellows, could you please share them? Or if there is a known third party supplier of Zeiss LEO 1550 parts, possibly second hand, we would appreciate hearing about them.

Much appreciation,

Andrew
Valued Quality. Delivered.
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http://www.intertek.com

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I have a couple of Gemini columns of ICT version in my storage and would
happily give you CIV bellows, but I can't guarantee wither they will be
compatible with Zeiss/LEO or not - some parts are interchangeable, but
some aren't....

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com

On 2/27/2013 3:24 PM, azobel-at-rjlg.com wrote:
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} Hi Andrew.
}
} Yes, that would be a leaky bellows. I've seen my share of those. Other than contacting Zeiss, http://microscopy.zeiss.com, I'm not sure where you could pick one up.
}
}
} Andy Zobel
} Instrument Technician III
}
} R J Lee Group
}
} www.rjlg.com
}
} 724.325.1776 Office
} 724.325.1776 Direct
} 724.733.1799 Fax
}
} 350 Hochberg Road | Monroeville, PA 15146
}
} Please let us know if we've met your expectations on your project by visiting our Customer Survey.
}
} -----Original Message-----
} X-from: andrew.ornelas-at-intertek.com [mailto:andrew.ornelas-at-intertek.com]
} Sent: Tuesday, February 26, 2013 5:32 PM
} To: Andy Zobel
} Subject: [Microscopy] LEO 1550 Column Isolation Valve
}
}
}
}
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
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}
} Hello All,
}
} I'm hoping to find out some information for our Zeiss LEO 1550 SEM, specifically in repairing/replacing our column isolation valve or the bellows within it. We have an issue with our SEM when the valve opens between the gun and the chamber that we lose significant amounts of vacuum. Being a schottky gun, this is not ideal.
}
} If anyone has any experiences with dealing with cracks or other failures of the column isolation valves, possibly with the bellows, could you please share them? Or if there is a known third party supplier of Zeiss LEO 1550 parts, possibly second hand, we would appreciate hearing about them.
}
} Much appreciation,
}
} Andrew
} Valued Quality. Delivered.
} ------------------------------------------------------------------------------
} CONFIDENTIALITY NOTICE
}
} This email may contain confidential or privileged information, if you are not the intended recipient, or the person responsible for delivering the message to the intended recipient then please notify us by return email immediately. Should you have received this email in error then you should not copy this for any purpose nor disclose its contents to any other person.
}
} http://www.intertek.com
}
}
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} 25, 25 -- Subject: [Microscopy] LEO 1550 Column Isolation Valve
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Karl,
I would be inclined to clean the bases with NaOH just to avoid any bypass
currents. The pins should be of a particular alloy to match the coefficient
of expansion of the particular ceramic being used. As examples, look up
Invar and Kovar. The first has a very low CoE and the second has a CoE that
matches borosilicate glass so it can be used for feedthroughs in vacuum
tubes, hot cathode ion gauges, etc. What ever the alloy is, it probably
won't create any problems for you with the spot welding.

Ken Converse,
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

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Email: krpeters-at-mtu.edu
Name: Karl Peterson

Organization: U of Toronto

Title-Subject: [Filtered] welding tungsten filaments to posts

Message: Dear DIYers out there:

We are working with an x-ray CT machine that burns through filaments on a
weekly basis; not from
mis-use, it's just the nature of the beast. We are looking into the
feasibility of spot welding our
own tungsten filaments to the posts that stick out of the ceramic block
inside the electron gun.
We've located a business that will sell us filaments of the correct
diameter, and bent in the same
hair-pin shape as what we are currently using. We've also found on campus a
lab with a tiny
spot-welder, and they're willing to let us use it.

Having never tried anything of the sort, I have some questions:

1) The business that sells the filaments has two grades: lamp-grade
tungsten, and a "3RW" alloy made
with 3% Re. They said the Re-bearing alloy lasts longer. I figure we'll try
it. Is there any reason
not to?(Using SEM/EDS I couldn't tell whether or not there was trace rhenium
in our old filaments,
as the peaks would be right next to tungsten's, and buried in the shoulder)

2) The lab with the spot welder said they just used some "general" electrode
tips, and thought they
should work OK, but that it might depend on the composition of the posts.
The posts are some kind of
tungsten iron chromium alloy. Does this raise any warning flags? (I don't
want to wreck their welder)

3) I am accustomed to cleaning the gun, and replacing/centering filaments
for an SEM, but have
always ordered filaments that came already pre-attached to the ceramic
block. I couldn't find
anything with the right dimensions from standard SEM suppliers. So, we plan
to
re-use the old blocks, but they have a noticeable build-up of I'm guessing
mostly evaporated
tungsten. Should I worry about cleaning this stuff off?

Any advice people might have about making their own filaments would be
greatly appreciated. As an
undergrad, I had a resourceful professor who made his own filaments, which
gave me the idea. I also
remember him sticking a paperclip (in place of whatever was supposed be
there) in a busted Pirani
vacuum gage. Come to think of it, he also came to lecture once with giant
bandages on his hand, from
trying to pull something out of a running lawn-mower.

Thanks,
kp
--
Karl Peterson, Univ. of Toronto, Dept. of Civil Eng.
35 Saint George Street, Toronto, ON M5S 1A4, CANADA
phone 1.416.978.4589 fax 1.416.978.6813

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Bellows?
I'd try Kurt Lesker Company at WWW.lesker.com.

I had to order several fittings to connect out old vacuum pump to newer filters and components. They were very helpful.

I have no interest in Lesker, I don't even have a current catalog.

Stay safe............
Frank

-----Original Message-----
X-from: azobel-at-rjlg.com [mailto:azobel-at-rjlg.com]
Sent: Wednesday, February 27, 2013 3:32 PM
To: Frank Karl

Hi Andrew.

Yes, that would be a leaky bellows. I've seen my share of those. Other than contacting Zeiss, http://microscopy.zeiss.com, I'm not sure where you could pick one up.

Andy Zobel
Instrument Technician III

R J Lee Group

www.rjlg.com

724.325.1776 Office
724.325.1776 Direct
724.733.1799 Fax

350 Hochberg Road | Monroeville, PA 15146

Please let us know if we've met your expectations on your project by visiting our Customer Survey.

-----Original Message-----
X-from: andrew.ornelas-at-intertek.com [mailto:andrew.ornelas-at-intertek.com]
Sent: Tuesday, February 26, 2013 5:32 PM
To: Andy Zobel

Hello All,

I'm hoping to find out some information for our Zeiss LEO 1550 SEM, specifically in repairing/replacing our column isolation valve or the bellows within it. We have an issue with our SEM when the valve opens between the gun and the chamber that we lose significant amounts of vacuum. Being a schottky gun, this is not ideal.

If anyone has any experiences with dealing with cracks or other failures of the column isolation valves, possibly with the bellows, could you please share them? Or if there is a known third party supplier of Zeiss LEO 1550 parts, possibly second hand, we would appreciate hearing about them.

Much appreciation,

Andrew
Valued Quality. Delivered.
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Earlier today I got into a discussion with a knowledgeable colleague on what the maximum thickness of a plastic resin embedded section should be routinely cut on an ultramicrotome. We both agreed that semi-thin sections in the neighborhood of 0.4 to 0.9 microns was best for high quality LM work. I have always been taught to avoid cutting resin sections thicker than 1.0 micron since it caused undue stress on the microtome's specimen arm.� She said she routinely cuts sections at 2.5 microns.� Am I being overly cautious?� I await your opinions. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

==============================Original Headers==============================
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Hi All

This is admittedly a weird one. I am looking for a commercial lab that
might be able to deposit a thin carbon film on a whole 300mm wafer.

We are having terrible ESD type damages during Circuit Edit.
We have tried a few grounding tricks and they help but we are looking
for a robust solution.

We have had good luck using the Gatan PECS for carbon coating small samples.
The film was thick enuf to stop charging but thin enuf to not short out the device mod.

Thanks very much for your help

bryan tracy
in Sunnyvale

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7, 35 -- CC: "Andreasyan, Armen" {Armen.Andreasyan-at-spansion.com} ,
7, 35 -- "Koo, Heather"
7, 35 -- {Heather.Koo-at-spansion.com} ,
7, 35 -- "Hulog, Jose" {Jose.Hulog-at-spansion.com} ,
7, 35 -- "Vu,
7, 35 -- Joseph" {Joseph.Vu-at-spansion.com} ,
7, 35 -- "Vargas, Mario"
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7, 35 -- "Li, Susan" {Susan.Li-at-spansion.com} , "dl.MCL"
7, 35 -- {dl.MCL-at-spansion.com}
7, 35 -- Subject: Coating 300mm wafers for Circuit Edit
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Ted - Thanks for the insightful comments. I love getting the "source" information on how these things developed. I think we are on the cusp of the generation that has really witnessed the full span of EM being on the verge of retirement and therefore it is great to get the perspective of someone who has seen it all. Best, Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: Ted Pella [mailto:ted_pella-at-tedpella.com]
Sent: Friday, March 01, 2013 10:41 AM
To: Phillips, Thomas E.
Cc: Rick Giberson; Cindy Smith

Earlier today I got into a discussion with a knowledgeable colleague on what
the maximum thickness of a plastic resin embedded section should be
routinely cut on an ultramicrotome. We both agreed that semi-thin sections
in the neighborhood of 0.4 to 0.9 microns was best for high quality LM work.
I have always been taught to avoid cutting resin sections thicker than 1.0
micron since it caused undue stress on the microtome's specimen arm.� She
said she routinely cuts sections at 2.5 microns.� Am I being overly
cautious?� I await your opinions. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

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32, 36 -- To: Ted Pella {ted_pella-at-tedpella.com}
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I am writing this email to urge the TEM (and STEM) communities to
start to take action to reduce misuse of the term "Z-contrast" before
things get too far out of hand, hopefully it is not too late. I am
seeing more and more people use this term when there is clear
strain/diffraction contrast in the images due to the inner collection
angle being too small. I can (will) blame some of the manufacturers
for this, and they certainly need to start teaching their technicians
to get things right.

If we do not as a community act to stop this, the world of non-experts
will make more and more errors in interpreting images.

Please forward this as appropriate to others.

--
Professor Laurence Marks
Department of Materials Science and Engineering
MSE Rm 2036 Cook Hall
2220 N Campus Drive
Northwestern University
Evanston, IL 60208, USA
Tel: (847) 491-3996 Fax: (847) 491-7820
L-marks at northwestern dot edu
www.numis.northwestern.edu

==============================Original Headers==============================
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4, 27 -- Subject: Misuse (abuse) of the term "Z-contrast"
4, 27 -- From: L Marks {marksmsa-at-gmail.com}
4, 27 -- To: microscopy {Microscopy-at-microscopy.com}
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Dear Colleagues,

We would like to make an annoucement on an international conference
"Electron Microscopy and Multiscale Modeling 2013
(November 10th to 14th, 2013, Kyoto, Japan)."

----------------------------------------------------------------
Electron Microscopy and Multiscale Modeling (EMMM) is an
interdisciplinary conference bringing together experts from
several areas of modern electron microscopy, crystallography, and
materials science. It will focus on the link between
methods of interrogating structures at the nano- and meso-scale using
electron microscopy and diffraction,
and the rapidly expanding field of multiscale materials modeling.
EMMM 2013 is a follow-up of the successful EMMM conferences held in
Moscow in 2007, Zurich in 2009, Tahoe (California) in 2011.
----------------------------------------------------------------

Detailed information on the conference will be shown on the following web site:
http://tem-defect.jp/emmm2013/index.html

--
Laurence Marks
Department of Materials Science and Engineering
MSE Rm 2036 Cook Hall
2220 N Campus Drive
Northwestern University
Evanston, IL 60208, USA
Tel: (847) 491-3996 Fax: (847) 491-7820
L-marks at northwestern dot edu
www.numis.northwestern.edu

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***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
**any reply should go directly to the poster**
as well as to the list.
Using the "reply" function in your email does *not* send your answer
to the person asking the question.
Please copy their email address from their question.
****************************************************************************************
realname - Purusharth
Email - rajyagur-at-email.arizona.edu
EDUCATION - Graduate College
LOCATION - TUCSON
SUBJECT_OF_QUESTION - Yeast live cell imaging
QUESTION - I am interested in imaging live yeast cells expressing
fluorescent tagged proteins. My impression is that deltavision RT scope
(Applied precision) is the best option for doing that. Is that right?

1. Why does confocal microscope does not give equally good result as
deltavision in above case?

2. Would any other wide-filed fluorescent microscope suffice for above
use (would be preferred for it cheaper cost than deltavision)?

thanks for your help.

best
raj

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Title-Subject: [Filtered] Job - Research Associate I -at- University of Delaware

Message: Research Associate I Advanced Microscopy and Microanalysis

DEADLINE: March 15, 2013

QUALIFICATIONS:Bachelor's degree in physical, chemical, biological, or a related field and one year
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MAJOR RESPONSIBILITIES:

Trains and supervises students and post-docs in utilizing SEM/FIB, TEM, LM, SPM and auxiliary
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Tom:

Where as I have users who do perfusion fixation I rarely do, but i had
to speak to your buffer question. I almost always use Na cacodylate for
immersion fixation, but I would never use Na cacodylate for perfusion
fixation. Na cacodylate is arsenic based and is toxic the tissue will respond
to the arsenic toxicity, potentially adding artifacts. When I have perfusion
fixed the goal is to keep the tissue as normally happy as possible, simply
cross-linking the protein and immobilizing and stabilizing the cell systems in
situ. And again avoiding the autolytic \ injury response artifacts that go with
the time required for hacking apart the tissue to get at the desired regions.
Also keep in mind the effects of any anesthetics may have on the tissue.

(And yes the aldehydes are toxins to but we can't really avoid them
unless we go cryo. . . . in which case we're back to tissue degradation due to
excision)

On 25 Feb 2013 at 16:54, tbargar-at-unmc.edu wrote:

}
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} Dear Listers,
}
} In fixing brain tissue, is one type of buffer better than any other.
} Here are the conditions, perfusion fixation, 2.5% glutaraldehyde + 2%
} paraformaldehyde, pH 7.2 to 7.4. Buffers in question are .1m Sodium
} Phosphate buffer vs. .1M Sodium Cacodylate buffer. Is one better than
} the other? Also could there be some other buffer that is better? I
} am not involved in a project involving brain tissue, but I have been
} asked my opinion. In my experience I have seen neural tissues that
} have been fixed in fixatives that have used both types of buffers and
} the fixation in any of those tissues was fine. However to provide the
} individual with as much advice as possible, I would like to pass on
} the advice and opinions of others, especially anyone out there who has
} a lot of experience with neural tissue. As always thanks in advance.
}
} Tom Bargar
} UNMC
} Electron Microscopy Core Facility
} 986395 Nebraska Medical Center
} Omaha, NE 68198-6395
} tbargar-at-unmc.edu
} 402-559-7347
}
}
}
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Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
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Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-muohio.edu
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Hi All,

I have two requests for information and decided to combine in one E-mail
to spare your inboxes.

1. I would like to hear from those of you who can recommend a vendor for a
low background holder for an FEI Titan. This is needed after install of
an EDS system. Any personal preferences, recommendations or concerns about
holders or vendors would be appreciated
but probably best answered off line.
2. Also I have a colleague looking for recommendations for a service
engineer in the New England area to service an FEI Quanta SEM. Please
contact me if you can recommend someone or if you are the service provider
and I will pass on the information.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Email: hguthrey-at-mines.edu
Name: Harvey Guthrey

Organization: Colorado School of Mines

Title-Subject: [Filtered] Critical vacuum level to avoid hydrocarbon contamination while coolint to 20K

Message: I have an SEM equipped with a LHe cryostat that is used for CL spectroscopy at
approximately 20K. It takes about 5 hours to reach 20K from 300K. It also takes around 4 hours to
achieve ultimate vacuum in the chamber(9.6 E-5 Pa is the lower limit of the gauge). If I turn on
the cryostat soon after loading the sample there is noticeable HC contamination that significantly
alters the CL from the sample. Does anyone have an idea of what range of chamber vacuum would be
appropriate for us to begin cooling the sample without having it behave like a cold trap? This
would allow us to begin analysis in less than 9 hours as is our current situation. Thanks

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Dear Harvey,

Based on the ungodly pumping time of 4 hours and level of hydrocarbon
contamination that you are describing, chances are that your instrument
is severely contaminated, with most likely sources of such contamination
being non-vacuum-compatible oil used for lubricating state and/or
poor-quality roughing pump oil back-streamed into main chamber.

The way to resolve this would be to take apart vacuum system, thoroughly
clean it with toluene followed by acetone+alcohol, discard all the
O-rings with new ones (Duniway Stockroom is supplying pre-baked O-rings,
no interest - just a happy customer), religiously clean stage and
lubricate it with y25-9 or better oil, install alumina trap on the
roughing pump or use dry mechanical pump, and install Evactron or
similar device to pre-clean chamber for each sample since no matter what
you do to clean severely contaminated system oils will continue
outgassing from crevices in the stage for quite a few years. If there is
a diffusion pump, make sure that it uses oil compatible with UHV range,
or better yet replace it with turbo.

If you have spare ports on the chamber, you can use them to put in a
couple of cold fingers to help trapping some of the oil molecules as a
band-aid.

Providing that you can't source hardware expertize to clean and dry the
vacuum system and/or money to hire such expertize and pay for
pump/trap/Evactron/cold fingers, then the only (known to me) realistic
alternative is to wait patiently for system to reach ultimate vacuum and
then start cooling, which you are doing already...

Good luck :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
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Web: www.freudlabs.com

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} Title-Subject: [Filtered] Critical vacuum level to avoid hydrocarbon contamination while coolint to 20K
}
} Message: I have an SEM equipped with a LHe cryostat that is used for CL spectroscopy at
} approximately 20K. It takes about 5 hours to reach 20K from 300K. It also takes around 4 hours to
} achieve ultimate vacuum in the chamber(9.6 E-5 Pa is the lower limit of the gauge). If I turn on
} the cryostat soon after loading the sample there is noticeable HC contamination that significantly
} alters the CL from the sample. Does anyone have an idea of what range of chamber vacuum would be
} appropriate for us to begin cooling the sample without having it behave like a cold trap? This
} would allow us to begin analysis in less than 9 hours as is our current situation. Thanks
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Email: liliana-at-unimelb.edu.au
Name: Liliana

Organization: School of Veterinary Science University of Melbourne

Title-Subject: [Filtered] Philips 505 SEM

Message: I am hoping someone on this forum has access to the operational and technical manuals for a
Philips 505 SEM. Our engineer is happy to service the microscope, but he needs the technical
specifications in order to troubleshoot effectively. If anybody has this information available, it
would be extremely helpful. Thank you very much for your assistance.

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Name: James Fitzpatrick

Organization: Salk Institute for Biological Studies

Title-Subject: [Filtered] Re: Salk Institute Imaging Symposium - May 31st 2013

Message: Dear Microscopy Colleagues,

On May 31, 2013, the WAITT ADVANCED BIOPHOTONICS CENTER at the Salk Institute will be hold its
SECOND ANNUAL IMAGING SYMPOSIUM:

"SINGLE MOLECULES TO CELLULAR SYSTEMS: JUGGLING COMPLEXITY"

Please see the symposium website for the scientific program: http://www.salk.edu/wabc2013

POSTER ABSTRACT SUBMISSION DEADLINE : APRIL 15th 2013

The organizing committee will be selecting 30 posters for exhibition in the lunchtime poster session
from the submitted abstracts. Please note that you must also register for the symposium if you want
to submit an abstract. Submitting an abstract alone does not guarantee registration.

http://SalkInstitute.kintera.org/WABC2013abstract

REGISTRATION DEADLINE : May 13th 2013

Registration is free for all Salk scientists and includes lunch. YOU MUST REGISTER EVEN IF YOU
SUBMITTED AN ABSTRACT!!

http://SalkInstitute.kintera.org/WABC2013

We look forward to seeing you all there.

WABC 2013 Symposium Organizing Committee
Martin Hetzer (Faculty Director, WABC)
Axel Nimmerjahn (Assistant Professor, WABC)
James Fitzpatrick (Core Director, WABC)

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Ropa-a-dope only got me so far and then management scored a clear hit with the question "How do other TEM labs calibrate their microscope?"

I wanted to fire up the Lear jet and start my survey somewhere nice, but instead I'm turning to the list server for help.

Hopefully this will not be too painful.

I would like to know is:

What do you calibrate your TEM with for linear measurements?
Do you use one standard over your entire range of magnification?
Is your lab certified by some organization for TEM use and if so by who?
And finally, if it's not too nosy, how often do you calibrate your scope?

When I get all the results, I'll send a short summary out and have the details available to anyone who wants them.

Full disclosure: ARDL is a commercial lab and we want to provide the best possible results to our customers. We calibrate our TEM monthly and measure a population of 203nm NIST certified microspheres as verification.

Thanks!!!!!

Frank Karl

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Hi Harvey,
As Valery alluded to, the key factor is the partial pressure of
hydrocarbons. These cause all the problems even when they contribute
negligibly to the overall pressure. Cleaning the system and regularly using
a de-contaminator (while costly) is the best way to eliminate the problem.
Chamber plasma cleaners will allow even cryo-imaging, and even in non-UHV
conditions. Good luck!

Best Regards,
Larry Scipioni
--
Larry Scipioni, PhD
Vice President of R&D - Platform Development
les-at-zsgenetics.com
781-552-1989 (mobile)
larry.scipioni (Skype)
ZS Genetics
www.zsgenetics.com

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Email: hguthrey-at-mines.edu
Name: Harvey Guthrey

Organization: Colorado School of Mines

Title-Subject: [Filtered] Critical vacuum level to avoid hydrocarbon
contamination while coolint to 20K

Message: I have an SEM equipped with a LHe cryostat that is used for CL
spectroscopy at approximately 20K. It takes about 5 hours to reach 20K from
300K. It also takes around 4 hours to achieve ultimate vacuum in the
chamber(9.6 E-5 Pa is the lower limit of the gauge). If I turn on the
cryostat soon after loading the sample there is noticeable HC contamination
that significantly alters the CL from the sample. Does anyone have an idea
of what range of chamber vacuum would be appropriate for us to begin cooling
the sample without having it behave like a cold trap? This would allow us
to begin analysis in less than 9 hours as is our current situation. Thanks

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Dear All,
I am looking for user manuals and if available, service manuals for the Reichert OM U3 ultramicrotome.
If anybody has them in PDF and could send them over, this would be perfect.

Best wishes,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
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The NUANCE Center from Northwestern University has put out a new episode of the web series What's It Made Of? This is going to be a monthly series demonstrating the SEM and the kinds of things you can do with it. It's fun, surprisingly sort-of educational and appropriate for all age groups. (Except maybe 0-2 years of age.) If you like it, we hope you can spread it around!

http://www.youtube.com/watch?v=aJPOTRe0Hxg

ERiC Jay Miller
Microscopy & Imaging Specialist
Electron Probe Instrumentation Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1152 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 467-0789
fax: (847) 467-657

http://www.nuance.northwestern.edu

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On Mar 8, 2013, at 5:21 AM, frank_karl-at-ardl.com wrote:

} I would like to know is:
}
} What do you calibrate your TEM with for linear measurements?
} Do you use one standard over your entire range of magnification?
} Is your lab certified by some organization for TEM use and if so by
} who?
} And finally, if it's not too nosy, how often do you calibrate your
} scope?
}
Dear Frank,
I used the Mag*I*Cal on the HVEM, but the FEI camera calibration
requires a cross-grating replica. Either is suitable for linear
measurements, and, if you can find gold lattice lines on the cross-
grating, it is OK for pretty much all mags (except the low end of low
mag range). The Mag*I*Cal can be used for all mags and camera
lengths, Neither of the labs for which I calibrated the mag was
certified, and we calibrated the HVEM once a year plus when a user
wanted absolute dimensions. The FEI scopes were calibrated more
often, since some of the software we used required calibration during
set-up. In that case, we just ran through all the required
calibrations, which took about a half hour.
Yours,
Bill

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Name: Mark Aindow

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Title-Subject: [Filtered] Postdoctoral Position

Message: UNIVERSITY OF CONNECTICUT
INSTITUTE OF MATERIALS SCIENCE

Postdoctoral Position in Materials for Circuit Breaker Contacts

A post-doctoral position is available immediately within the Institute
of Materials Science at the University of Connecticut to work on
metallic alloys and metal-matrix composites for electrical circuit
breaker contacts. This project is an industrial collaboration sponsored
by GE Energy Industrial Solutions of Plainville, CT. Candidates should
have a strong background in physical metallurgy and experience in
microstructural characterization (particularly transmission electron
microscopy). Prior experience in electrical property measurements and
computational modeling (first principles and/or thermodynamic) would
also be beneficial. This position is a one-year appointment, renewable
for a second year.

To apply please send: a resume, a list of publications, and contact
details for 3 references to Dr. Mark Aindow (m.aindow-at-uconn.edu).
Screening of applications will begin immediately, and will continue
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Title-Subject: [Filtered] Suggestions for donation of older microscopes?

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Quick reminder that the registration deadline for the 3rd UB Practical
Course on Cryo-Electron Microscopy of Vitreous Sections is on
17.3.2013

Regards
Benoit Zuber

---------- Forwarded message ----------
X-from: Beno�t Zuber {benoit.zuber.work-at-gmail.com}

Dear all,

We are pleased to announce

the 3rd UB Practical Course on Cryo-Electron Microscopy of Vitreous
Sections (CEMOVIS, a.k.a cryoEM of frozen-hydrated sections)

2-4 July 2013 / 9-11 July 2013

Organiser : Beno�t Zuber
Instructors: Beno�t Zuber, Daniel Studer, Ioan Iacovache

The objective of the course is to teach participants the practical
skills necessary to successfully apply CEMOVIS (a.k.a. Frozen-hydrated
sections) in their laboratories. Our latest tricks making CEMOVIS
quite easy will be demonstrated. Essential background theory of
CEMOVIS will be given and most of the time will be spent practicing
high-pressure freezing, cryo-sectioning, and low-dose TEM imaging of
vitreous sections. 1 high-pressure freezing machine, 3
state-of-the-art cryo-ultramicrotomes and 1 cryo-electron microscope
will be dedicated to the course. The 3-day course is given twice for
two different groups of maximum 4 participants each so that every
participant can be actively practicing during the whole course.

All the participants of the previous previous courses have been
successful in producing, collecting and imaging vitreous sections.

The course is intended for scientists whose research projects will
benefit from the use of CEMOVIS. Experience in either cryo-electron
microscopy or ultramicrotomy of resin-embedded specimens is a
prerequisite.

Information and registration:
http://www.ana.unibe.ch/events/cemovis/index.html

Registration deadline: 17.03.2013

Yours faithfully
Beno�t Zuber
Note: do not reply to this e-mail address but to the one in my signature below.
--
Prof. Beno�t Zuber
Institute of Anatomy
University of Bern
Baltzerstrasse 2
3000 Bern 9
Switzerland
benoit.zuber-at-ana.unibe.ch
+41 31 631 84 40

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Name: Tina Williams

Organization: USDA

Title-Subject: [Filtered] Suggestions for Donations of Older
Microscopes-Thank you - Microscopes are all placed

Message: Hello,

Thank you to all who replied with suggestions on where/how to donate
older microscopes. The researcher has found places for all of the
microscopes.

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With the move to a new lab and the installation of a new instrument I have been performance testing all the microscopes. I also have an upcoming project which will need some careful measurement of topography so need to know the tilt angle accuracy and repeatability.

I thought no problem since I have circles on a Geller MRS-4 standard. A circle at tilt becomes an ellipse and one can use the ratio of max length/min width to calculate the perspective and camera angle, common in scientific photography.

My math is sound because I can use the same standard on the light microscope with a known wedge angle and I measure correctly and am within 1 degree both with automated software measurements and by hand with Photoshop.

But on the SEM, very carefully leveling the standard, correcting astigmatism, using a moderately fast scan rate to minimize drift errors, and taking into account the XY calibration ratios of the scan I am still seeing 5 or more degrees of variance.

Is this the best I can expect or is there a better way to do this?

Thanks,

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012�� MRC104
Washington DC 20013-7012
202-633-0891

The move is complete. The lab is now located in W150- First floor west wing- northwest corner.

==============================Original Headers==============================
11, 26 -- From WHITTAKS-at-si.edu Wed Mar 13 09:31:32 2013
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If you have a recent model Zeiss EVO, and specifically the LS/MA15 large stage machine I would appreciate corresponding off line with you.

Thanks

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012�� MRC104
Washington DC 20013-7012
202-633-0891

The move is complete. The lab is now located in W150- First floor west wing- northwest corner.

==============================Original Headers==============================
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Hi Scott,
I would think that 5˚ would be really obvious in looking at the stage but if you're getting the angle from x vs y, that's going to be really touchy around 0˚, so I'd suspect that your x and y drives aren't stable (or consistent) or you are getting stage drift, thermal or otherwise in one axis or the other.

I guess I'm saying that the stage tilt should be fairly accurate and reproducible at the 1˚ level, so I think the problem is elsewhere. See if the accuracy and/or reproducibility is better around 45˚, where there is less sensitivity to x vs y because the changes are greater.

One other thing to check would be the stage hysteresis for tilt. Come at 0˚ first from one direction, then the other and see how your measurements compare. Hopefully there isn't 5˚ slop, but you never know.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

-----Original Message-----
X-from: WHITTAKS-at-si.edu [mailto:WHITTAKS-at-si.edu]
Sent: Wednesday, March 13, 2013 10:35 AM
To: kenconverse-at-qualityimages.biz

With the move to a new lab and the installation of a new instrument I have been performance testing all the microscopes. I also have an upcoming project which will need some careful measurement of topography so need to know the tilt angle accuracy and repeatability.

I thought no problem since I have circles on a Geller MRS-4 standard. A circle at tilt becomes an ellipse and one can use the ratio of max length/min width to calculate the perspective and camera angle, common in scientific photography.

My math is sound because I can use the same standard on the light microscope with a known wedge angle and I measure correctly and am within 1 degree both with automated software measurements and by hand with Photoshop.

But on the SEM, very carefully leveling the standard, correcting astigmatism, using a moderately fast scan rate to minimize drift errors, and taking into account the XY calibration ratios of the scan I am still seeing 5 or more degrees of variance.

Is this the best I can expect or is there a better way to do this?

Thanks,

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012 MRC104
Washington DC 20013-7012
202-633-0891

The move is complete. The lab is now located in W150- First floor west wing- northwest corner.

==============================Original Headers==============================
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Hi Scott,
What is the angle of your SEM detector that you are using to measure the circles? I believe that SEM detectors are at some angle to the specimen, probably even true for "in-lens detectors". This could account for the angle observed.

Roseann Csencsits, PhD
Scientist in Charge, Downing TEM Tools
Lawrence Berkeley Lab 01-365
1 Cyclotron Road
Berkeley, CA 94720
510-486-4548

On Mar 13, 2013, at 7:43 AM, WHITTAKS-at-si.edu wrote:

}
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} With the move to a new lab and the installation of a new instrument I have been performance testing all the microscopes. I also have an upcoming project which will need some careful measurement of topography so need to know the tilt angle accuracy and repeatability.
}
} I thought no problem since I have circles on a Geller MRS-4 standard. A circle at tilt becomes an ellipse and one can use the ratio of max length/min width to calculate the perspective and camera angle, common in scientific photography.
}
} My math is sound because I can use the same standard on the light microscope with a known wedge angle and I measure correctly and am within 1 degree both with automated software measurements and by hand with Photoshop.
}
} But on the SEM, very carefully leveling the standard, correcting astigmatism, using a moderately fast scan rate to minimize drift errors, and taking into account the XY calibration ratios of the scan I am still seeing 5 or more degrees of variance.
}
} Is this the best I can expect or is there a better way to do this?
}
} Thanks,
}
} Scott Whittaker
} Manager SEM Lab
} Smithsonian Institution
} National Museum of Natural History
} PO Box 37012 MRC104
} Washington DC 20013-7012
} 202-633-0891
}
} The move is complete. The lab is now located in W150- First floor west wing- northwest corner.
}
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Ken,

Thanks for the input. Indeed I have leveled the instrument, and drawn a line on the monitor of the chamber camera and can visually detect {1 degree tilt. Resetting the next run I used a bubble level on the instrument and matched another on the surface of the loaded sample to make sure it was as close to machine square as possible. In doing so I have now squared the machine.

I am getting similar results from the other instrument and it seems even 1 pixel deviation in my measuring is significant, but since I can do this at the light level with just a penny on a known tilt I am confident in my measurement workflow on a traceable standard.

For the next run I intend to make a set of self referencing Teflon feet and run the stage up onto the lens to ensure the column/ stage is absolutely perpendicular and see if it makes a difference in my readings. This should square it, and help the mechanical drift I hope. I am somewhat mistrustful of anything until I can get 0 degrees to measure 0 degrees and I suppose this will be the reported error I use.

Stage drift is definitely an issue, but based on earlier data I obtained measuring the drift rate during the performance testing the fast scan speed was chosen to be 1 pixel or less during capture at 968 lines. Image was noisy thus my decision to check the automated analysis with hand measurements via Photoshop.

I hadn't considered thermal drift and just got all excited, but it seems that thermal drift would also be a part of the stage drift testing results and thus should have been part of the fast scan rate selection.

I am out of ideas here and thank you for the valuable input. Who knew you could blow an entire day just checking the tilt angle reported on the stages.

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012 MRC104
Washington DC 20013-7012
202-633-0891

The move is complete. The lab is now located in W150- First floor west wing- northwest corner.

-----Original Message-----
X-from: Ken Converse [mailto:kenconverse-at-qualityimages.biz]
Sent: Wednesday, March 13, 2013 11:10 AM
To: Whittaker, Scott; MSA Listserver

With the move to a new lab and the installation of a new instrument I have been performance testing all the microscopes. I also have an upcoming project which will need some careful measurement of topography so need to know the tilt angle accuracy and repeatability.

I thought no problem since I have circles on a Geller MRS-4 standard. A circle at tilt becomes an ellipse and one can use the ratio of max length/min width to calculate the perspective and camera angle, common in scientific photography.

My math is sound because I can use the same standard on the light microscope with a known wedge angle and I measure correctly and am within 1 degree both with automated software measurements and by hand with Photoshop.

But on the SEM, very carefully leveling the standard, correcting astigmatism, using a moderately fast scan rate to minimize drift errors, and taking into account the XY calibration ratios of the scan I am still seeing 5 or more degrees of variance.

Is this the best I can expect or is there a better way to do this?

Thanks,

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012 MRC104
Washington DC 20013-7012
202-633-0891

The move is complete. The lab is now located in W150- First floor west wing- northwest corner.

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X-from: michael.cammer-at-med.nyu.edu ()

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Email: michael.cammer-at-med.nyu.edu
Name: Michael Cammer

Organization: NYU

Title-Subject: [Filtered] Nunc Labtek chamber question

Message: Hi.

For many years we used Nunc Lab-Tek chambered coverglasses. They were great.

However, they recently began shipping a different design with the same
product number. The chambers no longer fit in our microscope stages
inserts. Also, the volume changed.
see http://www.flickr.com/photos/mcammer/sets/72157632984171809/

While we can adjust to the volume change, the plastic tab at the end of
the chamber is a big problem.

I thought of having people cut the tab off before doing experiments, but
this would produce a lot of plastic dust (and we'd have to buy a
suitable saw or dremel).

Does anybody know of an alternative manufacturer or if the old style
chambers are still available from somewhere?

Thanks!

Michael

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I guess I should clear up that I haven't gotten around to quantifying my Zeiss instrument yet. My stage posting is unrelated to the Zeiss users request. As far as I know I do not have stage issues on my Zeiss though I have only measured drift rate to date.

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012�� MRC104
Washington DC 20013-7012
202-633-0891

The move is complete. The lab is now located in W150- First floor west wing- northwest corner.

X-from: Keith Collins [mailto:Keith.Collins-at-NETL.DOE.GOV]
Sent: Wednesday, March 13, 2013 11:07 AM
To: Whittaker, Scott

If you have a recent model Zeiss EVO, and specifically the LS/MA15 large stage machine I would appreciate corresponding off line with you.

Thanks

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012�� MRC104
Washington DC 20013-7012
202-633-0891

The move is complete. The lab is now located in W150- First floor west wing- northwest corner.

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Hello Scott,

May you tell us more about your instrument ? Motorized stage or not,
model of the SEM ? Do you use stereoscopy software ?
Nicolas Stephant
University of Nantes
France

WHITTAKS-at-si.edu a �crit :
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} Ken,
}
} Thanks for the input. Indeed I have leveled the instrument, and drawn a line on the monitor of the chamber camera and can visually detect {1 degree tilt. Resetting the next run I used a bubble level on the instrument and matched another on the surface of the loaded sample to make sure it was as close to machine square as possible. In doing so I have now squared the machine.
}
} I am getting similar results from the other instrument and it seems even 1 pixel deviation in my measuring is significant, but since I can do this at the light level with just a penny on a known tilt I am confident in my measurement workflow on a traceable standard.
}
} For the next run I intend to make a set of self referencing Teflon feet and run the stage up onto the lens to ensure the column/ stage is absolutely perpendicular and see if it makes a difference in my readings. This should square it, and help the mechanical drift I hope. I am somewhat mistrustful of anything until I can get 0 degrees to measure 0 degrees and I suppose this will be the reported error I use.
}
} Stage drift is definitely an issue, but based on earlier data I obtained measuring the drift rate during the performance testing the fast scan speed was chosen to be 1 pixel or less during capture at 968 lines. Image was noisy thus my decision to check the automated analysis with hand measurements via Photoshop.
}
} I hadn't considered thermal drift and just got all excited, but it seems that thermal drift would also be a part of the stage drift testing results and thus should have been part of the fast scan rate selection.
}
} I am out of ideas here and thank you for the valuable input. Who knew you could blow an entire day just checking the tilt angle reported on the stages.
}
}
} Scott Whittaker
} Manager SEM Lab
} Smithsonian Institution
} National Museum of Natural History
} PO Box 37012� � MRC104
} Washington DC 20013-7012
} 202-633-0891
}
} The move is complete. The lab is now located in W150- First floor west wing- northwest corner.
}
}
} -----Original Message-----
} X-from: Ken Converse [mailto:kenconverse-at-qualityimages.biz]
} Sent: Wednesday, March 13, 2013 11:10 AM
} To: Whittaker, Scott; MSA Listserver
} Subject: RE: [Microscopy] SEM tilt calibration
}
} Hi Scott,
} I would think that 5˚ would be really obvious in looking at the stage but if you're getting the angle from x vs y, that's going to be really touchy around 0˚, so I'd suspect that your x and y drives aren't stable (or consistent) or you are getting stage drift, thermal or otherwise in one axis or the other.
}
} I guess I'm saying that the stage tilt should be fairly accurate and reproducible at the 1˚ level, so I think the problem is elsewhere. See if the accuracy and/or reproducibility is better around 45˚, where there is less sensitivity to x vs y because the changes are greater.
}
} One other thing to check would be the stage hysteresis for tilt. Come at 0˚ first from one direction, then the other and see how your measurements compare. Hopefully there isn't 5˚ slop, but you never know.
}
} Ken Converse
} owner
}
} QUALITY IMAGES
} Servicing Scanning Electron Microscopes
} Since 1981
} 474 So. Bridgton Rd.
} Bridgton, ME 04009
} 207-647-4348
} kenconverse-at-qualityimages.biz
} qualityimages.biz
}
}
} -----Original Message-----
} X-from: WHITTAKS-at-si.edu [mailto:WHITTAKS-at-si.edu]
} Sent: Wednesday, March 13, 2013 10:35 AM
} To: kenconverse-at-qualityimages.biz
} Subject: [Microscopy] SEM tilt calibration
}
}
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} With the move to a new lab and the installation of a new instrument I have been performance testing all the microscopes. I also have an upcoming project which will need some careful measurement of topography so need to know the tilt angle accuracy and repeatability.
}
} I thought no problem since I have circles on a Geller MRS-4 standard. A circle at tilt becomes an ellipse and one can use the ratio of max length/min width to calculate the perspective and camera angle, common in scientific photography.
}
} My math is sound because I can use the same standard on the light microscope with a known wedge angle and I measure correctly and am within 1 degree both with automated software measurements and by hand with Photoshop.
}
} But on the SEM, very carefully leveling the standard, correcting astigmatism, using a moderately fast scan rate to minimize drift errors, and taking into account the XY calibration ratios of the scan I am still seeing 5 or more degrees of variance.
}
} Is this the best I can expect or is there a better way to do this?
}
} Thanks,
}
} Scott Whittaker
} Manager SEM Lab
} Smithsonian Institution
} National Museum of Natural History
} PO Box 37012 MRC104
} Washington DC 20013-7012
} 202-633-0891
}
} The move is complete. The lab is now located in W150- First floor west wing- northwest corner.
}
}
}
}
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--
Nicolas STEPHANT

Universit� de Nantes
Institut Jean Rouxel
Service de microscopie �lectronique � balayage et microanalyse
2 rue de la Houssini�re
BP 92208
44322 Nantes c�dex 3

T�l : 02 40 37 64 26
M�l : nicolas.stephant-at-univ-nantes.fr
Web : http://www.univ-nantes.fr/smebm

"Le monde n'existe que pour autant que nous sommes capables d'en
produire une image"
C.G Jung

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Hi Scott,
Just out of curiosity, how much change in tilt does a one-pixel change give you? This would be a 0.1% error in linear measurement at 1000 lines. This level of accuracy is generally considered beyond the scope of SEMs unless it is a CD (critical dimension) instrument, although repeatability (precision) should be in that range, given identical working conditions such as working distance and a lens hysteresis procedure to normalize the lenses (and therefore the WD).

When you're doing this with a light microscope, what are the critical parameters of the image you are working with? Is it simply from an NTSC camera, a hi resolution camera, or scanned from film? What kind of repeatability do you get?

Try taking consecutive images of your standard on the SEM, then make all the changes and adjustments that you normally would except don't actually change the tilt. Take another image and see how it compares to the first two that were taken right after one another. Changes in lens hysteresis cause apparent changes in WD which is a direct factor in calculating the magnification.

If you focus, then go to a very long WD, refocus, take an image, go to a very short WD, refocus and take an image, I think you will find that the images may be identical, but the indicated mag will be quite different, or, if you set the mag the same, the images will be different sizes. This may be at least part of the problem with these very fine measurements. This is why a standard procedure for normalizing hysteresis is necessary. Some instruments have this built in (sometimes as an automated routine, sometimes as a lens reversal button), others don't.

On older JEOLs and Amrays, there was a lens current reversal button. I found that pressing it for less than a second gave variable results, while pressing for a second or more gave much more consistent results. Newer JEOLs have a computer controlled routine that is very consistent. On ETECs, the KV select button was undepressed, which turned off the lenses. I found that less that 15 seconds gave highly variable results. More than 15 seconds was more stable, but using the same time each time was important, so I always used 2 sweeps at the SYNC speed which was 16 seconds total and got consistent results for calibrating.

It may be that you will want to use a different technique for measuring the topography in your upcoming project, if a high degree of accuracy is needed. On the other hand, if your image size (mag) is consistent, I would think that the stage tilt readout should give you enough accuracy for your stereo pairs and subsequent processing.

Lots of variables here :-)

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

-----Original Message-----
X-from: Whittaker, Scott [mailto:WHITTAKS-at-si.edu]
Sent: Wednesday, March 13, 2013 1:19 PM
To: 'Ken Converse'
Cc: 'microscopy-at-microscopy.com'

With the move to a new lab and the installation of a new instrument I have been performance testing all the microscopes. I also have an upcoming project which will need some careful measurement of topography so need to know the tilt angle accuracy and repeatability.

I thought no problem since I have circles on a Geller MRS-4 standard. A circle at tilt becomes an ellipse and one can use the ratio of max length/min width to calculate the perspective and camera angle, common in scientific photography.

My math is sound because I can use the same standard on the light microscope with a known wedge angle and I measure correctly and am within 1 degree both with automated software measurements and by hand with Photoshop.

But on the SEM, very carefully leveling the standard, correcting astigmatism, using a moderately fast scan rate to minimize drift errors, and taking into account the XY calibration ratios of the scan I am still seeing 5 or more degrees of variance.

Is this the best I can expect or is there a better way to do this?

Thanks,

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012 MRC104
Washington DC 20013-7012
202-633-0891

The move is complete. The lab is now located in W150- First floor west wing- northwest corner.

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Hi All,

Have you found significant inaccuracies with X-ray mapping when using a
very high end FE-TEM (such as FEI Titan) and tilting the sample to the
angle required for maximum X-ray counts? To my knowledge, the position of
many EDS collimators requires a tilt
of 20-30 degrees in order to obtain a sufficient count to offset
background. The tilting does mean that the beam is traveling through a
thicker and angled area of the sample. My concern would be in situations
like mapping locations of molecules coating nano
particles or other instances where very accurate location of elements
relative to other nano features is desired.

Does the tilting to this amount cause sufficient offset and location error
to be a concern with very high resolution instruments?
How do you handle such situations if you do think it makes a significant
difference?
Are there some preferred detector/microscope criteria or combinations that
reduce this problem and should be considered when purchasing an EDS system
for this level of instrument? (vendor comments appreciated but use
discretion as to whether they should
be off line).

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Hello Scott,

would the tilt sensors like those in the MIAWiki page:
http://confocal-manawatu.pbworks.com/w/page/64499329/Precision%20Tilt%20Sensors
make your work faster next time? The angular accuracy in XY plane seems being well below 1 degree.

Cheers,
Dmitry

On 14/03/2013 10:40 p.m., Nicolas.Stephant-at-univ-nantes.fr wrote:
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} Hello Scott,
}
} May you tell us more about your instrument ? Motorized stage or not,
} model of the SEM ? Do you use stereoscopy software ?
} Nicolas Stephant
} University of Nantes
} France
}
} WHITTAKS-at-si.edu a �crit :
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} }
} } Ken,
} }
} } Thanks for the input. Indeed I have leveled the instrument, and drawn a line on the monitor of the chamber camera and can visually detect {1 degree tilt. Resetting the next run I used a bubble level on the instrument and matched another on the surface of the loaded sample to make sure it was as close to machine square as possible. In doing so I have now squared the machine.
} }
} } I am getting similar results from the other instrument and it seems even 1 pixel deviation in my measuring is significant, but since I can do this at the light level with just a penny on a known tilt I am confident in my measurement workflow on a traceable standard.
} }
} } For the next run I intend to make a set of self referencing Teflon feet and run the stage up onto the lens to ensure the column/ stage is absolutely perpendicular and see if it makes a difference in my readings. This should square it, and help the mechanical drift I hope. I am somewhat mistrustful of anything until I can get 0 degrees to measure 0 degrees and I suppose this will be the reported error I use.
} }
} } Stage drift is definitely an issue, but based on earlier data I obtained measuring the drift rate during the performance testing the fast scan speed was chosen to be 1 pixel or less during capture at 968 lines. Image was noisy thus my decision to check the automated analysis with hand measurements via Photoshop.
} }
} } I hadn't considered thermal drift and just got all excited, but it seems that thermal drift would also be a part of the stage drift testing results and thus should have been part of the fast scan rate selection.
} }
} } I am out of ideas here and thank you for the valuable input. Who knew you could blow an entire day just checking the tilt angle reported on the stages.
} }
} }
} } Scott Whittaker
} } Manager SEM Lab
} } Smithsonian Institution
} } National Museum of Natural History
} } PO Box 37012� � MRC104
} } Washington DC 20013-7012
} } 202-633-0891
} }
} } The move is complete. The lab is now located in W150- First floor west wing- northwest corner.
} }
} }
} } -----Original Message-----
} } X-from: Ken Converse [mailto:kenconverse-at-qualityimages.biz]
} } Sent: Wednesday, March 13, 2013 11:10 AM
} } To: Whittaker, Scott; MSA Listserver
} } Subject: RE: [Microscopy] SEM tilt calibration
} }
} } Hi Scott,
} } I would think that 5˚ would be really obvious in looking at the stage but if you're getting the angle from x vs y, that's going to be really touchy around 0˚, so I'd suspect that your x and y drives aren't stable (or consistent) or you are getting stage drift, thermal or otherwise in one axis or the other.
} }
} } I guess I'm saying that the stage tilt should be fairly accurate and reproducible at the 1˚ level, so I think the problem is elsewhere. See if the accuracy and/or reproducibility is better around 45˚, where there is less sensitivity to x vs y because the changes are greater.
} }
} } One other thing to check would be the stage hysteresis for tilt. Come at 0˚ first from one direction, then the other and see how your measurements compare. Hopefully there isn't 5˚ slop, but you never know.
} }
} } Ken Converse
} } owner
} }
} } QUALITY IMAGES
} } Servicing Scanning Electron Microscopes
} } Since 1981
} } 474 So. Bridgton Rd.
} } Bridgton, ME 04009
} } 207-647-4348
} } kenconverse-at-qualityimages.biz
} } qualityimages.biz
} }
} }
} } -----Original Message-----
} } X-from: WHITTAKS-at-si.edu [mailto:WHITTAKS-at-si.edu]
} } Sent: Wednesday, March 13, 2013 10:35 AM
} } To: kenconverse-at-qualityimages.biz
} } Subject: [Microscopy] SEM tilt calibration
} }
} }
} }
} }
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} } With the move to a new lab and the installation of a new instrument I have been performance testing all the microscopes. I also have an upcoming project which will need some careful measurement of topography so need to know the tilt angle accuracy and repeatability.
} }
} } I thought no problem since I have circles on a Geller MRS-4 standard. A circle at tilt becomes an ellipse and one can use the ratio of max length/min width to calculate the perspective and camera angle, common in scientific photography.
} }
} } My math is sound because I can use the same standard on the light microscope with a known wedge angle and I measure correctly and am within 1 degree both with automated software measurements and by hand with Photoshop.
} }
} } But on the SEM, very carefully leveling the standard, correcting astigmatism, using a moderately fast scan rate to minimize drift errors, and taking into account the XY calibration ratios of the scan I am still seeing 5 or more degrees of variance.
} }
} } Is this the best I can expect or is there a better way to do this?
} }
} } Thanks,
} }
} } Scott Whittaker
} } Manager SEM Lab
} } Smithsonian Institution
} } National Museum of Natural History
} } PO Box 37012 MRC104
} } Washington DC 20013-7012
} } 202-633-0891
} }
} } The move is complete. The lab is now located in W150- First floor west wing- northwest corner.
} }
} }
} }
} }
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Hi Dan,

Hi Dan,

I use a Philips CM200 FEG. I have found the sample tilt is indeed very important in TEM/EDS quants. Even a tilt of only 20 degrees is a pretty small takeoff angle, and TEM samples are often not flat. Also, as you say, the spatial resolution of the TEM can make offset issues troublesome when measuring small grains. Also, make sure your k-factors are measured at the same tilt and beam conditions.

I do the following, in order of importance, to get get good quants:

After centering your sample, and focusing, etc. Wait about 2-3 minutes for the stage to really stop drifting. Then make your measurement, or use a drift correction with a short turnaround (e.g. 20 seconds if you want to take spectra right after moving the goniometer).

Some samples burn easily. Take a sequence of spectra each about 30 seconds long (or one minute, etc, depending on your beam intensity and count rate). Compare them. If they are different your sample is changing! You can solve that by using a larger raster area or noticing that it is just one element that varies, and then use other methods to quantify it (I've used STXM, stoichometry, backtracking a sequence of spectra, lower beam current, etc.)

When the crystal is { about 200 nm in size, I always measure it using a map, with drift correction. Even for larger crystals, I often still use maps to quantify thickness variations, overlapping phases and chemical gradients. If it takes 20 minutes to get enough counts for your spectrum, then a 1 hour map covering 3x the area of your desired spectrum is sufficient to get the same counts. Then you can correct for nearby materials, hole spectrum, geometry, etc.

I use homebrew software which applies k-factors and a thickness correction. I can tell it a thickness that I've measured using STXM or CBED or EELS, or if I can constrain it to a specific mineral system, then it will vary the thickness to optimize the stoichiometry (only possible with some minerals -- long discussion in that). It turns out that even 100 nm thick specimens can have significant corrections for thickness -- especially with small takeoff angles and elements { Si. So don't ignore thickness corrections, they can be very significant. Again, a map is sometimes necessary to get good estimations of actual absorption path lengths for the photons.

When the crystal is on the order of 100 nm thick, you want to make sure you are off a zone axis. This is mostly a problem with crystals that are at fixed orientation to the substrate. When you are on a zone axis, and the thickness of the crystal is on the order of an extinction length for one or another of the diffraction g-vectors, then you can get the ALCHEMI effect which will alter your EDS by a few %. In practice this rarely happens -- you have to get the alignment with a zone axis to be VERY fortuitous (or un-fortuitous), so you won't probably see it. Until one day you have a spectrum from left field that makes no sense and you can't reproduce it (because you're not on the zone axis anymore...)

Cheers,

Zack Gainsforth
Space Sciences Laboratory, UC Berkeley
7 Gauss Way
Berkeley, CA 94720
510-642-9733
zackg-at-ssl.berkeley.edu

On Mar 14, 2013, at 6:15 AM, dsherman-at-purdue.edu wrote:

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Hi All,

Have you found significant inaccuracies with X-ray mapping when using a
very high end FE-TEM (such as FEI Titan) and tilting the sample to the
angle required for maximum X-ray counts? To my knowledge, the position of
many EDS collimators requires a tilt
of 20-30 degrees in order to obtain a sufficient count to offset
background. The tilting does mean that the beam is traveling through a
thicker and angled area of the sample. My concern would be in situations
like mapping locations of molecules coating nano
particles or other instances where very accurate location of elements
relative to other nano features is desired.

Does the tilting to this amount cause sufficient offset and location error
to be a concern with very high resolution instruments?
How do you handle such situations if you do think it makes a significant
difference?
Are there some preferred detector/microscope criteria or combinations that
reduce this problem and should be considered when purchasing an EDS system
for this level of instrument? (vendor comments appreciated but use
discretion as to whether they should
be off line).

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Name: Andrew Minor

Organization: UC Berkeley & LBL

Title-Subject: [Filtered] 4th International Workshop on Remote Electron
Microscopy and In Situ Studies

Message: Dear Colleagues,

We are happy to announce the 4th International Workshop on Remote
Electron Microscopy and In Situ Studies, which will be held in Lisbon,
Portugal during May 22-24, 2013.

Building upon three previous and successful workshops (Stanford,
USA-April 2008, Göteborg, Sweden -November 2009 and Carnegie Mellon,
USA June 2011), the scope of the upcoming workshop will be to review
the state-of-the-art in remote electron microscopy, including the use of
remote approaches for education, as well as the state-of-the-art of
in-situ experiments, in particular those experiments that have enabled
major advancements in the areas of nanoscience and nanotechnology. The
event brings together around 75 people from academics and industry with
experience in electron imaging, electron spectroscopy, nanomaterials,
electron optics, stage design and fabrication, as well as recording
media and internet technology.

For registration and further information, please visit
http://www.me.utexas.edu/~insitumicroscopy2013/index.html
For questions please contact one of the organizers listed on the webpage.

Sincerely

Paulo Ferreira
Associate Professor
Materials Science and Engineering
University of Texas at Austin, USA

Andrew Minor
Associate Professor
Materials Science and Engineering
University of California at Berkeley, USA

Patricia Carvalho
Assistant Professor
Instituto Superior Tecnico
Lisbon, Portugal

Jose Maria Albuquerque
Advisor
Institute of Welding and Quality
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Email: steve.buckingham-at-novelis.com
Name: Steve Buckingham

Organization: Novelis

Title-Subject: [Filtered] Cost of analysis

Message: Hi,

I am looking for information on charges for analytical work performed by
contract labs.
We are in the process of deciding which equipment to purchase and which
work to contract out. This obviously depends on the amount of work we
have for each technique and the cost to have the work performed outside.

If anyone has any information on typical contract lab costs or knows of
a previous study that I might access I would appreciate it. The
techniques I am interested in would include XRF, XRD, ICP, SEM/EDX, XPS,
SIMS, etc.

Many Thanks,

Steve

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Hi Steve,

the examples of the microscopy and microanalysis fees are collected at
the MIAWiki Knowledge Network:
http://confocal-manawatu.pbworks.com/w/page/40058040/Microscopy%20Fees

That greatly depends on the particular circumstances of your lab,
- how well it is supported and subsidised by your company/university/society
- what is your marketing target. It is frequently a ratio of financial
revenue / volume of collaborative research (papers) you plan to receive
from/with your customers
- success of your lab is more political than commercial issue though:
http://confocal-manawatu.pbworks.com/w/page/63945076/How%20to%20Be%20Successful%20in%20Science
:0)

Your own findings and contribution to MIAWiki would be highly appreciated.

Cheers,
Dmitry

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}
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}
} Title-Subject: [Filtered] Cost of analysis
}
} Message: Hi,
}
} I am looking for information on charges for analytical work performed by
} contract labs.
} We are in the process of deciding which equipment to purchase and which
} work to contract out. This obviously depends on the amount of work we
} have for each technique and the cost to have the work performed outside.
}
} If anyone has any information on typical contract lab costs or knows of
} a previous study that I might access I would appreciate it. The
} techniques I am interested in would include XRF, XRD, ICP, SEM/EDX, XPS,
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Dear all,

Have any of you used aptamers (up to 100-nucleotide polymers of either RNA
or DNA) instead of antibodies in localisation work? There are a couple of
published papers, but seems that they've mostly been used on gels or in
pull-down assays and the like. Could be good but would be interested in
any feedback. Expensive to develop at first, but then possibly much
cheaper to re-order. I've ordered a couple of published ones to try.

cheers,
Roseamry

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Email: klemari-at-fzu.cz
Name: Mariana Klementova

Organization: Institute of Physics of the AS CR

Title-Subject: [Filtered] Windowed or window/less EDS detector for TEM?

Message: Dear colleagues,

we are about to purchase an EDS detector system for our TEM Philips CM120. The system will have a
Peltier-cooled SDD detector. We have a choice between a detector with a window and a windowless
detector. The dominant use of the system wil be for characterization of unknown crystalline phases.
In our lab we receive a variety of samples including light-element materials, but light elements are
not the principal subject of our research.

Now we have a dilemma. On one hand it is attractive to have the improved sensitivity of the detector
at low energies, but on the other hand we are concerned about possible contamination of the detector
without the protecting window.

Does anyone have an opinion or experience to share?

Thank you in advance,

Mariana

************************************************
Mariana Klementova
************************************************
Institute of Physics of the AS CR, v.v.i.
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Hello Steve,
Here's what ARDL charges...It's public information. Anyone can call up and get these numbers.

SEM/EDS/Dot map including line scans $300/sample
TEM imaging of powders (Carbon black, silica, etc.) $450
Photomicrography (stereoscope only) $115/hr or $60/photo

-----Original Message-----
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Sent: Friday, March 15, 2013 6:10 PM
To: Frank Karl

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Email: steve.buckingham-at-novelis.com
Name: Steve Buckingham

Organization: Novelis

Title-Subject: [Filtered] Cost of analysis

Message: Hi,

I am looking for information on charges for analytical work performed by
contract labs.
We are in the process of deciding which equipment to purchase and which
work to contract out. This obviously depends on the amount of work we
have for each technique and the cost to have the work performed outside.

If anyone has any information on typical contract lab costs or knows of
a previous study that I might access I would appreciate it. The
techniques I am interested in would include XRF, XRD, ICP, SEM/EDX, XPS,
SIMS, etc�....

Many Thanks,

Steve

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This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Email: martin.roe-at-nottingham.ac.uk
Name: Martin Roe

Organization: Nottingham University

Title-Subject: [Filtered] EDAX DX-4 pc system

Message: Dear List servers,
Would anyone out there have an old EDAX DX-4 integrated SEM EDX pc they no longer need? We
desperately need one for spares. We don't need any of the EDX boards inside as our system now uses a
stand-alone INCA system. Our EDX DX-4 is connected to a FEG ESEM XL30 that runs on Windows NT using
5.7 version microscope control s/w. We would pay for the transportation costs and any other
disbursements.
Thanks
Martin Roe
Nottingham University

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Email: dmdorazio13-at-gmail.com
Name: Dawn M. D'Orazio

Organization: iATL

Title-Subject: [Filtered] TEM-Independent Service Providers for EDS

Message: We have three almost brand new EDXA units on each of our HRTEMs. Two are LN2-less the
other is old-school with LN2 dewar. The vendor is a known, though smaller, supplier in our
industry. Their inability to respond to calls (though we have all units under service agreement)
has us concerned. Does anyone have contact information for any independent service providers for
EDS from VA to New England? Please contact me at ddorazio-at-iatl.com. Thank you!

Dawn DOrazio
Sample Manager
International Asbestos Testing Laboratories (iATL)

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NORTHERN CALIFORNIA MICROSCOPISTS --Northern California Society for
Microscopy

You are invited to our next meeting on the evening of March 21st at
Nanolab Technologies in Milpitas. The evening will include socializing,
business discussions, a light meal and scientific presentations.

The meeting will start at 6:00 at Nanolab Technologies, 1708 McCarthy
Blvd., Milpitas.
The program is as follows:

6 - 6:30: Registration and socializing
6:30: Dinner and short business meeting
7:30 - 9:00: Scientific talks and dessert

* Michael Isaacson, PhD.University of California at Santa Cruz:
*"Microscopy Through the Centuries: An Exploration of Microscopic
Imaging without Lenses"*
* Danielle Jorgens, Lawrence Berkeley National Laboratory and Oregon
Health & Science University: *"Seeing cells in 3D: a tale of
multi-modal imaging"*
* Ailey Crow, Genentech Inc.: *"Angiogenesis: Imaging & Image Analysis"*

--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, Diabetes & Endocrinology Research Center Microscopy Core
UCSF, Dept. of Anatomy, Rm S657
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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Greetings All,

The Midwest Microscopy and Microanalysis Society will hold its first meeting of 2013 on Friday, March 29th, at Northwestern University in Evanston, IL. The program is titled, "To Infinity and Beyond: Multidimensional Microscopy", and Bill Russin has put together an exciting lineup of presentations on 3D imaging, ultrafast electron microscopy, and other new developments in biological EM. A schedule and registration details can be found on our website under Meetings:

www.midwestmicroscopy.org

Please join us for an informative program and a chance to meet with your local colleagues and vendors. We look forward to seeing you there!

Elaine Schumacher
M3S Program Coordinator

*****************************************************************
Elaine F. Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com

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Dear colleagues,
�
I am plan to investigate the presence of micro and submicro aluminosilicate particles in rat organs.
No labeling of the particles is possible, so I would rely solely on the optical detection of the particles (micro size) or on the analytical detection of Si/Al (submicro).
The particles are not spherical, they don't have a definite shape so I expect the submicro sized particles to be hard to detect in LM.
I wondered if there would be some clever mind who could think of a�method to detect Al/Si in�histological slides (not stained,�for this purpose).
Perhaps I could simply put the slide in a SEM and do an EDX-mapping but it fear the charging (glass slide) and it would be quite time-intensive.
To avoid complications I would like to be able to detect the particles directly on a histological section placed on a glass slide in the usual way.
I know no "histological" labeling of Al/Si based on chemistry but if someone knows better, I would be glad to hear about the method.
Many thanks in advance.
�
Stephane

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Dear all,

we are looking for a second-hand cooling holder compatible with Philips CM120 for a reasonable price. Any offers?

Have a nice day,

Mariana

************************************************
�Mariana Klementova
************************************************
�Institute of Physics of the AS CR, v.v.i.
�Cukrovarnicka 10
�CZ-162 00 Praha 6
�Czech Republic
************************************************
�e-mail: klemari-at-fzu.cz

�tel. : +420 - 220 318 470
************************************************

==============================Original Headers==============================
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Hi, Stephane,

What you want to do is to have your histologist cut thicker paraffin sections. Lay one of these sections on top of a piece of double-sided carbon adhesive tape on your SEM sample holder, and remove the paraffin with xylene. Dehydrate the sample with 100% ethanol and allow the sample to air dry, then carbon-coat it. Use your back-scattered electron detector to locate your particles. Collect your secondary electron image and backscattered electron images of the sample before doing your EDS work, since the higher beam current for the EDS work will burn your tissue. After getting your photos of the tissue, do your EDS work, and you're finished. I've done this on pathology samples, and for forensic samples in the past. The technique works quite well. This works for finding ferruginous bodies in lung, for example, or talc particles, in the case of I.V. drug abuse. You definitely don't want to work from a glass slide. The nice thing about working with the carbon tape as a background is that the low atomic number background of the carbon tape will not introduce additional x-ray peaks into your spectra, and the tape helps ground your sample. I tried to work with sections 10-15 microns thick.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: nizets2-at-yahoo.com [nizets2-at-yahoo.com]
Sent: Thursday, March 21, 2013 8:26 AM
To: Haller, Edward

Dear colleagues,

I am plan to investigate the presence of micro and submicro aluminosilicate particles in rat organs.
No labeling of the particles is possible, so I would rely solely on the optical detection of the particles (micro size) or on the analytical detection of Si/Al (submicro).
The particles are not spherical, they don't have a definite shape so I expect the submicro sized particles to be hard to detect in LM.
I wondered if there would be some clever mind who could think of a method to detect Al/Si in histological slides (not stained, for this purpose).
Perhaps I could simply put the slide in a SEM and do an EDX-mapping but it fear the charging (glass slide) and it would be quite time-intensive.
To avoid complications I would like to be able to detect the particles directly on a histological section placed on a glass slide in the usual way.
I know no "histological" labeling of Al/Si based on chemistry but if someone knows better, I would be glad to hear about the method.
Many thanks in advance.

Stephane

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Stephane,

Two possible approaches:

1) Try polarizing light microscopy on stained and unstained sections. I
suggest both as it might be easier to locate the particles on unstained
material but you may need the stained material for comparison to see their
locations relative to other features.

2) Use EDS mapping with an SEM. Charging can be handled by either carbon
coating the sample or using low vacuum (preferably with an EDS adapter on
your microscope final lens. This acts to funnel the primary beam closer
to the sample while protecting it from the low vacuum environs). To make
mapping efficient, use an SDD system that will give you significant counts
even at lower kVs (I would try 10kV initially) to get maps in acceptable
time frames.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

On 3/21/13 8:20 AM, "nizets2-at-yahoo.com" {nizets2-at-yahoo.com} wrote:

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Hi, Stephane,

In many cases, I would be able to do a direct comparison of my SEM image at low magnification with a histology photograph taken with the optical microscope at the same magnification, and find the same area in both images. In some cases, I would need to photograph the tissue still in the paraffin block and compare this image to my SEM section image. Essentially, you are working with serial sections from the same block, one stained with H&E, and one deparaffinized and carbon coated, then photographed in the SEM by secondary electrons and backscattered electrons (2 images, one showing the particles by atomic number contrast). By photographing the area of interest at progressively higher magnification, you can document the area containing the particles, and then perform EDS to determine the particle makeup.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: Stephane Nizet [nizets2-at-yahoo.com]
Sent: Thursday, March 21, 2013 9:02 AM
To: Haller, Edward

Dear colleagues,

I am plan to investigate the presence of micro and submicro aluminosilicate particles in rat organs.
No labeling of the particles is possible, so I would rely solely on the optical detection of the particles (micro size) or on the analytical detection of Si/Al (submicro).
The particles are not spherical, they don't have a definite shape so I expect the submicro sized particles to be hard to detect in LM.
I wondered if there would be some clever mind who could think of a method to detect Al/Si in histological slides (not stained, for this purpose).
Perhaps I could simply put the slide in a SEM and do an EDX-mapping but it fear the charging (glass slide) and it would be quite time-intensive.
To avoid complications I would like to be able to detect the particles directly on a histological section placed on a glass slide in the usual way.
I know no "histological" labeling of Al/Si based on chemistry but if someone knows better, I would be glad to hear about the method.
Many thanks in advance.

Stephane

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Ed,

Great idea to mount sections on carbon tape. However, couldn't you do the
same by mounting on plastic coverslips (thus eliminating the Si from the
glass slides) and the be able to do SEM and LM on the same sections? If
low vacuum was used without the carbon coating than it may be possible to
stain the tissue after the SEM analysis, although I think prior staining
might be okay as well depending on the stain. Haven't tried it so not
sure on that.

A main concern would be relocation of the particles during sample prep.
Thicker sections (either thick paraffin or vibratome sections) might
reduce this possibility and would certainly be easier to work with and
allow for excising high probability areas for TEM prep and analysis.

Debby

On 3/21/13 8:54 AM, "ehaller-at-health.usf.edu" {ehaller-at-health.usf.edu}
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Hi, Debby,

I had considered your idea, but don't think this would gain anything over using a glass substrate. I didn't, and still do not, have access to an LV SEM. Plastic coverslips will usually involve the use of some type of coating to hold the sections in place during de-paraffinization/dehydration, adding another step, and the plastic, being a non-conductor, will add to your problems with charging. Some plastic coverslips are pretty beam sensitive, and you can burn pits in them with the electron beam while collecting your EDS spectrum. All of these things can factor in to your sample prep choices. Your suggestion is feasible, but I don't think you will gain much over having serial sections, one H&E and one for the SEM, since these are only microns apart, essentially only a cell or two in thickness apart in the tissue.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: dsherman-at-purdue.edu [dsherman-at-purdue.edu]
Sent: Thursday, March 21, 2013 9:58 AM
To: Haller, Edward

Ed,

Great idea to mount sections on carbon tape. However, couldn't you do the
same by mounting on plastic coverslips (thus eliminating the Si from the
glass slides) and the be able to do SEM and LM on the same sections? If
low vacuum was used without the carbon coating than it may be possible to
stain the tissue after the SEM analysis, although I think prior staining
might be okay as well depending on the stain. Haven't tried it so not
sure on that.

A main concern would be relocation of the particles during sample prep.
Thicker sections (either thick paraffin or vibratome sections) might
reduce this possibility and would certainly be easier to work with and
allow for excising high probability areas for TEM prep and analysis.

Debby

On 3/21/13 8:54 AM, "ehaller-at-health.usf.edu" {ehaller-at-health.usf.edu}
wrote:

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We all love to work with our SEM but on second though perhaps it is overkill.
It seems that dark field LM would be appropriate to observe submicroparticles with enormous advantages such as no special preparation needed and direct comparison with histology possible.
http://www.ncbi.nlm.nih.gov/pubmed/16290430

I don't know if the paraffin embedding and the observation of thin sections can�be an issue with this technique, though.
Thank you for the interesting ideas and discussions.
�
Regards,
Stephane

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http://www.jstor.org/stable/pdfplus/2442818.pdf?acceptTC=true

Article: Detection of Silica in Plants

EDX/EDS is possible, but remember that x-ray emissions in SEM are generated by the action of the beam on the surface. Si and Al are light elements with relatively weak/low energy x-rays. Thus, the depth in the section of the particles will be a large determinant of their capacity to be detected by this method.

Hope this helps,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

HomePage: http://cmirt.wcupa.edu (with link to instrument Scheduler)

-----Original Message-----
X-from: nizets2-at-yahoo.com [mailto:nizets2-at-yahoo.com]
Sent: Thursday, March 21, 2013 8:27 AM
To: Monson, Frederick

Dear colleagues,
�
I am plan to investigate the presence of micro and submicro aluminosilicate particles in rat organs.
No labeling of the particles is possible, so I would rely solely on the optical detection of the particles (micro size) or on the analytical detection of Si/Al (submicro).
The particles are not spherical, they don't have a definite shape so I expect the submicro sized particles to be hard to detect in LM.
I wondered if there would be some clever mind who could think of a�method to detect Al/Si in�histological slides (not stained,�for this purpose).
Perhaps I could simply put the slide in a SEM and do an EDX-mapping but it fear the charging (glass slide) and it would be quite time-intensive.
To avoid complications I would like to be able to detect the particles directly on a histological section placed on a glass slide in the usual way.
I know no "histological" labeling of Al/Si based on chemistry but if someone knows better, I would be glad to hear about the method.
Many thanks in advance.
�
Stephane

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2, 37 -- Date: Thu, 21 Mar 2013 05:18:34 -0700 (PDT) 2, 37 -- From: Stephane Nizet {nizets2-at-yahoo.com} 2, 37 -- Reply-To: Stephane Nizet {nizets2-at-yahoo.com} 2, 37 -- Subject: EDX on histological slides 2, 37 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} 2, 37 -- MIME-Version: 1.0 2, 37 -- Content-Type: text/plain; charset=iso-8859-1 2, 37 -- Content-Transfer-Encoding: 8bit 2, 37 -- X-MIME-Autoconverted: from quoted-printable to 8bit by ns.microscopy.com id r2LCIYEe025080 ==============================End of - Headers==============================

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15, 38 -- From FMonson-at-wcupa.edu Thu Mar 21 14:47:25 2013
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I want to thank everyone who responded to my question on TEM calibration and involvement in certification programs.

I have to admit I got very little response from the non-biological or material side of TEM consulting.

First my disclaimer: ARDL uses our TEM chiefly for carbon black and silica sizing. We are certified by A2LA and we would like to find a reasonable, but traceable standard to use for calibrate. We currently use a grating replica and calibrate at 4 different magnifications. Following our monthly calibration, I try to run a 203 nm nanosphere NIST traceable control, but only at the magnification we typically use.

This drives our QC guy nuts. Thus my survey.

In 2007 I asked the same kind of question. Those results can be found on the inactive, but still available MSNEO blog. I'll summarize it by saying of the 24 people responding, 14 used a grating replica and 15 used some sort of crystal. No information is available on the types of lab.

The current results are below.

I got 11 responses. The majority (9 or 82%) were from biological type labs. The remainder (2 or 18%) dealt with non-biological world or if you will, material sciences.

Of the biological TEM users 2 (22.2%) used the Mag-I-Cal standard. One (11.1%) lab used a 1500mesh grid and catalase crystals. The rest used a replica grating at some magnifications. Several responses indicated they used a computer supervised measuring system which extrapolates measurements from a set of calibrations to non-calibrated magnifications.

Of the two biological labs using Mag-I-Cal, one reserved it for only high magnification while the other used it on all magnifications.

None of the biological labs were certified by a A2la or LAB-like organization. I made the assumption, that if you didn't mention it, you weren't certified.

Surprising, to me, was not that both material science labs were certified, but they did not use Mag-I-Cal standard, using a grating replica instead. These lab tended to calibrate the TEM for the magnification used.

Again, thanks for your help.

Frank Karl

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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16, 24 -- From frank_karl-at-ardl.com Fri Mar 22 09:13:41 2013
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If you have a JSM-5300 and want parts,
then let me know, off-listserver. I have
to strip the unit for easy disposal. Hence,
you might as well get some parts.

This will happen in April/May.

regards,

- JQuinn

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X-from: dlowry-at-asu.edu ()

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Name: David Lowry

Organization: Arizona State University

Title-Subject: [Filtered] part for Balzers CPD020 unit

Message: We are looking for a replacement part for an old Balzers-Union
CPD020 critical-point dryer. The part in question is a solenoid valve
that controls CO2 flow and maintains set temperature for chamber
cooling. The OEM valve is Honeywell-Lucifer part# V52HRAB1300, 120V, 210
bar, 0.8 orifice. If anyone has information on a source for replacement
or salvage parts, please contact me. thank you-

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X-from: mark.grimson-at-ttu.edu ()

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Organization: Texas Tech University

Title-Subject: [Filtered] Electron emitter comparison powerpoint

Message: Hello, Our facility has instruments with each type of emitter (W, LaB6, CCFE, and
Schottky). We recently had a CCFE tip change, so I took the opportunity to image all of the
emitters in both LM and SEM at identical magnifications for direct comparison. The images were also
photographed both in and out of their respective caps. All of the images have been made into a
powerpoint presentation.

If anyone would like a copy of this powerpoint, please contact me and I would be happy to get the
presentation to you. It is fairly large, so Dropbox would probably be the best way to ship it.

Thanks, Mark

Mark Grimson
Manager, The Imaging Center
Texas Tech University

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Dear Colleague,

the Deadlines for submitting Abstracts and Early (reduced) Registration for the

40th Annual Meeting of the Society for Cutaneous Ultrastructure Research and the 6th Joint Meeting with the (Jap.) Society for Skin Structure Research

40th Annual Meeting of SCUR = 6th Joint Meeting with the SSSR
(http://orgs.dermis.net/scur/scur2013/content/e01home/e01home/index_ger.html)
Sunday 12 - Tuesday 14 May, 2013 SALZBURG, Austria
Congress Venue: Parkhotel Castellani, Salzburg
(http://www.hotel-castellani.com/)
(5 min. walk from/to the Old City Center)

have been extended to 15th of April 2013.

On behalf of the Local Organizing Committee as well as the Societies I would like to invite you to think over your active participation in the Meeting which combines 9 keynote presentations of renowned international Invited Speakers (see website link above) , most recent results in experimental as well as pathological-diagnostic Skin Research and Techniques used, exchange of knowledge and experience in a familiar atmosphere, offered at a distinguished location right in the middle of Europe.

The Local Organizing Committee is working now establishing a {second last} program which should be found included into the Meeting website until 4th of April 2013.

All necessary information
(Welcome, List of Invited speakers and their Lectures, Registration & Abstract Submission Forms, Hotel Accommodation - we have reserved a few rooms at a fairly reduced price-, etc.)

You will find on the updated
Meeting website -at- http://orgs.dermis.net/scur/scur2013/content/e01home/e01home/index_ger.html

Reduced Fees for Student application, also Travel Grant application for young researchers (contact the Secretary) available.

Therefore we encourage you to bring with you also MD and PhD students and early career researchers.
The Conference will serve as a platform where also young scientists can present their achievements and discuss them with top players in the field.

Thank you for your consideration,

have a great week,
and {Happy Easter} ,

with my best wishes and regards

Wolfgang MUSS, PhD

Secretary of SCUR (www.scur.org)
For the Local Organizing Committee
(Prof. Dr. H. Hintner, Prof. Dr. J. W. Bauer, Dr. W. Muss, Dr. R. Hametner)
(http://orgs.dermis.net/scur/scur2013/content/e01home/e01home/index_ger.html )
at work landline number:+43+662+ 4482 - (ext)4721 or -4720
at work DECT mobile phone: +43+662-4482-57704
(private mobile) Phone: +43+676-5 369 456
w.muss-at-salk.at (work) scursssr.salzbg2013-at-gmail.com wij.muss-at-aon.at (private)

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Is anyone using MACO Type S EM film? It's supposed to be a replacement
for Kodak 4489 TEM film, whose price is now ridiculous bordering on
outrageous.

Phil
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Email: mark.grimson-at-ttu.edu
Name: Mark Grimson

Organization: Texas Tech University

Title-Subject: [Filtered] Electron Emitter powerpoint

Message: Wow, huge response for the emitter powerpoint! Maybe I should have charged for it! Too
late now I guess!

The file is about 84Mb, so it is pretty big, but the images are 300dpi, so
Dropbox would be easiest, but I can always burn it on a CD and ship it that
way. If anyone has any suggestions about alternative ways of sending this,
it would be appreciated. I am not a heavy user of powerpoint, so I may have
overdone it on the size of the images.

I am putting the finishing touches on the presentation, so it should be
ready later today or tomorrow. Thanks you all for your interest, Mark

Mark Grimson, PhD
TTU Imaging Center
c/o Department of Biology
Texas Tech University
Box 43131
Lubbock TX 79409-3131

mark.grimson-at-ttu.edu
806-742-3722 x235 (Office)
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Email: gul417-at-mail.usask.ca
Name: Guosheng Liu

Organization: University of Saskatchewan

Title-Subject: [Filtered] Edwards S150B sputter coater: HT cable adaptor

Message: We had an Edwards S150B sputter coater which has run for over 20 years without major
problem except a single hassle: the lead tip of the HT cable connector has broken many times (as
shown in the photo in the link: http://www.usask.ca/biology/scopes/broken%20gold_coater.JPG). This
might due to the non-perfect design of the handler-assembly which continually puts force onto the
cable when opening/closing specimen chamber.

Original manufacture (http://www.edwardsvacuum.com/Products/List.aspx?r=125) does not have such part
available, but I guess this is standard HT cable connector which should be found somewhere in
electrical/electronic stores (online or local retailers). If anybody can provide guidance on what
this part is named (e.g. code on cable and/or adaptor) and where can I get, it would be extremely
helpful.

Thank you very much for your assistance.

Guosheng

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X-from: jawolff-at-mail.wsu.edu ()

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Email: jawolff-at-mail.wsu.edu
Name: John Wolff

Organization: Washington State University

Title-Subject: [Filtered] Electron Microprobe Research Associate, Washington State University

Message: The GeoAnalytical Laboratory in the School of the Environment, Washington State University,
seeks a Research Associate/Laboratory Manager for its electron microprobe facility. The Research
Associate will be expected to conduct their own research, seek extramural funding, and undertake
developmental work in microprobe analytical methods. Additional responsibilities include training
and providing technical advice to faculty and other research users, providing course instruction to
graduate students, and oversight and routine maintenance and operation of the JEOL 8500F electron
microprobe and the Siemens D500 X-ray powder diffractometer and relevant preparation facilities.
This is a full-time, 9-month position.

Please see {https://www.wsujobs.com} for details of the job posting and the application process.
Information about the School of the Environment and the Laboratory is available at
{environment.wsu.edu} and {www.sees.wsu.edu/Geolab/index.html} .

Enquiries may be addressed to John Wolff, {jawolff-at-mail.wsu.edu} , (+)509 335 2825.

Best wishes
JW
______________________
Prof. J.A. Wolff
Associate Director
School of the Environment
Washington State University
Pullman, WA 99164
Ph: 509 335 2825

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Dear Guosheng,

We have had a similar problem and I found out the cable is an RG 58 coaxial cable. I ordered it from a company that services those sputter coaters, but it came without any adaptor. i am trying to reuse the original one. I tired to find out what that adpator was but there are so many different adapters for coaxail cablews it is pretty hard to find the rigtht one when ou don't know the name.
good luck!

Virginia Soares
INESC-MN (Microsystems and Nanotechnologies)
R. Alves Redol, 9
1000-029 LISBON
Tel: +351213100300
Fax: +351213145843
www.inesc-mn.pt
www.in-nano.net

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Hi Phil,

We still have some Kodak 4489 available at what is a very low price
for the market.
We only have the 250 sheet available, see our web site or contact me
directly for pricing.

Thank you,

John Arnott

Disclaimer: Ladd research is in the market of selling Microscopy Lab
supplies, including those mentioned in this e-mail.

Ladd Research Industries
83 Holly Court
Williston, VT 05495

On-line Catalog: www.laddresearch.com

Telephone: 1-802-658-4961 (anywhere)
Toll Free 1-800-451-3406 (US)
Fax: 1-802-660-8859

e-mail: sales-at-laddresearch.com

At 04:30 PM 3/26/2013, you wrote:

} ----------------------------------------------------------------------------
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Email: henrik.kaker-at-guest.arnes.si
Name: Henrik Kaker

Organization: SEM-EDS and XRD laboratory, Metal Ravne, Slovenia

Title-Subject: [Filtered] Oil for the Jeol 35CF HV transformer

Message: Dear Colleagues,

We need the information (which type) about the oil in the HV transformer of our 32 years old JSM
35CF. Thanks.

Henrik Kaker
SEM-EDS & XRD Laboratory
Metal Ravne d.o.o.
Koroska cesta 14
SI-2390 Ravne na Koroskem
Slovenia
http://http://www2.arnes.si/~sgszmera1/

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Dear Henrik,
once I had the same problem with an old HV-Tank on a Zeiss EM9.
I went to the local power-supply company and asked for some liters of hv isolation oil they are using in transformers. It worked well.

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.assisi.de
www.zwillingsprojekt.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 27.03.13 14:45, schrieb microscopylistserver-noreply-at-microscopy.com:
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}
} Title-Subject: [Filtered] Oil for the Jeol 35CF HV transformer
}
} Message: Dear Colleagues,
}
} We need the information (which type) about the oil in the HV transformer of our 32 years old JSM
} 35CF. Thanks.
}
} Henrik Kaker
} SEM-EDS & XRD Laboratory
} Metal Ravne d.o.o.
} Koroska cesta 14
} SI-2390 Ravne na Koroskem
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==============================Original Headers==============================
7, 22 -- From stefan.diller-at-t-online.de Wed Mar 27 09:08:51 2013
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Mark, Pando is another possibility for up to 1GB.
http://www.pando.com/what
Does TTU have an ftp site that anyone can download from? Just another
thought. Your PPT does sound interesting.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

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Email: mark.grimson-at-ttu.edu
Name: Mark Grimson

Organization: Texas Tech University

Title-Subject: [Filtered] Electron Emitter powerpoint

Message: Wow, huge response for the emitter powerpoint! Maybe I should have
charged for it! Too
late now I guess!

The file is about 84Mb, so it is pretty big, but the images are 300dpi, so
Dropbox would be easiest, but I can always burn it on a CD and ship it that
way. If anyone has any suggestions about alternative ways of sending this,
it would be appreciated. I am not a heavy user of powerpoint, so I may have
overdone it on the size of the images.

I am putting the finishing touches on the presentation, so it should be
ready later today or tomorrow. Thanks you all for your interest, Mark

Mark Grimson, PhD
TTU Imaging Center
c/o Department of Biology
Texas Tech University
Box 43131
Lubbock TX 79409-3131

mark.grimson-at-ttu.edu
806-742-3722 x235 (Office)
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Are you adding oil to top off, or replacing oil? If you are
replacing, watch out- some of the older scopes have oil which is full
of PCBs, and you might have some difficulty getting rid of it
effectively and safely.

As for the replacement oil, any good transformer oil will do well.
The idea is that it provides a dielectric in the tank so that the HV
doesn't arc across any terminals, causing major problems in your
scope. If you are going to go through the trouble of opening up your
oil tank, you might want to check the condition of the small nylon
screws holding everything together inside the oil. I recently had a
problem with beam instability and arcing which I traced to a few
broken screws inside the oil tank which allowed the HV leads off of a
diode to get physically too close to a ground plate, causing arcing.
It's just a good practice to check. (You also don't want to have the
parts dry out- they will start to crack if they're out of the oil for
too long.)

As for the oil itself, there are a number of silicone based oils that
will work fine, or you could use a fluorinated hydrocarbon based oil.
I've heard of using mineral oil, but have not tried it.

In general, if there are import/export restrictions you need to deal
with, mineral oil is pretty benign and will be easy to obtain, and
from what I hear from other HV techs (Not microscopy related) it works
perfectly fine. (Context: I know people who work on high current high
voltage applications at 200+kV with higher currents than we ever see
in EMs.)

--Justin A. Kraft

On Wed, Mar 27, 2013 at 9:46 AM,
{microscopylistserver-noreply-at-microscopy.com} wrote:
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} Title-Subject: [Filtered] Oil for the Jeol 35CF HV transformer
}
} Message: Dear Colleagues,
}
} We need the information (which type) about the oil in the HV transformer of our 32 years old JSM
} 35CF. Thanks.
}
} Henrik Kaker
} SEM-EDS & XRD Laboratory
} Metal Ravne d.o.o.
} Koroska cesta 14
} SI-2390 Ravne na Koroskem
} Slovenia
} http://http://www2.arnes.si/~sgszmera1/
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9, 29 -- Subject: Re: [Microscopy] viaWWW:Oil for the Jeol 35CF HV transformer
9, 29 -- From: Justin Kraft {kraftpiano-at-gmail.com}
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Hi Henrik,

A long time ago, I used SHELL DIALA D as recommended by JEOL but I don't
know if this reference is still suplied.

--
Nicolas STEPHANT

Universit� de Nantes
Institut Jean Rouxel
Service de microscopie �lectronique � balayage et microanalyse
2 rue de la Houssini�re
BP 92208
44322 Nantes c�dex 3

T�l : 02 40 37 64 26
M�l : nicolas.stephant-at-univ-nantes.fr
Web : http://www.univ-nantes.fr/smebm

"Le monde n'existe que pour autant que nous sommes capables d'en
produire une image"

C.G Jung

microscopylistserver-noreply-at-microscopy.com a �crit :
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} Email: henrik.kaker-at-guest.arnes.si
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}
} Organization: SEM-EDS and XRD laboratory, Metal Ravne, Slovenia
}
} Title-Subject: [Filtered] Oil for the Jeol 35CF HV transformer
}
} Message: Dear Colleagues,
}
} We need the information (which type) about the oil in the HV transformer of our 32 years old JSM
} 35CF. Thanks.
}
} Henrik Kaker
} SEM-EDS & XRD Laboratory
} Metal Ravne d.o.o.
} Koroska cesta 14
} SI-2390 Ravne na Koroskem
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==============================Original Headers==============================
8, 25 -- From Nicolas.Stephant-at-univ-nantes.fr Thu Mar 28 04:14:37 2013
8, 25 -- Received: from mail.cnrs-imn.fr (mail.cnrs-imn.fr [193.52.97.14])
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8, 25 -- References: {201303271349.r2RDnDS1002995-at-ns.microscopy.com}
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8, 25 -- Content-Transfer-Encoding: 8bit
8, 25 -- X-Miltered: at b-mail with ID 515409FA.000 by Joe's j-chkmail (http : // j-chkmail dot ensmp dot fr)!
8, 25 -- X-j-chkmail-Enveloppe: 515409FA.000/194.214.57.230/smtp.cnrs-imn.fr/p-smtp.cnrs-imn.fr/ {Nicolas.Stephant-at-univ-nantes.fr}
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Confocal Applications Specialist Mid-west-LEI000623

OPCO Description
Leica Microsystems is a world leader in microscopes and scientific
instruments. Founded as a family business in the nineteenth century, the
company's history was marked by unparalleled innovation on its way to
becoming a global enterprise. It is a historically close cooperation with
the scientific community is the key to Leica Microsystems' tradition of
innovation, which draws on users' ideas and creates solutions tailored to
their requirements. At the global level, Leica Microsystems is organized
in three divisions, all of which are among the leaders in their respective
fields: the Life Science Division, Industry Division, and Medical
Division. Leica Microsystems has five manufacturing facilities in four
countries, with sales and service organizations in 20 countries. The
company is headquartered in Wetzlar, Germany Leica offers competitive
benefits including medical, dental, vision, prescription, long term care,
life insurance, STD, LTD and 401 (k).

Job Summary:
The Confocal Applications Specialist is a key member of the company's
sales team. The purpose of this position is to promote sales of the
companies' confocal product line in his/her assigned region. His/her
primary function is to execute domestic sales plans and to recommend
strategies to improve the sale of Confocal products in the territory, aid
in the implementation of these strategies in line with Marketing,
Corporate growth, profitability, and mission objectives.

KEY RESPONSIBILITIES:
.CUSTOMER SUPPORT: Schedules product seminars or training sessions in
concert with Leica Product & Sales Management in his/her assigned region.
Participates in all activities (travel in the field, attend industry
meetings, conferences, local and national exhibitions, etc.) that will
enhance the awareness, acceptance, and eventually the sales of Leica Life
Sciences Confocal products in his/her region.
.SATISFACTION: Insures that customers are satisfied! Also, must be capable
of working effectively with Sales Managers, PLT members, Customer Service
Technicians and Representatives, Engineers, Accounting, Manufacturing, and
Dealers to do whatever is necessary to satisfy customers' needs.
.SALES PERFORMANCE: He/she shall conduct analysis and performance
evaluation of all sales in his/her assigned region by use of the companies
Customer Resource Management tool (CRM). The evaluation includes sales
call activities, follow up on sales leads, attendance of trade shows,
contacts with local governments, associations, and academic institutions,
and traveling with other product specialists to improve their field sales
skills and product knowledge.
.SALES PLANS: Works with Sales Management to develop strategies designed
to increase sales of Leica Confocal products in assigned territory.
Specifically, he/she recommends product, pricing, advertising and sales
promotion strategies.
.NEW PRODUCTS: He/she has responsibility to increase sales of Leica
products through the introduction of new products in his/her assigned
region. Coordinates the generation of new product ideas or modifications,
performs analysis and preliminary screening of these ideas and recommends
the appropriate ones to the Area Sales Manager for acceptance or
rejection. Works with the Marketing Department in conducting field
evaluation tests to insure the viability of the new products in his/her
assigned territory. Coordinates all activities for the introduction of new
or modified products in his/her assigned territory.
.MARKETING INFORMATION: Generates and acts on the feedback on customer
preferences, suggestions, product performance and other information to
enhance the sale of Leica Confocal products.
.APPLICATION DEVELOPMENT: Works with customers or other experts to
generate appropriate field data and application notes or publications
which highlight product benefits and helps differentiate Leica products.

TRAVEL: 50-75%

Qualifications

Professional Experience
. Experience in working with Confocal Microscopy required
. Sales experience in selling products of a technical nature is
desirable
. In-depth knowledge of the microscopy market is also highly
desirable
. 2 years related microscopy experience in a laboratory minimum

Education: BA, BS or MS degree in physics or chemistry or a related
scientific field

KEY PERSONAL TRAITS
. Self-motivated
. Detail orientation
. Systematic analysis
. Good time management
. Interact as a team member
. Strong computer skills
. The candidate must possess excellent oral and written
communication skills with customers and management
. Strong technical, electrochemical, optical and mechanical aptitude
. Highly organized
. Multitasker, efficient with managing multiple accounts within a
large geographical region
. Strong passion for sales and marketing creativity
. Individual must be able to work with minimal supervision out of a
home office
. Good decision making and negotiation skills are necessary

To apply, please follow or copy and paste this link:
http://tinyurl.com/Leica-LEI000623

==============================Original Headers==============================
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12, 29 -- Message-ID: {0698544e715ee7321b27b486c3007961-at-mail.gmail.com}
12, 29 -- Subject: Leica Microsystems Confocal Specialist Mid-West
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Confocal Applications Specialist Northeast region-LEI000572

OPCO Description
Leica Microsystems is a world leader in microscopes and scientific
instruments. Founded as a family business in the nineteenth century, the
company's history was marked by unparalleled innovation on its way to
becoming a global enterprise. It is a historically close cooperation with
the scientific community is the key to Leica Microsystems' tradition of
innovation, which draws on users' ideas and creates solutions tailored to
their requirements. At the global level, Leica Microsystems is organized
in three divisions, all of which are among the leaders in their respective
fields: the Life Science Division, Industry Division, and Medical
Division. Leica Microsystems has five manufacturing facilities in four
countries, with sales and service organizations in 20 countries. The
company is headquartered in Wetzlar, Germany Leica offers competitive
benefits including medical, dental, vision, prescription, long term care,
life insurance, STD, LTD and 401 (k).

Description
Job Summary:
The Confocal Applications Specialist is a key member of the company's
sales team. The purpose of this position is to promote sales of the
companies' confocal product line in his/her assigned region. His/her
primary function is to execute domestic sales plans and to recommend
strategies to improve the sale of Confocal products in the territory, aid
in the implementation of these strategies in line with Marketing,
Corporate growth, profitability, and mission objectives.

KEY RESPONSIBILITIES:
.CUSTOMER SUPPORT: Schedules product seminars or training sessions in
concert with Leica Product & Sales Management in his/her assigned region.
Participates in all activities (travel in the field, attend industry
meetings, conferences, local and national exhibitions, etc.) that will
enhance the awareness, acceptance, and eventually the sales of Leica Life
Sciences Confocal products in his/her region.
.SATISFACTION: Insures that customers are satisfied! Also, must be capable
of working effectively with Sales Managers, PLT members, Customer Service
Technicians and Representatives, Engineers, Accounting, Manufacturing, and
Dealers to do whatever is necessary to satisfy customers' needs.
.SALES PERFORMANCE: He/she shall conduct analysis and performance
evaluation of all sales in his/her assigned region by use of the companies
Customer Resource Management tool (CRM). The evaluation includes sales
call activities, follow up on sales leads, attendance of trade shows,
contacts with local governments, associations, and academic institutions,
and traveling with other product specialists to improve their field sales
skills and product knowledge.
.SALES PLANS: Works with Sales Management to develop strategies designed
to increase sales of Leica Confocal products in assigned territory.
Specifically, he/she recommends product, pricing, advertising and sales
promotion strategies.
.NEW PRODUCTS: He/she has responsibility to increase sales of Leica
products through the introduction of new products in his/her assigned
region. Coordinates the generation of new product ideas or modifications,
performs analysis and preliminary screening of these ideas and recommends
the appropriate ones to the Area Sales Manager for acceptance or
rejection. Works with the Marketing Department in conducting field
evaluation tests to insure the viability of the new products in his/her
assigned territory. Coordinates all activities for the introduction of new
or modified products in his/her assigned territory.
.MARKETING INFORMATION: Generates and acts on the feedback on customer
preferences, suggestions, product performance and other information to
enhance the sale of Leica Confocal products.
.APPLICATION DEVELOPMENT: Works with customers or other experts to
generate appropriate field data and application notes or publications
which highlight product benefits and helps differentiate Leica products.

TRAVEL: 50-75%

Qualifications
Professional Experience
. Experience in working with Confocal Microscopy required
. Sales experience in selling products of a technical nature is
desirable
. In-depth knowledge of the microscopy market is also highly
desirable
. 2 years related microscopy experience in a laboratory minimum
Education: BA, BS or MS degree in physics or chemistry or a related
scientific field

KEY PERSONAL TRAITS
. Self-motivated
. Detail orientation
. Systematic analysis
. Good time management
. Interact as a team member
. Strong computer skills
. The candidate must possess excellent oral and written
communication skills with customers and management
. Strong technical, electrochemical, optical and mechanical aptitude
. Highly organized
. Multitasker, efficient with managing multiple accounts within a
large geographical region
. Strong passion for sales and marketing creativity
. Individual must be able to work with minimal supervision out of a
home office
. Good decision making and negotiation skills are necessary

Danaher Overview
Danaher Corporation is one of the most remarkable success stories in the
Fortune 500. In addition to Leica, Danaher owns some of the world's
leading industrial brands - products that span the most demanding
applications in the world. Currently over 50 companies and 60,000
employees around the globe are working together challenging each other to
take this success even further. Our joint quest does not only increase
chance of success, it also fosters an environment in which a global career
path is encouraged and limited only by your talent, capacity and
ambitions.

Organization
: Leica
Job Function
: Sales
Primary Location
: North America-United States-MA-Boston
Schedule
: Full-time

To apply, please follow this link: http://tinyurl.com/Leica-LEI000572

==============================Original Headers==============================
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11, 29 -- Message-ID: {fdd9a2877b2594a670f43985cedd5439-at-mail.gmail.com}
11, 29 -- Subject: Leica Microsystems Confocal Specialist Northeast
11, 29 -- To: Microscopy-at-microscopy.com
11, 29 -- Content-Type: text/plain; charset=ISO-8859-1
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Confocal Applications Specialist West - CA, AZ, NM, TX-LEI000573

OPCO Description
Leica Microsystems is a world leader in microscopes and scientific
instruments. Founded as a family business in the nineteenth century, the
company's history was marked by unparalleled innovation on its way to
becoming a global enterprise. It is a historically close cooperation with
the scientific community is the key to Leica Microsystems' tradition of
innovation, which draws on users' ideas and creates solutions tailored to
their requirements. At the global level, Leica Microsystems is organized
in three divisions, all of which are among the leaders in their respective
fields: the Life Science Division, Industry Division, and Medical
Division. Leica Microsystems has five manufacturing facilities in four
countries, with sales and service organizations in 20 countries. The
company is headquartered in Wetzlar, Germany Leica offers competitive
benefits including medical, dental, vision, prescription, long term care,
life insurance, STD, LTD and 401 (k).

Job Summary:
The Confocal Applications Specialist is a key member of the company's
sales team. The purpose of this position is to promote sales of the
companies' confocal product line in his/her assigned region. His/her
primary function is to execute domestic sales plans and to recommend
strategies to improve the sale of Confocal products in the territory, aid
in the implementation of these strategies in line with Marketing,
Corporate growth, profitability, and mission objectives.

KEY RESPONSIBILITIES:
.CUSTOMER SUPPORT: Schedules product seminars or training sessions in
concert with Leica Product & Sales Management in his/her assigned region.
Participates in all activities (travel in the field, attend industry
meetings, conferences, local and national exhibitions, etc.) that will
enhance the awareness, acceptance, and eventually the sales of Leica Life
Sciences Confocal products in his/her region.
.SATISFACTION: Insures that customers are satisfied! Also, must be capable
of working effectively with Sales Managers, PLT members, Customer Service
Technicians and Representatives, Engineers, Accounting, Manufacturing, and
Dealers to do whatever is necessary to satisfy customers' needs.
.SALES PERFORMANCE: He/she shall conduct analysis and performance
evaluation of all sales in his/her assigned region by use of the companies
Customer Resource Management tool (CRM). The evaluation includes sales
call activities, follow up on sales leads, attendance of trade shows,
contacts with local governments, associations, and academic institutions,
and traveling with other product specialists to improve their field sales
skills and product knowledge.
.SALES PLANS: Works with Sales Management to develop strategies designed
to increase sales of Leica Confocal products in assigned territory.
Specifically, he/she recommends product, pricing, advertising and sales
promotion strategies.
.NEW PRODUCTS: He/she has responsibility to increase sales of Leica
products through the introduction of new products in his/her assigned
region. Coordinates the generation of new product ideas or modifications,
performs analysis and preliminary screening of these ideas and recommends
the appropriate ones to the Area Sales Manager for acceptance or
rejection. Works with the Marketing Department in conducting field
evaluation tests to insure the viability of the new products in his/her
assigned territory. Coordinates all activities for the introduction of new
or modified products in his/her assigned territory.
.MARKETING INFORMATION: Generates and acts on the feedback on customer
preferences, suggestions, product performance and other information to
enhance the sale of Leica Confocal products.
.APPLICATION DEVELOPMENT: Works with customers or other experts to
generate appropriate field data and application notes or publications
which highlight product benefits and helps differentiate Leica products.

TRAVEL: 50-75%

Qualifications
Professional Experience
. Experience in working with Confocal Microscopy required
. Sales experience in selling products of a technical nature is
desirable
. In-depth knowledge of the microscopy market is also highly
desirable
. 2 years related microscopy experience in a laboratory minimum

Education: BA, BS or MS degree in physics or chemistry or a related
scientific field

KEY PERSONAL TRAITS
. Self-motivated
. Detail orientation
. Systematic analysis
. Good time management
. Interact as a team member
. Strong computer skills
. The candidate must possess excellent oral and written
communication skills with customers and management
. Strong technical, electrochemical, optical and mechanical aptitude
. Highly organized
. Multitasker, efficient with managing multiple accounts within a
large geographical region
. Strong passion for sales and marketing creativity
. Individual must be able to work with minimal supervision out of a
home office
. Good decision making and negotiation skills are necessary

Danaher Overview
Danaher Corporation is one of the most remarkable success stories in the
Fortune 500. In addition to Leica, Danaher owns some of the world's
leading industrial brands - products that span the most demanding
applications in the world. Currently over 50 companies and 60,000
employees around the globe are working together challenging each other to
take this success even further. Our joint quest does not only increase
chance of success, it also fosters an environment in which a global career
path is encouraged and limited only by your talent, capacity and
ambitions.

Organization
: Leica
Job Function
: Sales
Primary Location
: North America-United States-CA-San Diego
:
Schedule
: Full-time

To apply, please follow this link: http://tinyurl.com/Leica-LEI000573

==============================Original Headers==============================
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Email: Linda.Nikolova-at-hsc.utah.edu
Name: Linda

Organization: University of Utah

Title-Subject: [Filtered] Drosophil head high pressure freezing

Message: Hi All,
I am a EM specialist at the Core Facility and we have a graduate student who needs to perform high
pressure freezing on Drosophila head, which will be detached right before the feezing. Because I
have never done HPF on adult insect except eggs and larvae, my question is what kind of
cryoprotectant to use in this case.

Thank you and I appreciate very much your suggestions and help.

Linda Nikolova
EM Core Facility,
University of Utah,
Salt Lake City, UT

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Email: mark.grimson-at-ttu.edu
Name: Mark Grimson

Organization: Texas Tech University

Title-Subject: [Filtered] Emitter Powerpoint is now available

Message: Hello, everyone, There has been such a big response for this powerpoint, I think it is
easier provide the Link via the List.

The electron emitter powerpoint is now available in the DropBox. You do not need to have DropBox to
get the files, just go to the link below and you should be able to easily download them:

https://www.dropbox.com/sh/iosqhodas7ylf24/oL_xLjXZWC

There is a big 95 Mb version and a small 12Mb version (graciously shrunk by Lyle Gordon at
Northwestern).

If anybody has any trouble getting these files, the small 12Mb version can be sent as an attachment,
so please contact me, and I will get that to you.

Thank you all for your interest, and if you have any questions or comments about the presentation,
please feel free to contact me.

If you could shoot me a quick email to tell me you downloaded them, that would be much appreciated.

I will leave the files in the DropBox until I need the room! Thanks, Mark

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Email: CGoldsmith-at-cdc.gov
Name: Cynthia Goldsmith

Organization: Southeastern Microscopy Society

Title-Subject: [Filtered] 2013 meeting of the Southeastern Microscopy Society

Message: The 2013 meeting for the Southeastern Microscopy Society will be at the Embassy Suites Golf
Resort and Conference Center in Greenville, SC, on May 22-24! There is an exciting program planned
with 3 Invited Speakers: Jay Jerome from Vanderbilt University Medical Center, Paul Carpenter from
Washington University St Louis, and Brian J Ford, a well-known lecturer who is based at Cambridge
University, England.

In addition, there will be the Ruska Award presentations, contributed talks and posters, and two
workshops. There will also be a Vendor Social on Wednesday evening, a Banquet on Thursday evening,
and a Business Breakfast on Friday morning.

The SEMS website is ready for your registration, available at
http://southeasternmicroscopy.org/2013-meeting/.

Below are the descriptions of the workshops being held at Clemson University:

First, at the Clemson Light Imaging Facility featuring the Leica Imaging Suite, there will be a
workshop titled Sample techniques for creating high resolution images: tips and tricks for great
confocal and super resolution sample preparation", sponsored by Leica.

Second, at the Advanced Materials Research Laboratory, there will be a tour of the facility (with 3
TEMS (including a 300 kV) and 4 SEMs) and an introduction to the capabilities of the microscopes
there, sponsored by Hitachi, and a workshop titled A Semiconductor-Based Heating and Electrical
Biasing Platform for High-Resolution in situ Electron Microscopy, sponsored by Protochips.

We hope to see you in Greenville in May!

Cynthia Goldsmith
Secretary, SEMS

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I just joined this list. I am a member of a "hackerspace" also known as a "maker space" in Chicago, a non-profit organization that provides workshop space, tools, and instruction to members of the community. We have a wood shop, machine shop, blacksmith shop, MIG and TIG welders, electronics lab, CNC equipment (laser cutter, 3D printers, vinyl cutter, CNC milling), textile lab, and home brewing lab.

We recently received a donation of a Leica S430 SEM. It survived me disassembling it and moving it, and it works, at least for secondary electron imaging. (The Oxford Isis EDX and the four quadrant backscatter detector don't work, and the vacuum is only getting down to 8E-5 torr, but I will post more on that later.)

I'm very excited about this, as I don't know of many SEMs accessible to the general public. I'm interested if anyone else has done anything like this, and their experiences.

Now we also received a Technics Hummer V sputter coater and the optional carbon arc attachment. These came with no manuals. By any chance, does anyone have manuals for these, and if so, could I please get a scan? Or does anyone know where I can find this online?

Thanks,
Ryan

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I could use some ideas for the detection of hydrogen or Mg(OH)2 on the surface of Magnesium Oxide metal surface.

Its seems that Magnesium Oxide will absorb water over time and this will interfere with a welding process. So to resolve the problem a preheat step is done to drive off the water. The results indicate this is not as effective as we think. So would like to detect components before and after heat treatment to see how effective we are.

Any instrument techniques that could show us differences between treated and untreated areas would be useful.

Thanks

Roy Beavers
Southern Methodist University
rbeavers-at-smu.edu

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6, 30 -- Subject: Detection of elemental hydrogen or Mg(OH)2 phase in Magnesium Oxide
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Email: rajamaniselvam-at-gmail.com
Name: Rajamani Selvam

Organization: UAMS

Title-Subject: [Filtered] EM sample preparation on bones

Message: Hello all,
I am working on bones of mice models for TEM. Recently, I have encountered a problem with femurs of
the mice. When I did cross section (1um sections) of the femurs, I observed holes on sections and
osteocytes have fallen out. There are very very few osteocytes that are intact on the bone. So, I
decided to do frozen sectioning on the bones. The sections looks much better on frozen sections
with less holes and more intact osteocytes. I dont know the reason behind the osteocyte fallout, if
it could be a fixation or processing problem. Do any of you have an idea or suggestion of why its
happening? For EM sample preparation, the samples were postfixed with 1.3% osmium tetroxide, en
bloc stained with 3% uranyl acetate (made in water), dehydrated through alcohol series, infiltrated
with epoxy resin and embedded in epoxy resins for 60 C, 2 days.

Appreciate your response

Thanks
Rajamani

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Dear Mark,

thank you for sharing. I liked the presentation very much.
The table can now be accessed for reading and editing by MIAWiki permalink:
http://confocal-manawatu.pbworks.com/w/page/64943118/Electron%20Sources

Happy Easter!
Dmitry

Advanced Knowledge Management
for MICROSCOPY and Image Analysis
_________________________________________
Dmitry Sokolov, Ph.D.
Mobile: +64 21 063 5382
dmitry.v.sokolov-at-gmail.com

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My guess would be either of AFM techniques:
- scanning impedance
- scanning capacitance
- scanning spreading resistance or
- scanning Kelvin probe microscopy

Still will be a matter of the actual properties of the surfaces pre- and after treatment.

Happy Easter!
Dmitry
Advanced Knowledge Management
for MICROSCOPY and Image Analysis
_________________________________________
Dmitry Sokolov, Ph.D.
Mobile: +64 21 063 5382
dmitry.v.sokolov-at-gmail.com

On 29/03/2013 11:58 a.m., rbeavers-at-mail.smu.edu wrote:
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} I could use some ideas for the detection of hydrogen or Mg(OH)2 on the surface of Magnesium Oxide metal surface.
}
} Its seems that Magnesium Oxide will absorb water over time and this will interfere with a welding process. So to resolve the problem a preheat step is done to drive off the water. The results indicate this is not as effective as we think. So would like to detect components before and after heat treatment to see how effective we are.
}
} Any instrument techniques that could show us differences between treated and untreated areas would be useful.
}
} Thanks
}
} Roy Beavers
} Southern Methodist University
} rbeavers-at-smu.edu
}
}
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} 6, 30 -- Subject: Detection of elemental hydrogen or Mg(OH)2 phase in Magnesium Oxide
} 6, 30 -- metal
} 6, 30 -- Thread-Topic: Detection of elemental hydrogen or Mg(OH)2 phase in Magnesium
} 6, 30 -- Oxide metal
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On Mar 28, 2013, at 3:57 PM, rbeavers-at-mail.smu.edu wrote:

} I could use some ideas for the detection of hydrogen or Mg(OH)2 on
} the surface of Magnesium Oxide metal surface.
}
} Its seems that Magnesium Oxide will absorb water over time and this
} will interfere with a welding process. So to resolve the problem a
} preheat step is done to drive off the water. The results indicate
} this is not as effective as we think. So would like to detect
} components before and after heat treatment to see how effective we
} are.
}
} Any instrument techniques that could show us differences between
} treated and untreated areas would be useful.
}
Dear Roy,
Electron diffraction from the surface of the sample might do it,
depending on how the hydrogen or water adheres to the surface. Others
are much more familiar with technical aspects of this technique, so
perhaps one of them will also post.
Yours,
Bill

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I don't know about detecting hydrogen. It could be that infrared spectroscopy would detect a distinctive signal for the OH groups.

The O:Mg ratio is 1 for MgO and 2 for Mg(OH)2. I would think you should be able to detect that by EDS using low voltage to keep the excitation depth shallow so you see what is happening at the surface.

Warren
________________________________________
X-from: rbeavers-at-mail.smu.edu [rbeavers-at-mail.smu.edu]
Sent: Thursday, March 28, 2013 5:50 PM
To: wesaia-at-iastate.edu

I could use some ideas for the detection of hydrogen or Mg(OH)2 on the surface of Magnesium Oxide metal surface.

Its seems that Magnesium Oxide will absorb water over time and this will interfere with a welding process. So to resolve the problem a preheat step is done to drive off the water. The results indicate this is not as effective as we think. So would like to detect components before and after heat treatment to see how effective we are.

Any instrument techniques that could show us differences between treated and untreated areas would be useful.

Thanks

Roy Beavers
Southern Methodist University
rbeavers-at-smu.edu

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6, 30 -- Subject: Detection of elemental hydrogen or Mg(OH)2 phase in Magnesium Oxide
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Dear Listers,

Is there a preferred fixative for the preservation of the filaments that comprise the cytoskeleton? Are there any tecniques or procedures for improving the visualization of the filaments? Here at UNMC I have recently had two unrelated projects arrive where part of the goal is to see the cytoskeleton filaments. In one, the tissue is cartilage growth plate chondrocyte cells from mice, and in the other it is corpus luteum cells from bovine ovaries. There may be a third project in the future. The past few years the fad has been autophagosomes, I don't know if this is some new interest of the moment. Anyway, since I'm just a one person core facility I would appreciate all possible advice. Thanks to all for your past help and I look forward to your future help and advice.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
tbargar-at-unmc.edu
402-559-7347

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Hi, Tom,

Usually, the standard fixation in 2.5% glutaraldehyde with 0.1M phosphate buffer followed by 1% osmium tetroxide will work fine. A secret will be to cut your sections a little thicker than usual. For seeing cytoskeletal fibers, I like to cut dark gold sections. This holds true for following E.R. and vesicles as well. You will be able to follow them more easily in thicker sections. Use epoxy rather than LR White or Lowicryls, as the added density of the medium will give you better images. Also, stay away from Spurr's, again, you will not like the images. I like a combination of Ladd's LX 112 with DDSA, NMA and DMP-30 as follows:

LX 112 epoxy resin 47mL

Specially distilled DDSA 25mL

NMA (Nadic Methyl Anhydride) 28mL

Mix these three components for 1 minute by hand in a plastic specimen cup, add a magnetic stir bar, cover the cup and stir at high speed for at least 30 minutes. Do not spin fast enough to make air bubbles in the mix! After stirring this mix, add:

DMP-30 1.25mL

Use a disposable plastic pipette to add the DMP-30 to the mix. Stir the DMP-30 activator thoroughly into the embedding mix by hand, then use the magnetic stirrer to stir the complete mix for at least 30 minutes more. Each routine E.M. sample requires about 24mL of complete medium. More difficult to infiltrate samples require more embedding mix changes, and longer infiltration. Use propylene oxide as your intermediate fluid between your dehydrating fluid and your embedding mix. You can consider using reduced osmium (1% osmium tetroxide containing 1.5% potassium ferricyanide) in place of the 1% osmium tetroxide in your fixation procedure for added contrast. Mix these components immediately before using, protect this mix from room light, and rinse your tissue well with water after fixation and before dehydration. You can also en bloc stain with uranyl acetate. Following water rinsing after osmication, en bloc stain with 1.5% uranyl acetate in 0.1M sodium acetate buffer, pH 6.3, in the dark at 4��C, for 1��-2 hours. Follow this with thorough water rinsing before dehydration.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: tbargar-at-unmc.edu [tbargar-at-unmc.edu]
Sent: Monday, April 01, 2013 11:11 AM
To: Haller, Edward

Dear Listers,

Is there a preferred fixative for the preservation of the filaments that comprise the cytoskeleton? Are there any tecniques or procedures for improving the visualization of the filaments? Here at UNMC I have recently had two unrelated projects arrive where part of the goal is to see the cytoskeleton filaments. In one, the tissue is cartilage growth plate chondrocyte cells from mice, and in the other it is corpus luteum cells from bovine ovaries. There may be a third project in the future. The past few years the fad has been autophagosomes, I don't know if this is some new interest of the moment. Anyway, since I'm just a one person core facility I would appreciate all possible advice. Thanks to all for your past help and I look forward to your future help and advice.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
tbargar-at-unmc.edu
402-559-7347

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Dear Collective,

I have no experience with perfusion fixation, so forgive this if it's a stupid question: will the presence of heparin in the buffer used to clear blood cause any problems with fixation or subsequent immunolabeling? This will involve brain tissue.

Thanks!

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211

Phone: 573-884-5383
Email: tindallr-at-missouri.edu
Day job: www.emc.missouri.edu
Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
Blog: http://nadiasyard.com

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Extremely unlikely to have an effect in my opinion. Heparin is a highly charged, sulfated glycosaminoglycan so I suppose the increase number of negative charges could increase non-specific background. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: tindallr-at-missouri.edu [mailto:tindallr-at-missouri.edu]
Sent: Monday, April 01, 2013 4:02 PM
To: Phillips, Thomas E.

Dear Collective,

I have no experience with perfusion fixation, so forgive this if it's a stupid question: will the presence of heparin in the buffer used to clear blood cause any problems with fixation or subsequent immunolabeling? This will involve brain tissue.

Thanks!

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211

Phone: 573-884-5383
Email: tindallr-at-missouri.edu
Day job: www.emc.missouri.edu
Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
Blog: http://nadiasyard.com

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I guess it gets down to what you mean by filament. Microtubules are sensitive to temperature - low temperatures cause them to depolymerize so it is important to fix at room temp at least. Lots of fixatives include calcium to stabilize membranes but I seem to remember some papers suggesting that calcium was also detrimental to microtubule stability but I am less sure on that one. Intermediate filaments generally survive a lot so are probably less temperamental.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
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Sent: Monday, April 01, 2013 10:03 AM
To: Phillips, Thomas E.

Dear Listers,

Is there a preferred fixative for the preservation of the filaments that comprise the cytoskeleton? Are there any tecniques or procedures for improving the visualization of the filaments? Here at UNMC I have recently had two unrelated projects arrive where part of the goal is to see the cytoskeleton filaments. In one, the tissue is cartilage growth plate chondrocyte cells from mice, and in the other it is corpus luteum cells from bovine ovaries. There may be a third project in the future. The past few years the fad has been autophagosomes, I don't know if this is some new interest of the moment. Anyway, since I'm just a one person core facility I would appreciate all possible advice. Thanks to all for your past help and I look forward to your future help and advice.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
tbargar-at-unmc.edu
402-559-7347

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Hello Randy and Tom,

I don't think heparin will be an issue. Unless �. the primary antibody would be directed against a component that is heparin-like, then you might expect some labeling of the blood vessel walls. Heparin's negative charge at physiological pH is not likely to give non-specific background as any heparin left behind after perfusion would repulse negatively charged gold conjugates.

Jan

Jan Leunissen
from a brilliant summer in Dunedin, NZ

i: www.aurion.nl

On 2/04/2013, at 11:48 AM, PhillipsT-at-missouri.edu wrote:

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} Extremely unlikely to have an effect in my opinion. Heparin is a highly charged, sulfated glycosaminoglycan so I suppose the increase number of negative charges could increase non-specific background. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://biology.missouri.edu/people/?person=88
} http://www.biotech.missouri.edu/mcc/
}
}
} -----Original Message-----
} X-from: tindallr-at-missouri.edu [mailto:tindallr-at-missouri.edu]
} Sent: Monday, April 01, 2013 4:02 PM
} To: Phillips, Thomas E.
} Subject: [Microscopy] TEM/Immuno: Heparin and fixation
}
}
}
}
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} Dear Collective,
}
} I have no experience with perfusion fixation, so forgive this if it's a stupid question: will the presence of heparin in the buffer used to clear blood cause any problems with fixation or subsequent immunolabeling? This will involve brain tissue.
}
} Thanks!
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility
} W122 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
}
} Phone: 573-884-5383
} Email: tindallr-at-missouri.edu
} Day job: www.emc.missouri.edu
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} 10, 31 -- "White, Tommi A."
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} 21, 32 -- From: "Phillips, Thomas E." {PhillipsT-at-missouri.edu}
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Email: vakimler-at-mi.rr.com
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Cryoultramicrotomy of Brown Adipose Tissue

Message: Does anyone out there have any good suggestions for managing tissue integrity and
immunolabeling of brown adipose tissue (and eventually then white adipose tissue) using
cryoultramicrotomy for IVTEM (200kV)?
1) temperature of cutting? (diamond or glass knife and temperature)?
2) chamber or chuck temperature?
3) section thickness?
4) speed of sectioning?
5) immunogenicity? Fluoronanogold - or immunogold - conjugated secondary antibodies - which is best
for lipid droplet proteins?
Any advice would be most helpful. It is our first time working with brown adipose tissue in this
system. Our tissue integrity for heart is not bad at all for starters.
Thanks so much,
Vickie

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Email: mark.grimson-at-ttu.edu
Name: Mark Grimson

Organization: Texas Tech University

Title-Subject: [Filtered] Addition to Emitter PowerPoint on WWW link

Message: Hello, Debby Sherman asked for information concerning the instruments that the emitter
images were derived from. We made a new slide that fits well between slides 2 and 3 in the original
'electron emitter' powerpoint ( called 'emitter identification'). Please feel free to add this
slide to the powerpoint, and I will also add it to the copies in the dropbox.

Someone also asked if SEM images of the same sample from each piece of equipment could be added for
direct comparison. I have a W , Schottky, and CCFE, but no LaB6 SEM. If anyone has an idea for an
appropriate comparative sample (i.e with suitable detail for the CCFE), I could do 3 of the 4
instruments , but will need to find someone with a LaB6 SEM to do that one.

Again, the link to the site is:

https://www.dropbox.com/sh/iosqhodas7ylf24/oL_xLjXZWC

Thanks, Mark

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Microtubules are, indeed, sensitive to cold, so fix at room temp. I used
to have to decalcify crustacean skeleton with EDTA in with my fixative,
and the microtubules came out looking great! Other filaments are more
stable, although sometimes elastin gets faint.

Aloha,
Tina

} I guess it gets down to what you mean by filament. Microtubules are sensitive to temperature - low temperatures cause them to depolymerize so it is important to fix at room temp at least. Lots of fixatives include calcium to stabilize membranes but I seem to remember some papers suggesting that calcium was also detrimental to microtubule stability but I am less sure on that one. Intermediate filaments generally survive a lot so are probably less temperamental.
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://biology.missouri.edu/people/?person=88
} http://www.biotech.missouri.edu/mcc/
}
}
} -----Original Message-----
} X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu]
} Sent: Monday, April 01, 2013 10:03 AM
} To: Phillips, Thomas E.
} Subject: [Microscopy] Best way to preserve and visualize cytoskeleton filaments
}
}
}
}
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} Dear Listers,
}
} Is there a preferred fixative for the preservation of the filaments that comprise the cytoskeleton? Are there any tecniques or procedures for improving the visualization of the filaments? Here at UNMC I have recently had two unrelated projects arrive where part of the goal is to see the cytoskeleton filaments. In one, the tissue is cartilage growth plate chondrocyte cells from mice, and in the other it is corpus luteum cells from bovine ovaries. There may be a third project in the future. The past few years the fad has been autophagosomes, I don't know if this is some new interest of the moment. Anyway, since I'm just a one person core facility I would appreciate all possible advice. Thanks to all for your past help and I look forward to your future help and advice.
}
} Tom Bargar
} UNMC
} Electron Microscopy Core Facility
} 986395 Nebraska Medical Center
} Omaha, NE 68198-6395
} tbargar-at-unmc.edu
} 402-559-7347
}
}
}
} ==============================Original Headers==============================
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****************************************************************************
* 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*
****************************************************************************

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Email: rossjeta-at-gmail.com
Name: Jetaime

Organization: University of Maryland Baltimore

Title-Subject: [Filtered] Looking for a "good" paper on Electron Microscopy

Message: Hi,

I am a first year Biochemistry and Molecular Biology graduate student and I am looking for an easy
to understand paper on Electron Microscopy since I do not have much background in this technique. I
would greatly appreciate any suggestions!

Best regards,

Jetaime

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Hi everybody, I would like to bring to your attention our

May 29 and 30 Meeting "Advances in Structural and Chemical Imaging" at
CAMCOR, U of Oregon, Eugene, OR,

Zero participation fee, free coffee, bakery basket, lunch, finger food
and drinks at the poster session. We plan to have up to 25 posters in
addition to our 22 invited speakers.

For registration (and much more information), go to
http://nanocrystallography.net/ASCI2013. Posters shall also be
registered there.

------------------------------------------------------------------------------------------------------------
Advances in Structural and Chemical Imaging (ASCI 2013)

(A PREMIER* Network Conference), CAMCOR, Eugene, OR, May 29 - 30, 2013
(* Pooled Resources for Electron Microscopy Informatics Education and Research)

Day 1

Courtesy shuttle from Valley River Inn hotel to meeting venue 7:15 am,
bakery basket and coffee at 7:45 am (provided by PREMIER)

8.15-8.30 Welcome and Introduction to ASCI and the PREMIER Network �
Nigel D. Browning, PNNL

Session 1 Advances in Instrumentation I - Nigel D. Browning, PNNL (Chair)

8.30-9.00 Vortex Beams � Ben McMorran, U. Oregon
9.00-9.30 The UTEM project � Bernd Kabius, PNNL
9.30-10.00 Enabling observations of protein structural dynamics with
fast pump-probe electron microscopy and x-ray diffraction � James
Evans, PNNL
10.00-10.30 Coffee Break (provided by PREMIER)
10.30-11.00 Single molecule super-resolution microscopy for cancer
biology / Correlative Light and Electron Microscopy � Xiaolin Nan,
OHSU
11.00-11.30 Multiscale Imaging of Cells: From Organelle Systems to
Molecular Complexes � Gina Sosinsky, UC San Diego
11.30-12.00 Tomography of Catalysts � Ilke Arslan, PNNL

12.00-1.30 pm Lunch Break (provided by PREMIER)

Imaging Methods � Session 2 Dave C. Johnson, U. Oregon (Chair)

1.30-2.00 Contrast Improvement for Field Enhancement Based Near-field
Imaging � Erik S�nchez, PSU
2.00-2.30 The Debye-Waller factor of graphene � Chris Regan, UCLA
2.30-3.00 Imaging of Viruses by Cryogenic Transmission Electron
Microscopy and Three-Dimensional Reconstruction � David Belnap, U of
Utah
3.00-3.30 Coffee Break (provided by PREMIER)
3.30-4.00 Time-Resolved Electrostatic Force Microscopy of Thin Film
Solar Cells � David S. Ginger, UW
4.00-4.30 Imaging of Nanoparticle Dynamics on Oxide Surfaces � Abhaya Datye, UNM
4.30-5.00 Forisomes-Plant Derived ATP Independent Proteins Actuators �
Michael Knoblauch, WSU
5:00-5:30 Exploring Chemical and Structural Parameter Space - Thomas Vogt, USC

5.30-7.30 Poster Session

Day 2
Courtesy shuttle from Valley River Inn hotel to meeting venue 7:45 am,
bakery basket and coffee at 8:00 am (provided by PREMIER)

Session 1 Applications of Microscopy I � Erik S�nchez, PSU, (Chair)

8.30-9.00 Microstructure in nanomagnetism - open questions and
challenges � Kannan Krishnan, UW
9.00-9.30 Can you see me now? The power of analytical
aberration-corrected scanning transmission electron microscopy �
Robert F. Klie, UIC
9.30-10.00 Characterization of ODS alloys by SPS - A combinational TEM
and APT study � Yaqiao Wu, BSU
10.00-10.30 Coffee Break (provided by PREMIER)
10.30-11.00 Atom Probe - Brian Gorman, CSM
11.00-11.30 Bicrystallography for structural grain-boundary research �
Bryant York, PSU
11.30-12.00 Applied crystallography in SPM and TEM & open-access
crystallographic databases, Peter Moeck, PSU

12.00-1.30 pm Lunch Break (provided by PREMIER)

Session 2 Applications of Microscopy II � Peter Moeck, PSU (Chair)

1.30-2.00 A possible application of biomimetics in optimizing the
quantum efficiency of photovoltaics � Andreas Holzenburg, Texas A&M

2.00-2.30 Structural Analysis of Organic Perovskites and Bimetallic
Carbides for Photovoltaic Applications � Erwin M. Sabio, U. Wyoming

2.30-3.00 Diffraction Tomography & Automated Crystallite Orientation
and Phase Imaging in TEM, Amith Darbal, NanoMEGAS

3.00-3.15 Coffee Break (provided by PREMIER)

3.15-5.00 PREMIER Network Business Meeting

--
Peter Moeck, PhD (Dr. rer. nat. Crystallography)
Associate Professor of Physics

Nano-Crystallography Group:
http://nanocrystallography.research.pdx.edu/nano-crystallography-group.
Free (as in free speech rather than free beer) crystallographic information
and interactive 3D visualizations of crystal structures and morphologies
with educational relevance:
http://nanocrystallography.research.pdx.edu/databases.

Portland State University
Department of Physics
P.O. Box 751, Portland, OR 97207-0751
Tel.: 503 725 4227, Departmental Fax.: 503 725 2815
pmoeck-at-pdx.edu
www.physics.pdx.edu/~pmoeck

North American mirror of the Crystallography Open Database, COD,
http://nanocrystallography.org, with more than 210,000 carefully evaluated
datasets for download and interactive visualization in 3D, for more
interactive 3D display options try:
http://nanocrystallography.research.pdx.edu/search/codmirror/
Webportal: Open Access Crystallography Resource Portal
http://nanocrystallography.net.

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A local University seeks an older EDS system with or without a Detector to replace their obsolete Polaroid imaging system. If you know of a system that may be available please respond. Thanks.

Jeff Day
Dallas Texas Area

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Get 32GB Apple iPad for as low as $30.93. Limit 1.Day. Grab yours Now!
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Hello Jetaime,
I don't know of a paper but the best reference I know for beginning electron
microscopy is the book by John Bozzola and Lonnie Russell titled "Electron
Microscopy Principles and Techniques for Biologists". It's a treasure of
information beginning with a brief history of EM. The book covers specimen
preparation for TEM and SEM, optics and theory of the electron microscope
and much more. It's a volume I recommend to all beginning EM students. Good
luck and happy "EMing"

Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

X-from: rossjeta-at-gmail.com ()

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Email: rossjeta-at-gmail.com
Name: Jetaime

Organization: University of Maryland Baltimore

Title-Subject: [Filtered] Looking for a "good" paper on Electron Microscopy

Message: Hi,

I am a first year Biochemistry and Molecular Biology graduate student and I
am looking for an easy
to understand paper on Electron Microscopy since I do not have much
background in this technique. I
would greatly appreciate any suggestions!

Best regards,

Jetaime

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============================================

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Try hunting down a copy (in PDF) of:

Introduction to the
Scanning Electron Microscope
Theory, Practice, & Procedures

Prepared by
Michael Dunlap

Presented by the
FACILITY FOR ADVANCED INSTRUMENTATION,
U. C. Davis
1997

52 Page concise overview for SEM.

Tony
.........................................
Andrew Anthony "Tony" Havics, CHMM, CIH, PE
Environmental, Health & Safety, Microscopy, & Forensic Engineering
pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
(317) 718-7020 Office
(317) 718-7038 Fax
(317) 409-3238 Cell
aahavics-at-pH2LLC.com
www.ph2LLC.com

-----Original Message-----
X-from: pekysar-at-ucdavis.edu [mailto:pekysar-at-ucdavis.edu]
Sent: Wednesday, April 03, 2013 1:12 PM
To: ph2-at-sprynet.com

Hello Jetaime,
I don't know of a paper but the best reference I know for beginning electron
microscopy is the book by John Bozzola and Lonnie Russell titled "Electron
Microscopy Principles and Techniques for Biologists". It's a treasure of
information beginning with a brief history of EM. The book covers specimen
preparation for TEM and SEM, optics and theory of the electron microscope
and much more. It's a volume I recommend to all beginning EM students. Good
luck and happy "EMing"

Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

X-from: rossjeta-at-gmail.com ()

This Question/Comment was submitted to the Microscopy Listserver
using the WWW based Form at
http://microscopy.com/MicroscopyListserver/MLFormMail.html
---------------------------------------------------------------------------
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---------------------------------------------------------------------------

Email: rossjeta-at-gmail.com
Name: Jetaime

Organization: University of Maryland Baltimore

Title-Subject: [Filtered] Looking for a "good" paper on Electron Microscopy

Message: Hi,

I am a first year Biochemistry and Molecular Biology graduate student and I
am looking for an easy
to understand paper on Electron Microscopy since I do not have much
background in this technique. I
would greatly appreciate any suggestions!

Best regards,

Jetaime

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---------------------------------------------------------------------------

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Jetaime -

One of the major EM manufacturers has produced a good short article:
http://www.fei.com/resources/student-learning/introduction-to-electron-microscopy/intro.aspx

You will find lots of information on the various types of light
microscopy at http://microscopy.fsu.edu

Caroline

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Email: tnkor6ie-at-vt.edu
Name: Thess Zu

Organization: Virginia Tech

Title-Subject: [Filtered] Quantification guide for Raman Spectroscopy

Message: Dear all,

I am currently working with some biological samples trying to deduce possible physiological changes
with treatments.

I need help with resource on how to effectively quantify my data to be able to make my deductions.

Any suggestions on how to quantify my data will be very helpful

best regards,
Theresah

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Dear Listers,

I've recently experienced a contamination on my sections that doesn't look like the typical Uranyl acetate or Lead Citrate precipitates. Rather it looks like large thin flake like particles. I use disposable .22 micron filters that fit on a syringe for filtering my stains. I'm assuming it is possible to get contaminate particles from filters. I would like to hear from anyone who has dealt with contaminants from filters. I can't attached a photo to this request, but contact me by regular e-mail and I can share an image if that would help in determining the problem. Thanks

Tom Bargar
UNMC
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

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Hi Tom,
There are so many places your contamination can originate from! Have you
tried looking at your sections without any stain to check if the
contamination is present BEFORE the staining process? I would be good to
rule out that particular problem.
You can inherit contamination from your boat water, eyelash manipulator,
forceps and just from any dusty air circulating. When I see detritus on my
sections which doesn't look like stain contaminate, I usually thoroughly
clean everything in my microtome room starting with my forceps, manipulator
soaking my diamond knife in DDH2O before cleaning it.
Good luck! We've all been there!

Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

-----Original Message-----
X-from: tbargar-at-unmc.edu [mailto:tbargar-at-unmc.edu]
Sent: Thursday, April 04, 2013 8:11 AM
To: pekysar-at-ucdavis.edu

Dear Listers,

I've recently experienced a contamination on my sections that doesn't look
like the typical Uranyl acetate or Lead Citrate precipitates. Rather it
looks like large thin flake like particles. I use disposable .22 micron
filters that fit on a syringe for filtering my stains. I'm assuming it is
possible to get contaminate particles from filters. I would like to hear
from anyone who has dealt with contaminants from filters. I can't attached
a photo to this request, but contact me by regular e-mail and I can share an
image if that would help in determining the problem. Thanks

Tom Bargar
UNMC
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

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Hi Tom,

I'll be happy to take a look at your picture. We sometimes (often) see contamination that looks like flakes----they often resemble the little things that make up the "dust" on butterfly wings, if you have ever seen something like that. I'm not sure where it comes from, but we sometimes refer to it as boat contamination. When we find it, we clean our diamond knives again, make sure our boat water is clean by changing the syringe and filters we use for filling them. Sometimes that helps. Sometimes not.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
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Dear All,
since some years I worked on a new concept of visualizing specimen in the scanning electron microscope.

My profession as a photographer leads me first to search for scientifically correct but also aesthetically appealing images,
which makes me believe, that the scientific community and with it every researcher should be committed to share the aesthetics of
the microworld with as many people as possible. In scanning electron microscopes (SEMs), data is usually shown using grayscale
images. Colours can be added easily with additional detector signals. This procedure is widely accepted for communicating results
on up-to-date research to newspapers, magazines and TV broadcasting outside the scientific world.

In the past mostly still shots of the specimen had been shown, or noisy TV rate screen captures. I thought it would be worth to
bring the effects of a good movie into the SEM: fluent movement of the specimen, changing colour and lighting effects during the
sequence, chances of focus and magnification.

Find out more at my website www.nanoflight.info
For those of you who like to see immediately, what`s this all about, go to
www.nanoflight.info/nanoflight2.html
directly and hope, that the server keeps running ;-)

I am open to any discussion.

All the best,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

==============================Original Headers==============================
11, 21 -- From stefan.diller-at-t-online.de Mon Apr 8 07:35:36 2013
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Stefan

Your images look as if they use a similar multi-detector technique developed
about 20 years ago by a colleague David Scharf a US Photographer and Microscopist.

You might like to look at his work

http://www.scharfphoto.com

David has also contributed to a number of movies (iMax included) and
television specials.

Disclaimer: I have no financial interests with either of the above . I do however
appreciate the excellent images and attention to detail done by both of
these individuals.

Regards

Nestor
Your Friendly Neighborhood SysOp

Begin forwarded message:

} From: stefan.diller-at-t-online.de
} Subject: [Microscopy] A new way to visualize specimen in
} Date: April 8, 2013 7:35:36 AM CDT
} To: zaluzec-at-aaem.amc.anl.gov
} Reply-To: stefan.diller-at-t-online.de
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Dear All,
} since some years I worked on a new concept of visualizing specimen in the scanning electron microscope.
}
} My profession as a photographer leads me first to search for scientifically correct but also aesthetically appealing images,
} which makes me believe, that the scientific community and with it every researcher should be committed to share the aesthetics of
} the microworld with as many people as possible. In scanning electron microscopes (SEMs), data is usually shown using grayscale
} images. Colours can be added easily with additional detector signals. This procedure is widely accepted for communicating results
} on up-to-date research to newspapers, magazines and TV broadcasting outside the scientific world.
}
} In the past mostly still shots of the specimen had been shown, or noisy TV rate screen captures. I thought it would be worth to
} bring the effects of a good movie into the SEM: fluent movement of the specimen, changing colour and lighting effects during the
} sequence, chances of focus and magnification.
}
} Find out more at my website www.nanoflight.info
} For those of you who like to see immediately, what`s this all about, go to
} www.nanoflight.info/nanoflight2.html
} directly and hope, that the server keeps running ;-)
}
} I am open to any discussion.
}
} All the best,
} Stefan
}
}
} --
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 Phone
} ++49-931-7848701 Fax
} ++49-175-7177051 Mobile
}
} Websites:
} www.nanoflight.info
} www.stefan-diller.com
} www.electronmicroscopy.info
} www.elektronenmikroskopie.info
} www.zwillingsprojekt.de
} www.assisi.de
} Anfahrt: http://Mail.map24.com/Stefan.Diller
} -----------------------------------------------------
}
} ==============================Original Headers==============================
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} ==============================End of - Headers==============================

===========================================
Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
Materials Science/Bldg 212
9700 S. Cass Ave.
Argonne, Illinois 60439 USA
Tel: 530-NES-TORZ (530-637-8679) Fax: 630-252-4798

iChat:Zaluzec-at-AIM
Skype: Zaluzec-at-ANL
TPM: http://tpm.amc.anl.gov

Email: Zaluzec-at-aaem.amc.anl.gov
WWW: http://tpm.amc.anl.gov

Senior Scientist - Argonne National Laboratory
Fellow of the Microscopy Society of America
Senior Fellow of the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
Past-President: Microscopy Society of America
===========================================

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Dear Stefan,

thank you for the links. The images and movies are fascinating indeed.

I understand how important the appealing images are for the success in
science:
http://confocal-manawatu.pbworks.com/w/page/63945076/How%20to%20Be%20Successful%20in%20Science

However, I am doubtful about the scientific significance of the
artificially coloured images unless the colour contrast represents
variations or differences in physical / chemical properties where the
fluorescence and Raman scattering microscopy or hyperspectral imaging
for material characterisation could be the examples.

If we touch the technological base of the imaging as the source of the
primary data for scientific knowledge:
http://confocal-manawatu.pbworks.com/w/page/16347068/Technology%20of%20Research
colouring the images (unless it leads to the production of new
knowledge) is another technological operation. It adds time to the
technological sequence of image acquisition and/or analysis and
therefore reduces the efficiency of research.

Unfortunately, I have a little experience in colouring of SEM images and
I would appreciate your comments very much.

With kind regards,
Dmitry

Advanced Knowledge Management
for MICROSCOPY and Image Analysis
________________________________
Dmitry Sokolov, Ph.D.
Mob: +64 21 063 5382
dmitry.v.sokolov-at-gmail.com

On 09.04.2013 0:50, stefan.diller-at-t-online.de wrote:
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} Dear All,
} since some years I worked on a new concept of visualizing specimen in the scanning electron microscope.
}
} My profession as a photographer leads me first to search for scientifically correct but also aesthetically appealing images,
} which makes me believe, that the scientific community and with it every researcher should be committed to share the aesthetics of
} the microworld with as many people as possible. In scanning electron microscopes (SEMs), data is usually shown using grayscale
} images. Colours can be added easily with additional detector signals. This procedure is widely accepted for communicating results
} on up-to-date research to newspapers, magazines and TV broadcasting outside the scientific world.
}
} In the past mostly still shots of the specimen had been shown, or noisy TV rate screen captures. I thought it would be worth to
} bring the effects of a good movie into the SEM: fluent movement of the specimen, changing colour and lighting effects during the
} sequence, chances of focus and magnification.
}
} Find out more at my website www.nanoflight.info
} For those of you who like to see immediately, what`s this all about, go to
} www.nanoflight.info/nanoflight2.html
} directly and hope, that the server keeps running ;-)
}
} I am open to any discussion.
}
} All the best,
} Stefan
}
}

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Hi there

I think the (photoshop or similar) color on SEM micrographs once added helps
later other people, including scientists, to distinguish structures that
will take time to see without color. Nature loves camouflage; for instance
sperm tails on the surface of a human egg (zona pellucida) are very similar
to the structure of the zona and difficult to identify. Same is with
cellular projections that spread over other cells etc. It could be an option
for scientific journals to accept colored SEM micrographs if submitted along
with the originals and checked for accuracy.

Cheers

yorgos

----- Original Message -----
X-from: {dmitry.v.sokolov-at-gmail.com}
To: {eikonika-at-otenet.gr}
Sent: Monday, April 08, 2013 4:34 PM

Wow!!! Those are amazing! I just last week started dabbling in an uncolored version attempting to do similar, but I can say I certainly had a more constrained vision of where it could go. The technical challenges are considerable. My hat is off to you. Very well done.

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012�� MRC104
Washington DC 20013-7012
202-633-0891

-----Original Message-----
X-from: stefan.diller-at-t-online.de [mailto:stefan.diller-at-t-online.de]
Sent: Monday, April 08, 2013 8:37 AM
To: Whittaker, Scott

Dear All,
since some years I worked on a new concept of visualizing specimen in the scanning electron microscope.

My profession as a photographer leads me first to search for scientifically correct but also aesthetically appealing images, which makes me believe, that the scientific community and with it every researcher should be committed to share the aesthetics of the microworld with as many people as possible. In scanning electron microscopes (SEMs), data is usually shown using grayscale images. Colours can be added easily with additional detector signals. This procedure is widely accepted for communicating results on up-to-date research to newspapers, magazines and TV broadcasting outside the scientific world.

In the past mostly still shots of the specimen had been shown, or noisy TV rate screen captures. I thought it would be worth to bring the effects of a good movie into the SEM: fluent movement of the specimen, changing colour and lighting effects during the sequence, chances of focus and magnification.

Find out more at my website www.nanoflight.info For those of you who like to see immediately, what`s this all about, go to www.nanoflight.info/nanoflight2.html
directly and hope, that the server keeps running ;-)

I am open to any discussion.

All the best,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
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www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
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As I mentioned earlier on this list, I'm one of the people setting up a
donated Leica Stereoscan 430 in a hackerspace in Chicago.

We are trying to troubleshoot a problem with the backscatter detector where
we are getting no signal at all, not even noise. Any help would be greatly
appreciated. In particular, any service manuals, pinouts, and/or schematics
for the scope and the backscatter detector are very much needed.

X-from what I can gather:

According to the packing list when the scope was purchased ca. 1993-94,
it is a "430-4 BSDX (4 ELEMENT BACK SCATTER DETECTOR) part no. 760715,
also called "4 ELEMENT BSD EXTENDED" on the packing list. I can send photos
of all the parts mentioned below if that might help identify them.

I think the detector came in a box that says "K. E. Developments Limited
Backscattered Electron Detector Diode Type 7. 24 mm. Low Voltage. 1keV+
The feedthrough from the chamber has 7 pins in a round configuration.

The pre-amp box has no visible external markings, although I haven't removed
the bracket that secures it to the side of the chamber to examine it.
It has a thick cable that goes to the amp board, and connects via a DB-15
connector.

The amp board, which presumably selectively adds or inverts the four
quadrants, is attached to a metal bracket that is screwed to the power
supply in the lower left part of the desk that holds the Image Processor
and CPU boards. It says it was made by K. E. Developments Ltd. Toft,
Cambridge. This board has four connectors:

* A very large edge connector on the left. This goes into an adapter cable
that becomes a rather thin ribbon cable and connects to a bus of DB-9
connectors in the cable passage area under the back of the desk. This
appears to be a power distribution bus.

* The DB-15 input from the pre-amp, on the upper right.

* A tiny coaxial signal cable on the middle right that goes to PL-16 on the
Image Processor board.

* A 16 pin ribbon cable that goes to the Stage and Vacuum board, where it
becomes a DB-25 connector and plugs into SK58.

Troubleshooting efforts to date:

We've determined that there is definitely voltage getting to the amp
board. And I see voltages on some of the DB-15 pins going to the pre-amp.

The Image Processor port PL-16 is configured as 100 mV bipolar. I enabled
split scan to display both secondary electron and backscatter. I then
disconnected the secondary electron detector (which is 50 mV unipolar) and
connected it into PL-16. I saw a rather dark image on the backscatter side of
the screen. So I think the Image Processor input for the backscatter
detector, at least, is working, and I am operating on the hypothesis
that the amp board isn't sending it any signal. We can't see anything
coming out of the amp board via an oscilloscope either.

I'm focusing now on the 16 pin ribbon cable, which I believe has to be what
the scope uses to configure the backscatter for quadrants (add, invert, or
off) and, possibly, the four levels of gain. The board itself seems
extremely simple. I just see op amps and two hex inverters. I don't see
any microcontrollers, shift registers, or latches. So my hypothesis is that
the 16 pin ribbon cable uses logic levels to switch quadrants on or off, and
invert the channels. I'd expect 4 pins to add the quadrants and 4 pins to
subract them. Or, say, 4 pins to enable the quadrants and 4 pins to invert
them.

So we tested the ribbon cable and, regardless of quadrant setup, we don't
see any voltage on any of the pins of the ribbon cable. If this means the
board doesn't add any quadrants, this would explain what we're seeing.
I'm suspecting there's a software/config issue where the LEO software
isn't sending anything on SK58.

In the config menu, I see it configured as a 4 quadrant Mk. 1. The only
other reasonable choice would be Mk. 2, I tried that, and we still get
no signal or any voltage on any pin of the ribbon cable.

Any advice would be greatly appreciated.

Thanks,
Ryan

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Stephan, Dmitry, and Yorgos:

1. My Take - Adding color can:

A) Add to the aesthetics

B) Add contrast, and thus aid in recognition of
morphology/structure/arrangement.

C) Add spatial separation and thus aid in recognition of
morphology/structure/arrangement.

D) Add contrast or spatial separation and thus increase the speed of
recognition (efficiency).

Unfortunately, it can also distort (heighten or suppress) the underlying
mathematical correlation (distance, shape, frequency), and even distract
from the content.

Note: We already accept distortions of phase into contrast [e.g., DIC (gray
& color), PCM (gray), Hoffman (gray), etc.] or spatial position into color
contrast [staining]; so the question is really one of reliability,
relevance, reproducibility and transparent representation. There is also a
sense of naturalness that might also be important. Think about the contrast
effect of photographing a person's face with only light from the bottom up;
it is much different than light from the top (natural sunlight) down, and
thus the bottom up creates an eerie effect as a result of our human
conditioning.

2. Marlana Coe's Comments [Human Factors for Technical Communicators]
on color:

Graphics, icons, and color perform vital functions:

Adds dimension
Aids in decision making
Enhances recall
Focuses attention
Renders images more realistic
Reveals organization and pattern
Satisfies users' preferences
Speeds searches

She adds a couple of interesting comments:

Users expect color in online information, but not necessarily in hardcopy.
Hardcopy technical information is not enhanced by coloring the background or
text. User's find it tiring, confusing and distracting.

Tony

Tony's Refs:

Burnham, Color Perception in Small Test Fields, JOSA, 43, 10, 899-902, 1953
Burnham, Comparison of Color Systems with Respect to Uniform Visual Spacing,
JOSA, 39, 5, 387-391, 1949
Carter, Color and conspicuousness, JOSA, 71, 6, 723-729, 1981
Caywood, Independent Components of Color Natural Scenes Resemble V1 Neurons
in Their Spatial and Color Tuning, J Neurophysiol, 91, 2859-2873, 2004
Cote, Optical system performance visualization, Proc 1999 SPIE Annual Conf,
SPIE, 1-12, 1999
Cramb, Black and White versus Colour Photography, JFSS, 4, 2, 67-71, 1963
Wright, Precision of Color Differences Derived from a Multidimensional
Scaling Experiment, JOSA, 55, 12, 1650-1654, 1965
Mullen, The contrast sensitivity of human colour vision to red-green and
blue-yellow, J Physiol, 359, 381-400, 1985
Braje, Human efficiency for recognizing and detecting low-pass filtered
objects, Vision Res, 35, 21, 2955-2966, 1995
Fleury, Thesis, PhD, Dynamic Scheme Selection in Image Coding, Lausanne,
EPFL, 1999
Rubin, Using color and grayscale images to teach histology to
color-deficient medical students, Anatomical Sci Educ, 2, 2, 84-88, 2009
MacDonald & Luo: Colour Imaging, Vision and Technology.1999.
Brockmann, The unbearable distraction of color, IEEE Trans Prof Commun, 34,
3, 153-159, 1991

.........................................
Andrew Anthony "Tony" Havics, CHMM, CIH, PE
Environmental, Health & Safety, Microscopy, & Forensic Engineering
pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
(317) 718-7020 Office
(317) 718-7038 Fax
(317) 409-3238 Cell
aahavics-at-pH2LLC.com
www.ph2LLC.com

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Monday, April 08, 2013 10:38 AM
To: ph2-at-sprynet.com

Not a microscope but used in labs that might examine samples prepared by the
Ecomet with a microscope, so... Might anyone have circuit diagrams for an
ECOMET 3?

Thanks,

Ronald L'Herault

Lab Supervisor, Biomaterials Division
B.U. School of Dental Medicine
801 Albany Street S203
Roxbury, MA 02119

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But who decides what hue(s) are "natural" and "appropriate" for each
structure?

Different researchers might decide to use different colors to illustrate the
same structure, depending on their own aesthetics and preconception.
Differential coloring of the same structure in different publications has
the potential to distract, distort and confuse.

Also artificial coloring constitutes image manipulation. Once an
investigator imparts structural boundaries using artificial color, it
becomes more difficult if not impossible for others to visualize that image
from a different perspective. That is, the audience can be easily led by the
artificial rendering towards a particular line of interpretation or a
particular way of quantification. This is not desirable for the sake of
scientific objectiveness.

To meet rigorous scientific journal standards both the original grayscale
and the colored images need to be published side by side.

--
Connie C.W. Hsia, M.D.
University of Texas Southwestern Medical Center
Dallas, Texas
Connie.Hsia-at-utsouthwestern.edu

On 4/8/13 9:43 AM, "eikonika-at-otenet.gr" {eikonika-at-otenet.gr} wrote:

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} Hi there
}
} I think the (photoshop or similar) color on SEM micrographs once added helps
} later other people, including scientists, to distinguish structures that
} will take time to see without color. Nature loves camouflage; for instance
} sperm tails on the surface of a human egg (zona pellucida) are very similar
} to the structure of the zona and difficult to identify. Same is with
} cellular projections that spread over other cells etc. It could be an option
} for scientific journals to accept colored SEM micrographs if submitted along
} with the originals and checked for accuracy.
}
} Cheers
}
} yorgos
}
}
} ----- Original Message -----
} X-from: {dmitry.v.sokolov-at-gmail.com}
} To: {eikonika-at-otenet.gr}
} Sent: Monday, April 08, 2013 4:34 PM
} Subject: [Microscopy] Re: A new way to visualize specimen in
}
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver
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} }
} } Dear Stefan,
} }
} } thank you for the links. The images and movies are fascinating indeed.
} }
} } I understand how important the appealing images are for the success in
} } science:
} } http://confocal-manawatu.pbworks.com/w/page/63945076/How%20to%20Be%20Successf
} } ul%20in%20Science
} }
} } However, I am doubtful about the scientific significance of the
} } artificially coloured images unless the colour contrast represents
} } variations or differences in physical / chemical properties where the
} } fluorescence and Raman scattering microscopy or hyperspectral imaging
} } for material characterisation could be the examples.
} }
} } If we touch the technological base of the imaging as the source of the
} } primary data for scientific knowledge:
} } http://confocal-manawatu.pbworks.com/w/page/16347068/Technology%20of%20Resear
} } ch
} } colouring the images (unless it leads to the production of new
} } knowledge) is another technological operation. It adds time to the
} } technological sequence of image acquisition and/or analysis and
} } therefore reduces the efficiency of research.
} }
} } Unfortunately, I have a little experience in colouring of SEM images and
} } I would appreciate your comments very much.
} }
} } With kind regards,
} } Dmitry
} }
} } Advanced Knowledge Management
} } for MICROSCOPY and Image Analysis
} } ________________________________
} } Dmitry Sokolov, Ph.D.
} } Mob: +64 21 063 5382
} } dmitry.v.sokolov-at-gmail.com
} }
} } On 09.04.2013 0:50, stefan.diller-at-t-online.de wrote:
} } } ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } } America
} } } To Subscribe/Unsubscribe --
} } } http://www.microscopy.com/MicroscopyListserver
} } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } } ----------------------------------------------------------------------------
} } }
} } } Dear All,
} } } since some years I worked on a new concept of visualizing specimen in the
} } } scanning electron microscope.
} } }
} } } My profession as a photographer leads me first to search for
} } } scientifically correct but also aesthetically appealing images,
} } } which makes me believe, that the scientific community and with it every
} } } researcher should be committed to share the aesthetics of
} } } the microworld with as many people as possible. In scanning electron
} } } microscopes (SEMs), data is usually shown using grayscale
} } } images. Colours can be added easily with additional detector signals.
} } } This procedure is widely accepted for communicating results
} } } on up-to-date research to newspapers, magazines and TV broadcasting
} } } outside the scientific world.
} } }
} } } In the past mostly still shots of the specimen had been shown, or noisy
} } } TV rate screen captures. I thought it would be worth to
} } } bring the effects of a good movie into the SEM: fluent movement of the
} } } specimen, changing colour and lighting effects during the
} } } sequence, chances of focus and magnification.
} } }
} } } Find out more at my website www.nanoflight.info
} } } For those of you who like to see immediately, what`s this all about, go
} } } to
} } } www.nanoflight.info/nanoflight2.html
} } } directly and hope, that the server keeps running ;-)
} } }
} } } I am open to any discussion.
} } }
} } } All the best,
} } } Stefan
} } }
} } }
} }
} }
} } ==============================Original
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________________________________

UT Southwestern Medical Center
The future of medicine, today.

==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Hello
Clearly, the scientist has something in his mind when he takes microscopy
photos and chooses his field, i.e. one or two out of the myriad aspects
found in his specimen. The same is true for any researcher, whether he
analyzes data or perfom experiments and his results have the flavor of the
way he had looked at things. By colouring your SEM photos you can finalize
your results in a nice way and demonstrate what you had in your mind. In
this sense the word "image manipulation" sounds bit dogmatic. But I agree
that coloured and uncoloured photos could appear together. If your
scientific results are images, everybody can clearly see and judge the
"manipulation"...
Regards
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

----- Original Message -----
X-from: {Connie.Hsia-at-utsouthwestern.edu}
To: {eikonika-at-otenet.gr}
Sent: Monday, April 08, 2013 8:17 PM

If something is "difficult to identify" it SHOULD NOT BE COLORED! It can be misleading and manipulative.
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Monday, April 08, 2013 9:34 AM
To: Dusevich, Vladimir

Hi there

I think the (photoshop or similar) color on SEM micrographs once added helps later other people, including scientists, to distinguish structures that will take time to see without color. Nature loves camouflage; for instance sperm tails on the surface of a human egg (zona pellucida) are very similar to the structure of the zona and difficult to identify. Same is with cellular projections that spread over other cells etc. It could be an option for scientific journals to accept colored SEM micrographs if submitted along with the originals and checked for accuracy.

Cheers

yorgos

----- Original Message -----
X-from: {dmitry.v.sokolov-at-gmail.com}
To: {eikonika-at-otenet.gr}
Sent: Monday, April 08, 2013 4:34 PM

Hi Vladimir,
Although I usually appreciate your comments very much, I don't agree on that one.

Coloring a picture does not modify the content, it just make it easier to interpret.
The data in all papers are interpreted by the authors; this is not a big secret. Actually the authors even choose (wisely) the data to be presented! The objectivity of the choice made is in my opinion much more critical than the fact of coloring a picture. Especially for the field of microscopy where we usually can’t display the results as statistical numbers, choosing the right picture to represent a result is critical.
I think that it is good thing to present data in a way that make them easily understandable.

Regards,
Stephane

________________________________
X-from: "DusevichV-at-umkc.edu" {DusevichV-at-umkc.edu}
To: nizets2-at-yahoo.com
Sent: Monday, April 8, 2013 8:51 PM

If something is "difficult to identify" it SHOULD NOT BE COLORED! It can be misleading and manipulative.
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Monday, April 08, 2013 9:34 AM
To: Dusevich, Vladimir

Hi there

I think the (photoshop or similar) color on SEM micrographs once added helps later other people, including scientists, to distinguish structures that will take time to see without color. Nature loves camouflage; for instance sperm tails on the surface of a human egg (zona pellucida) are very similar to the structure of the zona and difficult to identify. Same is with cellular projections that spread over other cells etc. It could be an option for scientific journals to accept colored SEM micrographs if submitted along with the originals and checked for accuracy.

Cheers

yorgos

----- Original Message -----
X-from: {dmitry.v.sokolov-at-gmail.com}
To: {eikonika-at-otenet.gr}
Sent: Monday, April 08, 2013 4:34 PM

Dear All,
it had not been in my intend to start another discussion concerning the use of colour in SEM images.
What I had more in mind is presenting the MOVING images in high quality, with or without colours.

I use colours to enhance the aesthetical view of the image data but I gladly accept to show the videos in black and white in a
publication also, if it helps the work per se.

Have a look at this video please: http://vimeo.com/63638612
The right half is the exactly same video stream like in the left half, it only has no colour saturation...

At that point you might also argue that it should be said that the black and white image is a composit of one SE image channel and
three backscattered image channels and that this needs to be said in the headline accompaning the video. Sure, no problem.

In my opinion the video yields more information than a series of still shots of the same specimen. That`s why I am doing this. Not
because of the nice artificial colours, which I try to use in that way to give a more "natural" impression of the specimen.

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 09.04.13 10:11, schrieb nizets2-at-yahoo.com:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi Vladimir,
} Although I usually appreciate your comments very much, I don't agree on that one.
}
} Coloring a picture does not modify the content, it just make it easier to interpret.
} The data in all papers are interpreted by the authors; this is not a big secret. Actually the authors even choose (wisely) the data to be presented! The objectivity of the choice made is in my opinion much more critical than the fact of coloring a picture. Especially for the field of microscopy where we usually can�t display the results as statistical numbers, choosing the right picture to represent a result is critical.
} I think that it is good thing to present data in a way that make them easily understandable.
}
} Regards,
} Stephane
}
}
} ________________________________
} X-from: "DusevichV-at-umkc.edu" {DusevichV-at-umkc.edu}
} To: nizets2-at-yahoo.com
} Sent: Monday, April 8, 2013 8:51 PM
} Subject: [Microscopy] RE: A new way to visualize specimen in
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} If something is "difficult to identify" it SHOULD NOT BE COLORED! It can be misleading and manipulative.
} Vladimir
}
} Vladimir M. Dusevich, Ph.D.
} Electron Microscope Lab Director
} (816) 235-2072
} 371 School of Dentistry
} 650 E. 25th Street
} Kansas City, MO 64108-2784
}
} -----Original Message-----
} X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
} Sent: Monday, April 08, 2013 9:34 AM
} To: Dusevich, Vladimir
} Subject: [Microscopy] A new way to visualize specimen in
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi there
}
} I think the (photoshop or similar) color on SEM micrographs once added helps later other people, including scientists, to distinguish structures that will take time to see without color. Nature loves camouflage; for instance sperm tails on the surface of a human egg (zona pellucida) are very similar to the structure of the zona and difficult to identify. Same is with cellular projections that spread over other cells etc. It could be an option for scientific journals to accept colored SEM micrographs if submitted along with the originals and checked for accuracy.
}
} Cheers
}
} yorgos
}
}
} ----- Original Message -----
} X-from: {dmitry.v.sokolov-at-gmail.com}
} To: {eikonika-at-otenet.gr}
} Sent: Monday, April 08, 2013 4:34 PM
} Subject: [Microscopy] Re: A new way to visualize specimen in
}
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe --
} } http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Dear Stefan,
} }
} } thank you for the links. The images and movies are fascinating indeed.
} }
} } I understand how important the appealing images are for the success in
} } science:
} } http://confocal-manawatu.pbworks.com/w/page/63945076/How%20to%20Be%20Successful%20in%20Science
} }
} } However, I am doubtful about the scientific significance of the
} } artificially coloured images unless the colour contrast represents
} } variations or differences in physical / chemical properties where the
} } fluorescence and Raman scattering microscopy or hyperspectral imaging
} } for material characterisation could be the examples.
} }
} } If we touch the technological base of the imaging as the source of the
} } primary data for scientific knowledge:
} } http://confocal-manawatu.pbworks.com/w/page/16347068/Technology%20of%20Research
} } colouring the images (unless it leads to the production of new
} } knowledge) is another technological operation. It adds time to the
} } technological sequence of image acquisition and/or analysis and
} } therefore reduces the efficiency of research.
} }
} } Unfortunately, I have a little experience in colouring of SEM images and
} } I would appreciate your comments very much.
} }
} } With kind regards,
} } Dmitry
} }
} } Advanced Knowledge Management
} } for MICROSCOPY and Image Analysis
} } ________________________________
} } Dmitry Sokolov, Ph.D.
} } Mob: +64 21 063 5382
} } dmitry.v.sokolov-at-gmail.com
} }
} } On 09.04.2013 0:50, stefan.diller-at-t-online.de wrote:
} } } ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of
} } } America
} } } To Subscribe/Unsubscribe --
} } } http://www.microscopy.com/MicroscopyListserver
} } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } } ----------------------------------------------------------------------------
} } }
} } } Dear All,
} } } since some years I worked on a new concept of visualizing specimen in the
} } } scanning electron microscope.
} } }
} } } My profession as a photographer leads me first to search for
} } } scientifically correct but also aesthetically appealing images,
} } } which makes me believe, that the scientific community and with it every
} } } researcher should be committed to share the aesthetics of
} } } the microworld with as many people as possible. In scanning electron
} } } microscopes (SEMs), data is usually shown using grayscale
} } } images. Colours can be added easily with additional detector signals.
} } } This procedure is widely accepted for communicating results
} } } on up-to-date research to newspapers, magazines and TV broadcasting
} } } outside the scientific world.
} } }
} } } In the past mostly still shots of the specimen had been shown, or noisy
} } } TV rate screen captures. I thought it would be worth to
} } } bring the effects of a good movie into the SEM: fluent movement of the
} } } specimen, changing colour and lighting effects during the
} } } sequence, chances of focus and magnification.
} } }
} } } Find out more at my website www.nanoflight.info
} } } For those of you who like to see immediately, what`s this all about, go
} } } to
} } } www.nanoflight.info/nanoflight2.html
} } } directly and hope, that the server keeps running ;-)
} } }
} } } I am open to any discussion.
} } }
} } } All the best,
} } } Stefan
} } }
} } }
} }
} }
} } ==============================Original
} } Headers==============================
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16, 23 -- From stefan.diller-at-t-online.de Tue Apr 9 03:57:29 2013
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear All,

While Stefan Diller's images and movies are fascinating, they correspond to showing a grey object that is illuminated with colorized lamps. The 3D effect is nicely improved by the additional color channels, and these images and movies are indeed captivating.

This is different than artificially colorizing a SEM image, which ideally would be done in a manual way. In such a time-consuming process, the colorization can be used to add scientific knowledge and artistic skills into the image, and could give suitable cover illustrations of a scientific journal, while the (grey-value) raw data are shown in the article within the journal. Such colorized images are not to be seen as visualization of scientific results, but rather as communication of elsewhere documented scientific results to a lay audience. This is for example excellently done at http://www.micronaut.ch, as shown here:
http://www.micronaut.ch/category/show/new

All the best,

Henning.

Henning Stahlberg, PhD
Prof. for Structural Biology, C-CINA, Biozentrum, University Basel
Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland
http://c-cina.org | Tel. +41-61-387 32 62

On Apr 8, 2013, at 2:49 PM, {stefan.diller-at-t-online.de}
{stefan.diller-at-t-online.de} wrote:

}
}
}
} ----------------------------------------------------------------------------
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}
} Dear All,
} since some years I worked on a new concept of visualizing specimen in the scanning electron microscope.
}
} My profession as a photographer leads me first to search for scientifically correct but also aesthetically appealing images,
} which makes me believe, that the scientific community and with it every researcher should be committed to share the aesthetics of
} the microworld with as many people as possible. In scanning electron microscopes (SEMs), data is usually shown using grayscale
} images. Colours can be added easily with additional detector signals. This procedure is widely accepted for communicating results
} on up-to-date research to newspapers, magazines and TV broadcasting outside the scientific world.
}
} In the past mostly still shots of the specimen had been shown, or noisy TV rate screen captures. I thought it would be worth to
} bring the effects of a good movie into the SEM: fluent movement of the specimen, changing colour and lighting effects during the
} sequence, chances of focus and magnification.
}
} Find out more at my website www.nanoflight.info
} For those of you who like to see immediately, what`s this all about, go to
} www.nanoflight.info/nanoflight2.html
} directly and hope, that the server keeps running ;-)
}
} I am open to any discussion.
}
} All the best,
} Stefan
}
}
} --
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 Phone
} ++49-931-7848701 Fax
} ++49-175-7177051 Mobile
}
} Websites:
} www.nanoflight.info
} www.stefan-diller.com
} www.electronmicroscopy.info
} www.elektronenmikroskopie.info
} www.zwillingsprojekt.de
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Name: Ravi Thakkar

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Title-Subject: [Filtered] TEM grid analysis for TEM & SEM purpose.

Message: Dear Listeners,
I have a TEM grid, which is used for SEM analysis. Now I want to re-use this grid for TEM analysis.?
So, how to make TEM grid to usable for TEM analysis.?

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Hi Stephane,

You have written “The data in all papers are interpreted by the authors; this is not a big secret. Actually the authors even choose (wisely) the data to be presented!” Unfortunately for some of the papers “wisely” should be replaced on “wishfully”. Electron microscopy data are prone to abuse (in most cases unintentional abuse). In my field I tend to check presented data critically. Highlighting regions of low contrast with color can make this checking impossible and convince me (may be wrongfully) that data was manipulated. Even worse if some features are highlighted and others, very similar, left in gray levels. I have nothing against highlighting some obvious features, but why highlight something that stands apart anyway?

Regards,

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
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Sent: Tuesday, April 09, 2013 3:07 AM
To: Dusevich, Vladimir
Cc: microscopy-at-microscopy.com

If something is "difficult to identify" it SHOULD NOT BE COLORED! It can be misleading and manipulative.
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Monday, April 08, 2013 9:34 AM
To: Dusevich, Vladimir

Hi there

I think the (photoshop or similar) color on SEM micrographs once added helps later other people, including scientists, to distinguish structures that will take time to see without color. Nature loves camouflage; for instance sperm tails on the surface of a human egg (zona pellucida) are very similar to the structure of the zona and difficult to identify. Same is with cellular projections that spread over other cells etc. It could be an option for scientific journals to accept colored SEM micrographs if submitted along with the originals and checked for accuracy.

Cheers

yorgos

----- Original Message -----
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To: {eikonika-at-otenet.gr}
Sent: Monday, April 08, 2013 4:34 PM

Hello Collective!

Kind of a perplexing problem here: we are having difficulty with good infiltration of intestine and pancreas tissue samples. After one bad run, we greatly extended our infiltration steps (Epon/Spurr's resin) and had a repeat of bad results. We use microwave fixation and infiltration and it has been consistent and reliable with almost all specimens we have used it with.

In the current problematic batches, a couple samples infiltrated fine, while others in the same run did not. Interestingly enough, the control set of one tissue(intestine) infiltrated poorly, while the treated sets looked fine. In the pancreas, the treated tissue infiltrated poorly, while the control was good. Both sets were put through eight (8!) changes of pure resin, including of course some overnighters.

We're going to keep banging away at this, but does anyone have any sudden flashes of insight? You know, the "Hey that happened to me, too, and this is what we did!" sort?

Thanks------again.

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211

Phone: 573-884-5383
Email: tindallr-at-missouri.edu
Day job: www.emc.missouri.edu
Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
Blog: http://nadiasyard.com

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Hi Randy,
I had some problems with pancreas last year. Some areas of the tissue
block just crumbled. I use a standard Epon equivalent resin and standard
ethanol dehydration, propylene oxide infiltration protocol. Eventually I
decided that the major problem was related to fixation. Subsequently I
personally made up the 2% formaldehyde, 2% glutaraldehyde in 0.1M PO4
and attended the perfusion so I could mince the tissue into 1mm blocks
as soon as possible. I left the samples in fix overnight of longer. The
results were much better. I do not recall a problem with intestinal
tissue but that has been many years ago.
Good luck,
Larry

On 4/9/2013 1:19 PM, tindallr-at-missouri.edu wrote:
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} Hello Collective!
}
} Kind of a perplexing problem here: we are having difficulty with good infiltration of intestine and pancreas tissue samples. After one bad run, we greatly extended our infiltration steps (Epon/Spurr's resin) and had a repeat of bad results. We use microwave fixation and infiltration and it has been consistent and reliable with almost all specimens we have used it with.
}
} In the current problematic batches, a couple samples infiltrated fine, while others in the same run did not. Interestingly enough, the control set of one tissue(intestine) infiltrated poorly, while the treated sets looked fine. In the pancreas, the treated tissue infiltrated poorly, while the control was good. Both sets were put through eight (8!) changes of pure resin, including of course some overnighters.
}
} We're going to keep banging away at this, but does anyone have any sudden flashes of insight? You know, the "Hey that happened to me, too, and this is what we did!" sort?
}
} Thanks------again.
}
} Cheers,
} Randy
}
} Randy Tindall
} Senior EM Specialist
} Electron Microscopy Core Facility
} W122 Veterinary Medicine
} University of Missouri
} Columbia, MO 65211
}
} Phone: 573-884-5383
} Email: tindallr-at-missouri.edu
} Day job: www.emc.missouri.edu
} Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
} Blog: http://nadiasyard.com
}
}
}
}
}
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--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, Diabetes & Endocrinology Research Center Microscopy Core
UCSF, Dept. of Anatomy, Rm S657
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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Name: tugce karakulak

Organization: iowa state university

Title-Subject: [Filtered] EDS quantification problem..

Message: Dear Microscopy Server Listers;

We have an INCA X-sight EDS detector from Oxford Instruments and we have a trouble to quantify the
data.The detector collects the signals and software labels the elements while acquiring.However;
when you want to quantify, it gives exactly this error:

"No lines could be found for quantification of the following elements:Al,Cr,Co,Ni. This is
probably due to the accelerating voltage being too low,quantification of spectrum 1 aborted."
Error 1940[4]

Although I rebooted both detector and microscopy and change the accelerating voltage; the problem
wasn't solved.

Has anyone seen this error before?

Any help is appreciated.

Thank you very much.

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Hello...

Normally if my microscope is not communicating by rs-232 is when I get an error like this. I have a Zeiss system with my INCA.

David

M. David Frey
Senior Application Engineer
Rensselaer Polytechnic Institute
Low Center for Industrial Inn.
Center for Integrated Electronics
110 8th Street
CII 4161 (Office)
CII 6015 (Packages and Mail)
Troy, NY 12180
518-276-3323 (office)
518-698-2288 (mobile)
freym2-at-rpi.edu

On Apr 9, 2013, at 8:59 PM, microscopylistserver-noreply-at-microscopy.com wrote:

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} Name: tugce karakulak
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} Organization: iowa state university
}
} Title-Subject: [Filtered] EDS quantification problem..
}
} Message: Dear Microscopy Server Listers;
}
} We have an INCA X-sight EDS detector from Oxford Instruments and we have a trouble to quantify the
} data.The detector collects the signals and software labels the elements while acquiring.However;
} when you want to quantify, it gives exactly this error:
}
} "No lines could be found for quantification of the following elements:Al,Cr,Co,Ni. This is
} probably due to the accelerating voltage being too low,quantification of spectrum 1 aborted."
} Error 1940[4]
}
} Although I rebooted both detector and microscopy and change the accelerating voltage; the problem
} wasn't solved.
}
} Has anyone seen this error before?
}
} Any help is appreciated.
}
} Thank you very much.
}
} Login Host: 129.186.196.196
} Listserver Email Form V - 20120416
} ---------------------------------------------------------------------------
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M. David Frey
Senior Application Engineer
Rensselaer Polytechnic Institute
Low Center for Industrial Inn.
Center for Integrated Electronics
110 8th Street
CII 4161 (Office)
CII 6015 (Packages and Mail)
Troy, NY 12180
518-276-3323 (office)
518-698-2288 (mobile)
freym2-at-rpi.edu

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Typically, with an integrated system, Inca will read the beam kV from a
SEM. It uses this information for quantification. Check to see if Inca
is reading the proper beam kV. If it is not and the value it "thinks"
you are using is too low, you will get the error condition. Not sure if
it is possible to manually input the beam voltage used. I have never
tied that.

Woody
albe24.com

On 4/9/2013 9:02 PM, microscopylistserver-noreply-at-microscopy.com wrote:
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} Email: karakulaktugce-at-gmail.com
} Name: tugce karakulak
}
} Organization: iowa state university
}
} Title-Subject: [Filtered] EDS quantification problem..
}
} Message: Dear Microscopy Server Listers;
}
} We have an INCA X-sight EDS detector from Oxford Instruments and we have a trouble to quantify the
} data.The detector collects the signals and software labels the elements while acquiring.However;
} when you want to quantify, it gives exactly this error:
}
} "No lines could be found for quantification of the following elements:Al,Cr,Co,Ni. This is
} probably due to the accelerating voltage being too low,quantification of spectrum 1 aborted."
} Error 1940[4]
}
} Although I rebooted both detector and microscopy and change the accelerating voltage; the problem
} wasn't solved.
}
} Has anyone seen this error before?
}
} Any help is appreciated.
}
} Thank you very much.
}
} Login Host: 129.186.196.196
} Listserver Email Form V - 20120416
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Whenever I ran into a problem that I could not easily solve I just called
Oxford and got tech assistance. They are really great about getting back
to you quickly and helping you work through the problem. Don't hesitate
to do this as accessible tech help is one of Oxford's strengths.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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10, 32 -- "karakulaktugce-at-gmail.com" {karakulaktugce-at-gmail.com}
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Dear Ravi,
Regarding re-use of a TEM grid that had been in the SEM, I cannot see any reason why you would need to do anything to it to "re-use" it in the TEM.�
Unless you coated it (seems�unlikely) for SEM�imaging, why not�just�put it back in the TEM?
The only issue I can�foresee�is that it might be�somewhat�more contaminated if your SEM is not very clean, than before SEM analysis. But this is unlikely to be an issue unless you are looking for very low�contrast�materials, or doing elemental analysis.
�Maybe some more experienced EM experts can see some other problems?�
Pete

____________________________________________________________________________
Atomic Force Microscopy
Dr Peter Eaton and Dr Paul West
Available through all good bookshops, or direct from Oxford University Press at:
http://ukcatalogue.oup.com/product/9780199570454.do http://afmhelp.com

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Sounds to me like you haven't cleared your Periodic Table from your last analysis.
Gary

Gary M. Easton, Pres.
SCANNERS CORPORATION

on the road, sent from my NEXUS 4

microscopylistserver-noreply-at-microscopy.com wrote:

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Dear Researchers:

We have been getting several requests for EM work, including EM
immunogold labeling on exosomes. These are 40-100 nm vesicles secreted by
mammalian cells. The origin of exosomes is believed to be the
multivesicular bodies. Exosomes are usually collected from the culture
medium by high speed centrifugation, and then prepared using negative
staining method for EM viewing.

I have seen some images of immunogold labeled membrane proteins on
exosomes in published manuscripts. According to the method description in
these manuscripts, the labeling was done on grid following the deposition
of the exosomes. However, gold particles in these images were all at the
edge of exosomes where the vesicle outline could be clearly seen. I find
this puzzling.These exosomes were prepared as whole-mount so the entire
vesicles should have been covered by membrane. I would expect the gold
particles to be throughout the exosome's membrane (e.g., on top of
vesicles as well) instead of being only at the edge. Can anyone think of
a reason why this should not have been the case?

Thank you in advance for your input.

Hong

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11, 38 -- From hyi-at-emory.edu Wed Apr 10 06:17:45 2013
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Hi Hong!

Interesting question. I suppose that this is not due to out-of-focus particles.
To help answering your question it would be interesting to know how the vesicles were prepared, if they were fixed on the grid for example.
If they were not, you can imagine that the vesicle collapse during drying and that what you see is actually the "squashed" vesicles on the support.
A little bit like a color bubble which would leave a round mark after blowing up on a surface.

Regards,
Stephane

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Dear Researchers:

� � � � We have been getting several requests for EM work, including EM
immunogold labeling on exosomes.� These are 40-100 nm vesicles secreted by
mammalian cells.� The origin of exosomes is believed to be the
multivesicular bodies. Exosomes are usually collected from the culture
medium by high speed centrifugation, and then prepared using negative
staining method for EM viewing.

� � � � I have seen some images of immunogold labeled membrane proteins on
exosomes in published manuscripts.� According to the method description in
these manuscripts, the labeling was done on grid following the deposition
of the exosomes.� However, gold particles in these images were all at the
edge of exosomes where the vesicle outline could be clearly seen.� I find
this puzzling.These exosomes were prepared as whole-mount so the entire
vesicles should have been covered by membrane.� I would expect the gold
particles to be throughout the exosome's membrane (e.g., on top of
vesicles as well) instead of being only at the edge.� Can anyone think of
a reason why this should not have been the case?

Thank you in advance for your input.

Hong

________________________________

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Hello Ravi,

I routinely do TEM analysis of materials on locator grids (copper grid with amorphous carbon film) that have first been analyzed in one of our SEMs. The TEM grid is lightly affixed to an SEM substrate for EDS and imaging, and is then carefully removed from the substrate for analysis of the same material in the TEM. The samples are not coated for the SEM work, and transfer to the TEM presents no problems. Unless there is some other aspect to your question that's not mentioned below, you should be able to examine a grid in the TEM after it's been in the SEM.

Best regards,

Elaine

*********************************************************************
Elaine F.Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail: eschumacher-at-mccrone.com
Web Site: www.mccrone.com
*********************************************************************
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Title-Subject: [Filtered] TEM grid analysis for TEM & SEM purpose.

Message: Dear Listeners,
I have a TEM grid, which is used for SEM analysis. Now I want to re-use this grid for TEM analysis.?
So, how to make TEM grid to usable for TEM analysis.?

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Hi

one thing that occurs to me is that if the exosomes are 40-100nm then the gold label is going to be 5nm or less. I suspect the physical difficulties of imaging small gold on the upper surface of a negatively stained particle would render it pretty near invisible. This however may not be true at the edges of the exosome where not only might there be less material to obscure the gold but any layer of gold particles may be enhanced by effectively looking at in side profile.

I can't guarantee that this is so in this case without knowing the nature of the gold particle size, the actual layer thickness of the exosomes (presumably an upper and lower envelope when viewed from above and the negative stain density) but it does sound as if it could be just a simple physical effect.

Incidentally may I bid this group a fond farewell because I am taking early retirement at the end of this April. I can't remember how long I have participated but suspect it must be 15 years (- is that possible?). Over the years you have greatly enhanced my microscopical experience and knowledge. I may consider re-registering from home if I feel a sudden rush of nostalgia but haven't decided yet.

So long and thanks (... for some strange reason I want to say "for all the fish").

Malcolm

Malcolm Haswell
Imaging Suite
Faculty of Applied Sciences
University of Sunderland
SUNDERLAND
SR1 3SD
UK
email: malcolm.haswell-at-sunderland.ac.uk

________________________________________
X-from: nizets2-at-yahoo.com [nizets2-at-yahoo.com]
Sent: 10 April 2013 12:52
To: Malcolm Haswell

Hi Hong!

Interesting question. I suppose that this is not due to out-of-focus particles.
To help answering your question it would be interesting to know how the vesicles were prepared, if they were fixed on the grid for example.
If they were not, you can imagine that the vesicle collapse during drying and that what you see is actually the "squashed" vesicles on the support.
A little bit like a color bubble which would leave a round mark after blowing up on a surface.

Regards,
Stephane

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Sent: Wednesday, April 10, 2013 1:21 PM

Dear Researchers:

We have been getting several requests for EM work, including EM
immunogold labeling on exosomes. These are 40-100 nm vesicles secreted by
mammalian cells. The origin of exosomes is believed to be the
multivesicular bodies. Exosomes are usually collected from the culture
medium by high speed centrifugation, and then prepared using negative
staining method for EM viewing.

I have seen some images of immunogold labeled membrane proteins on
exosomes in published manuscripts. According to the method description in
these manuscripts, the labeling was done on grid following the deposition
of the exosomes. However, gold particles in these images were all at the
edge of exosomes where the vesicle outline could be clearly seen. I find
this puzzling.These exosomes were prepared as whole-mount so the entire
vesicles should have been covered by membrane. I would expect the gold
particles to be throughout the exosome's membrane (e.g., on top of
vesicles as well) instead of being only at the edge. Can anyone think of
a reason why this should not have been the case?

Thank you in advance for your input.

Hong

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33, 38 -- exosomes
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Dear Hong, dear all,
I would not trust these results too much, nor the interpretations - for a good reason:
I think that label-fracture and/or fracture-label is far more powerful for this type of specimen. Or, you directly go to cryo-TEM, with all Pro's and Con's.
kind regards,
Reinhard

--
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

next microscopy conferences:
http://www.mc2013.de/
MC2013 in Regensburg, Germany
http://www.imc2014.com/
18th IMC 2014 in Prague, Czech Rep.

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First check to see if the element list is fixed and has the correct elements in it or select current spectrum.

If that does not solve the problem enter configure and select INCA auto select the lines as this feature maybe turned off.

If this does not work call Oxford.

Keith Collins
USDOE
NETL-Albany
Albany, Oregon

Organization: iowa state university

Title-Subject: [Filtered] EDS quantification problem..

Message: Dear Microscopy Server Listers;

We have an INCA X-sight EDS detector from Oxford Instruments and we have a trouble to quantify the
data.The detector collects the signals and software labels the elements while acquiring.However;
when you want to quantify, it gives exactly this error:

"No lines could be found for quantification of the following elements:Al,Cr,Co,Ni. This is
probably due to the accelerating voltage being too low,quantification of spectrum 1 aborted."
Error 1940[4]

Although I rebooted both detector and microscopy and change the accelerating voltage; the problem
wasn't solved.

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4, 20 -- From: "Keith Collins" {Keith.Collins-at-netl.doe.gov}
4, 20 -- To: {karakulaktugce-at-gmail.com} , {Microscopy-at-microscopy.com}
4, 20 -- Subject: EDS quantification problem
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Howdy all,

Three people have now suggested that the uneven fields in our microwaves may be the culprit in our strange infiltration problems, so it's time to get serious about checking this out. We use Pelco Biowaves, which have been consistently reliable workhorses for years, with the Coldspot water recirculator. We'll do some testing with neon bulbs and check for bubbles in the water bath, etc.

Thanks to all who have replied so far!

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211

Phone: 573-884-5383
Email: tindallr-at-missouri.edu
Day job: www.emc.missouri.edu
Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
Blog: http://nadiasyard.com

==============================Original Headers==============================
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Hi everybody, I would like to bring to your attention slight
modifications to our

May 29 and 30 Meeting "Advances in Structural and Chemical Imaging" at
CAMCOR, U of Oregon, Eugene, OR, meeting

we hope many of you can attend since there is a zero participation
fee, but free coffee, bakery basket, lunch, finger food and drinks at
the poster session. We plan to have up to 25 posters in addition to
our 22 invited speakers.

For registration (and much more information), go to
http://nanocrystallography.net/ASCI2013

Posters need to be registered there separately. So far confirmed
vendor participation by: NanoMEGAS, Appfive, Eclipse Technologies, and
Tescan

------------------------------------------------------------------------------------------------------------

Advances in Structural and Chemical Imaging (ASCI 2013)

(A PREMIER* Network Conference), CAMCOR, Eugene, OR, May 29 - 30, 2013
(* Pooled Resources for Electron Microscopy Informatics Education and Research)

Day 1

Courtesy shuttle from Valley River Inn hotel to meeting venue 7:15 am,
bakery basket and coffee at 7:45 am (provided by PREMIER)

8.15-8.30 Welcome and Introduction to ASCI and the PREMIER Network �
Nigel D. Browning, PNNL

Session 1 Advances in Instrumentation I - Nigel D. Browning, PNNL (Chair)

8.30-9.00 Vortex Beams � Ben McMorran, U. Oregon
9.00-9.30 The UTEM project � Bernd Kabius, PNNL
9.30-10.00 Enabling observations of protein structural dynamics with
fast pump-probe electron microscopy and x-ray diffraction � James
Evans, PNNL

10.00-10.30 Coffee Break (provided by PREMIER)

10.30-11.00 Single molecule super-resolution microscopy for cancer
biology / Correlative Light and Electron Microscopy � Xiaolin Nan,
OHSU
11.00-11.30 Multiscale Imaging of Cells: From Organelle Systems to
Molecular Complexes � Gina Sosinsky, UC San Diego
11.30-12.00 Tomography of Catalysts � Ilke Arslan, PNNL

12.00-1.30 pm Lunch Break (provided by PREMIER)

Imaging Methods � Session 2 Dave C. Johnson, U. Oregon (Chair)

1.30-2.00 Contrast Improvement for Field Enhancement Based Near-field
Imaging � Erik S�nchez, PSU
2.00-2.30 The Debye-Waller factor of graphene � Chris Regan, UCLA
2.30-3.00 Imaging of Viruses by Cryogenic Transmission Electron
Microscopy and Three-Dimensional Reconstruction � David Belnap, U of
Utah

3.00-3.30 Coffee Break (provided by PREMIER)

3.30-4.00 Time-Resolved Electrostatic Force Microscopy of Thin Film
Solar Cells � David S. Ginger, UW
4.00-4.30 Imaging of Nanoparticle Dynamics on Oxide Surfaces � Abhaya Datye, UNM
4.30-5.00 Forisomes-Plant Derived ATP Independent Proteins Actuators �
Michael Knoblauch, WSU
5:00-5:30 Exploring Chemical and Structural Parameter Space - Thomas Vogt, USC

5.30-7.30 Poster Session

Day 2

Courtesy shuttle from Valley River Inn hotel to meeting venue 7:45 am,
bakery basket and coffee at 8:00 am (provided by PREMIER)

Session 1 Applications of Microscopy I � Erik S�nchez, PSU, (Chair)

8.30-9.00 Microstructure in nanomagnetism - open questions and
challenges � Kannan Krishnan, UW
9.00-9.30 Can you see me now? The power of analytical
aberration-corrected scanning transmission electron microscopy �
Robert F. Klie, UIC
9.30-10.00 Characterization of ODS alloys by SPS - A combinational TEM
and APT study � Yaqiao Wu, BSU

10.00-10.30 Coffee Break (provided by PREMIER)

10.30-11.00 Atom Probe - Brian Gorman, CSM
11.00-11.30 Bicrystallography for structural grain-boundary research �
Bryant York, PSU
11.30-12.00 Applied crystallography in SPM and TEM & open-access
crystallographic databases, Peter Moeck, PSU

12.00-1.30 pm Lunch Break (provided by PREMIER)

Session 2 Applications of Microscopy II � Peter Moeck, PSU (Chair)

1.30-2.00 A possible application of biomimetics in optimizing the
quantum efficiency of photovoltaics � Andreas Holzenburg, Texas A&M

2.00-2.30 Structural Analysis of Organic Perovskites and Bimetallic
Carbides for Photovoltaic Applications � Erwin M. Sabio, U. Wyoming

2.30-3.00 Diffraction Tomography & Automated Crystallite Orientation
and Phase Imaging in TEM, Amith Darbal, NanoMEGAS

3.00-3.15 Coffee Break (provided by PREMIER)

3.15-5.00 PREMIER Network Business Meeting

------------------------------------------------------------------

kind regards, Peter

--
Peter Moeck, PhD (Dr. rer. nat. Crystallography)
Associate Professor of Physics

Nano-Crystallography Group:
http://nanocrystallography.research.pdx.edu/nano-crystallography-group.
Free (as in free speech rather than free beer) crystallographic
information and interactive 3D visualizations of crystal structures
and morphologies with educational relevance:
http://nanocrystallography.research.pdx.edu/databases.

Portland State University
Department of Physics
P.O. Box 751, Portland, OR 97207-0751
Tel.: 503 725 4227, Departmental Fax.: 503 725 2815
pmoeck-at-pdx.edu
www.physics.pdx.edu/~pmoeck

Office 404, Science Research and Teaching Center
1719 SW 10th Ave, Portland, OR 97201

Office hours during quarters Tuesdays and Thursdays 12:00 to 12:30 and
per appointment

North American mirror of the Crystallography Open Database, COD,
http://nanocrystallography.org, with more than 220,000 carefully
evaluated datasets for download and interactive visualization in 3D,
for more interactive 3D display options try:
http://nanocrystallography.research.pdx.edu/search/codmirror/
Webportal: Open Access Crystallography Resource Portal
http://nanocrystallography.net.

"... no hell below us, above us only sky ..." John Lennon
"... der wackere Schwabe forcht sich nit ..." Albert Einstein
(inspired by Ludwig Uhland's poem)
"Was auch immer geschieht: Nie duerft ihr so tief sinken, von dem
Kakao, durch den man euch zieht, auch noch zu trinken." Erich Kaestner
"... the only thing needed for a happy life is a project one is
enthusiastic about." John C. H. Spence, Acta Cryst. A 69 (2013) 25-33

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Sorry if this is a duplicate message since I was having problems with
access to listserver.

Hong,

One possible explanation for the presence of label only around the
periphery of the spheres may be due to surface tension
or capillarity, or the tendency of liquids (and suspensions of gold)
to collect inside of folds or the interface between solids
(such as the contact of the sphere with the grid substrate). This is
the basis for negative staining, in fact.

Years ago, we did immunological staining on acrosomal vesicles and
localized enzymes on the surfaces of the vesicles.
The entire sphere (not just the edges) was labeled.

The only way to answer this for sure is to run a negative control with
spheres that lack the proteins in question. Hopefully,
the researchers did this.

--

John J. Bozzola, Ph.D., Professor
Gratefully Retired Director of IMAGE
Integrated Microscopy & Graphics Expertise
Southern Illinois University
Carbondale, IL 62901

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Hi all,
Does anyone know the proper knife angle for a sapphatome knife? 2 or 4? The box with info on it has gone missing.
thanks,
Beth

==============================Original Headers==============================
2, 21 -- From beth-at-plantbio.uga.edu Thu Apr 11 09:29:29 2013
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2, 21 -- From: Beth Richardson {beth-at-plantbio.uga.edu}
2, 21 -- Content-Type: text/plain; charset=us-ascii
2, 21 -- Subject: Sakura sapphatome knife
2, 21 -- Date: Thu, 11 Apr 2013 10:29:29 -0400
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I have inherited three of these knives in boxes and none of them state any
information other than the number of the knife.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E � Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-nhlbi.nih.go {mailto:connellyps-at-nhlbi.nih.gov} v

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely
shared
and reproduced.

On 4/11/13 10:40 AM, "beth-at-plantbio.uga.edu" {beth-at-plantbio.uga.edu} wrote:

}
} Hi all,
} Does anyone know the proper knife angle for a sapphatome knife? 2 or 4?
} The box with info on it has gone missing.
} thanks,
} Beth
}
}
} ==============================Original
} Headers==============================
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} 2, 21 -- Subject: Sakura sapphatome knife
} 2, 21 -- Date: Thu, 11 Apr 2013 10:29:29 -0400
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Hi Hong,
There is another possibility, but not a likely one. If the proteins
are mobile within the membrane of the liposome, they might migrate to
a region of higher curvature, so during centrifugation they could be
redistributed to the edges of flattened liposomes. If you know how
many labels there should be per liposome, you could calculate the
fraction you see, which will tell you whether you're seeing them all
or whether there are really unseen ones away from the edge. However,
since it may happen that label could be lost during preparation, this
might not be definitive.
Yours,
Bill

P.s., Malcolm, you will be missed.
BT

On Apr 10, 2013, at 5:56 AM, malcolm.haswell-at-sunderland.ac.uk wrote:

}
} one thing that occurs to me is that if the exosomes are 40-100nm
} then the gold label is going to be 5nm or less. I suspect the
} physical difficulties of imaging small gold on the upper surface of
} a negatively stained particle would render it pretty near invisible.
} This however may not be true at the edges of the exosome where not
} only might there be less material to obscure the gold but any layer
} of gold particles may be enhanced by effectively looking at in side
} profile.
}
} I can't guarantee that this is so in this case without knowing the
} nature of the gold particle size, the actual layer thickness of the
} exosomes (presumably an upper and lower envelope when viewed from
} above and the negative stain density) but it does sound as if it
} could be just a simple physical effect.
}
} Incidentally may I bid this group a fond farewell because I am
} taking early retirement at the end of this April. I can't remember
} how long I have participated but suspect it must be 15 years (- is
} that possible?). Over the years you have greatly enhanced my
} microscopical experience and knowledge. I may consider re-
} registering from home if I feel a sudden rush of nostalgia but
} haven't decided yet.
}
} So long and thanks (... for some strange reason I want to say "for
} all the fish").
}
} Malcolm
}
} Malcolm Haswell
} Imaging Suite
} Faculty of Applied Sciences
} University of Sunderland
} SUNDERLAND
} SR1 3SD
} UK
} email: malcolm.haswell-at-sunderland.ac.uk
}
}
} ________________________________________
} X-from: nizets2-at-yahoo.com [nizets2-at-yahoo.com]
} Sent: 10 April 2013 12:52
} To: Malcolm Haswell
} Subject: [Microscopy] Re: Immunogold labeling of membrane proteins
} on exosomes
}
}
} Hi Hong!
}
} Interesting question. I suppose that this is not due to out-of-focus
} particles.
} To help answering your question it would be interesting to know how
} the vesicles were prepared, if they were fixed on the grid for
} example.
} If they were not, you can imagine that the vesicle collapse during
} drying and that what you see is actually the "squashed" vesicles on
} the support.
} A little bit like a color bubble which would leave a round mark
} after blowing up on a surface.
}
} Regards,
} Stephane
}
} ----- Original Message -----
} X-from: "hyi-at-emory.edu" {hyi-at-emory.edu}
} To: nizets2-at-yahoo.com
} Cc:
} Sent: Wednesday, April 10, 2013 1:21 PM
} Subject: [Microscopy] Immunogold labeling of membrane proteins on
} exosomes
}
}
}
}
}
} Dear Researchers:
}
} We have been getting several requests for EM work, including EM
} immunogold labeling on exosomes. These are 40-100 nm vesicles
} secreted by
} mammalian cells. The origin of exosomes is believed to be the
} multivesicular bodies. Exosomes are usually collected from the culture
} medium by high speed centrifugation, and then prepared using negative
} staining method for EM viewing.
}
} I have seen some images of immunogold labeled membrane
} proteins on
} exosomes in published manuscripts. According to the method
} description in
} these manuscripts, the labeling was done on grid following the
} deposition
} of the exosomes. However, gold particles in these images were all
} at the
} edge of exosomes where the vesicle outline could be clearly seen. I
} find
} this puzzling.These exosomes were prepared as whole-mount so the
} entire
} vesicles should have been covered by membrane. I would expect the
} gold
} particles to be throughout the exosome's membrane (e.g., on top of
} vesicles as well) instead of being only at the edge. Can anyone
} think of
} a reason why this should not have been the case?
}
} Thank you in advance for your input.
}
} Hong
}
}
}
} ________________________________
}
} This e-mail message (including any attachments) is for the sole use of
} the intended recipient(s) and may contain confidential and privileged
} information. If the reader of this message is not the intended
} recipient, you are hereby notified that any dissemination,
} distribution
} or copying of this message (including any attachments) is strictly
} prohibited.
}
} If you have received this message in error, please contact
} the sender by reply e-mail message and destroy all copies of the
} original message (including attachments).
}

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Our monthly new episode of What's It Made Of? is out now on youtube today, direct from Northwestern University. This time we're looking at a chrome vanadium wrench.
We hope that it's a fun/educational way to describe to non-scientific folks the kinds of things we do with electron microscopes, and to get people interested in science-type things. Let the sharing begin!

http://youtu.be/ld4JaVB0NgM

ERiC Jay Miller
Microscopy & Imaging Specialist
Electron Probe Instrumentation Center

Northwestern University
Mail: 2036 Cook Hall
Office: 1152 Cook Hall
2220 Campus Drive
Evanston, IL 60208-3108

ph: (847) 467-0789
fax: (847) 467-657

http://www.nuance.northwestern.edu

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Dear Colleagues, could you please navigate me where I can buy Hitachi SEM
4500 FEG cathode?
Thank you in advance
Andrei
____________________________________
Andrei Kolmakov PhD,
Associate Professor
Department of Physics,
Southern Illinois University
1245 Lincoln Dr., Neckers 491
SIUC, Carbondale IL 62901 USA
Phone office (618)-453-5212
Phone 407 lab (618)-453-6106
Phone 140 lab (618)-453-6011
http://www.physics.siu.edu/people/kolmakov/

==============================Original Headers==============================
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Hey Beth!

I just happen to have a sapphatome knife in the orginal box. The
paper work says "The optimum clearance angle for most Sapphatome knifes
is 3-5 degrees. The best suited clearacne angle, however, shall be
determined by the user."

I could scan and send the instructions if you wish but they generally
seem to follow diamond knife rules. The instructions want speeds slower
than 0.5mm/second, and thiner than 100nm (1000 Angstroms actually). It
says there is some "wetting Solution" if needed but there is no "wetting
solution" in in the box or listed on the packing list. Only two bottles of
cleaning solution.

It does suggest that if dirty clean it with "dense sulfuric acid" seems a
little extreme to me.

On 11 Apr 2013 at 9:39, beth-at-plantbio.uga.edu wrote:

}
}
}
} ----------------------------------------------------------------------
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} Hi all,
} Does anyone know the proper knife angle for a sapphatome knife? 2 or
} 4? The box with info on it has gone missing. thanks, Beth
}
}
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Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-miamioh.edu
http://www.cami.muohio.edu

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Hi Richard,
Thank you for the clearance angle info on the sapphatome knife (3-5 degrees). I greatly appreciated the responses I received from the listserv gang.
thanks again,
Beth

On Apr 16, 2013, at 4:58 PM, Richard E. Edelmann wrote:

} Hey Beth!
}
} I just happen to have a sapphatome knife in the orginal box. The
} paper work says "The optimum clearance angle for most Sapphatome knifes
} is 3-5 degrees. The best suited clearacne angle, however, shall be
} determined by the user."
}
} I could scan and send the instructions if you wish but they generally
} seem to follow diamond knife rules. The instructions want speeds slower
} than 0.5mm/second, and thiner than 100nm (1000 Angstroms actually). It
} says there is some "wetting Solution" if needed but there is no "wetting
} solution" in in the box or listed on the packing list. Only two bottles of
} cleaning solution.
}
} It does suggest that if dirty clean it with "dense sulfuric acid" seems a
} little extreme to me.
}
}
} On 11 Apr 2013 at 9:39, beth-at-plantbio.uga.edu wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------
} } ------ The Microscopy ListServer -- CoSponsor: The Microscopy Society
} } of America To Subscribe/Unsubscribe --
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} } ----------------------------------------------------------------------
} } ------
} }
} } Hi all,
} } Does anyone know the proper knife angle for a sapphatome knife? 2 or
} } 4? The box with info on it has gone missing. thanks, Beth
} }
} }
} } ==============================Original
} } Headers============================== 2, 21 -- From
} } beth-at-plantbio.uga.edu Thu Apr 11 09:29:29 2013 2, 21 -- Received: from
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} } -- Thu, 11 Apr 2013 10:29:27 -0400 2, 21 -- From: Beth Richardson
} } {beth-at-plantbio.uga.edu} 2, 21 -- Content-Type: text/plain;
} } charset=us-ascii 2, 21 -- Subject: Sakura sapphatome knife 2, 21 --
} } Date: Thu, 11 Apr 2013 10:29:29 -0400 2, 21 -- Message-Id:
} } {53E70F92-FA5C-4760-AD0F-B3E26214D771-at-plantbio.uga.edu} 2, 21 -- To:
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}
}
} Richard E. Edelmann, Ph.D., Director
} Center for Advanced Microscopy & Imaging
} 9C Upham Hall
} Miami University, Oxford, OH 45056
} Ph: 513.529.5712 Fax: 513.529.4243
} E-mail: edelmare-at-miamioh.edu
} http://www.cami.muohio.edu
}

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Organization: New England Society for Microscopy

Title-Subject: [Filtered] 30th Annual NESM Spring Symposium & Workshops at the MBL

Message: You are invited to the New England Society for Microscopy's 30th annual Spring Symposium &
Workshops, hosted by the Marine Biological Laboratory in Woods Hole, MA, on May 2nd-3rd. The meeting
will consist of Thursday afternoon workshops and Friday seminars, as well as a vendor session and
student poster competition on Friday. Registration closes on April 22nd at 12:00 PM. More
information and registration instructions are available on our website (www.nesmicroscopy.org). Keep
up with NESM on the web by following us on Facebook (http://www.facebook.com/NESMicroscopy) and on
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We hope to see you in May!

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Hi

I want to pass on some information I gathered at a recent California Community College Association for Occupational Education (CCCAOE) meeting.

At that meeting I met Greg Thomas, HR Director for the Foundation for California Community Colleges (FCCC). One of the services the FCCC offers is to handle all of the paper work and forms needed to establish student intern positions. They do it all, or just the part you need. Their services can range from advice to actually handling all the bookkeeping, insurance, credits, and payroll. Greg said they can make it as simple as the employer just writing a check to cover the costs.

Here at Delta we are always on the look out for training opportunities in the real world for our students. Sometimes the bureaucracy just gets in the way. With help from the FCCC establishing an internship should be much easier than ever. Please consider establishing intern opportunities for our students.

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Title-Subject: [Filtered] sem of beeswax

Message: Hi Is it possible to study beeswax under the scanning electron microscope and how they are
being studied under high vacuum

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2013 Yale Microscopy Workshop

Time: June 4 - 6th, 2013
Location: Yale Medical School, The Anlyan Center (TAC), New Haven, CT
Cost: Free registration (on-line or on-site)
Information: http://microscopy.med.yale.edu/index.aspx

Features:

- Symposium on Correlative Light and Electron Microscopy (CLEM)
- Open access to state-of-the-art confocal, super resolution and electron
microscopes, as well as various EM sample preparation equipment
- Demonstrations, technical lectures,and walk-in clinics
- Multiple vendor participation (} 10)

We invite you to participate in a three-day microscopy workshop. This free
annual event is a combination of symposium and
open access to a wide variety of fully functional microscopes. This year's
symposium is focused on Correlative Light and Electron Microscopy. It is
open to domestic and international research community and only requires
free registration.

This event is designed for the busy researcher who wants to explore new
techniques, see the latest in instrumentation, or simply get needed advice
from vendor representatives. Reservations can be made for many of the
instruments after registration. View this as an opportunity to perform a
"surgical strike" exploration without any financial or time commitments.
Feel free to come for advice from vendor representatives or to try out a
microscope even if you are not attending a class or a seminar.

Please see the website for free registration, a schedule of events, and
list of equipment. http://microscopy.med.yale.edu/index.aspx

Please feel free to disseminate information about this workshop at your
institution. Need to try out an instrument that is not on the list but
manufactured by a vendor that is already participating? Write to an
organizer and we'll see if they can bring it.

We look forward to seeing you there - please reserve early.

The organizers:

Ann Haberman ann.haberman-at-yale.edu
Derek Toomre derek.toomre-at-yale.edu
Joerg Bewersdorf joerg.bewersdorf-at-yale.edu
Xinran Liu xinran.liu-at-yale.edu

-------------------------------------------------

Ann Haberman, Ph.D.
Department of Laboratory Medicine
Yale University School of Medicine
(203) 785-7349 (office)

Derek Toomre, Ph.D.
Department of Cell Biology
Yale University School of Medicine
(203) 785-4319 (office)

Joerg Bewersdorf, Dr. rer. nat.
Department of Cell Biology
Yale University School of Medicine
(203) 737-5704 (office)

Xinran Liu, M.D. & Ph.D.
Department of Cell Biology, EM Core Facility
Yale University School of Medicine
(203) 785-4050 (office)

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The University of Pennsylvania Nanoscale Characterization Facility has two Gatan Dual Ion Mill 600s in need of a new home. One is almost functional and the other is good for parts. If you're interested, please contact me at tjford-at-seas.upenn.edu or 215-573-6123.

Jamie Ford
Staff Scientist
UPenn Nanoscale Characterization Facility

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Email: zluo-at-uncfsu.edu
Name: Zhiping Luo

Organization: Fayetteville State University

Title-Subject: [Filtered] Powder Pellet Press for microprobe

Message: Greetings!

We need to purchase a Pellet Press to prepare powder samples for the microprobe analysis. I very
appreciate for any recommendations on suitable vendors.

Sincerely,

Zhiping Luo
Fayetteville State University

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Email: henrik.kaker-at-guest.arnes.si
Name: Henrik Kaker

Organization: SEM-EDS Lab at Metal Ravne, Slovenia

Title-Subject: [Filtered] Jeol diffusion pump DP-4E

Message: Hello all,

We are trying to find the Jeol diffusion pump DP-4E for our SEM JSM35C. Has anyone had a any
information about this pump. Thank you for you help.

Best regards,

Henrik

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The Department of Chemical Engineering & Materials Science at UC Davis is
offering for sale a JEOL JEM 2500SE Transmission Electron Microscope.

Standard configuration as provided by JEOL, Inc.
200 kV Schottky emitter
Instrument continuously supported by JEOL service contract
Gatan Tridiem Imaging Filter
Gatan GIF UltraScan Camera US1000FT 1
Gatan Digiscan II Model 788
3 STEM detectors
Annular Dark Field and Bright field STEM detectors
Fischione Model 3000 Annular Dark Field Detector
Gatan First Light Camera Controller
Gatan MultiScan Retractable Camera (MSC Model 794)
Gatan Digitial Micrograph 2.0 Software, incl. computer and software
ThermoNoran EDX Detector System 6, Model #6719A, incl. computer and software
Chiller included

Please contact:
Prof. Klaus van Benthem
benthem-at-ucdavis.edu
with any questions.

Fred Hayes
Manager
Advanced Materials Characterization and Testing Lab (AMCaT)
Dept of Chemical Engineering and Material Science
3001 Ghausi Hall
Univ of CA Davis
Davis, CA 95616
530-752-0284
http://chms.engineering.ucdavis.edu/index.html
http://chms.engineering.ucdavis.edu/research/amcat/index.html

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I am interested in how people have replaced Polaroid Film (4X5) Image capture on their electron microscopes. (in order to avoid congestion on this server please respond to me directly at wa5ekh-at-juno.com)

Thanks!

Jeff Day
N. Texas (Dallas Area)
____________________________________________________________
Refinance Now 2.53% FIXED
No cost and no fee mortgage! No SSN rqd. Free quote in minutes. (2.53% APR)
http://thirdpartyoffers.juno.com/TGL3131/517184d538fb64d55f20st02vuc

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Dear Fellow listers,

For those of you who do fluorescence, Dr Ploem, inventor of the early
epi-fluorescence system known as the Ploemopak, is in the running for
a Nobel Prize. A special website has been set up on his invention.
Below is a note he recently sent to the New York Microscopical
Society.

Enjoy!
Barbara Foster
Microscopy/Microscopy Education
Bfoster-at-mme1.com
(972)924-5310
Join our current User Study: New Directions in Live Cell & Tissue Imaging
April 17-26. Visit MicroscopyEducation.com for details

Dear Mel Pollinger,
I have been mentioned on the Dutch media as a possible
candidate for a Nobel Price. Some members of the N.Y.M.S may be
interested in some details on the development of multi-wavelenghs
epifluorescence microscopy.

A comprehensive WEBSITE has been created for the National Dutch
Science Museum to illustrate the reason for the incorporation of my
early prototype fluorescence epiilluminators in the permanent
collection of that museum, as well as the exposition of the very first
Leitz-Leica epi-illumination fluorescence microscope with a Ploemopak
fluorescence epi-illuminator in the VISEUM science Museum in Wetzlar.
Germany.

Website:
www.ploem-fluorescence-microscopy.com
With kind regards
Johan Ploem, (J.S. Ploem)

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7, 27 -- Message-ID: {CAASiXrqv1TRbSR9heCb1htwzEtnu2h=xmqvEqdRCPrR7zXOLbw-at-mail.gmail.com}
7, 27 -- Subject: The Ploemopak to earn a Nobel Prize? ... and a special website
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Hi everybody, I would like to bring to your attention slight
modifications to our

May 29 and 30 Meeting "Advances in Structural and Chemical Imaging" at
CAMCOR, U of Oregon, Eugene, OR, meeting

we hope many of you can attend since there is a zero participation
fee, but free coffee, bakery basket, lunch, finger food and drinks at
the poster session. We plan to have up to 25 posters in addition to
our 22 invited speakers.

For registration (and much more information), go to
http://nanocrystallography.net/ASCI2013
Posters need to be registered there separately. So far confirmed
vendor participation by: NanoMEGAS, Appfive, Eclipse Technologies, and
Tescan

For free registration (and much more information), go to
http://nanocrystallography.net/ASCI2013

Posters need to be registered there separately. So far confirmed vendor
participation by NanoMEGAS, Appfive, and Eclipse Technologies,
sponsorship by FEI, JEOL, Tescan, NanoMEGAS, and ONAMI

------------------------------------------------------------------------------------------------------------
Advances in Structural and Chemical Imaging (ASCI 2013)

(A PREMIER* Network Conference), CAMCOR, Eugene, OR, May 29 - 30, 2013
(* Pooled Resources for Electron Microscopy Informatics Education and
Research)

Day 1

Courtesy shuttle from Valley River Inn hotel to meeting venue 7:15 am,
bakery basket and coffee at 7:45 am (provided by PREMIER)

8.15-8.30 Welcome and Introduction to ASCI and the PREMIER Network �
Nigel D. Browning, PNNL

Session 1 Advances in Instrumentation I - Nigel D. Browning, PNNL (Chair)

8.30-9.00 Vortex Beams � Ben McMorran, U. Oregon
9.00-9.30 The UTEM project � Bernd Kabius, PNNL
9.30-10.00 Enabling observations of protein structural dynamics with
fast pump-probe electron microscopy and x-ray diffraction � James
Evans, PNNL

10.00-10.30 Coffee Break (provided by PREMIER)

10.30-11.00 Single molecule super-resolution microscopy for cancer
biology / Correlative Light and Electron Microscopy � Xiaolin Nan,
OHSU
11.00-11.30 Multiscale Imaging of Cells: From Organelle Systems to
Molecular Complexes � Gina Sosinsky, UC San Diego
11.30-12.00 Tomography of Catalysts � Ilke Arslan, PNNL

12.00-1.30 pm Lunch Break (provided by PREMIER)

Imaging Methods � Session 2 Dave C. Johnson, U. Oregon (Chair)

1.30-2.00 Contrast Improvement for Field Enhancement Based Near-field
Imaging � Erik S�nchez, PSU
2.00-2.30 The Debye-Waller factor of graphene � Chris Regan, UCLA
2.30-3.00 Imaging of Viruses by Cryogenic Transmission Electron
Microscopy and Three-Dimensional Reconstruction � David Belnap, U of
Utah

3.00-3.30 Coffee Break (provided by PREMIER)

3.30-4.00 Time-Resolved Electrostatic Force Microscopy of Thin Film
Solar Cells � David S. Ginger, UW
4.00-4.30 Imaging of Nanoparticle Dynamics on Oxide Surfaces � Abhaya
Datye, UNM
4.30-5.00 Forisomes-Plant Derived ATP Independent Proteins Actuators �
Michael Knoblauch, WSU
5:00-5:30 Exploring Chemical and Structural Parameter Space - Thomas
Vogt, USC

5.30-7.30 Poster Session

Day 2

Courtesy shuttle from Valley River Inn hotel to meeting venue 7:45 am,
bakery basket and coffee at 8:00 am (provided by PREMIER)

Session 1 Applications of Microscopy I � Erik S�nchez, PSU, (Chair)

8.30-9.00 Microstructure in nanomagnetism - open questions and
challenges � Kannan Krishnan, UW
9.00-9.30 Can you see me now? The power of analytical
aberration-corrected scanning transmission electron microscopy �
Robert F. Klie, UIC
9.30-10.00 Characterization of ODS alloys by SPS - A combinational TEM
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------------------------

kind regards, Peter

--
Peter Moeck, PhD (Dr. rer. nat. Crystallography)
Associate Professor of Physics

Nano-Crystallography Group:
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Free (as in free speech rather than free beer) crystallographic information
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Portland State University
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Dear All,
anybody out to share a protocol for Osmiumtetroxide double fixing?

I need it to hopefully get rid of the specimen charging at large plant specimen for SEM which did not work up to now either with
"heavy" sputtercoating nor with an additional osmiumteroxide vapor fixation step.

Best regards,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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Title-Subject: [Filtered] How to process Bacteriophage for SEM ?

Message: Dear Listener,
How to process bacteriophage for SEM.?
Is there osmium treatment will help in SEM imaging.?

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Hi Stephan,
I've had good luck using the OTO method. Your times for your large plant
tissue may differ and you may need to adjust accordingly. Take note that the
tannic acid and thiocarbohydrazide need to made up fresh. They may be
buffered or not (I use aqueous).
-Fix in primary fixative (some add Tannic to their Glut, I do not.
-Rinse several times with buffer
-Fix in 1% osmium for 1 hour
-Immerse in either 1% tannic acid (TA) or saturated thiocarbohydrazide (TCH)
for at least 10 min-probably longer for large plant material
-Rinse THOROUGHLY with ddH2O
-Incubate in 1% Osmium tetroxide for 1 hour
-Rinse thoroughly with ddH2O
-You may stop here or repeat the last two steps, making sure to rinse
THOROUGHLY.
The TCH and TA are ligands which help the impregnation of osmium into
tissues.

Good Luck,
Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

-----Original Message-----
X-from: stefan.diller-at-t-online.de [mailto:stefan.diller-at-t-online.de]
Sent: Monday, April 22, 2013 1:30 AM
To: pekysar-at-ucdavis.edu

Dear All,
anybody out to share a protocol for Osmiumtetroxide double fixing?

I need it to hopefully get rid of the specimen charging at large plant
specimen for SEM which did not work up to now either with
"heavy" sputtercoating nor with an additional osmiumteroxide vapor fixation
step.

Best regards,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

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18, 30 -- From pekysar-at-ucdavis.edu Mon Apr 22 12:04:03 2013
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Stefan,

Those are good suggestions about fixation from Pat. You also might consider the size and shape of your samples. If the bottom surface that you mount to the stub is convex, so only the center is contacting the stub, you might not be making adequate electrical contact between the upper coated surface and the grounded stub. Sputter coating can have a difficult time getting into the gap between the edge of the sample and the stub, and the coated surface might not be grounded. One solution for this issue is to fill that gap with something like colloidal silver paint or colloidal graphite (DAG). If you coat after filling the gap, you can use something that is not conductive.

Good luck,

--John

John Chandler
EM Lab
Colorado School of Mines
jpchandl-at-mines.edu

On Apr 22, 2013, at 11:09 AM, {pekysar-at-ucdavis.edu}
wrote:

}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} Hi Stephan,
} I've had good luck using the OTO method. Your times for your large plant
} tissue may differ and you may need to adjust accordingly. Take note that the
} tannic acid and thiocarbohydrazide need to made up fresh. They may be
} buffered or not (I use aqueous).
} -Fix in primary fixative (some add Tannic to their Glut, I do not.
} -Rinse several times with buffer
} -Fix in 1% osmium for 1 hour
} -Immerse in either 1% tannic acid (TA) or saturated thiocarbohydrazide (TCH)
} for at least 10 min-probably longer for large plant material
} -Rinse THOROUGHLY with ddH2O
} -Incubate in 1% Osmium tetroxide for 1 hour
} -Rinse thoroughly with ddH2O
} -You may stop here or repeat the last two steps, making sure to rinse
} THOROUGHLY.
} The TCH and TA are ligands which help the impregnation of osmium into
} tissues.
}
} Good Luck,
} Pat Kysar, C.E.M.T.
} Chair, Certification Board, Microscopy Society of America
} University of California, Davis
} School of Medicine
} Dept of Pathology and Laboratory Medicine
} Electron Microscopy Lab
}
}
} -----Original Message-----
} X-from: stefan.diller-at-t-online.de [mailto:stefan.diller-at-t-online.de]
} Sent: Monday, April 22, 2013 1:30 AM
} To: pekysar-at-ucdavis.edu
} Subject: [Microscopy] Osmiumtetroxide - Thiocarbohydrazide - Osmiumteroxide
} Protocol
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Dear All,
} anybody out to share a protocol for Osmiumtetroxide double fixing?
}
} I need it to hopefully get rid of the specimen charging at large plant
} specimen for SEM which did not work up to now either with
} "heavy" sputtercoating nor with an additional osmiumteroxide vapor fixation
} step.
}
} Best regards,
} Stefan
}
}
} --
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 Phone
} ++49-931-7848701 Fax
} ++49-175-7177051 Mobile
}
} Websites:
} www.nanoflight.info
} www.stefan-diller.com
} www.electronmicroscopy.info
} www.elektronenmikroskopie.info
} www.zwillingsprojekt.de
} www.assisi.de
} Anfahrt: http://Mail.map24.com/Stefan.Diller
} -----------------------------------------------------
}
} ==============================Original Headers==============================
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} 18, 30 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com}
} 18, 30 -- Cc: {stefan.diller-at-t-online.de}
} 18, 30 -- Subject: RE: [Microscopy] Osmium tetroxide - Thiocarbohydrazide - Osmium teroxide Protocol
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11, 30 -- From jpchandl-at-mines.edu Mon Apr 22 12:28:01 2013
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11, 30 -- 22 Apr 2013 11:28:01 -0600
11, 30 -- From: John Chandler {jpchandl-at-mines.edu}
11, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} ,
11, 30 -- Pat Kysar
11, 30 -- {pekysar-at-ucdavis.edu}
11, 30 -- Subject: Re: [Microscopy] RE: Osmium tetroxide - Thiocarbohydrazide - Osmium
11, 30 -- teroxide Protocol
11, 30 -- Thread-Topic: [Microscopy] RE: Osmium tetroxide - Thiocarbohydrazide -
11, 30 -- Osmium teroxide Protocol
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Contents Retrieved from Microscopy Listserver Archives
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Dear All,
thanks a lot for this whole collection of protocols. If time permits I will try some different ones and come back in some weeks
with the results.

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 22.04.13 19:31, schrieb jpchandl-at-mines.edu:
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} Stefan,
}
} Those are good suggestions about fixation from Pat. You also might consider the size and shape of your samples. If the bottom surface that you mount to the stub is convex, so only the center is contacting the stub, you might not be making adequate electrical contact between the upper coated surface and the grounded stub. Sputter coating can have a difficult time getting into the gap between the edge of the sample and the stub, and the coated surface might not be grounded. One solution for this issue is to fill that gap with something like colloidal silver paint or colloidal graphite (DAG). If you coat after filling the gap, you can use something that is not conductive.
}
} Good luck,
}
} --John
}
} John Chandler
} EM Lab
} Colorado School of Mines
} jpchandl-at-mines.edu
}
}
}
} On Apr 22, 2013, at 11:09 AM, {pekysar-at-ucdavis.edu}
} wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} }
} } Hi Stephan,
} } I've had good luck using the OTO method. Your times for your large plant
} } tissue may differ and you may need to adjust accordingly. Take note that the
} } tannic acid and thiocarbohydrazide need to made up fresh. They may be
} } buffered or not (I use aqueous).
} } -Fix in primary fixative (some add Tannic to their Glut, I do not.
} } -Rinse several times with buffer
} } -Fix in 1% osmium for 1 hour
} } -Immerse in either 1% tannic acid (TA) or saturated thiocarbohydrazide (TCH)
} } for at least 10 min-probably longer for large plant material
} } -Rinse THOROUGHLY with ddH2O
} } -Incubate in 1% Osmium tetroxide for 1 hour
} } -Rinse thoroughly with ddH2O
} } -You may stop here or repeat the last two steps, making sure to rinse
} } THOROUGHLY.
} } The TCH and TA are ligands which help the impregnation of osmium into
} } tissues.
} }
} } Good Luck,
} } Pat Kysar, C.E.M.T.
} } Chair, Certification Board, Microscopy Society of America
} } University of California, Davis
} } School of Medicine
} } Dept of Pathology and Laboratory Medicine
} } Electron Microscopy Lab
} }
} }
} } -----Original Message-----
} } X-from: stefan.diller-at-t-online.de [mailto:stefan.diller-at-t-online.de]
} } Sent: Monday, April 22, 2013 1:30 AM
} } To: pekysar-at-ucdavis.edu
} } Subject: [Microscopy] Osmiumtetroxide - Thiocarbohydrazide - Osmiumteroxide
} } Protocol
} }
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } ----------------------------------------------------------------------------
} }
} } Dear All,
} } anybody out to share a protocol for Osmiumtetroxide double fixing?
} }
} } I need it to hopefully get rid of the specimen charging at large plant
} } specimen for SEM which did not work up to now either with
} } "heavy" sputtercoating nor with an additional osmiumteroxide vapor fixation
} } step.
} }
} } Best regards,
} } Stefan
} }
} }
} } --
} }
} }
} } -----------------------------------------------------
} } Stefan Diller - Scientific Photography
} } Arndtstrasse 22
} } D - 97072 Wuerzburg Germany
} } ++49-931-7848700 Phone
} } ++49-931-7848701 Fax
} } ++49-175-7177051 Mobile
} }
} } Websites:
} } www.nanoflight.info
} } www.stefan-diller.com
} } www.electronmicroscopy.info
} } www.elektronenmikroskopie.info
} } www.zwillingsprojekt.de
} } www.assisi.de
} } Anfahrt: http://Mail.map24.com/Stefan.Diller
} } -----------------------------------------------------
} }
} } ==============================Original Headers==============================
} } 8, 20 -- From stefan.diller-at-t-online.de Mon Apr 22 03:19:40 2013
} } 8, 20 -- Received: from mailout09.t-online.de (mailout09.t-online.de
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} } 18, 30 -- From: "Patricia Kysar" {pekysar-at-ucdavis.edu}
} } 18, 30 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com}
} } 18, 30 -- Cc: {stefan.diller-at-t-online.de}
} } 18, 30 -- Subject: RE: [Microscopy] Osmium tetroxide - Thiocarbohydrazide - Osmium teroxide Protocol
} } 18, 30 -- Date: Mon, 22 Apr 2013 10:03:30 -0700
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} 11, 30 -- From jpchandl-at-mines.edu Mon Apr 22 12:28:01 2013
} 11, 30 -- Received: from EXCH-CAS4.adit.mines.edu (exch-cas4.Mines.EDU [138.67.132.144])
} 11, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id r3MHS18U020783
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} 11, 30 -- 22 Apr 2013 11:28:01 -0600
} 11, 30 -- From: John Chandler {jpchandl-at-mines.edu}
} 11, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} ,
} 11, 30 -- Pat Kysar
} 11, 30 -- {pekysar-at-ucdavis.edu}
} 11, 30 -- Subject: Re: [Microscopy] RE: Osmium tetroxide - Thiocarbohydrazide - Osmium
} 11, 30 -- teroxide Protocol
} 11, 30 -- Thread-Topic: [Microscopy] RE: Osmium tetroxide - Thiocarbohydrazide -
} 11, 30 -- Osmium teroxide Protocol
} 11, 30 -- Thread-Index: AQHOP3wvjHZ2jGEl7EyU5C+YP2LliZji4tAA
} 11, 30 -- Date: Mon, 22 Apr 2013 17:28:00 +0000
} 11, 30 -- Message-ID: {E3AB3129-7237-40EB-856B-A4B6C8E64A0F-at-mines.edu}
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8, 23 -- From stefan.diller-at-t-online.de Mon Apr 22 12:36:39 2013
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8, 23 -- Date: Mon, 22 Apr 2013 19:36:34 +0200
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8, 23 -- - THANKS
8, 23 -- References: {201304221731.r3MHVBab027357-at-ns.microscopy.com}
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Contents Retrieved from Microscopy Listserver Archives
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Stefan, John has a valid point. It may just be your mounting. Another trick
I use to mount large samples on to the stub: I add a little graphite power
to DUCO cement and mix it in. You need to work fairly quickly as the DUCO
sets up with the graphite but the combination holds the sample to the stub
and has the conductivity necessary. I get the graphite from the vacuum
evaporator electrode sharpener. Colloidal graphite is not my friend!

-----Original Message-----
X-from: jpchandl-at-mines.edu [mailto:jpchandl-at-mines.edu]
Sent: Monday, April 22, 2013 10:38 AM
To: pekysar-at-ucdavis.edu

Stefan,

Those are good suggestions about fixation from Pat. You also might consider
the size and shape of your samples. If the bottom surface that you mount to
the stub is convex, so only the center is contacting the stub, you might not
be making adequate electrical contact between the upper coated surface and
the grounded stub. Sputter coating can have a difficult time getting into
the gap between the edge of the sample and the stub, and the coated surface
might not be grounded. One solution for this issue is to fill that gap with
something like colloidal silver paint or colloidal graphite (DAG). If you
coat after filling the gap, you can use something that is not conductive.

Good luck,

--John

John Chandler
EM Lab
Colorado School of Mines
jpchandl-at-mines.edu

On Apr 22, 2013, at 11:09 AM, {pekysar-at-ucdavis.edu}
wrote:

}
}
}
}
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}
} Hi Stephan,
} I've had good luck using the OTO method. Your times for your large plant
} tissue may differ and you may need to adjust accordingly. Take note that
the
} tannic acid and thiocarbohydrazide need to made up fresh. They may be
} buffered or not (I use aqueous).
} -Fix in primary fixative (some add Tannic to their Glut, I do not.
} -Rinse several times with buffer
} -Fix in 1% osmium for 1 hour
} -Immerse in either 1% tannic acid (TA) or saturated thiocarbohydrazide
(TCH)
} for at least 10 min-probably longer for large plant material
} -Rinse THOROUGHLY with ddH2O
} -Incubate in 1% Osmium tetroxide for 1 hour
} -Rinse thoroughly with ddH2O
} -You may stop here or repeat the last two steps, making sure to rinse
} THOROUGHLY.
} The TCH and TA are ligands which help the impregnation of osmium into
} tissues.
}
} Good Luck,
} Pat Kysar, C.E.M.T.
} Chair, Certification Board, Microscopy Society of America
} University of California, Davis
} School of Medicine
} Dept of Pathology and Laboratory Medicine
} Electron Microscopy Lab
}
}
} -----Original Message-----
} X-from: stefan.diller-at-t-online.de [mailto:stefan.diller-at-t-online.de]
} Sent: Monday, April 22, 2013 1:30 AM
} To: pekysar-at-ucdavis.edu
} Subject: [Microscopy] Osmiumtetroxide - Thiocarbohydrazide -
Osmiumteroxide
} Protocol
}
}
}
}
}
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}
} Dear All,
} anybody out to share a protocol for Osmiumtetroxide double fixing?
}
} I need it to hopefully get rid of the specimen charging at large plant
} specimen for SEM which did not work up to now either with
} "heavy" sputtercoating nor with an additional osmiumteroxide vapor
fixation
} step.
}
} Best regards,
} Stefan
}
}
} --
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 Phone
} ++49-931-7848701 Fax
} ++49-175-7177051 Mobile
}
} Websites:
} www.nanoflight.info
} www.stefan-diller.com
} www.electronmicroscopy.info
} www.elektronenmikroskopie.info
} www.zwillingsprojekt.de
} www.assisi.de
} Anfahrt: http://Mail.map24.com/Stefan.Diller
} -----------------------------------------------------
}
} ==============================Original
Headers==============================
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- Osmium teroxide Protocol
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11, 30 -- From jpchandl-at-mines.edu Mon Apr 22 12:28:01 2013
11, 30 -- Received: from EXCH-CAS4.adit.mines.edu (exch-cas4.Mines.EDU
[138.67.132.144])
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14.02.0342.003; Mon,
11, 30 -- 22 Apr 2013 11:28:01 -0600
11, 30 -- From: John Chandler {jpchandl-at-mines.edu}
11, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} ,
11, 30 -- Pat Kysar
11, 30 -- {pekysar-at-ucdavis.edu}
11, 30 -- Subject: Re: [Microscopy] RE: Osmium tetroxide -
Thiocarbohydrazide - Osmium
11, 30 -- teroxide Protocol
11, 30 -- Thread-Topic: [Microscopy] RE: Osmium tetroxide -
Thiocarbohydrazide -
11, 30 -- Osmium teroxide Protocol
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

In addition to John's sugestions for better grounding through sputter
coating, we also find that, if we have overhanging specimens, we can coat
from the sides as well. We lie the stubs on their sides in the coater (we
use pin stubs, so this works for us) and coat, then turn about a third of
a turn and coat, then another third of a turn and coat, followed by
a(nother) top coat if we think it's necessary. The side coatings do not
need to be especially thick, but they do help.

Aloha from rainy Honolulu (yay!),
Tina

} Those are good suggestions about fixation from Pat. You also might consider the size and shape of your samples. If the bottom surface that you mount to the stub is convex, so only the center is contacting the stub, you might not be making adequate electrical contact between the upper coated surface and the grounded stub. Sputter coating can have a difficult time getting into the gap between the edge of the sample and the stub, and the coated surface might not be grounded. One solution for this issue is to fill that gap with something like colloidal silver paint or colloidal graphite (DAG). If you coat after filling the gap, you can use something that is not conductive.
}
} Good luck,
}
} --John
}
} John Chandler
} EM Lab
} Colorado School of Mines
} jpchandl-at-mines.edu
}
}
}
} On Apr 22, 2013, at 11:09 AM, {pekysar-at-ucdavis.edu}
} wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Hi Stephan,
} } I've had good luck using the OTO method. Your times for your large plant
} } tissue may differ and you may need to adjust accordingly. Take note that the
} } tannic acid and thiocarbohydrazide need to made up fresh. They may be
} } buffered or not (I use aqueous).
} } -Fix in primary fixative (some add Tannic to their Glut, I do not.
} } -Rinse several times with buffer
} } -Fix in 1% osmium for 1 hour
} } -Immerse in either 1% tannic acid (TA) or saturated thiocarbohydrazide (TCH)
} } for at least 10 min-probably longer for large plant material
} } -Rinse THOROUGHLY with ddH2O
} } -Incubate in 1% Osmium tetroxide for 1 hour
} } -Rinse thoroughly with ddH2O
} } -You may stop here or repeat the last two steps, making sure to rinse
} } THOROUGHLY.
} } The TCH and TA are ligands which help the impregnation of osmium into
} } tissues.
} }
} } Good Luck,
} } Pat Kysar, C.E.M.T.
} } Chair, Certification Board, Microscopy Society of America
} } University of California, Davis
} } School of Medicine
} } Dept of Pathology and Laboratory Medicine
} } Electron Microscopy Lab
} }
} }
} } -----Original Message-----
} } X-from: stefan.diller-at-t-online.de [mailto:stefan.diller-at-t-online.de]
} } Sent: Monday, April 22, 2013 1:30 AM
} } To: pekysar-at-ucdavis.edu
} } Subject: [Microscopy] Osmiumtetroxide - Thiocarbohydrazide - Osmiumteroxide
} } Protocol
} }
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Dear All,
} } anybody out to share a protocol for Osmiumtetroxide double fixing?
} }
} } I need it to hopefully get rid of the specimen charging at large plant
} } specimen for SEM which did not work up to now either with
} } "heavy" sputtercoating nor with an additional osmiumteroxide vapor fixation
} } step.
} }
} } Best regards,
} } Stefan
} }
} }
} } --
} }
} }
} } -----------------------------------------------------
} } Stefan Diller - Scientific Photography
} } Arndtstrasse 22
} } D - 97072 Wuerzburg Germany
} } ++49-931-7848700 Phone
} } ++49-931-7848701 Fax
} } ++49-175-7177051 Mobile
} }
} } Websites:
} } www.nanoflight.info
} } www.stefan-diller.com
} } www.electronmicroscopy.info
} } www.elektronenmikroskopie.info
} } www.zwillingsprojekt.de
} } www.assisi.de
} } Anfahrt: http://Mail.map24.com/Stefan.Diller
} } -----------------------------------------------------
} }
} } ==============================Original Headers==============================
} } 8, 20 -- From stefan.diller-at-t-online.de Mon Apr 22 03:19:40 2013
} } 8, 20 -- Received: from mailout09.t-online.de (mailout09.t-online.de
} } [194.25.134.84])
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} } -0500
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} } 251.58]) by fwd53.t-online.de
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} } 8, 20 -- From: Stefan Diller {stefan.diller-at-t-online.de}
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} } Protocol
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} } 18, 30 -- From pekysar-at-ucdavis.edu Mon Apr 22 12:04:03 2013
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} } 18, 30 -- Mon, 22 Apr 2013 10:03:47 -0700
} } 18, 30 -- From: "Patricia Kysar" {pekysar-at-ucdavis.edu}
} } 18, 30 -- To: "Microscopy Listserver" {Microscopy-at-microscopy.com}
} } 18, 30 -- Cc: {stefan.diller-at-t-online.de}
} } 18, 30 -- Subject: RE: [Microscopy] Osmium tetroxide - Thiocarbohydrazide - Osmium teroxide Protocol
} } 18, 30 -- Date: Mon, 22 Apr 2013 10:03:30 -0700
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} 11, 30 -- From jpchandl-at-mines.edu Mon Apr 22 12:28:01 2013
} 11, 30 -- Received: from EXCH-CAS4.adit.mines.edu (exch-cas4.Mines.EDU [138.67.132.144])
} 11, 30 -- by ns.microscopy.com (8.12.11.20060308/8.12.8) with ESMTP id r3MHS18U020783
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} 11, 30 -- 22 Apr 2013 11:28:01 -0600
} 11, 30 -- From: John Chandler {jpchandl-at-mines.edu}
} 11, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com} ,
} 11, 30 -- Pat Kysar
} 11, 30 -- {pekysar-at-ucdavis.edu}
} 11, 30 -- Subject: Re: [Microscopy] RE: Osmium tetroxide - Thiocarbohydrazide - Osmium
} 11, 30 -- teroxide Protocol
} 11, 30 -- Thread-Topic: [Microscopy] RE: Osmium tetroxide - Thiocarbohydrazide -
} 11, 30 -- Osmium teroxide Protocol
} 11, 30 -- Thread-Index: AQHOP3wvjHZ2jGEl7EyU5C+YP2LliZji4tAA
} 11, 30 -- Date: Mon, 22 Apr 2013 17:28:00 +0000
} 11, 30 -- Message-ID: {E3AB3129-7237-40EB-856B-A4B6C8E64A0F-at-mines.edu}
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}

****************************************************************************
* 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*
****************************************************************************

==============================Original Headers==============================
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Dear All,
I will be teaching on an upcoming course at the headquarters of AFMWorkshop in Signal Hill, California. �The course will take place between July the 22nd and 26th, 2013.
The students will have access to AFMworkshop�s range of instruments, and we will also have a lot of test samples for experimenting. The course will be quite intensive, covering 5 mornings of lectures, and 5 afternoons of instrument use. We are restricting the course to a very small number of students, so you will get lots of coaching, and the course can be adapted to a certain extent to your interests. Both I and Paul West will be teaching, and the students will get a complimentary copy of our book, �Atomic Force Microscopy�. Although TT-AFM's from the AFMWorkshop will be used for the training, the course is open to users of all AFM systems.
�
Places are limited, and it has already begun to fill up, so I encourage anyone interested to book a place ASAP.
More details can be found at http://afmworkshop.com/afm-workshop.php.
Enquires, or registration can be made via email to info-at-afmworkshop.com.

____________________________________________________________________________
Atomic Force Microscopy
Dr Peter Eaton and Dr Paul West
Available through all good bookshops, or direct from Oxford University Press at:
http://ukcatalogue.oup.com/product/9780199570454.do http://afmhelp.com

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The LeBeau research group in the Department of Materials Science & Engineering at North Carolina State University is seeking a postdoctoral research associate to start July 2013. The post-doc will be responsible for the development and application of aberration corrected scanning transmission electron microscopy techniques and studying material interfaces, nanoparticles/nanocomposites, and device structures in a variety of systems including oxides, nitrides, and chalcogenides. These efforts will account for 60% of the post-doc's responsibilities.

The remaining 40% of the post-docs responsibilities will be to oversee training of students/researchers and research collaborations on the probe-corrected and monochromated FEI Titan G2 60-300 kV STEM/TEM microscope at NCSU. The post-doc will train users to operate the microscopy safely and independently, and will help develop facility policies for user training and oversight. The post-doc will also contribute to developing the internal and external user bases.

Eligible candidates will have a Ph.D. in Materials, Physical Sciences or related field and at least 3 years of experience in the theory and practice of scanning transmission electron microscopy. Candidates should also have experience with electronic measurement systems, programming, and a range of data analysis software packages. Strong interpersonal communication, commitment to collaborative research, and excellent time-management skills are essential. Interested candidates can apply at the following link: https://jobs.ncsu.edu/postings/22476

North Carolina State University is committed to affirmative action, equal opportunity and the diversity of its workforce.

James LeBeau
Assistant Professor
Department of Materials Science & Engineering
North Carolina State University
tel: (919) 515.5049
jmlebeau-at-ncsu.edu

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Hadeel,
I haven't seen any replies and I don't know if anyone is looking at beeswax
in an SEM, but you would have to start with a cold stage because the wax is
going to melt under the beam.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

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Name: hadeel

Title-Subject: [Filtered] sem of beeswax

Message: Hi Is it possible to study beeswax under the scanning electron
microscope and how they are
being studied under high vacuum

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Preliminary programme for the inaugral UBC 3D CLEM course, June 14-17, 2013

This course is designed for life science researchers/microscopists who are looking to learn 3D Correlative Microscopy.� Our instructors will guide you with a series of lectures and practical demonstrations.� We will introduce a sample (or two) at the beginning of the course and take that one sample through microCT, confocal/2P, dual beam FESEM and TEM tomography and into advanced 3D image processing.� Sample preparation will include both chemical fixation as well as high pressure freezing.

Synopsis:

When:�� June 14-17, 2013
Where:�� UBC Bioimaging Facility and UBC Centre for High-Throughput Phenogenomics, University of British Columbia, Vancouver, BC. Canada
Support:� Systems for Research (SFR), FEI Company, Edge Scientific, Leica Microsystems, Olympus Canada, TILL Photonics and VSG
Who:�� 10-12 students, preferably life sciences PhD students, Post docs, microscopy facility managers/directors/technicians.
Course fee: $2500 CDN (subject to change) and will include 5 nights accommodation, breakfast, lunch and one dinner.

For more information contact Garnet Martens at garnet.martens-at-botany.ubc.ca.

Preexisting knowledge of both light and electron microscopy necessary.

Lectures will include:

Principles of CLEM
CLEM methods
microCT for CLEM
Dual beam SEM for CLEM
Tomography
CryoEM sample preparation
Confocal/2P for CLEM
Image processing and analysis
Practical sessions:
microCT scanning
confocal/2p imaging
sample handling for both chemical fixation and high pressure freezing
FIB/SEM
Tomography
Image processing and analysis

Instructors:

Elizabeth Fischer, Rocky Mountain Labs, Hamilton, MT
Vinod Nair, Rocky Mountain Labs, Hamilton, MT
Bill Rice, New York Structual Biology Center
Kim Rensing, Leica Microsystems
Richard Gursky, FEI Company
Cliff Matheson, FEI Company
Christian Wietholt, VSG
TILL Photonics
Farid Jalali, Olympus Canada
Nancy Ford, UBC Centre for High-Throughput Phenogenomics
Gethin Owen, UBC Centre for High-Throughput Phenogenomics
Kevin Hodgson, UBC Bioimaging Facility
Derrick Horne, UBC Bioimaging Facility
Brad Ross, UBC Bioimaging Facility
Garnet Martens, UBC Bioimaging Facility

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11, 34 -- Subject: Announcing the inaugral UBC 3D CLEM course, June 14-17, 2013
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A colleague of mine would like to acquire a RHEED (Reflection
high-energy electron diffraction) system. I know this is not quite
microscopy � but I hope that it is close enough for the listserver. If
you know of a RHEED system that is no longer needed, please let me know.
Thank you for your help.

Please respond to me directly � not to the listserver.

Alwyn Eades
Department of Materials Science and Engineering
Lehigh University

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Dear Ken, dear Hadeel,
it is possible, with a trick in sputter-coating (I put a second chamber on the existing one, increasing the distance from target
to specimen and doing ca. 10 runs at very low current). I finally put the coated specimen into my high vac SEM and imaged at 5 kV.
See: http://www.elektronenmikroskopie.info/beegroup/wabe2_compo_m.htm
and
http://www.elektronenmikroskopie.info/beegroup/wabe3_06_c1.htm
I do not have a cold-stage...
The beam current might have been ca. 40 uA with LaB6.

Best wishes,
Stefan

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Am 24.04.13 22:11, schrieb kenconverse-at-qualityimages.biz:
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} Hadeel,
} I haven't seen any replies and I don't know if anyone is looking at beeswax
} in an SEM, but you would have to start with a cold stage because the wax is
} going to melt under the beam.
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Ravi,

What I did was just to pretend I was doing a negative stain for TEM,
without the negative stain.
Deposit the viri on a coated TEM grid, remove the fluid, a quick sputter
coater (Quick), or none. This was for a small dodecahedral virus that
attacked blue-green bacteria.
This is assuming you have a low-voltage FE-SEM. Fixation and a quick
coat might be more necessary if you are using a tungsten filament SEM.
Osmium might help, but I haven't tried it. OsO4 vapor fix should do.

It also helps if your phages are attached to their victim's flagellae or
pili. Makes a neater image, anyway.

Phil

} Email: ravi.thakkar369-at-gmail.com
} Name: Ravi Thakkar
}
} Organization: NIMHANS
}
} Title-Subject: [Filtered] How to process Bacteriophage for SEM ?
}
} Message: Dear Listener,
} How to process bacteriophage for SEM.?
} Is there osmium treatment will help in SEM imaging.?

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Email: ARD3A-at-virginia.edu
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Title-Subject: [Filtered] Ua/Pb stain process unknown debris

Message: Have suddenly beem plagued with debris after Ua/Pb staining of EMbed-812 embedded thin
sections. I am using the same proceedure I have used for years with no proplem. I have not seen
this type of debris except for one short period of time 3 years ago, never identified and clered up
after a few staining sessions. This time I have had the oroblem for weeks. I have reviewed all my
techniques for changes and replaced all reagents. Problem has no common demoninator. Debris is 1-2
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to the beam, reactive in the beam. Has anyone seem this and know what it is? Thx

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Email: jacqueline.ayotte-at-ticona.com
Name: Jackie Ayotte

Organization: Ticona Engineering Polymers

Title-Subject: [Filtered] Out of plane birefringence

Message: Dear Listers,

One of my internal customers is interested in measuring out of plane birefringence.
I am unsure as to what this is and have not been able to find useful information on the internet
as of yet. Can someone direct me to a resource or resources where I can learn about out of plane
birefringence and how to measure it?

Much Thanks,

Jackie

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We are trying to keep our 1995 Zeiss DSM 982 FESEM running for awhile longer. Unfortunately the viewing monitor has died, and we are having a hard time locating any compatible replacement monitors. The plate on the back says EGI Type FA 3435 L9 EHUL ZE0A, 230 V AC, 50 Hz. It has input plugs for R, G and B BNC connectors. I am told it contains a non-standard long persistence tube. If you have such a monitor sitting around in the back room, preferably in the New England area, I'd love to talk with you.

Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369

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Dear all,
dear Jackie,
good morning!

As to the circumstance that admittedly I haven't had knowledge about that matter but having interest also in all kinds of L-Microscopy I just tried to get some information from the net and found the following sources (if the URL's given display Company's websites: Disclaimer as usual: "no affiliation, no financial interest etc." :

http://www.google.com/patents/US20050219528 (also with construction sketches..)
citation: { The disclosure is directed to precise measurement of out-of-plane birefringence properties of samples of transparent optical material. Two angled-apart light beams are passed through a selected location of a sample optical element. One of the beams is incident to the sample surface. The characteristics of the beams are detected after passing through the sample, and the information detected is processed to determine the out-of-plane birefringence. } ...
citation: {..The prior art, including U.S. Pat. No. 6,473,747, Birefringence Measurement System, hereby incorporated by reference, discloses methods and apparatus for measuring birefringence of a sample using a light beam that is directed through the sample at a normal (zero-degree) incidence angle relative to the surface of the sample. As a result, the determination of the sample's birefringence is "in-plane," meaning that the determination essentially represents the difference between the indices of refraction of two orthogonal axes in a plane of the sample, that plane being normal to the incident light beam. ...}
citation: {..It is important to properly characterize the birefringence of such films, and other optical materials, in planes that are parallel to the normal (zero-degree) angle of incidence. This birefringence measure can be referred to as "vertical" or "out-of-plane" birefringence. One can consider the notion of in-plane and out-of-plane birefringence in terms of a Cartesian coordinate system. Accordingly, if the normal-incidence light is considered to travel in a direction parallel to the Z-axis of such a coordinate system, the in-plane birefringence occurs in the XY plane of the sample. Out-of-plane birefringence is in a plane perpendicular to the in-plane birefringence, thus occurring in the XZ or YZ plane.
Other applications (in additional to the birefringence compensation film example just discussed) may call for precise determination of out-of-plane birefringence. ... }
http://www.rdmag.com/articles/2004/07/how-measure-birefringence-ellipsometry
[http://en.wikipedia.org/wiki/Birefringence]: no term "out of plane/out-of-plane" found
http://polymer.homepcenter.co.kr/base/img/PDF_paper/2006-6.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3218218/: 2011: A quantitative collagen fibers orientation assessment using birefringence measurements: Calibration and application to human osteons.

http://onlinelibrary.wiley.com/doi/10.1889/1.1831012/abstract
Abstract 2004 (PPV= pay-per-view only) : P-55: A High Accuracy Instrument for Measuring Both In-Plane and Out-Of-Plane Birefringence in Birefringence Films

and more cf. GOOGLING: { out-of-plane birefringence out of plane }

Best wishes for a splendid and a recreative weekend,
best regards
Wolfgang MUSS
EM-Lab, Pathology
SALK-Salzburger Landeskliniken-LKH (Gen.Hospital)
SALZBURG-Austria

Personal Note: SCUR-SSSR Meeting SALZBURG 12-14th May 2013: have a short look into: http://orgs.dermis.net/scur/scur2013/content/e01home/e01home/index_ger.html
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Von: microscopylistserver-noreply-at-microscopy.com [mailto:microscopylistserver-noreply-at-microscopy.com]
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An: Mu� Wolfgang
Betreff: [Microscopy] Out of plane birefringence

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Dear Listers,

One of my internal customers is interested in measuring out of plane birefringence.
I am unsure as to what this is - and have not been able to find useful information on the internet
as of yet. Can someone direct me to a resource or resources where I can learn about out of plane
birefringence and how to measure it?

Much Thanks,

Jackie

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The Central Facility for Advanced Microscopy and Microanalysis (CFAMM) at the University of California at Riveside has an open position for electron microscopy laboratory technician. Position responsibilities include operation and routine maintenance of the electron microscopes and all phases of sample preparation and analysis for various types of materials. This position includes training users on specimen preparation, microscope operation, and related techniques.

To apply for the job visit: https://irecruitportal.ucr.edu/irecruit/!Controller?action=jobs_webui.show_page&page=jobs_browser&public=true&module=jobs.

//////////////////////////////////////////////////////////////////////////
Krassimir N Bozhilov, PhD
Central Facility for Advanced Microscopy and Microanalysis - CNAS
Materials Science and Engineering - BCOE
University of California
Riverside, CA 92521

ph. 951 827 2998
fax 951 827 2489
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Dear all,

if you have (pre-)registered for this meeting already, please register
again at http://nanocrystallography.net/ASCI2013 in order to receive
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arrangements / tax documentation / future reimbursements. This
personalized email will be the only "receipt" that we are going to
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If you are pondering attending this meeting, please check out
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The free two day workshop, "Advances in Structural and Chemical
Imaging", by the PREMIER* network on May 29 and 30, 2013, at CAMCOR,
University of Oregon, Eugene, OR (* Pooled Resources for Electron
Microscopy Informatics Education and Research). There are 22 invited
speakers and up to 25 posters. Space is limited and time flies, so
please register soon.

Scopes

1. structural analysis of organics, membrane proteins and macromolecules

2. structural analysis of inorganics and nanocrystals

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crystallography, surface reconstructions

4. full chemical analysis of crystalline phases by combining
atomic level structural and spectroscopic analyses

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and interface analyses at the nanometer length scale

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7. reconstruction algorithms for 3-D imaging and compressed sensing

We envision an interdisciplinary meeting that bridges the major divide
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communities. The multitude of methods being employed are linked by
common themes in their desire to achieve chemical specificity, high
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process and analyze large data sets to identify signature events.

Attached are the schedule (22 invited speakers), driving directions
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Required registration at http://nanocrystallography.net/ASCI2013.

You are invited to present posters, additional registrations for
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Vendors who would like to represent their companies are to register in
addition with Randy and Joe (smithran-at-pdx.edu,
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Northwest Section of the Microscopy Society of America.

The scopes of the meeting involve applications of the following methods:

- high-resolution transmission electron microscopy (TEM)

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spectroscopy (EELS), energy dispersive X-ray spectroscopy (EDXS), and
other spectroscopies

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- direct space tomography with STEM and TEM

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with EDXS in scanning electron microscopy (SEM)

- advanced electronic data management, telemicroscopy, open
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- low energy electron diffraction (LEED)

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- scanning tunneling microscopy (STM) and all kinds of
scanning probe microscopies (SPM)

- X-ray microscopy, tomography and spectroscopy

- advanced light microscopy

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--
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Associate Professor of Physics

Nano-Crystallography Group:
http://nanocrystallography.research.pdx.edu/nano-crystallography-group.
Free (as in free speech rather than free beer) crystallographic
information and interactive 3D visualizations of crystal structures
and morphologies with educational relevance:
http://nanocrystallography.research.pdx.edu/databases.

Portland State University
Department of Physics
P.O. Box 751, Portland, OR 97207-0751
Tel.: 503 725 4227, Departmental Fax.: 503 725 2815
pmoeck-at-pdx.edu
www.physics.pdx.edu/~pmoeck

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Office hours during quarters Tuesdays and Thursdays 12:00 to 12:30 and
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for more interactive 3D display options try:
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Webportal: Open Access Crystallography Resource Portal
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Hi Art

I'm not sure what your technique involves but we had a contaminant issue arising from the filter paper that we used to push the grids from the forceps after washing.

Regards
Naomi

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Email: ARD3A-at-virginia.edu
Name: Art Dewey

Organization: Univ. of Va Health Center /Pathology

Title-Subject: [Filtered] Ua/Pb stain process unknown debris

Message: Have suddenly beem plagued with debris after Ua/Pb staining of EMbed-812 embedded thin
sections. I am using the same proceedure I have used for years with no proplem. I have not seen
this type of debris except for one short period of time 3 years ago, never identified and clered up
after a few staining sessions. This time I have had the oroblem for weeks. I have reviewed all my
techniques for changes and replaced all reagents. Problem has no common demoninator. Debris is 1-2
um to 20 um in size, light grey to dark grey, semi-circular to half circle/wedge shape, trans-lucent
to the beam, reactive in the beam. Has anyone seem this and know what it is? Thx

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I had a question on an exam I gave recently and now I am trying to figure out what should be a correct answer. The question was whether you could tell an actin filament (8nm) from a bundle of filaments by fluorescence microscopy. They were supposed to think about image resolution and brightness in their answer. Obviously, you would be unlikely to be able to resolve filaments in a bundle since they are so tightly packed. But a bundle should be much brighter than a filament if you had both to compare.
My question is, at what point would the width of the structure begin to give you information about the size of the bundle? To make things simple, lets assume the filaments were in a parallel, flat array with the 8nm filaments spaced by a cross-linking protein like alpha-actinin (about 35nm long). If your image resolution was 300nm, when would the image of a fluorescent bundle look "wider" than the image of a single filament? Thanks- Dave

David Knecht, Ph.D.
Professor and Head of Microscopy Facility
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

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David,
Imagine your specimen is the world's best 'nano-iris'
surrounded by complete dark sample. Let the diameter of the iris be
anything, down all the way to zero. Start out with the iris at
something absurdly tiny, say 1 nm (and pretend/assume that enough
light can be tranmitted through to form an image). Your imaging
system will image this point as an Airy disc, whose diameter is set
by the optics. Now open the iris. As you do, the Airy disk will get
brighter, but its size will not change until the iris diameter
becomes greater than that limiting Airy disc diameter.

Then to see how this works in resolution, you need to imagine
two ! of these gems side by side on the slide, and translocate one
away from the other. Again, once the distance between them exceeds
about half of the Airy disc diameter, you have a chance of seeing
them as two things, not as one. Hence resolution. The various
different formulae reflect the fact that there is no obvious way to
define whether you have two Airy discs or just one. But they all
converge to a distance that amounts to about one half a wavelength
(of the incident light).

That is in terms of size. You will note that having two iris
together will let more light through and so the fused (unresolved
spatially) pair of irises will be brighter than just one. So if you
know for sure how much light one iris gets through (with a fixed
diameter), you can tell by that criterion, if you have one, two or
more within an Airy-disc sized spot. To the extent that each actin
filament has a constant amount of fluorochrome bound, then this kind
of intensity counting can in principle be used to count the number of
filaments, even when they cannot be seen as objects. (practical
matters like quenching and so on are a different story!).

Anyway this is how I have always understood it.

As ever
Tobias

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Hi David:

I think the question, or how the question is being asked, is as important as the answer.

IF the question is "whether you could tell an actin filament (8nm) from a bundle of filaments by fluorescence microscopy", the answer should be "NO", regardless of the brightness, simply because of the resolution limit.

IF the question is "whether you could 'see' a single actin filament(8nm), using a 200nm resolution microscope", the answer is "YES", as long as the filament has "enough contrast". It has nothing to do with resolution.

IF the question is "whether you could tell if that is a single actin filament, or actin bundle (more than 1 filaments)", the answer is "MAYBE", if you could provide a reference image of a single filament, and assume all filaments have the same brightness.

Zhaojie Zhang, Ph. D.
Director, Jenkins Microscopy Facility
University of Wyoming
Laramie, WY 82071

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I had a question on an exam I gave recently and now I am trying to figure out what should be a correct answer. The question was whether you could tell an actin filament (8nm) from a bundle of filaments by fluorescence microscopy. They were supposed to think about image resolution and brightness in their answer. Obviously, you would be unlikely to be able to resolve filaments in a bundle since they are so tightly packed. But a bundle should be much brighter than a filament if you had both to compare.
My question is, at what point would the width of the structure begin to give you information about the size of the bundle? To make things simple, lets assume the filaments were in a parallel, flat array with the 8nm filaments spaced by a cross-linking protein like alpha-actinin (about 35nm long). If your image resolution was 300nm, when would the image of a fluorescent bundle look "wider" than the image of a single filament? Thanks- Dave

David Knecht, Ph.D.
Professor and Head of Microscopy Facility Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

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I asked several months ago.....but I just got back around to this:
Any Porter Blum MT2 maint. repair parts- belts(orings?) and motor mounts Suppliers out there? any DIYers?

You can just email me: wa5ekh-at-juno.com Thanks!

jeff/ Dallas Area
____________________________________________________________
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Get the latest details on the new BlackBerry 10 smartphone.
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Scripting School At Microscopy and Microanalysis 2013; 1:30 pm - 5:30 pm,
Sunday, August 4, 2013; Indiana Convention Center, Room 233, 2nd Floor;
Indianapolis, IN, USA. Complimentary to all registered participants.

Just prior to the opening of M&M 2013, by popular demand, Gatan will again
run a school on the basics of Scripting within Gatan DigitalMicrograph
(DM) software. The School is intended for everyone who would like to
learn about the capabilities and use of DM Scripting and is suitable for
beginners, occasional users or anyone wanting a refresher. As it will be a
mixture of talks and practical examples using DM Scripting, participants
should bring their own laptop computer running Gatan Microscopy Suite
(GMS) version 2. Afternoon refreshments will be provided and the event is
timed to lead into the M&M Opening, in the same location at 6.30pm. This
event is free of charge. Registration is required. Please note that
demonstration offline DM imaging licenses are available at www.gatan.com.
To download DigitalMicrograph (DM) offline 32 or 64 bit licenses, please
go to: http://www.gatan.com/products/software/softwaredemo.php
Registration: For online registration, please go to:
http://www.gatan.com/products/software/DMScriptingSchoolMM13RegForm.php.
Space is limited. Please submit your online registration by July 5, 2013.
For more information on this event, please email Gatan at: info-at-gatan.com.
Gatan reserves the right to cancel any Gatan school due to low enrollment
or for any other reason, at any time.

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5, 25 -- From: John Hyun {JHyun-at-gatan.com}
5, 25 -- To: Microscopy Server {microscopy-at-microscopy.com}
5, 25 -- Subject: Re: Scripting School at M&M 2013
5, 25 -- Thread-Topic: Scripting School at M&M 2013
5, 25 -- Thread-Index: AQHORfYVRrN1FIoD+0KlKoEzTld/3w==
5, 25 -- Date: Tue, 30 Apr 2013 22:57:26 +0000
5, 25 -- Message-ID: {CDA59958.24916%jhyun-at-gatan.com}
5, 25 -- In-Reply-To: {CDA598C7.24911%jhyun-at-gatan.com}
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Gatan wants to see the best of your TEM images, taken on one of our camera
or energy filter systems. Gatan is running a competition with iPad/Apple
prizes* for the images judged best in five categories: life science,
material science, diffraction pattern,
energy filtered TEM and artistic merit. To be eligible, the image must be
approved for open publication. The best images will be shown on the Gatan
website, in Gatan publications and elsewhere. Our imaging R&D experts will
be the judges for this competition.

Winners will be announced in July and prizes presented at M&M 2013
(Indianapolis) or sent electronically. To register online please go to:
http://www.gatan.com/company/news/news03111301.php

Image Submission: Once you register online, you will receive a
confirmation email from Gatan with complete instructions on how to submit
your image file, including Gatan DigitalMicrograph� image format, upload
instructions and ftp site information
for large files. There is no limit on the number of entries per person.
The deadline for all entries is June 30, 2013. Good luck and we look
forward to seeing your images!

*Winners will receive an Apple e-gift card for 1,000 US Dollars or their
local currency equivalent. The Apple name and iPad are registered
trademarks of Apple Inc.
--

==============================Original Headers==============================
6, 25 -- From JHyun-at-gatan.com Tue Apr 30 17:58:33 2013
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6, 25 -- From: John Hyun {JHyun-at-gatan.com}
6, 25 -- To: Microscopy Server {microscopy-at-microscopy.com}
6, 25 -- Subject: Image competition for Apple prizes
6, 25 -- Thread-Topic: Image competition for Apple prizes
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Hello all,
I am looking into TEM of polymers and nanocomposites. This is
rather different from my background of semiconductor materials,
functional oxides and all that other hard stuff. I'm more or less happy
with the TEM side of things, low dose is just another technique to learn
- but I would like any advice you can give me on specimen prep. So -
what kit and expertise would you expect to find in a good TEM lab
dealing with soft matter? Who are the main suppliers I should be
talking to? And anything else which you think it would be Good To Know?

Thanks a lot

Richard

--
Richard Beanland

Director,
Integrity Scientific Ltd.

==============================Original Headers==============================
6, 21 -- From contact-at-integrityscientific.com Wed May 1 02:30:45 2013
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Dear List,

We have three ion getter pumps from Philips TEMs that I would like to
scrap or have recycled (I don't need them to be refurbished and returned).

Has someone (preferably located in the UK) done this before? Do they have
any value?

Best regards,

Ben

--
Research Support Manager
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271649
{http://www.mrc.ox.ac.uk/}

==============================Original Headers==============================
11, 30 -- From ben.micklem-at-pharm.ox.ac.uk Wed May 1 04:55:51 2013
11, 30 -- Received: from relay13.mail.ox.ac.uk (relay13.mail.ox.ac.uk [129.67.1.166])
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11, 30 -- From: Ben Micklem {ben.micklem-at-pharm.ox.ac.uk}
11, 30 -- To: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
11, 30 -- Subject: IGP scrapping/recycling
11, 30 -- Thread-Topic: IGP scrapping/recycling
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Dear all,

Announcing a full day meeting of the Microscopy Society of Northeastern
Ohio joint with the Fifty-Seventh Annual SAS/ACS/AVS May Conference on
Wednesday, May 15th, 2013.

Join us for this full day meeting held at the Dolan Science Center at John
Carroll University as we once again partner with SAS/ACS/AVS in a similar
format to the previous two years.

Additional information can be found at: http://www.msneo.org/index.html

Thank you,

Chris Holp
Past President, MSNO

-----------------------------------------
The information contained in this message is intended only for the
personal and confidential use of the recipient(s) named above. If
the reader of this message is not the intended recipient or an
agent responsible for delivering it to the intended recipient, you
are hereby notified that you have received this document in error
and that any review, dissemination, distribution, or copying of
this message is strictly prohibited. If you have received this
communication in error, please notify us immediately, and delete
the original message.

==============================Original Headers==============================
9, 31 -- From holpc-at-firstenergycorp.com Wed May 1 10:26:45 2013
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Ian Holton
Acutance Scientific

-----Original Message-----
X-from: ben.micklem-at-pharm.ox.ac.uk [mailto:ben.micklem-at-pharm.ox.ac.uk]
Sent: 01 May 2013 11:08
To: ian-at-acutance.co.uk

Dear List,

We have three ion getter pumps from Philips TEMs that I would like to scrap
or have recycled (I don't need them to be refurbished and returned).

Has someone (preferably located in the UK) done this before? Do they have
any value?

Best regards,

Ben

--
Research Support Manager
MRC Anatomical Neuropharmacology Unit, Mansfield Road, Oxford, OX1 3TH,
United Kingdom. Telephone: 01865 271649
{http://www.mrc.ox.ac.uk/}

==============================Original Headers==============================
11, 30 -- From ben.micklem-at-pharm.ox.ac.uk Wed May 1 04:55:51 2013 11, 30 --
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-0500
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Wed, 1 11, 30 -- May 2013 10:55:49 +0100 11, 30 -- From: Ben Micklem
{ben.micklem-at-pharm.ox.ac.uk} 11, 30 -- To: "microscopy-at-microscopy.com"
{microscopy-at-microscopy.com} 11, 30 -- Subject: IGP scrapping/recycling 11,
30 -- Thread-Topic: IGP scrapping/recycling 11, 30 -- Thread-Index:
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21, 24 -- From ian-at-acutance.co.uk Wed May 1 12:01:00 2013
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21, 24 -- From: "Ian Holton" {ian-at-acutance.co.uk}
21, 24 -- To: {ben.micklem-at-pharm.ox.ac.uk} , {Microscopy-at-microscopy.com}
21, 24 -- References: {201305011008.r41A8AjJ012712-at-ns.microscopy.com}
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If you haven't seen IBM's 60 second movie "A boy and his atom" made using a scanning tunneling microscope, Google the title and there are a lot of links out there. I didn't paste one here because I suspected the listserver's filters would have blocked it. They used 242 frames of stop motion photography at 100 million x magnification to create this cute movie. Enjoy. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

==============================Original Headers==============================
6, 28 -- From PhillipsT-at-missouri.edu Wed May 1 17:59:52 2013
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6, 28 -- Subject: A boy and his atom
6, 28 -- Thread-Topic: A boy and his atom
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The 14th Frontiers of Electron Microscopy in Materials Science (FEMMS)
meeting will be held in the picturesque town of Lorne on the spectacular
Great Ocean Road of Victoria, Australia.

The meeting will run from 8th - 13th September. Over 35 invited
speakers, showcasing the best in new electron microscopy, will present
in this single session conference.

Registration is open, and abstracts are being accepted for posters.

For more information visit the website at http://www.femms2013.org/

--
Dr M.Weyland, FEMMS 2013 Chair
--------------------------------------------------------------------------
Monash Centre for Electron Microscopy
Monash University
Victoria, Australia
--------------------------------------------------------------------------
www.mcem.monash.edu.au
-------------------------------------------------------------------------

==============================Original Headers==============================
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Name: Amit Gupta

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Title-Subject: [Filtered] Silicon standard sample for tem

Message: Good evening everyone,
We need a silicon sample ,with thickness down to atomic level, and oriented along 111 direction.
Only commercially sample available is Mag*i*cal but its just too costly!!
I was wondering if there is some easy way to prepare it in lab (we have silicon crystal), ie
1) Is there any way to tell 111 orientation in bulk crystal?( like some cleave plain etc)
2) can we do it "chemically"? As ours is a chemistry lab it would be lot more easier for us then ion
milling etc (like etching W tips for stm)

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For more information visit: http://manoa.hawaii.edu/chem/index.php?id=162

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Hi everybody,

it's http://www.guardian.co.uk/technology/video/2013/may/01/ibm-smallest-ever-animation-molecular-video

but why are there always two dots moving around when it should be only
one atom??? without a good explanation for that I am reluctant to show
it in my introductory nano-materials science and engineering class

kind regards, Peter Moeck

On Wed, May 1, 2013 at 4:15 PM, {PhillipsT-at-missouri.edu} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} If you haven't seen IBM's 60 second movie "A boy and his atom" made using a scanning tunneling microscope, Google the title and there are a lot of links out there. I didn't paste one here because I suspected the listserver's filters would have blocked it. They used 242 frames of stop motion photography at 100 million x magnification to create this cute movie. Enjoy. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://biology.missouri.edu/people/?person=88
} http://www.biotech.missouri.edu/mcc/
}

--
Peter Moeck, PhD (Dr. rer. nat. Crystallography)
Associate Professor of Physics

Nano-Crystallography Group:
http://nanocrystallography.research.pdx.edu/nano-crystallography-group.
Free (as in free speech rather than free beer) crystallographic
information and interactive 3D visualizations of crystal structures
and morphologies with educational relevance:
http://nanocrystallography.research.pdx.edu/databases.

Portland State University
Department of Physics
P.O. Box 751, Portland, OR 97207-0751
Tel.: 503 725 4227, Departmental Fax.: 503 725 2815
pmoeck-at-pdx.edu
www.physics.pdx.edu/~pmoeck

Office 404, Science Research and Teaching Center
1719 SW 10th Ave, Portland, OR 97201

Office hours during quarters Tuesdays and Thursdays 12:00 to 12:30 and
per appointment

North American mirror of the Crystallography Open Database, COD,
http://nanocrystallography.org, with more than 210,000 carefully
evaluated datasets for download and interactive visualization in 3D,
for more interactive 3D display options try:
http://nanocrystallography.research.pdx.edu/search/codmirror/
Webportal: Open Access Crystallography Resource Portal
http://nanocrystallography.net.

"... no hell below us, above us only sky ..." John Lennon
"... der wackere Schwabe forcht sich nit ..." Albert Einstein
(inspired by Ludwig Uhland's poem)
"Was auch immer geschieht: Nie duerft ihr so tief sinken, von dem
Kakao, durch den man euch zieht, auch noch zu trinken." Erich Kaestner
"... the only thing needed for a happy life is a project one is
enthusiastic about." John C. H. Spence, Acta Cryst. A 69 (2013) 25-33

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The version on youtube has a link to a "behind the scenes" movie.

http://www.youtube.com/watch?v=xA4QWwaweWA&feature=youtu.be

About 1:41 in they mention that the man is built with carbon monoxide molecules, which would explain the two atoms.

So the title should really be "a boy and his molecule"

TobyS

Dr Tobias Starborg
Senior Experimental Officer: 3DEM
Wellcome Centre for Cell Matrix Research
Michael Smith Building
Manchester
M13 9PT
Tel: +44(0)1612755170
http://manchesterpolara.org.uk

________________________________________
X-from: pmoeck-at-pdx.edu [pmoeck-at-pdx.edu]
Sent: 02 May 2013 07:38
To: Tobias Starborg

Hi everybody,

it's http://www.guardian.co.uk/technology/video/2013/may/01/ibm-smallest-ever-animation-molecular-video

but why are there always two dots moving around when it should be only
one atom??? without a good explanation for that I am reluctant to show
it in my introductory nano-materials science and engineering class

kind regards, Peter Moeck

On Wed, May 1, 2013 at 4:15 PM, {PhillipsT-at-missouri.edu} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} If you haven't seen IBM's 60 second movie "A boy and his atom" made using a scanning tunneling microscope, Google the title and there are a lot of links out there. I didn't paste one here because I suspected the listserver's filters would have blocked it. They used 242 frames of stop motion photography at 100 million x magnification to create this cute movie. Enjoy. Tom
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
} phillipst-at-missouri.edu
}
} http://biology.missouri.edu/people/?person=88
} http://www.biotech.missouri.edu/mcc/
}

--
Peter Moeck, PhD (Dr. rer. nat. Crystallography)
Associate Professor of Physics

Nano-Crystallography Group:
http://nanocrystallography.research.pdx.edu/nano-crystallography-group.
Free (as in free speech rather than free beer) crystallographic
information and interactive 3D visualizations of crystal structures
and morphologies with educational relevance:
http://nanocrystallography.research.pdx.edu/databases.

Portland State University
Department of Physics
P.O. Box 751, Portland, OR 97207-0751
Tel.: 503 725 4227, Departmental Fax.: 503 725 2815
pmoeck-at-pdx.edu
www.physics.pdx.edu/~pmoeck

Office 404, Science Research and Teaching Center
1719 SW 10th Ave, Portland, OR 97201

Office hours during quarters Tuesdays and Thursdays 12:00 to 12:30 and
per appointment

North American mirror of the Crystallography Open Database, COD,
http://nanocrystallography.org, with more than 210,000 carefully
evaluated datasets for download and interactive visualization in 3D,
for more interactive 3D display options try:
http://nanocrystallography.research.pdx.edu/search/codmirror/
Webportal: Open Access Crystallography Resource Portal
http://nanocrystallography.net.

"... no hell below us, above us only sky ..." John Lennon
"... der wackere Schwabe forcht sich nit ..." Albert Einstein
(inspired by Ludwig Uhland's poem)
"Was auch immer geschieht: Nie duerft ihr so tief sinken, von dem
Kakao, durch den man euch zieht, auch noch zu trinken." Erich Kaestner
"... the only thing needed for a happy life is a project one is
enthusiastic about." John C. H. Spence, Acta Cryst. A 69 (2013) 25-33

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Yes, there are two spots. Yes, it is a molecule, not an atom. But why
let that take away from the beauty of what is shown, or recognizing the
tremendous technological advances in imaging over the past 10 years.
It is a Boy and his Atom, and I watch it with overwhelming awe and
thanks that after 44 years in electron microscopy I am still privileged
enough to see such things.

And Tom, thanks for posting it.

paul
--
Paul R. Hazelton, PhD
Viral Gastroenteritis Study Group
University of Manitoba
Department of Medical Microbiology
511 Basic Medical Sciences Building
745 William Avenue
Winnipeg, Manitoba, Canada, R3E 0J9
e-mail: paul_hazelton-at-umanitoba.ca
paulhazelton-at-mts.net
Phone: 204-789-3313 (w);
204-489-6924 (h)
Cell: 204-781-6982
Fax: 204-789-3926

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ealname - Valerie Lecomte
Email - valerie.lecomte-at-usherbrooke.ca
ORGANIZATION - Sherbrooke University
EDUCATION - Graduate College
LOCATION - Chicoutimi, Quebec, Canada
SUBJECT_OF_QUESTION - SEM analysis of trace elements in humic soil
QUESTION - Hi,

I am studying environmental sciences at Sherbrooke University. I am
starting a new project that will include structural and chemical SEM
analysis (trace metals and carbon if possible) in soil samples mainly
consisted of organic matter (lots of fiber and few minerals).

I was wondering if you have a sample preparation method (stub type,
embedding and coating) to suggest me that would suit the type of
analysis I want to do.

Thank you very much for your precious help!

Valerie Lecomte

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Asylum Research, an Oxford Instruments company, will host a new webinar on May 22 � �Getting Started with AFM in Biology � It�s Easier Than You Think�. Asylum Applications Scientist Dr. Ir�ne Revenko, who has over 19 years of teaching and training AFM in biology, will be presenting and taking questions. The webinar is ideal for biologists that are new to AFM or for AFM experts starting to study biology.

The webinar will cover topics that are critical for successful AFM imaging including sample preparation, choosing the correct cantilever, imaging in fluids and choosing the correct measurement mode. These will be presented in the context of four case studies of typical biological samples:

1. Imaging DNA in liquid � including routine helix resolution
2. Imaging living cells in liquid
3. Measuring Young�s modulus of living cells
4. Unfolding forces in Titin

Registration for the 8:00am session is at https://www3.gotomeeting.com/register/702661318

Registration for the 4:00pm session is at https://www3.gotomeeting.com/register/707007462

Attendance will be limited, so early registration is recommended. For a complete listing of Asylum webinars, go to http://www.asylumresearch.com/Webinars/index.shtml

___________________________________________________________________________This e-mail is confidential and is for the addressee only. Please refer to www.oxinst.com/email-statement for regulatory information.

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I am not getting much response, so let me try to explain my interpretation of the problem. If you consider a single filament, then you expect to image a linear extension of the diffraction pattern (airy disk) of each fluorescent point source along the filament projected onto the chip. As a diffraction pattern, the emission will decrease stochasitically as you move away from the filament. In theory, if you wait long enough and acquire enough photons over a long time, the diffraction pattern spreads to decreasingly represented orders of the pattern. So what you call the "edge" of the filament as you image it is arbitrary and determined by the sensitivity of your detector and the imaging conditions. Now as you increase the number of filaments in a bundle, the total fluorescence output from the volume increases. If you were to acquire an image with the same gain and integration as you did for the single filament, you would see a "wider" structure because the higher orders of the pattern now become more readily visible due to the brighter signal and more photons coming from the same "sub-resolution" volume. If you normalize for the total fluorescence, I would expect the single filaments to look just like the bundles up to the point where you exceed optical resolution (about 8 filaments when you are nearing 300nm wide). Of course a real bundle is much more tightly packed as a cylinder, so it would take many more filaments to look larger. But if you do not normalize, I think the bundle will look larger than a single filament before you exceed the resolution limit. My greater point is that the imaging conditions and detector will have an effect on what you call the "size" of an object and this is a different set of considerations from that we normally think about as resolving of multiple objects from each other. Dave

On Apr 29, 2013, at 2:28 PM, david.knecht-at-uconn.edu {mailto:david.knecht-at-uconn.edu} wrote:

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I had a question on an exam I gave recently and now I am trying to figure out what should be a correct answer. The question was whether you could tell an actin filament (8nm) from a bundle of filaments by fluorescence microscopy. They were supposed to think about image resolution and brightness in their answer. Obviously, you would be unlikely to be able to resolve filaments in a bundle since they are so tightly packed. But a bundle should be much brighter than a filament if you had both to compare.
My question is, at what point would the width of the structure begin to give you information about the size of the bundle? To make things simple, lets assume the filaments were in a parallel, flat array with the 8nm filaments spaced by a cross-linking protein like alpha-actinin (about 35nm long). If your image resolution was 300nm, when would the image of a fluorescent bundle look "wider" than the image of a single filament? Thanks- Dave

David Knecht, Ph.D.
Professor and Head of Microscopy Facility
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

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David Knecht, Ph.D.
Professor and Head of Microscopy Facility
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

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Name: Patrick McCurdy

Organization: Colorado State University

Title-Subject: [Filtered] Materials TEM position

Message: The position of Electron Microscopist and Imaging Specialist is a full-time, special, 12
month Research Scientist/Scholar I administrative professional position in the CSU Chemistry
Departments Central Instrument Facility (CIF) Imaging and X-ray Spectroscopy laboratories.
Responsibilities will include providing advanced research services and training for, and routine
maintenance of, electron microscopy methods (EM) including ultra-high resolution transmission EM.
All instruments in the CIF are available for use by the successful candidate,who will be expected to
collaborate with colleagues, staff scientists, faculty and other principal investigators to
advance CSUs primary research, education and service mission. Additional information about the CIF
can be found at http://cif.colostate.edu/. The successful candidate must have Ph.D. in a
chemistry-related discipline. Familiarity with UHV, EM and X-ray techniques is required and
development of collaborations with senior investigators is expected. Familiarity with the chemistry
of hard materials, electron energy loss spectroscopy, lithography and focused ion-beam milling will
be considered advantageous. Salary is negotiable and will be commensurate with experience.

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Hi David,

I'm not sure I can follow your argument, but I'd like to point out some things
that seem a bit unclear to me.
You say that the Airy disk is the diffraction pattern of a point source.
As I understand it the Airy disk is the real space image of a point source.
The ideal image of a point source would be a point, but due to the limited size of
the lenses this point is convoluted with the airy disk, which is the diffraction pattern
of the aperture (edge of the lens).
The diffraction pattern of a point source should be a constant (just like the Fourier
transform of a delta function).
Maybe your argument is perfectly alright, if you reformulate it and clarify whether you
are considering a real space image or a diffraction pattern (in Fourier space).

Hope this helps,

Philip

-----Original Message-----
X-from: david.knecht-at-uconn.edu [mailto:david.knecht-at-uconn.edu]
Sent: 03 May 2013 00:44
To: Philip K�ck

I am not getting much response, so let me try to explain my interpretation of the problem. If you consider a single filament, then you expect to image a linear extension of the diffraction pattern (airy disk) of each fluorescent point source along the filament projected onto the chip. As a diffraction pattern, the emission will decrease stochasitically as you move away from the filament. In theory, if you wait long enough and acquire enough photons over a long time, the diffraction pattern spreads to decreasingly represented orders of the pattern. So what you call the "edge" of the filament as you image it is arbitrary and determined by the sensitivity of your detector and the imaging conditions. Now as you increase the number of filaments in a bundle, the total fluorescence output from the volume increases. If you were to acquire an image with the same gain and integration as you did for the single filament, you would see a "wider" structure because the higher orders !
of the pattern now become more readily visible due to the brighter signal and more photons coming from the same "sub-resolution" volume. If you normalize for the total fluorescence, I would expect the single filaments to look just like the bundles up to the point where you exceed optical resolution (about 8 filaments when you are nearing 300nm wide). Of course a real bundle is much more tightly packed as a cylinder, so it would take many more filaments to look larger. But if you do not normalize, I think the bundle will look larger than a single filament before you exceed the resolution limit. My greater point is that the imaging conditions and detector will have an effect on what you call the "size" of an object and this is a different set of considerations from that we normally think about as resolving of multiple objects from each other. Dave

On Apr 29, 2013, at 2:28 PM, david.knecht-at-uconn.edu {mailto:david.knecht-at-uconn.edu} wrote:

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I had a question on an exam I gave recently and now I am trying to figure out what should be a correct answer. The question was whether you could tell an actin filament (8nm) from a bundle of filaments by fluorescence microscopy. They were supposed to think about image resolution and brightness in their answer. Obviously, you would be unlikely to be able to resolve filaments in a bundle since they are so tightly packed. But a bundle should be much brighter than a filament if you had both to compare.
My question is, at what point would the width of the structure begin to give you information about the size of the bundle? To make things simple, lets assume the filaments were in a parallel, flat array with the 8nm filaments spaced by a cross-linking protein like alpha-actinin (about 35nm long). If your image resolution was 300nm, when would the image of a fluorescent bundle look "wider" than the image of a single filament? Thanks- Dave

David Knecht, Ph.D.
Professor and Head of Microscopy Facility Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

==============================Original Headers==============================
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Hi David,
�

the diffraction limit is no more a limit nowadays.�Recent techniques of flurescence microscopy appeared which allow resolutions far under the diffraction limit.
STED from Stefan Hell is on of them.
Just to point out this fact.

Regards,
Stephane

----- Original Message -----
X-from: "Philip.Koeck-at-ki.se" {Philip.Koeck-at-ki.se}
To: nizets2-at-yahoo.com
Cc:
Sent: Friday, May 3, 2013 9:27 AM

Hi David,

I'm not sure I can follow your argument, but I'd like to point out some things
that seem a bit unclear to me.
You say that the Airy disk is the diffraction pattern of a point source.
As I understand it the Airy disk is the real space image of a point source.
The ideal image of a point source would be a point, but due to the limited size of
the lenses this point is convoluted with the airy disk, which is the diffraction pattern
of the aperture (edge of the lens).
The diffraction pattern of a point source should be a constant (just like the Fourier
transform of a delta function).
Maybe your argument is perfectly alright, if you reformulate it and clarify whether you
are considering a real space image or a diffraction pattern (in Fourier space).

Hope this helps,

Philip

-----Original Message-----
X-from: david.knecht-at-uconn.edu [mailto:david.knecht-at-uconn.edu]
Sent: 03 May 2013 00:44
To: Philip K�ck

I am not getting much response, so let me try to explain my interpretation of the problem.� If you consider a single filament, then you expect to image a linear extension of the diffraction pattern (airy disk) of each fluorescent point source along the filament projected onto the chip.� As a diffraction pattern, the emission will decrease stochasitically as you move away from the filament.� In theory, if you wait long enough and acquire enough photons over a long time, the diffraction pattern spreads to decreasingly represented orders of the pattern.� So what you call the "edge" of the filament as you image it is arbitrary and determined by the sensitivity of your detector and the imaging conditions.� Now as you increase the number of filaments in a bundle, the total fluorescence output from the volume increases.� If you were to acquire an image with the same gain and integration as you did for the single filament, you would see a "wider" structure
because the higher orders !
of the pattern now become more readily visible due to the brighter signal and more photons coming from the same "sub-resolution" volume.� If you normalize for the total fluorescence, I would expect the single filaments to look just like the bundles up to the point where you exceed optical resolution (about 8 filaments when you are nearing 300nm wide).� Of course a real bundle is much more tightly packed as a cylinder, so it would take many more filaments to look larger.� But if you do not normalize, I think the bundle will look larger than a single filament before you exceed the resolution limit.� My greater point is that the imaging conditions and detector will have an effect on what you call the "size" of an object and this is a different set of considerations from that we normally think about as resolving of multiple objects from each other.� Dave

On Apr 29, 2013, at 2:28 PM, david.knecht-at-uconn.edu {mailto:david.knecht-at-uconn.edu} wrote:

----------------------------------------------------------------------------
The Microscopy ListServer -- CoSponsor:� The Microscopy Society of America To� Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver

I had a question on an exam I gave recently and now I am trying to figure out what should be a correct answer.� The question was whether you could tell an actin filament (8nm) from a bundle of filaments by fluorescence microscopy.� They were supposed to think about image resolution and brightness in their answer.� Obviously, you would be unlikely to be able to resolve filaments in a bundle since they are so tightly packed.� But a bundle should be much brighter than a filament if you had both to compare.
My question is, at what point would the width of the structure begin to give you information about the size of the bundle?� To make things simple, lets assume the filaments were in a parallel, flat array with the 8nm filaments spaced by a cross-linking protein like alpha-actinin (about 35nm long).� If your image resolution was 300nm, when would the image of a fluorescent bundle look "wider" than the image of a single filament?� Thanks- Dave

David Knecht, Ph.D.
Professor and Head of Microscopy Facility Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

==============================Original Headers==============================
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Colleagues,

has anybody a recipe or is aware of a kit with which to couple gold to
(strept-)avidin ?

if yes, thanks for a short informal reply!

greetings,

Peter Heimann

--
====================================================

Dr. Peter Heimann Raum: W7-107 / Tel.: +49-(0)521-106-5628
Universitaet Bielefeld Fax: +49-(0)521-106-5654
"Zellbiologie / Cell Biology" W7-107
33501 Bielefeld, Germanywww.uni-bielefeld.de/biologie/cellbio

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Dear Dr. Heimann

From experience I recommend to choose streptavidin over avidin. The
high IEP of avidin makes it hard to obtain cluster free conjugates.
Coupling procedures have been documented in early immunogold conjugate
literature. An important factor is the coupling pH which should be equal
or preferably slightly higher than the IEP of your streptavidin.
Please feel free to contact me off list if you require more
information, we will be happy to help.

Kind regards,
Jan Leunissen
Aurion - ImmunoGold reagents

i: www.aurion.nl

Colleagues,

has anybody a recipe or is aware of a kit with which to couple gold to
(strept-)avidin ?

if yes, thanks for a short informal reply!

greetings,

Peter Heimann

--
====================================================

Dr. Peter Heimann Raum: W7-107 / Tel.: +49-(0)521-106-5628
Universitaet Bielefeld Fax: +49-(0)521-106-5654
"Zellbiologie / Cell Biology" W7-107
33501 Bielefeld, Germany
www.uni-bielefeld.de/biologie/cellbio

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Dear David,
Another consideration is the distribution of the fluorophors along
the filament(s). Variations of number of fluorophors will fold in
statistically with other variations in fluorescence, so a single
filament will have variations along its length and as a function of
time, so it will be more complicated than a linear superposition of
Airy discs. (It is the wave amplitudes that add linearly, and the
resulting intensities depend on the relative phases.) This might be
indistinguishable from a filament of varying size. Of course, a
sufficiently long time average will tend toward a diffraction profile
with a width related ti the resolution.
Yours,
Bill

On May 2, 2013, at 3:49 PM, david.knecht-at-uconn.edu wrote:

}
}
} I am not getting much response, so let me try to explain my
} interpretation of the problem. If you consider a single filament,
} then you expect to image a linear extension of the diffraction
} pattern (airy disk) of each fluorescent point source along the
} filament projected onto the chip. As a diffraction pattern, the
} emission will decrease stochasitically as you move away from the
} filament. In theory, if you wait long enough and acquire enough
} photons over a long time, the diffraction pattern spreads to
} decreasingly represented orders of the pattern. So what you call
} the "edge" of the filament as you image it is arbitrary and
} determined by the sensitivity of your detector and the imaging
} conditions. Now as you increase the number of filaments in a
} bundle, the total fluorescence output from the volume increases. If
} you were to acquire an image with the same gain and integration as
} you did for the single filament, you would see a "wider" structure
} because the higher orders !
} of the pattern now become more readily visible due to the brighter
} signal and more photons coming from the same "sub-resolution"
} volume. If you normalize for the total fluorescence, I would expect
} the single filaments to look just like the bundles up to the point
} where you exceed optical resolution (about 8 filaments when you are
} nearing 300nm wide). Of course a real bundle is much more tightly
} packed as a cylinder, so it would take many more filaments to look
} larger. But if you do not normalize, I think the bundle will look
} larger than a single filament before you exceed the resolution
} limit. My greater point is that the imaging conditions and detector
} will have an effect on what you call the "size" of an object and
} this is a different set of considerations from that we normally
} think about as resolving of multiple objects from each other. Dave
}
} On Apr 29, 2013, at 2:28 PM, david.knecht-at-uconn.edu {mailto:david.knecht-at-uconn.edu
} } wrote:
}
}
}
}
}
}
} } I had a question on an exam I gave recently and now I am trying to
} } figure out what should be a correct answer. The question was
} } whether you could tell an actin filament (8nm) from a bundle of
} } filaments by fluorescence microscopy. They were supposed to think
} } about image resolution and brightness in their answer. Obviously,
} } you would be unlikely to be able to resolve filaments in a bundle
} } since they are so tightly packed. But a bundle should be much
} } brighter than a filament if you had both to compare.
} } My question is, at what point would the width of the structure
} } begin to give you information about the size of the bundle? To
} } make things simple, lets assume the filaments were in a parallel,
} } flat array with the 8nm filaments spaced by a cross-linking protein
} } like alpha-actinin (about 35nm long). If your image resolution was
} } 300nm, when would the image of a fluorescent bundle look "wider"
} } than the image of a single filament? Thanks- Dave
} }
} } David Knecht, Ph.D.
} } Professor and Head of Microscopy Facility
} } Department of Molecular and Cell Biology
} } U-3125
} } 91 N. Eagleville Rd.
} } University of Connecticut
} } Storrs, CT 06269
} } 860-486-2200
} } 860-486-4331 (fax)
}
}

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Email: modla-at-dbi.udel.edu
Name: Shannon Modla

Organization: University of Delaware

Title-Subject: [Filtered] Lowicryl HM20 monostep

Message: Hi Everyone,

I was wondering if anyone has any suggestions on how to re-orient samples embedded in Lowicryl HM20
monostep resin for ultrathin sectioning. In the past, I tried using superglue, but it reacted with
the Lowicryl and made the bottom of the block in contact with the glue gooey. I tried with 2
different tubes of superglue, thinking there was something wrong with the glue, but I had the same
result both times. I also tried a 2 part epoxy with a quick setup time, but it didn't secure the
block as well as superglue usually does with other resins. Has anyone had any similar experience?
Does anyone have any tried-and-true ways that has worked for them to reorient embedded samples in
Lowicryl HM20?

Thanks,
Shannon

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Email: wadowska-at-upei.ca
Name: Dorota Wadowska

Organization: Atlantic Veterinary College at UPEI

Title-Subject: [Filtered] H 7000 clogged cooling lines

Message: Hi all,
Cooling lines in our H 7000 are clogged. We try to flush the system with pressurized water but it
helps little. The debris that comes out is green and hard like maybe copper oxide. Do you now of any
treatment that would remedy his problem without damaging the scope? Here is the link to pictures
depicting debris that comes out of the cooling lines
https://dl.dropboxusercontent.com/u/47838666/image001.jpg
Thanks
Dorota

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Citric Acid is used to clear cooling lines of accumulated malachite, 10g per gallon of water.

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That's some ugly stuff.

I'd try CLR. I've had plugged lines and tried everything from Prestone radiator cleanout, acetic acid mixtures, to floor strippers. Actually the phosphoric acid based floor stripper worked the best, but I can't find it any more. CLR worked the second best.

But first, see if your material dissolves in CLR. If it does, I'd add half a galleon to my recirculation water and leave it over night, maybe over the week-end. If you're using tap water and not recirculating water, you need to make a homemade sump with pump and plumb it in to your microscope.

Good luck!!

Frank

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Email: wadowska-at-upei.ca
Name: Dorota Wadowska

Organization: Atlantic Veterinary College at UPEI

Title-Subject: [Filtered] H 7000 clogged cooling lines

Message: Hi all,
Cooling lines in our H 7000 are clogged. We try to flush the system with pressurized water but it
helps little. The debris that comes out is green and hard like maybe copper oxide. Do you now of any
treatment that would remedy his problem without damaging the scope? Here is the link to pictures
depicting debris that comes out of the cooling lines
https://dl.dropboxusercontent.com/u/47838666/image001.jpg
Thanks
Dorota

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Dorota,

I've used both CLR cleaner (this is the brand name; for bathrooms,
mostly) and hydrogen peroxide to the clean the chiller lines on our TEM
- by the service engineer's recommendation.
One bottle of CLR in 5 gallons of chiller water, let run for ~4 hours or
overnight, then drain the tank and flush repeatedly until only clear
water comes out.
Or 1 liter 3% H2O2 or 100 mL 30% H2O2 and do the same. This worked when
the CLR didn't.

Works a treat.

Phil

} Email: wadowska-at-upei.ca
} Name: Dorota Wadowska
}
} Organization: Atlantic Veterinary College at UPEI
}
} Title-Subject: [Filtered] H 7000 clogged cooling lines
}
} Message: Hi all,
} Cooling lines in our H 7000 are clogged. We try to flush the system with pressurized water but it
} helps little. The debris that comes out is green and hard like maybe copper oxide. Do you now of any
} treatment that would remedy his problem without damaging the scope? Here is the link to pictures
} depicting debris that comes out of the cooling lines
} https://dl.dropboxusercontent.com/u/47838666/image001.jpg
} Thanks
} Dorota
}
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Like so much in science, the best way is the most tedious and time-consuming. I polymerize my tissues in flat bottom BEEM capsules, cut out the tissue as a medium size chunk using a Dremel moto-tool and re-embed in fresh Lowicryl in a new BEEM capsule. If you cut an appropriate size piece of plastic after the first embedding, it will stand up straight in the second BEEM capsule and retain its orientation during the second polymerization. I always feel like a diamond cutter when I am trimming the first block to get the shape and orientation for re-embedding. Good luck. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

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Email: modla-at-dbi.udel.edu
Name: Shannon Modla

Organization: University of Delaware

Title-Subject: [Filtered] Lowicryl HM20 monostep

Message: Hi Everyone,

I was wondering if anyone has any suggestions on how to re-orient samples embedded in Lowicryl HM20 monostep resin for ultrathin sectioning. In the past, I tried using superglue, but it reacted with the Lowicryl and made the bottom of the block in contact with the glue gooey. I tried with 2 different tubes of superglue, thinking there was something wrong with the glue, but I had the same result both times. I also tried a 2 part epoxy with a quick setup time, but it didn't secure the block as well as superglue usually does with other resins. Has anyone had any similar experience?
Does anyone have any tried-and-true ways that has worked for them to reorient embedded samples in Lowicryl HM20?

Thanks,
Shannon

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I can't provide any guarantees in regard to potential possibility of
"damaging the scope", therefore "use at your own risk, yada, yada,
yada", but over past two years I've applied following technique to two
instruments, one is to clean clogged cooling line in objective lens
(FEI) another is to remove deposits from cooling lines and chiller tank
(Hitachi), and so far - so good.

1. Pour liberal amount of "Micro-90" (available from both Cole-Parmer
and Sigma-Aldrich) into the tank of recirculating chiller. I've used
about 200CC for a small Neslab chiller, about 0.5L for dual-loop
Haskris. Let it run overnight - by the morning water will be green and
all (or nearly all) of deposits from the tubing and walls of the tank
will be dissolved.

2. Drain (now green) water from the cooling system, purge with
compressed air, refill with DI, and run chiller for a 15min. or so. If
visible deposits come out during air purge or accumulate in the tank
after the refill, then repeat Micro-90 treatment.

3. Drain cooling system, purge with compressed air, and flush with
regular and then DI water. I've done at least 3 full refills with tap
water, followed by at least two refills with DI, in each case. Fill
with DI and run at least overnight, or for a day or two.

4. Drain system, purge with compressed air, and refill with DI one last
time - maybe overkill, but I did not want to have any traces of Micro-90
remain in the system.

One of the tools treated in this way is running over a year without
visible signs of corrosion in the cooling system (no green particles
coming out, or accumulating in the chiller tank), another one was
treated just a couple of months ago.

Again - I make no guarantees for the safety of such treatment, realize
that absence of visible signs of corrosion does not mean that nothing is
happening inside of the cooling lines, etc... so use (or don't) at your
own risk :-)

Good luck!

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

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} Email: wadowska-at-upei.ca
} Name: Dorota Wadowska
}
} Organization: Atlantic Veterinary College at UPEI
}
} Title-Subject: [Filtered] H 7000 clogged cooling lines
}
} Message: Hi all,
} Cooling lines in our H 7000 are clogged. We try to flush the system with pressurized water but it
} helps little. The debris that comes out is green and hard like maybe copper oxide. Do you now of any
} treatment that would remedy his problem without damaging the scope? Here is the link to pictures
} depicting debris that comes out of the cooling lines
} https://dl.dropboxusercontent.com/u/47838666/image001.jpg
} Thanks
} Dorota
}
} Login Host: 137.149.102.148
} Listserver Email Form V - 20120416
} ---------------------------------------------------------------------------
}

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Contents Retrieved from Microscopy Listserver Archives
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Dear all,

if you have (pre-)registered for this meeting already, please register
again at http://nanocrystallography.net/ASCI2013 in order to receive
an automatic confirmation email that you may need for your travel
arrangements / tax documentation / future reimbursements. This
personalized email will be the only "receipt" that we are going to
issue.

If you are pondering attending this meeting, please check out
http://nanocrystallography.net/ASCI2013 for full details.

The free two day workshop, "Advances in Structural and Chemical
Imaging", by the PREMIER* network on May 29 and 30, 2013, at CAMCOR,
University of Oregon, Eugene, OR (* Pooled Resources for Electron
Microscopy Informatics Education and Research). There are 22 invited
speakers and up to 25 posters. Space is limited and time flies, so
please register soon.

Scopes

1. structural analysis of organics, membrane proteins and macromolecules
2. structural analysis of inorganics and nanocrystals
3. structural analysis of crystalline surfaces and interfaces, 2D
crystallography, surface reconstructions
4. full chemical analysis of crystalline phases by combining
atomic level structural and spectroscopic analyses
5. polycrystal orientation and phase mapping/analysis, texture
and interface analyses at the nanometer length scale

6. large scale microscope data acquisition and processing
7. reconstruction algorithms for 3-D imaging and compressed sensing

We envision an interdisciplinary meeting that bridges the major divide
in this field between the structural biology and material science
communities. The multitude of methods being employed are linked by
common themes in their desire to achieve chemical specificity, high
spatial resolution (atomic in some cases) and the need to acquire,
process and analyze large data sets to identify signature events.

Attached are the schedule (22 invited speakers), driving directions
and maps, as well as a comprehensive event flyer.

Required registration at http://nanocrystallography.net/ASCI2013.

You are invited to present posters, additional registrations for
poster presenter (at the above website) will be necessary in these
cases.

Vendors who would like to represent their companies are to register in
addition with Randy and Joe (smithran-at-pdx.edu,
lymphocyte58-at-hotmail.com, joegray3-at-hotmail.com) of the Pacific
Northwest Section of the Microscopy Society of America.

The scopes of the meeting involve applications of the following methods:

- high-resolution transmission electron microscopy (TEM)
- scanning transmission electron microscopy (STEM)
- analytical electron microscopy (TEM): electron energy loss
spectroscopy (EELS), energy dispersive X-ray spectroscopy (EDXS), and
other spectroscopies
- precession electron diffraction (PED)
- scanning nano-beam transmission electron diffraction (SNBD)
- convergent beam electron diffraction (CBED)
- direct space tomography with STEM and TEM
- transmission electron diffraction tomography (DT)
- backscattered electron diffraction (EBSD) in combination
with EDXS in scanning electron microscopy (SEM)
- advanced electronic data management, telemicroscopy, open
access databases
- low energy electron diffraction (LEED)
- reflection high-energy electron diffraction (RHEED)
- scanning tunneling microscopy (STM) and all kinds of
scanning probe microscopies (SPM)
- X-ray microscopy, tomography and spectroscopy
- advanced light microscopy
- mass spectroscopy
- magnetic resonance imaging, nuclear magnetic resonance spectroscopy

--
Peter Moeck, PhD (Dr. rer. nat. Crystallography)
Associate Professor of Physics

Nano-Crystallography Group:
http://nanocrystallography.research.pdx.edu/nano-crystallography-group.
Free (as in free speech rather than free beer) crystallographic
information and interactive 3D visualizations of crystal structures
and morphologies with educational relevance:
http://nanocrystallography.research.pdx.edu/databases.

Portland State University
Department of Physics
P.O. Box 751, Portland, OR 97207-0751
Tel.: 503 725 4227, Departmental Fax.: 503 725 2815
pmoeck-at-pdx.edu
www.physics.pdx.edu/~pmoeck

Office 404, Science Research and Teaching Center
1719 SW 10th Ave, Portland, OR 97201

Office hours during quarters Tuesdays and Thursdays 12:00 to 12:30 and
per appointment

North American mirror of the Crystallography Open Database, COD,
http://nanocrystallography.org, with more than 210,000 carefully
evaluated datasets for download and interactive visualization in 3D,
for more interactive 3D display options try:
http://nanocrystallography.research.pdx.edu/search/codmirror/
Webportal: Open Access Crystallography Resource Portal
http://nanocrystallography.net.

"... no hell below us, above us only sky ..." John Lennon
"... der wackere Schwabe forcht sich nit ..." Albert Einstein
(inspired by Ludwig Uhland's poem)
"Was auch immer geschieht: Nie duerft ihr so tief sinken, von dem
Kakao, durch den man euch zieht, auch noch zu trinken." Erich Kaestner
"... the only thing needed for a happy life is a project one is
enthusiastic about." John C. H. Spence, Acta Cryst. A 69 (2013) 25-33

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Hi Dorota and Valeriy,
One possible problem is if the corrosion in the cooling lines, which
caused the clogging, has proceeded to the point that the tubing in the
lenses is leaky or weak enough that removing the deposits will allow
leaks to develop. When you clean the lines, monitor for these
possible leaks. If everything is OK, maintain the cooling water at a
slightly basic pH (7.5 to 8.0) to prevent further corrosion, and, if
warranty or service contract permit, add a corrosion inhibitor to the
water. When I was in NY State, I used a product called AquaTreet 42
from Aqua Labs, and in California I used an equivalent Skasol
product. Check the pH and inhibitor levels--the products come with
test kits--every month and adjust levels appropriately. Check with
the manufacturer or service person to be sure that these additions
will not cause any problems with either the scope or your contract,
but they have worked on all the scopes I've been in charge of.
Yours,
Bill

On May 6, 2013, at 10:08 AM, vray-at-partbeamsystech.com wrote:

}
} I can't provide any guarantees in regard to potential possibility of
} "damaging the scope", therefore "use at your own risk, yada, yada,
} yada", but over past two years I've applied following technique to two
} instruments, one is to clean clogged cooling line in objective lens
} (FEI) another is to remove deposits from cooling lines and chiller
} tank
} (Hitachi), and so far - so good.
}
} 1. Pour liberal amount of "Micro-90" (available from both Cole-Parmer
} and Sigma-Aldrich) into the tank of recirculating chiller. I've used
} about 200CC for a small Neslab chiller, about 0.5L for dual-loop
} Haskris. Let it run overnight - by the morning water will be green and
} all (or nearly all) of deposits from the tubing and walls of the tank
} will be dissolved.
}
} 2. Drain (now green) water from the cooling system, purge with
} compressed air, refill with DI, and run chiller for a 15min. or so. If
} visible deposits come out during air purge or accumulate in the tank
} after the refill, then repeat Micro-90 treatment.
}
} 3. Drain cooling system, purge with compressed air, and flush with
} regular and then DI water. I've done at least 3 full refills with tap
} water, followed by at least two refills with DI, in each case. Fill
} with DI and run at least overnight, or for a day or two.
}
} 4. Drain system, purge with compressed air, and refill with DI one
} last
} time - maybe overkill, but I did not want to have any traces of
} Micro-90
} remain in the system.
}
} One of the tools treated in this way is running over a year without
} visible signs of corrosion in the cooling system (no green particles
} coming out, or accumulating in the chiller tank), another one was
} treated just a couple of months ago.
}
} Again - I make no guarantees for the safety of such treatment, realize
} that absence of visible signs of corrosion does not mean that
} nothing is
} happening inside of the cooling lines, etc... so use (or don't) at
} your
} own risk :-)
}
} Good luck!
}
}
} Valery Ray
}
} On 5/6/2013 9:18 AM, microscopylistserver-noreply-at-microscopy.com
} wrote:
} }
} } Email: wadowska-at-upei.ca
} } Name: Dorota Wadowska
} }
} } Organization: Atlantic Veterinary College at UPEI
} }
} } Title-Subject: [Filtered] H 7000 clogged cooling lines
} }
} } Message: Hi all,
} } Cooling lines in our H 7000 are clogged. We try to flush the system
} } with pressurized water but it
} } helps little. The debris that comes out is green and hard like
} } maybe copper oxide. Do you now of any
} } treatment that would remedy his problem without damaging the scope?
} } Here is the link to pictures
} } depicting debris that comes out of the cooling lines
} } https://dl.dropboxusercontent.com/u/47838666/image001.jpg
} } Thanks
} } Dorota

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Hello All,
Our UV lamp burnt out after 10 years, not certain how many hours of use, and trying to get a replacement through Leica is extremely expensive. Has anyone replaced their "CATHODEON 1W 6W MADE IN ENGLAND" lamp recently? The EM UV fits atop Leica's cryo preparation unit or the AFS first generation unit.
John

==============================Original Headers==============================
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Hi Everyone,

We still have a few spots left in the 3D CLEM course! Get in touch with Garnet if you're interested!

Preliminary programme for the inaugural UBC 3D CLEM course, June 14-17, 2013

This course is designed for life science researchers/microscopists who are looking to learn 3D Correlative Microscopy. Our instructors will guide you with a series of lectures and practical demonstrations. We will introduce a sample (or two) at the beginning of the course and take that one sample through microCT, confocal/2P, dual beam FESEM and TEM tomography and into advanced 3D image processing. Sample preparation will include both chemical fixation as well as high pressure freezing.

Synopsis:

When: June 14-17, 2013
Where: UBC Bioimaging Facility and UBC Centre for High-Throughput Phenogenomics, University of British Columbia, Vancouver, BC. Canada
Support: Systems for Research (SFR), FEI Company, Edge Scientific, Leica Microsystems, Olympus Canada, TILL Photonics and VSG
Who: 10-12 students, preferably life sciences PhD students, Post docs, microscopy facility managers/directors/technicians.
Course fee: $2500 CDN (subject to change) and will include 5 nights accommodation, breakfast, lunch and one dinner.

For more information contact Garnet Martens at garnet.martens-at-botany.ubc.ca.

Preexisting knowledge of both light and electron microscopy necessary.

Lectures will include:

Principles of CLEM
CLEM methods
microCT for CLEM
Dual beam SEM for CLEM
Tomography
CryoEM sample preparation
Confocal/2P for CLEM
Image processing and analysis
Practical sessions:
microCT scanning
confocal/2p imaging
sample handling for both chemical fixation and high pressure freezing
FIB/SEM
Tomography
Image processing and analysis

Instructors:

Elizabeth Fischer, Rocky Mountain Labs, Hamilton, MT
Vinod Nair, Rocky Mountain Labs, Hamilton, MT
Bill Rice, New York Structural Biology Center
Kim Rensing, Leica Microsystems
Richard Gursky, FEI Company
Cliff Matheson, FEI Company
Christian Wietholt, VSG
TILL Photonics
Farid Jalali, Olympus Canada
Nancy Ford, UBC Centre for High-Throughput Phenogenomics
Gethin Owen, UBC Centre for High-Throughput Phenogenomics
Kevin Hodgson, UBC Bioimaging Facility
Derrick Horne, UBC Bioimaging Facility
Brad Ross, UBC Bioimaging Facility
Garnet Martens, UBC Bioimaging Facility

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13, 35 -- Subject: 2nd Announcement: Inaugural UBC 3D CLEM course, June 14-17, 2013
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****COMMERCIAL POSTING****

Microscopy Sales Representative

We are looking for a motivated microscopy sales professional that is looking to be a part of a growing company. JH Technologies has been in business since 1987 providing microscopy research instruments and industrial equipment in Northern California and are looking to expand our sales talent to support our business. Our business is focused on providing complete optical and digital imaging solutions to our client base.

In this position you will be selling life science research instruments from Leica Microsystems as well as additional complementary products targeting life science research institutions, biotech, biomedical, and electronics markets. Our target is to support, develop, and grow our business with major accounts, and additionally to prospect and find new opportunities within our markets.

The successful candidate will grow sales revenue by maintaining positive relationships with current customers, cultivating new customers and market opportunities, creating compelling proposals, demonstrating and installing equipment, and always providing first class support. The candidate will also be required to maintain and regularly update our CRM system, create weekly call plans and customer contact reports, prepare annual territory plans, participate in local and national trade shows, seminars, and trainings.

Occasional travel to participate in seminars, trainings, product launches, and trade shows should be expected.

Requirements

* Scientific equipment sales experience with a proven track record of growth and exceeding sales quotas; formal professional sales training (consultative selling style)

* Knowledge of the local customer base with established contacts: end users, management/decision makers, and commercial personnel (buyers, purchasing agents)

* Experience with microscopes, imaging systems, optical measuring systems, or other applicable skills.

* Minimum Bachelor's degree in biology or applicable science discipline or scientific field or 5 years significant applications experience in the life science market place.

* Strong organizational skills, along with a track record of responsive, timely and effective territory management.

* Strong competitive drive - must want to win, grow and excel

* Excellent communication skills both verbal and written; strong presentation skills needed

* Working knowledge of MS Office and Salesforce.com is a must.

* Ethical and honest approach to business success

* Must be goal oriented and focused on achieving challenging sales growth targets

* Must live in the territory and have reliable transportation

Cultural Values

* Professionalism

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* Team Player

* Personal success matched with Company success

* Open communication/2 way communication

* Bottom line oriented

* Balanced between business and family

* Willingness to learn

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Additional values and traits

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* Technical confidence

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* Have used a CRM

Please send CV's to mharrel-at-jhtechnologies.com
Please do not call.

Regards,

Micah Harrel
Vice President
JH TECHNOLOGIES, INC.
225 Hammond Avenue
Fremont, CA 94539
cell 408.640.0774
tel 408.436.6336 x303
fax 408.436.6343
mharrel-at-jhtechnologies.com

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Email: hwitkiewicz-at-ucsd.edu
Name: Halina Witkiewicz

Organization: PRISM

Title-Subject: [Filtered] Ua/Pb stain process unknown debris

Message: You could skip the Ua/Pb staining if you use digital camera to capture the images. The
digital camera is more sensitive than human eye and one can increase the contrast electronically at
the time of capturing images.
See: http://f1000research.com/articles/2-9/v2

Email: ARD3A-at-virginia.edu
Name: Art Dewey

Organization: Univ. of Va Health Center /Pathology

Title-Subject: [Filtered] Ua/Pb stain process unknown debris

Message: Have suddenly beem plagued with debris after Ua/Pb staining of EMbed-812 embedded thin
sections. I am using the same proceedure I have used for years with no proplem. I have not seen
this type of debris except for one short period of time 3 years ago, never identified and clered up
after a few staining sessions. This time I have had the oroblem for weeks. I have reviewed all my
techniques for changes and replaced all reagents. Problem has no common demoninator. Debris is 1-2
um to 20 um in size, light grey to dark grey, semi-circular to half circle/wedge shape, trans-lucent
to the beam, reactive in the beam. Has anyone seem this and know what it is? Thx

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Email: bob.price-at-uscmed.sc.edu
Name: Bob Price

Organization: Univ South Carolina

Title-Subject: [Filtered] Confocal Microscopy Workshop

Message:

The South Carolina EPSCoR/IDeA Program and the USC School of Medicine Instrumentation Resource
Facility are pleased to announce the 9th annual workshop on Basic Confocal Microscopy. The Workshop
will be held June 10-14 at the USC School of Medicine in Columbia, SC.

Workshop material is directed towards beginning and intermediate users of confocal microscopes and
involves a series of lectures (specimen preparation, labeling strategies, proper set-up of
instrument operating parameters, proper handling of 2D and 3D confocal images in programs such as
Adobe Photoshop and AMIRA), hands on specimen preparation, and time on a number of different point
scanning and spinning disk confocal microscopes.

Companies that have provided equipment and applications experts for past workshops include Andor
Technology, Leica, Nikon, Olympus, Perkin Elmer, Zeiss, Intelligent Imaging Innovations (3i) and
Photometrix. Participants will have ample time for hands on use of the instruments during the workshop

Faculty will include Dr. Jay Jerome of Vanderbilt University, Dr. Ralph Albrecht of the University
of Wisconsin Madison, Dr. John Mackenzie of North Carolina State University, Dr. Tom Trusk of the
Medical University of South Carolina, and Dr. Bob Price of the University of South Carolina.

For further information please see: http://irf.med.sc.edu/ or contact Anna McNeal
(anna.mcneal-at-uscmed.sc.edu {mailto:anna.mcneal-at-uscmed.sc.edu%3cmailto:anna.mcneal-at-uscmed.sc.edu} ) or
Bob Price (bob.price-at-uscmed.sc.edu {mailto:bob.price-at-uscmed.sc.edu%3cmailto:bob.price-at-uscmed.sc.edu} )

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Title-Subject: [Filtered] Hummer II electric diagram

Message: Hello,
Is there anybody who has an electric diagram of Hummer II system?
Thanks,

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Email: ravi.thakkar369-at-gmail.com
Name: Ravi Thakkar

Organization: NIMHANS

Title-Subject: [Filtered] FTIR Imaging of human tissue section Histochemical stained

Message: Dear Listener,
We need to analyse biological sample with FTIR imaging..

Objective of Analysis :
We want qualitative and quantitative analysis of inclusions ,distinguished by staining, So it will
be easy to target FTIR beam.

About sample :
Human tissue section (thickness range 5-12 um )embedded in parafilm wax and collected on glass
slide. Sections are stained with MGT (histochemical stain) for light optical microscope observation.

Queries :
1. Will staining interfere with FTIR analysis. ?
2. Glass slide and parafilm wax will works under IR beam.?

Kindly give your suggestions.

With Thanks & Regards.

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Hi Ravi

The short answer is possibly yes. It depends what you believe the inclusions to be.
Keep in mind that MGT contains Phosphotungstic acid, Aluminium etc. So if you were looking at lung from occupational respiratory exposure there might be an issue. You would have to demonstrate levels higher than the background staining.
We have not used FTIR (only EDS) but we use consecutive sections. The first to stain and localise the area and then mark that region on the second. Sometime the inclusions can be localised in the unstained section with polarised light microscopy.

Hope this helps
Naomi

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Email: ravi.thakkar369-at-gmail.com
Name: Ravi Thakkar

Organization: NIMHANS

Title-Subject: [Filtered] FTIR Imaging of human tissue section Histochemical stained

Message: Dear Listener,
We need to analyse biological sample with FTIR imaging..

Objective of Analysis :
We want qualitative and quantitative analysis of inclusions ,distinguished by staining, So it will
be easy to target FTIR beam.

About sample :
Human tissue section (thickness range 5-12 um )embedded in parafilm wax and collected on glass
slide. Sections are stained with MGT (histochemical stain) for light optical microscope observation.

Queries :
1. Will staining interfere with FTIR analysis. ?
2. Glass slide and parafilm wax will works under IR beam.?

Kindly give your suggestions.

With Thanks & Regards.

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Hi Ravi,

For FTIR, you will need to put your tissue on an IR-reflective slide or an
IR-transmissive slide, but in either case, not glass. IR-transmitting
slides are generally some kind of salt, and all of these are available
from specialist suppliers.

Staining will give an IR signal, certainly.

Why are you doing FTIR analysis rather than any other type of analysis?

regards,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Email: Georgianne.Ciraolo-at-cchmc.org
Name: Georgianne Ciraolo

Organization: Cincinnati Children's Hospital Medical Center

Title-Subject: [Filtered] Negative staining of HIV virus particles.

Message: I recently received a virus sample that had been concentrated by another lab and brought
to me in 100mM ammonium acetate. When I stained the grids there seems to be a precipitate all over
the sample. I was wondering if this could be from the ammonium acetate. Most times I receive my
samples in PBS or something of that sort.

Any opinions would be appreciated.

Thanks

Georgianne Ciraolo
Department of Pathology
Cincinnati Children's Hospital
Cinicinnati, Ohio

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} to me in 100mM ammonium acetate. When I stained the grids there seems to be
} a precipitate all over
} the sample. I was wondering if this could be from the ammonium acetate.
} Most times I receive my samples in PBS or something of that sort.

PBS is the buffer for which I would expect a precipitate, when staining with UAc; but not with Am-Ac buffer. Strange.
The only thing that we always do, is a brief (1 to 2 sec, once) wash on bidistilled water, before the sample is stained with UAc (after staining: only blotting, but no washing). Did you do this, did you try this? I prefer bidistilled (!) water, although many people use 'millipore' water - IMO, this is inferior to bidistilled water, for EM.
Another point: do you have a solution of 1% Phosphotungstic Acid, buffered to about pH 7.0 +/- 0.5 (NaOH)? staining and 'precipitation' is different, usually less precipitates = better, so worth trying.
we always use carbon-coated 400 mesh Cu grids, glow-discharged (if this is of interest).
kind regards,
Reinhard

--
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

next microscopy conferences:
http://www.mc2013.de/
MC2013 in Regensburg, Germany
http://www.imc2014.com/
18th IMC 2014 in Prague, Czech Rep.

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Ammonium acetate is volatile at low vacuum pressures. Researchers often use dialysis to exchange salts like NaCl with ammonium acetate so they can later evaporate off it off downstream. I use to do this with a lyophilizer but you could also use a rotary evaporator (also called a rotavap). A rotavap is small centrifuge that pulls a vacuum on the sample as it is being spun. Then re-suspend your sample in deionized water or PBS. Tom Phillips

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

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Organization: Cincinnati Children's Hospital Medical Center

Title-Subject: [Filtered] Negative staining of HIV virus particles.

Message: I recently received a virus sample that had been concentrated by another lab and brought to me in 100mM ammonium acetate. When I stained the grids there seems to be a precipitate all over the sample. I was wondering if this could be from the ammonium acetate. Most times I receive my samples in PBS or something of that sort.

Any opinions would be appreciated.

Thanks

Georgianne Ciraolo
Department of Pathology
Cincinnati Children's Hospital
Cinicinnati, Ohio

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The NEW YORK SOCIETY OF EXPERIMENTAL MICROSCOPISTS will hold their Spring meeting on Thursday May 16, 2013 at Rockefeller University. Dr. John Condeelis is the invited speaker. Please see the announcement below.

NYSEM Spring Meeting
Thursday May 16, 2013, 5:30 PM

Rockefeller University
1230 York Ave, New York, NY
Enter campus at York Avenue & 66th street
Carson Family Auditorium, Greenberg Building (CRC)

John S. Condeelis, Ph.D.
Professor & Co-Chair of Anatomy & Structural Biology
Albert Einstein College of Medicine

"High Resolution Single Cell Imaging in Mammary Tumors"

Multi-photon microscopy allows the observation of tumor cell behavior and cell fate at single cell resolution in vivo in deep tissue. Imaging in deep tissue locations in live animals in real time has revealed molecular mechanisms associated with complex cancer cell behavior leading directly to new markers of metastatic risk and treatment response.

MEETINGS ARE FREE AND OPEN TO ALL!

Frank Macaluso
Senior Associate in Anatomy and Structural Biology
Administrative Director and
Director of Electron Microscopy
Analytical Imaging Facility
Albert Einstein College of Medicine
1300 Morris Park Avenue
Bronx, NY 10461

(718) 430-3547
frank.macaluso-at-einstein.yu.edu
http://www.einstein.yu.edu/aif

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Hi All,

We need to prepare styrofoam for imaging cell structure and thickness of
cell walls. Most desirable would be to use SEM so that sample size would
be large enough to get statistically accurate data. Initial thought was to
embed and then smooth off the surface
so that SEM could be used. However, most embedding reagents will damage
the styrofoam and we are also concerned about the chemicals causing
thickness changes within the sample.

It might be possible to cut a smooth surface with a very sharp scalpel or
razor blade. Then very low kV SEM might be a possibility as there is
concern that higher kV might damage the sample.

We would appreciate feedback from anyone who has experience working with
styrofoam.

Thanks,
Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

==============================Original Headers==============================
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10, 30 -- From: "Sherman, Debra" {dsherman-at-purdue.edu}
10, 30 -- To: "message to: MSA list" {microscopy-at-microscopy.com}
10, 30 -- Subject: imaging styrofoam
10, 30 -- Thread-Topic: imaging styrofoam
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Hi All,

We are looking for information from users of SDD EDS systems on very high
end FEG-TEMs (300kV). We would appreciate your assessment of whatever
system you are using as to sensitivity, count rates (especially for
mapping), ease of software use by novice
users, and ability to upgrade after the install as needed. Are there
add-on programs to increase functionality that you would recommend for a
multi-user facility?

Also how was your customer experience with the specific vendor during
initial purchase and install, as well as vendor support for trouble
shooting after install? Are you totally satisfied with your choice and, in
hind sight, would you chose differently
if purchasing again?

No vendors please. We really want to hear from the end users.

Thanks,
Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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10, 30 -- Subject: EDS for high voltage TEM
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Hi Debby,
I had this problem a year before. The only fast way to image styrofoam is to cut it with a NEW cooled razor-blade under liquid
nitrogen. Then you will get this quality of the images:
http://www.electronmicroscopy.info/shop_materials.htm, starting from image 27 (polystyrene)
Image 31 on this page shows what you are looking for. Imaged mostly with backscatterd detectors at 20 kV, LaB6, coating is
Platinium, coating-time ca. 150% of normal coating-time used...

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 11.05.13 20:16, schrieb dsherman-at-purdue.edu:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi All,
}
} We need to prepare styrofoam for imaging cell structure and thickness of
} cell walls. Most desirable would be to use SEM so that sample size would
} be large enough to get statistically accurate data. Initial thought was to
} embed and then smooth off the surface
} so that SEM could be used. However, most embedding reagents will damage
} the styrofoam and we are also concerned about the chemicals causing
} thickness changes within the sample.
}
} It might be possible to cut a smooth surface with a very sharp scalpel or
} razor blade. Then very low kV SEM might be a possibility as there is
} concern that higher kV might damage the sample.
}
} We would appreciate feedback from anyone who has experience working with
} styrofoam.
}
} Thanks,
} Debby
}
}
} Debra Sherman, Founder & CSO
} DS imaging LLC
} Purdue Technology Center
} 1281 Win Hentschel Blvd
} West Lafayette, IN 47906
} E-mail: debby.sherman-at-dsimagingllc.com
} www.dsimagingllc.com
} Mobile: 765-418-8540
}
}
}
}
} ==============================Original Headers==============================
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} 10, 30 -- From: "Sherman, Debra" {dsherman-at-purdue.edu}
} 10, 30 -- To: "message to: MSA list" {microscopy-at-microscopy.com}
} 10, 30 -- Subject: imaging styrofoam
} 10, 30 -- Thread-Topic: imaging styrofoam
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} 10, 30 -- Date: Sat, 11 May 2013 18:07:58 +0000
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Hi Debby,

the foam sample preparation protocols including from Stefan (Thank
you!!) are being collected here:
http://confocal-manawatu.pbworks.com/w/page/36915707/Foam%20Sample%20Preparation

Probably, methods developed for the krispies samples could be useful too:
http://confocal-manawatu.pbworks.com/w/page/16346940/Krispies-Sample-Preparation
In the case of imaging of the large area the nm-scale resolution is
probably not required. Light microscopy would give you sub-micron
resolution and could probably be sufficient for your project.

Cheers,
Dmitry

Advanced Knowledge Management
for MICROSCOPY and Image Analysis
________________________________
Dmitry Sokolov, Ph.D.
Mob: +64 21 063 5382
dmitry.v.sokolov-at-gmail.com

-------- Original Message --------

Hi Debby,
I had this problem a year before. The only fast way to image styrofoam is to cut it with a NEW cooled razor-blade under liquid
nitrogen. Then you will get this quality of the images:
http://www.electronmicroscopy.info/shop_materials.htm, starting from image 27 (polystyrene)
Image 31 on this page shows what you are looking for. Imaged mostly with backscatterd detectors at 20 kV, LaB6, coating is
Platinium, coating-time ca. 150% of normal coating-time used...

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt:http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 11.05.13 20:16, schriebdsherman-at-purdue.edu:
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} Hi All,
}
} We need to prepare styrofoam for imaging cell structure and thickness of
} cell walls. Most desirable would be to use SEM so that sample size would
} be large enough to get statistically accurate data. Initial thought was to
} embed and then smooth off the surface
} so that SEM could be used. However, most embedding reagents will damage
} the styrofoam and we are also concerned about the chemicals causing
} thickness changes within the sample.
}
} It might be possible to cut a smooth surface with a very sharp scalpel or
} razor blade. Then very low kV SEM might be a possibility as there is
} concern that higher kV might damage the sample.
}
} We would appreciate feedback from anyone who has experience working with
} styrofoam.
}
} Thanks,
} Debby
}
}
} Debra Sherman, Founder & CSO
} DS imaging LLC
} Purdue Technology Center
} 1281 Win Hentschel Blvd
} West Lafayette, IN 47906
} E-mail:debby.sherman-at-dsimagingllc.com
} www.dsimagingllc.com
} Mobile: 765-418-8540
}
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Organization: San Francisco Microscopical Soc iety

Title-Subject: [Filtered] Brian Ford Talk on Early Microscopy, May 15, Oakland, CA

Message: Poster: http://www.scribd.com/doc/140119964/Brian-Ford-Announcement

We are privileged to present renowned British scientist Brian J. Ford, Wednesday May 15 at Merritt
College, 12500 Campus Drive, Oakland, in the Student Lounge, Building R, room 110. Talk begins at
7:30 PM. Doors open at 6:30. Event is free and open to the general public. Refreshments will be
served. Parking is on the south side of the campus.

If you have had the pleasure and privilege of hearing Brian Ford speak, you know that he always
surprises his audience with his insightful presentations on science, and microscopy in particular.

We have invited Brian to speak about the astonishing views that early microscopes presented to
scientists of the past. He takes us back to visit the Netherlands in the 1600s and also to London,
where Leeuwenhoek and Hooke gave birth to the science of microscopy. Professor Ford has recently
succeeded for the first time in history in obtaining videos using digital
image-capture that recreates precisely what was seen by microscopists in the 1600s.

His presentation of this research to the Royal Society of London was a memorable occasion, and today
he shows further images from the dawn of microscopy in this vivid presentation.

The scientific journal Nature reported: "Ford is the world's leading expert on the topic, and what
he has to say here makes a good deal of sense."

Brian Ford's presentation is proudly sponsored by the following organizations and businesses:
Merritt College Microscopy Program, San Francisco Microscopical Society, Northern California Society
for Microscopy, Forensic Analytical Laboratories, American Society of Trace Evidence Examiners,
California Association of Criminalists, Merritt College Microscopy Club, and Merritt College Biology
Club.

We will be holding a pre-Meeting dinner for Brian Ford before his talk on May 15th, meeting at 5:00
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The 2013 Microscopy and Microanalysis meetings in Indianapolis, IN are
less than 3 months away and we could use your help to make sure things
run smoothly. Student bursaries or volunteers are needed to do such
things as provide support in the different symposia, staff the volunteer
office, monitor use of the Internet Caf�, and help with vendor tutorial
sign-up.

Student bursaries will earn $10/hour for assisting with any of the
tasks mentioned above (paid by check at the end of the meetings). There
is an added bonus of $10 cash for each morning and/or afternoon session
worked to help with meals. Although not paid per hour, volunteers are
eligible for the $10 cash towards meals and both groups will get a
meeting shirt.

We can*t do it without help from you, so please consider serving as a
student bursary or volunteer. If anyone has any questions about the
bursary/volunteer program, or would like to participate, please contact
me.

Don't forget early registration deadline is June 24!

Amanda Lawrence
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Mississippi State University
662-325-3019
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Email: rwysolmerski-at-hsc.wvu.edu
Name: Robert Wysolmerski

Organization: West Virginia University

Title-Subject: [Filtered] LM and NLO Core Facility Manager

Message: Center for Neuroscience at West Virginia University School of Medicine, is seeking to hire
a Core Facility Manager (Biomedical Engineer) to manage a light microscopic imaging and image
analysis facility that includes non-linear optical microscopy. The Biomedical engineer will manage
a light microscopic imaging and image analysis facility that includes non-linear optical microscopy
capabilities and assist facility users (faculty, postdoctoral fellows, students and staff) at West
Virginia University. Responsibilities are functional in nature, and performed under limited
supervision. Specific tasks include, but are not limited to: 1) Assist faculty, postdoctoral
fellows, students and staff at West Virginia University, 2) Monitor equipment, setup and align
lasers, 3) Search for updates and find new ways of applying it to the research being conducted at
the Sensory Neuroscience Research Center, 4) Implement new technologies as they arise, 5) Implement
and optimize experimental protocols for fluorescence imaging procedures, such as FRET and
ratiometric imaging, intrinsic imaging techniques, programming development for image analysis, and
identification of new equipment for acquisition, 6) Develop methods and software for new
experimental procedures, 7) Manage computers associated with imaging equipment, 8) Assist
investigators in troubleshooting hardware problems 9) Train researchers in the use of image analysis
software, 10) Help the Core Director advertise the core facility to the university community.
Participate in biotechnology workshops organized at WVU to educate the scientific community, 11)
Identify new equipment for acquisition and advise the Core Director, 12) Participate in the
submission of grant applications to acquire additional equipment for the imaging core facility and
13) Identification of new equipment for acquisition and implementation of new technologies for
enhancement of the core facility.
For additional information contact Robert Wysolmerski at: rwysolmerski-at-hsc.wvu.edu

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Email: michael.cammer-at-med.nyu.edu
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Title-Subject: [Filtered] Leica and Nikon threads

Message: Do you know if the Nikon and Leica objectives us a compatible threading?
Thank you!

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Email: sachin.kumar-at-cchmc.org
Name: Sachin Kumar

Organization: Post Doc/ CCHMC, Cincinnati

Title-Subject: [Filtered] Neutrophil transendothelial migration

Message: Hi,

I am trying to capture Neutrophil transmigration events with invasive structures like
podosome/invadopode through endothelial cells (HUVECs)using TEM.

For that i fixed neutrophils during course of transmigration on HUVEC monolayer that were grown on
ALCAR Membrane, ( As was getting hard to separate monolayer from glass coverslip after epon
embedding. We are trying to cut these perpendicular to monolayer to get neutrophil-HUVEC interaction
during extravasation. (these event are rare 5-8 leukocyte on HUVEC on each perpendicular field in
ultrathin sections)

But after many try i am not getting these events under TEM. here i am describing what i am doing--

After embedding in epon Lx112, monlayers were separated from ALCAR Membrane and were re-embedded in
epon to cover monolayer--- But during ultracut thin section-- both epon part showing separation and
folding at the surface where leukocyte- HUVEC interacted when observed under transmission EM. then
i tried without re-embedding, but still folding issue led to failure to capture any of these events.

We can see many neutrophils on HUVEC monolayer on embedded blocks under light microscope and thick
section, i then used formvar-copper coated 75 mess grids to get more area to visualize and support
for sections, these coating on grids are so fragile, after so many try i standardized for section
collection and Uranyl acetate staining on these, but still not getting events of interest.(as i was
cutting section to get large monolayer area - so i thought that may be during section transfer with
loop its not staying in center of grids or covered on grid net).

So now i started cutting a small area of monolayer with 4-6 neutrophil events per section as
identified with thick sectioning 1uM --from block with good pyramid to get ribbons of section, so
that i can have multiple sections on single grid.

But somehow i am not getting ultrathin section 100nm from these block, i doubt its due to block
hardness is compromised with small area to cut. i dont know what to try next.

it would be really good, if re-embedding would work without section separation so that HUVEC
monolayar with leukocytes remain in middle of section and thus no problem with folding at borders....

I am looking forward for your help and suggestions and let me know what should i do ....

thanks

Sachin

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Hi, Sachin-

I regularly section cells grown on, fixed, and embedded in LX112 in plastic
Petri dishes. Once I manage to peel off the dish (this is the hard part), I do
not try to re-embed. I just cut the monolayer with the edge on one side. I
collect them on copper single-slot (1 x 2 mm) grids coated with Formvar (I coat
my own). This way I can see all the cells along a 2 mm length, except where
they fold. If you grow them on Aclar, can you peel off the Aclar? Or even
section the Aclar, but without re-embedding? And you shouldn't care if the
Aclar peels off while sectioning if you can pick them up on slot grids.

Aloha,
Tina

} I am trying to capture Neutrophil transmigration events with invasive
} structures like
} podosome/invadopode through endothelial cells (HUVECs)using TEM.
}
} For that i fixed neutrophils during course of transmigration on HUVEC
} monolayer that were grown on
} ALCAR Membrane, ( As was getting hard to separate monolayer from glass
} coverslip after epon
} embedding. We are trying to cut these perpendicular to monolayer to get
} neutrophil-HUVEC interaction
} during extravasation. (these event are rare 5-8 leukocyte on HUVEC on each
} perpendicular field in
} ultrathin sections)
}
} But after many try i am not getting these events under TEM. here i am
} describing what i am doing--
}
} After embedding in epon Lx112, monlayers were separated from ALCAR Membrane
} and were re-embedded in
} epon to cover monolayer--- But during ultracut thin section-- both epon part
} showing separation and
} folding at the surface where leukocyte- HUVEC interacted when observed under
} transmission EM. then
} i tried without re-embedding, but still folding issue led to failure to
} capture any of these events.
}
} We can see many neutrophils on HUVEC monolayer on embedded blocks under light
} microscope and thick
} section, i then used formvar-copper coated 75 mess grids to get more area to
} visualize and support
} for sections, these coating on grids are so fragile, after so many try i
} standardized for section
} collection and Uranyl acetate staining on these, but still not getting events
} of interest.(as i was
} cutting section to get large monolayer area - so i thought that may be during
} section transfer with
} loop its not staying in center of grids or covered on grid net).
}
} So now i started cutting a small area of monolayer with 4-6 neutrophil events
} per section as
} identified with thick sectioning 1uM --from block with good pyramid to get
} ribbons of section, so
} that i can have multiple sections on single grid.
}
} But somehow i am not getting ultrathin section 100nm from these block, i
} doubt its due to block
} hardness is compromised with small area to cut. i dont know what to try
} next.
}
}
} it would be really good, if re-embedding would work without section
} separation so that HUVEC
} monolayar with leukocytes remain in middle of section and thus no problem
} with folding at borders....
}
} I am looking forward for your help and suggestions and let me know what
} should i do ....
}
} thanks
}
} Sachin
}
}
}
}
}
}
}
}
}
}
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} Listserver Email Form V - 20120416
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} the Microscopy Listserver NO-REPLY forwarding
} system. You should send a new message to
}
} Microscopy-at-Microscopy.com
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}
} ==============================Original Headers==============================
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}

****************************************************************************
* 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*
****************************************************************************

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Dear All,

I would like to announce

Preliminary programme for the inaugural UBC 3D Correlative Light and
Electron Microscopy course

UBC Bioimaging Facility

UBC Centre for Highthroughput Phenogenomics

This course is designed for life science researchers/microscopists who
are looking to learn 3D Correlative Microscopy. Our instructors will
guide you with a series of lectures and practical demonstrations. We
will introduce a sample (or two) at the beginning of the course and take
that one sample through microCT, confocal/2P, dual beam FESEM and TEM
tomography and into advanced 3D image processing. Sample preparation
will include both chemical fixation as well as high pressure freezing.

Synopsis:

When: June 14-17, 2013
Where: UBC Bioimaging Facility and UBC Centre for High-Throughput
Phenogenomics, University of British Columbia, Vancouver, BC. Canada
Support: Systems for Research (SFR), FEI Company, Edge Scientific,
Leica Microsystems, Olympus Canada, TILL Photonics and VSG
Who: 10-12 students, preferably life sciences PhD students, Post
docs, microscopy facility managers/directors
Course fee: $2500 CDN ($1400 for UBC researchers) and will include 5
nights accommodation, breakfast, lunch and one dinner.

For more information contact Garnet Martens at garnet.martens-at-botany.ubc.ca.

Preexisting knowledge of both light and electron microscopy necessary.

Lectures will include:

Principles of CLEM
CLEM methods
microCT for CLEM
Dual beam SEM for CLEM
Tomography
CryoEM sample preparation
Confocal/2P for CLEM
Image processing and analysis

Practical sessions:

microCT scanning
confocal/2p imaging
sample handling for both chemical fixation and high pressure freezing
FIB/SEM
Tomography
Image processing and analysis

Instructors:

Elizabeth Fischer, Rocky Mountain Labs, Hamilton, MT
Vinod Nair, Rocky Mountain Labs, Hamilton, MT
William J. Rice, New York Structural Biology Center
Kim Rensing, Leica Microsystems
Richard Gursky, FEI Company
Cliff Matheson, FEI Company
Christian Wietholt, VSG
TILL Photonics
Farid Jalali, Olympus Canada
Nancy Ford, UBC Centre for High-Throughput Phenogenomics
Gethin Owen, UBC Centre for High-Throughput Phenogenomics
Kevin Hodgson, UBC Bioimaging Facility
Derrick Horne, UBC Bioimaging Facility
Brad Ross, UBC Bioimaging Facility
Garnet Martens, UBC Bioimaging Facility

--
William J. Rice, Ph.D.
Senior Scientist, EM Facility Manager
New York Structural Biology Center
89 Convent Avenue, NY, NY 10027

==============================Original Headers==============================
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From menace-at-cogeco.ca Tue May 14 10:29:36 2013
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Dear Listers,
we're going to install a brand new SEM with Schottky emission. We were
thinking of position it on a base placed into a hole dug in the floor
expressly to decouple the base from building.

On the other hand we discovered that a similar base built here some 10
years ago is not isolated that much. This base was built by digging a
hole in the floor, than filling the bottom part of the hole with gravel
and finally putting the concrete base on it. I think (not sure) the gap
between the sides of the base and the hole was filled with sand or
gravel. Finally, the same gap at floor level was filled with a rubber
expansion joint.

Do you have any better recipe? Or different solutions?
Any comment or suggestion would be greatly appreciated!
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
"Nota automatica aggiunta dal sistema di posta
Il 5 per mille per sostenere i giovani ricercatori di Ca' Foscari.
E' un atto volontario, non costa nulla e non sostituisce l'8 per mille.
Scegli Ca' Foscari: codice fiscale 80007720271
Please note that the above message is addressed only to individuals filing
Italian income tax returns. -- "

==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
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As I understand it the idea isolation system should not contact anything but bed rock. Just a concrete pillar sitting on bed rock with the walls of reinforced shaft just inches away. In some places that's impossible. No bed rock or too swampy. There are air shock mounts which "float" the scope on a cushion of air.

Don't forget the other source of vibrations, air and water lines can transmit vibrations or acoustic vibrations from compressors, I can't imagine all the possible sources. Take a look at your facility and then see what's possible.

Consider what the vibration footprint might look like in 5 years and plan for that as well.

Good luck

-----Original Message-----
X-from: dcristofori-at-unive.it [mailto:dcristofori-at-unive.it]
Sent: Wednesday, May 15, 2013 5:26 AM
To: Frank Karl

Dear Listers,
we're going to install a brand new SEM with Schottky emission. We were
thinking of position it on a base placed into a hole dug in the floor
expressly to decouple the base from building.

On the other hand we discovered that a similar base built here some 10
years ago is not isolated that much. This base was built by digging a
hole in the floor, than filling the bottom part of the hole with gravel
and finally putting the concrete base on it. I think (not sure) the gap
between the sides of the base and the hole was filled with sand or
gravel. Finally, the same gap at floor level was filled with a rubber
expansion joint.

Do you have any better recipe? Or different solutions?
Any comment or suggestion would be greatly appreciated!
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Universit� Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
"Nota automatica aggiunta dal sistema di posta
Il 5 per mille per sostenere i giovani ricercatori di Ca' Foscari.
E' un atto volontario, non costa nulla e non sostituisce l'8 per mille.
Scegli Ca' Foscari: codice fiscale 80007720271
Please note that the above message is addressed only to individuals filing
Italian income tax returns. -- "

==============================Original Headers==============================
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This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
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Hi,
IF a problem is seen, I suggest you look also into active vibration
isolation tables. These can be provided to complement the passive system
your SEM undoubtedly has. From recent research I've done, it can be quite an
undertaking to do a proper foundation isolation project. For a dedicated
facility it is done - although costly. A vibration consultant told me
recently that he also did not measure significant improvements from homemade
approaches he's inspected. Does your site meet the manufacturer's vibration
specification?

I have managed plenty of sub-nm resolution instruments on regular concrete
slabs with no isolation other than having enough distance from neighbors.

Good luck!
Larry Scipioni

-----Original Message-----
X-from: dcristofori-at-unive.it [mailto:dcristofori-at-unive.it]
Sent: Wednesday, May 15, 2013 5:28 AM
To: LES-at-ZSGENETICS.COM

Dear Listers,
we're going to install a brand new SEM with Schottky emission. We were
thinking of position it on a base placed into a hole dug in the floor
expressly to decouple the base from building.

On the other hand we discovered that a similar base built here some 10 years
ago is not isolated that much. This base was built by digging a hole in the
floor, than filling the bottom part of the hole with gravel and finally
putting the concrete base on it. I think (not sure) the gap between the
sides of the base and the hole was filled with sand or gravel. Finally, the
same gap at floor level was filled with a rubber expansion joint.

Do you have any better recipe? Or different solutions?
Any comment or suggestion would be greatly appreciated!
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi Via Torino, 155b I-30172
Mestre (VE) Italy ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ "Nota
automatica aggiunta dal sistema di posta Il 5 per mille per sostenere i
giovani ricercatori di Ca' Foscari.
E' un atto volontario, non costa nulla e non sostituisce l'8 per mille.
Scegli Ca' Foscari: codice fiscale 80007720271 Please note that the above
message is addressed only to individuals filing Italian income tax returns.
-- "

==============================Original Headers==============================
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17, 23 -- From les-at-zsgenetics.com Wed May 15 07:30:40 2013
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Disclaimer � I work for Integrated Dynamics Engineering (IDE) who is a
global provider of Vibration, EMI, and Acoustic Isolation products
specifically designed for the Microscopy market.

Davide,

There are many different solutions (Passive and Active) that are available
to you to create a vibration free floor base. If you want to contact me
directly, please e-mail me at peter.wilson-at-ideworld.com and I would be
happy to provide you with some of our white papers on projects completed at
Microscopy laboratories around the world.

Sincerely,

Peter J. Wilson
Integrated Dynamics Engineering
68 Mazzeo Drive
Randolph, MA 02368
office:781-300-6545
fax:781-326-3004
mobile:978-835-2428

-----Original Message-----
X-from: les-at-zsgenetics.com [mailto:les-at-zsgenetics.com]
Sent: Wednesday, May 15, 2013 8:38 AM
To: peter.wilson-at-ideworld.com

Hi,
IF a problem is seen, I suggest you look also into active vibration
isolation tables. These can be provided to complement the passive system
your SEM undoubtedly has. From recent research I've done, it can be quite an
undertaking to do a proper foundation isolation project. For a dedicated
facility it is done - although costly. A vibration consultant told me
recently that he also did not measure significant improvements from homemade
approaches he's inspected. Does your site meet the manufacturer's vibration
specification?

I have managed plenty of sub-nm resolution instruments on regular concrete
slabs with no isolation other than having enough distance from neighbors.

Good luck!
Larry Scipioni

-----Original Message-----
X-from: dcristofori-at-unive.it [mailto:dcristofori-at-unive.it]
Sent: Wednesday, May 15, 2013 5:28 AM
To: LES-at-ZSGENETICS.COM

Dear Listers,
we're going to install a brand new SEM with Schottky emission. We were
thinking of position it on a base placed into a hole dug in the floor
expressly to decouple the base from building.

On the other hand we discovered that a similar base built here some 10 years
ago is not isolated that much. This base was built by digging a hole in the
floor, than filling the bottom part of the hole with gravel and finally
putting the concrete base on it. I think (not sure) the gap between the
sides of the base and the hole was filled with sand or gravel. Finally, the
same gap at floor level was filled with a rubber expansion joint.

Do you have any better recipe? Or different solutions?
Any comment or suggestion would be greatly appreciated!
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi Via Torino, 155b I-30172
Mestre (VE) Italy ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ "Nota
automatica aggiunta dal sistema di posta Il 5 per mille per sostenere i
giovani ricercatori di Ca' Foscari.
E' un atto volontario, non costa nulla e non sostituisce l'8 per mille.
Scegli Ca' Foscari: codice fiscale 80007720271 Please note that the above
message is addressed only to individuals filing Italian income tax returns.
-- "

==============================Original Headers==============================
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==============================Original Headers==============================
17, 23 -- From les-at-zsgenetics.com Wed May 15 07:30:40 2013
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isolating floor bases
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Dear List Members,

I would like to announce a post-doctoral position in Materials Characterization by Transmission Electron Microscopy at University of Thessaloniki, Greece, for 18 months starting from August 1st, 2013.

The Electron Microscopy Laboratory of the Physics Department of the Aristotle University of Thessaloniki is seeking a high-qualified and motivated post-doctoral fellow with an interest in microstructural characterization of advanced materials, using Transmission Electron Microscopy (TEM).
The position is a Marie Curie fellowship funded by the European Union in the framework of the FP7 ITN project "NetFISiC" (http://www.netfisic.eu). It is an 18-month fixed-duration position, starting August 1st, 2013 and ending January 31st, 2015.
The laboratory uses microscopy techniques for atomic and nanoscale materials characterization by TEM.
Position Requirements:
The successful candidate should have a PhD in Materials Science or Physics. But the position also has a very specific timeframe: the candidate, at August 1st, 2013, should not have more than five years of research experience from the day of obtaining the Masters degree. Extended TEM experience and deep knowledge of digital image analysis and simulations using specialized software is also mandatory.
Gross salary: 4465 € per month.
Contact person: Professor E.K. Polychroniadis, polychr-at-auth.gr

More information can be found at:
http://ec.europa.eu/euraxess/index.cfm/jobs/jobDetails/33861513

Best regards,
E.K. Polychroniadis

Prof. E.K. Polychroniadis
Department of Physics
Aristotle University of Thessaloniki
Thessaloniki 54124, Greece
Tel.: +30.2310.998163
Fax: +30.2310.998241
e-mail: polychr-at-auth.gr

==============================Original Headers==============================
11, 24 -- From polychr-at-auth.gr Thu May 16 17:10:19 2013
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From noreply-at-touchingdeco.com Thu May 16 18:12:37 2013
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Davide;
After years of fighting with malfunctioning active systems at Intel, I would say avoid them like the plague. The one I had needed to be "tuned" periodically because it was not truly active, and had to be set close to the frequencies in the floor. Unfortunately, the tuning just would not hold and it would go into oscillation, which would shake the microscope violently. After 8 years, it failed totally, and we had to purchase a new one. To repair the old one, it would have to be removed from under the microscope, shipped to Europe, debugged for $250 per hour, repaired for labor and parts, and returned with minimal warranty. Meanwhile the microscope would have been unusable for all that time, all of which was unacceptable to Intel. We also paid for the expert to fly out from Germany to install and tune it, and it took three days to get it tuned so that it was not oscillating the next morning. He stopped by and tuned it again on the way to the airport, and fortunately that tune held for a few years, because help was far away.

John Mardinly
ASU

-----Original Message-----
X-from: les-at-zsgenetics.com [mailto:les-at-zsgenetics.com]
Sent: Wednesday, May 15, 2013 5:39 AM
To: John Mardinly

Hi,
IF a problem is seen, I suggest you look also into active vibration isolation tables. These can be provided to complement the passive system your SEM undoubtedly has. From recent research I've done, it can be quite an undertaking to do a proper foundation isolation project. For a dedicated facility it is done - although costly. A vibration consultant told me recently that he also did not measure significant improvements from homemade approaches he's inspected. Does your site meet the manufacturer's vibration specification?

I have managed plenty of sub-nm resolution instruments on regular concrete slabs with no isolation other than having enough distance from neighbors.

Good luck!
Larry Scipioni

-----Original Message-----
X-from: dcristofori-at-unive.it [mailto:dcristofori-at-unive.it]
Sent: Wednesday, May 15, 2013 5:28 AM
To: LES-at-ZSGENETICS.COM

Dear Listers,
we're going to install a brand new SEM with Schottky emission. We were
thinking of position it on a base placed into a hole dug in the floor
expressly to decouple the base from building.

On the other hand we discovered that a similar base built here some 10 years
ago is not isolated that much. This base was built by digging a hole in the
floor, than filling the bottom part of the hole with gravel and finally
putting the concrete base on it. I think (not sure) the gap between the
sides of the base and the hole was filled with sand or gravel. Finally, the
same gap at floor level was filled with a rubber expansion joint.

Do you have any better recipe? Or different solutions?
Any comment or suggestion would be greatly appreciated!
Thanks

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Università Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi Via Torino, 155b I-30172
Mestre (VE) Italy ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ "Nota
automatica aggiunta dal sistema di posta Il 5 per mille per sostenere i
giovani ricercatori di Ca' Foscari.
E' un atto volontario, non costa nulla e non sostituisce l'8 per mille.
Scegli Ca' Foscari: codice fiscale 80007720271 Please note that the above
message is addressed only to individuals filing Italian income tax returns.
-- "

==============================Original Headers==============================
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Davide

On the other hand I believe we have a system from the same European
manufacturer that worked well for 13 years no need for retuning.

Alan

On 5/16/2013 7:13 PM, John.Mardinly-at-asu.edu wrote:
} ----------------------------------------------------------------------------
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} Davide;
} After years of fighting with malfunctioning active systems at Intel, I would say avoid them like the plague. The one I had needed to be "tuned" periodically because it was not truly active, and had to be set close to the frequencies in the floor. Unfortunately, the tuning just would not hold and it would go into oscillation, which would shake the microscope violently. After 8 years, it failed totally, and we had to purchase a new one. To repair the old one, it would have to be removed from under the microscope, shipped to Europe, debugged for $250 per hour, repaired for labor and parts, and returned with minimal warranty. Meanwhile the microscope would have been unusable for all that time, all of which was unacceptable to Intel. We also paid for the expert to fly out from Germany to install and tune it, and it took three days to get it tuned so that it was not oscillating the next morning. He stopped by and tuned it again on the way to the airport, and fortunately that !
}
} tune held for a few years, because help was far away.
}
} John Mardinly
} ASU
}
}
} -----Original Message-----
} X-from: les-at-zsgenetics.com [mailto:les-at-zsgenetics.com]
} Sent: Wednesday, May 15, 2013 5:39 AM
} To: John Mardinly
} Subject: [Microscopy] RE: SEM: buildiing strategies for vibration isolating floor bases
}
}
}
}
} ----------------------------------------------------------------------------
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} On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} ----------------------------------------------------------------------------
}
} Hi,
} IF a problem is seen, I suggest you look also into active vibration isolation tables. These can be provided to complement the passive system your SEM undoubtedly has. From recent research I've done, it can be quite an undertaking to do a proper foundation isolation project. For a dedicated facility it is done - although costly. A vibration consultant told me recently that he also did not measure significant improvements from homemade approaches he's inspected. Does your site meet the manufacturer's vibration specification?
}
} I have managed plenty of sub-nm resolution instruments on regular concrete slabs with no isolation other than having enough distance from neighbors.
}
} Good luck!
} Larry Scipioni
}
} -----Original Message-----
} X-from: dcristofori-at-unive.it [mailto:dcristofori-at-unive.it]
} Sent: Wednesday, May 15, 2013 5:28 AM
} To: LES-at-ZSGENETICS.COM
} Subject: [Microscopy] SEM: buildiing strategies for vibration isolating floor bases
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Dear Listers,
} we're going to install a brand new SEM with Schottky emission. We were
} thinking of position it on a base placed into a hole dug in the floor
} expressly to decouple the base from building.
}
} On the other hand we discovered that a similar base built here some 10 years
} ago is not isolated that much. This base was built by digging a hole in the
} floor, than filling the bottom part of the hole with gravel and finally
} putting the concrete base on it. I think (not sure) the gap between the
} sides of the base and the hole was filled with sand or gravel. Finally, the
} same gap at floor level was filled with a rubber expansion joint.
}
} Do you have any better recipe? Or different solutions?
} Any comment or suggestion would be greatly appreciated!
} Thanks
}
} Davide
}
}
} ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ Davide Cristofori
}
} direct phone = 041.234.67.26
} e-mail = dcristofori[at]unive[dot]it
}
} Universit� Ca' Foscari Venezia
} Dipartimento di Sc. Molecolari e Nanosistemi Via Torino, 155b I-30172
} Mestre (VE) Italy ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~ "Nota
} automatica aggiunta dal sistema di posta Il 5 per mille per sostenere i
} giovani ricercatori di Ca' Foscari.
} E' un atto volontario, non costa nulla e non sostituisce l'8 per mille.
} Scegli Ca' Foscari: codice fiscale 80007720271 Please note that the above
} message is addressed only to individuals filing Italian income tax returns.
} -- "
}
}

--
Alan W Nicholls, PhD
Associate Director, RRC
Director of Research Service Facility (Electron Microscopy)
Research Resources Center - East (M/C 337)
Room 110 Science and Engineering South Building
The University of Illinois at Chicago
845 West Taylor St
Chicago, IL 60607-7058

Tel: 312 996 1227
Office: Room 110
nicholls-at-uic.edu
Web Site: http://www.rrc.uic.edu/ems
Wiki site https://wiki.rrc.uic.edu/wiki/EMS

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Email: Mitch.Kupferman-at-usa.dupont.com
Name: Mitchell Kupferman

Organization: DuPont-Teijin Films

Title-Subject: [Filtered] LEO 435 VP software conflict

Message: Good Morning!

I have a LEO 435VP SEM in my lab which up until recently has been running
well. Bringing the instrument up from a routine shutdown, I found that I
no longer had the ability to access the variable pressure software without
shutting down the software that operates the stage. The software that
controls the variable pressure has always been in conflict with the stage
movement/control software, but the one has never stopped the other from
working properly. After a few reboots, the problem persisted and it seems
that I can no longer operate the stage while in VP. The error message
that appears on screen is: "VP comm error 6".
The instrument is rather old (OS windows 3.1) and I'd like to avoid
removing the software and reinstalling it as I'm afraid I may lose more
significant functions. Has any with this instrument seen this type of
error before, and if so, how were you able to solve it?

Thanks for any help.

Mitchell Kupferman
DuPont Teijin Films
Analytical Chemist
Hopewell Site
3600 Discovery Dr.,
Chester VA 23836
804.530.9383 (o)

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I've used TMC's Stacis active vibration cancelling system with excellent results on the 3rd floor of a 50-year-old building that vibrates at 15Hz constantly. This frequency seemed to be the resonant frequency of the stage of my large chamber SEM and was causing plenty of trouble.

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}
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Dear Listers,

May I have your help with the below issue:

I am trying to take a TEM tilt series of MgO placed on 300 mesh Cu
Holey carbon grids,
at the beginning everything was great but after the 10th image was
acquired, I noticed that the sample started to change ( picture below)
presumably because the interaction between the beam and the Carbon
grid?!
my question is: are there anything that can be done to stop this
interaction (may be by prepositioning or using other Grid)?
(note: I thought of cleaning the sample with plasma, but unfortunately
my sample is very thin)

(picture on this link: https://sites.google.com/site/alaaalfeef/home/Tv14.bmp)

Thank you

very much for your help
--

Ala' Afeef,
PhD Scholar - Glasgow University

==============================Original Headers==============================
7, 27 -- From alaa.afeef-at-gmail.com Mon May 20 06:57:49 2013
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Hi Ala' Afeef,

I don't see a before and after, so I don't know what the changes are, but it looks like you may have some carbon contamination at the very least.

Even when you can't plasma clean a particular sample, plasma cleaning the holder (including the nut and washer) still gives a noticeable improvement.

If you send a before and after shot, or better, a link to the whole tilt sequence, maybe we can make a more complete comment.

Cheers,

Zack Gainsforth
Space Sciences Laboratory, UC Berkeley
7 Gauss Way
Berkeley, CA 94720
work: 510-642-9733
cell: 626-437-9186
zackg-at-ssl.berkeley.edu

On May 20, 2013, at 5:09 AM, alaa.afeef-at-gmail.com wrote:

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Dear Listers,

May I have your help with the below issue:

I am trying to take a TEM tilt series of MgO placed on 300 mesh Cu
Holey carbon grids,
at the beginning everything was great but after the 10th image was
acquired, I noticed that the sample started to change ( picture below)
presumably because the interaction between the beam and the Carbon
grid?!
my question is: are there anything that can be done to stop this
interaction (may be by prepositioning or using other Grid)?
(note: I thought of cleaning the sample with plasma, but unfortunately
my sample is very thin)

(picture on this link: https://sites.google.com/site/alaaalfeef/home/Tv14.bmp)

Thank you

very much for your help
--

Ala' Afeef,
PhD Scholar - Glasgow University

==============================Original Headers==============================
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Hi Ala' Afeef,

I agree with Zack, looks very much like carbon contamination - note this
is not modification of your carbon substrate but a deposition of carbon
from the cracking of free hydrocarbons under e-beam heating.

You can also see that those MgO cubes away from your area of interest
are still clean, while the one you are have been looking at has a thick
coating (and the one next door!) with the carbon film around darkening
from a thinner coating (the difference in heating rates and/or
conductivity between C and MgO may be to blame for this difference).

I would recommend testing your grids before depositing MgO cubes on them
(presumably by waiving the grids through burning Mg), to verify their
cleanliness. You can try to remove free hydrocarbon by baking the
grid/film in vacuum, or a short cut placing them on an incandescent
light bulb for a few seconds! You can also plasma clean carbon films,
but only for a very short time (30 seconds or so depending on your
plasma cleaner). Some reports state that a H/O plasma in gentler on
films than traditional Ar/O. Have a play with different times and see
what works... It may also be possible it's your holder and/or microscope
that is the source of contamination.. (but fingers crossed it is not)

Good luck.

Matthew

} Dear Listers,
}
} May I have your help with the below issue:
}
} I am trying to take a TEM tilt series of MgO placed on 300 mesh Cu
} Holey carbon grids,
} at the beginning everything was great but after the 10th image was
} acquired, I noticed that the sample started to change ( picture below)
} presumably because the interaction between the beam and the Carbon
} grid?!
} my question is: are there anything that can be done to stop this
} interaction (may be by prepositioning or using other Grid)?
} (note: I thought of cleaning the sample with plasma, but unfortunately
} my sample is very thin)
}
} (picture on this link: https://sites.google.com/site/alaaalfeef/home/Tv14.bmp)
}
} Thank you
}
} very much for your help
} --
}
} Ala' Afeef,
} PhD Scholar - Glasgow University

--
Dr M.Weyland, Senior Lecturer & Titan Manager
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Title-Subject: [Filtered] Schematic for Pfeiffer TCP 300 pump Controller

Message: I'm looking for a schematic so I can troubleshoot. My pump won't statart.

Thanks

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Lee, connection diagram of TCP 300 controller and some bits of
information on internals are in the standard manual:

http://www.idealvac.com/files/manualsII/Pfeiffer_Balzers_TCP-300_Controller.pdf

Troubleshooting on component level typically becomes an exercise in
electronics reverse-engineering, which is most certainly do-able - I've
done it quite a few times - but more often then not purchasing
second-hand controller turns out to be more cost- and time- effective...

Good luck! :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

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Okay.
I must nuts. We have an Amray 1803I with turbo pump and an ion getter. After we change a W filament it takes about 4-5 hours to pump down to a usable vacuum. We have to wrap on the getter, open and close valves trying to urge the system on to a good vacuum, chanting does seem to help. We define good as the alarm doesn't go off and we can work. We're not sure where the alarm position is set. Of course Amray is gone, but not forgotten

I've used 5 different SEM in my 30 years as a Microscopist. I don't ever remember changing a filament taking this long, but I never had an ion getter on a SEM, only on my TEMs. Do any other Amray users have any comments or observations?

Thanks............

Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Hello,

I'm experiencing very large astigmatism at lower accelerating
voltages in our Jeol SEM. The only voltage at which I can
compensate for and obtain decent quality picture is 25kV.

Any ideas why?

Thanks,

Laci

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Hi Laci,

Any chance your sample is magnetic? That would likely have a greater effect at lower kV's.

Cheers,
Randy

-----Original Message-----
X-from: jflaci-at-ms.sapientia.ro [mailto:jflaci-at-ms.sapientia.ro]
Sent: Tuesday, May 21, 2013 3:06 PM
To: Tindall, Randall D.

Hello,

I'm experiencing very large astigmatism at lower accelerating voltages in our Jeol SEM. The only voltage at which I can compensate for and obtain decent quality picture is 25kV.

Any ideas why?

Thanks,

Laci

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Hi Laci

What you mention is a common problem, there is a high degree of charge on
the components requiring a excessive level of astigmatism correction.
Basically the column is contaminated and by moving to a higher accelerating
voltage you enable the contamination to be penetrated enabling an earth to
be achieved and an astigmatism correction to be made. At lower accelerating
voltages the beam does not penetrate the contamination thus there is a high
degree of astigmatism, too high for the stigmator coils to compensate.

Clean the column and if you have a service technician who maintains the
instrument make sure they cover all aspects of operation during a
maintenance procedure. Running the instrument at a high accelerating
voltage checks out the gun; fine. Running the instrument at a very low
accelerating voltage checks out the column and this is the more important
check. Thus a 30kV test picture for the gun stability and as a resolution
check, but a 2kV test picture for a real check of column cleanliness!

Enjoy

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: jflaci-at-ms.sapientia.ro [mailto:jflaci-at-ms.sapientia.ro]
Sent: 21 May 2013 20:56
To: protrain-at-emcourses.com

Hello,

I'm experiencing very large astigmatism at lower accelerating voltages in
our Jeol SEM. The only voltage at which I can compensate for and obtain
decent quality picture is 25kV.

Any ideas why?

Thanks,

Laci

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Frank,

Getter pumps do have a finite life. Maybe yours is reaching the end of its capacity to pump efficiently.

We used to have a microscope with a LaB6 gun and ion pump. After 10 years of operation it was slowing down.

Rick

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Okay.
I must nuts. We have an Amray 1803I with turbo pump and an ion getter. After we change a W filament it takes about 4-5 hours to pump down to a usable vacuum. We have to wrap on the getter, open and close valves trying to urge the system on to a good vacuum, chanting does seem to help. We define good as the alarm doesn't go off and we can work. We're not sure where the alarm position is set. Of course Amray is gone, but not forgotten

I've used 5 different SEM in my 30 years as a Microscopist. I don't ever remember changing a filament taking this long, but I never had an ion getter on a SEM, only on my TEMs. Do any other Amray users have any comments or observations?

Thanks............

Frank

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On my FEI Quanta ESEM, that kind of trouble tells me to clean the last PLA first.

Hope it's that simple.

Fred Monson

Frederick C Monson, PhD
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pannsylvania
West Chester, PA, 1938
610-738-0437
fmonson-at-wcupa.edu
________________________________________
X-from: tindallr-at-missouri.edu [tindallr-at-missouri.edu]
Sent: Tuesday, May 21, 2013 4:18 PM
To: Monson, Frederick

Hi Laci,

Any chance your sample is magnetic? That would likely have a greater effect at lower kV's.

Cheers,
Randy

-----Original Message-----
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Sent: Tuesday, May 21, 2013 3:06 PM
To: Tindall, Randall D.

Hello,

I'm experiencing very large astigmatism at lower accelerating voltages in our Jeol SEM. The only voltage at which I can compensate for and obtain decent quality picture is 25kV.

Any ideas why?

Thanks,

Laci

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Email: tomw-at-uidaho.edu
Name: Tom Williams

Organization: University of Idaho

Title-Subject: [Filtered] FOM-FIG "Lunch With a Manager" -at- MM 2013

Message: Greetings,
Supporting and cultivating the next generation of Microscopists is an important mission of MSA and
the Facilities Operations and Management FIG. At the 2013 MM Conference in Indianapolis the FOM-FIG
is pleased to sponsor and host a lunch & roundtable discussion for students and postdocs considering
a career in microscopy, or managing a microscopy facility. We are inviting 20 participants to share
a lunch with a panel of veteran microscopists and managers for an open forum and discussion. If you
have or know a student, recent graduate, or postdoc who might be interested in attending I would
greatly appreciate passing along the invitation below. Registration is FREE and on a first come basis.

Thanks,
Tom Williams

An Invitation
Microscopy & Microanalysis 2013-Indianapolis
Lunch with a Manager

Sponsored by Facilities Operations &
Management-Focused Interest Group

Graduating soon and not sure what you want to do?
Maybe considering a career managing a
Microscopy Center and wondering how to land
such a great job? Or just curious what a Lab
Manager does all day? Heres your opportunity to
find out!

The MSA FOM-FIG is pleased to host a Lunch &
Roundtable forum with a panel of veteran
Facility Managers and Administrators from
Academia & Industry.

The Particulars:
Monday [5 Aug.]
Time: 12:00-1:15
Location: Rm 211 [Indiana Conv. Center]
Food & Drinks Provided*
*box lunch [w/vegetarian option]

Seating is limited to 20 participants on a first come
basis!

Deadline to register: 29 July

To Register:
Contact Tom Williams at the
University of Idaho:
tomw-at-uidaho.edu
-provide a contact email, institutional affiliation,
and any dietary accommodations or restrictions.

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Dear Colleagues,
We would like to remind you of the upcoming Yale Microscopy Workshop that
will be held June 4 -6th 2013. This year's symposia theme is Correlative
Light and Electron Microscopy. This free event is a combination of
symposia, demonstrations and technical lectures representing all imaging
modalities that this year includes several unique to CLEM.

This workshop also provides open access to functional microscopes, sample
preparation instruments and analytic software. It is the perfect
opportunity to compare instruments, get advice or try out something new.

Please see the website for free online registration, a listing of
available equipment and a schedule of events too numerous to mention here:
http://medicine.yale.edu/lab/microscopy/index
{http://microscopy.med.yale.edu/index.html}

Yale Microscopy Workshop Symposium Schedule
----------------------------
Tuesday June 4th Afternoon symposium: Correlative Light and Electron
Microscopy Part I

"Visualizing Cells and Viruses at Molecular Resolution: From Structure to
Mechanism"
Sriram Subramaniam, NIH/NCI

"Correlative (i)PALM and EM Imaging"
Gleb Shtengel, HHMI Janelia Farm

----------------------------
Wednesday June 5th Afternoon symposium: Correlative Light and Electron
Microscopy Part I

"Correlative light and electron microscopy reveals new structure in the
immunological synapse"
Michael Dustin, Skirball Institute

"Development and Application of Labeling Technologies to Propel Correlated
Multi-Scale Imaging of Cells and Tissues"
Mark Ellisman, UCSD

We look forward to seeing you there!

The organizers,
Ann Haberman (ann.haberman-at-yale.edu)
Derek Toomre (derek.toomre-at-yale.edu)
Joerg Bewersdorf (joerg.bewersdorf-at-yale.edu)
Xinran Liu (xinran.liu-at-yale.edu)

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I agree. But many use an alternative, much easier method to solve the
problem - an in-chamber downstream plasma cleaner. To be sure this doesn't
clean up right inside the column, but most of the problem is at the sharp
end, where the plasma can reach, for 2 reasons: (1) That is mostly where the
contamination condenses (2) that is where the beam energy is low. (The
method is a high energy flight tube followed by beam-deceleration.) So a
method that I know some have found to be effective is to mount an
in-SEM-chamber downstream plasma (such as that from XEI) and run this for a
few minutes a week, restoring much low-energy performance.

Ian Holton
Acutance Scientific

-----Original Message-----
X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
Sent: 21 May 2013 21:45
To: ian-at-acutance.co.uk

Hi Laci

What you mention is a common problem, there is a high degree of charge on
the components requiring a excessive level of astigmatism correction.
Basically the column is contaminated and by moving to a higher accelerating
voltage you enable the contamination to be penetrated enabling an earth to
be achieved and an astigmatism correction to be made. At lower accelerating
voltages the beam does not penetrate the contamination thus there is a high
degree of astigmatism, too high for the stigmator coils to compensate.

Clean the column and if you have a service technician who maintains the
instrument make sure they cover all aspects of operation during a
maintenance procedure. Running the instrument at a high accelerating
voltage checks out the gun; fine. Running the instrument at a very low
accelerating voltage checks out the column and this is the more important
check. Thus a 30kV test picture for the gun stability and as a resolution
check, but a 2kV test picture for a real check of column cleanliness!

Enjoy

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: jflaci-at-ms.sapientia.ro [mailto:jflaci-at-ms.sapientia.ro]
Sent: 21 May 2013 20:56
To: protrain-at-emcourses.com

Hello,

I'm experiencing very large astigmatism at lower accelerating voltages in
our Jeol SEM. The only voltage at which I can compensate for and obtain
decent quality picture is 25kV.

Any ideas why?

Thanks,

Laci

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I have a user who is looking to do some cold stage ESEM work this summer,
and she would like to purchase some of the cold stage holders before hand.

These are for an FEI ESEM: "The diameter is 0.38" and they are 0.20" high.
They are made out of stainless steel and have a slot on the sides and
bottom. This is to release any pressure that might develop during cold
temperature operation."

Commercial vendors feel free to contact me directly.

Thank you.

Richard E. Edelmann, Ph.D.
Official Program and Meeting Guide Editor for M&M 2012
Microscopy & Microanalysis

Director, Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-miamioh.edu
http://www.cami.muohio.edu

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I want to thank everyone who responded to my query on ion getter pump down times.

It appears I am somewhat of a crank and that 4 hours to pump down an Amray with a IG isn't excessive. I appreciate the links to refurbishing and replacement companies as well as instructions and suggestions.

Perspective. It's good to get fresh perspective.

Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Dear List Members,

We are pleased to invite you to the International Multidisciplinary
Microscopy Congress - INTERM 2013, which will be held on October 10-13, 2013
in Antalya, Turkey, in the Limak Limra Hotel & Resort.

The aim of the congress is to convene scientists from various branches and
discuss the latest improvements in the field of microscopy. The contents of
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present their work.

The congress proceedings are scheduled to be published under the series of
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For more information, please visit: www.interem.org

Best regards,
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Prof. E.K. Polychroniadis
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Hello Laci,

We agree with Ian Holton's possible solution to the problem. Removal of
carbon contamination from the chamber will minimize astigmatism and improve
low kV image quality.

Plasma cleaning solutions other than those offered by XEI exist. Sample
cleaning before examination removes the carbon prior to polymerization by
the electron beam. ibss Group offers an in-situ plasma cleaner that runs at
pressures compatible with TMP operation. This provides more thorough chamber
and column cleaning with fast turnaround time.

Disclaimer: the sender is President of ibss Group, Inc.

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}
} I agree. But many use an alternative, much easier method to solve the
problem - an in-chamber downstream plasma cleaner. To be sure this
} doesn't clean up right inside the column, but most of the problem is
} at
the sharp end, where the plasma can reach, for 2 reasons: (1) That
} is mostly where the contamination condenses (2) that is where the beam
energy is low. (The method is a high energy flight tube followed by
} beam-deceleration.) So a method that I know some have found to be
effective is to mount an in-SEM-chamber downstream plasma (such as
} that from XEI) and run this for a few minutes a week, restoring much
low-energy performance.
}
} Ian Holton
} Acutance Scientific
}
} -----Original Message-----
} X-from: protrain-at-emcourses.com [mailto:protrain-at-emcourses.com]
} Sent: 21 May 2013 21:45
} To: ian-at-acutance.co.uk
} Subject: [Microscopy] Large astigmatism at low kV.
}
}
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} Hi Laci
}
} What you mention is a common problem, there is a high degree of charge
} on
the components requiring a excessive level of astigmatism correction.
} Basically the column is contaminated and by moving to a higher
accelerating voltage you enable the contamination to be penetrated
} enabling an earth to be achieved and an astigmatism correction to be made.
At lower accelerating voltages the beam does not penetrate the
} contamination thus there is a high degree of astigmatism, too high for
} the
stigmator coils to compensate.
}
} Clean the column and if you have a service technician who maintains
} the
instrument make sure they cover all aspects of operation during a
} maintenance procedure. Running the instrument at a high accelerating
voltage checks out the gun; fine. Running the instrument at a very
} low accelerating voltage checks out the column and this is the more
important check. Thus a 30kV test picture for the gun stability and
} as a resolution check, but a 2kV test picture for a real check of
} column
cleanliness!
}
} Enjoy
}
} Steve
}
} Steve Chapman FRMS
} Protrain for Consultancy and Courses in Electron Microscopy
} Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
} www.emcourses.com
}
}
} -----Original Message-----
} X-from: jflaci-at-ms.sapientia.ro [mailto:jflaci-at-ms.sapientia.ro]
} Sent: 21 May 2013 20:56
} To: protrain-at-emcourses.com
} Subject: [Microscopy] Re: Large astigmatism at low kV.
}
}
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} Hello,
}
} I'm experiencing very large astigmatism at lower accelerating voltages
} in
our Jeol SEM. The only voltage at which I can compensate for and
} obtain decent quality picture is 25kV.
}
} Any ideas why?
}
} Thanks,
}
} Laci
}
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Hello,

Thank You all for the valuable input, the problem has been solved trough
a full cleanup of the lower column, where impurites have been found,
especially cracked oil. So it seems to me that I do now really have to mount
a high vacuum gauge on the scope. What would be an acceptable level
of vacuum for a 30 year old SEM, supposing we do not want to contaminate
the column when working?

Thanks,

Laci

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5, 29 -- Subject: [SOLVED] Large astigmatism at low kV.
5, 29 -- From: =?ISO-8859-1?Q?Laszl=F3_Jakab=2DFarkas?= {jflaci-at-ms.sapientia.ro}
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Email: aornelas1987-at-gmail.com
Name: Andrew

Title-Subject: [Filtered] SEM Zeiss Leo Service Company

Message: Hello,

We would like to hear about SEM servicing companies in the Northern California/Bay Area that would
be able to perform routine maintenance and PM's on a Zeiss Leo FEG-SEM. If I could hear from anyone
interested, please send messages offline.

Thank you,
Andrew

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Hi Laci,

High vacuum gauge is helpful and vacuum levels somewhere from low E-5
range down to E-7 may be reasonable, depending on the design of your
vacuum system. But to prevent contamination of the column content of
your "vacuum" (i.e. partial pressure of oils) is way more important then
ultimate pressure. It may seem counter-intuitive, but there will be way
more contamination from cracked oils in SEM chamber with ultimate
pressure E-8 Torr (very good vacuum) and main component of residual gas
mixture being mineral oil from the roughing pump, then in the SEM with
ultimate chamber pressure in low E-5 Torr range (not so great vacuum)
but main component of gas mixture being atmospheric gases.

What I am trying to say is that it is more important to keep your
chamber "dry", i.e. free of crackable hydrocarbons, then pump it down to
lowest possible vacuum level. Plasma cleaners do help here, but old
instrument would typically need thorough cleaning of entire vacuum
system. Cleaning is not difficult to do, if you have couple of days of
time and either good on-site technician or just someone who is handy
with with wrenches and vacuum flanges.

There are lots of people on this list with good vacuum expertise, so if
you describe what kind of pumping system you have in the SEM I am sure
you could get all the advice needed to dry your instrument.

Cheers :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 5/23/2013 7:37 AM, jflaci-at-ms.sapientia.ro wrote:
} ----------------------------------------------------------------------------
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} Hello,
}
}
} Thank You all for the valuable input, the problem has been solved trough
} a full cleanup of the lower column, where impurites have been found,
} especially cracked oil. So it seems to me that I do now really have to mount
} a high vacuum gauge on the scope. What would be an acceptable level
} of vacuum for a 30 year old SEM, supposing we do not want to contaminate
} the column when working?
}
} Thanks,
}
} Laci
}
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To all:

I recently used these grids to look at mice bone marrow cells, specifically megakaryocytes. In the past I had used regular mesh grids, but found that the megakaryoctes were either partially or fully obscured by the grid bars. I had beautiful sections (and megakaryocytes), but when I took the images, they were out of focus. It seemed that the formvar film (and section) moved under the beam. I haven't used these slotted grids in awhile, so not sure what the problem is and what trick is needed when viewing them under the scope.

Any help would be appreciated.
Thanks!
Peggy

Peggy Sherwood
Research Specialist, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
50 Blossom Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org

The information in this e-mail is intended only for the person to whom it is
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==============================Original Headers==============================
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Hi

Have you the option to place your sections in a scanning electron
microscope? Sit your specimen, with the grid on the bottom side, on a stub
attached at one corner by a touch of adhesive.

Run at 2 to 5kV and you should be able to view your sections without
interference from the grid bars. Reverse the image and you would not know
that you were looking at SEM as the contrast will now be the same as TEM.

Have fun!

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: MSHERWOOD-at-partners.org [mailto:MSHERWOOD-at-partners.org]
Sent: 23 May 2013 17:13
To: protrain-at-emcourses.com

To all:

I recently used these grids to look at mice bone marrow cells, specifically
megakaryocytes. In the past I had used regular mesh grids, but found that
the megakaryoctes were either partially or fully obscured by the grid bars.
I had beautiful sections (and megakaryocytes), but when I took the images,
they were out of focus. It seemed that the formvar film (and section) moved
under the beam. I haven't used these slotted grids in awhile, so not sure
what the problem is and what trick is needed when viewing them under the
scope.

Any help would be appreciated.
Thanks!
Peggy

Peggy Sherwood
Research Specialist, Photopathology
Wellman Center for Photomedicine (EDR 214) Massachusetts General Hospital
50 Blossom Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org

The information in this e-mail is intended only for the person to whom it is
addressed. If you believe this e-mail was sent to you in error and the
e-mail contains patient information, please contact the Partners Compliance
HelpLine at http://www.partners.org/complianceline . If the e-mail was sent
to you in error but does not contain patient information, please contact the
sender and properly dispose of the e-mail.

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To all:

I had a lot of good suggestions. Most people recommended exposing the grid/film at low mags to the beam to "stabilize" the film or simply waiting a few seconds (or minutes!) to do the same. When I took the images, the film/section seemed to be stable, but I will try again.

I don't have access to an SEM, but thanks Steve for that suggestion.

I don't cast my own films, I buy the slotted grids already coated with formvar and formvar/carbon. I did not use the carbon coated grids because it is just another layer to focus through. (When I routinely section on mesh grids, I use no coating at all for that very reason.

I will try and scope these same grids if they are still OK.

Again, many thanks to all who replied.

Peggy

Peggy Sherwood
Research Specialist, Photopathology
Wellman Center for Photomedicine (EDR 214)
Massachusetts General Hospital
50 Blossom Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
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Peggy,

Some of the reasons for movement of the section on slotted grids are
either a hole somewhere in the film or that the grids need to be
conditioned especially if the section is a bit thick.

If in fact the cause of your problem is movement and not focus it may be
advisable to look at a structure that is in the corner of your viewing
area both before and after taking the image. If the structure is moving
during the time of capture you will see how far it has moved - proof that
it is movement and not focus that is your problem.

If you have a hole, you can attempt to move the grid in the opposite
direction of the movement and immediately take the image. This works
sometimes but not always and is dependent on the amount of movement. One
can take advantage of that short interval between the grid moving in one
direction while the section is moving counter to it.

To counteract the change in size of the section it may be necessary to
expose the whole section to a reduced beam at a lower magnification. It
will be like scanning with the beam until the whole section has been
exposed. If there are folds present (and there are usually some found in
sections picked up on coated grids) it may be advisable to simply hit the
fold with the beam until it shrinks. I have found that going from the
pointed end towards the wide end works best.
Then go back to the area of interest with the chosen magnification and
normal beam intensity.

I have also tried carbon coating the formvar grids to give them extra
support. This works if the formvar is a bit thin.

Recently I have picked up a few 50 mesh coated grids at the same time as
the slotted ones in case I have a problem with the slotted ones. This
gives a large area and if one square has a hole the others are usually
fine hence there is usually no need to re-section.

Pat

Patricia Stranen Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E � Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-nhlbi.nih.gov

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. This message is not confidential and can be freely
shared and reproduced.

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Hi Peggy,

I don't know what kind of of camera you're using, but this happens sometimes with our Gatan digital system. It seems that often when we take a picture the beam is blanked by the shutter, then comes on for the picture. Apparently the film/resin can react quickly to the removal of the beam (thermal expansion and contraction?), causing it to move during the exposure when the beam comes back on. We can sometimes solve this by switching the camera to use the post-, rather than pre-specimen shutter in the advanced settings for the view and acquire windows. At other times, we manually shorten the exposure time to 0.5 second or even less.

We have even found that sometimes the software seems to set the acquire window for one shutter and the view window for the other, which often results in streaky images during viewing.

If all else fails, we can save the viewing window directly to a file, rather than telling the system to actually acquire a final image. This is a smaller file and what you see is what you get, but way better than nothing.

Just a couple thoughts. Good luck with this.

Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211

Phone: 573-884-5383
Email: tindallr-at-missouri.edu
Day job: www.emc.missouri.edu
Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
Blog: http://nadiasyard.com

-----Original Message-----
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Sent: Thursday, May 23, 2013 12:09 PM
To: Tindall, Randall D.

To all:

I had a lot of good suggestions. Most people recommended exposing the grid/film at low mags to the beam to "stabilize" the film or simply waiting a few seconds (or minutes!) to do the same. When I took the images, the film/section seemed to be stable, but I will try again.

I don't have access to an SEM, but thanks Steve for that suggestion.

I don't cast my own films, I buy the slotted grids already coated with formvar and formvar/carbon. I did not use the carbon coated grids because it is just another layer to focus through. (When I routinely section on mesh grids, I use no coating at all for that very reason.

I will try and scope these same grids if they are still OK.

Again, many thanks to all who replied.

Peggy

Peggy Sherwood
Research Specialist, Photopathology
Wellman Center for Photomedicine (EDR 214) Massachusetts General Hospital
50 Blossom Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org

The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail.

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Try using a carbon film. Not as susceptible to sagging and thermal stress under the beam. You will need a very high quality and fine grained film to withstand the large gap however. Alternative carbon coating the formvar to stabilize.

Scott Whittaker
Manager SEM Lab
Smithsonian Institution
National Museum of Natural History
PO Box 37012�� MRC104
Washington DC 20013-7012
202-633-0891

Welcome to our brand new facility now located in W150!

-----Original Message-----
X-from: MSHERWOOD-at-partners.org [mailto:MSHERWOOD-at-partners.org]
Sent: Thursday, May 23, 2013 12:14 PM
To: Whittaker, Scott

To all:

I recently used these grids to look at mice bone marrow cells, specifically megakaryocytes. In the past I had used regular mesh grids, but found that the megakaryoctes were either partially or fully obscured by the grid bars. I had beautiful sections (and megakaryocytes), but when I took the images, they were out of focus. It seemed that the formvar film (and section) moved under the beam. I haven't used these slotted grids in awhile, so not sure what the problem is and what trick is needed when viewing them under the scope.

Any help would be appreciated.
Thanks!
Peggy

Peggy Sherwood
Research Specialist, Photopathology
Wellman Center for Photomedicine (EDR 214) Massachusetts General Hospital
50 Blossom Street
Boston, MA 02114-2696
617-724-4839 (voice mail)
617-726-6983 (lab)
617-726-1206 (fax)
msherwood-at-partners.org

The information in this e-mail is intended only for the person to whom it is addressed. If you believe this e-mail was sent to you in error and the e-mail contains patient information, please contact the Partners Compliance HelpLine at http://www.partners.org/complianceline . If the e-mail was sent to you in error but does not contain patient information, please contact the sender and properly dispose of the e-mail.

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HI Peggy,
The large 1mm x 2mm open area of a slot grid is particularly susceptible
to drift in an EM. It is usually possible to stabilize the section and
support film by letting it sit in the beam for a few minutes. Sometimes
I spread the beam to cover a larger area and wait longer. Sometimes I
use a more focused beam and move around the section particularly
covering the edges of the grid support. If there are holes or any tears
in the support film stability is decreased and may not be possible.
Usually if you watch the specimen with binoculars or if using a digital
camera, watch the display. Any movement will blur the micrograph.
Sometimes with slow drift a short exposure time will produce an
acceptable micrograph. If none of the above works you can coat the
sections on support film with carbon to give added strength and
electrical conductivity to the sample which should eliminate or minimize
drift.
Good luck,
Larry

On 5/23/2013 9:23 AM, MSHERWOOD-at-partners.org wrote:
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} I recently used these grids to look at mice bone marrow cells, specifically megakaryocytes. In the past I had used regular mesh grids, but found that the megakaryoctes were either partially or fully obscured by the grid bars. I had beautiful sections (and megakaryocytes), but when I took the images, they were out of focus. It seemed that the formvar film (and section) moved under the beam. I haven't used these slotted grids in awhile, so not sure what the problem is and what trick is needed when viewing them under the scope.
}
} Any help would be appreciated.
} Thanks!
} Peggy
}
} Peggy Sherwood
} Research Specialist, Photopathology
} Wellman Center for Photomedicine (EDR 214)
} Massachusetts General Hospital
} 50 Blossom Street
} Boston, MA 02114-2696
} 617-724-4839 (voice mail)
} 617-726-6983 (lab)
} 617-726-1206 (fax)
} msherwood-at-partners.org
}
}
}
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--
Larry Ackerman, Specialist
Electron Microscopy Lab
Manager, Diabetes & Endocrinology Research Center Microscopy Core
UCSF, Dept. of Anatomy, Rm S657
513 Parnassus Ave., Box 0452
San Francisco, CA 94143

larry.ackerman-at-ucsf.edu

415-999-4758

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Yes, the slow heating of a large area will stop some heat/expansion
problems.

I've also seen a potential issue on systems with column shutters that are
used with the digital camera- if the grid is shielded from the beam for a
long enough time for it to cool and start to shrink, then during the
exposure it will be re-expanding. On our Gatan system, I am pretty sure
there is a time delay setting after the shutter is set to open (to allow
for it to be fully open) which you could extend into the several second
range so that things stabilise before the exposure starts, if you suspect
this might be occurring.

Ben

--
Research Support Manager
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom.
{http://www.mrc.ox.ac.uk/}

On 23/05/2013 18:05, "MSHERWOOD-at-partners.org" {MSHERWOOD-at-partners.org}
wrote:

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14, 31 -- From ben.micklem-at-pharm.ox.ac.uk Thu May 23 12:30:21 2013
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Peggy,

This is a little off the subject but you might find helpful a method for
picking up sections on single hole grids that minimizes folds, etc. It is
great for serial sections. As more people do tomography, putting sections
on single hole grids will also be very helpful in making sure the area of
interest in visible even at high tilts.
You can download this method from the RESOURCE page on my website at:

http://www.dsimagingllc.com/

Since this method works so well, I found that we could cut much wider
sections and almost cover the formvar film. This helped stabilize by
minimizing areas of formvar without requiring additional carbon coating.

The suggestions to condition the samples by sitting them under a broad
beam at low magnification also works well to minimize drift.

Debby

On 5/23/13 12:14 PM, "MSHERWOOD-at-partners.org" {MSHERWOOD-at-partners.org}
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Davide:

Coming in late to the party here. But some points. Since it sounds as
if you are actually on the lowest level and have ground below you.

We just built a new facilty within isolated floors three years ago, but
started the process before then. Yes, I had looked in to "good strategies"
and had engineers designing the renovations as well. BUT then I found
more information: (A) I found that a number of microscope manufacturers
actually have folks to help with room designs \ specs - although they do not
like to make recommendations lest they be held accountable. (B) I was
pointed in the direction of an engineering firm who specifically designed
environmental room solutions. And they first things they really pointed out
was #1 each location is unique and needs a unique solution there is no one
"good strategy", #2 when they found we were on ground they wanted to
know what the ground was really. And that got my geologist users involved
and boy we knew exactly what the ground was.

Yes, we to having read lots of individual isolation slabs with rubber gaps or
open gaps, sand filled, etc. had been thinking alng these lines. We wound
up with one big slab (14.6 ft x 139 ft x ~ 3 ft thick - 13 concrete trucks) and it
sits directly on un disturbed hand dug soil, and has sloped edges connecting
directly to the surrounding floors. The slopes are designed to break
transmission of low frequencies. Our FESEM picked up ~ 5.5x improvement
in resolution.

BUT we also had no major electric lines overhead or around, an
isolated electrical for the facility with lots of dedicated grounds. We used a
product called "Quite Rock" which is engineered "sheet-rock" for sound
studio walls (equivalent of 16-sheets of 3/8 inch drywall of sound
deadening). A dedicated HVAC system with laminar flows and sound
deadening in in the ducts. And a few other details.

And I think one of the biggest issues is AC electrical fields. We're
now in a "non-science" building, with a museuum, history, and anthropololgy.
By "Non-science" I mean the "hard" sciences, so no incubators, no
hundreds of computers, no ovens, and water circulators: no motors, no
transfromers, no fans, etc. All the things we lab rats usually surround
ourselves with. And all the EM Fields generated by them and their
accoustical noises are gone. Which really cleaned things up particularly with
low accelerating voltages in the SEMs. (As a test once we even shutoff the
HVAC system and still picked up ~ 30% improvement in our highest
resolution SEM.or ~ 0.9nm)

Our sites were surveyed by Integrated Dynamics Engineering (IDE), and
Peter has already spoken here. I have no fiancial ties with them, they were
good folks to work with, and we've worked with them a number of times.

The engineering specialist company we worked with was: Colin Gordon &
Associates. Again I have no fiancial ties, but they were good to work with
and gave us a very good solution.

I am asure there are similar good companies in Europe as well.

On 15 May 2013 at 4:27, dcristofori-at-unive.it wrote:

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} Dear Listers,
} we're going to install a brand new SEM with Schottky emission. We were
} thinking of position it on a base placed into a hole dug in the floor
} expressly to decouple the base from building.
}
} On the other hand we discovered that a similar base built here some 10
} years ago is not isolated that much. This base was built by digging a
} hole in the floor, than filling the bottom part of the hole with
} gravel and finally putting the concrete base on it. I think (not sure)
} the gap between the sides of the base and the hole was filled with
} sand or gravel. Finally, the same gap at floor level was filled with a
} rubber expansion joint.
}
} Do you have any better recipe? Or different solutions?
} Any comment or suggestion would be greatly appreciated!
} Thanks
}
} Davide
}
}
} ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
} Davide Cristofori
}
} direct phone = 041.234.67.26
} e-mail = dcristofori[at]unive[dot]it
}
} Università Ca' Foscari Venezia
} Dipartimento di Sc. Molecolari e Nanosistemi
} Via Torino, 155b I-30172 Mestre (VE) Italy
} ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
} "Nota automatica aggiunta dal sistema di posta
} Il 5 per mille per sostenere i giovani ricercatori di Ca' Foscari. E'
} un atto volontario, non costa nulla e non sostituisce l'8 per mille.
} Scegli Ca' Foscari: codice fiscale 80007720271 Please note that the
} above message is addressed only to individuals filing Italian income
} tax returns. -- "
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Richard E. Edelmann, Ph.D., Director
Center for Advanced Microscopy & Imaging
9C Upham Hall
Miami University, Oxford, OH 45056
Ph: 513.529.5712 Fax: 513.529.4243
E-mail: edelmare-at-miamioh.edu
http://www.cami.muohio.edu

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http://www.nytimes.com/2013/05/22/science/heinrich-rohrer-physicist-who-won=

-nobel-dies-at-79.html?ref=3Dscience

Heinrich Rohrer, who shared the 1986 Nobel Prize in

Physics for inventing a microscope that made it possible to see individual

atoms and move them around, an achievement that led to vastly faster

computing and greatly advanced molecular biology, died on Thursday

night or early

Friday morning in Wollerau, Switzerland. He was 79.

John Mardinly

ASU

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I'm making progress troubleshooting the 4 quadrant BSD on the SEM donated to our organization, but have run into a snag.

The 4QBSD is controlled by a 16 pin ribbon cable. I was extremely lucky to get full schematics from the manufacturer. It is, as I suspected, incredibly simple 5V logic. It might as well be a switch box. There are four different gain settings, and a single setting that determines detector bias and speed.

Now the software seems to be set up properly according to install instructions that a list member sent me. And I verified that SK58 on the Stage and Vacuum board (a DB25 connector) does connect to the ribbon cable.

The trouble: no matter what settings for the BSD that I enable on the SEM, I get nothing out from SK58.

By any chance, does anyone have any ideas? A pinout of SK58? Or the various passwords to get into the troubleshooting modes of the scope software?

Thanks!

Ryan

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Frank et al:
The ion pump was necessary for AMRAY to offer a LaB6 emitter, but since you are using tungsten now, the Ion pump is unnecessary.
If you just leave the column butterfly hand valve open and unplug the Ion pump HV cable all should be ok.

I'm not sure what your microscope configuration is but if you do have the butterfly valve connecting the IP to the chamber, this is a very EZ fix. The chamber vacuum gauge will read a slightly better vacuum than what is at the emission chamber but if the chamber is in the minus fives you will be good.

W filaments are cheap compared to your labor rates so even if the filament looses a small percentage of life...... Who cares....
Tungsten instruments have performed well for years with out the costly use of ion pumps and the added care they require.
If you need to keep the ion pump on line, send it to Dunniway Stock room and have it rebuilt. It's a standard Varian 30 L/sec conflat pump and they will probably have one already available for exchange.

There is no real "on instrument" fix for a old depleted ion pump. Think of them as a fancy atmospheric gas trap made of fly paper......once a pump is old and depleted every time it is started at crossover it gets warm/hot and will regurgitate some of the gasses that it has marginally trapped and buried, especially hydrogen that has been cracked from water vapor and buried under the titanium deposition. When you tap the tired pump and the pressure jumps up, what you are actually seeing is hydrogen escaping from under the loose titanium flakes.

Once you have a new pump installed try these suggestions for a quick pump down:

1) Have a spare filament gun cartridge Loaded / aligned / desiccated, and ready for insertion. Have the insides of the chamber and pump exposed to air for as short a time as possible

2) Take a heat gun and thoroughly heat the emission chamber and ion pump inside and out.
This bake out will drive out some of the H2o and significantly help the pump down time.

3) When a healthy pump is first started, the pressure will normally drop, but as the pump starts removing the residual air and water vapor it will start warming up and regurgitate some of the loosely trapped hydrogen, the pressure will start to rise, and it will need to cool off and let the turbo or diffusion pump remove some of this released hydrogen.
Just turn if off for10-20 minutes, reapply power and things should be golden. If the pump is middle aged it may require a couple of these on/off cycles but don't let the pump get too old and
Gassy

Feel free to call and discuss if this is confusing or seems outrageous......

Pete Bustanoby
SEMS
Specialized Electron Microscope Service
Serving the greater Puget Sound area since 1975

206-795-1599
cangrande65-at-yahoo.com

On May 21, 2013, at 15:17, frank_karl-at-ardl.com wrote:

}
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} Okay.
} I must nuts. We have an Amray 1803I with turbo pump and an ion getter. After we change a W filament it takes about 4-5 hours to pump down to a usable vacuum. We have to wrap on the getter, open and close valves trying to urge the system on to a good vacuum, chanting does seem to help. We define good as the alarm doesn't go off and we can work. We're not sure where the alarm position is set. Of course Amray is gone, but not forgotten
}
} I've used 5 different SEM in my 30 years as a Microscopist. I don't ever remember changing a filament taking this long, but I never had an ion getter on a SEM, only on my TEMs. Do any other Amray users have any comments or observations?
}
}
}
} Thanks............
}
} Frank
}
} This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.
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Leica Microsystems is a world leader in microscopes and scientific
instruments. Founded as a family business in the nineteenth century, the
company's history was marked by unparalleled innovation on its way to
becoming a global enterprise. It is a historically close cooperation with
the scientific community is the key to Leica Microsystems' tradition of
innovation, which draws on users' ideas and creates solutions tailored to
their requirements. At the global level, Leica Microsystems is organized
in three divisions, all of which are among the leaders in their respective
fields: the Life Science Division, Industry Division, and Medical
Division. Leica Microsystems has five manufacturing facilities in four
countries, with sales and service organizations in 20 countries. The
company is headquartered in Wetzlar, Germany Leica offers competitive
benefits including medical, dental, vision, prescription, long term care,
life insurance, STD, LTD and 401 (k). See our website at
www.leica-microsystems.com. Leica-Microsystems, Inc. is an equal
opportunity employer. M/F/D/V

Job Summary:

The Confocal Applications Specialist is a key member of the company's
sales team. The purpose of this position is to promote sales of the
companies' confocal product line in his/her assigned region. His/her
primary function is to execute domestic sales plans and to recommend
strategies to improve the sale of Confocal products in the territory, aid
in the implementation of these strategies in line with Marketing,
Corporate growth, profitability, and mission objectives.

KEY RESPONSIBILITIES:
.CUSTOMER SUPPORT: Schedules product seminars or training sessions in
concert with Leica Product & Sales Management in his/her assigned region.
Participates in all activities (travel in the field, attend industry
meetings, conferences, local and national exhibitions, etc.) that will
enhance the awareness, acceptance, and eventually the sales of Leica Life
Sciences Confocal products in his/her region.
.SATISFACTION: Insures that customers are satisfied! Also, must be
capable of working effectively with Sales Managers, PLT members, Customer
Service Technicians and Representatives, Engineers, Accounting,
Manufacturing, and Dealers to do whatever is necessary to satisfy
customers' needs.
.SALES PERFORMANCE: He/she shall conduct analysis and performance
evaluation of all sales in his/her assigned region by use of the companies
Customer Resource Management tool (CRM). The evaluation includes sales
call activities, follow up on sales leads, attendance of trade shows,
contacts with local governments, associations, and academic institutions,
and traveling with other product specialists to improve their field sales
skills and product knowledge.
.SALES PLANS: Works with Sales Management to develop strategies designed
to increase sales of Leica Confocal products in assigned territory.
Specifically, he/she recommends product, pricing, advertising and sales
promotion strategies.
.NEW PRODUCTS: He/she has responsibility to increase sales of Leica
products through the introduction of new products in his/her assigned
region. Coordinates the generation of new product ideas or modifications,
performs analysis and preliminary screening of these ideas and recommends
the appropriate ones to the Area Sales Manager for acceptance or
rejection. Works with the Marketing Department in conducting field
evaluation tests to insure the viability of the new products in his/her
assigned territory. Coordinates all activities for the introduction of
new or modified products in his/her assigned territory.
.MARKETING INFORMATION: Generates and acts on the feedback on customer
preferences, suggestions, product performance and other information to
enhance the sale of Leica Confocal products.
.APPLICATION DEVELOPMENT: Works with customers or other experts to
generate appropriate field data and application notes or publications
which highlight product benefits and helps differentiate Leica products.

Professional Experience
.Experience in working with Confocal Microscopy required
.Sales experience in selling products of a technical nature is desirable
.In-depth knowledge of the microscopy market is also highly desirable
.2 years related microscopy experience in a laboratory minimum

Education: BA, BS or MS degree in physics or chemistry or a related
scientific field

If you meet and/or exceed the experience criteria, please submit your
resume by emailing Danielle Robinson at dr-at-personifysearch.com and
reference Confocal Applications Specialist Mid-Atlantic Region LEI000739

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Dear Listers,
thank you so much to all the people giving advices and sharing their
experience. It was very useful to me to read your replies.

Just to sum up, the most of you suggested to contact an engineer
specialized in vibration isolation. On active system there is less
agreement: many stated they are the best solution, but some reported
thei experieces with systems very troublesome which failed in some year
- if not from the very beginning.

Thank you again to everybody!

Davide

~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
Davide Cristofori

direct phone = 041.234.67.26
e-mail = dcristofori[at]unive[dot]it

Universit� Ca' Foscari Venezia
Dipartimento di Sc. Molecolari e Nanosistemi
Via Torino, 155b I-30172 Mestre (VE) Italy
~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~ ~~~~~~~
"Nota automatica aggiunta dal sistema di posta
Il 5 per mille per sostenere i giovani ricercatori di Ca' Foscari.
E' un atto volontario, non costa nulla e non sostituisce l'8 per mille.
Scegli Ca' Foscari: codice fiscale 80007720271
Please note that the above message is addressed only to individuals filing
Italian income tax returns. -- "

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Email: cbc-at-queensu.ca
Name: Charlie Cooney

Organization: Queens University

Title-Subject: [Filtered] Delaminating Nicols Prism

Message: HI

I have an older model Zeiss Ultraphot with a rotating Nicol prism and it appears that prism layers
have come de-laminated. I understand that these were held together with Canada Balsam. Can I
simply re-heat the prism in the holder or is there a better way to repair this prism.

Thanks

Charlie Cooney
Queens University

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Hello Charlie,
I suspect the volatile component from the balsam has evaporated, shrinking the media and making it brittle. I would not reheat it. I'm not sure what it will do to the optical grade calcite or to what is holding the prism in place.

I suspect that unless you send it to a profession restorer you best option will be to replace the prism with a disk of polaroid film.

Why don't you join the list server and keep us informed on what happens.

Frank Karl

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Email: cbc-at-queensu.ca
Name: Charlie Cooney

Organization: Queens University

Title-Subject: [Filtered] Delaminating Nicols Prism

Message: HI

I have an older model Zeiss Ultraphot with a rotating Nicol prism and it appears that prism layers
have come de-laminated. I understand that these were held together with Canada Balsam. Can I
simply re-heat the prism in the holder or is there a better way to repair this prism.

Thanks

Charlie Cooney
Queens University

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Title-Subject: [Filtered] Zone axis pattern

Message: Can you give the explanation for relation between the reciprocal lattice vector and its
corresponding distance between the parallel lines in ZONE AXIS PATTERN

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F�r unsere Abteilung Technical Service am Standort Mannheim oder Wetzlar
suchen wir eine/n

Service-Techniker Confocal (m/w), weltweit

Verantwortungsbereiche:
� Technischer Service unserer Ger�te weltweit (Neuinstallation, Aufbau
und Reparatur)
� Betreuung wichtiger Gro�kunden
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� Ansprechpartner zu technischen Details f�r unsere
Dokumentationsgruppe
� Enge Zusammenarbeit mit der Entwicklung und Simultaneous Engineering
� �bernahme der Hotline f�r technische Fragen
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How Transfer)
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Anforderungen:
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Bereichen Elektronik, Feinmechanik und Optik, Techniker Ausbildung ist von
Vorteil)
� sehr gute Kenntnisse in Computertechnik
� sehr gute Software-Kenntnisse, F�higkeit sich schnell in neue
Software Systeme einzuarbeiten.
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� Sicheres Auftreten bei unseren Kunden und die Bereitschaft zu Reisen
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Leica Microsystems CMS GmbH ist Teil des internationalen Leica
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Microsystems zu einem erfolgreichen, weltweit agierenden Unternehmen in
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Um sich zu bewerben, E-Mail Camilla Hinton bei ch-at-personifysearch.com

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Hi Charlie,

I am not familiar with Zeiss Ultraphot, but I've used Canada Balsam for
bonding optical components and AFAIK it is not normally heated to be
liquified, but instead dissolved in Xylene. Thus I'd be afraid that just
heating the prism is not likely to work and may actually cause more
damage...

If you decide to go through the trouble of properly rebuilding your
prism then you can get liquified Canada Balsam form Sigma-Aldrich, while
the only (known to me) place where you can still purchase solid chunks
for making your own solution is

http://www.surplusshed.com/pages/item/b1077.html

Good luck :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
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Web: www.freudlabs.com

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} Title-Subject: [Filtered] Delaminating Nicols Prism
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}
} I have an older model Zeiss Ultraphot with a rotating Nicol prism and it appears that prism layers
} have come de-laminated. I understand that these were held together with Canada Balsam. Can I
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Charlie:

I'll post this on another listserve where they do light microscopy repairs
and rebuilds.

But having dealt with old objectives with balsam - do not heat it.

One can put the optical components above a saturated xylene bath overnight
and allow the xylene to soften the balsam. Then carefully press the pieces
back together (or add more xylene, pull apart, clean and reset).

Tony

.........................................
Andrew Anthony "Tony" Havics, CHMM, CIH, PE
Environmental, Health & Safety, Microscopy, & Forensic Engineering
pH2, LLC
5250 E US Highway 36, Suite 830
Avon, IN 46123
(317) 718-7020 Office
(317) 718-7038 Fax
(317) 409-3238 Cell
aahavics-at-pH2LLC.com
www.ph2LLC.com

-----Original Message-----
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Sent: Wednesday, May 29, 2013 8:09 AM
To: ph2-at-sprynet.com

Hello Charlie,
I suspect the volatile component from the balsam has evaporated, shrinking
the media and making it brittle. I would not reheat it. I'm not sure what
it will do to the optical grade calcite or to what is holding the prism in
place.

I suspect that unless you send it to a profession restorer you best option
will be to replace the prism with a disk of polaroid film.

Why don't you join the list server and keep us informed on what happens.

Frank Karl

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Email: cbc-at-queensu.ca
Name: Charlie Cooney

Organization: Queens University

Title-Subject: [Filtered] Delaminating Nicols Prism

Message: HI

I have an older model Zeiss Ultraphot with a rotating Nicol prism and it
appears that prism layers
have come de-laminated. I understand that these were held together with
Canada Balsam. Can I
simply re-heat the prism in the holder or is there a better way to repair
this prism.

Thanks

Charlie Cooney
Queens University

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Title-Subject: [Filtered] Need help processing cilia

Message: Greetings all! Does anyone have a processing schedule they would like to share, for
processing bronch washings/brushings/BAL for ciliary interpretation? In the past, all cilia studies
were done here on tissue biopsies, but we have a new clinician who prefers broncial
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Hello,

I have been trying to prepare cross-section samples of inorganic solar cells. The solar cell consists of stacked layers of material (~1-2um thick total) on a thin stainless steel (SS) substrate. The layers and the substrate together can be easily cut to different shapes using a pair of scissors. My study involves looking at the microscopic changes in these cells after they have been exposed to various temperatures for various periods of time. 50C for 30sec is the lowest temperature and time that I use.

The TEM sample preparation method that is most available to me is the polish wheel milling to dimpling to Ar-ion milling method. Before the polishing steps, the sample is prepared by gluing (Gatan G1) 2 pieces of the cells together to form a SS/cell/glue/cell/SS sandwich. The sandwich is then glued (again using G1) into a small carbon ring that fits onto the microscope's sample holder. The 3 polishing steps are then performed by mounting/demounting the sample on and off a stub using thermo wax.

In the past, I have been able to prepare these cross-section samples in this manner with no problems. For this study, however, I cannot expose the sample to even 50C for short periods of time. This causes 2 main problems. First, I cannot use thermo wax for mounting and dismounting on and off a polishing stub. Second, the G1 glue will not cure completely at room temperature, making it much less resistant to acetone and other organic solvents. I do want to note that G1 can very easily be polished and milled even when cured (48hr) at room temperature. To replace thermo wax, I have been using 2 different super glues, Loctite 460 and Pelco Samplebond, and dissolving them (during dismount) with acetone and Pelco Debonder respectively. The G1, however, did not withstand expose to acetone or the Pelco Debonder during dismounting without significant degradation. I have also tried M-bond 610 and Epofix (available to me in the lab) as replacements for G1 but they gave the same or worst results then the G1. It should be noted that samples made with G1 that was cured at 120C resulted in very satisfactory samples and were very resistant to the super glue solvents. I will soon be trying other non-EM specific epoxies.

Have others been able to prepare thin cross-section samples without exposing the sample to high temperatures (} 45C)? Can anyone suggest a room temperature curing epoxying that is resistant to organic solvents such as acetone and that also has good EM/milling properties? I also may be able to access a microtome in the future although I am just starting to learn about this technique. We have used FIB to prepare samples in the past because Ga is a key component in our samples, Ga implantation from the Ga-ion beam caused problems.

I am a graduate student and I just started learning how to use microscopy techniques for my studies. This is my first post to the list server and any advice will be greatly appreciated! Thanks in advance!

Best,

Gabriel Tong

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Hi all,

Like many other semiconductor labs, we are currently struggling with Cu surface migration
on freshly FIB'ed TEM cross sections. Of course, these samples are rush-rush.

Has anyone come up with a robust solution? I suspect part of the problem is that the Cu is like six nines pure.

One idea I had was to do a oxygen plasma chamber clean when the sample is still inside the FIB to stabilize/oxidize the Cu surfaces.

Any speculation / tips much appreciated

Bryan Tracy
Spansion, Sunnyvale

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Hi, All (or at least those who read this given the subject line)-

Since all the easy stuff has clearly already been done, lately I've been
getting the hard stuff. One trend has been people bringing in their
plant/animal/archae/cultured cells and telling me they're
dying/infected/wilting/growing tumors/behaving badly/turning pink, and
they think it may be a bacterium/virus/microsporidium/ecotoxin, and can I
look in the electron microscope, please, and see if I can find "anything".
Which is a lot of fun if they have no idea what, where, or even if.
Sampling with 60 nm sections. Yeah. Anyway, I am not a good pathologist,
and most of these issues are probably pathological.

So I've been running into a lot of veeerrry interesting intracellular and
extracellular structures. I'm pretty good at figuring out or at least
making stuff up that sounds good, but some of these things have me
stumped. I'm going to post links to some images here from time to time. If
any of you have any ideas what any of this stuff might be, speak up! Even
if you are making it up but it sounds good. I'll buy you a beer at M&M.

Maybe don't think about where the images came from, at least at first.
(Can you guess that some are from protected endangered Hawaiian tree snail
digestive gland? No? How about can you spot the coral tumor? The "it can't
possibly be contamination from my West Nile Virus"?)

OK, first set: what are these mystery intracellular tubules with a
diameter of about 53nm, bundles of which are within another membrane.
These images don't show, but they can be very long. Tubular lamellae or
lamllar tubules ... Not annulate lamellae.

https://www.dropbox.com/sc/zaomfaf8liehvsn/nc_nTtLP_V

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*
****************************************************************************

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Email: meulia.1-at-osu.edu
Name: Tea Meulia

Organization: Ohio State University

Title-Subject: [Filtered] Optical Projection Tomography scanner

Message: We wanted to purchase the Optical Projection Tomography (OPT) scanner OPT3001', from
Bioptonics, but this instrument is not in production anymore. Information on this equipment and the
type of applications that it is used for can be found at:
http://www.mrc.ac.uk/Achievementsimpact/Storiesofimpact/OpticalProjectionTomography/index.htm
http://www.bioptonics.co.uk/applications-gallery/

We have several projects that involve study on both plant organs and animal embryogenesis and we are
approaching them in the traditional way by sample embedding and sectioning. Can anyone suggest to me
another instrument similar to the Bioptonics OPT3001, that would allow scanning and 3D
reconstruction of biological specimens of ~0.5mm to 10mm.

Thank you for your help.

Tea Meulia

Director, Molecular and Cellular Imaging Center Dept.
Research Associate Professor, Dept. of Plant Pathology
Office : 330-263-3836
Lab : 330-263-3828
www.oardc.ohio-state.edu/mcic

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Sorry, people, I meant to give more information, but got sidetracked.
These are cells in (presumable healthy) digestive glands of tropical tree
snails. I am looking at thses as "normal" tissues to compare with the
putrified digestive glands of endangered native Hawaiian tree snails that
are dying, and we can't kill any living ones to see what they *should*
look like. So think about digestion, not photoreception or photosynthesis
(although lately I've seen some improbable things).

I have more mysteries I hope to post tomorrow. Crowdsourcing cell biology?

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*
****************************************************************************

==============================Original Headers==============================
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So, Tina, what were you referring to as pathological, the samples or those
who bring the samples?
Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

-----Original Message-----
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Sent: Thursday, May 30, 2013 4:58 PM
To: kenconverse-at-qualityimages.biz

Hi, All (or at least those who read this given the subject line)-

Since all the easy stuff has clearly already been done, lately I've been
getting the hard stuff. One trend has been people bringing in their
plant/animal/archae/cultured cells and telling me they're
dying/infected/wilting/growing tumors/behaving badly/turning pink, and
they think it may be a bacterium/virus/microsporidium/ecotoxin, and can I
look in the electron microscope, please, and see if I can find "anything".
Which is a lot of fun if they have no idea what, where, or even if.
Sampling with 60 nm sections. Yeah. Anyway, I am not a good pathologist,
and most of these issues are probably pathological.

So I've been running into a lot of veeerrry interesting intracellular and
extracellular structures. I'm pretty good at figuring out or at least
making stuff up that sounds good, but some of these things have me
stumped. I'm going to post links to some images here from time to time. If
any of you have any ideas what any of this stuff might be, speak up! Even
if you are making it up but it sounds good. I'll buy you a beer at M&M.

Maybe don't think about where the images came from, at least at first.
(Can you guess that some are from protected endangered Hawaiian tree snail
digestive gland? No? How about can you spot the coral tumor? The "it can't
possibly be contamination from my West Nile Virus"?)

OK, first set: what are these mystery intracellular tubules with a
diameter of about 53nm, bundles of which are within another membrane.
These images don't show, but they can be very long. Tubular lamellae or
lamllar tubules ... Not annulate lamellae.

https://www.dropbox.com/sc/zaomfaf8liehvsn/nc_nTtLP_V

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*
****************************************************************************

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Tina,

Enigmasomes, obviously.

Or:
They look very much like the tubular ER in the Arion rufus (Pulmonata)
digestive gland.
Another thought: are these endothelial cells in the digestive gland?
Could be microtubules associated with microvilli?

Phil

} Hi, All (or at least those who read this given the subject line)-
}
} Since all the easy stuff has clearly already been done, lately I've been
} getting the hard stuff. One trend has been people bringing in their
} plant/animal/archae/cultured cells and telling me they're
} dying/infected/wilting/growing tumors/behaving badly/turning pink, and
} they think it may be a bacterium/virus/microsporidium/ecotoxin, and can I
} look in the electron microscope, please, and see if I can find "anything".
} Which is a lot of fun if they have no idea what, where, or even if.
} Sampling with 60 nm sections. Yeah. Anyway, I am not a good pathologist,
} and most of these issues are probably pathological.
}
} So I've been running into a lot of veeerrry interesting intracellular and
} extracellular structures. I'm pretty good at figuring out or at least
} making stuff up that sounds good, but some of these things have me
} stumped. I'm going to post links to some images here from time to time. If
} any of you have any ideas what any of this stuff might be, speak up! Even
} if you are making it up but it sounds good. I'll buy you a beer at M&M.
}
} Maybe don't think about where the images came from, at least at first.
} (Can you guess that some are from protected endangered Hawaiian tree snail
} digestive gland? No? How about can you spot the coral tumor? The "it can't
} possibly be contamination from my West Nile Virus"?)
}
} OK, first set: what are these mystery intracellular tubules with a
} diameter of about 53nm, bundles of which are within another membrane.
} These images don't show, but they can be very long. Tubular lamellae or
} lamllar tubules ... Not annulate lamellae.
}
} https://www.dropbox.com/sc/zaomfaf8liehvsn/nc_nTtLP_V
}
} 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*
} ****************************************************************************

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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If you look closely, the clusters of tubules seem to be surrounded by a membrane - so I don't think it is something simple like oblique sections of microvilli or cilia

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, May 31, 2013 8:49 AM
To: Phillips, Thomas E.

Tina,

Enigmasomes, obviously.

Or:
They look very much like the tubular ER in the Arion rufus (Pulmonata) digestive gland.
Another thought: are these endothelial cells in the digestive gland?
Could be microtubules associated with microvilli?

Phil

} Hi, All (or at least those who read this given the subject line)-
}
} Since all the easy stuff has clearly already been done, lately I've
} been getting the hard stuff. One trend has been people bringing in
} their plant/animal/archae/cultured cells and telling me they're
} dying/infected/wilting/growing tumors/behaving badly/turning pink, and
} they think it may be a bacterium/virus/microsporidium/ecotoxin, and
} can I look in the electron microscope, please, and see if I can find "anything".
} Which is a lot of fun if they have no idea what, where, or even if.
} Sampling with 60 nm sections. Yeah. Anyway, I am not a good
} pathologist, and most of these issues are probably pathological.
}
} So I've been running into a lot of veeerrry interesting intracellular
} and extracellular structures. I'm pretty good at figuring out or at
} least making stuff up that sounds good, but some of these things have
} me stumped. I'm going to post links to some images here from time to
} time. If any of you have any ideas what any of this stuff might be,
} speak up! Even if you are making it up but it sounds good. I'll buy you a beer at M&M.
}
} Maybe don't think about where the images came from, at least at first.
} (Can you guess that some are from protected endangered Hawaiian tree
} snail digestive gland? No? How about can you spot the coral tumor? The
} "it can't possibly be contamination from my West Nile Virus"?)
}
} OK, first set: what are these mystery intracellular tubules with a
} diameter of about 53nm, bundles of which are within another membrane.
} These images don't show, but they can be very long. Tubular lamellae
} or lamllar tubules ... Not annulate lamellae.
}
} https://www.dropbox.com/sc/zaomfaf8liehvsn/nc_nTtLP_V
}
} 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*
} **********************************************************************
} ******

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Bryan,
I don't know much about Cu migration across the surface of a FIB prepared
sample, but I'm interested in hearing more about it.

I was experimenting with sample preparation of copper for EBSD analysis. I
can tell you that I took a polished, OFHC copper sample, ion polished it
with low angle, low energy Ar ions, and then plasma cleaned it with 100% O2,
5 W for 5 minutes and you could see that it had oxidized the surface. You
will need to control that process to limit the thickness of any oxide layer
that you grow on the sample.

-Scott

-----Original Message-----
X-from: Bryan.Tracy-at-spansion.com [mailto:Bryan.Tracy-at-spansion.com]
Sent: Thursday, May 30, 2013 1:21 PM
To: S.Walck-at-comcast.net

Hi all,

Like many other semiconductor labs, we are currently struggling with Cu
surface migration on freshly FIB'ed TEM cross sections. Of course, these
samples are rush-rush.

Has anyone come up with a robust solution? I suspect part of the problem is
that the Cu is like six nines pure.

One idea I had was to do a oxygen plasma chamber clean when the sample is
still inside the FIB to stabilize/oxidize the Cu surfaces.

Any speculation / tips much appreciated

Bryan Tracy
Spansion, Sunnyvale

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Hi Tina

Assuming the snail digestive gland; Snail endothelial ciliary basal bodies. These could be sections through the basal corpuscle of a ciliated cell. The tubules resemble the fibrillar rootlet structures found in the basal corpuscle just below the cell surface within the apical cytoplasm of other organisms. Although the lack of any familiar 9x2 cilia structure and the presence of a surrounding membrane, as Thomas noted, suggest otherwise, this may suggest the structures are microvilli. The membrane sheath could be related to a lateral knob structure or rootlet structure seen around microtubules in cilia, or micropinocytotic vesicle-related membrane invaginations (admittedly unlikely).
Do the surfaces of these cells show any microvilli/cilia structures? Maybe a primitive form of invertebrate cilia? 3 cytoplasm shots may not be enough to judge, although image 3 looks like it is near the surface.

Chris Santeufemio
TEM-FIB-SEM Principal Scientist
U. Mass Lowell
978-934-2429

________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Friday, May 31, 2013 10:06 AM
To: Santeufemio, Christopher

If you look closely, the clusters of tubules seem to be surrounded by a membrane - so I don't think it is something simple like oblique sections of microvilli or cilia

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, May 31, 2013 8:49 AM
To: Phillips, Thomas E.

Tina,

Enigmasomes, obviously.

Or:
They look very much like the tubular ER in the Arion rufus (Pulmonata) digestive gland.
Another thought: are these endothelial cells in the digestive gland?
Could be microtubules associated with microvilli?

Phil

} Hi, All (or at least those who read this given the subject line)-
}
} Since all the easy stuff has clearly already been done, lately I've
} been getting the hard stuff. One trend has been people bringing in
} their plant/animal/archae/cultured cells and telling me they're
} dying/infected/wilting/growing tumors/behaving badly/turning pink, and
} they think it may be a bacterium/virus/microsporidium/ecotoxin, and
} can I look in the electron microscope, please, and see if I can find "anything".
} Which is a lot of fun if they have no idea what, where, or even if.
} Sampling with 60 nm sections. Yeah. Anyway, I am not a good
} pathologist, and most of these issues are probably pathological.
}
} So I've been running into a lot of veeerrry interesting intracellular
} and extracellular structures. I'm pretty good at figuring out or at
} least making stuff up that sounds good, but some of these things have
} me stumped. I'm going to post links to some images here from time to
} time. If any of you have any ideas what any of this stuff might be,
} speak up! Even if you are making it up but it sounds good. I'll buy you a beer at M&M.
}
} Maybe don't think about where the images came from, at least at first.
} (Can you guess that some are from protected endangered Hawaiian tree
} snail digestive gland? No? How about can you spot the coral tumor? The
} "it can't possibly be contamination from my West Nile Virus"?)
}
} OK, first set: what are these mystery intracellular tubules with a
} diameter of about 53nm, bundles of which are within another membrane.
} These images don't show, but they can be very long. Tubular lamellae
} or lamllar tubules ... Not annulate lamellae.
}
} https://www.dropbox.com/sc/zaomfaf8liehvsn/nc_nTtLP_V
}
} 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*
} **********************************************************************
} ******

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

I don't follow the logic of these being microtubules inside cilia - not only are they not 9+2 or even 9+0 (as one would find in non-motile primary cilia, the number of circular structures varies per cluster. Microtubules in the outer ring of either the 9+0 or 9+2 cores of cilia are doublets that look like a figure 8 and there is no hint of that here. Finally, my eyeball estimate suggests they they are too wide for microtubules (25 nm).

The open endedness of the structures in Figure 2 made me think of Birbeck granules but I doubt they are present in invertebrates.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: Christopher_Santeufemio-at-uml.edu [mailto:Christopher_Santeufemio-at-uml.edu]
Sent: Friday, May 31, 2013 11:05 AM
To: Phillips, Thomas E.

Hi Tina

Assuming the snail digestive gland; Snail endothelial ciliary basal bodies. These could be sections through the basal corpuscle of a ciliated cell. The tubules resemble the fibrillar rootlet structures found in the basal corpuscle just below the cell surface within the apical cytoplasm of other organisms. Although the lack of any familiar 9x2 cilia structure and the presence of a surrounding membrane, as Thomas noted, suggest otherwise, this may suggest the structures are microvilli. The membrane sheath could be related to a lateral knob structure or rootlet structure seen around microtubules in cilia, or micropinocytotic vesicle-related membrane invaginations (admittedly unlikely).
Do the surfaces of these cells show any microvilli/cilia structures? Maybe a primitive form of invertebrate cilia? 3 cytoplasm shots may not be enough to judge, although image 3 looks like it is near the surface.

Chris Santeufemio
TEM-FIB-SEM Principal Scientist
U. Mass Lowell
978-934-2429

________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Friday, May 31, 2013 10:06 AM
To: Santeufemio, Christopher

If you look closely, the clusters of tubules seem to be surrounded by a membrane - so I don't think it is something simple like oblique sections of microvilli or cilia

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, May 31, 2013 8:49 AM
To: Phillips, Thomas E.

Tina,

Enigmasomes, obviously.

Or:
They look very much like the tubular ER in the Arion rufus (Pulmonata) digestive gland.
Another thought: are these endothelial cells in the digestive gland?
Could be microtubules associated with microvilli?

Phil

} Hi, All (or at least those who read this given the subject line)-
}
} Since all the easy stuff has clearly already been done, lately I've
} been getting the hard stuff. One trend has been people bringing in
} their plant/animal/archae/cultured cells and telling me they're
} dying/infected/wilting/growing tumors/behaving badly/turning pink, and
} they think it may be a bacterium/virus/microsporidium/ecotoxin, and
} can I look in the electron microscope, please, and see if I can find "anything".
} Which is a lot of fun if they have no idea what, where, or even if.
} Sampling with 60 nm sections. Yeah. Anyway, I am not a good
} pathologist, and most of these issues are probably pathological.
}
} So I've been running into a lot of veeerrry interesting intracellular
} and extracellular structures. I'm pretty good at figuring out or at
} least making stuff up that sounds good, but some of these things have
} me stumped. I'm going to post links to some images here from time to
} time. If any of you have any ideas what any of this stuff might be,
} speak up! Even if you are making it up but it sounds good. I'll buy you a beer at M&M.
}
} Maybe don't think about where the images came from, at least at first.
} (Can you guess that some are from protected endangered Hawaiian tree
} snail digestive gland? No? How about can you spot the coral tumor? The
} "it can't possibly be contamination from my West Nile Virus"?)
}
} OK, first set: what are these mystery intracellular tubules with a
} diameter of about 53nm, bundles of which are within another membrane.
} These images don't show, but they can be very long. Tubular lamellae
} or lamllar tubules ... Not annulate lamellae.
}
} https://www.dropbox.com/sc/zaomfaf8liehvsn/nc_nTtLP_V
}
} 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*
} **********************************************************************
} ******

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Hi,

We have a Philips 400T which we want to dispose. After many productive years, in 2008 a fuse was blowing constantly and as the use was reduced we switched it off.

In addition, we have side equipment to this microscope, which include a dismantled 400T which we used for spare part.

Please contact offline if you are interested in these columns.

Kind regards,

Ton van Helvoort

Antonius (Ton) T.J. van Helvoort
Associate professor
Department of Physics
Norwegian University of Science and Technology (NTNU)
NO-7491 Trondheim, Norway
Location: Realfagsbygd, room D4-149
Tel: (0047)-735-93637
email: a.helvoort-at-ntnu.no
http://folk.ntnu.no/helvoort/
http://www.ntnu.edu/web/geminicentre/tem

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Hi listers,
I wonder if I have a really bad case of static (please, see TEM picture in dropbox). It is just section of embedding resin, no specimen in this field of view.
Any advice on how to get rid of it?
Thanks,
Vladimir
https://www.dropbox.com/sh/ddm8z2m4bhfxcqu/Ul4ZIZ4WzO

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

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I love a puzzle, so this got me looking for an answer as I've never seen tubules like this before. A google search of the simple terms
snail organelles electron
brought up this article
Abolins-Krogis A. Alterations in the fine structure of cytoplasmic organelles in the hepatopancreatic cells of shell-regenerating snail, Helix pomatia L. Z Zellforsch Mikrosk Anat. 1970;108(4):516-29.

http://link.springer.com/article/10.1007%2FBF00339657
The article has a few images that are similar to the ones you show (particularly fig 3: Activated digestive cell).

The suggestion seems to be that the tubules in the midgut are involved in repair to the shell. The author seems to have a few papers looking at these sort of organelles, but I feel that I've already read too much for today.

This may well be irrelevant, but it looks similar and is from a similar species, so hopefully it is useful. It was certainly an interesting puzzle.

Toby S

Dr Tobias Starborg
Senior Experimental Officer: 3DEM
Wellcome Centre for Cell Matrix Research
Michael Smith Building
Manchester
M13 9PT
Tel: +44(0)1612755170
http://manchesterpolara.org.uk

________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: 31 May 2013 17:34
To: Tobias Starborg

I don't follow the logic of these being microtubules inside cilia - not only are they not 9+2 or even 9+0 (as one would find in non-motile primary cilia, the number of circular structures varies per cluster. Microtubules in the outer ring of either the 9+0 or 9+2 cores of cilia are doublets that look like a figure 8 and there is no hint of that here. Finally, my eyeball estimate suggests they they are too wide for microtubules (25 nm).

The open endedness of the structures in Figure 2 made me think of Birbeck granules but I doubt they are present in invertebrates.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: Christopher_Santeufemio-at-uml.edu [mailto:Christopher_Santeufemio-at-uml.edu]
Sent: Friday, May 31, 2013 11:05 AM
To: Phillips, Thomas E.

Hi Tina

Assuming the snail digestive gland; Snail endothelial ciliary basal bodies. These could be sections through the basal corpuscle of a ciliated cell. The tubules resemble the fibrillar rootlet structures found in the basal corpuscle just below the cell surface within the apical cytoplasm of other organisms. Although the lack of any familiar 9x2 cilia structure and the presence of a surrounding membrane, as Thomas noted, suggest otherwise, this may suggest the structures are microvilli. The membrane sheath could be related to a lateral knob structure or rootlet structure seen around microtubules in cilia, or micropinocytotic vesicle-related membrane invaginations (admittedly unlikely).
Do the surfaces of these cells show any microvilli/cilia structures? Maybe a primitive form of invertebrate cilia? 3 cytoplasm shots may not be enough to judge, although image 3 looks like it is near the surface.

Chris Santeufemio
TEM-FIB-SEM Principal Scientist
U. Mass Lowell
978-934-2429

________________________________________
X-from: PhillipsT-at-missouri.edu [PhillipsT-at-missouri.edu]
Sent: Friday, May 31, 2013 10:06 AM
To: Santeufemio, Christopher

If you look closely, the clusters of tubules seem to be surrounded by a membrane - so I don't think it is something simple like oblique sections of microvilli or cilia

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, May 31, 2013 8:49 AM
To: Phillips, Thomas E.

Tina,

Enigmasomes, obviously.

Or:
They look very much like the tubular ER in the Arion rufus (Pulmonata) digestive gland.
Another thought: are these endothelial cells in the digestive gland?
Could be microtubules associated with microvilli?

Phil

} Hi, All (or at least those who read this given the subject line)-
}
} Since all the easy stuff has clearly already been done, lately I've
} been getting the hard stuff. One trend has been people bringing in
} their plant/animal/archae/cultured cells and telling me they're
} dying/infected/wilting/growing tumors/behaving badly/turning pink, and
} they think it may be a bacterium/virus/microsporidium/ecotoxin, and
} can I look in the electron microscope, please, and see if I can find "anything".
} Which is a lot of fun if they have no idea what, where, or even if.
} Sampling with 60 nm sections. Yeah. Anyway, I am not a good
} pathologist, and most of these issues are probably pathological.
}
} So I've been running into a lot of veeerrry interesting intracellular
} and extracellular structures. I'm pretty good at figuring out or at
} least making stuff up that sounds good, but some of these things have
} me stumped. I'm going to post links to some images here from time to
} time. If any of you have any ideas what any of this stuff might be,
} speak up! Even if you are making it up but it sounds good. I'll buy you a beer at M&M.
}
} Maybe don't think about where the images came from, at least at first.
} (Can you guess that some are from protected endangered Hawaiian tree
} snail digestive gland? No? How about can you spot the coral tumor? The
} "it can't possibly be contamination from my West Nile Virus"?)
}
} OK, first set: what are these mystery intracellular tubules with a
} diameter of about 53nm, bundles of which are within another membrane.
} These images don't show, but they can be very long. Tubular lamellae
} or lamllar tubules ... Not annulate lamellae.
}
} https://www.dropbox.com/sc/zaomfaf8liehvsn/nc_nTtLP_V
}
} 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*
} **********************************************************************
} ******

--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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It was digital picture. Still a lot nearly parallel "lightening crackles"...
Embedding resin was EMbed812.

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: Schwarz, Janet E [mailto:janet.schwarz-at-med.uvm.edu]
Sent: Friday, May 31, 2013 1:46 PM
To: Dusevich, Vladimir

Hi listers,
I wonder if I have a really bad case of static (please, see TEM picture in dropbox). It is just section of embedding resin, no specimen in this field of view.
Any advice on how to get rid of it?
Thanks,
Vladimir
https://www.dropbox.com/sh/ddm8z2m4bhfxcqu/Ul4ZIZ4WzO

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

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Email: flaitz-at-us.ibm.com
Name: Philip Flaitz

Organization: IBM Microelectronics

Title-Subject: [Filtered] Re: [Microscopy] Cu surface migration on freshly FIB'ed Cu seed TEM cross
sections

Message:
Hi, Bryan. Isn't Cu fun?!? :-(

I will give you a few factors we have observed and implemented in our lab.

1. FIB prep. is *way* better than polishing and ion milling. It can be done, but you have extreme
challenges. With FIB prep., you cannot do low KV final thinning, which we have started doing to
minimize amorphization of the Si. We have found our cutoff to be ~8KV; if we use that, we have not
observed extensive Cu corrosion problems. The Ga beam appears to induce a small amount of surface
mixing which makes the structure more robust to corrosion, and more so at higher KV. This probably
also explains why polishing plus Ar milling has so many problems; the KV is generally quite low
( {3KV) and the surfaces are too pristine to avoid corrosion.

2. The samples should be stored under vacuum, with a nitrogen purge when venting. This keeps
moisture away from the sample, and moisture is the key problem. (Plus whatever contaminants your
local power generating station is pumping into the air!!) We tried a flowing dry nitrogen cabinet,
but vacuum has worked much better, probably because the pumpdown removes the air around the sample,
and the next purge surrounds it with nitrogen.

3. Do *not* plasma clean unless absolutely necessary. With vacuum storage, we generally find that
it is not needed. Some samples, however, do have severe contamination, usually from a dirty grid.
These will have to be plasma cleaned for STEM or analysis, but you will likely find corrosion of the
Cu if you go back to the sample again.

4. If you have used Enhanced Etch (iodine) in your FIB, you are really stuck. The iodine will end
up accumulating on the interior surfaces, and will spontaneously react with Cu to form CuI2. A
chamber clean plus a bake might clear things up if the GIS has been removed.

Let me know if any of this helps. Since we implemented vacuum storage and } 8KV FIB beam, we have
had virtually no problem with Cu corrosion, and have been able to revisit stored samples for up to a
few weeks.

regards,
Phil

Philip L. Flaitz
IBM Microelectronics, Hopewell Junction, NY
Ph.......(845) 892-3094, FAX -6256
pager - 800-608-9398
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}
}
} Bryan;
} When Intel first started copper, the TEM labs were borrowing time on FIBs that were used for F/A, all with iodine or xenon difluoride. The samples that came out of these FIBs tended to be covered with "fur" by the time they got to the FIB. What we found was that whenever a FIB had ever been used for enhanced etch, it was never again suitable for use as a TEM prep tool for copper. This required purchasing new FIBs dedicated to TEM prep, something that did not go down well with management! However, it did solve the problem. The other approach that helped a lot was vacuum storage of the specimens. We purchased a Gatan unit, specially designed for specimen storage. It used the vacuum system of the model 655 specimen holder. You can see the design on their web page. This worked well for some time. However, a word of caution: You MUST change the rubber diaphragms annually, without fail. If the diaphragms develop cracks, it will back stream sulphur containing motor oil into the system, contaminating everything and corroding the copper. Our $17,000 system had to be scrapped because the molecular drag pump was contaminated, and it could not be successfully cleaned.
} My experience with the nightmare of copper returned after Intel closed in Santa Clara, and I moved to Western Digital Media R&D. Some experimental disks contained a layer of copper. Before we had our own FIB, we had all of our samples prepared by a local vendor of TEM services. On a few occasions, their weekend shift used an 855 (with enhanced etch installed but not used) just to do the lift out and glue the 1 micron lamellae to the copper grids. They were then finished in a new Helios. By the time they got to the TEM, they were destroyed. I knew what the problem was right away, but nobody believed me. Eventually, they came to accept the idea that copper could never go into a FIB with enhanced etch installed, whether it was being used or not. Unmeasurable traces of that stuff just destroy copper TEM samples.
}
} John Mardinly, ASU
}
} On May 30, 2013, at 10:18 AM, Bryan.Tracy-at-spansion.com wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
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} }
} } Hi all,
} }
} } Like many other semiconductor labs, we are currently struggling with Cu surface migration
} } on freshly FIB'ed TEM cross sections. Of course, these samples are rush-rush.
} }
} } Has anyone come up with a robust solution? I suspect part of the problem is that the Cu is like six nines pure.
} }
} } One idea I had was to do a oxygen plasma chamber clean when the sample is still inside the FIB to stabilize/oxidize the Cu surfaces.
} }
} } Any speculation / tips much appreciated
} }
} }
} } Bryan Tracy
} } Spansion, Sunnyvale
} }
} }
} } ==============================Original Headers==============================
} } 8, 27 -- From Bryan.Tracy-at-spansion.com Thu May 30 12:08:38 2013
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} } 8, 27 -- Subject: Cu surface migration on freshly FIB'ed Cu seed TEM cross sections
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3, 39 -- Subject: Fwd: [Microscopy] Cu surface migration on freshly FIB'ed Cu seed
3, 39 -- TEM cross sections
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3, 39 -- TEM cross sections
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Bryan;
One other thought came to mind a few hours after I first replied, and that was how we dealt with wedge or dimpled and ion milled samples. As I recall, and it has been a few years, we typically had corrosion problems if the sample was not ion milled immediately after dimple or wedge polish. This idea popped up after one technician polished a batch of samples on a Friday, cleaned and dried them, but then when they were milled on Monday, they all corroded instantly after milling. One thought was that something in the Syton was diffusing into the samples, perhaps into the epoxy. One suggestion was that it was the phosphoric acid in the Syton. In any event, when we adopted the procedure that ion milling was done immediately after polishing, the problem of copper corrosion on argon milled samples faded into the background�.until of course, the rubber diaphragms on our vacuum storage system cracked. Remember, miniscule amounts of the wrong materials seem to be able to have a catalytic effect on the corrosion rate of copper.

John Mardinly, ASU

On Jun 2, 2013, at 12:44 PM, John Mardinly {John.Mardinly-at-asu.edu} wrote:

} }
} }
} } Bryan;
} } When Intel first started copper, the TEM labs were borrowing time on FIBs that were used for F/A, all with iodine or xenon difluoride. The samples that came out of these FIBs tended to be covered with "fur" by the time they got to the FIB. What we found was that whenever a FIB had ever been used for enhanced etch, it was never again suitable for use as a TEM prep tool for copper. This required purchasing new FIBs dedicated to TEM prep, something that did not go down well with management! However, it did solve the problem. The other approach that helped a lot was vacuum storage of the specimens. We purchased a Gatan unit, specially designed for specimen storage. It used the vacuum system of the model 655 specimen holder. You can see the design on their web page. This worked well for some time. However, a word of caution: You MUST change the rubber diaphragms annually, without fail. If the diaphragms develop cracks, it will back stream sulphur containing motor oil into the system, contaminating everything and corroding the copper. Our $17,000 system had to be scrapped because the molecular drag pump was contaminated, and it could not be successfully cleaned.
} } My experience with the nightmare of copper returned after Intel closed in Santa Clara, and I moved to Western Digital Media R&D. Some experimental disks contained a layer of copper. Before we had our own FIB, we had all of our samples prepared by a local vendor of TEM services. On a few occasions, their weekend shift used an 855 (with enhanced etch installed but not used) just to do the lift out and glue the 1 micron lamellae to the copper grids. They were then finished in a new Helios. By the time they got to the TEM, they were destroyed. I knew what the problem was right away, but nobody believed me. Eventually, they came to accept the idea that copper could never go into a FIB with enhanced etch installed, whether it was being used or not. Unmeasurable traces of that stuff just destroy copper TEM samples.
} }
} } John Mardinly, ASU
} }
} } On May 30, 2013, at 10:18 AM, Bryan.Tracy-at-spansion.com wrote:
} }
} } }
} } }
} } }
} } } ----------------------------------------------------------------------------
} } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } }
} } } Hi all,
} } }
} } } Like many other semiconductor labs, we are currently struggling with Cu surface migration
} } } on freshly FIB'ed TEM cross sections. Of course, these samples are rush-rush.
} } }
} } } Has anyone come up with a robust solution? I suspect part of the problem is that the Cu is like six nines pure.
} } }
} } } One idea I had was to do a oxygen plasma chamber clean when the sample is still inside the FIB to stabilize/oxidize the Cu surfaces.
} } }
} } } Any speculation / tips much appreciated
} } }
} } }
} } } Bryan Tracy
} } } Spansion, Sunnyvale
} } }
} } }
} } } ==============================Original Headers==============================
} } } 8, 27 -- From Bryan.Tracy-at-spansion.com Thu May 30 12:08:38 2013
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} } } 8, 27 -- To: "Listserver (Microscopy-at-microscopy.com)" {Microscopy-at-microscopy.com}
} } } 8, 27 -- CC: Vince Carlino {vince.carlino-at-ibssgroup.com}
} } } 8, 27 -- Subject: Cu surface migration on freshly FIB'ed Cu seed TEM cross sections
} } } 8, 27 -- Thread-Topic: Cu surface migration on freshly FIB'ed Cu seed TEM cross
} } } 8, 27 -- sections
} } } 8, 27 -- Thread-Index: Ac5dWFIZUqIfE31ASYWThOKPUcqe7w==
} } } 8, 27 -- Date: Thu, 30 May 2013 17:08:36 +0000
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7, 43 -- To: MSA Listserver {Microscopy-at-Microscopy.com} ,
7, 43 -- Bryan Tracy
7, 43 -- {Bryan.Tracy-at-spansion.com}
7, 43 -- Subject: Re: [Microscopy] Cu surface migration on freshly FIB'ed Cu seed TEM
7, 43 -- cross sections
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7, 43 -- TEM cross sections
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} Ian;
} As I mentioned, Intel had to purchase at least one dedicated small chamber FIB at each of their sites that did TEM, and let me tell you, that did not go over well because Intel is extremely cost conscious about lab equipment. Of course FEI knew about it-they were selling us the FIBs! However, FEI is a big company, and not everyone knows everything. BTW, I am not familiar with the abbreviation IEE. Our experience was with iodine and xenon difluoride, and we never found any success with wiping down the walls of the big FIBs. Besides, the yield and F/A guys still had an ongoing need for enhanced etch, so we had to have dedicated clean systems for doing copper. As for images, sorry, we tended to not waste time taking pictures of ruined samples.
}
} --John Mardinly, ASU
}
}
} On Jun 2, 2013, at 1:33 PM, Ian Drucker {ian.drucker-at-gmail.com} wrote:
}
} } Hello,
} }
} } I saw your reply on the List Server regarding Copper problems in FIB's with IEE. I work for Applied Materials and we use a Helios600i to do cross sections of copper samples. Either TSV's or Damascene trenches most of the time. We're going to be doing so STEM work in the near future for the first time and your response has worried me. We have IEE on our Helios and were never told by FEI about this problem. I'm not sure I quite understand what your seeing though when you have a problem. Also if we remove the IEE and clean the chamber walls and run some plasma cleans would that reduce and or eliminate the problem? Do you have any images of the issues you saw in the past?
} }
} } Thanks
} }
} } Ian_Drucker-at-amat.com
} }
} }
} }
} }
} } On Sun, Jun 2, 2013 at 1:55 PM, {John.Mardinly-at-asu.edu} wrote:
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
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} }
} } }
} } }
} } } Bryan;
} } } When Intel first started copper, the TEM labs were borrowing time on FIBs that were used for F/A, all with iodine or xenon difluoride. The samples that came out of these FIBs tended to be covered with "fur" by the time they got to the FIB. What we found was that whenever a FIB had ever been used for enhanced etch, it was never again suitable for use as a TEM prep tool for copper. This required purchasing new FIBs dedicated to TEM prep, something that did not go down well with management! However, it did solve the problem. The other approach that helped a lot was vacuum storage of the specimens. We purchased a Gatan unit, specially designed for specimen storage. It used the vacuum system of the model 655 specimen holder. You can see the design on their web page. This worked well for some time. However, a word of caution: You MUST change the rubber diaphragms annually, without fail. If the diaphragms develop cracks, it will back stream sulphur containing motor oil into th!
} } e system, contaminating everything and corroding the copper. Our $17,000 system had to be scrapped because the molecular drag pump was contaminated, and it could not be successfully cleaned.
} } } My experience with the nightmare of copper returned after Intel closed in Santa Clara, and I moved to Western Digital Media R&D. Some experimental disks contained a layer of copper. Before we had our own FIB, we had all of our samples prepared by a local vendor of TEM services. On a few occasions, their weekend shift used an 855 (with enhanced etch installed but not used) just to do the lift out and glue the 1 micron lamellae to the copper grids. They were then finished in a new Helios. By the time they got to the TEM, they were destroyed. I knew what the problem was right away, but nobody believed me. Eventually, they came to accept the idea that copper could never go into a FIB with enhanced etch installed, whether it was being used or not. Unmeasurable traces of that stuff just destroy copper TEM samples.
} } }
} } } John Mardinly, ASU
} } }
} } } On May 30, 2013, at 10:18 AM, Bryan.Tracy-at-spansion.com wrote:
} } }
} } } }
} } } }
} } } }
} } } } ----------------------------------------------------------------------------
} } } } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } } } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } } } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } } } ----------------------------------------------------------------------------
} } } }
} } } } Hi all,
} } } }
} } } } Like many other semiconductor labs, we are currently struggling with Cu surface migration
} } } } on freshly FIB'ed TEM cross sections. Of course, these samples are rush-rush.
} } } }
} } } } Has anyone come up with a robust solution? I suspect part of the problem is that the Cu is like six nines pure.
} } } }
} } } } One idea I had was to do a oxygen plasma chamber clean when the sample is still inside the FIB to stabilize/oxidize the Cu surfaces.
} } } }
} } } } Any speculation / tips much appreciated
} } } }
} } } }
} } } } Bryan Tracy
} } } } Spansion, Sunnyvale
} } } }
} } } }
} } } } ==============================Original Headers==============================
} } } } 8, 27 -- From Bryan.Tracy-at-spansion.com Thu May 30 12:08:38 2013
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} } } } 8, 27 -- To: "Listserver (Microscopy-at-microscopy.com)" {Microscopy-at-microscopy.com}
} } } } 8, 27 -- CC: Vince Carlino {vince.carlino-at-ibssgroup.com}
} } } } 8, 27 -- Subject: Cu surface migration on freshly FIB'ed Cu seed TEM cross sections
} } } } 8, 27 -- Thread-Topic: Cu surface migration on freshly FIB'ed Cu seed TEM cross
} } } } 8, 27 -- sections
} } } } 8, 27 -- Thread-Index: Ac5dWFIZUqIfE31ASYWThOKPUcqe7w==
} } } } 8, 27 -- Date: Thu, 30 May 2013 17:08:36 +0000
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} } 3, 39 -- To: MSA Listserver {Microscopy-at-Microscopy.com}
} } 3, 39 -- Subject: Fwd: [Microscopy] Cu surface migration on freshly FIB'ed Cu seed
} } 3, 39 -- TEM cross sections
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} } 3, 39 -- TEM cross sections
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Thank you all for your participation! I wish it was a contest and I could
award a prize if anyone figures it out.

These are appearing in both pathological (and very dead) and healthy
tissues.

They are absolutely not microtubules, any part of cilia, flagellae or
centrosomes, or microvilli. Trust me on this; I've worked for decades on
various ciliary structures in invertebrates. It's animal (tree snail), so
not any plant parts. (OK, I know there are symbiotic algae in some snails,
and I've actually worked on those, but this is not that.) They do not seem
to be rhabdo- or-any-other-virus, although I will not absolutely rule that
out.

They do look most similar to membranes reported in some types of muscle,
and while these cells are not where I expect muscle to be, I'll look again
since they could be part of the tubule system in muscle around the
digestive gland (and I've looked at copepod muscle that has a T-tubule
system that runs perpendicular to everyone else's and looks exotic),
thanks Omar Skalli. Tobias Baskin led me to some papers from the early
1970s that show similar arrays of smooth ER in snails that are undergoing
shell repair, among other things. I think that might be the ticket, if
only I could lay my hands on the papers (grrr). I hope to have more time
to look at these structures later this week.

Stay tuned for more mystery structures!

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*
****************************************************************************

==============================Original Headers==============================
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Title-Subject: [Filtered] What to do with film plates and cassettes?

Message: When I got my monochromated F20 in 2003 it came with 200 plates and 4 cassettes. I kept
them....just in case. Well, just in case never happened and now I have all this brand-stinking new
and useless scope stuff cluttering up the room. If anyone wants them, god bless you, give me your
fedx number and enjoy. In lieu of that, they're not pretty or useful for anything except target
practice in my eye. If anyone has any creative ideas let me know. I was thinking of making a geeky
suit of armor but that's about it.

let me know if you have any ideas.

cool and grey in Ithaca

jg

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We have a Multi-Scan Camera System complete, Model 794 (no computer)
including all wiring, connections and original manual, all in good
order, removed from a CM300 TEM if interested please contact me.

==============================Original Headers==============================
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I bet there is a starving local artist that will snap them up! Try a local art college.

Cheers
Naomi

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Title-Subject: [Filtered] What to do with film plates and cassettes?

Message: When I got my monochromated F20 in 2003 it came with 200 plates and 4 cassettes. I kept
them....just in case. Well, just in case never happened and now I have all this brand-stinking new
and useless scope stuff cluttering up the room. If anyone wants them, god bless you, give me your
fedx number and enjoy. In lieu of that, they're not pretty or useful for anything except target
practice in my eye. If anyone has any creative ideas let me know. I was thinking of making a geeky
suit of armor but that's about it.

let me know if you have any ideas.

cool and grey in Ithaca

jg

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I have to admit, I've always want to see if I could make mobiles from them but snake scales also come to mind.

Stay safe.........

Frank

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Sent: Tuesday, June 04, 2013 10:03 PM
To: Frank Karl

I bet there is a starving local artist that will snap them up! Try a local art college.

Cheers
Naomi

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Title-Subject: [Filtered] What to do with film plates and cassettes?

Message: When I got my monochromated F20 in 2003 it came with 200 plates and 4 cassettes. I kept
them....just in case. Well, just in case never happened and now I have all this brand-stinking new
and useless scope stuff cluttering up the room. If anyone wants them, god bless you, give me your
fedx number and enjoy. In lieu of that, they're not pretty or useful for anything except target
practice in my eye. If anyone has any creative ideas let me know. I was thinking of making a geeky
suit of armor but that's about it.

let me know if you have any ideas.

cool and grey in Ithaca

jg

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Email: ludek,lovicar-at-ist.ac.at
Name: Ludek Lovicar

Organization: IST Austria

Title-Subject: [Filtered] Uranyl acetate

Message: Dear All,
please,could anyone provide me with information from which company in Europe is possible to order
uranyl acetate?

Thank you!
Best regards,
Ludek

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Email: achristensen-at-annamaria.edu
Name: Arne

Organization: Anna Maria College

Title-Subject: [Filtered] AO Spencer dissecting scope alignment

Message: Dear listeners,

I'm trying to align the prisms on an AO Spencer binocular cycloptic dissecting scope. It's been a
frustrating effort because the prisms are glued to the frame, so readjusting them is not trivial.
More frustrating is that the microscope needs to be disassembled, then fully reassembled, before
proper alignment can be determined. Is anyone aware of an old user manual?

Here is a picture of the model:

http://tinyurl.com/mr5d9b9

Thanks,
Arne

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Dear All,

As the deadline for early registration for this year’s Microscopy and
Microanalysis Meeting (Aug. 4-8 2013 in Indianapolis) is coming
closer, I would like to make you aware of our Sunday Short Course on
3D electron microscopy (description below). Please forward this also
to your colleagues who may be interested in this topic.
This course is ideal for people who need a comprehensive introduction
or refresher in 3D reconstruction methods and is suitable for
biologists and material scientists.
If you have already registered for the meeting you can add the course
registration as “extra item only”.

X12 3D Electron Microscopy of Macromolecular Assemblies
Teresa Ruiz, Michael Radermacher, Edward Morris
8:30 AM to 5:00 PM on Sunday, August 4.

This short course will provide a comprehensive description of the
methods used for 3D structure determination from electron micrographs
of macromolecular complexes or weakly scattering specimens available
in multiple copies. Specimen-preparation techniques for single
particles (deep stain, vitreous ice) will be presented, followed by
selection of optimal imaging conditions, including low-dose imaging.
Next, a detailed explanation of image-processing techniques, with
special emphasis on the random-conical reconstruction technique, will
be presented. Finally, structure interpretation and docking of X-ray
structures to 3D EM densities will be demonstrated. The techniques
described could be applied to both biological and materials science
specimens.

--
Michael Radermacher, Ph.D., Prof.
University of Vermont
Dept. Molecular Physiology and Biophysics
HSRF 120 / 149 Beaumont Ave
Burlington, VT 05405
USA

Tel: + 802 656-4834
Fax: + 802 656-0747

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Dear Ludek,

Polysciences sells uranyl acetate
(http://www.polysciences.com/Catalog/Department/Product/98/categoryid--52/productid--1262/search--uranyl_20acetate/)

Best,

Stefan

} Email: ludek,lovicar-at-ist.ac.at
} Name: Ludek Lovicar
}
} Organization: IST Austria
}
} Title-Subject: [Filtered] Uranyl acetate
}
} Message: Dear All,
} please,could anyone provide me with information from which company in Europe is possible to order
} uranyl acetate?
}
} Thank you!
} Best regards,
} Ludek

==============================Original Headers==============================
8, 22 -- From stefan.geimer-at-uni-bayreuth.de Wed Jun 5 08:22:05 2013
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Hi Arne,

The microscope worked at one time, and I seriously doubt that the glued
prisms have shifted. I believe you need to look elsewhere in the
mechanism for the fault. In our similar microscopes, the lubrication
would dry out, and the zoom or ocular space adjustment would not function
properly. You may have already done this, but a thorough cleaning and
re-lubrication usually corrected the problem.

I hope this helps.
Regards,
Darrell

microscopylistserver-noreply-at-microscopy.com wrote on 06/05/2013 08:47:10
AM:

} From: microscopylistserver-noreply-at-microscopy.com
} To: Darrell Miles/Fishkill/IBM-at-IBMUS,
} Date: 06/05/2013 08:48 AM
} Subject: [Microscopy] viaWWW:AO Spencer dissecting scope alignment
}
}
}
}
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} Email: achristensen-at-annamaria.edu
} Name: Arne
}
} Organization: Anna Maria College
}
} Title-Subject: [Filtered] AO Spencer dissecting scope alignment
}
} Message: Dear listeners,
}
} I'm trying to align the prisms on an AO Spencer binocular cycloptic
} dissecting scope. It's been a
} frustrating effort because the prisms are glued to the frame, so
} readjusting them is not trivial.
} More frustrating is that the microscope needs to be disassembled,
} then fully reassembled, before
} proper alignment can be determined. Is anyone aware of an old user
manual?
}
} Here is a picture of the model:
}
} http://tinyurl.com/mr5d9b9
}
} Thanks,
} Arne
}
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Hi Ludek,

We at TAAB Laboratories are able to supply uranyl acetate. Please see our
web site www.taab.co.uk

Best regards

Terry Cooper
TAAB Laboratories Equipment
3 Minerva House, Calleva Park
Aldermaston, Berks, RG7 8NA, England
Tel ++44(0)118 981 7775
Fax ++44(0)118 981 7881
e-mail sales-at-taab.co.uk
www.taab.co.uk

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Greetings Fellow Microscopists,

Two advanced scanning electron microscopy short courses will be instructed by Dr. Craig Schwandt of The McCrone Group at Hooke College of Applied Sciences, located in Westmont, IL this July.

INS-605: ADVANCED X-RAY MICROANALYSIS BY EDS This is an advanced course in x-ray microanalysis using energy-dispersive x-ray spectrometry (EDS). The course will provide instruction and hands-on practice on performing EDS, analyzing difficult samples, mapping, and interpreting analysis results.
http://www.hookecollege.com/courses/course.asp?COURSE_ID=129

INS-610: ADVANCED IMAGING TECHNIQUES FOR SCANNING ELECTRON MICROSCOPY This Scanning Electron Microscopy (SEM) imaging course is an advanced topics course and provides instruction and hands-on practice for getting the best possible SEM images, especially from difficult samples, or under challenging operating conditions. Signals and image generation, instrument operation, operating variables, image interpretation and applications of SEM will be studied through lectures and hands-on activities. Advanced SEM imaging topics such as very high resolution imaging and low voltage imaging will be covered.
http://www.hookecollege.com/courses/course.asp?COURSE_ID=127

Best regards-

__________________________________________________
Chris Gorman
Hooke College of Applied Sciences
850 Pasquinelli Drive
Westmont, IL 60559-5539 USA
630-887-7100 (tel)
630-887-7412(fax)
www.hookecollege.com

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Hi

We are a University lab and can not afford Amira until the next grant Cycle. However we have some nice tilt series acquired on our Titan TEM. We also have some nice reconstruction files from Inspect 3d. However the inspect 3d software does a poor job visualizing the reconstruction files .rec files.

I see the free reconstruction software on the Mathworks website: "CT reconstruction package". Does anybody have experience with this Matlab executable? Does anybody know of other free software? My friend Dudley told me of some free Software written by Mike Stowell of CU Boulder?

Thanks

Pete Eschbach
Oregon State University
541 737 5645

Sent from my iPhone

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Link to images: https://www.dropbox.com/l/OTFitw76NyS2C0DvQQivkc

Lipofuscin, lysosomes, or lipidosis? I guess I should know what these
structures are, but I've never had so many show up and in so many
different tissues until a few of months ago. Are my osmiophilic,
electron-dense blobs a normal part of lipid metabolism? I was looking at
tumors from coral heads that are downhill from a dump in a Pacific island,
and the thing that jumped out was all this dense stuff all over the place,
including in the symbiotic zooxanthellae. Some of these things look a
little like junk next to cholesterol crystals we see in an atherosclerotic
mouse. Soon after I got the coral tumors, I had similar structures crop
up in some cultured HEK cells (that got annoyed when we tried to get them
off the dish live with no trypsin; re-doing that experiment later today),
and I also see them in an undescribed marine eukaryote and in my sick
snails. In my Dropbox you can see:

Three more images of the mystery tubular structure from the snail digestive
gland, showing that they can be very long, and also showing some
lipid-y-looking structures; a cholesterol crystal in mouse artery plaque;
four images from the cultured cells, some showing "fingerprint" structures
associated with the mystery stuff; seven of the coral tumors, including one
showing that their zooxanthellae have it, too; an undescribed marine
eukaryote, and a low mag of the sick snail digestive gland. Most of these
are unstained - the stuff is very electron dense.

So - Lipofuscin, secondary lysosomes, something else? Are these all the
same, or different? And how do fats form crystals? Are crystals likely to
be cholesterol (animal studies), or can other fats crystalize (food
science, like ice cream)?

Here's the link, again:
https://www.dropbox.com/l/OTFitw76NyS2C0DvQQivkc

Aloha,
Tina

--
Tina Weatherby Carvalho
University of Hawaii
Pacific Biosciences Research Center
Biological Electron Microscope Facility
2538 McCarthy Mall
Honolulu, HI 96822
808-956-6251

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Email: acalhoun-at-bidmc.harvard.edu
Name: Andrea Calhoun

Organization: Beth Israel Deaconess Medical Center

Title-Subject: [Filtered] How To: Cryogens for Ambient Pressure Plunge/Slam Freezing

Message: Ok Listservers; I've had enough fishing in the dark. All I need is an answer. I'm going to
swallow my pride and hope that you'll understand my cautious safety inquiry.

How do you get ethane into a small copper cup for plunge freezing!?!

This may seem silly, but I have no experience with ethane or propane in this regard. I Get that the
liquid ethane is under pressure and comes out as a gas. I expect the gas to eventually liquefy if
the surrounding environment is cooled with LN2. This is correct, right?

My Environmental Health and Safety guy thought I was nuts and recommended I use LN2 or cooled
isopentane (though all the publications I SENT him say otherwise) and the people from Airgas do "not
recommend using ethane or propane for this type of application due to the flammability of these
gases". Also, Google search fail.

I basically need to know:

1) if there is a more technical term when asking for a 'lecture' size cylinder

2) if there is a special nozzle, regulator, or tubing that I will need

3)if I am correct about the whole 'pouring' ethane gas into a LN2 cooled copper cup

Please send all videos, publications, how-to methods, typed information, website URLs-- I'm
desperate here! And I know its so simple! AND, most importantly, I want to be safe!

Andrea Calhoun
Electron Microscopy Specialist
P: 617-667-4188
E: acalhoun-at-bidmc.harvard.edu

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Email: r.gilbert-at-auckland.ac.nz
Name: Ray Gilbert

Organization: University of Auckland

Title-Subject: [Filtered] Leica DM IRB board

Message: Hi

One of our Leica DM IRB microscopes has an issue with its board and it needs replacing. Our
Facilities Administrator can't source a replacement.

Does anyone have any idea where I could find a replacement for this?

Thanks

Ray Gilbert
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Centre for Brain Research
University of Auckland

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Email: akolmakov-at-physics.siu.edu
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Title-Subject: [Filtered] exchange Hitachi S4500 FE gun

Message: Dar colleagues,
we have been donated used Hitachi S4500 FEG SEM more than two years ago and now FEG needs to be
changed. Unfortunately we do not have exchange kit which is usually supplied with the microscope.
The list is below.
In case somebody has it, we will be very thankful to be able to borrow it (if possible) for
two-three weeks in coming August? The single use spare parts we can purchase. Postal expenses will
be covered. Thank you very much in advance
Andrei Kolmakov
Associate Professor
Department of Physics,
Southern Illinois University
1245 Lincoln Dr., Neckers 491
SIUC, Carbondale IL 62901 USA
Phone office (618)-453-5212
Phone 407 lab (618)-453-6106
Phone 140 lab (618)-453-6011
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The standard kit includes:
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Quantity needed 1
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15) Spacer
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Some part numbers may have been superseded with new numbers.

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Email: zhangyc-at-gmail.com
Name: Yucheng Zhang

Title-Subject: [Filtered] Carbon contamination of semiconductor samples in STEM

Message: Hi,
My semiconductor TEM samples (mainly Ge and GaN etc.) seem to always suffer from carbon
contamination in STEM analysis. The cross-section samples were prepared using a conventional way,
i.e. mechanical polishing and Ar-ion milling. To minimize the contamination, I have tried different
glues including Araldite epoxy and M-bond 610 adhesive to glue the sandwich cross-sections. Before
the STEM analysis, the sample was always cleaned with an Ar/O plasma-cleaner. Soaking the sample in
acetone and baking were also tried, but none of the procedures seems to avoid carbon depositing
quickly on the surface in STEM. No chemical analysis can be performed due to the carbon deposition.
Just wondering if anybody has had a similar experience and a solution to the problem.
Many thanks.

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Email: Z.Zhou-at-lboro.ac.uk
Name: Zhou

Organization: Loughborough University

Title-Subject: [Filtered] TEM - quantification phase by diffraction Intensity

Message: Ive got a two-phase composite sample (one is amorphous). The TEM electron diffraction
patterns show halos (from the amorphous) plus the usual spots (from the crystal phase). Patterns are
recorded by CCD camera.

The questions are:
1, how reliable is it to quantify the amount of amorphous materials by comparing the intensity of
the halos in the electron diffraction patterns? Is there a direct relation between the concentration
of a phase in a composite with the intensity of its diffraction pattern?
2, how does sample thickness affect the patterns?
3, are there any issues to do with the sensitivity limit of CCD camera when the amorphous halos are
significantly weaker than the crystal spots?

Zhaoxia Zhou

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Zhou,

I'd rather you get the mass fractions from spectroscopy rather than
diffraction, as you say thickness can have complicated effects in
anything but a two-beam condition for the crystalline phase.

The other day I had some 1-2nm silver nano-particles that gave two fuzzy
rings only, and yet using dark field imaging I could make out individual
crystallites, and in bright field there was obvious diffraction contrast.

Having said that I'd want to say there is a relationship between
concentration and intensity, but the crystals might produce diffuse
scattering too (increasingly with thickness) which could be confusing. A
colleague (Graeme Raynerd) once measured electron scattering from cold
gases in a TEM as part of an advanced Physics degree - lowering the
temperature of your materials could help reduce the Debye-Waller factor.

Good Luck!

Rob Keyse
Lehigh

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} Name: Zhou
}
} Organization: Loughborough University
}
} Title-Subject: [Filtered] TEM - quantification phase by diffraction Intensity
}
} Message: Ive got a two-phase composite sample (one is amorphous). The TEM electron diffraction
} patterns show halos (from the amorphous) plus the usual spots (from the crystal phase). Patterns are
} recorded by CCD camera.
}
} The questions are:
} 1, how reliable is it to quantify the amount of amorphous materials by comparing the intensity of
} the halos in the electron diffraction patterns? Is there a direct relation between the concentration
} of a phase in a composite with the intensity of its diffraction pattern?
} 2, how does sample thickness affect the patterns?
} 3, are there any issues to do with the sensitivity limit of CCD camera when the amorphous halos are
} significantly weaker than the crystal spots?
}
} Zhaoxia Zhou
}
}
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--
Robert Keyse
EM Facility
Whitaker Laboratory
5 East Packer Avenue
Bethlehem
PA 18015
USA

Tel. +1 610 758 3465
Fax +1 610 758 4244

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Hello All,
Recently I have been attempting to embed some E.coli with LR White
with intentions to immunogold label some thin sections however the
protocol I am using is not producing well embedded bacteria. When I
view the sectioned bacteria with a TEM there are significant gaps
between the bacteria and the plain resin.
I have already increased the time the sample is rotating in 100% LR
white which has marginally improved the embedding. The protocol is
listed below. Is it possible the bacteria need to be fixed for a
longer period of time? If a longer fixation is needed, at what point
might the fixation be a detriment to the eventual labeling? Thanks in
advance for any advice and help

-Joe

Protocol:
-Fix bacteria with 1% glutaraldehyde + 1% paraformaldehyde in 0.1M
PIPES, 2 Hr at 4oC
-Wash in buffer overnight at 4oC
-50% Ethanol, 15 min(sample rotating from here on out)
-70% Ethanol, 15 min
-80% Ethanol, 10 min
-2:1 LR White: 80% Ethanol,1 hour
-100% LR White, 1hour
-100% LR White, overnight
-100% LR White, overnight
-100% LR White, 30 min
-Place sample is gelatin capsules and heat at 50oC for 24hr.

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Hi, Joe-

I typically follow the 80% ethanol with 95%, two changes, and then three
changes of 100% (fresh, absolute) ethanol of 10 minutes each. Then 1:1
absolute ethanol LR White overnight, then into the 100% LR White changes.
I know LR White is supposed to be able to handle some water, but I don't
believe it. We're pretty anal about water/humidity here in beautiful Manoa
Valley.

Aloha,
Tina

} Recently I have been attempting to embed some E.coli with LR White
} with intentions to immunogold label some thin sections however the
} protocol I am using is not producing well embedded bacteria. When I
} view the sectioned bacteria with a TEM there are significant gaps
} between the bacteria and the plain resin.
} I have already increased the time the sample is rotating in 100% LR
} white which has marginally improved the embedding. The protocol is
} listed below. Is it possible the bacteria need to be fixed for a
} longer period of time? If a longer fixation is needed, at what point
} might the fixation be a detriment to the eventual labeling? Thanks in
} advance for any advice and help
}
} -Joe
}
} Protocol:
} -Fix bacteria with 1% glutaraldehyde + 1% paraformaldehyde in 0.1M
} PIPES, 2 Hr at 4oC
} -Wash in buffer overnight at 4oC
} -50% Ethanol, 15 min(sample rotating from here on out)
} -70% Ethanol, 15 min
} -80% Ethanol, 10 min
} -2:1 LR White: 80% Ethanol,1 hour
} -100% LR White, 1hour
} -100% LR White, overnight
} -100% LR White, overnight
} -100% LR White, 30 min
} -Place sample is gelatin capsules and heat at 50oC for 24hr.
}

****************************************************************************
* 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*
****************************************************************************

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Email: duraine-at-bcm.edu
Name: Lita Duraine

Organization: Howard Hughes Medical Institute

Title-Subject: [Filtered] RE: Issue with Embedding LR White

Message: Hi Joe,

I agree with Tina. We also continue the graded series of EtOH up to 100%, followed by 1:1 ratio and
then 3 x in pure LR White. We do Immuno routinely with LR White and get good results.

I have found that the LR White needs to be somewhat fresh (within 6 months) to work better. We also
use vacuum steps for our fixation and infiltration which helps significantly.

Lita

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Hi Joe,

I agree with Tina. We also continue the graded series of EtOH up to 100%, followed by 1:1 ratio and then 3 x in pure LR White. We do Immuno routinely with LR White and get good results.

I have found that the LR White needs to be somewhat fresh (within 6 months) to work better. We also use vacuum steps for our fixation and infiltration which helps significantly.

Hope this helps,

Lita Duraine
Electron Microscopist
Bellen Lab
HHMI- Molecular Genetics
Duncan Neurological Research Institute
1250 Moursund St.
Houston, TX 77030
Rm: N1165.17
MS: N1125.50
832-824-8772 TEM Room
979-549-6526 Cell
http://flypush.imgen.bcm.tmc.edu/lab/people/lita.php

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If you are after something fancy like Amira then the "CT reconstruction package"
may not be for you. You might find ParaView more "amenable" ( http://www.paraview.org/ )
However, for a ready to go solution I'd recommend VolView (http://www.kitware.com/opensource/vvdownload.html )

You may want to explore the combination (ImageJ + TomoJ) + (Fiji + TrakEM2).
TomoJ is plugin developed specifically oriented towards TEM electron tomography
and TrakEM2 allows morphological data mining, three-dimensional modelling and
image stitching, registration, editing and annotation of EM sets. You will find
tons of literature and howtos on the net.

Good luck!

-----Original Message-----
X-from: peter.eschbach-at-comcast.net [mailto:peter.eschbach-at-comcast.net]
Sent: Wednesday,05 June 2013 22:07
To: CHINEA CANO, Ernesto

Hi

We are a University lab and can not afford Amira until the next grant Cycle. However we have some nice tilt series acquired on our Titan TEM. We also have some nice reconstruction files from Inspect 3d. However the inspect 3d software does a poor job visualizing the reconstruction files .rec files.

I see the free reconstruction software on the Mathworks website: "CT reconstruction package". Does anybody have experience with this Matlab executable? Does anybody know of other free software? My friend Dudley told me of some free Software written by Mike Stowell of CU Boulder?

Thanks

Pete Eschbach
Oregon State University
541 737 5645

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Message: I just wanted to say Thank You! to all the helpful detailed
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-------- Original Message --------

Dear Collective,

Just a fishing expedition here: Is anybody out there having unusual problems with poor infiltration of resin into a variety of tissues in recent weeks? If so, can we compare notes?

Cheers,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211

Phone: 573-884-5383
Email: tindallr-at-missouri.edu
Day job: www.emc.missouri.edu
Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
Blog: http://nadiasyard.com

==============================Original Headers==============================
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9, 28 -- Subject: TEM: Infiltration problems, anybody?
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Hi all,

We have some students rehabbing a Porter-Blum MT2-B microtome (also a
MT1) and an LKB knifemaker. Does anyone have the service manuals? How
about the owners books?

Other than cleaning the old grease and replacing the O-ring drive belts,
are there common problems to look out for?

Thanks!
Henk

--
Untitled Document

The Ohio State University
Hendrik O. Colijn
*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*
1305 Kinnear Rd, Suite 100, Columbus, OH 43212

614.643.3458 Office colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu}
www.cemas.osu.edu {http://www.cemas.osu.edu}

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

==============================Original Headers==============================
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Hi!

Does anyone have some Cupromeronic Blue left? Or knows where I can buy
some? Unfortunately all the companies we contacted say they do not have
it any more. We would need about 100 mg, or better 500mg.

Thanks!
Stefan

==============================Original Headers==============================
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Henk,

I have both, and if I remember rightly, as pdfs.
But .... I'm at the LeHigh school this week, and can't send them until next Monday. If you don't get any before then, please let me know and I'll send what I have.

Phil

Philip Oshel
Supervisor, Biology Dept. Microscopy Facility
Central Michigan University
________________________________________
X-from: colijn.1-at-osu.edu [colijn.1-at-osu.edu]
Sent: Monday, June 10, 2013 9:13 PM
To: Oshel, Philip Eugene

Hi all,

We have some students rehabbing a Porter-Blum MT2-B microtome (also a
MT1) and an LKB knifemaker. Does anyone have the service manuals? How
about the owners books?

Other than cleaning the old grease and replacing the O-ring drive belts,
are there common problems to look out for?

Thanks!
Henk

--
Untitled Document

The Ohio State University
Hendrik O. Colijn
*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*
1305 Kinnear Rd, Suite 100, Columbus, OH 43212

614.643.3458 Office colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu}
www.cemas.osu.edu {http://www.cemas.osu.edu}

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

==============================Original Headers==============================
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==============================Original Headers==============================
19, 35 -- From oshel1pe-at-cmich.edu Tue Jun 11 10:52:09 2013
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19, 35 -- From: "Oshel, Philip Eugene" {oshel1pe-at-cmich.edu}
19, 35 -- To: "colijn.1-at-osu.edu" {colijn.1-at-osu.edu}
19, 35 -- CC: "microscopy-at-microscopy.com" {microscopy-at-microscopy.com}
19, 35 -- Subject: RE: [Microscopy] Porter-Blum MT2-B & LKB Knifemaker manuals
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We tried to get some Cupromeronic Blue from the following companies:

SIGMA-Aldrich, Polyscience, Santa Cruz Technologies, USBiological,
Sanova and also Stratech.

but we will try it at creschem and thrivechem, thanks for the help, John!

We normally try to avoid toxic substance in our lab when ever it is
possible, but unfortunately there is no alternative substance known for
Cupromeronic Blue to carry out our experiment.

Stefan

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Announcing the 4th Annual Scientific Imaging and Quantization course in
Minneapolis, MN, August 26 - 29, 2013.

I am giving this 3 - 4-day course as a way to meet a personal mission,
which includes:

1. Providing instruction on proper methods for working with images and
image stacks.
2. Providing means for making computer-aided quantitation simpler and
more consistent with complex biological images.
3. Showing pro-active means for maintaining image integrity throughout
the microscope slide to publication process.

To learn more about the course, please go to the website
quickphotoshop.com, or to
http://imagingandanalysis.com/4th_annual_seminars-withmovies.html
You can do that right now. Really.

Or read on...

The class is for anyone working with scientific imaging using
microscopes. The only requirement is that you are able to find files on
your hard drive with relative ease, and that you have fairly good
hand-eye coordination with the use of a mouse (or your dexterous fingers).

You will receive a certificate of completion that can be included in
your CV. You can bring along images from which you would like to get
measurements.

You will learn about the following using methods with techniques you
have not likely learned (e.g., how to see if you have saturated pixels
when using the Levels tool):

Image Integrity: what you can, can't and should do.
Calibration of Monitors (NEW!)
Calibration devices and techniques (NEW!)
Digital Imaging Terms (e.g., gamma, gamut, etc)
Opening 12-bit images (10-bit, 14-bit too) in 16-bit space
Opening movies and image stacks
Photostitching (montaging several images together)
Z-projection in Photoshop
Cropping, Rotating, Flipping
De-Noising methods (averaging single pixel outliers, median filtering,
surface blur, etc)
Uneven Illumination Correction (flatfield correction, using image
itself, vignetting, etc.)
Tonal/Color Correction (manual, linear histogram matching, histogram
matching, etc.)
Co-existence ("colocalization") method
Merging
Colorizing/DeColorizing
Working with movies and lettering/adding symbols
Creating Automated Macros (Actions) to apply to a folder of images
Creating Figures/Plates: In Illustrator and Photoshop
Adding Lettering/Symbols
Making Inserts
To Outputs: Publication, Laptop Projector, Grants, Posters, Powerpoint
How to Use the Image Size function for outputting
Sharpening (2 methods)
Quantization: Segmenting (separating out objects of interest from
surrounding areas)
Segmenting color images
Typical Method for Measuring Counts
Typical Method for Measuring Morphometry (surface area, lengths,
clustering, etc)
Methods for Measuring Intensity/Density
Intro to Stereology
Assistance with your Images
Hands on Lab day (day 4)

Here's what you will need to bring along:

1) Bring a laptop with (preferably) Photoshop versions CS3 - CS6:
earlier versions (Photoshop 6 - CS2) will be OK, but a limited number of
functions will not work in Photoshop 6 - CS2.
2. You can bring a Mac or PC.
3. Bring a mouse, unless you are super-dexterous with your fingers.

Please write to me with any questions: jerry-at-imagingandanalysis.com.

All the best!

Jerry Sedgewick

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Email: gregory.pooley-at-y7mail.com
Name: Dr Gregory D Pooley

Organization: CMCA University of Western Australia

Title-Subject: [Filtered] Zeiss SEM

Message: I'm interested in locating and purchasing a secondhand R Series Zeiss ParticleScan,
sometimes referred to as a QEMSCAN. Post 1995 or more recent will do. Hopefully the instrument is
in reasonable condition or at least salvageable.

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I wanted to give a public thank you to the generous members of the
listserver who shared their experiences & documentation with us. It is a
pleasure to be associated with such generous and collegial people!

Among the respondents were:
Wai Pang Chen
Wolfgang Muss
Jason Saredy
James Ehrman
Ed Haller
Pat Connelly
Timothy Morken
Phil Oshel
Mike Pendleton
Casey Peto

Cheers,
Henk

At 6/10/2013 9:10 PM, colijn.1-at-osu.edu wrote:
}
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} Hi all,
}
} We have some students rehabbing a Porter-Blum MT2-B microtome (also a
} MT1) and an LKB knifemaker. Does anyone have the service manuals? How
} about the owners books?
}
} Other than cleaning the old grease and replacing the O-ring drive belts,
} are there common problems to look out for?
}
} Thanks!
} Henk
}
}

--
Untitled Document

The Ohio State University
Hendrik O. Colijn
*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*
1305 Kinnear Rd, Suite 100, Columbus, OH 43212

614.643.3458 Office colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu}
www.cemas.osu.edu {http://www.cemas.osu.edu}

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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Email: jpshield-at-uga.edu
Name: John Shields

Organization: University of Georgia

Title-Subject: [Filtered] Position for Lab Manager at EM Lab

Message: Title: LABORATORY MANAGER II
Posting Number: 20130724
Department: 275-Electron Microscope Laboratory

The University of Georgia's Center for Advanced Ultrastructural Research (CAUR) is seeking a
scientist in materials and physical characterization at the M.S. level.

This individual will collaborate with physical sciences users in experimental design, implementation
and analysis.
The individual should have experience with SEM, TEM, electron diffraction and associated sample
preparation techniques. They will also be responsible for training and supervising users in TEM,
SEM, electron diffraction, and x-ray spectroscopy analysis. Experience in instruction for course
offerings such as Electron Microscopy, Techniques in Modern Microscopy, and Clay Mineralogy is optional.

The individual will assist materials/physical sciences users of the CAUR and share responsibility
for the maintenance of instruments including two TEMs and two SEMs and all sample preparation
equipment.

Entry level salary will be approximately $47,000.00, for a 12 month appointment, with an increase
commensurate with demonstrated experience.

Inquiries and applications should be directed to Dr. Paul A. Schroeder, Center for Advanced
Ultrastructural Research, 151 Barrow Hall, University of Georgia, Athens, GA 30602 USA.
(706)542-4080. caur-at-uga.edu.

Please include curriculum vitae, statement of interest, and contact information for at least three
references.
The University of Georgia is an Equal Employment Opportunity/Affirmative Action Institution.

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Email: jacqueline.ayotte-at-ticona.com
Name: Jackie Ayotte

Organization: Ticona Engineering Polymers

Title-Subject: [Filtered] Precise 3D Measurement Capability Needed

Message: Dear Listers,

Does anyone use equipment, or has anyone observed equipment that can measure X, Y, Z directions of
small (~8x8x2mm) parts with 1um accuracy (or less 0.5um) with very low tolerance?

If so, please respond at your (very) earliest convenience.
If possible please include equipment name, manufacturer, and model number.

With Many Thanks,

Jackie Ayotte
Advanced Materials Characterization
Senior Microscopist

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Email: h.turner-at-waikato.ac.nz
Name: Helen Turner

Organization: University of Waikato

Title-Subject: [Filtered] Condenser apertures

Message: Hello All,

We have a relatively old Philips CM30 here at Waikato. We are having
problems with dirt on the condenser apertures. They are the old 'solid'
type, 3mmx3mm and they are going to be flamed shortly.
I was wondering, should this fail, whether anyone out there should happen
to have any apertures of this sort? The sizes are 30, 50,100 and 200um.

Many thanks,

Helen.

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Organization: Monash Centre for Electron Microscopy

Title-Subject: [Filtered] Two Electron Microscopist positions vacant at the Monash Centre for
Electron Microscopy

Message: Applications are invited for two Electron Microscopist positions at the Monash Centre for
Electron Microscopy, Monash University, Melbourne, Australia (www.mcem.monash.edu.au), as follows:

Electron Microscopist LEVEL HEW 8 - Continuing Appointment
Job No. 512238
Employment Type: Full Time

Remuneration: AU$97,129 - $107,211 pa HEW Level 08
(includes 17% employer superannuation)

Electron Microscopist LEVEL HEW 7 - Continuing Appointment
Job No. 513488
Employment Type: Full Time.
Remuneration: AU$86,334 - $94,693 pa HEW Level 07
(includes 17% employer superannuation)

Details at: http://jobs.monash.edu.au/ and search relevant Job No. or Electron Microscopist

Contact: Professor Joanne Etheridge, Director, Monash Centre for Electron Microscopy
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Email: stefan.geimer-at-uni-bayreuth.de
Name: Stefan Geimer

Organization: Universit�t Bayreuth

Title-Subject: [Filtered] LKB Sj�strand Ultra-Microtome Type 3314A

Message: Dear all,

I recently got hands on a LKB Sj�strand Ultra-Microtome Type 3314A (it's from the fifties)and really
would like to get it up and running. The microtome seems to be pretty complete, a binocular (in case
it came with one) is missing, but it should be no problem to improvise something. However, I can't
find any informations about that really beautiful machine on the internet, so I am desperately
looking for a manual or whatever material dealing with the LKB 3314A. Is there anybody out there who
has a manual or whatever material(catalogues, ads)and would be willing to make me a paper copy or
scan. Of course I would reimburse any accuring costs for copies and postage.

Best,

Stefan

*****************************************
Stefan Geimer, Ph.D.
Universitaet Bayreuth
Biologie/Elektronenmikroskopie NW I / B1
Universitaetsstr. 30
95447 Bayreuth
Germany

stefan.geimer-at-uni-bayreuth.de
*****************************************

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Email: thomas.beck-at-novelis.com
Name: TJ Beck

Organization: Novelis

Title-Subject: [Filtered] Electron microscopy short courses

Message: Does anyone know of good short courses in electron microscopy in the Atlanta area? I am
specifically looking at improving my knowledge and understanding in the area of EDS, EBSD, and FIB.

Thanks,

TJ

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Dear Stefan,
unfortunately the only web sources I could find for now were:

Perhaps there possibly are available archive copies at LEICA (former REICHERT),
which had incorporated LKB in the late 80ies?

http://www.lkbprod.com/test/pdf/UM_Timeline.pdf: German info....LKB 3414A: 1955&
http://www.lkbprod.com/test/pdf/UM_Timeline.pdf : English, info loaded via:
http://www.sputtr.com/read/the-lkb-ultramicrotomy-products-d41d.html?f=1qeXpurpn6Wih-SUpOGumaanh8DX24XAs6eQx9Xi5sfi1sje3urU4eGF4OTY0unJ6eCHp-Kwlq6Ky5Kt3Kiona-PzeDj5p-jl9zn6Zfa38jl39TQndnU4ZfZ1eXdneTK25y6uc7KzuHN0dngzpzkytuPoOk

LKB 2218 Historange:
http://www.google.at/url?sa=t&rct=j&q=lkb%20and%203314a%20and%20microtome&source=web&cd=2&cad=rja&ved=0CDQQtwIwAQ&url=http%3A%2F%2Fwww.youtube.com%2Fwatch%3Fv%3D6ZBHveyKEnI&ei=xC-8Ue3vGImqO9LrgKgP&usg=AFQjCNETuVGaJuyq-k1I61zjdQR8PXbSjw

LKB 2128 Ultrotome IV Ultramicrotome:
http://websites.labx.com/rankin/detail.cfm?autonumber=9290

Best wishes and good luck,

Wolfgang

Von: microscopylistserver-noreply-at-microscopy.com [mailto:microscopylistserver-noreply-at-microscopy.com]
Gesendet: Samstag, 15. Juni 2013 02:00
An: Mu� Wolfgang
Betreff: [Microscopy] LKB Sj�strand Ultra-Microtome Type [Manual??? - LKB Sj�strand Ultra-Microtome Type 3314A from the 1950ies]

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Dear all,
I recently got hands on a LKB Sj�strand Ultra-Microtome Type 3314A (it's from the fifties)and really
would like to get it up and running. The microtome seems to be pretty complete, a binocular (in case
it came with one) is missing, but it should be no problem to improvise something.
However, I can't find any informations about that really beautiful machine on the internet, so I am desperately
looking for a manual or whatever material dealing with the LKB 3314A.
Is there anybody out there who has a manual or whatever material (catalogues,ads)and would be willing to make me a paper copy or scan. Of course I would reimburse any accuring costs for copies and postage.

Best,

Stefan

*****************************************
Stefan Geimer, Ph.D.
Universitaet Bayreuth
Biologie/Elektronenmikroskopie NW I / B1
Universitaetsstr. 30
95447 Bayreuth
Germany

stefan.geimer-at-uni-bayreuth.de
*****************************************

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Email: elizabeth.barrios-at-nasa.gov
Name: Elizabeth

Organization: NASA (Intern)

Title-Subject: [Filtered] User Documents for Gold Coater

Message: Hello! I am currently an intern working on thermal transport through polymers this summer
at NASA Glenn Research Center. We want to utilize a gold coater that was purchased and used in the
early 2000's but we cannot locate any sort of operation manuals.

THe product was an Ernest F. Fullam Gold Coater with serial number 18930-0805-74.

Does anyone happen to have these documents that they can scan and email or mail to me? (I'd be
willing to pay a nominal fee to retrieve these documents.)

Elizabeth

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Dear All,
I am trying to image some falcon feathers in the SEM. A first look with the Stereo microscope showed a lot of dirt and maybe
grease also on the feathers.
Can somebody please advice me how to clean the feathers and get an as clean as possible and as stable as possible specimen for SEM?
Does it make any sense to use Osmiumtetroxide with feathers to lower the possibility of charging under the e-beam?

Best wishes,
Stefan

--

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www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
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Dear All,
no, I have no ESEM and no low coltage possibility. Concerning the detector signals I need I will have to work with at least 15 kV
and 200 nanometer spotsize, and this for a long time ;-)
If anyone has some experience with specimen preparation in that direction, that would be fine.

Best,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
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Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 17.06.13 21:49, schrieb John Shields:
} I'm not too good at the cleaning part, but unless you have a variable pressure scope those feathers are going to be tough to get
} rid of the charge - that or a very low voltage (ie 1-2 keV)
} John Shields
} UGA Athens GA
}
}
} On Mon, Jun 17, 2013 at 3:26 PM, {stefan.diller-at-t-online.de {mailto:stefan.diller-at-t-online.de} } wrote:
}
}
}
}
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}
} Dear All,
} I am trying to image some falcon feathers in the SEM. A first look with the Stereo microscope showed a lot of dirt and maybe
} grease also on the feathers.
} Can somebody please advice me how to clean the feathers and get an as clean as possible and as stable as possible specimen for
} SEM?
} Does it make any sense to use Osmiumtetroxide with feathers to lower the possibility of charging under the e-beam?
}
} Best wishes,
} Stefan
}
}
} --
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 {tel:%2B%2B49-931-7848700} Phone
} ++49-931-7848701 {tel:%2B%2B49-931-7848701} Fax
} ++49-175-7177051 {tel:%2B%2B49-175-7177051} Mobile
}
} Websites:
} www.nanoflight.info {http://www.nanoflight.info}
} www.stefan-diller.com {http://www.stefan-diller.com}
} www.electronmicroscopy.info {http://www.electronmicroscopy.info}
} www.elektronenmikroskopie.info {http://www.elektronenmikroskopie.info}
} www.zwillingsprojekt.de {http://www.zwillingsprojekt.de}
} www.assisi.de {http://www.assisi.de}
} Anfahrt: http://Mail.map24.com/Stefan.Diller
} -----------------------------------------------------
}
} ==============================Original Headers==============================
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8, 22 -- From stefan.diller-at-t-online.de Mon Jun 17 14:55:57 2013
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Organization: Annashae Staffing

Title-Subject: [Filtered] Flexible Job Opportunity in Electron Microscopy - Cleveland, OH

Message:
Attention health technicians certified to perform Electron Microscopy services:

Annashae Staffing Corporation has the need to fill a part time position for our client, a medical
center in Cleveland, Ohio.

The position will require 1 year of service from start date (may choose to start this year or choose
one of next three years) and will consist of 1,248 hours/year between Monday-Friday 8:00am-4:30pm
with occasional emergency off schedule hours. Schedule is flexible and hours to be worked will be
negotiated upon employment.

Pay rate negotiable at 20-25/hour

Scope of work includes but not limited to:
-Performs fixation, orients very small tissue samples and embeds them in epoxy
-Sections epoxy-embedded patient samples at thickness suitable for both light and electron
microscopic examination
-Stains ultra-thin sections of tissue specimens on metal grids to capture details of the
ultrastructure of cells
-Performs maintenance on equipment including electron microscope, ultra microtome, digital cameras
and advanced computer equipment
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Title-Subject: [Filtered] Zeiss EM9 TEM needs filament change

Message:
We're looking for a local repair person who can provide and change the filament in an old Zeiss EM9
TEM. Can anyone suggest a contact? Please reply off-line to de.wood-at-ars.usda.gov

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Title-Subject: [Filtered] Feathers

Message: I've looked at feathers with the SEM. No problems with dirt or grease. I just sputter
coated them to reduce the possibility of charging. You can also use a lower KV. Try a little
distilled water to rinse them at first to see what that will do to clean them. You could also use a
dilute solution of ETOH/water.

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Hi - I just wanted to announce that Merritt Microscopy is having a
hands-on demo of the Hitachi TM3000 Tabletop Scanning Electron
Microscope, Mondays through Thursdays starting this week, June 17th to
June 20 and next week, June 24 to June 27. The event is open to the
general public, with absolutely no prior microscope experience needed,
and whatever samples you wish to bring. Simply email me to reserve a
time - staff are available by appointment from 2 to 10 pm, and earlier
in the day by special arrangement.

Here's a poster with more information, and a couple of maps giving the
location of the campus. The Demo is on the second floor of the D
building, in Room D243.

http://www.scribd.com/doc/148359131/Hitachi-SEM-Demo
http://goo.gl/maps/v8v4m
http://www.merritt.edu/sites/default/files/campusmap2008.pdf

(Note that the second map is oriented with South on the up side, which
is the reverse orientation of other maps.)

Peter

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Hi Stephan,

Sounds like a feather taken from a drawer, long after it was donated by the bird. When I look at feathers picked up from the grass in my yard, they appear to be clean as whistle.

If you haven't seen these bio-objects before, be informed that only the fact that birds are never awarded patents kept them from being awarded one for the invention of 'vel-crow?'

My recommendation is to clean the feather by dipping it for brief periods only in a dilute solution of really good detergent in a sonicator. Then get some oil akin to that from a bird's preen gland and re-preen it.

Since I have only ever looked at 'fresh' feather in our ESEM at non-charging pressure with HOH gas, I can't give advice for low pressure viewing except to dry after cleaning, and coat lightly with a good conductor. Of course, you are probably welcome to stop by the FEI booth at M&M or stop by our lab and we'll have a look together. No sarcasm intended, please. I have never forgotten my first look at feather with a dissecting scope way back in the 1960's. Hooks and loops!!!! Wow!!!

Cheers,

Hope you are successful with the cleaning.

Fred Monson

Frederick C Monson, PhD
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pannsylvania
West Chester, PA, 1938
610-738-0437
fmonson-at-wcupa.edu
________________________________________
X-from: stefan.diller-at-t-online.de [stefan.diller-at-t-online.de]
Sent: Monday, June 17, 2013 3:23 PM
To: Monson, Frederick

Dear All,
I am trying to image some falcon feathers in the SEM. A first look with the Stereo microscope showed a lot of dirt and maybe
grease also on the feathers.
Can somebody please advice me how to clean the feathers and get an as clean as possible and as stable as possible specimen for SEM?
Does it make any sense to use Osmiumtetroxide with feathers to lower the possibility of charging under the e-beam?

Best wishes,
Stefan

--

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Stefan Diller - Scientific Photography
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D - 97072 Wuerzburg Germany
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Anfahrt: http://Mail.map24.com/Stefan.Diller
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I looked at feathers years ago just for fun. I first, as I remember it, washed the sample in mild soap and water and rinsed well. I reshaped it while wet and gave it a dip in isopropanol (it's what we used in the lab). I let it dry over night. I isolated the portion I wanted to see and gold coated both sides. I mounted with silver paint and ruined several samples until I found the right degree of liquid to dry silver paint. I then gold coated again to insure conductivity.

I wasn't totally happy with the images, but I sure had fun.

I should mention I was playing with flight feathers. Down and body feathers maybe different.

Let us know how it works out.

Frank

-----Original Message-----
X-from: stefan.diller-at-t-online.de [mailto:stefan.diller-at-t-online.de]
Sent: Monday, June 17, 2013 4:05 PM
To: Frank Karl

Dear All,
no, I have no ESEM and no low coltage possibility. Concerning the detector signals I need I will have to work with at least 15 kV
and 200 nanometer spotsize, and this for a long time ;-)
If anyone has some experience with specimen preparation in that direction, that would be fine.

Best,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 17.06.13 21:49, schrieb John Shields:
} I'm not too good at the cleaning part, but unless you have a variable pressure scope those feathers are going to be tough to get
} rid of the charge - that or a very low voltage (ie 1-2 keV)
} John Shields
} UGA Athens GA
}
}
} On Mon, Jun 17, 2013 at 3:26 PM, {stefan.diller-at-t-online.de {mailto:stefan.diller-at-t-online.de} } wrote:
}
}
}
}
} ----------------------------------------------------------------------------
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} Dear All,
} I am trying to image some falcon feathers in the SEM. A first look with the Stereo microscope showed a lot of dirt and maybe
} grease also on the feathers.
} Can somebody please advice me how to clean the feathers and get an as clean as possible and as stable as possible specimen for
} SEM?
} Does it make any sense to use Osmiumtetroxide with feathers to lower the possibility of charging under the e-beam?
}
} Best wishes,
} Stefan
}
}
} --
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 {tel:%2B%2B49-931-7848700} Phone
} ++49-931-7848701 {tel:%2B%2B49-931-7848701} Fax
} ++49-175-7177051 {tel:%2B%2B49-175-7177051} Mobile
}
} Websites:
} www.nanoflight.info {http://www.nanoflight.info}
} www.stefan-diller.com {http://www.stefan-diller.com}
} www.electronmicroscopy.info {http://www.electronmicroscopy.info}
} www.elektronenmikroskopie.info {http://www.elektronenmikroskopie.info}
} www.zwillingsprojekt.de {http://www.zwillingsprojekt.de}
} www.assisi.de {http://www.assisi.de}
} Anfahrt: http://Mail.map24.com/Stefan.Diller
} -----------------------------------------------------
}
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Hi,

Recently our LEO 1450VP SEM behaved oddly and we suspect it is related to
the HV tank.� What happens is that overtime the beam intensity is
diminishing and the image is getting darker and darker.� To restore the beam
intensity, we will have to increase the high voltage.� The column has been
cleaned twice to make sure it is clean.� Another odd thing is that the beam
current reading value is always at maximum (1200 micro amps), not matter the
filament is on or off, or the HV is on or off. We know for sure something is
going wrong, possibly the HT tank.� Would anyone out there have idea what is
happening and how to fix it? Thanks in advance.

Best Regards
---------------------------------
Sean Yang
Regional Analytical Centre
Saint Mary's University
Halifax, NS Canada B3H 3C3
Phone: 902-420-5709
http://www.smu.ca/institutes/emc/welcome.html
http://www.smu.ca/institutes/rgc/welcome.html

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Email: pveril-at-uth.gr
Name: Berillis Panagiotis

Organization: University of Thessaly

Title-Subject: [Filtered] CRT monitor of a Cambridge Stereoscan 240

Message: Dear all

In my lab we have a Cambridge Stereoscan 240 SEM. It is in a working
condition. The CRT monitor of it is really old and deals some problems
(sometimes it appears with a pale light, not the usual rasters that
they appeared when it starts-I think it is a static electricity
problem). Is there any way to put a newer monitor in the SEM? Also I
will be grateful if someone tells me a way to have digital images from
the Cambridge Stereoscan 240.

Regards

Panagiotis

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Email: tina.williams-at-ars.usda.gov
Name: Tina Williams

Organization: USDA

Title-Subject: [Filtered] Zeiss EM9 TEM needs filament change: Help is
now on the way

Message: Thank you all for your responses/suggestions to our request to
find a contact to find/change the filament on a Zeiss EM9 TEM. Help is
on the way.

Thanks,
Tina

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Dear listers,

I recently used our field emission SEM to take X-ray spectrum images, using a 40-square silicon drift detector, of a quite radioactive piece of silicon carbide. What I found interesting was that the Si X-ray maps, as well as the matrix factor in the multivariate statistical analysis (PCA, MCR) decompositions of the spectrum image, looked "streaky" in the fast-scan direction. The features were visible, but it looked like I had printed the images with an inkjet printer with a bad head.

I then loaded an unirradiated specimen of the same material and mapped at the same conditions, and saw no streaking.

Details: JEOL J6500F SEM, 5 kV, 2 nA, ~6000 counts/sec, 10% dead time. 128x100 pix map with 500 micosec/pix. EDAX Genesis v5.31 with EDAX Apollo 40 SDD, 6.4 microsec amp time. With the beam off, the sample gave ~200-300 cps and read 50% dead time (really -- not sure why). The sample was ~1 rem/hr on contact and ~4 mrem/hr at 30 cm.

Any thoughts on what might have been going on? Perhaps the gammas off the specimen were affecting the charge collection? I am at a loss to explain.

Thanks,
Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

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Listers,

Please let me make a clarification. The 300 cps is clearly from the specimen's activity.

What I meant "(really -- not sure why)" about was the lower count rate (300 beam off vs. 6000 beam on) gave a higher dead time (50 vs. 9%).

Apologies,
Chad

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Dear listers,

I recently used our field emission SEM to take X-ray spectrum images, using a 40-square silicon drift detector, of a quite radioactive piece of silicon carbide. What I found interesting was that the Si X-ray maps, as well as the matrix factor in the multivariate statistical analysis (PCA, MCR) decompositions of the spectrum image, looked "streaky" in the fast-scan direction. The features were visible, but it looked like I had printed the images with an inkjet printer with a bad head.

I then loaded an unirradiated specimen of the same material and mapped at the same conditions, and saw no streaking.

Details: JEOL J6500F SEM, 5 kV, 2 nA, ~6000 counts/sec, 10% dead time. 128x100 pix map with 500 micosec/pix. EDAX Genesis v5.31 with EDAX Apollo 40 SDD, 6.4 microsec amp time. With the beam off, the sample gave ~200-300 cps and read 50% dead time (really -- not sure why). The sample was ~1 rem/hr on contact and ~4 mrem/hr at 30 cm.

Any thoughts on what might have been going on? Perhaps the gammas off the specimen were affecting the charge collection? I am at a loss to explain.

Thanks,
Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

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Hi Chad,

your sample was pretty hot! I guess there would have been a lot of
background in the spectrum, perhaps with a big bump around the silicon
peak - I don't know what kind of window your detector has though. 50%
dead time with only the sample itself suggests lots of high energy gamma
rays, but with the beam on the dead time drops to 10%.

Sorry no explanation but your detector is going to see charge being
collected occasionally in large chunks, does the Amp Time change
automatically in your set-up? If the amp-time changes often that might
affect the output at the kind of speed you suggest (horizontal streaks).

Can you hold the amp-time at a fast level rather than allow it to switch?

Good luck
Rob

On 6/21/2013 9:16 AM, parishcm-at-ornl.gov wrote:

Dear listers,

I recently used our field emission SEM to take X-ray spectrum images, using a 40-square silicon drift detector, of a quite radioactive piece of silicon carbide. What I found interesting was that the Si X-ray maps, as well as the matrix factor in the multivariate statistical analysis (PCA, MCR) decompositions of the spectrum image, looked "streaky" in the fast-scan direction. The features were visible, but it looked like I had printed the images with an inkjet printer with a bad head.

I then loaded an unirradiated specimen of the same material and mapped at the same conditions, and saw no streaking.

Details: JEOL J6500F SEM, 5 kV, 2 nA, ~6000 counts/sec, 10% dead time. 128x100 pix map with 500 micosec/pix. EDAX Genesis v5.31 with EDAX Apollo 40 SDD, 6.4 microsec amp time. With the beam off, the sample gave ~200-300 cps and read 50% dead time (really -- not sure why). The sample was ~1 rem/hr on contact and ~4 mrem/hr at 30 cm.

Any thoughts on what might have been going on? Perhaps the gammas off the specimen were affecting the charge collection? I am at a loss to explain.

Thanks,
Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email:parishcm-at-ornl.gov

--
Robert Keyse, DPhil
Research Scientist
Lehigh University

5, East Packer Avenue,
Whitaker Laboratory
Bethlehem,
PA 18015-3194

Office (610)758-3465
Fax (610)758-4244

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Hi Chad,

Is it possible to send the spectrum? I may be able to get a guess of what's happening from the change in shape of the peaks or background.

Also, it may be dependent on what kind of radiation. If you have betas or alphas as well as gammas, I can speculate that they are making it through the charged particle filter in front of your EDS. EDAX makes their filter for 30 keV electrons, not MeV particles. So, if you have a thin window, I could imagine something like an alpha hitting the window material, and initiating a cascade of photons which would be making a large noise background for the detector.

You can also try doing the experiment with a couple filters in front of the EDS. A thin metallized mylar sheet wouldn't hold back gammas, but would block alphas and betas, and would be opaque so you wouldn't get photons through.

Also, do you see any gamma peaks in the EDS spectrum if you acquire a long spectrum with no beam? You can test if the FWHM of the peaks is as expected for your amp time.

Finally, from an electronics level, streaking would be generated by charge not being fully drained and therefore leaking over into the next amp interval. So, it is a pretty sure bet that whatever is going on, a lot of charge pairs are being generated in your silicon, or possibly charge is being directly deposited, I suppose in the case of high energy betas depending on your detector design.

Cheers,

Zack Gainsforth
Space Sciences Laboratory, UC Berkeley
7 Gauss Way
Berkeley, CA 94720
work: 510-642-9733
cell: 626-437-9186
zackg-at-ssl.berkeley.edu

On Jun 21, 2013, at 7:22 AM, parishcm-at-ornl.gov wrote:

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Listers,

Please let me make a clarification. The 300 cps is clearly from the specimen's activity.

What I meant "(really -- not sure why)" about was the lower count rate (300 beam off vs. 6000 beam on) gave a higher dead time (50 vs. 9%).

Apologies,
Chad

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Dear listers,

I recently used our field emission SEM to take X-ray spectrum images, using a 40-square silicon drift detector, of a quite radioactive piece of silicon carbide. What I found interesting was that the Si X-ray maps, as well as the matrix factor in the multivariate statistical analysis (PCA, MCR) decompositions of the spectrum image, looked "streaky" in the fast-scan direction. The features were visible, but it looked like I had printed the images with an inkjet printer with a bad head.

I then loaded an unirradiated specimen of the same material and mapped at the same conditions, and saw no streaking.

Details: JEOL J6500F SEM, 5 kV, 2 nA, ~6000 counts/sec, 10% dead time. 128x100 pix map with 500 micosec/pix. EDAX Genesis v5.31 with EDAX Apollo 40 SDD, 6.4 microsec amp time. With the beam off, the sample gave ~200-300 cps and read 50% dead time (really -- not sure why). The sample was ~1 rem/hr on contact and ~4 mrem/hr at 30 cm.

Any thoughts on what might have been going on? Perhaps the gammas off the specimen were affecting the charge collection? I am at a loss to explain.

Thanks,
Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

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As you are filling out your forms to take advantage of the $100 savings
on registration for the 2013 M&M meetings, please remember that student
bursaries and volunteers are needed to help the meeting run smoothly.
They provide support in the different symposia (help with audio-visual
needs, maintaining an attendance count, and help speakers with set up
for their presentation), staff volunteer office, monitor use of the
Internet Caf�, and help with vendor tutorials.

Student bursaries will be paid $10/hour for assisting with any of the
tasks mentioned above (paid by check at the end of the meetings). There
is an added bonus of $10 cash for each morning and/or afternoon session
worked to help with meals. Volunteers are also eligible for the $10
cash for meals. Both groups get a meeting shirt.
If anyone has any questions about the bursary/volunteer program, or
would like to participate, please contact:

Amanda Lawrence
Institute for Imaging and Analytical Technologies
Mississippi State University
662-325-3019
alawrence-at-i2at.msstate.edu

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Hey listers,

I was wondering if anyone has used hydrid detectors in the NDD position
on a multi-photon microscope, and if so, what they thought of them.

Right now we have PMT NDDs installed on our Leica SP8, but we are
considering upgrading to HyD detectors. We have a couple of HyDs in the
descan position, and they are far more sensitive than PMTs, but they
also can shut-off when the fluorescent signal is too bright. Normally
this is not an issue, but for large tiling runs it can be prohibitively
time consuming to check the gain on the detector for the entire imaging
volume, and yet it is equally frustrating to have a detector quit 3/4 of
the way into an hour long tiling run due to an unexpectedly bright cell,
or worse yet to have a detector quit during live imaging because a cell
became increasingly bright.

We would love the increased sensitivity of HyD NDDs, especially for live
imaging and autofluorescence detection, but I'm not sure if the tendency
of the HyD detectors to shut-off makes HyD NDDs more of a hassle then
they're worth.

Thanks for any feedback,
Ben Smith

--
Benjamin E. Smith, Ph.D.
Samuel Roberts Noble Microscopy Laboratory
Research Scientist II
University of Oklahoma
Norman, OK 73019
E-mail: benjamin.smith-at-ou.edu
Voice 405-325-4391
FAX 405-325-7619
http://www.microscopy.ou.edu/

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Hello,

Our Gatan double tilt holder for our EM420 TEM was recently damaged. Does
anyone on the listserv has a spare double tilt holder they are willing to
part with? The goniometer on the EM420 TEM is common with the other 400
series TEMs, as well as the CM10-CM30 series of TEMs, so holders which fit
these TEMs will work.

Other holders which are compatible with our TEM, particularly
multi-specimen holders, are also of interest. Please contact offline to
discuss terms.

Thank you,
Chris

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I have an old JEOL 840A with a serious vacuum problem. I would like to be able to pump on the system with only a rough pump for an extended period for troubleshooting purposes. I know how to manually control the various pumps and valves, but the SEM still shuts off after 20 minutes if the diffusion pumps are not hot or if the internal pressure is too high. Is there a way to disable the auto shutoff so I can pump with just the rough pump for a few hours or even overnight?

Thank you,
Dave

____________________________
David J. Wilbur, Ph.D.
Instrumentation Specialist
Department of Chemistry
Tufts University
62 Talbot Ave.
Medford, MA 02155
voice:�� 617-627-2163
Fax:�� 617-627-3443
email:�� david.wilbur-at-tufts.edu

==============================Original Headers==============================
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6, 29 -- Subject: SEM Can I Disable auto shut off of JEOL 840A
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Dave,
is it a diffusion pumped SEM or turbomolecular pump fitted?
I don`t know how to override the automatics but you can be sure that the problem is in the pre-vacuum lines somewhere (or the
cooling water / air pressure). It should be easy to find. Open up all the fittings and check for broken o-rings.
One time a had the same problem and it turned out to be a porous o-ring at the low-vac outlet of the turbomolecular pump, so that
in the end the instrument did not reach the trip-level (0.1 Torr) to switch the valves to high-vac conditions.
If you need PDFs write off-line.

Best,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 25.06.13 19:42, schrieb David.Wilbur-at-tufts.edu:
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} I have an old JEOL 840A with a serious vacuum problem. I would like to be able to pump on the system with only a rough pump for an extended period for troubleshooting purposes. I know how to manually control the various pumps and valves, but the SEM still shuts off after 20 minutes if the diffusion pumps are not hot or if the internal pressure is too high. Is there a way to disable the auto shutoff so I can pump with just the rough pump for a few hours or even overnight?
}
} Thank you,
} Dave
}
} ____________________________
} David J. Wilbur, Ph.D.
} Instrumentation Specialist
} Department of Chemistry
} Tufts University
} 62 Talbot Ave.
} Medford, MA 02155
} voice:�� 617-627-2163
} Fax:�� 617-627-3443
} email:�� david.wilbur-at-tufts.edu
}
}
}
}
} ==============================Original Headers==============================
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} 6, 29 -- Subject: SEM Can I Disable auto shut off of JEOL 840A
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Dave:

You need to be very careful about pumping on an SEM using only the
rough pump if it is an oil sealed rotary vane pump. The reason is
that these pumps can usually produce pressures well below the range
of viscous flow (i.e. below about 10 Pa (0.1 Torr)). Under
conditions of viscous flow backstreaming of oil vapors from the
roughing pump cannot occur; however, below the range of viscous flow
backstreaming becomes possible. Leaving a system standing for a long
time while being pumped on only by an oil-sealed rotary vane pump can
produce pressures in it that are low enough to allow it to become
badly contaminated by oil vapors.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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For those of you who have an Edwards S150B sputter coater and needed to replace the HT cable , I have found out the part number for it:

CP25K Gauge head extensions lead 5 m
Ref: D368-13-005

It is actually a lead from a penning gauge that Edwards cut and adapted for the sputter coater, that it why that coax socket was impossible to find in electronics stores. They only used it on this model.
After months of search, we were lucky enough to get the contact of someone who was there when the S150B was built.

Virginia

Virginia Soares
INESC-MN (Microsystems and Nanotechnologies)
R. Alves Redol, 9
1000-029 LISBON
Tel: +351213100300
Fax: +351213145843
www.inesc-mn.pt
www.in-nano.net

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Greetings,
MSU*s Institute for Imaging & Analytical Technologies, is expanding
into new construction in the research park adjacent to campus within the
next 12-18 months. We are looking to this community for lessons learned
regarding the physical move of major research instrumentation (E.g. EM,
CLSM, XRD). For example, is it best to hire a third party to organize
the move (arrange for the pack, move, install)? Or is it best to work
with individual instrument manufacturers for their de-install/re-install
as well as coordinating the company to transport? What worked well and
what would you *do different next time*? We welcome any
advice/suggestions you may have. Please feel free to respond here or
off-line to Amanda Lawrence (alawrence-at-i2at.msstate.edu).

Giselle Thibaudeau, PhD
Director Institute for Imaging & Analytical Technologies
Mississippi State University
phone 662-325-3017

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It probably doesn't matter a great deal what type of oil you use in
an oil-sealed rotary vane pump, backstreaming of organic material
into the system can still occur if pressure in the pumping line drops
below the range of viscous flow (about 10 Pa or 0.01 Torr). The
problem is that the rotor moves at very high speed around inside the
pumping chamber of the pump, and there is very little clearance
between the two. This causes frictional heating and localized
microscopic shear to occur. which, in turn, can break the molecules
of the pump oil down into smaller molecular fragments. These
fragments are more volatile than the pump oil itself, and ultimately
can become the principle source of oil contamination. Installing a
liquid nitrogen trap into the rough pumping can help minimize the
problem if the trap is properly managed (see page 143 of Vacuum
Methods in Electron Microscopy by Wilbur C. Bigelow, Portland Press,
1994 ISBN 1 85578 052 6); otherwise, don't go below the range of
viscous flow.
--
Wilbur C. Bigelow, Professor Emeritus
Materials Sci. & Engr., Univ. of Michigan
Ann Arbor, Michigan 48109-2136
e-mail: bigelow-at-umich.edu;
Fx:734-763-4788; Ph:734-975-0858
Address mail to: 2911 Whittier Court
Ann Arbor, MI 48104-6731

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--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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***************************************************************************************
Forwarded from "Ask a Microscopist"
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realname - Lee Cohen-Gould
Email - lcgould-at-med.cornell.edu
ORGANIZATION - Weill Cornell Medical College
EDUCATION - Graduate College
LOCATION - New York, NY USA
SUBJECT_OF_QUESTION - looking for acrolein
QUESTION - Hi Guys!
A colleague of mine here at Weill Cornell is far braver than I and uses
acrolein to perfusion-fix rodent brains. Apparently, there is a
manufacturing snafu and it is back-ordered until kingdom come. Does
anyone out there have any to spare? I'll go out on a limb here and say
that she'll pay the shipping. You can reply off-list so that we don't
clog Nestor's server with mundane communications.
Thanks much! Hope to see many of you in Indy!
Lee
Mt. Pleasant, MI 48859
(989) 774-3576

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Who is using all the acrolein?
Making tear gas is the main bulk use that I know of.

Very nasty stuff at 100% - spilled a few ml on my lab coat back in 1972 or
73 when the bottle tipped over under the hood. Remember it well for I
thought I'd die. Not breathing for a long time is a very bad feeling.
Sorry I no longer have any to share.

I used it in a fixative mixture for rabbit zygotes along with
glutaraldehyde, paraformaldehyde and a bit of DMSO. It worked great in a
project that did not fix well with our usual fixatives!

Last I bought was from Kodak.

Also hoping to see many in IN.

Pat

Patricia S. Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E � Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-nhlbi.nih.gov
Opinions and experiences related are those of Pat Connelly and
do not represent the NIH.
This message is not confidential and can be freely shared and reproduced.

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I've been struggling with traceable standards for calibrating the TEM. I'm not interested in Mag-i-cal for several reason, cost included. I found online a technical paper from the Yucca Mountain Project (1989) that indicates if you use a NIST standard to calibrate your SEM and then slide the SEM standard out of the beam and a TEM replica grating into the beam you can check the replica grating and establish NIST traceability. Here's the link.

http://pbadupws.nrc.gov/docs/ML0316/ML031680677.pdf

Any thoughts or comments on this procedure? If this does provide true NIST traceability it could solve my A2LA problems. Maybe yours too.

I'm working toward the US 4th of July holiday and I'm reminded of that old brain tease:

Does Canada (or any other country) have a 4th of July?

Yes they do. They just don't celebrate it.

This joke works best if you're an American at fireworks or a parade...Cheers!

Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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12, 24 -- From frank_karl-at-ardl.com Mon Jul 1 07:53:08 2013
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X-from: uwe.breuer-at-fz-juelich.de ()

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Email: uwe.breuer-at-fz-juelich.de
Name: Uwe Breuer

Organization: Research Centre Juelich, Germany

Title-Subject: [Filtered] Position for Operating a 3D Atomprobe/
SEM-FIB/ SIMS

Message: Hallo,
we will get a CAMECA LEAP 4000X System in September and we are looking
know for someone who has experience in sample preparation using FIB or
with the Atomprobe itself. More detailed description you find here:

http://www.fz-juelich.de/SharedDocs/Stellenangebote/_common/dna/2013-128-EN-ZEA-3.html

Kind Regards

Uwe Breuer

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Group,

Trying to locate parts for a Leo 906E TEM sample holder.

Any suggestions would be helpful.

Roy Beavers
Southern Methodist University
Department of Earth Sciences
3225 Daniel Ave
Dallas, TX� 75205
Voice: 214-768-2756
Email: rbeavers-at-smu.edu

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Frank Karl asked about calibration of a TEM replica grating in an SEM. The
method he described--first measuring a traceable standard and then measuring
another object--is perfectly acceptable. In this method, the traceable
standard acts as a transfer standard because its calibration is transferred
to the microscope and then to the test grating. We use this method
ourselves.
We have published several papers jointly with scientists at various national
metrology labs, such as NIST in the US, PTB in Germany, and NMC in
Singapore. In one recent paper, our 70-nm pitch standard was measured in
our lab using a general purpose AFM with the transfer standard method and at
NMC and NIST using purpose-built AFMs where the length scales were directly
traceable to the SI meter using interferometry. The mean pitch values
obtained by the 3 labs agreed within .025 nm. See
"Multilaboratory comparison of traceable atomic force microscope
measurements of a 70 nm grating pitch standard", Ronald Dixson, Donald A.
Chernoff, Shihua Wang, Theodore V. Vorburger, Siew Leng Tan, Ndubuisi Orji,
Joseph Fu, J. Micro/Nanolith. MEMS MOEMS 10, 013015 (Mar 08, 2011);
doi:10.1117/1.3549914 . Online Publication Date: Mar 08, 2011

In order to achieve traceability, you need both a mean value and an
uncertainty of the mean value. Multiple measurements, statistical analysis
and consideration of various instrumental and sample-related factors are
required. You may also find it useful to know the variability of individual
pitch measurements, so that you can estimate the magnification uncertainty
of images that contain just one or two pitch intervals. This requires even
more measurements.

There is significant work involved with all this and we can help. We sell
software for pitch analysis (DiscTrack Plus) and we provide a commercial
traceable calibration service. Please contact me offline for more details.
regards,
Don
=============================================
Don Chernoff, Ph.D., President
Advanced Surface Microscopy, Inc.; 3250 N. Post Rd., Suite 120;
Indianapolis IN 46226 USA
E-Mail: donc-at-asmicro.com www.asmicro.com
Voice: 317-895-5630 Toll free: 800-374-8557 (in USA & Canada)
Fax: 317-895-5652
[business activities since 1990: analytical services in AFM, AFM probes,
consulting, training, calibration and test specimens,
calibration and measurement software, used NanoScope equipment.]
=============================================

----- Original Message -----
From: frank_karl-at-ardl.com
To: donc-at-asmicro.com
Sent: Monday, July 01, 2013 8:57 AM
Subject: [a] [Microscopy] Once again, into the TEM calibration breech

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I've been struggling with traceable standards for calibrating the TEM.
I'm not interested in Mag-i-cal for several reason, cost included. I found
online a technical paper from the Yucca Mountain Project (1989) that
indicates if you use a NIST standard to calibrate your SEM and then slide
the SEM standard out of the beam and a TEM replica grating into the beam you
can check the replica grating and establish NIST traceability. Here's the
link.

http://pbadupws.nrc.gov/docs/ML0316/ML031680677.pdf

Any thoughts or comments on this procedure? If this does provide true
NIST traceability it could solve my A2LA problems. Maybe yours too.

I'm working toward the US 4th of July holiday and I'm reminded of that old
brain tease:

Does Canada (or any other country) have a 4th of July?

Yes they do. They just don't celebrate it.

This joke works best if you're an American at fireworks or a
parade...Cheers!

Frank

This email and any of its attachments may contain confidential information
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received this email in error, please notify us by return email at
info-at-ardl.com, permanently delete the email, and destroy any printouts. If
this email contains test data and/or draft reports, you are hereby notified
that only a signed original test report will contain official results, a
copy of which resides in the project folder located at ARDL, Inc. Thank you.
Akron Rubber Development Laboratory, Inc.

==============================Original
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Yes, this to think about as we prepare to travel to Indianapolis. In 1897, the Indiana House passed, and the Senate nearly passed a bill that attempted to define the area of a circle using a bizarre formula for squaring a circle, which would have required pi to have a value of 3.2 instead of 3.14159265. I am not making this up. You can read more about in the Wikipedia article at:

http://en.wikipedia.org/wiki/Indiana_Pi_Bill

John Mardinly, ASU

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Hello John,
I seem to remember other communities wanting to change the value of Pi to make it easier for school children to remember.

If Indiana had passed this law one has to wonders if the state police would have been stopping cars on the highway to check if the wheels were regular polygons. I see it now..."Off to jail with you, you round scofflaw!

Frank

-----Original Message-----
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Sent: Tuesday, July 02, 2013 5:38 PM
To: Frank Karl

Yes, this to think about as we prepare to travel to Indianapolis. In 1897, the Indiana House passed, and the Senate nearly passed a bill that attempted to define the area of a circle using a bizarre formula for squaring a circle, which would have required pi to have a value of 3.2 instead of 3.14159265. I am not making this up. You can read more about in the Wikipedia article at:

http://en.wikipedia.org/wiki/Indiana_Pi_Bill

John Mardinly, ASU

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Title-Subject: [Filtered] Wire saw

Message: Hi All

Can anyone recommend a good wire saw vendor? Please contact me offline: mlibbee-at-gmail.com or
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Organization: Ghent University, Department of Morphology

Title-Subject: [Filtered] SEM on eggs of seabass and sole

Message: Dear colleagues,

we want to do SEM analysis on eggs of seabass and sole. For seabass, this is no problem, but when we
apply same protocol on sole eggs, these eggs will shrink as soon as the dehydration protocol starts.

This is our protocol:
fixation in HEPES with 2% paraformaldehyde en 2,5% glutaraldehyde for several weeks, postfixation in
1% OsO4 for 2 hours. Then, we start the dehydration process in 50% alcohol up to 90%, finally
aceton, followed by CPD and platinum sputtering.

I really hope that some of you can give me some advice.

Kind regards,
Wim.

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Dear Wim,

You could try something that works on watery plant cells: Fix in methanol.
It works really well and gives the least shrinkage. Ref:

Neinhuis C, Edelmann HG: Methanol as a rapid fixative for the
investigation of plant surfaces by SEM. Journal of Microscopy 1996,
184(1):14-16

http://onlinelibrary.wiley.com/doi/10.1046/j.1365-2818.1996.d01-110.x/abstr
act

Modified as follows:
Leaf pieces were immersed in 100% dry methanol for 10 min, followed by 2 x
10 min changes in 100% dry methanol. Tissue was then critical point dried
immediately with methanol as the transitional fluid.

Alternatively, tissue can be transferred into fresh dry ethanol and left
overnight at 4�C (essential for larger tissue pieces which retain water
for longer). Tissue pieces were then critical point dried with ethanol.

Essential is CPD straight away, no storage in solvent.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Faculty position in Structural Biotechnology/CryoEM

An assistant professor position is available at the EM facility affiliated with KTH Royal Institute of Technology and Karolinska Institutet in Stockholm. The laboratory is equipped for a wide variety of applications of cryoEM. It is located in a vibrant surrounding focusing on basic biomedical research, including an infrastructure for advanced light microscopy, as well as medical research at the university hospital and medical engineering at KTH.
General assessment criteria include ability to establish and develop independent research and initiate collaborations in research and education. Applicants should have outstanding experience of cryoEM methodology or be able to demonstrate such a strong background in another technique of structural biology that it appears likely that expertise in electron microscopy can be acquired.

The position is associated with the academic career path at KTH. http://www.kth.se/en/om/work-at-kth/tenure-track/tenure-track-den-akademiska-karriarvagen-1.55796

For further details including information of the application procedure, see:
http://www.kth.se/en/om/work-at-kth/vacancies/assistant-professor-in-structural-biotechnology-1.398165

or contact:

Hans Hebert
Dean, School of Technology and Health
KTH Royal Institute of Technology
Alfred Nobels All� 10
S-141 52 Huddinge, Sweden
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Title-Subject: [Filtered] FOM-FIG Lunch With A Manager-MM 2013

Message: Greetings,

Second Notice: Lunch With a Manager.

Roundtable discussion with a panel of veteran Facility Managers, Supervisors, and Directors
[Academia & Industry] aimed at future Microscopy Professionals (students, postdocs, etc.). If you
know a student of other interested person considering a career in Facility Operation and Management,
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Email: albertofabrizi.it-at-gmail.com
Name: Alberto Fabrizi

Organization: Universit� di Padova, Dipartimento di Tecnica e Gestione dei sistemi industriali (DTG)

Title-Subject: [Filtered] TEM JEM 2000 EX - Oil Diffusion Pump heater alarm lamp

Message: Dear colleagues,

we have a TEM JEOL JEM 2000EX II, and three weeks ago, the alarm lamp indicating the breakup of oil
diffusion pump heater was turned on.
We have changed the DP heater with new one and it seems to heat correctly, anyway the microscope
continues to switch off after about 20 minutes from the starting and the DP heater alarm turns on again.

Do you have any experience on such trouble?
In addition to the breakup of the DP heater, are there any other reasons for the DP alarm lamp turn-on?
Could the problem be more related to the poor grade of vacuum reached by the rotary pump?

Thank you in advance for any your help.

Best regards,

Alberto.

Alberto Fabrizi
Universit� di Padova
Dipartimento di Tecnica e Gestione dei sistemi industriali (DTG)
Stradella San Nicola, 3
36100 Vicenza (Italy)

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Email: z.zhou-at-lboro.ac.uk
Name: Zhou

Organization: Loughborough University

Title-Subject: [Filtered] TEM sample prep --- Disc Punch

Message: We need a disc punch for 3mm diameter disc TEM sample preparation. Often Al-, Ni-, and Ti-
alloys are of our interest for twin-jet electro-polishing. Suppliers are Gatan, South-Bay
technology, and SPI supplies. I didnt use any of these disc punches. We have multiusers in a
university lab. I would be very grateful if you would share some of your experience with me.

Zhou

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As I mentioned before, I'm affiliated with a Chicago hackerspace known as
Pumping Station: One. We have acquired two SEMs, a functional Leica S430
and a non-functional S440. This is taking up too much space, and we need
to get rid of the S440. What we know about it:

It has secondary electron, 4 quadrant BSD, and an additional detector that
I believe may be cathodoluminescence. It has a motor controlled stage.
The column appears to have an upper ion pump and gate valves.

There appears to be a problem with the Pentium board. It originally booted
Windows, but froze. Then it would almost get to booting Windows, but it
froze. Then, it would freeze in the BIOS. Then, it wouldn't boot at all.
We purchased a similar model Pentium board to try to replace it. That
didn't work, and we decided to throw in the towel.

Having experience disassembling and reassembling the S430, it appears that
the S440 is complete and in good physical condition. Without being able
to run the Leica software, I can't speculate further.

We are willing to accept any reasonable cash offer for it, or would consider
donating it to a non-profit organization. Otherwise, we anticipate parting
it out.

Regards,
Ryan

==============================Original Headers==============================
6, 15 -- From rpierce-at-elna.mackenziegems.com Mon Jul 8 10:35:51 2013
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6, 15 -- From: Ryan Pierce {rdpierce-at-pobox.com}
6, 15 -- To: Microscopy-at-microscopy.com
6, 15 -- Subject: SEM: Leica S440 in Chicago needs new home
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Dear Alberto,

You may find a thermal detector switch fixed on the DP. If DP heater is
operationg correctly this contact is closed because the DP body is warm.
But this thermal detector itself can be out of order. Be careful because
there is another thermal detector fixed on the cooled area of the DP
body. This one is to detect if there is enought water on the hose or not.
Regards.
Nicolas Stephant

Le 08/07/2013 14:44, microscopylistserver-noreply-at-microscopy.com a �crit :
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} Email: albertofabrizi.it-at-gmail.com
} Name: Alberto Fabrizi
}
} Organization: Universit� di Padova, Dipartimento di Tecnica e Gestione dei sistemi industriali (DTG)
}
} Title-Subject: [Filtered] TEM JEM 2000 EX - Oil Diffusion Pump heater alarm lamp
}
} Message: Dear colleagues,
}
} we have a TEM JEOL JEM 2000EX II, and three weeks ago, the alarm lamp indicating the breakup of oil
} diffusion pump heater was turned on.
} We have changed the DP heater with new one and it seems to heat correctly, anyway the microscope
} continues to switch off after about 20 minutes from the starting and the DP heater alarm turns on again.
}
} Do you have any experience on such trouble?
} In addition to the breakup of the DP heater, are there any other reasons for the DP alarm lamp turn-on?
} Could the problem be more related to the poor grade of vacuum reached by the rotary pump?
}
} Thank you in advance for any your help.
}
} Best regards,
}
} Alberto.
}
} Alberto Fabrizi
} Universit� di Padova
} Dipartimento di Tecnica e Gestione dei sistemi industriali (DTG)
} Stradella San Nicola, 3
} 36100 Vicenza (Italy)
}
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--
Signature courrier �lectronique Nicolas STEPHANT

Universit� de Nantes
Institut Jean Rouxel
Service de microscopie �lectronique � balayage et microanalyse
2 rue de la Houssini�re
BP 92208
44322 Nantes c�dex 3

T�l : 02 40 37 64 26
M�l : nicolas.stephant-at-univ-nantes.fr
Web : http://www.univ-nantes.fr/smebm

"Le monde n'existe que pour autant que nous sommes capables d'en
produire une image"
C.G Jung

==============================Original Headers==============================
7, 26 -- From Nicolas.Stephant-at-univ-nantes.fr Mon Jul 8 11:16:21 2013
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7, 26 -- Subject: Re: [Microscopy] viaWWW:TEM JEM 2000 EX - Oil Diffusion Pump heater
7, 26 -- alarm lamp
7, 26 -- References: {201307081244.r68CiSm3004723-at-ns.microscopy.com}
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We have a Nikon TI with the TIRF arm. For many years, when you turned the microscope on, the system defaulted to Epi mode and if you wanted to do TIRF, you would select TIRF on the RCP box. Recently, the system has been defaulting to TIRF, so users have to remember to switch to Epi on the RCP in order to see widefield fluorescence. Does anyone know how to change the default behavior? Thanks- Dave

David Knecht, Ph.D.
Professor and Head of Microscopy Facility
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

David Knecht, Ph.D.
Professor and Head of Microscopy Facility
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

David Knecht, Ph.D.
Professor and Head of Microscopy Facility
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

==============================Original Headers==============================
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Hello All,

We were hoping for some help on a search we are working on. Perhaps, this opportunity might be right for you or someone you know, who could benefit from speaking with us.

We are executing a search for a person who has experience in the following key areas: Microscopy, TEM, PLM technical knowledge of Asbestos, Geology, Mineralogy and other duties related to Asbestos analysis.

This person could be an experienced laboratory manager or a strong analyst who is looking for career advancement.

We are a confidential search firm with an excellent reputation for such and treat all inquiries accordingly. I can be reached at:

paul at jpamri.com or 866-712-1810 Paul Palazzolo, Senior Managing Partner, JPA

We thank you in advance for your time and consideration

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Does anyone out there know of an XL30 SFEG SEM users group I can join?

Thanks in advance

Fred

Fred Hayes
Manager
Advanced Materials Characterization and Testing Lab (AMCaT)
Dept of Chemical Engineering and Material Science
3001 Ghausi Hall
Univ of CA Davis
Davis, CA 95616
530-752-0284
http://chms.engineering.ucdavis.edu/index.html
http://chms.engineering.ucdavis.edu/research/amcat/index.html

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7, 22 -- To: {Microscopy-at-microscopy.com}
7, 22 -- Subject: XL30 SFEG SEM users group
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Hi Fred,

Not aware of XL30-specific users group, but I am running (and keeping
alive) a tool that was born back in 1995 as XL40 and at the later time
got "converted" into FIB620 dual-beam by addition of FEI FIB column.

During past year had lots of fun with it, including rebuilding it's
oil-filled (would you believe that for in-vacuum design?!???) rotation
stage and just last week finished restoring its main PC from a "clinical
death" caused by a power glitch (CPU and GPIB cards were gone and hard
disk got bad sector right on the ntkernel.exe file)... .

Cheers :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 7/8/2013 9:40 PM, fahayes-at-ucdavis.edu wrote:
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} Does anyone out there know of an XL30 SFEG SEM users group I can join?
}
} Thanks in advance
}
} Fred
}
} Fred Hayes
} Manager
} Advanced Materials Characterization and Testing Lab (AMCaT)
} Dept of Chemical Engineering and Material Science
} 3001 Ghausi Hall
} Univ of CA Davis
} Davis, CA 95616
} 530-752-0284
} http://chms.engineering.ucdavis.edu/index.html
} http://chms.engineering.ucdavis.edu/research/amcat/index.html
}
}
}
}
} ==============================Original Headers==============================
} 7, 22 -- From fahayes-at-ucdavis.edu Mon Jul 8 20:39:28 2013
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} 7, 22 -- To: {Microscopy-at-microscopy.com}
} 7, 22 -- Subject: XL30 SFEG SEM users group
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Email: paul-at-foresight.org
Name: Paul Melnyk

Organization: Foresight Institute

Title-Subject: [Filtered] WANTED: Atomic Force Microscope or Scanning Electron Microscope

Message: We are looking for an Atomic Force or Scanning Electron microscope.

Various states of operation or condition will be considered as long as repair is possible.

Ideally, the equipment would be donated to our 501(c)(3) Not-for-Profit organization.

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Please contact me with any possibilities.

Thank you!

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Paul Melnyk, President
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realname - Stefanie Leland
Email - stefanie.leland-at-gmail.com
ORGANIZATION - Jar of Grasshoppers Productions
EDUCATION - Undergraduate College
LOCATION - Oklahoma City, OK, USA
SUBJECT_OF_QUESTION - Microscopes recommendations
QUESTION - Hello,

I have a Canon 60D and am wanting to start working with microscopic
photography. I've been doing macrophotography for some time, but am
interested in getting a closer look into some of life's amazing
things...insects, bacteria, plants, yeast, etc. I would like to be able
to adapt my camera to a trinocular port in order to photograph my
findings. I haven't used a microscope in many years, but am on the
search for a good microscope at an affordable price. Between $250 and
$400. I realize that the camera adapter is going to be fairly costly as
well. Would you have any recommendations for a camera adapter for my
Canon DSLR? I don't know if I can find what I am looking for in this
price range, but was hoping that I may find some help at this site.

Sincerely,
Stefanie Leland
Jar of Grasshoppers Productions

.

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7, 28 -- From oshel1pe-at-cmich.edu Tue Jul 9 07:37:43 2013
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Hello,

my institute wants to get rid of an old Phillips CM200 TEM.
It will be disassembled and scraped. So there are some spares available.
CCD, EDX and FEG are allready gone.
If you need anything else let me know :-)

Unfortunatly for legal reasons this is an EU-only offer.

Cheers

Max

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Email: kalennon-at-hagerstowncc.edu
Name: Kristen Lennon

Organization: Hagerstown Community College

Title-Subject: [Filtered] confocal wanted

Message: Hi,
Hagerstown Community College in Western Maryland is setting up a microscopy training hub as part of
our biotechnology program. The Hub is focussed on bolstering microscopy skills in our graduates who
often go on to jobs in the biotech industry and in the K-12 STEM community.

As part of this effort, we are looking for a donation of a confocal microscope in good working
condition. We would be able to pay for shipping or arrange for pick up within the area.The donor
would be acknowledged in any materials generated and on our Biotechnology and/or Microscopy Training
Hub web site, the latter of which is in development.

This microscopy program has already impacted over 2500 students in our region since it began in
January 2013, and has great promise to enhance STEM engagement and education and a much greater
level. The current facilities have been supported by a grant from the National Science Foundation
that does not include monies for the purchase of a confocal.

If you have any leads on a confocal, please contact me.

Kristen A. Lennon, Ph.D.
Biotechnology Microscopy Outreach Coordinator
Hagerstown Community College
11400 Robinwood Dr.
Hagerstown, MD 21742

kalennon-at-hagerstowncc.edu

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Hi

I have just tried a optical coupler for DSLR sold as a set with
different adaptation tubes for old and new eyepiece tube diameters and C
thread. You need in addition the T-mount for your DSLR. Type this
reference on the internet and you should find it : MA421101

Its simple to use, and the mounting kit is very universal, but the
optical quality is say just good in the centre of the field, but has a
loss of sharpness in the corners and so some chromatic aberration. If
you cut a smaller rectangle/square in the central part of the picture,
it can be an interesting solution. But it will decreases a step more the
size of the captured field of view.

A second way, is to use a good eyepiece (you have it with the
microscope) and a f50 mm lens from an full frame film SLR. It's a far
cheaper solution, gives more field and a better image quality, but with
the need to feedle something to mount the DSLR on the photo-tube. I made
my comparison tests with a Industar lens (a russian Tessar, very cheap
and very good for it price...).

Hope it helps

Jacques

J. Faerber
Strasbourg - France
E-mail : Jacques.Faerber-at-ipcms.unistra.fr

Le 09/07/2013 14:49, oshel1pe-at-cmich.edu a �crit :
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}
} realname - Stefanie Leland
} Email - stefanie.leland-at-gmail.com
} ORGANIZATION - Jar of Grasshoppers Productions
} EDUCATION - Undergraduate College
} LOCATION - Oklahoma City, OK, USA
} SUBJECT_OF_QUESTION - Microscopes recommendations
} QUESTION - Hello,
}
} I have a Canon 60D and am wanting to start working with microscopic
} photography. I've been doing macrophotography for some time, but am
} interested in getting a closer look into some of life's amazing
} things...insects, bacteria, plants, yeast, etc. I would like to be able
} to adapt my camera to a trinocular port in order to photograph my
} findings. I haven't used a microscope in many years, but am on the
} search for a good microscope at an affordable price. Between $250 and
} $400. I realize that the camera adapter is going to be fairly costly as
} well. Would you have any recommendations for a camera adapter for my
} Canon DSLR? I don't know if I can find what I am looking for in this
} price range, but was hoping that I may find some help at this site.
}
} Sincerely,
} Stefanie Leland
} Jar of Grasshoppers Productions
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I'm looking for some information on freezing point depression of a mixture of methanol, DMSO and water. I anticipate cutting ultra thin sections of H-NBR for TEM examination. While I know Tg changes with acrylonitrile content, degree of branching and degree of hydrogen saturation, none of that information will be available to me and I need a nice clean solvent to float my thin sections. I've tried saturated sugar in the past, but I'm not happy with it.

I can find plenty of binary mixtures with low freezing points, (ie: ethanol and water) but methanol, DMSO and water was suggested. I'm unable to find any data, including ratios and freezing temperature.

Any advice would be welcome.

Thanks.......

Frank
ARDL

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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realname - Henry Rivas
Email - henry.rivas-at-wpxenergy.com
EDUCATION - Graduate College
SUBJECT_OF_QUESTION - TM-3000 or other SEM
QUESTION - Where can I buy a used over the desk SEM like the TM-3000?

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Trying to find someone with experience with this combination(T330 SEM and EMS-SEM Digi-CAM) to replace Polaroid 4 X 5 in sheet film. (Yep!..still trying to solve these problems, but it is turning into the old digital vs. wet photographic "what are we losing?" discussion/study...and I don't give up!)

Jeff in Texas
wa5ekh-at-juno.com
____________________________________________________________
1 Weird Fruit Melts Fat
Cut Pounds of Stomach Fat Every Week With This Secret Fruit
http://thirdpartyoffers.juno.com/TGL3131/51ddddd5d7e7c5dd527e0st04vuc

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I have a client who wants to look at a fungal infection of jute with TEM.
Fixation will be done in a remote location, so cryo is out, and maybe
osmium tetroxide is also out (but he doesn't seem to be fazed by
glutaraldehyde). Plus I'd like to give him a protocol for a resin easier
to prepare than the usual four-part epoxy.

Any suggestions?

Aloha from Honolulu, where mango season is in full swing.
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*
****************************************************************************

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Hello Listers,
If anyone has a functional OXFORD Tetra BSE hand held controller they
would like to part with, I could use one. The switches on mine bad, but
the BSE system does function, only with a fixed gain. Thanks in
advance.

Gary Easton
Scanners Corporation
Third Party SEM Service/IXRF Systems EDS Analyzers
410-857-7633 x102

-----
No virus found in this message.
Checked by AVG - www.avg.com
Version: 2013.0.3349 / Virus Database: 3204/6482 - Release Date: 07/11/13

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Hello all,
Please, could anybody give us an advice or hint on following problem?
After cathode (W) exchange in our Philips CM100, we are not able to get
ultra high vacuum. IGP pump switch off and on repeatedly and also the
valves V3 and V4 are closed and open in the same cycle.
The LED on IGP HT source is lighting all the time.
I have taken two snaps of OPCON VACUUM page. The link for images is:

http://www2.biomed.cas.cz/~benada/IGP_trouble2013.html

Thanking in advance for any response.

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR. v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

==============================Original Headers==============================
5, 37 -- From benada-at-biomed.cas.cz Mon Jul 15 08:15:49 2013
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5, 37 -- From: Oldrich Benada {benada-at-biomed.cas.cz}
5, 37 -- To: Microscopy-at-Microscopy.com
5, 37 -- Subject: Philips CM100 IGP trouble
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Hello Oldrich,
I only have an EM420 with IGP but maybe my experience might help you (if the problem is so easy):

Did you use dry nitrogen and did you flush the gun with it during cathode change?
If not you might have a "wet" IGP now.
Is your IGP (after ca one hour of pumping) getting hot? If yes, let it come up to ca. 60-80�C and switch off only the IGP.
Wait one hour for the IGP to cool, then switch it on again.
IGP high vacuum should recover within ca. 4-6 hours.
Switching on and off is a sign that your scope is just at the trip level between diffusion pump high vac and IGP high vac
condition (at least that`s the case at the EM420).

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 15.07.13 15:21, schrieb benada-at-biomed.cas.cz:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
}
} Hello all,
} Please, could anybody give us an advice or hint on following problem?
} After cathode (W) exchange in our Philips CM100, we are not able to get
} ultra high vacuum. IGP pump switch off and on repeatedly and also the
} valves V3 and V4 are closed and open in the same cycle.
} The LED on IGP HT source is lighting all the time.
} I have taken two snaps of OPCON VACUUM page. The link for images is:
}
} http://www2.biomed.cas.cz/~benada/IGP_trouble2013.html
}
} Thanking in advance for any response.
}
} Best regards Oldrich
}
} --
} Oldrich Benada
} Institute of Microbiology AS CR. v.v.i.
} Videnska 1083
} 142 20 Prague 4
} Czech Republic
}
} ==============================Original Headers==============================
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} 5, 37 -- To: Microscopy-at-Microscopy.com
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11, 23 -- From stefan.diller-at-t-online.de Mon Jul 15 08:41:51 2013
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Dear All,
I got a Spicer SC12 field cancelling system without the cables.
Since I am thinking of setting up a Helmholtz cage around my SEM column I would like to know if anybody out there might help me
with the coils, like what kind of coils and diameter of the wires, impedance,...
I would also be very grateful to get a complete user manual.

Best wishes,
Stefan

--

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

==============================Original Headers==============================
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The Department of Physiology and Neurobiology at the University of Connecticut, Storrs seeks applications for an Academic Assistant I/II in the Electron Microscopy Laboratory. The successful candidate will assist other laboratory staff in training and supervision of laboratory users, maintenance of equipment, sample preparation and general laboratory maintenance. This is a full-time, annually renewable position.

Go to www.jobs.uconn.edu and click on Staff Openings } Job ID 2014007 (Academic Assistant I/II) for more information. Screening of applications will begin immediately and will continue until the position is filled. Preference will be given to applications received before July 31, 2013. We encourage applications from under-represented groups, including minorities, women, and people with disabilities. The University of Connecticut is an EEO/AA employer.

Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369

==============================Original Headers==============================
5, 17 -- From marie.cantino-at-uconn.edu Mon Jul 15 11:58:54 2013
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Dear Listers,

There is a researcher in Austin, Texas who needs TEM service to look at some plant tissue. Since I'm a member of MSA I offered to put this request out on the Listserver. There has to be someone out there closer than me (I'm in Omaha, NE). So hopefully, someone down that way will see this request, if you are willing to help please contact me.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

The information in this e-mail may be privileged and confidential, intended only for the use of the addressee(s) above. Any unauthorized use or disclosure of this information is prohibited. If you have received this e-mail by mistake, please delete it and immediately contact the sender.

==============================Original Headers==============================
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Email: anwar-at-kfupm.edu.sa
Name: Dr. Anwar Ul-Hamid

Organization: King Fahd University of Petroleum & Minerals

Title-Subject: [Filtered] Vacancy Announcement

Message: Vacant Position
Research Scientist - Transmission Electron Microscopy

Job Description
Candidate will be proficient in the use of field emission transmission electron microscopy with
associated techniques such as microchemical analysis, sample preparation, image interpretation,
crystallography, etc using latest available techniques. Candidate will conduct basic and applied
research on characterization of metals, alloys, ceramics and polymeric materials. He will apply
knowledge of transmission electron microscopy to understand the imaging, structure and
microchemistry of a wide range of materials and document research results. He will participate in
research programs, work for funded projects and provide analytical services to students and
researchers at the university. He will apply advanced characterization tools to study the
microstructure, composition, and properties associated with materials. His experience will ensure
that he does not require any training in transmission electron microscopy. He will maintain high
quality data standards and will have knowledge of operation of a TEM lab to aim for high
productivity in a high work volume setting. Candidate will work as member of a team and is expected
to develop good communication and working relationship with the university scientific community. He
is expected to handle multiple projects and stay on schedule for project deliverables. He will
maintain and monitor equipment. He is expected to invent, patent, present and publish wherever
appropriate.

Eligibility
MS, preferably PhD degree in Materials Science or closely related field with at least 3 years
experience in advanced transmission electron microscopy techniques including sample preparation,
focused ion beam, image interpretation, microchemical analysis, high resolution imaging, electron
energy loss spectroscopy and electron diffraction structural analysis.

Contact:
Dr. Anwar Ul-Hamid
King Fahd University of Petroleum & Minerals
Dhahran 31261
Saudi Arabia
anwar-at-kfupm.edu.sa

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Anyone have experience with a GW Type 30A Quad Backscatter?? Forgot how to run it and there is no manual.

Note: Still trying to find a way to take micrographs with a surplus JEOL T33O A on a very low budget (to prove it works well enough to spend more money next semester/physical year).

Jeff Day
wa5ekh-at-juno.com
____________________________________________________________
New Policy in Mississippi
2012-Drivers w/ no DUIs eligible for up to 50% off car insurance...
http://thirdpartyoffers.juno.com/TGL3131/51e87d90492cb7d904fd8st04vuc

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Email: david.tanner-at-ul.ie
Name: David Tanner

Organization: University of Limerick

Title-Subject: [Filtered] App for learning TEM?

Message: Hi,

I was wondering if anybody has come across an app to help students learn how to use TEM?
When lecturing the basics of TEM, I always refer students to the Matter web site which gives an
excellent overview of how a TEM works (http://www.matter.org.uk/tem/). I wondered if anybody knows
of an app similar to this that students could take with them on their iPad or iPhone when using the
TEM to help them develop their knowledge?
Maybe the electron microscopy companies might have apps for aligning / using a particular instrument
...?

Thanks
David.

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Does anyone have experience with an L.C error on JEOL JSM-5800 SEMs or
similar JEOL units? I recently purchased a used 5800, which functioned
prior to moving. After moving, the unit starts up fine up to the HT
ready point. When HT is pressed the L.C error appears. This happens
whether or not filament current is applied. The filament has been
replace and the Wehnelt cap has been cleaned. As a test, I removed the
filament and still get the same error.
The only reference to this error in the operating manual is under poor
image quality troubleshooting. The manual list the error as "flucture
load current (L.C)" and suggests cleaning the Wehnelt cap or replacing
the filament. Currently no image can be obtained.
I am thinking that something in the HT tank was damaged during the
move. Does anyone have a drawing or schematic for the tank? Does
anyone have a service manual for the JSM-5800 or similar unit? I am
also open to troubleshooting suggestions and suggestions for repair.

Thanks,
Fred Anderson

--

Fred Anderson, P.E.
Advantec Engineering LLC

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Hi

The problem is likely to be in the high voltage tank or the cable. To check
out the cable remove the connection from the tank end (cover with foil) and
check the LC again. Problem has gone then it's your cable which any
organisation working with high voltage (x-rays) should be able to help you
at a much lower cost than the manufacturer.

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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Sent: 20 July 2013 14:32
To: protrain-at-emcourses.com

I'm completely confused by the numbers I'm getting from my S430. My understanding is that beam current measures electrons in the column, and probe current measures electrons striking the sample. Probe current is less than beam current because things like apertures prevent all the electrons in the column from striking the sample.

Now I would think that for a given column configuration, probe current and beam current have to increase or decrease together. I'm not seeing that, and I don't understand it.

I've set up a data zone with both probe current and beam current displayed. When I increase or decrease the probe current, I see the secondary electron image get lighter or darker, as I expect, but beam current stays the same.

Adjusting beam current isn't intuitive. I have to open a status box and add it as a data field, but then I can type it in. It seams the maximum beam current is 400 uA. When I change the value, the image gets darker or lighter as I expect, but the probe current doesn't change!

To make matters more interesting, I can change the filament current, and it makes the image lighter or darker. I imagine as the temperature of the filament changes, its thermionic emission changes, causing more or less electrons to be released, hence a brighter or darker secondary electron image. But wouldn't that change both beam and probe current? I change this and both beam and probe current as reported by the S430 stay the same.

I'm completely confused! These numbers on the S430 don't seem to match what I expect.

Also, my understanding is that one can measure probe current by shooting the beam into a faraday cup and measuring the resulting current as it leaves the stage. While I don't have one of these yet, I do see the ground for the stage coming through a feedthrough in the front door. What kind of connector is it, and where can I find one? It turns out the hackerspace has a Keithley electrometer that can measure picoamps, and I'm guessing I can hook this inline somehow. (That takes a BNC triax connector, which I'd need to find.

Any thoughts?

Regards,
Ryan

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Dear All, may i know if you
have any advices for cleaning the samples that are supported by the
carbon films on the copper grids? I did find some advices from the past
emails in the mailing list. Any comments on using H2 /O2 gases as what
is implemented in Gatan Solarus? As i am using Fishione Nanoclean, i am
thinking if getting H2 gas with O2 gas will yield the same result. Due
to the safety reasons in lab, the safety officer suggested that i get a
H2 generator rather than H2 gas cylinder, but it does cost quite an
investment. I would like to thank Fishione , which also kindly provide
me some recipe on this cleaning too (but not based on H2/O2). On the
other hand, though, I am hoping if someone could share some advices on
this too?

Cheers,
Yee Yan, Tay
FACTS LabNTU, Singapore

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6, 40 -- From: Tay Yee Yan {one_twinklestar-at-yahoo.com.sg}
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I think you may want to check the definitions. Probe current should be on the order of a nA. Beam current could be the filament heating current. I don't know the S430, so I cannot say. Changing the "beam current" could be changing saturation and should lead to some change in probe current but it would not be a direct change. It depends where you are on the saturation curve.

Changing probe current is adjusting the condenser lens and is totally separate from what is happening in the gun.

There would also be specimen current - that which is absorbed by the sample. That will depend on the sample and could vary for the same probe current.

But like I said, I don't know the 430.

-----Original Message-----
X-from: rdpierce-at-pobox.com [mailto:rdpierce-at-pobox.com]
Sent: Sunday, July 21, 2013 4:21 AM
To: wesaia-at-iastate.edu

I'm completely confused by the numbers I'm getting from my S430. My understanding is that beam current measures electrons in the column, and probe current measures electrons striking the sample. Probe current is less than beam current because things like apertures prevent all the electrons in the column from striking the sample.

Now I would think that for a given column configuration, probe current and beam current have to increase or decrease together. I'm not seeing that, and I don't understand it.

I've set up a data zone with both probe current and beam current displayed. When I increase or decrease the probe current, I see the secondary electron image get lighter or darker, as I expect, but beam current stays the same.

Adjusting beam current isn't intuitive. I have to open a status box and add it as a data field, but then I can type it in. It seams the maximum beam current is 400 uA. When I change the value, the image gets darker or lighter as I expect, but the probe current doesn't change!

To make matters more interesting, I can change the filament current, and it makes the image lighter or darker. I imagine as the temperature of the filament changes, its thermionic emission changes, causing more or less electrons to be released, hence a brighter or darker secondary electron image. But wouldn't that change both beam and probe current? I change this and both beam and probe current as reported by the S430 stay the same.

I'm completely confused! These numbers on the S430 don't seem to match what I expect.

Also, my understanding is that one can measure probe current by shooting the beam into a faraday cup and measuring the resulting current as it leaves the stage. While I don't have one of these yet, I do see the ground for the stage coming through a feedthrough in the front door. What kind of connector is it, and where can I find one? It turns out the hackerspace has a Keithley electrometer that can measure picoamps, and I'm guessing I can hook this inline somehow. (That takes a BNC triax connector, which I'd need to find.

Any thoughts?

Regards,
Ryan

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21, 37 -- From wesaia-at-iastate.edu Sun Jul 21 15:15:23 2013
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21, 37 -- From: "Straszheim, Warren E [BIOTC]" {wesaia-at-iastate.edu}
21, 37 -- To: "rdpierce-at-pobox.com" {rdpierce-at-pobox.com}
21, 37 -- CC: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
21, 37 -- Subject: RE: [Microscopy] SEM: Beam / probe current measurement on a Leica
21, 37 -- S430
21, 37 -- Thread-Topic: [Microscopy] SEM: Beam / probe current measurement on a Leica
21, 37 -- S430
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I have a couple of suggestions to consider.

The first suggestion is simply to try the O2-Ar mix. You may be surprised
how well a carbon support film will hold up to an O2 plasma. I was. Try a
two minute run at first and work it up to a five minute run.

The second suggestion is to simply try an Ar plasma. Replace the mixture
gas that you are using with the Fischione unit with just pure Ar. The Ar by
itself may give you what you need.

The third suggestion is to try a mixture of H2, O2 and majority Ar. I would
try about 10% O2, 10% H2, and balance Ar. This should not be dangerous.
When a mixed gas plasma comes on, the more easily ionized gases will ionize
first at lower power. As all of the gas is ionized, then the less ionizable
gaes will kick in. As you raise the power, the easy gases go first followed
by the less easy. I'm guessing here because I don't want to look up the
numbers, but I would guess that O2 would be the easiest, followed by H2,
followed by Ar. Now in a plasma cleaning system, you typically will operate
at low power and what gases get ionized will depend on their concentrations.
You can easily order a lecture bottle of a gas mixture to try this out.

-Scott Walck
Sr Scientist
Bowhead Science and Technology, Inc.
Army Research Laboratory
Aberdeen Proving Ground, MD

-----Original Message-----
X-from: one_twinklestar-at-yahoo.com.sg [mailto:one_twinklestar-at-yahoo.com.sg]
Sent: Sunday, July 21, 2013 12:26 PM
To: S.Walck-at-comcast.net

Dear All, may i know if you
have any advices for cleaning the samples that are supported by the carbon
films on the copper grids? I did find some advices from the past emails in
the mailing list. Any comments on using H2 /O2 gases as what is implemented
in Gatan Solarus? As i am using Fishione Nanoclean, i am thinking if getting
H2 gas with O2 gas will yield the same result. Due to the safety reasons in
lab, the safety officer suggested that i get a
H2 generator rather than H2 gas cylinder, but it does cost quite an
investment. I would like to thank Fishione , which also kindly provide me
some recipe on this cleaning too (but not based on H2/O2). On the other
hand, though, I am hoping if someone could share some advices on this too?

Cheers,
Yee Yan, Tay
FACTS LabNTU, Singapore

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Hello,
Thank you all who sent me suggestions and advices to overcome our
problem..
Last week I did some vacuum tests. The results and the answers to the
questions raised in your emails will be available on the link to web
page, hopefully this week.

However, we are still in trouble with IGP.

With best regards

Oldrich

On Mon, 15 Jul 2013 08:19:22 -0500 benada-at-biomed.cas.cz wrote:

}
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} Hello all,
} Please, could anybody give us an advice or hint on following problem?
} After cathode (W) exchange in our Philips CM100, we are not able to
} get ultra high vacuum. IGP pump switch off and on repeatedly and also
} the valves V3 and V4 are closed and open in the same cycle.
} The LED on IGP HT source is lighting all the time.
} I have taken two snaps of OPCON VACUUM page. The link for images is:
}
} http://www2.biomed.cas.cz/~benada/IGP_trouble2013.html
}
} Thanking in advance for any response.
}
} Best regards Oldrich
}
} --
} Oldrich Benada
} Institute of Microbiology AS CR. v.v.i.
} Videnska 1083
} 142 20 Prague 4
} Czech Republic
}

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7, 39 -- From benada-at-biomed.cas.cz Mon Jul 22 02:26:42 2013
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Hi Ryan

I often use the Leica 430 and the Zeiss range of SEM on my courses, so I may
be able to help you.

Let us consider the parameters that should change the probe current, which
we assume is related to the number of electrons striking the specimen.

1. Filament position - the further back the filament from the cathode
cap the lower the emission level; known as beam current or emission current.
2. Bias Setting - the higher the applied bias field the lower the
emission current but that is in relation to the filament position. For
example with the filament very close to the cap the emission will increase
unless the bias field is used to reduce that emission. Changing the current
in the software simply changes the bias allowing more or less emission
current.
3. The setting of the first condenser lens current, the higher the
current the smaller the number of electron passed on from the first to the
second condenser lens. Remember the probe is reduced in size, and therefore
current, by the second condenser lens only taking the centre of the beam
presented to it, the remainder being trapped on the spray apertures
4. The size of the beam defining aperture.

In truth the probe current is a crude calculation that assumes a great deal.

In relation to 1 above the manufacturer assumes a specific filament
position; probably totally untrue for most people's gun settings.

In relation to 2 above with so many variables this seems to be ignored.

In relation to 3 above this is the variable that is triggering a change in
the probe current readout in the software.

In relation to 4 above if the software asks for information on the size of
this aperture they are using this data along with the first condenser
current to vary the probe current readout.

The manufacturer calculates the probe current when designing the instrument,
assuming that the filament is set in a specific position in the cathode and
the emission current is at a recommended value (setting the gun bias level).
The software contains this information but apart from the change in first
condenser and perhaps beam defining aperture no further software adjustments
take place. You will judge from your experiments that what I relate is
correct?

If you wish to see the true probe current, where all of the variables I have
discussed have an effect, buy a Pico Ammeter and set this up with a Faraday
Cup using the "bnc" connector mounted on the stage door.

I hope this helps because everywhere I go people ask the same question and I
have to say "it's just a crude guess by the manufacturer".

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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realname - Skalli Abdeljabar
Email - skalli01-at-yahoo.ca
SUBJECT_OF_QUESTION - Asbestos microscope
QUESTION - Hi,
I'm looking for a used asbestos microscope in Canada.
Do you know if they are some suppliers in Canada.

Thank you

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Title-Subject: [Filtered] LM Inverted Microscope

Message: Our company is looking to purchase a new inverted microscope for the purpose of examining
nanoparticles. This would likely be done in darkfield at 50X to 100X. Does anyone have suggestions
for what they have and like, or maybe more importantly what they don't like. Thanks in advance.

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Title-Subject: [Filtered] Resolution limits in Electron Microscopes

Message: Hello to all,

I just came across this article published today, based on a PRL paper, and thought this is an
interesting discussion topic. Did anyone see it before?

http://physics.aps.org/articles/v6/82

Marcela.

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Dear Microscopists,

If you are planning to attend M&M 2013, we welcome you to Indianapolis and extend a
special invitation to attend our informative tutorials and presentations covering the following topics:

Poster Presentation: Monday, Aug. 5th, 3:00 pm
A17.P1 Vendor Symposium, Exhibit Hall: Poster Presentation No 57. FS-8500 Provides Superior Specimen Preparation - Dave Bentley and Geon Seo, PhD.

Freeze Substitution Tutorial and Demonstration: Monday, Aug 5th, 5:45 pm - 6:30 pm
At RMC Booth No 538, freeze substitution tutorial and demonstration. Seating is limited, so please sign up as early as possible at the MSA Megabooth on Monday afternoon before the exhibit hall closes.

Live demonstrations of Cryo-Ultrathin Sectioning of biological and material
science samples at LN2 temperatures including cryowet sectioning of
polymers. Visit us at Booth No 538 any time to discuss your application and
schedule a demonstration.

Looking forward to seeing you in Indianapolis.

Kind regards

--
Dave Roberts
Director-RMC Products
Boeckeler Instruments Inc
4650 S Butterfield Drive
Tucson, Arizona 85714
Tel: 520-745-0001
Fax: 520-745-0004
www.rmcproducts.com
Skype:dave.robertsrmc

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Colleagues

Please take notice of the non-advertising rules of the Microscopy
Listserver.

Announcing Posters or Live demo's at upcoming meetings
(regardless of their location) amounts to using the Listserver
for commercial advertising. This information is not appropriate
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If anyone is interested in the scientific program and/or
listings of platform/poster/tutorial presentations please
visit the respective conference/meeting WWW pages. You can
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For the M&M meeting that would be:

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The complete scientific program and listing of all events
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Thanks for your cooperation.

Your Friendly Neighborhood "Syscop"
Nestor

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The box said ...
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So I bought a Mac !

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Colleagues,
A group of avid golfers are organizing an informal outing to a local course early in the morning of Sunday August 4th. We currently have 6 participants. We propose to visit a course where clubs will be available for rent.
If anyone is interested in participating please let me know ASAP as we will soon reserve tee times. Also please let me know if you need or can provide transport to and from the course. We have several volunteers already.

Hit 'em straight!

Chris

Christopher J. Gilpin Ph.D.
Director, Life Science Microscopy Facility
Purdue University
Whistler Hall of Agriculture Research, Room S052
170 S. University St
West Lafayette, IN 47907
765-494-7750

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6, 28 -- From gilpin-at-purdue.edu Tue Jul 23 13:35:25 2013
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I have decided to drive to Indianapolis for the M&M meeting and I am looking
for someone that might like to share the trip with me and do some of the
driving. I live in Havre de Grace, Md and will be leaving on Saturday
morning and will be returning Friday morning the week of the meeting. I am
planning on taking the I-76 & I-70 route, but could take the I-68 & I-70
route. Anybody interested in taking this trip with me? No smokers, please.

-Scott Walck

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3, 33 -- Subject: Need a co-pilot to M&M 2013
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Email: henrik.kaker-at-guest.arnes.si
Name: Henrik Kaker

Organization: SEM-EDS Lab at Metal Ravne

Title-Subject: [Filtered] Question about the vacuum in the old Jeol JSM 35CF (32 years)

Message: Dear All,

I was measured the vacuum in the Jeol SEM and the results are:

Anode (gun) chamber: 2.50 x 10 exp (-4) Torr
Sample chamber: 8.34 x 10 exp (-5) Torr

Is this a normal situation, that the vacuum in the anode chamber is higher then in the sample chamber.
I was also cleaned the SEM column and diffusion pump and replaced the oil in DP.

Best regards,

Henrik

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In Remembrance

Peter Roland Swann
February 4, 1935 - July 14, 2013

Peter was a gentleman in all the senses of the word. He trusted those around him, and helped them
achieve things they did not always know they were capable of. He was a hard worker, talented
inventor and a great organizer, extracting order out of chaos in any field he entered. He also had
a wonderful sense of humor, and great gifts for going straight to the heart of the matter and for
keeping things in perspective. He was always inspiring to work with, and he will be greatly missed.

Peter�s scientific career started in Cambridge UK with a Ph.D. in metallurgy, a field that remained
his first scientific love. He then became a researcher at the US Steel�s Edgar Bain Fundamental
Research Laboratory in Monroeville near Pittsburgh. Among the many discoveries and inventions Peter
made at US Steel was a double tilt sample holder for a Siemens Elmiscope 1 electron microscope.
Siemens, the premier manufacturer of electron microscopes at the time, became interested in Peter�s
design, and in 1964 Peter formed the company Gatan with his brother Rex to supply the holders to
Siemens and to develop other devices for electron microscopy.

For the next one and half decades, Peter managed to combine a research career that culminated in a
full professorship at Imperial College, London, with the role of the chief designer at Gatan. This
led to long working hours and a large number of outstanding scientific contributions. In 1978 he
came to Pittsburgh to direct Gatan full time. Gatan then really took off, especially after 1983,
when Peter relocated its headquarters to the San Francisco Bay Area, and after �going international�
with German and UK offices.

Electron microscopy has aptly been called �the eyes of science�. Thanks to Peter�s work, the eyes
have become more open, more sensitive, and more sharply focused. As one example, Peter designed
many different types of sample holders that allow scientists to look at samples that are cooled,
heated, strained, in a gaseous environment rather than in vacuum � i.e., under the conditions they
encounter in the real world, rather than sitting rigidly as if they were in a photographer�s studio.
This is now called in-situ microscopy, and it has greatly contributed to our scientific knowledge
of the �real world�. As another example, Peter pioneered using ion beams to micro-machine samples
such as semiconductor chips, a development adopted by others with a huge impact on modern
electronics and other fields. Gatan also became the leading supplier of specimen preparation
equipment, pioneered electronic recording of microscopy data that greatly improved on photographic
film, and developed a line of widely used electron energy loss spectrometers and imaging filters.
It also introduced image processing software that has now become the standard software that is
supplied with all the leading electron microscopes, no matter whether they are made in Japan,
Holland, Germany, the Czech Republic or the USA. In many ways, under Peter�s guidance, Gatan became
the Cisco, Apple and Google of electron microscopy, all in one.

Peter�s guiding principle was KISS � keep it small and simple. Another one was an insatiable
curiosity about how things worked and how they could be made better. He would spend hours refining
each new invention to purge superfluous elements such as screws that were not really needed, and in
the process he would achieve a Zen-like purity of design. His interests ranged over many different
areas of science and human endeavor in general. In each area, he quickly zeroed in on the essential
and was bubbling with ideas about how to improve it.

Peter touched many things and made them better, and he was not shy about sharing his ideas. We used
to joke that when he arrives at the Pearly Gates, he will say to the Peter with the halo above his
head: �Very nice design, Saint Peter, but have you thought about making the path straighter by
moving the Gates a little to the left, simplifying the supports to make them stronger, and �?�
Those of us lucky enough to see the Pearly Gates ourselves one of these days will be struck by their
perfection, and we will suspect that Peter may have had a hand in it.

Ondrej Krivanek

--------------------------------
---===[|]===---
--------------------------------

I will echo Ondrej's thought's above.
Well said.

Nestor
Your Saddened Neighborhood SysOp

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Email: hzhan-at-live.com
Name: Hong ZHAN

Organization: Institute de Biologie de l'Ecole Normale Superieure

Title-Subject: [Filtered] Question about chemical fixation of C. elegans for TEM

Message: Dear all,

I have a very stupid question about chemical fixation protocol of C. elegans for TEM.

I am not using chemical fixation protocol very often for C. elegans but High-pressure freezing. Here
is my question: whether I could immobilize worms inside an agarose pad then add fixative? Agarose
would not cross-react with fixative? Some published protocols use agarose to help orientate worms
after treatement with 2% Osmium tetroxide I would like to first put worms in agarose, then remove
heads and tails and add fixative.

For fixative I would choose glutaraldehyde and osmium tetroxide. Cause our lab used to use it long
time ago.

Thanks a lot.

Best,

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Hello Listers,
We've been trying to process some liquid crystals for TEM and I am
running out of ideas. I hope someone can help.

So far we have tried the vitrobot and cryo-stage TEM. This works fine
but hasn't yielded any useful images.
I've also tried to do some cryo sections of a thin layer of the
crystals sandwiched between aclar films. I'm not satisfied that we've
found the right sectioning conditions but each attempt has had
problems getting even sections and often separating the sandwich.

I'm wondering if there is a way to chemically fix the samples, similar
to a biological method but my searches haven't turned anything up.

I am open to any and all suggestions so please suggest away!

Jay Campbell

Materials Science Center
UW Madison
1509 University Av
Madison WI 53706

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Name: Glenn Poirier

Organization: Canadian Museum of Nature

Title-Subject: [Filtered] Removal of gold coating from analytical samples

Message: Dear Colleagues,
Some researchers at my university are searching for a way to remove a gold coating from analytical
samples.
The samples consist of Al discs containing indium plugs into which individual zircon grains have
been pressed.
I don't want to use iodine in KI because it reacts with the Al. Mechanical polishing is probably out
because of the indium (don't ask about my attempts to polish indium!).
Does anyone know of any other possibilities I should look into. I seem to remember this discussed
before, but I can't find anything in the archive. Any help gratefully appreciated.

I subscribe to the list but my employer insists on attaching advertising to the bottom of my email
so my emails to the server get rejected. I'll see any answers posted to the list

Cheers

Glenn Poirier
University of Ottawa/Canadian Museum of Nature Microanalysis Facility
Ottawa

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Good morning,
dear Glenn,
don't know whether this will help anyway....
since you said:
} I seem to remember this discussed before, but I can't find anything in
} the archive. Any help gratefully appreciated. {:

Searching my files the following MSA-Listserver-Threads appeared to contained the words

"polishing", or "Indium"

[Microscopy] TEM - room temperature cross-section sample prep [cross-section samples of inorganic solar cells] (Q: 2013-05-30, no answer found)

[Microscopy] Etching Stainless Steel 316 (Q: 2012-12-18)

[Microscopy] Gallistan in SEM
{Materials: Gallistan [= [Galinstan] (Gallium, Tin, Indium) ] in SEM [:is there an Outgassing problem?]}
(Q: 2011-12-07)

[Microscopy] EDX on Sulfur [ratio differences with SDD vs SiLi and kV] (Q: 2011-03-04)

[Microscopy] small system for ITO (Indium Tin Oxide) coatings (Q: 2005-09-27 )

If you could specify a little bit more a detail of the discussion you can remember, this would help....

Best regards,
greetings from Salzburg

Wolfgang

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} Email: gpoirier-at-mus-nature.ca Name: Glenn Poirier
} Organization: Canadian Museum of Nature
} Title-Subject: Removal of gold coating from
} analytical samples
} Message:
}
} Dear Colleagues,
} Some researchers at my university are searching for a way to remove a
} gold coating from analytical
} samples. The samples consist of Al discs containing indium plugs into
} which individual zircon grains have
} been pressed.
} I don't want to use iodine in KI because it reacts with the Al.
} Mechanical polishing is probably out because of the indium (don't ask
} about my attempts to polish indium!).
} Does anyone know of any other possibilities I should look into?
} I seem to remember this discussed before, but I can't find anything in
} the archive. Any help gratefully appreciated.
}
} I subscribe to the list but my employer insists on attaching
} advertising to the bottom of my email so my emails to the server get
} rejected.
} I'll see any answers posted to the list
}
} Cheers
}
} Glenn Poirier
} University of Ottawa/Canadian Museum of Nature Microanalysis Facility
} Ottawa
}
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I use to use a small droplet of KCN. I put a drop of distilled water on the surface and scoot in a few crystals of KCN. The sodium salt works too, but takes longer.

I had a fine glass rod with a even smaller glass ball on the end. I used that for microchemical testing and I would touch the rod to the top surface of the droplet and "scrub" the surface via surface tension.

I never worried about the toxic effects. The total concentration was so small and good lab practices keep you safe, but you should check the MSDS. Disposal? We did so little of this we just flushed it down the drain.

Stay safe...........

Frank

-----Original Message-----
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Sent: Thursday, July 25, 2013 6:49 PM
To: Frank Karl

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Email: gpoirier-at-mus-nature.ca
Name: Glenn Poirier

Organization: Canadian Museum of Nature

Title-Subject: [Filtered] Removal of gold coating from analytical samples

Message: Dear Colleagues,
Some researchers at my university are searching for a way to remove a gold coating from analytical
samples.
The samples consist of Al discs containing indium plugs into which individual zircon grains have
been pressed.
I don't want to use iodine in KI because it reacts with the Al. Mechanical polishing is probably out
because of the indium (don't ask about my attempts to polish indium!).
Does anyone know of any other possibilities I should look into. I seem to remember this discussed
before, but I can't find anything in the archive. Any help gratefully appreciated.

I subscribe to the list but my employer insists on attaching advertising to the bottom of my email
so my emails to the server get rejected. I'll see any answers posted to the list

Cheers

Glenn Poirier
University of Ottawa/Canadian Museum of Nature Microanalysis Facility
Ottawa

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28, 29 -- From frank_karl-at-ardl.com Fri Jul 26 06:59:30 2013
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If polishing or KCN etching are not compatible with the sample material,
and if sample is either sufficiently small or small fraction can be
broken off from it to fit into the ion mill, then I'd try broad-beam Ar
or Xe milling....

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
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On 7/26/2013 8:00 AM, frank_karl-at-ardl.com wrote:
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} I use to use a small droplet of KCN. I put a drop of distilled water on the surface and scoot in a few crystals of KCN. The sodium salt works too, but takes longer.
}
} I had a fine glass rod with a even smaller glass ball on the end. I used that for microchemical testing and I would touch the rod to the top surface of the droplet and "scrub" the surface via surface tension.
}
} I never worried about the toxic effects. The total concentration was so small and good lab practices keep you safe, but you should check the MSDS. Disposal? We did so little of this we just flushed it down the drain.
}
} Stay safe...........
}
}
} Frank
}
} -----Original Message-----
} X-from: microscopylistserver-noreply-at-microscopy.com [mailto:microscopylistserver-noreply-at-microscopy.com]
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} To: Frank Karl
} Subject: [Microscopy] viaWWW:Removal of gold coating from analytical samples
}
}
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}
} Message: Dear Colleagues,
} Some researchers at my university are searching for a way to remove a gold coating from analytical
} samples.
} The samples consist of Al discs containing indium plugs into which individual zircon grains have
} been pressed.
} I don't want to use iodine in KI because it reacts with the Al. Mechanical polishing is probably out
} because of the indium (don't ask about my attempts to polish indium!).
} Does anyone know of any other possibilities I should look into. I seem to remember this discussed
} before, but I can't find anything in the archive. Any help gratefully appreciated.
}
} I subscribe to the list but my employer insists on attaching advertising to the bottom of my email
} so my emails to the server get rejected. I'll see any answers posted to the list
}
} Cheers
}
} Glenn Poirier
} University of Ottawa/Canadian Museum of Nature Microanalysis Facility
} Ottawa
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Contents Retrieved from Microscopy Listserver Archives
http://www.microscopylistserver.com/MicroscopyListserver/MicroscopyArchives.html

Dear All, thank you very much for your kind sharing of the information you have! I will go and try it!!

Cheers,
Yee Yan

----- Original Message -----
X-from: "S.Walck-at-comcast.net" {S.Walck-at-comcast.net}
To: one_twinklestar-at-yahoo.com.sg
Cc:
Sent: Monday, 22 July 2013, 7:21

I have a couple of suggestions to consider.

The first suggestion is simply to try the O2-Ar mix.� You may be surprised
how well a carbon support film will hold up to an O2 plasma.� I was.� Try a
two minute run at first and work it up to a five minute run.

The second suggestion� is to simply try an Ar plasma.� Replace the mixture
gas that you are using with the Fischione unit with just pure Ar.� The Ar by
itself may give you what you need.

The third suggestion is to try a mixture of H2, O2 and majority Ar.� I would
try about 10% O2, 10% H2, and balance Ar.� This should not be dangerous.
When a mixed gas plasma comes on, the more easily ionized gases will ionize
first at lower power.� As all of the gas is ionized, then the less ionizable
gaes will kick in.� As you raise the power, the easy gases go first followed
by the less easy.� I'm guessing here because I don't want to look up the
numbers, but I would guess that O2 would be the easiest, followed by H2,
followed by Ar.� Now in a plasma cleaning system, you typically will operate
at low power and what gases get ionized will depend on their concentrations.
You can easily order a lecture bottle of a gas mixture to try this out.�

-Scott Walck
Sr Scientist
Bowhead Science and Technology, Inc.
Army Research Laboratory
Aberdeen Proving Ground, MD

-----Original Message-----
X-from: one_twinklestar-at-yahoo.com.sg [mailto:one_twinklestar-at-yahoo.com.sg]
Sent: Sunday, July 21, 2013 12:26 PM
To: S.Walck-at-comcast.net

Dear All, may i know if you
have any advices for cleaning the samples that are supported by the carbon
films on the copper grids? I did find some advices from the past emails in
the mailing list. Any comments on using H2 /O2 gases as what is implemented
in Gatan Solarus? As i am using Fishione Nanoclean, i am thinking if getting
H2 gas with O2 gas will yield the same result. Due to the safety reasons in
lab, the safety officer suggested that i get a
H2 generator rather than H2 gas cylinder, but it does cost quite an
investment. I would like to thank Fishione , which also kindly provide me
some recipe on this cleaning too (but not based on H2/O2). On the other
hand, though, I am hoping if someone could share some advices on this too?

Cheers,
Yee Yan, Tay
FACTS LabNTU, Singapore

==============================Original Headers==============================
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Email: hzhan-at-live.com
Name: Hong ZHAN

Organization: Institute de Biologie de l'Ecole Normale Superieure

Title-Subject: [Filtered] Re:Question about chemical fixation of C. elegans for TEM‏

Message: Thanks Dr. Hall.
It is true that removing worms'heads and tails are difficult when they are awake. I tried using
levamisol to put them down first. I did in vivo probe labeling for target protein in worms, so that
I want to try like this way. I am waiting for HPF machine cause it has some problem. But I have
tried once, put worms with drugs and embedded in agarose do CLEM protocol. I sealed freezing chamber
with Hexadacene, it was nice (according to Kolotuev I et al. publication ). However, I have some issues:
worm after HPF (Leica HPM 100) and fixation, embedded in araldite . but worm was cracked at some
parts.... I only have dorsal or ventral sides... I would like to know whether you have met same
problem and whether you have some tips for avoiding that?

Many thanks for your reply
Have nice weekend

Best,

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Hello,

Partly in reply to Hong Zhan's question: I don't think cracks in HPF frozen material have been reported in specimens observed at a temperature below the devitrification or recrystallisation temperature of water. Neither have cracks been described in freeze fracture replicas of high pressure frozen specimens. It is therefore likely that those cracks are the result of follow-up procedures.

After HPF the available water is partly (at least) frozen to either high density ice or is in a high density amorphous state. These high density conditions are lost during freeze substitution. It is thought that high density forms have changed to low density already at the onset of freeze substitution and this would cause a change in volume at ambient pressure (from ~1.16 to ~0.92 g/cm3), the tension giving rise to cracks. This has been suggested a.o. by Prof Moor and Daniel Studer in their publications and presentations.

Allow me to add a speculation: the problems may become more serious as a result of the embedding method, especially when higher temperatures were used for polymerisation. Proteins will change conformation as a result of elevated temperatures, especially if still hydrated. I am thinking of nicely curled up fried bacon! On a micro-scale this might very well lead to further cracking of pre-existing rips.

But you asked if one can do something about this.
Freeze substitution: physics can't be changed, and higher temperatures for freeze substitution are thought not to give sufficient removal of ice in a practical time span. I do not know if anyone tried to do this, by the way. The rate for removing water molecules from ice in a vacuum is not necessarily the same as in a medium. The solvent molecules will interact with water molecules as well and may bind water, possibly allowing for better removal rates than anticipated. Results obtained with Kent McDonald and Rick Webb's procedures support this. Having said that, one is bound to the freezing point of the medium of course.
Embedding: people who have worked with lower temperature embedding methods may have had better results, with less pronounced cracks. I personally have no experience, but it seems a sensible way to go to minimise cracks.

Having said that, time for breakfast in New Zealand! Crumpled protein ....

Jan Leunissen

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Hello

I often see a weird ring in the DP when I do selected-area diffraction.
We have a JEOL 2010 LaB6 electron microscope. I talked this to JEOL
engineers, but
they do not know either.
Does anybody have ever met this problem? Or anyone knows what's wrong?

Thank you very much

--
Dr. Yueling Qin
Senior Research Support Specialist
UB2020 Integrated Nanostructured Systems Initiative,
University at Buffalo, SUNY, Buffalo, NY 14260,
Phone (716) 645-8698

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You are going to have to describe it better or post a link to an image of
it. Prepare to describe your sample. An image of the area that you have
selected would also work. Is it only happening with the one sample?
Remember, the shape of the "spot" in reciprocal space will be "reciprocally"
related to the shape of any phases that are diffracting. If you have a
short dimension in the image of a phase, then it will be a large dimension
in reciprocal space and the Ewald sphere intersecting this shape can look
"weird".

-----Original Message-----
X-from: yqin-at-buffalo.edu [mailto:yqin-at-buffalo.edu]
Sent: Sunday, July 28, 2013 6:48 PM
To: S.Walck-at-comcast.net

Hello

I often see a weird ring in the DP when I do selected-area diffraction.
We have a JEOL 2010 LaB6 electron microscope. I talked this to JEOL
engineers, but they do not know either.
Does anybody have ever met this problem? Or anyone knows what's wrong?

Thank you very much

--
Dr. Yueling Qin
Senior Research Support Specialist
UB2020 Integrated Nanostructured Systems Initiative, University at Buffalo,
SUNY, Buffalo, NY 14260, Phone (716) 645-8698

==============================Original Headers==============================
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We get that too, on our JEM 2100 LaB6. In our case, I'm pretty sure it comes from the selected-area diffraction aperture. The ring shows up in a slightly different plane from the objective lens BFP, so it can usually be suppressed by changing the intermediate lens (diffraction) focus a little, but sometimes not completely, especially if the signal you are looking for is weak. I think it is a problem with the apertures JEOL uses. Not sure if these are Mo or Pt. I can't find an example at the moment, and we are waiting for the service engineer to come today to fix the instrument, but I can send you something for comparison in a day or two if it is working again.
------------------------------------------
Phil Ahrenkiel, Associate Professor
Nanoscience and Nanoengineering Ph.D. Program
South Dakota School of Mines and Technology
501 E. Saint Joseph St.
Rapid City, SD 57701 U.S.A.
Office: EP 221
Phone: 605-394-5238
Email: Phil.Ahrenkiel-at-sdsmt.edu

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Hello

I often see a weird ring in the DP when I do selected-area diffraction.
We have a JEOL 2010 LaB6 electron microscope. I talked this to JEOL engineers, but they do not know either.
Does anybody have ever met this problem? Or anyone knows what's wrong?

Thank you very much

--
Dr. Yueling Qin
Senior Research Support Specialist
UB2020 Integrated Nanostructured Systems Initiative, University at Buffalo, SUNY, Buffalo, NY 14260, Phone (716) 645-8698

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HI all,

Thank you for your attention. I attach a link below to a picture, which
was obtained by SAD over an empty area without specimen.

I will follow the suggestions to exam the effects of C2 and
selected-area apertures.

https://lh3.googleusercontent.com/-7yzBRtoBFRA/Ufa2YuOhjiI/AAAAAAAAADQ/aTdZmOMiXHc/s821-no/DIFF+RING.jpg

--
Dr. Yueling Qin
Senior Research Support Specialist
UB2020 Integrated Nanostructured Systems Initiative,
University at Buffalo, SUNY, Buffalo, NY 14260,
Phone (716) 645-8698

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Dear listers,

I have a researcher that is doing their own EM. He recently used a protocol where the fixative has safranin O added. He now has a problem with the sections washing off the grids when he is staining and rinsing them, which was not a problem before. Has anyone out there used safranin O in their EM protocols and encountered this problem? I'm guessing he is going to have to put some kind of adhesive on the grids, any recommendations about grid adhesive would be appreciated also.

Tom Bargar
UNMC
Electron Microscopy Core Facility
986395 Nebraska Medical Center
Omaha, NE 68198-6395
402-559-7347
tbargar-at-unmc.edu

The information in this e-mail may be privileged and confidential, intended only for the use of the addressee(s) above. Any unauthorized use or disclosure of this information is prohibited. If you have received this e-mail by mistake, please delete it and immediately contact the sender.

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Email: plarson-at-ou.edu
Name: Preston Larson

Organization: University of Oklahoma

Title-Subject: [Filtered] A compact heating substage for temperatures up to 1200�C

Message: Hi all,

We are looking for sample heaters compatible with FE-SEMs.

Something along the lines that Fullam used to make would be great.

For example, the Fullam P/N 18020 -A compact heating substage for temperatures up to 1200�C- would
be appropriate.

Unfortunately, Fullam no longer makes these heaters and MTI, which has taken over much of Fullam's
product line here in the US, seems to have discontinued this model.

Does anyone have such a heater sitting around and want to sell it to us?

Does anyone know of a comparable heater stage still being made?

Feel free to contact me off list.
Thanks in advance,
preston

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Dr. Qin

When I worked at HP we had a very lovely JEOL 2500 TEM. Although the 2500 is a Field Emission Gun (FEG), we used to see something similar. Mr. Tamba, JEOL service based in LA fixed it for us. Rather, Tamba gave us the knowledge to fix it ourselves! We fixed it by always performing selected area diffraction (SAD) left of crossover, where the beam is more parallel. You know what they say about free advice ;0), but I believe that by operating left of crossover instead of right of, you shall get rid of your mystery ring! You could also have your East coast JEOL give Tamba-san a call to confirm this for your LaB6 JEOL. Short of that it is very easy to try, just give that intensity knob a little twist to the left ;0)

Pete Eschbach Ph.D.
Director of The Electron Microscope Facility
Oregon State University
Peter.Eschbach-at-oregonstate.edu
541 737 5645

Sent from my iPad

On Jul 28, 2013, at 3:48 PM, yqin-at-buffalo.edu wrote:

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} I often see a weird ring in the DP when I do selected-area diffraction.
} We have a JEOL 2010 LaB6 electron microscope. I talked this to JEOL
} engineers, but
} they do not know either.
} Does anybody have ever met this problem? Or anyone knows what's wrong?
}
} Thank you very much
}
} --
} Dr. Yueling Qin
} Senior Research Support Specialist
} UB2020 Integrated Nanostructured Systems Initiative,
} University at Buffalo, SUNY, Buffalo, NY 14260,
} Phone (716) 645-8698
}
} ==============================Original Headers==============================
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} 4, 32 -- Subject: Weird Diffraction Ring
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All,

I'd like to acquire a SiLi EDS detector that will fit a JEOL 2010 TEM. Any make. Working or not. Donations preferred. I'll happily pay shipping. Please contact me offline.

Thanks, Owen

Owen Mills
Michigan Tech University

==============================Original Headers==============================
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X-from: jefferson.bettini-at-lnnano.cnepm.br ()

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Email: jefferson.bettini-at-lnnano.cnepm.br
Name: Jeffeson Bettini

Organization: LME/LNNano/CNPEM

Title-Subject: [Filtered] job opportunity

Message: LNNano hires Researcher to work at the Electron Microscopy Laboratory.

The Brazilian National Nanotechnology Laboratory as part of the National Center for Research in
Energy and Materials (www.lnnano.cnpem.br) is with two open fixed position for researcher in their
Electron Microscopy Laboratory (EML). The ELM operates as open National Facility for multiuser in
Brazil.

For this position it is essential to have experience in Transmission Electron Microscopy.

The candidate is invited to develop research in a specific field of electron microscopy, such as
holography, tomography, spectroscopy, advanced electron diffraction Quanti(S)TEM, etc. The research
can be developed collaboratively with our users or independently. Furthermore, the candidate is
invited to collaborate to maintain and continuously improve this facility, which is equipped with
three Transmission Electron Microscopes, a Dual Beam microscope and three Scanning Electron microscopes.

Requirements:

Training in Physics, Chemistry or Engineering with full doctorate in the area.

English: fluent.

Interested send resume to gabriela.carvalho-at-cnpem.br and in the subject put the job title: LNNano
Researcher LME-LNNano. Note: Resumes without identifying in the subject field will be automatically
disregarded in the selection process.

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We have a FEI Tecnai 12T with a Gatan side-entry digital camera.

Description: Model 780 DualView 1300 x 1030 12-bit Air-cooled Peltier 35-mm port
Damage: "Snap, crackle, and pop" during a session with the normal acrid odor of burning electronics.

We have Gatan software and the apparently unharmed Roper control box.

We cannot get our camera repaired - even the manufacturer will not touch it at this point in time - so we are looking for one that works, but has been retired with no future purpose. Given the difference between new and repair, we would likely entertain any reasonable offer. {As well as an offer for any reasonable replacement}.

We have been told that the vacuum in the device was expected to be maintained by periodic checks.

Even a rented loaner while we try to resolve the normal institutional issues with expensive replacements, service, etc. would be most welcome.

We cannot resort to film as we no longer have a usable darkroom.

Our primary use is high resolution and eDiffraction.

Thanks for your consideration,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

HomePage: http://cmirt.wcupa.edu (with link to instrument Scheduler)

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Email: jacques.werckmann-at-ipcms.u-strasbg.fr
Name: Jacques Werckmann

Organization: IMETRO Brazil

Title-Subject: [Filtered] Quantification by EDSX

Message: Quantification of composition nanopaticules Au / Ag approximately 5nm.
With Noran Thermo Fisher Scientific
The particles are deposited on a film of carbon, electron irradiation causes the formation of a
carbon layer on which the nanoparticles investigated. Although the signal of Au and Ag is detected
after selecting items, no value appears in the table giving the intensities and compositions.
Any help gratefully appreciated

Jacques Werckmann

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Email: BedardJoe-at-Verizon.net
Name: Joe Bedard

Organization: Analytical Answers, Inc.

Title-Subject: [Filtered] TOPCON DS-701 SEM scan module

Message: Hello,

We are currently attempting to find and purchase a working scan PCB
module(Akashi N404MC04), ideally with the proper external output for
separate EDS imaging, for a TOPCON DS-701 SEM instrument. I know this
is a long shot, but
nothing ventured - nothing gained. Please e-mail me with the details
should you have one for sale.

Thank You,

Joe

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Email: info-at-nesmicroscopy.org
Name: New England Society for Microscopy

Organization: New England Society for Microscopy

Title-Subject: [Filtered] Save the Date: NESMs Fall Meeting -at- UMass Lowell

Message: Hello Microscopists,

This is a friendly reminder to save the date for the New England Society
for Microscopys annual Fall Meeting on Thursday, October 17th. The
meeting will be hosted by the University of Massachusetts at Lowell and
will feature facility tours, a buffet dinner, and two technical talks.
Dr. Carol Barry of UML will deliver a lecture on
micro-and-nanostructured surfaces. A second speaker will be announced in
the coming weeks, along with more meeting information. Keep up with NESM
by visiting our website (www.nesmicroscopy.org) and by following us on
Facebook (http://www.facebook.com/NESMicroscopy) and on Twitter
(http://twitter.com/#!/NESMicroscopy).

Cheers,
Rylie Walsh
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Email: dover-at-rpi.edu
Name: Raymond Dove

Organization: Rensselaer Polytechnic Institute

Title-Subject: [Filtered] Gatan 780 CCD Camera Head

Message: I'm looking for a working Gatan Model 780 CCD Dual View Camera
head . I have a working controller.
Short of this......does anyone know of an organization / person who
could repair same.....I'm told this is not possible. Perhaps a
compatible head from another mfg that you know of ??

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abeGood day todayhttp://tanglangchangquan.com/www.cnnnews.com.indexnews41.top.php

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Dear all,

To celebrate the opening of our new TEM facility we organize a 1-day meeting/seminar on a

Inauguration of new NORTEM JEOL microscopes in Trondheim
September 10, 2013
Trondheim, Norway

For more information see:
http://www.ntnu.edu/physics/nortem

Regards,

Ton van Helvoort
IFY, NTNU, Trondheim, Norway

==============================Original Headers==============================
6, 31 -- From a.helvoort-at-ntnu.no Sat Aug 3 06:59:05 2013
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6, 31 -- From: "Antonius T. J. van Helvoort" {a.helvoort-at-ntnu.no}
6, 31 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
6, 31 -- Subject: 1-day seminar advanced TEM, 10 September 2013, Trondheim, Norway
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There is a post-doctoral position in atom probe topography available immediately at Oak Ridge National Laboratory.

For information, please see jobs.ornl.gov, posting "Postdoctoral Research Associate / NB50344514," or shortcut link http://1.usa.gov/14iJjoZ

For questions, primary contact is contact Dr. M. K. Miller (millermk-at-ornl.gov) and secondary contact is Dr. Chad Parish (parishcm-at-ornl.gov).

You may also speak to us at the Microscopy and Microanalysis conference in Indianapolis next week.

Please forward to any qualified applicants.

Thanks,
Chad Parish

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

==============================Original Headers==============================
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10, 31 -- Subject: Post-doctoral position in Atom Probe Tomography
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X-from: Haim H. Bau {bau-at-seas.upenn.edu}

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Email: Haim H. Bau {bau-at-seas.upenn.edu}
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Organization: U Penn

Title-Subject: [Filtered] Position Available

Message: Postdoctoral Position in the Micro/Nano Fluidics Lab at UPenn
The micro/nano fluidics lab at the University of Pennsylvania has an
immediate opening for a postdoctoral fellow to develop devices and
methods for in-situ electron microscopy of objects (i.e.,
macromolecules) suspended in and processes taking place in liquid media.
See
http://bau.seas.upenn.edu/research/in-situ-electron-microscopy-of-liquids-the-nanoaquarium/
The fellow must have a PhD in Engineering or Physics, clean room
(microfab) experience, and exposure to biology.

Outstanding candidates are invited to submit a letter of application
detailing their relevant background, providing the names and e-mail
addresses of at least three references, a CV, and copies of relevant
reprints and pre-prints. Send the application to Professor Haim H. Bau
at meampd-at-seas.upenn.edu.

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With reference to the M&M meeting which starts tomorrow in Indianapolis, I
want to report an issue and a workaround.
The Abstract disk uses a web browser interface. When start.html is opened
in Internet Explorer 8, the body of the table of contents is displayed
against a background image, R:\bin\images\background.png.
This image has a busy scene that has a lot of red. The text in the table of
contents is blue. This is essentially unreadable.
Workaround: use a different browser. I found that the current version of
Google Chrome works OK. The text is in now displayed against a nearly-white
background and is easy to read.
I hope this helps.

To everyone who is coming to Indianapolis, have a safe trip and enjoy the
meeting.

regards,
Don
=============================================
Don Chernoff, Ph.D., President
Advanced Surface Microscopy, Inc.; 3250 N. Post Rd., Suite 120;
Indianapolis IN 46226 USA
E-Mail: donc-at-asmicro.com www.asmicro.com
Voice: 317-895-5630 Toll free: 800-374-8557 (in USA & Canada)
Fax: 317-895-5652
[business activities since 1990: analytical services in AFM, AFM probes,
consulting, training, calibration and test specimens,
calibration and measurement software, used NanoScope equipment.]
=============================================

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PostDoctoral and/or PhD position in software development for electron microscopy at the University of Basel, Switzerland

A postdoctoral and/or PhD position is available at the Center for Cellular Imaging and NanoAnalytics (C-CINA) of the Biozentrum of the University of Basel, Switzerland. Our group is developing the 2dx software package for user-friendly image processing of 2D crystal images. We have recently extended 2dx by a single particle module that treats 2D crystal images as assemblies of correlated individual particles to refine the 3D reconstruction. The goal of this postdoctoral position is to advance these methods for processing of movie-mode recorded cryo-EM image series of semi-2D crystalline and vesicular membrane protein samples, using Bayesian methods, and to interface this package with movie-mode data recording on the Titan Krios / K2 Summit.

The 2dx software package is available at http://2dx.org, and our lab is described at http://c-cina.org. We are fortunate to operate outstanding equipment and our lab studies several highly ordered 2D crystals of membrane protein systems.
The position is funded for several years, and is available immediately. The town of Basel at the Swiss border towards Germany and France enjoys a rich cultural and international atmosphere and beautiful surrounding nature. We are equal opportunity employers. The lab language is English.
For further information, please contact Henning.Stahlberg-at-unibas.ch.

Henning Stahlberg, PhD
Prof. for Structural Biology, C-CINA, Biozentrum, University Basel
Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland
http://c-cina.org | Tel. +41-61-387 32 62 | mailto:Henning.Stahlberg-at-unibas.ch

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9, 30 -- Subject: PostDoctoral and/or PhD position in software development for
9, 30 -- electron microscopy at the University of Basel, Switzerland
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9, 30 -- electron microscopy at the University of Basel, Switzerland
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Listers,

Any labs in/near/around Amarillo, TX with SEM/EDS capabilities? I have
a customer whose EDS went down and needs some samples analyzed. Thanks.

Gary Easton
Scanners Corporation
410-857-7633 x102

-----
No virus found in this message.
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Atomic force microscope and ThermoChemical NanoLithography (TCNL) were used to create a 30 micron wide image of the Mona Lisa by scientists at the Georgia Institute of Technology.
�
Full Story: http://www.dailymail.co.uk/sciencetech/article-2385283/The-Micro-Lisa-Incredible-molecular-painting-masterpiece-times-narrower-HUMAN-HAIR.html

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc /

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X-from: kevinksl-at-att.net ()

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Title-Subject: [Filtered] TEM

Message: Looking an older JEOL TEM; 100C, CX, CXII, 200C, CX, or 1200EX.

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Title-Subject: [Filtered] .lif to .lsm confocal file conversion

Message: Hi all,
I have a colleague who is having the following problem: when we
import Leica .lif files into BitPlane Imaris 7.5, it corrupts the file
when it sends it to AutoQuant X3 for deconvolution, and causes one or
both problems to crash. A friend of mine sent me an .lsm file from the
Zeiss confocal and it worked flawlessly.
We are trying to figure out if it is possible to convert files from
a .lif format into the .lsm file format so they can use the program. He
has had the Imaris program for a while so they will not offer support
without charging a couple of thousand for a technical consulting charge.
Does anyone have any suggestions?
Thanks!
Page

D. Page Baluch, Ph.D.
W.M. Keck Bioimaging Laboratory
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Title-Subject: [Filtered] Z6040 embedding primer ems

Message: Hello All,

Will someone please share their experience or protocol for using the ems
Z6040 embedding primer? I have never used this product before but it
was recommended as a measure to prevent delamination from occurring when
microtoming cadmium sulfide nanowires grown on copper substrate.

Any comments on the above would be greatly appreciated.

marissa

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Hey Everyone,

I encountered a problem while trying to understand
how to analyze low loss STEM EELS spectras.
In the EELS book by Egerton I found the Fourier Log Method
to extract the SSD spectrum.

Now the question:
If understand it correctly, I need the full spectrum (including
the zero loss peak) for the deconvolution.
But is it possible to use a spectra without the Zero loss peak?
Because with the ZL peak I would need to reduce the beam current
and taking EDX spectra simultaneously would become difficult if
not impossible.

Or did I missed something?

Best regards and thanks for the help in advance! :-)

Max

==============================Original Headers==============================
7, 21 -- From raven2020-at-gmx.de Thu Aug 8 08:44:00 2013
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Hi Max,

That's right. One solution I've used to solve this is to acquire the stack twice. On the first shot, I get the zero and low loss region. This scan is usually pretty quick. A second scan then gets the core edge. If you are acquiring simultaneous EDS then acquire it during the core-edge stack since it will be a longer scan.

The problem with this is that you then have to align the two stacks, and it can produce some headaches for figuring out which pixel has the zero loss to match a particular core loss pixel, but it works. For alignment, I have used the ImageJ tools.

Zack Gainsforth
Space Sciences Laboratory, UC Berkeley
7 Gauss Way
Berkeley, CA 94720
work: 510-642-9733
cell: 626-437-9186
zackg-at-ssl.berkeley.edu

On Aug 8, 2013, at 6:53 AM, raven2020-at-gmx.de wrote:

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Hey Everyone,

I encountered a problem while trying to understand
how to analyze low loss STEM EELS spectras.
In the EELS book by Egerton I found the Fourier Log Method
to extract the SSD spectrum.

Now the question:
If understand it correctly, I need the full spectrum (including
the zero loss peak) for the deconvolution.
But is it possible to use a spectra without the Zero loss peak?
Because with the ZL peak I would need to reduce the beam current
and taking EDX spectra simultaneously would become difficult if
not impossible.

Or did I missed something?

Best regards and thanks for the help in advance! :-)

Max

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7, 21 -- From: Max Klingsporn {raven2020-at-gmx.de}
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Pawley's "Handbook of Biological Confocal Microscopy" has already been
suggested to the OP.

***************************************************************************************
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realname - Nicholas Galvan
Email - nicholas.galvan-at-colorado.edu
ORGANIZATION - Colorado State University
EDUCATION - Undergraduate College
LOCATION - Fort Collins, Colorado, USA
SUBJECT_OF_QUESTION - High Speed Microscopy
QUESTION - I am looking for a book that covers the basics of Digital
Microscopy, and a book that would be a good follow up to Jerome Mertz'
book Optical Microscopy. I am hoping that book will cover fraunhofer
diffraction, fresnel diffraction, radiometry, Fluorescence, and confocal
microscopy. Thank you for your time in advanced.

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Email: rodrews-at-gmail.com
Name: Andrew Roholt

Organization: Renovo Neural Inc

Title-Subject: [Filtered] Job posting

Message: Research Associate - 3D-EM Microscopist
This is a full-time position based in Renovo Neural Inc.s (RNI) laboratory in Cleveland, OH. RNI
specializes in preclinical evaluation of drugs for Multiple Sclerosis (MS) and other neural
diseases. This position will support and extend advanced (3D) biological electron microscopy (EM)
services. This position offers an excellent opportunity for someone interested in leading edge
cellular imaging technology (Sigma VP & 3View) to work in a collaborative academic and commercial
environment.
Responsibilities:
 Primary responsibilities include tissue processing, staining and imaging using serial block
face scanning electron microscope (training provided) in accordance with client needs.
 Assist in managing, processing and analysis of 3DEM data.
Other duties may include:
Identifying, developing, and standardizing methods for current and potential 3DEM applications.
Assisting with other projects and analyses when necessary.
Instructing new lab personnel upon completion of the initial employee training period.
Skills and Experience:
 An Associates or Bachelor's degree in material or biological sciences with some experience
in EM related research and/or commercial labs
 Experience in biological and/or material microscopy, including hands-on experience with
sample preparation to include working under a dissecting scope
 Applicants must be dexterous and have excellent eye hand coordination
 Must have good general lab and computer skills. Prior experience with basic image
processing software (eg ImageJ/Recontruct) a plus.
 Knowledge in computer programming (R, C++, Java ) is a positive
 Must be able to work independently and manage timelines to produce deliverables within
deadlines
 Good communication skills and an ability to collaborate with others in cross-functional
teams is a necessity
Eligible candidates can forward your applications/enquiries to aroholt-at-renovoneural.com

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Email: rnichols-at-bcm.edu
Name: Ralph Nichols

Organization: Baylor College of Medicine

Title-Subject: [Filtered] Immuno gold labeling cell culture

Message: Hello Listers,

I hope you all are staying cool! I have a post doc that want to do immuno gold labeling on cell
cultures.He's want to label the cell membranes so spinning them down into a pellet will not do for
his purposes.My questions, if anyone would be so kind as to share some procedures or protocols, (1)
what should he grow his cells on? (2)How to process and embed them with LR White?
You can email off line with your suggestions.

Thanks in advance!

Regards
Ralph

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I would like to thank everyone that took part and assisted us with the
evaluation of our propriatary Blue Platinum EM Stain.

Our tallied results showed that it indeed has potential as a suitabele
replacement for our Uranyl Acetate that we manufacture when there are
either import issues or usage issues with Uranyl Acetate and its effects.

Thank you again
Alex Besenyo PhD

--
INTERNATIONAL BIO-ANALYTICAL INDUSTRIES,INC.

3495 N. Dixie Hwy. Unit # 8
Boca Raton, Florida 33431

Tel: 561-826-0061
Fax: 561-892-8450
Web Site: http://www.ibilabs.com
Email: bioanalytics-at-ibilabs.com

CONFIDENTIALITY NOTICE

This email contains confidential information and may be otherwise
protected by law. Its content should not be disclosed and it should not
be given or copied to anyone other than the person(s) named or referenced
above. If you have received this email in error, please contact the
sender.

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Dear Colleagues,

I am working on a preliminary program for next year's 3DEM GRC, which will take place in Girona, Spain, not far from Barcelona, on June 22-27, 2014. I hereby invite you to send me suggestions for contributions from our community. Any recommendations for speakers for that meeting are most welcome. Please also don't hesitate to nominate yourself, if you think you have exciting new results or developments to report.

With best wishes,

Henning Stahlberg.

Henning Stahlberg, PhD
Prof. for Structural Biology, C-CINA, Biozentrum, University Basel
Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland
http://c-cina.org | Tel. +41-61-387 32 62 | mailto:Henning.Stahlberg-at-unibas.ch

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OK...first...I'm still working with start-up labs with very(!) limited funding and experience.

BUT! even though it would seem I am interested in lower cost solutions I am), I would like to keep the discussion open to all levels (high and low SEM resolution/Thermal, Environmental/Variable Pressure/FE-Cold and Schottky-Thermal, STEM/TSEM...other?)

I am aware of the older more common Low and High Magnification SEM Standards:
a) Low Magnification Standards-grids(various openings per mm(?)
b) High Resolution Standards- Gold on Carbon, Replicas of Optical and UV Gratings.

I am currently using a Gold on Carbon Standard mounted on an aluminum stub(no hole in stub). I have never care much for this standard, but it is very "popular".

What else do you use?

We are probably going to get into the "line-to-line"/Point-to-point/higher and lower contract discussions. What's new??

I realize this might get commercial, but isn't that necessary?(another topic)
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Hi everyone:

I am having a weird problem with my Zeiss LSM 700 confocal microscope and hope you could shed light on it -

When I tried to scan with the lasers off (turning the lasers off via the ZEN software), I can literally see a relatively bright light coming from the objective! The light looks white, which I assume is the combination of the blue, green and red lasers. Although the detected fluorescence signal is 'minimal', it still worries that 1). It might photobleach my sample; and 2). When I scan with one laser, the other lasers might be on as well, so I wouldn't know what exactly I am scanning with!

I was told by Zeiss that this is "normal" because there are no mechanical shutters in the scan head. I have a hard time to believe it and here are my questions:

1). Does anyone experience the same/similar problem?
2). What could be the possible cause?
3). How am I supposed to fix it?

Thank you,

Zhaojie

Zhaojie Zhang, Ph. D.
Director, Jenkins Microscopy Facility
University of Wyoming
Laramie, WY 82071
PHONE: 307-766-3038
FAX: 307-766-5625

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Digital Image Processing: An Algorithmic Introduction using Java, Burger and Bunge

http://www.amazon.com/Digital-Image-Processing-Algorithmic-Introduction/dp/1846283795/ref=sr_1_5?s=books&ie=UTF8&qid=1376334107&sr=1-5&keywords=digital+image+processing

The Image Processing Handbook, Sixth Edition, Russ

http://www.amazon.com/Image-Processing-Handbook-Sixth/dp/1439840458/ref=sr_1_1?s=books&ie=UTF8&qid=1376334213&sr=1-1&keywords=digital+image+processing+russ

My humble opinion - perhaps not with everything you want,

Fred Monson

Frederick C. Monson, PhD
Technical Director
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pennsylvania
Schmucker Science South - Room SSS-024
MailDrop: Geology-Astronomy
750 South Church Street
West Chester, PA, 19383
610-738-0437
fmonson-at-wcupa.edu

HomePage: http://cmirt.wcupa.edu (with link to instrument Scheduler)

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, August 09, 2013 8:24 AM
To: Monson, Frederick

Pawley's "Handbook of Biological Confocal Microscopy" has already been suggested to the OP.

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realname - Nicholas Galvan
Email - nicholas.galvan-at-colorado.edu
ORGANIZATION - Colorado State University EDUCATION - Undergraduate College LOCATION - Fort Collins, Colorado, USA SUBJECT_OF_QUESTION - High Speed Microscopy QUESTION - I am looking for a book that covers the basics of Digital Microscopy, and a book that would be a good follow up to Jerome Mertz'
book Optical Microscopy. I am hoping that book will cover fraunhofer diffraction, fresnel diffraction, radiometry, Fluorescence, and confocal microscopy. Thank you for your time in advanced.

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Title-Subject: [Filtered] LKB knifemaker

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Dear all,

Any thoughts on whether we can replace uranyl stains with this Pt or other
heavy metal stains?

Our institution is on a safety/money-saving drive to replace all potential
sources of radioactivity. Soon we will no longer use 32P for DNA gels (and
no more ethidium bromide either) and I'm asked to get rid of U. acetate.
The radiation licence is very expensive, as is keeping track of the
radiation badges plus disposal of waste, so trying to save money as well
as use safer chemicals.

Interested in others' experiences.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

On 12/08/13 11:22 PM, "bioanalytics-at-ibilabs.com"
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Hi Rosemary,

} Any thoughts on whether we can replace uranyl stains with this Pt or other
} heavy metal stains?

we have extensively tested several alternatives to UA for both manual as well as automated thin section post-staining. A summary of these experiments can be found at

http://bit.ly/VQSmJm

To cut the long story short, in our hands Oolong Tea Extract/lead citrate gave a rather weak stain, even using extended incubation times, and an increased risk of specimen contamination. The sections stained with Platinum blue/lead citrate were very clean and the stain was comparable with UA in density. For interpretation of the micrographs, possible variations in the distribution of the stain have to be taken into account, however.

Negative staining is obviously an entirely different story, so we haven't gotten completely rid of UA to date. Here, I am very interested in the experience of others.

Best,

Guenter

--
Dr. Guenter Resch
Nexperion e.U. - Solutions for Electron Microscopy - www.nexperion.net
Mattiellistrasse 3/17, 1040 Vienna, Austria - Phone +43 664 94 17 210
Registered at Commerical Court Vienna, FN 397677w - VAT Reg. ATU67962234

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We think we do not quite understand the operations manual for the T330A concerning Auto-Focus/Auto-Astigmate. It looks and seems pretty straight forward, but focusing on low contrast "Gold on Carbon"(Al Substrate Stub)Samples seems to be more difficult than I remember on other older Thermal Filament SEMs(35, 840s, etc.). I get the feeling we are confusing the computer system inadvertently. I have been told we can reset/'reboot' the computer by re-starting and avoid! turning the system-key to "Off".
I would like to actually speak to someone that is very familiar with T330A daily operations and common idiosyncrasies of this focus/astigmatism system.
Please email me directly if you have the experience and time for a call.... at your convenience!

Jeff Day
wa5ekh-at-juno.com
____________________________________________________________
30-second trick for a flat belly
This daily 30-second trick BOOSTS your body's #1 fat-burning hormone
http://thirdpartyoffers.juno.com/TGL3131/5209d9cf5ae2359cf57fast01vuc

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Dear Listers,
dear Rosemary,

besides the recent offers on MSA-Listserver for Pt-blue staining and Oolong-Tea-Extract substitute for uranyl acetate
I have posted Dec 7, 2011 on
https://www.researchgate.net/post/TEM_Substitute_reagents_for_Uranyl-Acetate_in_positive_and_negative_staining_of_resin_ultrathin_sections_Dear_Colleagues_I_would_like_to_inform_you_of_an_article_which_recently_was_published_in_Journ2

the following:
(TEM): Substitute reagents for Uranyl-Acetate in positive and negative staining of resin ultrathin sections.

Dear Colleagues,
I would like to inform you of an article which recently was published in Journal of Electron Microscopy(Toyko) which I found to be perhaps of interest also to you Electron Microscopists:

Masamichi Nakakoshi, Hideo Nishioka, and Eisaku Katayama
New versatile staining reagents for biological transmission electron microscopy that substitute for uranyl acetate
Abstract: Aqueous uranyl acetate has been extensively used as a superb staining reagent for transmission electron microscopy of biological materials. However, recent regulation of nuclear fuel material severely restricts its use even for purely scientific purposes. Since uranyl salts are hazardous due to biological toxicity and remaining radioactivity, development of safe and non-radioactive substitutes is greatly anticipated. We examined two lanthanide salts, samarium triacetate and gadolinium triacetate, and found that 1-10% solution of these reagents was safe but still possess excellent capability for staining thin sections of plastic-embedded materials of animal and plant origin. Although post-fixation with osmium tetroxide was essential for high-contrast staining, post-staining with lead citrate could be eliminated if a slow-scan CCD camera is available for observation. These lanthanide salts can also be utilized as good negative-staining reagents to study supramolecular architecture of biological macromolecules. They were not as effective as a fixative of protein assembly, reflecting the non-hazardous nature of the reagents.
Key words: transmission electron microscopy - staining reagent - negative staining - uranyl acetate substitute - samarium triacetate - gadolinium triacetate
J Electron Microsc (Tokyo) 2011 60: 401-407; doi:10.1093/jmicro/dfr084.
http://jmicro.oxfordjournals.org/cgi/content/abstract/60/6/401?etoc
No comment further but: at this point not knowing about prices of the substances mentioned in the article.
Edition of post 12-01-23: It is already known for years that e.g. lanthanum nitrate as a reagent (either added to the fixative or as separate incubation solution [cave: specific recipes to be followed!] has some special effects on retention of elemental substrates in human tissue preparation. Note: a copy of the article by Nakakoshi et al in J Electron Microsc (Tokyo) 2011 60 I own, so if you'd like to get more {info} , please request from w.muss-at-salk.at,

Replies to that post:
Jan 8, 2013
Yaroslav Tsytsyura � Westf�lische Wilhelms-Universit�t M�nster
Thanks for this paper, we have already tried it and now switch to samarium instead of UA in routine protocols. Works both (en block and on grids) well.
Thanks for this paper, we have already tried it and now switch to samarium instead of UA in routine protocols. Works both (en block and on grids) well.

Aug 7, 2013
Stefan Sch�ffberger � Science Services GmbH (Munich, Germany)
Electron Microscopy Sciences just introduced a ready mixed Uranyl Acetate Replacement Stain based on samarium and gadolinium triacetate... just for your information.
Electron Microscopy Sciences just introduced a ready mixed Uranyl Acetate Replacement Stain based on samarium and gadolinium triacetate... just for your information.

Aug 7, 2013
Wolfgang Muss � Paracelsus Medical University Salzburg
Dear all, since I guess this post fits in here too, I would like to point to perhaps another possibility:
non-isotopic (4%) Hafnium chloride in (100%) Methanol used as a contrasting/staining solution for ultrathin sections (SPURRS, 60/80 nm thickness):
(cf: http://onlinelibrary.wiley.com/doi/10.1002/jemt.20964/pdf or for the abstract see:
http://onlinelibrary.wiley.com/doi/10.1002/jemt.20964/abstract).
IKEDA et al, Microscopy Research and Technique Volume 74, Issue 9, pages 825-830, September 2011:
{Enhanced effects of nonisotopic hafnium chloride in methanol as a substitute for uranyl acetate in TEM contrast of ultrastructure of fungal and plant cells.}
Keywords: nonisotope; hafnium chloride; uranyl acetate; ultrastructural contrast; fungus; plant
Abstract: This ultrastructural study showed that nonisotopic methanolic hafnium chloride and aqueous lead solution was an excellent new electron stain for enhancing TEM contrasts of fungal and plant cell structures. The ultrastructural definition provided by the new stain was often superior to that provided by conventional staining with uranyl acetate and lead. Definition of fine ultrastructure was also supported by quantitative data on TEM contrast ratios of organelles and components in fungal and plant cells. In particular, polysaccharides, which were localized in cell walls, glycogen particles, starch grains, and plant Golgi vesicle components, were much more reactive to the new stain than to the conventional one. The new nonisotopic stain is useful for enhancing the contrast of ultrastructure in biological tissues and is a safer alternative to uranyl acetate. Microsc. Res. Tech., 2011. � 2010 Wiley-Liss, Inc. This - I think - is really interesting to EM-ists....
Thanks also to Stefan to tell us that the Sm-Gd-triacetate-staining will be / is commercially available now. Best regards, Wolfgang Cancel Save
Dear all, since I guess this post fits in here too, I would like to point to perhaps another possibility:
non-isotopic (4%) Hafnium chloride in (100%) Methanol used as a contrasting/staing solution for ultrathin sections (SPURRS, 60/80 nm thickness):
(cf: http://onlinelibrary.wiley.com/doi/10.1002/jemt.20964/pdf or for the abstract see:
http://onlinelibrary.wiley.com/doi/10.1002/jemt.20964/abstract).
IKEDA et al, Microscopy Research and Technique Volume 74, Issue 9, pages 825-830, September 2011:
{Enhanced effects of nonisotopic hafnium chloride in methanol as a substitute for uranyl acetate in TEM contrast of ultrastructure of fungal and plant cells.}
Keywords: nonisotope; hafnium chloride; uranyl acetate; ultrastructural contrast; fungus; plant
Abstract: This ultrastructural study showed that nonisotopic methanolic hafnium chloride and aqueous lead solution was an excellent new electron stain for enhancing TEM contrasts of fungal and plant cell structures. The ultrastructural definition provided by the new stain was often superior to that provided by conventional staining with uranyl acetate and lead. Definition of fine ultrastructure was also supported by quantitative data on TEM contrast ratios of organelles and components in fungal and plant cells. In particular, polysaccharides, which were localized in cell walls, glycogen particles, starch grains, and plant Golgi vesicle components, were much more reactive to the new stain than to the conventional one. The new nonisotopic stain is useful for enhancing the contrast of ultrastructure in biological tissues and is a safer alternative to uranyl acetate. Microsc. Res. Tech., 2011. � 2010 Wiley-Liss, Inc. This - I think - is really interesting to EM-ists....
Thanks also to Stefan to tell us that the Sm-Gd-triacetate-staining will be / is commercially available now.
Best regards, Wolfgang

PS: As of my knowledge, also EM-Labs of the RKI (Robert-Koch-Institute, Berlin, Germany) uses/has used the Sm-Gd-stain at least sometime sucessfully. And, last but not least: perhaps the old Bi-(wismut)stain could be another alternative.
Disclaimer: no financial interest, no affiliation to any of the commercial companies mentioned in this text.

Hope this can help for a decision,

best regards and have a happy day everywhere

Wolfgang

OR Dr. phil. Wolfgang Muss*
Head of EM-Lab
Univ. Institute of Pathology, SALK-LKH
(Salzburger Landeskliniken gemeinnuetzige GesmbH, Landeskrankenhaus = Fed. State Gen. Hosp.)
Muellner Hauptstrasse 48
A-5020 SALZBURG AUSTRIA/EUROPE
Secretary of SCUR (Society for Cutaneous Ultrastructure Research)
Member of Works Council SALK-LKH & Central Works Council SALK
Safety Counsellor

Web(German): http://ww.salk.at
Web(German): http://www.pmu.ac.at
Phone work: +43+662+4482+4720
Mobile phone work: +43+662+4482-57704
Fax-No. at work: ++43+662+4482-882 ext (please, only by indicating: "c/o W. Muss")
E-Mail work: W.Muss-at-SALK.at
Mobile-phone private: ++43+676+5 369-456
E-Mail private: wij.muss-at-aon.at
ResearchGate: http://www.researchgate.net/profile/Wolfgang_MUSS/

Von: Rosemary.White-at-csiro.au [mailto:Rosemary.White-at-csiro.au]
Gesendet: Dienstag, 13. August 2013 04:27
An: Mu� Wolfgang
Betreff: [Microscopy] Replacing radioactive stains
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Dear all,

Any thoughts on whether we can replace uranyl stains with this Pt or other
heavy metal stains?

Our institution is on a safety/money-saving drive to replace all potential
sources of radioactivity. Soon we will no longer use 32P for DNA gels (and
no more ethidium bromide either) and I'm asked to get rid of U. acetate.
The radiation licence is very expensive, as is keeping track of the
radiation badges plus disposal of waste, so trying to save money as well
as use safer chemicals.

Interested in others' experiences.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Hello Barbara,
I've make a copy of our LKB Knifemake 7801A manual for you.
You can download it from:
http://www2.biomed.cas.cz/~benada/LKB_knifemaker.pdf

The old cutting wheel can be replaced by carbide glass cutting wheel.
There are a lot of companies selling it, e.g.
http://oaklanddiamondtools.com/carbide-glass-cutting-wheel.php

My best regards

Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR. v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

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Email: j.knowles-at-ucl.ac.uk
Name: Jonathan Knowles

Organization: UCL, London

Title-Subject: [Filtered] Polaron sputter coater problem

Message: Hi

We have an E500 sputter coater and have run into a problem and wondered if anyone can point us in
the right direction.

When you turn it on to pump, the gauge moves slightly but is not registering the vacuum. Any
pointers as to what it is likely to be? The gauge itself? The amp board? The switch? Other?!

thanks

Jonathan

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Email: pevsnerp-at-missouri.edu
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Organization: Dept of Electrical and Comput Engineering, Univ of Missouri

Title-Subject: [Filtered] LKB Type 7801B manual

Message: Does anyone have a manual. I would be pleased to pay for a copy. P

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Email: j.knowles-at-ucl.ac.uk
Name: Jonathan Knowles

Organization: UCL, London

Title-Subject: [Filtered] Polaron sputter coater fixed

Message: Dear All

Many thanks for the help with the above problem. I was overthinking this and (as pointed out by
many) was due to a vacuum leak, which is now fixed.

thanks again to all for the help.

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Postdoctoral Opening on Electron Tomography of Virus-Induced Membrane Rearrangements

A highly motivated individual is sought for a post-doctoral position to use electron microscope tomography in studies of membrane remodeling in HIV and positive-strand RNA virus replication. These studies will integrate advanced, three-dimensional electron tomographic imaging with molecular genetics, biochemistry and other approaches in a collaboration between the Morgridge Institute for Research and the University of Wisconsin � Madison.

Candidates should have a Ph.D. in cell biology, microbiology, biochemistry or a related field and proven experience in electron microscopy. Experience with electron tomography is highly desirable, and experience with cryo-electron microscopy is also valued. Excellent written and oral presentation skills and the ability to work collaboratively with others are expected.
The successful candidate will join a multidisciplinary team including virologists, cell biologists, biochemists and biophysicists in a well-equipped, dynamic and interactive research environment on the campus of the UW-Madison, a world-class academic institution with an international reputation for innovative basic and applied research. Nationally, UW-Madison ranks third among all U.S. universities for research and development expenditures, exceeding $1.0 billion annually. Madison also ranks among the 'best places to live in the US' and post-docs thrive on its diversity of outdoor and cultural opportunities. For more information, please visit the following websites:

http://discovery.wisc.edu/home/morgridge/
http://www.mcardle.wisc.edu/faculty/bio/ahlquist_p.html
http://www.botany.wisc.edu/otegui/welcome.html
http://www.bmolchem.wisc.edu/faculty/audhya.html

Qualified individuals interested in this opportunity are asked to create an online profile and submit a CV and cover letter detailing your current/previous research and names/address of three references as one document at: http://discovery.wisc.edu/home/morgridge/careers/careers.cmsx
Once the online profile has been created, you will receive a request to have your references email their letters of evaluation.

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Email: Martin.Saunders-at-uwa.edu.au
Name: Martin Saunders

Organization: The University of Western Australia

Title-Subject: [Filtered] Research Officer positions available

Message: The Centre for Microscopy, Characterisation and Analysis (CMCA) at The University of
Western Australia (UWA) has two Senior Research Officer positions available. These positions will
support the Centre's microanalysis (microprobe and SEM) and electron microscopy (SEM and TEM)
capabilities respectively. The latter will support new FEI Titan and Verios instruments scheduled
for delivery in early 2014.

The CMCA is a central university infrastructure facility supporting more than 400 researchers across
the physical, biological, medical and geo- sciences. The role of these positions will involve
training researchers and supporting them in their use of the Centre's instruments. The successful
candidates will be encouraged to build collaborative research links with users of the facilities
and, where feasible, to develop their own research activities.

More details of the positions can be found at:

http://external.jobs.uwa.edu.au/cw/en/job/492547/senior-research-officer-electron-beam-microanalysis

and

http://external.jobs.uwa.edu.au/cw/en/job/492545/senior-research-officer-electron-microscopy

Enquiries can be sent to Martin.Saunders-at-uwa.edu.au

Regards,

Martin Saunders,
Acting Director CMCA.

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Email: guosheng.liu-at-usask.ca
Name: Guosheng Liu

Organization: University of Saskatchewan

Title-Subject: [Filtered] SEM service --Philips SEM 505

Message: Hello All,

I'm looking for an experienced Philips SEM505 specialist who can provide a field service/diagnosis
for our nearly 30-years-old SEM, ideally located in western Canada (or in North America). The main
purpose is to fix (if possible), or to evaluate how much (money and time) we need to restore it.

This machine has performed robustly but is appearing some signs of aging and troubles after so many
productive years. We would like to pay for this checkup before determining whether to repair or
decommission.

Any contact info from either satisfied customers or commercial reps are greatly appreciated. You can
reply offline.

Best regards.

Guosheng Liu
U of Saskatchewan
Canada

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Greetings,

The Hooke College of Applied Sciences, located in Westmont, IL, is offering its transmission electron microscopy short course October 1-3, 2013.� In addition to lectures, this course emphasizes hands-on training using state-of-the-art equipment.� For further details and registration information, please see the link below:

http://www.hookecollege.com/courses/course.asp?COURSE_ID=53

Best regards,

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Hello recipe,
Recently we have a problem of methylene blue staining the blank regions (void of tissue) in our 0.5 um semi-thin sections. Our stain recipe is 1% of each methylene blue, azure II and sodium borate, 0.22 um filtered aqueous solution. We generally place the sections on slides and stain on a hot plate set to 5 (very hot) for 1 min, followed by water rinse. In the past we have always had clear epon after staining, with only the tissue taking up the stain. In the past this would routinely happen with toluidine blue or any Spurr's resin only. I have called the company (Pella) and they insist the eponate 12 resins haven't changed, and they have not come across this. Does anyone have a clue? All answers greatly appreciated.

sincerely,
Michael Delannoy

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Title-Subject: [Filtered] MuMetal Shielding for FE-SEM

Message: Has anyone had experience using MuMetal shielding for an FESEM?

- The EMI source is a conduit burried in the slab about 15' from the microscope
- The disipation we need is about 50% at 60 Hz

I'm aware of the active cancelation systems, however we've been asked to look into other options.

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Dow Chemicals
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Confocal Support Specialist - East - LEI000970

Leica Microsystems is a world leader in microscopes and scientific
instruments. Founded as a family business in the nineteenth century, the
company's history was marked by unparalleled innovation on its way to
becoming a global enterprise.

Its historically close cooperation with the scientific community is the
key to Leica Microsystems' tradition of innovation, which draws on users'
ideas and creates solutions tailored to their requirements. At the global
level, Leica Microsystems is organized in three divisions, all of which
are among the leaders in their respective fields: the Life Science
Division, Industry Division, and Medical Division.

Job Summary:

The purpose of this position is to provide technical sales support for all
Confocal products with the exception of the SPE and LSI. Support will be
provided to Leica's Life Science Research customers, Confocal Applications
Specialist (CAS) in all facets of their selling activities. This includes,
but is not limited to, visiting reps and customers in the field,
conducting product demonstrations, training, participating in local and
national exhibitions, handling phone inquiries, solving applications
problems, providing customer feedback, and assisting the Area Sales
Managers in their motivational activities and assessment of the CAS sales
performance.

. Ensures that customers are satisfied! Also, must be capable of working
effectively with Sales Managers, PLT members, Customer Service Technicians
and Representatives, Engineers, Accounting, and Manufacturing to do
whatever is necessary to satisfy customers' needs.
. Perform customer demonstrations
. Conducts technical product training and provides sales tools to help
Leica sales reps become more effective in selling products. Participates
in all activities (Travel in the field with sales reps , attend industry
meetings, conferences, local and national exhibitions, conduct product
seminars, etc.) that will enhance the awareness, acceptance, and
eventually the sale of Leica Confocal products.
. Assists in meeting sales goals for each of the Confocal sales reps in
his/her assigned territory and provides positive motivation and support to
help sales reps meet their goals.
. Works with Sales Management to develop strategies designed to increase
sales of Leica products in assigned territories. Specifically, he/she
recommends product, pricing, distribution, advertising and sales promotion
strategies.

Requirements
. Prefer masters or PhD or equivalent experience in life science
field
. Experience in working with Confocal microscopy products is a
must
. Sales experience in selling products of a technical nature is
desirable
. In-depth knowledge of the microscopy market is also highly
desirable
. A minimum of two years related microscopy experience in a
laboratory preferably a core facility is essential
. Experience with Leica Confocals is desirable.
. Valid driver's license
. Must have valid credit card and be able to pay for reimbursable
expenses.
. Must live in territory

Danaher Corporation Overview
Danaher Corporation is one of the most remarkable success stories in the
Fortune 500. In addition to Leica, Danaher owns some of the world's
leading industrial brands - products that span the most demanding
applications in the world. Currently over 50 companies and 60,000
employees around the globe are working together challenging each other to
take this success even further. Our joint quest does not only increase
chance of success, it also fosters an environment in which a global career
path is encouraged and limited only by your talent, capacity and
ambitions.

Organization
Leica

Job Function
Sales

Primary Location
North America-United States-DC-Washington

Schedule
Full-time

Please use this link to apply:
https://danaher.taleo.net/careersection/external/jobdetail.ftl?lang=en&job
=lei000970

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Hi everybody,

I'm looking for a simple estimation that I can't find in any textbook I've looked in, nor on the internet.
I wonder if anybody can help me.

If I put a diffraction grating with spacing 1 Angstrom into the specimen holder and record an image of it.
At what distance from the optical axis will the corresponding diffraction spot (for 1 Angstrom spacing)
appear in the back focal plane of the objective lens.

Assume a TEM with 200 kV and a twin lens, one that's typically used for protein structure determination.

Thanks,

Philip

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Philip

You simply need to use Bragg's law to calculate the scattering angle.

Here is a simple table of Diffraction/Bragg Angles (in milliradians)
as a function of D-spacing and Electron Beam Energy (80-300kV)

The actual distance you measure in your DP will depend upon your
camera length. Also remember that the anglular distance from the optic
axis to the Bragg spot is twice the Bragg Angle. Just refer
to any standard TEM text covering diffraction for the details.

For 1 Angstrom d-spacing at 200 keV the Bragg angle is 12.54 mR

d 80 kV 100 kV 200 kV 300 kV
-(�)-----------------(mR)-------------

0.5 41.75 37.00 25.07 19.69
0.6 34.79 30.83 20.89 16.41
0.7 29.82 26.43 17.91 14.06
0.8 26.09 23.13 15.67 12.31
0.9 23.19 20.56 13.93 10.94
1 20.88 18.50 12.54 9.85
1.1 18.98 16.82 11.40 8.95
1.2 17.40 15.42 10.45 8.20
1.3 16.06 14.23 9.64 7.57
1.4 14.91 13.21 8.95 7.03
1.5 13.92 12.33 8.36 6.56
1.6 13.05 11.56 7.83 6.15
1.7 12.28 10.88 7.37 5.79
1.8 11.60 10.28 6.96 5.47
1.9 10.99 9.74 6.60 5.18
2 10.44 9.25 6.27 4.92
2.1 9.94 8.81 5.97 4.69
2.2 9.49 8.41 5.70 4.48
2.3 9.08 8.04 5.45 4.28
2.4 8.70 7.71 5.22 4.10
2.5 8.35 7.40 5.01 3.94
2.6 8.03 7.12 4.82 3.79
2.7 7.73 6.85 4.64 3.65
2.8 7.46 6.61 4.48 3.52
2.9 7.20 6.38 4.32 3.39
3 6.96 6.17 4.18 3.28
3.1 6.73 5.97 4.04 3.18
3.2 6.52 5.78 3.92 3.08
3.3 6.33 5.61 3.80 2.98
3.4 6.14 5.44 3.69 2.90
3.5 5.96 5.29 3.58 2.81
3.6 5.80 5.14 3.48 2.73
3.7 5.64 5.00 3.39 2.66
3.8 5.49 4.87 3.30 2.59
3.9 5.35 4.74 3.21 2.52
4 5.22 4.63 3.13 2.46
4.1 5.09 4.51 3.06 2.40
4.2 4.97 4.40 2.98 2.34
4.3 4.85 4.30 2.92 2.29
4.4 4.74 4.20 2.85 2.24
4.5 4.64 4.11 2.79 2.19
4.6 4.54 4.02 2.73 2.14
4.7 4.44 3.94 2.67 2.09
4.8 4.35 3.85 2.61 2.05
4.9 4.26 3.78 2.56 2.01
5 4.18 3.70 2.51 1.97
6 3.48 3.08 2.09 1.64
7 2.98 2.64 1.79 1.41
8 2.61 2.31 1.57 1.23
9 2.32 2.06 1.39 1.09
10 2.09 1.85 1.25 0.98

And a simple table of Energy to Wavelength Conversion

Eo(kV) Beta (v/c) To(keV) Lambda(�)
10 0.194985588 9.713933639 0.122006809
20 0.271865885 18.88424091 0.085858423
30 0.328376144 27.5507422 0.06977036
40 0.374059778 35.74967755 0.0601388
50 0.412686137 43.51408608 0.053539219
60 0.446224575 50.87413939 0.048648835
70 0.475865846 57.85743509 0.044834105
80 0.502398843 64.4892559 0.041748422
90 0.526380132 70.7927988 0.039184029
100 0.548220817 76.78937814 0.037007783
120 0.586670979 87.93855317 0.033487213
140 0.619564277 98.0760223 0.030736068
160 0.648106642 107.3205897 0.028507488
180 0.673148202 115.7741193 0.026653306
200 0.695314426 123.5243525 0.025078437
250 0.741018616 140.2969866 0.021986933
300 0.776525452 154.0641159 0.01968915

Nestor
Your Friendly Neighborhood SysOp

On 8/23/13 9:18 AM, Philip.Koeck-at-ki.se wrote:
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} I wonder if anybody can help me.
}
} If I put a diffraction grating with spacing 1 Angstrom into the specimen holder and record an image of it.
} At what distance from the optical axis will the corresponding diffraction spot (for 1 Angstrom spacing)
} appear in the back focal plane of the objective lens.
}
} Assume a TEM with 200 kV and a twin lens, one that's typically used for protein structure determination.
}
} Thanks,
}
} Philip
}
}
} ==============================Original Headers==============================
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Hi Philip,

To follow up on Nestor's response...

You can put a standard sample (such as an Al film) into your
microscope. The d-spacings of Al are well known. Hence, using Bragg's
Law, you can calculate the diffraction angles of the rings. You also
have several objective apertures in your microscope with known
diameters. If you put an aperture in over the diffraction pattern, you
can compare the radii and get the absolute distance from the optic axis
of the rings/spots at the Back Focal Plane. Also, knowing the
diffraction angles and the objective aperture diameters, by simple
geometry you can get a pretty good measure of the focal length of the
objective lens.

The distance from the optic axis at the viewing screen is, as Nestor
said, dependent on the camera length. Note that the true camera length
may be somewhat different from the microscope's indicated camera length.

Cheers,
Henk

At 8/23/2013 10:19 AM, Philip.Koeck-at-ki.se wrote:
}
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} Hi everybody,
}
} I'm looking for a simple estimation that I can't find in any textbook I've looked in, nor on the internet.
} I wonder if anybody can help me.
}
} If I put a diffraction grating with spacing 1 Angstrom into the specimen holder and record an image of it.
} At what distance from the optical axis will the corresponding diffraction spot (for 1 Angstrom spacing)
} appear in the back focal plane of the objective lens.
}
} Assume a TEM with 200 kV and a twin lens, one that's typically used for protein structure determination.
}
} Thanks,
}
} Philip
}
}
} ==============================Original Headers==============================
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--
Untitled Document

Hendrik O. Colijn
*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*
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Hi Everybody

I really appreciate your information regarding my question:

I am working with samples of muscle tissue where I believe subset of
individual fibers have membranes holes due to use-and-misuse damage.
Also there are subset of fibers which are at early stages of necrosis.

These samples are fixed with paraformaldehyde (4%) and will be
processed through dehydration and LR white infiltration to be embedded
in LR white blocks for subsequent sectioning.

Before embedding, I want to discern fibers that have holes from
use-and-misuse, or necrosis. I am investigating different ways of
classifying these two different groups of fiber by the size of the
larges holes in their membrane. I am finding an upper bound of the
sizes of the membrane holes that are created by use-and-misuse or
necrosis. I am doing this by finding the exclusion sizes of
macromolecules capable of diffusing across plasmalemma into sarcoplasm
of damaged and necrotic fibers in the muscle tissue, with using
dextran molecules that have different sizes.

Does anybody know the size of pores that are created from PFA fixation
so I can use macromolecules that are larger than those pore sizes to
avoid assaying those pore sizes- I know that there are cases that
antibodies penetrate into the cytoplasm after just PFA fixation
without permeabilization with detergents, which implies that these
pore sizes are larger than the size of some antibodies.

Does anybody also know the range of hole sizes from early stages of necrosis?

What are good markers for necrosis that can be detected with antibodies?

I appreciate any information or references; thank you in advance.

Mariam Ghochani
Doctoral Research Training Fellow
Laboratory of Cellular and Molecular Biophysics
National Institute of Child Health and Human Development
9000 Rockville Pike, Bldg 10, Room 10D18
Bethesda, MD 20892
Tel: (301)435-3069 / (818)216-4474
E-mail: mariamghochani-at-gmail.com

==============================Original Headers==============================
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Email: mariamghochani-at-gmail.com
Name: Mariam Ghochani

Organization: NIH, NICHD

Title-Subject: [Filtered] size of pores from paraformaldehyde permeabilization

Message: Hi Everybody

I really appreciate your information regarding my question:

I am working with samples of muscle tissue where I believe subset of individual fibers have
membranes holes due to use-and-misuse damage. Also there are subset of fibers which are at early
stages of necrosis.

These samples are fixed with paraformaldehyde (4%) and will be processed through dehydration and LR
white infiltration to be embedded in LR white blocks for subsequent sectioning.

Before embedding, I want to discern fibers that have holes from use-and-misuse, and necrosis. I am
investigating different ways of assaying these two different groups of fiber by finding the upper
bound of their membrane hole sizes that are created by use-and-misuse or necrosis. I am doing this
by finding the exclusion sizes of macromolecules capable of diffusing across plasmalemma into
sarcoplasm of these fibers in the muscle tissue, with using different dextran sizes.

Does anybody know the size of pores that are created from PFA fixation so I can use macromolecules
that are larger than those pore sizes so I avoid assaying for those pore sizes- I know that there
are cases that antibodies penetrate into the cytoplasm after just PFA fixation without
permeabilization with detergents, which implies that these pore sizes are larger than the size of
some antibodies.

Does anybody also know the range of hole sizes from early stages of necrosis?

What are good markers for necrosis that can be detected with antibodies?

I appreciate any information or references; thank you in advance.

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Listserver Email Form V - 20120416
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Email: crnl_srbu-at-yahoo.com
Name: Corneliu Sarbu

Organization: INCDFM Bucharest

Title-Subject: [Filtered] reply to "electron "diffraction" & my own question

Message:
Hi, Philip,

on what purpose do you want to do that correlation, i.e. between the 1 angstroem distance in your
real space of sample and the g-vectors, the "distance" of any diffraction spot ? Remember, the
correlation between a limited area of the direct space of your sample and the diffraction entities
can be done by SAED, and the SA aperture is placed much lower in the column than the sample holder,
i.e. the level of your real space sample. Besides, the correlation between the real space sample and
the diffraction "product" is done via a Fourier Transform, and therefore there is no linear correlation.

I have a question of mine, addressed to Nestor: how can be used the new kind of bars which are
printed on any diffractograms and show a number of 1/nm units, for the direct measurement, on that
"plate", of the reciprocal space vector lengths ? What is the meaning of that unit, namely [1/nm] ?
I have difficulties in getting familiar with the use of it, so that I still use the accel. voltage +
camera length to "measure" the g-vectors, but that is a more laborious effort.

Corneliu Sarbu

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Email: duane.harland-at-agresearch.co.nz
Name: Duane Harland

Organization: AgResearch, New Zealand

Title-Subject: [Filtered] Calibration grids

Message: Greetings fellow microscopists

The recent emails on electron diffraction got me thinking.

Overview:
Can anyone let me know what substrates they have found successful for calibrating the scale-bar of
cameras at high magnifications?

Background:
I have a 100kV TEM in which the standard etched line and cross-gratings are fine for calibrating our
Megaview 3 camera in the range 100x to 70kx, but higher than that it gets less precise as
inconsistencies in the gratings become clearer and the number of lines in view become fewer (down to
1 line at 140kx).

Problems:
1. More projects are working entirely at high mag looking at protein assemblies and fibrils etc. And
accurate measurements are important.
2. I have a 300kV machine that has just been fixed after an extended downtime and high-mag
tomography and negative-stain protein complex work will require really good camera calibration.

Questions:
1. Are there better gratings out there?
2. Do people use standard samples (e.g., negative stained TMV or like)?
3. Am I barking up the wrong tree and there is some diffraction method + sample that works better
than direct measurement?

I really value any experiences,
Duane Harland, AgResearch, Lincoln, New Zealand

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Hi Duane

For many years prior to the microcircuit industry becoming involved in
calibration standards we used the crystal lattice of suitable materials as a
high magnification standard. Copper phhalocyanine (1nm), potassium
chloroplatinate (0.56nm) and graphitised carbon (0.34nm) are specimens
available from the usual EM accessory providers and with the capabilities of
modern instruments easily identifiable.

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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Email: duane.harland-at-agresearch.co.nz
Name: Duane Harland

Organization: AgResearch, New Zealand

Title-Subject: [Filtered] Calibration grids

Message: Greetings fellow microscopists

The recent emails on electron diffraction got me thinking.

Overview:
Can anyone let me know what substrates they have found successful for
calibrating the scale-bar of cameras at high magnifications?

Background:
I have a 100kV TEM in which the standard etched line and cross-gratings are
fine for calibrating our Megaview 3 camera in the range 100x to 70kx, but
higher than that it gets less precise as inconsistencies in the gratings
become clearer and the number of lines in view become fewer (down to
1 line at 140kx).

Problems:
1. More projects are working entirely at high mag looking at protein
assemblies and fibrils etc. And accurate measurements are important.
2. I have a 300kV machine that has just been fixed after an extended
downtime and high-mag tomography and negative-stain protein complex work
will require really good camera calibration.

Questions:
1. Are there better gratings out there?
2. Do people use standard samples (e.g., negative stained TMV or like)?
3. Am I barking up the wrong tree and there is some diffraction method +
sample that works better than direct measurement?

I really value any experiences,
Duane Harland, AgResearch, Lincoln, New Zealand

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Thank you all very much for the answers you sent. I just picked one out to reply to.
Thank you , Nestor, for the tables.

The reason I asked is that I want to know what spatial frequencies are removed by an objective aperture
when recording high resolution (phase contrast) images.
I wasn't sure whether the objective lens settings change when one switches between imaging and diffraction.
Therefore the strange way of asking the question.
Secondly I wasn't sure whether one can use the simple formulas given that the lens is not a thin lens.
I don't know the position of the specimen with respect to the focal planes or the principal planes either.

In any case everybody seems to point me to a simple calculation.
For example for 200 kV and 1 Angstrom resolution cutoff I can take twice the angle in Nestor's table,
resulting in angle= 25 mrad.
I can also get that from Lamda = d sin(angle), if I regard the specimen as a two dimensional grating.

The next step is to use the focal length (not the distance between specimen and focal plane) as camera length,
giving a distance of the spot from the optical axis R = 72.5 micrometers for 2.9 mm focal length (f).
The formula is f * tan(angle) = R.
Most answers seem to agree about using f.

Please tell me if I got anything wrong.

Philip

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X-from: Wim Hagen [mailto:wim.hagen-at-embl.de]
Sent: 24 August 2013 14:05
To: Philip K�ck
Cc: microscopy-at-microscopy.com; 3dem-at-ncmir.ucsd.edu

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Title-Subject: [Filtered] JEOL 6300F Bakeout procedure

Message: I have recently purchased a used Jeol 6300F SEM. I am working on quite a budget, and thus,
having JEOL come in to do the bakeout procedure on the instrument or the alignment procedure is so
far out of the question. I was hoping that someone on this list server would have a bakeout
procedure in some sort of format that they could send to me and perhaps even a good alignment
procedure?

I have the standard Jeol instructions, but surprisingly they do not give much info about the bakeout
procedure. If anyone happened to have anything extra and were willing to share, I would be very
grateful!

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Philip

Since you have clarified your question further I should
also point out that the diffraction aperture
is not always the limiting factor in phase contrast
imaging particularly at the 1 Angstrom level that you
seem to be interested in.

When performing HR imaging experiments in this regime,
one generally does not use any apertures.

In HREM , you will need to invoke and understand the limits
created by the aberrations (Cs & Cc) of your Imaging Lenses.
Of these, the Objective Lens is the most critical.

For high resolution work (nominally anything below
about 0.3 nm) you will need to understand the Phase Contrast
Transfer Function of your instrument.

I recommend that you should pickup a good TEM textbook
or review article which discusses High Resolution Phase Contrast Imaging.

The book by Carter and Williams will be a good staring point.

There are also several software programs available
on the net which will calculate an approximate PCTF for you.

See for example

http://www.maxsidorov.com/ctfexplorer/webhelp/background.htm

Just to give you a ball park value, for a 200 kV
FEI FEGTEM (Cs ~ 2 mm) without aberration correctors,
the spatial frequency cutoff is in the vicinity
of 0.2-0.3 nm.

Cheers,

Nestor
Your Friendly Neighborhood SysOp.

On 8/26/13 7:28 AM, Philip.Koeck-at-ki.se wrote:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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}
} Thank you all very much for the answers you sent. I just picked one out to reply to.
} Thank you , Nestor, for the tables.
}
} The reason I asked is that I want to know what spatial frequencies are removed by an objective aperture
} when recording high resolution (phase contrast) images.
} I wasn't sure whether the objective lens settings change when one switches between imaging and diffraction.
} Therefore the strange way of asking the question.
} Secondly I wasn't sure whether one can use the simple formulas given that the lens is not a thin lens.
} I don't know the position of the specimen with respect to the focal planes or the principal planes either.
}
} In any case everybody seems to point me to a simple calculation.
} For example for 200 kV and 1 Angstrom resolution cutoff I can take twice the angle in Nestor's table,
} resulting in angle= 25 mrad.
} I can also get that from Lamda = d sin(angle), if I regard the specimen as a two dimensional grating.
}
} The next step is to use the focal length (not the distance between specimen and focal plane) as camera length,
} giving a distance of the spot from the optical axis R = 72.5 micrometers for 2.9 mm focal length (f).
} The formula is f * tan(angle) = R.
} Most answers seem to agree about using f.
}
} Please tell me if I got anything wrong.
}
} Philip
}
}
} -----Original Message-----
} X-from: Wim Hagen [mailto:wim.hagen-at-embl.de]
} Sent: 24 August 2013 14:05
} To: Philip K�ck
} Cc: microscopy-at-microscopy.com; 3dem-at-ncmir.ucsd.edu
} Subject: Re: [3dem] electron diffraction
}
} Hi Philip,
}
} A simple estimation:
}
} Angular distance in back focal plane [rad] = 2 * Bragg angle [rad] = Lambda (relativistic wavelength) [m] / d (sample spacing) [m] = R (distance) [m] / F (focal length) [m]
}
} All CM/Tecnai Twin systems have a focal length of ~2.9 mm. For 200KV, a 0.1 nm spacing gives ~25 mrad so ~72.5 micrometers distance from optical axis in back focal plane.
}
} Put in the radius of your objective apertures for R and you can calculate where your objective apertures cut off resolution for each KV.
}
} Best,
}
} Wim
}
}
} On Aug 23, 2013, at 4:18 PM, Philip K�ck wrote:
}
} } Hi everybody,
} }
} } I'm looking for a simple estimation that I can't find in any textbook I've looked in, nor on the internet.
} } I wonder if anybody can help me.
} }
} } If I put a diffraction grating with spacing 1 Angstrom into the specimen holder and record an image of it.
} } At what distance from the optical axis will the corresponding
} } diffraction spot (for 1 Angstrom spacing) appear in the back focal plane of the objective lens.
} }
} } Assume a TEM with 200 kV and a twin lens, one that's typically used for protein structure determination.
} }
} } Thanks,
} }
} } Philip
} } _______________________________________________
} } 3dem mailing list
} } 3dem-at-ncmir.ucsd.edu
} } https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem
}
} -------------------------------------------------------------------
} Wim Hagen
} Senior Engineer Electron Microscopy
} European Molecular Biology Laboratory (EMBL)
} Heidelberg, Germany
} --------------------------------------------------------------------
}
}
}
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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Lowicryl K4M Embedment

Message: We have been having challenges with FluoroNanogold labeling of a very stubborn antigen on
the surface of lipid droplets in murine brown adipose tissue. We use a rabbit polyclonal primary
antibody and the goat anti-rabbit Fab' FNG probe. We also have now tried embedment in Lowicryl K4M
with high energy UV polymerization and using benzoin ethyl ether in the resin. We use dry ice chips
for an O.N. polymerization to lower the temperature to hopefully preserve the antigencity.
Unfortunately the chips evaporate by the time the next morning rolls around. The lamp distance is
approximately 7 inches. The whole set up is tented in Al foil and the styrofoam box is also lined as
well.
Our first attempt of this at RT and UV polymerization failed to detect the antigen and the
ultrastructure was not good. The only great thing was that the lipid droplets retained their slate
grey tone, indicating that lipids were preserved. LR White tends to dissolve our lipids, but in
other tissues, antigenicity is well preserved.
Thanks, experts, for your help!
Best, Vickie

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Duane,

There really is only one magnification standard to use for the TEM and that
is the Mag-I-Cal sample. It can be used at all magnifications from the
lowest to the highest on any TEM. In addition, it can calibrate the camera
constant and image rotation.

When you use other standards for the different mag ranges, you not only have
the problem that you state, but often the results don't agree at the overlap
range. In addition, some of them are beam sensitive.

-Scott Walck

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Email: duane.harland-at-agresearch.co.nz
Name: Duane Harland

Organization: AgResearch, New Zealand

Title-Subject: [Filtered] Calibration grids

Message: Greetings fellow microscopists

The recent emails on electron diffraction got me thinking.

Overview:
Can anyone let me know what substrates they have found successful for
calibrating the scale-bar of cameras at high magnifications?

Background:
I have a 100kV TEM in which the standard etched line and cross-gratings are
fine for calibrating our Megaview 3 camera in the range 100x to 70kx, but
higher than that it gets less precise as inconsistencies in the gratings
become clearer and the number of lines in view become fewer (down to
1 line at 140kx).

Problems:
1. More projects are working entirely at high mag looking at protein
assemblies and fibrils etc. And accurate measurements are important.
2. I have a 300kV machine that has just been fixed after an extended
downtime and high-mag tomography and negative-stain protein complex work
will require really good camera calibration.

Questions:
1. Are there better gratings out there?
2. Do people use standard samples (e.g., negative stained TMV or like)?
3. Am I barking up the wrong tree and there is some diffraction method +
sample that works better than direct measurement?

I really value any experiences,
Duane Harland, AgResearch, Lincoln, New Zealand

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Hi Nestor,

I assume you are talking about the first zero of the CTF in (extended) Scherzer defocus.

In structural biology we always work at high underfocus, so we have to go far beyond the first
zero by some sort of CTF-correction and combining images with different defocus.

The resolution range I'm thinking about is between 10 Angstrom and about 2 Angstrom so I think
the size of the aperture should be important though maybe not in a completely straight forward
way as Marin pointed out.

I'm not sure if anybody has managed to get useful images of biological molecules without an aperture.
Might be worth trying.

Philip

-----Original Message-----
X-from: zaluzec-at-aaem.amc.anl.gov [mailto:zaluzec-at-aaem.amc.anl.gov]
Sent: 26 August 2013 17:28
To: Philip K�ck

Philip

Since you have clarified your question further I should also point out that the diffraction aperture is not always the limiting factor in phase contrast imaging particularly at the 1 Angstrom level that you seem to be interested in.

When performing HR imaging experiments in this regime, one generally does not use any apertures.

In HREM , you will need to invoke and understand the limits created by the aberrations (Cs & Cc) of your Imaging Lenses.
Of these, the Objective Lens is the most critical.

For high resolution work (nominally anything below about 0.3 nm) you will need to understand the Phase Contrast Transfer Function of your instrument.

I recommend that you should pickup a good TEM textbook or review article which discusses High Resolution Phase Contrast Imaging.

The book by Carter and Williams will be a good staring point.

There are also several software programs available on the net which will calculate an approximate PCTF for you.

See for example

http://www.maxsidorov.com/ctfexplorer/webhelp/background.htm

Just to give you a ball park value, for a 200 kV FEI FEGTEM (Cs ~ 2 mm) without aberration correctors, the spatial frequency cutoff is in the vicinity of 0.2-0.3 nm.

Cheers,

Nestor
Your Friendly Neighborhood SysOp.

On 8/26/13 7:28 AM, Philip.Koeck-at-ki.se wrote:
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}
} Thank you all very much for the answers you sent. I just picked one out to reply to.
} Thank you , Nestor, for the tables.
}
} The reason I asked is that I want to know what spatial frequencies are
} removed by an objective aperture when recording high resolution (phase contrast) images.
} I wasn't sure whether the objective lens settings change when one switches between imaging and diffraction.
} Therefore the strange way of asking the question.
} Secondly I wasn't sure whether one can use the simple formulas given that the lens is not a thin lens.
} I don't know the position of the specimen with respect to the focal planes or the principal planes either.
}
} In any case everybody seems to point me to a simple calculation.
} For example for 200 kV and 1 Angstrom resolution cutoff I can take
} twice the angle in Nestor's table, resulting in angle= 25 mrad.
} I can also get that from Lamda = d sin(angle), if I regard the specimen as a two dimensional grating.
}
} The next step is to use the focal length (not the distance between
} specimen and focal plane) as camera length, giving a distance of the spot from the optical axis R = 72.5 micrometers for 2.9 mm focal length (f).
} The formula is f * tan(angle) = R.
} Most answers seem to agree about using f.
}
} Please tell me if I got anything wrong.
}
} Philip
}
}
} -----Original Message-----
} X-from: Wim Hagen [mailto:wim.hagen-at-embl.de]
} Sent: 24 August 2013 14:05
} To: Philip K�ck
} Cc: microscopy-at-microscopy.com; 3dem-at-ncmir.ucsd.edu
} Subject: Re: [3dem] electron diffraction
}
} Hi Philip,
}
} A simple estimation:
}
} Angular distance in back focal plane [rad] = 2 * Bragg angle [rad] =
} Lambda (relativistic wavelength) [m] / d (sample spacing) [m] = R
} (distance) [m] / F (focal length) [m]
}
} All CM/Tecnai Twin systems have a focal length of ~2.9 mm. For 200KV, a 0.1 nm spacing gives ~25 mrad so ~72.5 micrometers distance from optical axis in back focal plane.
}
} Put in the radius of your objective apertures for R and you can calculate where your objective apertures cut off resolution for each KV.
}
} Best,
}
} Wim
}
}
} On Aug 23, 2013, at 4:18 PM, Philip K�ck wrote:
}
} } Hi everybody,
} }
} } I'm looking for a simple estimation that I can't find in any textbook I've looked in, nor on the internet.
} } I wonder if anybody can help me.
} }
} } If I put a diffraction grating with spacing 1 Angstrom into the specimen holder and record an image of it.
} } At what distance from the optical axis will the corresponding
} } diffraction spot (for 1 Angstrom spacing) appear in the back focal plane of the objective lens.
} }
} } Assume a TEM with 200 kV and a twin lens, one that's typically used for protein structure determination.
} }
} } Thanks,
} }
} } Philip
} } _______________________________________________
} } 3dem mailing list
} } 3dem-at-ncmir.ucsd.edu
} } https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem
}
} -------------------------------------------------------------------
} Wim Hagen
} Senior Engineer Electron Microscopy
} European Molecular Biology Laboratory (EMBL)
} Heidelberg, Germany
} --------------------------------------------------------------------
}
}
}
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Hi Vicki,

Do you have access to a -20 walk-in freezer, or even the freezer compartment of a standard fridge? Similar to your system, we used a foil-lined cardboard box but with a "black light" bulb like those used to illuminate posters back in the '60s. We've used dry ice or liquid nitrogen to contribute an atmosphere allowing LRWhite and K4M to polymerize with good success. Also, molecular probes now distributes a secondary conjugate with a flourophore and -10 nm gold that works exceptionally well.

Good luck,

Doug

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Email: vakimler-at-med.wayne.edu
Name: Vickie Kimler

Organization: Wayne State University School of Medicine

Title-Subject: [Filtered] Lowicryl K4M Embedment

Message: We have been having challenges with FluoroNanogold labeling of a very stubborn antigen on the surface of lipid droplets in murine brown adipose tissue. We use a rabbit polyclonal primary antibody and the goat anti-rabbit Fab' FNG probe. We also have now tried embedment in Lowicryl K4M with high energy UV polymerization and using benzoin ethyl ether in the resin. We use dry ice chips for an O.N. polymerization to lower the temperature to hopefully preserve the antigencity.
Unfortunately the chips evaporate by the time the next morning rolls around. The lamp distance is approximately 7 inches. The whole set up is tented in Al foil and the styrofoam box is also lined as well.
Our first attempt of this at RT and UV polymerization failed to detect the antigen and the ultrastructure was not good. The only great thing was that the lipid droplets retained their slate grey tone, indicating that lipids were preserved. LR White tends to dissolve our lipids, but in other tissues, antigenicity is well preserved.
Thanks, experts, for your help!
Best, Vickie

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Hello Ralph,

Depending on which side of the membrane your antigen resides, immunolabeling cell membranes may be as simple as growing the cultures on thermanox (permanox) coverslips (you can get them from EMS among other sources), then placing the cultures directly on a drop of diluted primary antibody then rinsing, then on secondary antibody-gold conjugate, rinsing, then fixation and embedding in epoxy. You can embed the coverslip in a polypropylene cap (Wheaton "snap caps"). Thermanox coverslips are not degraded by PO or epoxy, but you may have a bit of trouble removing the embedded cells from the epoxy. We trim the block, then dip just the embedded end into liquid nitrogen for a second or two, then pop the coverslip away using a sharp probe under a stereoscope. Now you have cells exposed at the surface. This works for cultures embedded in LRWhite too, should you need to label your cultures after sectioning.

Best of luck,

Doug

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Email: rnichols-at-bcm.edu
Name: Ralph Nichols

Organization: Baylor College of Medicine

Title-Subject: [Filtered] Immuno gold labeling cell culture

Message: Hello Listers,

I hope you all are staying cool! I have a post doc that want to do immuno gold labeling on cell cultures.He's want to label the cell membranes so spinning them down into a pellet will not do for his purposes.My questions, if anyone would be so kind as to share some procedures or protocols, (1) what should he grow his cells on? (2)How to process and embed them with LR White?
You can email off line with your suggestions.

Thanks in advance!

Regards
Ralph

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realname - Heather Eberhardt
Email - heather.eberhardt-at-frx.com
ORGANIZATION - Forest Research Institute
EDUCATION - Graduate College
LOCATION - Commack, NY, USA
SUBJECT_OF_QUESTION - Sputter Coater Recommendations
QUESTION - Hi folks,
I'm in need of recommendations (including both for and against)
regarding sputter coaters and their vendors. The application is SEM of
pharmaceutical materials. We have one now but it seems to be a complete
and total lemon as it lasts on average 3 weeks between needing
repairs... Vendors are welcome to contact me off list.

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Email: robyn.leidel-at-utsouthwestern.edu
Name: Robyn

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Title-Subject: [Filtered] Glow Discharge

Message: Is there any one that can SIMPLY explain theoretically what glow discharge is, how it works
and the relationship between current and polarity? Please?...

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Email: amit.welcomes.u-at-gmail.com
Name: Amit Gupta

Organization: IISc

Title-Subject: [Filtered] RE: electron diffraction limits and phase contrast

Message: I am not sure whether this article will fill your needs, but any way a good
one to look at academically. Link of corresponding paper is provided in
article itself. Its about limits of electron microscopy in aberration
corrected microscopes.

http://physics.aps.org/articles/v6/82

Regards
Amit Gupta

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Email: zhangyc-at-gmail.com
Name: Yucheng Zhang

Title-Subject: [Filtered] TEM HT problem

Message: We have recently acquired a second-handed JEOL-2010 TEM microscope. Unfortunately, some
prob-lems seem to occur to the high tension. The current emission quickly goes up to an abnormally
large value when the acceleration voltage is slowly increased to a certain value (e.g. 50 kV). The
HT tank also makes noise (sounds likes discharge). We have tried to manually increase the voltage
extremly slow (1kV/hour) to check if it will stabilize, but it fails to work. Any advice on the
problem would be much appreciated.
Thanks in advance.

Yucheng

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Hello Heather,
ARDL bought a Cressington Sputter Coater, model 108. It was the cheapest we could find and it works great. I recommend buying the Pt target as interferes less with EDS spectrums with the exception of P.

We weren't interested in film thickness measurements, or refrigerated stages. We just wanted a nice conductive film of metal. I also use Ar as my carrier gas for smaller metal grain size.

We started with an Au target, I was afraid if I asked for a Pt target in addition to the gold it would have hexed the deal. When the gold wore out, I bought a Pt target. The change over was easy.

Get the complete set up with vacuum connections, we pieced it together to save money, but we ended up buying all the parts anyway.

Let me suggest that you don't use the vacuum system to evaporated solvent from mounting your samples. I know some people do this, I worry about containing the pump oil and creating problems later.

Stay safe...............

Frank

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Tuesday, August 27, 2013 4:26 PM
To: Frank Karl

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realname - Heather Eberhardt
Email - heather.eberhardt-at-frx.com
ORGANIZATION - Forest Research Institute
EDUCATION - Graduate College
LOCATION - Commack, NY, USA
SUBJECT_OF_QUESTION - Sputter Coater Recommendations
QUESTION - Hi folks,
I'm in need of recommendations (including both for and against)
regarding sputter coaters and their vendors. The application is SEM of
pharmaceutical materials. We have one now but it seems to be a complete
and total lemon as it lasts on average 3 weeks between needing
repairs... Vendors are welcome to contact me off list.

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Hi Robyn,

Glow discharge is the generation of plasma by an electric current. On a
coater or other machine that uses a glow discharge or sputter process,
polarity refers to which way the head is biased (positive or negative)
relative to the sample (ground). When the plasma is generated, positive
ions will be accelerated towards the negative end of the circuit and
vice versa. So polarity controls whether your sample is bombarded by
cations or anions. Current is the number of electrons flowing through
the generated plasma to complete the circuit and can be considered
proportional to the amount of plasma generated.

In sputtering, the plasma is generated from a noble gas with the
intention of bombarding a target with a process gas to free material
from the target and coat a sample. In glow discharge for grid
preparation, the plasma is made up from atmosphere (Oxygen, Nitrogen)
which is very reactive. The end goal is to make the grid hydrophilic,
which would mean adding more polar character (breaking C-C/C=C and C-H
bonds to make various bonds with Nitrogen and Oxygen).

-Jacob

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} Title-Subject: [Filtered] Glow Discharge
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First sentence is problematic. Instead, glow discharge occurs when the
generation of plasma is sustained in a chamber at right conditions of V
(electric field), P, and species. Current is a result of glow discharge.

The rest is fine.

On 8/28/13 12:06 PM, "jkabel-at-mail.ubc.ca" {jkabel-at-mail.ubc.ca} wrote:

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Position Opening at:

RESEARCH RESOURCES CENTER,
UNIVERSITY OF ILLINOIS AT CHICAGO

Visiting Research Specialist- Microscopy Specialist

The Electron Microscopy Service (EMS) of the Research Resources Center
(RRC) at the University of Illinois at Chicago has an open position for a
Visiting Research Specialist- Microscopy Specialist. The facility provides
electron and laser microscopy and surface analysis services for the
university research community and external organizations from two sites on
the campus. The open position is in the EMS-West facility, which
specializes in life science TEM and life and materials science SEM. The
west side facility includes a JEOL JEM-1220 TEM, Hitachi S-3000N VPSEM and
a JEOL JSM-6320F FESEM.

Qualified candidates must have a bachelor�s degree (preferably a master�s
degree or higher) in a related field, with at least three years electron
microscopy experience in SEM. They will help supervise the operation of
the specimen preparation area, including record keeping and maintenance
and will assist/supervise in the day to day running of the Electron
Microscopes. Interpersonal/ Communications skills are important, as this
individual will work with users, provide technical advice and demonstrate
how microscopy can advance their research.

Further information may be obtained by consulting the following website:
http://www.rrc.uic.edu/

Applications must be submitted through the hire touch system at
https://jobs.uic.edu/default.cfm?page=job&jobID=33863 . For fullest
consideration, interested parties should send an application no later than
September 22, 2013 with a cover letter, complete curriculum vitae, and the
names and addresses of three references. Please address all materials to
Alan W Nicholls, Ph.D.

UIC is an EEO/AA.

--
Alan W Nicholls, PhD
Associate Director, RRC
Director Research Resource Facility - Electron Microscopy
Research Resources Center, University of Illinois at Chicago
Rm 110, 845 West Taylor St
Chicago, IL 60607
312 996 1227
nicholls-at-uic.edu

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Hi

There are many problems that would give that which you outline but I believe
this instrument has a gas filled high voltage tank so please check the
pressure indicated on the tank. The operation manual should give you the
correct pressure reading an how to top up the tank.

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

X-from: zhangyc-at-gmail.com ()

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Email: zhangyc-at-gmail.com
Name: Yucheng Zhang

Title-Subject: [Filtered] TEM HT problem

Message: We have recently acquired a second-handed JEOL-2010 TEM microscope.
Unfortunately, some prob-lems seem to occur to the high tension. The current
emission quickly goes up to an abnormally large value when the acceleration
voltage is slowly increased to a certain value (e.g. 50 kV). The HT tank
also makes noise (sounds likes discharge). We have tried to manually
increase the voltage extremly slow (1kV/hour) to check if it will stabilize,
but it fails to work. Any advice on the problem would be much appreciated.
Thanks in advance.

Yucheng

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Hi all,

One of my users is having problems with antibody penetration into
frozen sections of brain from perfused (4% PFA) mice. The mice are
GFP+ and the GFP signal plus DAPI staining are fairly consistent
through the depth of the section, so we know the optics are fine.
However signal from two different primary and secondary antibodies
(AF555 and 633) exists only in the top few um of a 25-um thick section
after 2 days incubating at room temp.; one antibody seems to penetrate
a bit deeper than the other.

Someone suggested we try 20% DMSO in the blocking buffer and in the
antibody solutions, as she typically does that for whole-mount
staining for better penetration.

Any suggestions on what the problem could be and/or how to
troubleshoot are much appreciated.

Thanks!

-Esteban

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6, 29 -- Subject: LM (confocal): antibody penetration in frozen sections
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Dear Esteban,

usually penetration is more restricted for the labelled molecule (secondary antibody, protein A, Streptavidin etc) than for the primary antibody since the labeled complexes are larger in size.

In general, you want to look at such issues from two sides: from the perspective of the section and from that of the labelling molecules.

- try to use antibody fragments rather than intact Ig, certainly for the label step. Whatever you may loose in binding stability is likely compensated for by more molecules penetrating and finding their targets.

- use the smallest labels. Good penetration and good ultrastructure are achievable with ultra small gold particles, not so much with 10nm and larger.

- NaBH4 treatment (1% in 0.1M Phosphate buffer, freshly prepared) in our hands helps ultra small gold Ig conjugates to penetrate in 0.5% GA fixed cultured cells. This might help also with your PF fixed material.

Harsher treatments may be more successful but will progressively have an increasingly negative influence on structural integrity, however, sometimes one has no choice. Such treatments usually include taking up detergents in one or more steps of the the fixation procedure. Most gentle is Saponin perhaps which needs to be included during the immuno incubation steps as well. Triton-X-100 can be used as a 0.1% solution in buffer after the aldehyde fixation. When all else fails, detergent can be added to the fixative, but this results in general in serious extraction.

The results need of course always be interpreted against the artefacts introduced by the applied procedures.

Let me know if I can be of more help, good luck

Jan leunissen
Aurion

i: www.aurion.nl

On 29/08/2013, at 11:03 AM, g.esteban.fernandez-at-gmail.com wrote:

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} Hi all,
}
} One of my users is having problems with antibody penetration into
} frozen sections of brain from perfused (4% PFA) mice. The mice are
} GFP+ and the GFP signal plus DAPI staining are fairly consistent
} through the depth of the section, so we know the optics are fine.
} However signal from two different primary and secondary antibodies
} (AF555 and 633) exists only in the top few um of a 25-um thick section
} after 2 days incubating at room temp.; one antibody seems to penetrate
} a bit deeper than the other.
}
} Someone suggested we try 20% DMSO in the blocking buffer and in the
} antibody solutions, as she typically does that for whole-mount
} staining for better penetration.
}
} Any suggestions on what the problem could be and/or how to
} troubleshoot are much appreciated.
}
} Thanks!
}
} -Esteban

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15, 41 -- Subject: Re: [Microscopy] LM (confocal): antibody penetration in frozen sections
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Hi, Esteban:

I have the following questions:

1. Was the brain tissue post-fixed by immersion after perfusion? If yes,
for how long?
2. Was the brain tissue cryo protected before freezing.
3. Were the frozen sections cryostat sections?
4. Were sections treated with permeabilizing reagent before labeling
5. If these were cryostat sections, were they placed on glass slides for
labeling or free-floating in solution for labeling?
6. What were the secondary conjugates used? Were they fluorescent dye
conjugated antibodies or the fluoronanogold type of conjugates? Whole IgG
or antibody fragments?
7. What proteins the primary antibodies were against? I assume AF555 and
633 are commercial antibodies. Do you have data sheets for these
antibodies?

Penetration can be affected by a number of items. More details would be
helpful for diagnosing the cause. Thanks.

Hong

On 8/28/13 7:06 PM, "g.esteban.fernandez-at-gmail.com"
{g.esteban.fernandez-at-gmail.com} wrote:

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Hi

In my knowledge, glow discharge is obtained with an AC 50/60 Hz voltage
and not a DC. With an AC voltage, all the surfaces are bombarded by the
ionised gaz, and if the current is high enough, there is together
sputtering and redeposition everywhere.

It's the cheapest plasma cleaner and can be home made very easy. One
need an autotransformer, a HV 1500 V transformer, an HV vacuum
feedtrough and un piece of SS wire as electrode. OK, for security, all
that in a box with a fuse and an On/Off switch is not bad...
Works well for week organic contaminations, but does not replace a RF
plasma cleaner with Ar/O2 or H2/02 mixture.

Hope it helps

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Mat�riaux de Strasbourg
D�partement Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail : Jacques.Faerber-at-ipcms.unistra.fr

Le 28/08/2013 18:15, jkabel-at-mail.ubc.ca a �crit :
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} Hi Robyn,
}
} Glow discharge is the generation of plasma by an electric current. On a
} coater or other machine that uses a glow discharge or sputter process,
} polarity refers to which way the head is biased (positive or negative)
} relative to the sample (ground). When the plasma is generated, positive
} ions will be accelerated towards the negative end of the circuit and
} vice versa. So polarity controls whether your sample is bombarded by
} cations or anions. Current is the number of electrons flowing through
} the generated plasma to complete the circuit and can be considered
} proportional to the amount of plasma generated.
}
} In sputtering, the plasma is generated from a noble gas with the
} intention of bombarding a target with a process gas to free material
} from the target and coat a sample. In glow discharge for grid
} preparation, the plasma is made up from atmosphere (Oxygen, Nitrogen)
} which is very reactive. The end goal is to make the grid hydrophilic,
} which would mean adding more polar character (breaking C-C/C=C and C-H
} bonds to make various bonds with Nitrogen and Oxygen).
}
} -Jacob
}
}
} On 13-08-27 06:34 PM, microscopylistserver-noreply-at-microscopy.com wrote:
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} } Email: robyn.leidel-at-utsouthwestern.edu
} } Name: Robyn
} }
} } Organization: UTSW
} }
} } Title-Subject: [Filtered] Glow Discharge
} }
} } Message: Is there any one that can SIMPLY explain theoretically what glow discharge is, how it works
} } and the relationship between current and polarity? Please?...
} }
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Oups !
I forgot one point : the list of component is what one need to add a
glow dicharge device to an existing vacuum chamber. Of coarse, one need
that vacuum chamber, a bell jar with its pumps or an evaporator. It does
not work at normal pressure ;-). It's a pity !

J. Faerber
IPCMS-DSI
Institut de Physique et Chimie des Mat�riaux de Strasbourg
D�partement Surfaces et Interfaces
23, rue de Loess ; BP43
67034 Strasbourg CEDEX 2
France

Tel 00 33(0)3 88 10 71 01
Fax 00 33(0)3 88 10 72 48
E-mail : Jacques.Faerber-at-ipcms.unistra.fr

Le 29/08/2013 09:49, jacques.faerber-at-ipcms.u-strasbg.fr a �crit :
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
}
} Hi
}
} In my knowledge, glow discharge is obtained with an AC 50/60 Hz voltage
} and not a DC. With an AC voltage, all the surfaces are bombarded by the
} ionised gaz, and if the current is high enough, there is together
} sputtering and redeposition everywhere.
}
} It's the cheapest plasma cleaner and can be home made very easy. One
} need an autotransformer, a HV 1500 V transformer, an HV vacuum
} feedtrough and un piece of SS wire as electrode. OK, for security, all
} that in a box with a fuse and an On/Off switch is not bad...
} Works well for week organic contaminations, but does not replace a RF
} plasma cleaner with Ar/O2 or H2/02 mixture.
}
} Hope it helps
}
} J. Faerber
} IPCMS-DSI
} Institut de Physique et Chimie des Mat�riaux de Strasbourg
} D�partement Surfaces et Interfaces
} 23, rue de Loess ; BP43
} 67034 Strasbourg CEDEX 2
} France
}
} Tel 00 33(0)3 88 10 71 01
} Fax 00 33(0)3 88 10 72 48
} E-mail : Jacques.Faerber-at-ipcms.unistra.fr
}
} Le 28/08/2013 18:15, jkabel-at-mail.ubc.ca a �crit :
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Hi Robyn,
} }
} } Glow discharge is the generation of plasma by an electric current. On a
} } coater or other machine that uses a glow discharge or sputter process,
} } polarity refers to which way the head is biased (positive or negative)
} } relative to the sample (ground). When the plasma is generated, positive
} } ions will be accelerated towards the negative end of the circuit and
} } vice versa. So polarity controls whether your sample is bombarded by
} } cations or anions. Current is the number of electrons flowing through
} } the generated plasma to complete the circuit and can be considered
} } proportional to the amount of plasma generated.
} }
} } In sputtering, the plasma is generated from a noble gas with the
} } intention of bombarding a target with a process gas to free material
} } from the target and coat a sample. In glow discharge for grid
} } preparation, the plasma is made up from atmosphere (Oxygen, Nitrogen)
} } which is very reactive. The end goal is to make the grid hydrophilic,
} } which would mean adding more polar character (breaking C-C/C=C and C-H
} } bonds to make various bonds with Nitrogen and Oxygen).
} }
} } -Jacob
} }
} }
} } On 13-08-27 06:34 PM, microscopylistserver-noreply-at-microscopy.com wrote:
} } } ----------------------------------------------------------------------------
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} } }
} } } This Question/Comment was submitted to the Microscopy Listserver
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} } } Email: robyn.leidel-at-utsouthwestern.edu
} } } Name: Robyn
} } }
} } } Organization: UTSW
} } }
} } } Title-Subject: [Filtered] Glow Discharge
} } }
} } } Message: Is there any one that can SIMPLY explain theoretically what glow discharge is, how it works
} } } and the relationship between current and polarity? Please?...
} } }
} } } Login Host: 129.112.109.43
} } } Listserver Email Form V - 20120416
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Email: andel-at-cperi.certh.gr
Name: Andreas Delimitis

Organization: CPERI/CERTH

Title-Subject: [Filtered] TEM shut down during Bake Out

Message: Dear List Members,

I am seeking advice as to what precautions have to be taken to switch back on a JEOL 2010 HRTEM
after a serious power cut. The TEM was under a Bake Out process, with all lenses having been
significantly heated up and with no cooling water flow in them.

Thanking you in advance,
Andreas.
-------------------------------------------------
Dr. Andreas Delimitis
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CPERI / CERTH
6th km. Charilaou - Thermis road
57001 Thermi, Thessaloniki
Greece
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Hi
Are your incubation times for the secondaries 2 days, or 2 days for the primaries and only 1-2 hours for the secondaries? Best approach is incubate secondaries for the same length of time as the primaries.

Regards,
Glen
Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Cellular Morphology Core
Center on Human Development and Disability
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
glenmac-at-uw.edu

On Aug 28, 2013, at 4:05 PM, g.esteban.fernandez-at-gmail.com wrote:

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} Hi all,
}
} One of my users is having problems with antibody penetration into
} frozen sections of brain from perfused (4% PFA) mice. The mice are
} GFP+ and the GFP signal plus DAPI staining are fairly consistent
} through the depth of the section, so we know the optics are fine.
} However signal from two different primary and secondary antibodies
} (AF555 and 633) exists only in the top few um of a 25-um thick section
} after 2 days incubating at room temp.; one antibody seems to penetrate
} a bit deeper than the other.
}
} Someone suggested we try 20% DMSO in the blocking buffer and in the
} antibody solutions, as she typically does that for whole-mount
} staining for better penetration.
}
} Any suggestions on what the problem could be and/or how to
} troubleshoot are much appreciated.
}
} Thanks!
}
} -Esteban
}
} ==============================Original Headers==============================
} 6, 29 -- From g.esteban.fernandez-at-gmail.com Wed Aug 28 18:03:41 2013
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} 6, 29 -- Subject: LM (confocal): antibody penetration in frozen sections
} 6, 29 -- From: "G. Esteban Fernandez" {g.esteban.fernandez-at-gmail.com}
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By now everyone knows I'm a Wired website junkie. If you follow the link you find some interesting photomicrographs on the microbiology of food. Yes, there are a lot of non-scientific photos, but the opening SEM image of Penicillium roqueforti is worth the click.

http://www.wired.com/wiredscience/2013/08/microbes-food-beer/?viewall=true

Stay safe..............
Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

==============================Original Headers==============================
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Email: amit.welcomes.u-at-gmail.com
Name: Amit Gupta

Organization: IISc

Title-Subject: [Filtered] STEM probe changing shape?

Message: Good Morning/evening everyone,
I am working on a JEOL 2100F recently while working on STEM i noticed that at very low mag image
observed tends to change to oval shape, then after some time it will become circular then again oval
in perpendicular direction, see link for images:

https://sites.google.com/site/auxilliarylinks/

Why is this happening? will it affect resolution? I want to do high resolution HAADF but can not
attain lattice fringes.

Also what is the best way to align STEM mode for highest possible resolution in HAADF? any thing i
should specifically keep in mind?
Thank you
Amit Gupta

PS-you can reply to microscopy list server only, i have to submit through online form as all my
mails are filtered due to attachments

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Dear Amit,

This is not due to probe shape changes. If it were, the resolution of
your image would change across the field of view. Indeed, this happens
at very low magnifications (you can see the stigmatism in the
Ronchigram, especially towards the corners of the images where the
displacement from the optic axis is large), but at such low
magnifications you cannot detect the loss of the resolution.

I suspect you are seeing the differential pumping aperture (DPA) that
sits above the viewing chamber, just above the HAADF detector. I say
this because the edge of the shadow is not sharp.

The diffraction pattern you see while in STEM is formed by a real-space
image of the probe and sample in the DPA. If the microscope is properly
aligned, you should not see the DPA unless you work at low
magnifications where the spherical aberration of the objective lens
displaces the probe/sample image significantly. However, the elliptical
nature of the shadow suggest several possibilities:

1. The STEM scan pivot points are not set up properly (does the probe
move when tilting in imaging mode)? Indeed one, the excitation of the
deflectors is not symmetrical.

2. You have a very highly excited stigmator (probably objective or
diffraction stigmator).

3. The projector lenses are not at their correct (default) settings,
i.e. the crossover in the DPA is slightly above/below the correct plane.

Unless you're very familiar with the STEM alignments on a JEOL, I would
suggest you have JEOL take a look at the STEM alignments.

I hope that helps.

Best, Jon

} Title-Subject: [Filtered] STEM probe changing shape?
}
} Message: Good Morning/evening everyone,
} I am working on a JEOL 2100F recently while working on STEM i noticed
} that at very low mag image
} observed tends to change to oval shape, then after some time it will
} become circular then again oval
} in perpendicular direction, see link for images:
}
} https://sites.google.com/site/auxilliarylinks/
}
} Why is this happening? will it affect resolution? I want to do high
} resolution HAADF but can not
} attain lattice fringes.
}
} Also what is the best way to align STEM mode for highest possible
} resolution in HAADF? any thing i
} should specifically keep in mind?
} Thank you
} Amit Gupta

==============================Original Headers==============================
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Hi Amit,

the two images you show are taken at different magnifications and
neither is actually very low.

The circular shadow seen in Bright Field surrounding the image could be
the limit of the illumination, try different condenser aperture sizes to
see if that varies the diameter of the shadow, it's not important that
you can see it but it shouldn't be oval shaped and shouldn't change
direction, that's not right. What is intriguing is that the diameter of
the shadow that I have assumed is the condenser aperture limit does not
change with magnification in your two images (one image is 25K and the
other is 60K)

So the other thing to try is the camera length and perhaps the
intermediate stigmator has a fault on it that can affect the shape of
the illumination reaching the BF detector. The images look fine so the
condenser and objective are doing their jobs alright and the beam on the
sample is nice and round. Can you read the HEX values of the deflectors
when the "fault" appears and see what changes?

What kind of lattice images are you looking for and do you have an
aberration corrector? Alignment can be as easy as using the HT wobbler
and using the PL Align to shift the zoom point to the center of the
detectors. What probe size are you using, how strong is your C1 lens;
what illumination angle are you using, is it the optimum one? I'd get
an engineer to come and check things out.

Good luck
Rob

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} Title-Subject: [Filtered] STEM probe changing shape?
}
} Message: Good Morning/evening everyone,
} I am working on a JEOL 2100F recently while working on STEM i noticed that at very low mag image
} observed tends to change to oval shape, then after some time it will become circular then again oval
} in perpendicular direction, see link for images:
}
} https://sites.google.com/site/auxilliarylinks/
}
} Why is this happening? will it affect resolution? I want to do high resolution HAADF but can not
} attain lattice fringes.
}
} Also what is the best way to align STEM mode for highest possible resolution in HAADF? any thing i
} should specifically keep in mind?
} Thank you
} Amit Gupta
}
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--
Robert Keyse, DPhil
Research Scientist
Lehigh University

5, East Packer Avenue,
Whitaker Laboratory
Bethlehem,
PA 18015-3194

Office (610)758-3465
Fax (610)758-4244

==============================Original Headers==============================
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Hello Amit,
The images you showed are at different magnifications.

The shape change of your STEM image you mentioned

} "... image observed tends
} to change to oval shape, then after some time it will become circular
} then again oval in perpendicular direction"

depends on magnification change or occurred at specific magnification?

If the shape of your image is changing with magnification change this
might be caused by bad alignment of STEM.

I do not have any experiences with JEOL STEM systems, but in Philips
CM12/STEM such behavior of STEM image was caused by bad eucentric
position adjustment in Nanoprobe (STEM) alignment procedure.

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology, AS CR
Videnska 1083
CZ-142 20 Prague 4
Czech Republic

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} Title-Subject: [Filtered] STEM probe changing shape?
}
} Message: Good Morning/evening everyone,
} I am working on a JEOL 2100F recently while working on STEM i noticed
} that at very low mag image observed tends to change to oval shape,
} then after some time it will become circular then again oval in
} perpendicular direction, see link for images:
}
} https://sites.google.com/site/auxilliarylinks/
}
} Why is this happening? will it affect resolution? I want to do high
} resolution HAADF but can not attain lattice fringes.
}
} Also what is the best way to align STEM mode for highest possible
} resolution in HAADF? any thing i should specifically keep in mind?
} Thank you
} Amit Gupta
}
} PS-you can reply to microscopy list server only, i have to submit
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11, 41 -- From benada-at-biomed.cas.cz Fri Aug 30 08:53:41 2013
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Are you sure the primaries are all IgG's? I suspect you aren't using monoclonals since this is mouse tissue but a small percentage of monoclonals are IgM. And "poorly" made polyclonal sera can have a significant IgM component if they include antibodies from early bleeds. I also think anti-carbohydrate antibodies tend to be more likely IgM for some reason. IgM is, of course, 5x bigger than IgG. I have had this problem in my own lab with some homemade monoclonals. If the problem is from the secondaries, it should be easy to tell by switching the labels on the two different secondaries and seeing if the currently deep penetrating primary antibody is still detected.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: g.esteban.fernandez-at-gmail.com [mailto:g.esteban.fernandez-at-gmail.com]
Sent: Wednesday, August 28, 2013 6:05 PM
To: Phillips, Thomas E.

Hi all,

One of my users is having problems with antibody penetration into frozen sections of brain from perfused (4% PFA) mice. The mice are
GFP+ and the GFP signal plus DAPI staining are fairly consistent
through the depth of the section, so we know the optics are fine.
However signal from two different primary and secondary antibodies
(AF555 and 633) exists only in the top few um of a 25-um thick section after 2 days incubating at room temp.; one antibody seems to penetrate a bit deeper than the other.

Someone suggested we try 20% DMSO in the blocking buffer and in the antibody solutions, as she typically does that for whole-mount staining for better penetration.

Any suggestions on what the problem could be and/or how to troubleshoot are much appreciated.

Thanks!

-Esteban

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Thank you to everyone who provided some advice about TEM calibration grids.
It really helped to get me on the right track.

Summing up roughly

Catalase crystals :�� ~US$70, crystal lattice (8.75x6.85 nm).

Potassium chloroplatinate: ~US$40, planar spacing of 0.56nm

Copper phalocyanine: (1nm)

Crystal gold foil: suitable for very high magnifications, diffraction etc ~US$170 (0.2-0.1 nm)

Grahitised Carbon: ~US$40, suitable for high magnifications (plane spacing 0.34 nm)

Mag*I*Cal: ~US$1500, ion milled silicon/germanium grating. Unique in that it covers most of the magnification range of TEM (0.3 nm silicon lattice, ~10 nm line, ~1.2 �m between sets of lines, ~5.2 �m calibration area). I can see that this would be totally worth it for some people, just don't sneeze while holding that grid. I'll probably go for something cheaper first.

- Duane Harland

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Greetings fellow microscopists,

I am writing on behalf of a colleague at a nearby university who's draconian IT policy has cruelly banned this and other listservers.

He has just moved a Leo 440 SEM from one building to another and is trying to get it up and running.
Everything went well with one exception; the filament will not light. Good vacuum, HT etc.
The control system shows that only 1 of 2 ticks are lit for filament.

So two things.
1. Have any of you Leo users out there encountered this problem before?
2. Do you have any circuit diagrams that might be useful? (my friend has only a 1 page combined system diagram that does not show sufficient detail to be useful).

Kind regards,
Duane Harland
AgResearch, Lincoln, New Zealand

PS. Apologies for any mistakes in the description, I have not actually seen this machine myself, just discussed the situation by phone.

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The usual suspects have already been recommended: Williams & Carter for
TEM, Goldstein et al. for SEM/EDS, Bozzola for biological EM, Hibbs and
Price & Jerome for confocal light microscopy, and Sawyer and Grubb for
polymer microscopy. Any others?

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realname - Alejanro Hinojos
Email - ahinojos2-at-miners.utep.edu
ORGANIZATION - University of Texas at El Paso
EDUCATION - Undergraduate College
LOCATION - El Paso, TX, USA
SUBJECT_OF_QUESTION - Reference Books
QUESTION - Dear Microscopist,
My name is Alejandro Hinojos from the University of Texas at El Paso. I
am having difficulty picking out a book for microscopy. I have used
George F. Vander Voort's "Metallography: Principles and Practice" so far
and its text is good but not enough as in depth as I am looking for.
Thus I was wondering what you could suggest to me as a good reference
book for both electrical an optical microscopy.

==============================Original Headers==============================
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It's not a conventional textbook, but I have found the following book really useful especially because the same technique is sometimes called several different things depending on the science/engineering discipline. It has a good list of acronyms at the start too, which is great since many people at conferences etc assume that everyone knows what FLAP or SPLEEM are.
"Microscopy and Analysis Dictionary of Microscopy" by Julian P. Heath and published by Wiley

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Wednesday, 4 September 2013 9:07 a.m.
To: Harland, Duane

The usual suspects have already been recommended: Williams & Carter for TEM, Goldstein et al. for SEM/EDS, Bozzola for biological EM, Hibbs and Price & Jerome for confocal light microscopy, and Sawyer and Grubb for polymer microscopy. Any others?

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realname - Alejanro Hinojos
Email - ahinojos2-at-miners.utep.edu
ORGANIZATION - University of Texas at El Paso EDUCATION - Undergraduate College LOCATION - El Paso, TX, USA SUBJECT_OF_QUESTION - Reference Books QUESTION - Dear Microscopist, My name is Alejandro Hinojos from the University of Texas at El Paso. I am having difficulty picking out a book for microscopy. I have used George F. Vander Voort's "Metallography: Principles and Practice" so far and its text is good but not enough as in depth as I am looking for.
Thus I was wondering what you could suggest to me as a good reference book for both electrical an optical microscopy.

==============================Original Headers==============================
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Forgive me for answering this question, Esteban and Feng.

Both primary antibodies used were commercial antibodies against proteins
associated with synapse. One was a unpurified rabbit serum, and the other,
a purified mouse antibody isotype IgG2a.

As far as I know, mouse primary antibodies have been used often to bind
proteins in mouse tissue with clean labelings. I would like to know the
concern over this practice.

Thank you all.

Hong

On 9/1/13 5:18 PM, "PhillipsT-at-missouri.edu" {PhillipsT-at-missouri.edu} wrote:

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The first bleeds from a rabbit are often high in IgM's but presumably a decent quality commercial polyclonal would avoid this pitfall. I would be more cautious on a polyclonal I got from a colleague.

Mouse on mouse can certainly work but one needs to be cautious. Some tissues have cell types with Fc receptors that can bind the primary. Other tissues have endogenous mouse Ig's that bind the secondary. Trust but verify is always a good idea but especially important in using same species antibodies.

If I remember mouse IgG2a is more likely anti-protein epitopes than IgG1 (more common for carbohydrate antigens) and therefore less cross-reactivity of shared epitopes.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

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What are the other cell types that may also have Fc receptors? My
understanding is Fc receptors are mostly present on the surface of blood
cells. So unless I am labeling an antigen on blood cells, I should be
able to tell on EM my specific labeling, for example in synapse, from
potential background labeling due the Fc receptor binding on blood cell
surface, right? About the endogenous IgG, do they cross the brain-blood
barrier when animal is alive (the original posting was about labeling in
brain tissue)? If not, then they should be pretty much confined within
the blood vessels by perfusion fixation, right?

Another question. Tissues for immuno labeling are usually chemically
fixed before labeling. How effective the Fc receptor binding still is
after fixation?

Last one. How species specific the Fc receptor is? Would blocking with
normal serum (which contains IgG) before primary antibody incubation
saturate the Fc receptors in the tissue even if they are still functional
post fixation? If that is the case, would Fc receptors still be able to
bind the primary antibody?

Yes, a good control is important in the case of same species labeling.

Would love to hear more opinions. Thank you all and good night.

Hong

On 9/3/13 7:38 PM, "PhillipsT-at-missouri.edu" {PhillipsT-at-missouri.edu} wrote:

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Email: csown-at-voxa.co
Name: Chris Own

Organization: Voxa

Title-Subject: [Filtered] Seeking Gatan Firewire Adapter

Message: I received an e-mail asking for clarification of the Gatan equipment I'm seeking:

To clarify, I am looking to find a Gatan Firewire Adapter (GFA) Model 820. It is a little black box
that translates from the Digiscan D-sub37 connector to Firewire, enabling communication with the
computer.

Here is a photograph of the adapter needed:
https://dl.dropboxusercontent.com/u/4730981/Gatan%20GFA%20Model%20820.jpg

Help locating one for loan or purchase would be much appreciated!

Chris Own
Voxa

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Hello Everyone,
We just got an up-grade on our TEM camera and software. We replaced our 0.3meg Gatan etch-a-sketch with a new-to-us camera and Digital Micrograph 1.83.842 software.

I'm on my own for training and the help-me screen isn't very useful but I'm making progress. I'm currently stumped with spatial filters.

The instructions say "...select the image you want to (function)" and "Choose (function) and a new image will be created..." No matter what I do, I get a black image.

Any advice would be helpful!

Stay safe...........

Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Greetings

I teach bio EM in a community college certificate program.

I encourage my students to join the listserver and participate. I ask you to be kind and forgiving if anyone from my classes posts here. They are just beginning students, eager to join the microscopy community and sometimes need a little help getting started.

Students often come up with interesting questions. The latest one is 'What is it really like out in a job?' My job experience is limited, so I put it to all of you, what's it like out there. What would you tell a student about the world of work. They are hungry for anything, expectations of performance, chances for jobs, salaries, you name it, they want to know. So, if you are willing, send me things I can share with them and help them learn from all of us.

Thanks

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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I would like to add a phase contrast 5x objective to a Zeiss Axiovert 100. Zeiss says they no longer can supply that Ph0 phase ring, so I am planning to have my machine shop manufacture a metal disk of the correct size. If someone has one and can measure the diameter of the ring and the width of the opening, I would greatly appreciate it. Alternatively, if you know of an alternate source, I will explore that option. Thanks- Dave

David Knecht, Ph.D.
Professor and Head of Microscopy Facility
Department of Molecular and Cell Biology
U-3125
91 N. Eagleville Rd.
University of Connecticut
Storrs, CT 06269
860-486-2200
860-486-4331 (fax)

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Hi Frank,
just a basic suggestion, probably useless: did you check the Data Type
and the Contrast Limits of the new image? Some processing procedure ends
up with automatic contrast limits such that the processed image appears
black (at least with my DM version, which is quite older), especially if
the new image Data Type is Complex. This shouldn't be the case with
spatial filters, but never say never.
Hope this helps, though I'm not very confident

Davide

Il 04/09/2013 19.45, frank_karl-at-ardl.com ha scritto:
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} Hello Everyone,
} We just got an up-grade on our TEM camera and software. We replaced our 0.3meg Gatan etch-a-sketch with a new-to-us camera and Digital Micrograph 1.83.842 software.
}
} I'm on my own for training and the help-me screen isn't very useful but I'm making progress. I'm currently stumped with spatial filters.
}
} The instructions say "...select the image you want to (function)" and "Choose (function) and a new image will be created..." No matter what I do, I get a black image.
}
} Any advice would be helpful!
}
} Stay safe...........
}
} Frank
}
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} This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.
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Email: ravi.thakkar369-at-gmail.com
Name: Ravi

Title-Subject: [Filtered] Problem in Image AnalySIS S/W installed in FEI Tecnai 12.

Message: I am user of FEI Tecnai 12, we have Mega view II CCD camera with AnalySIS 3.2 (Build 678)
imaging software.
I have two queries :

1. How to get deleted images back from AnalySIS window Images deleted from AnalySIS S/W and not
saved in computer)

2. Image is very bright & less contrast if viewing frame is
passed through the bar of the TEM grid. If we move sample till bar go away from the viewing frame,
the same image having enough contrast.

Viewing frame without TEM grid bar
http://www.flickr.com/photos/97321550-at-N08/9676387161/

Viewing frame with TEM grid bar
http://www.flickr.com/photos/97321550-at-N08/9679620066/

But I have not seen this trouble with similar TEM & the
same AnalySIS S/W at other place.
So, Is there any setting to resolve this problem.

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Hello Ravi,
1 - the images are gone when you are closing the software without saving them first.
2- This is a problem of the region of interest (ROI) you selected for measuring the overall luminosity of the image. The "black
bar" and your "normal" image are spread too far concerning the grey values and the automatic tries to find a way to deal with them
all, resulting in a low contrast image. If your version of the software allows, try to use a "cross"-like measurement of the
intensities in the image (only using oneb horizontal and one vertical line in the image for measurement), or set the ROI smaller
and mid of the image. There should also be a "Camera Control" window where you can manipulate the greyvalues of the image before
aquisition.
Check also that in your Input channel or during post-processing you won`t cut too much of the shadow and highlight values of the
image (stay below 0,05%; I remember, default is ca. 0,1%).

If all this won`t help try to get in contact with Olympus Soft Imaging systems at Muenster, Germany. See www.olympus-sis.com

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 05.09.13 14:18, schrieb microscopylistserver-noreply-at-microscopy.com:
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} Title-Subject: [Filtered] Problem in Image AnalySIS S/W installed in FEI Tecnai 12.
}
} Message: I am user of FEI Tecnai 12, we have Mega view II CCD camera with AnalySIS 3.2 (Build 678)
} imaging software.
} I have two queries :
}
} 1. How to get deleted images back from AnalySIS window Images deleted from AnalySIS S/W and not
} saved in computer)
}
} 2. Image is very bright & less contrast if viewing frame is
} passed through the bar of the TEM grid. If we move sample till bar go away from the viewing frame,
} the same image having enough contrast.
}
}
} Viewing frame without TEM grid bar
} http://www.flickr.com/photos/97321550-at-N08/9676387161/
}
}
} Viewing frame with TEM grid bar
} http://www.flickr.com/photos/97321550-at-N08/9679620066/
}
}
}
} But I have not seen this trouble with similar TEM & the
} same AnalySIS S/W at other place.
} So, Is there any setting to resolve this problem.
}
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7, 23 -- From stefan.diller-at-t-online.de Thu Sep 5 08:40:49 2013
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I want to thank everyone for their replies. It turns out all I need to do was to click on the bottom bar of my Histogram icon and the filtered image contrast was reset and I got really nice images back.

Thanks again!!!!

-----Original Message-----
X-from: dcristofori-at-unive.it [mailto:dcristofori-at-unive.it]
Sent: Thursday, September 05, 2013 5:52 AM
To: Frank Karl

Hi Frank,
just a basic suggestion, probably useless: did you check the Data Type
and the Contrast Limits of the new image? Some processing procedure ends
up with automatic contrast limits such that the processed image appears
black (at least with my DM version, which is quite older), especially if
the new image Data Type is Complex. This shouldn't be the case with
spatial filters, but never say never.
Hope this helps, though I'm not very confident

Davide

Il 04/09/2013 19.45, frank_karl-at-ardl.com ha scritto:
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} Hello Everyone,
} We just got an up-grade on our TEM camera and software. We replaced our 0.3meg Gatan etch-a-sketch with a new-to-us camera and Digital Micrograph 1.83.842 software.
}
} I'm on my own for training and the help-me screen isn't very useful but I'm making progress. I'm currently stumped with spatial filters.
}
} The instructions say "...select the image you want to (function)" and "Choose (function) and a new image will be created..." No matter what I do, I get a black image.
}
} Any advice would be helpful!
}
} Stay safe...........
}
} Frank
}
}
}
} This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.
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This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Hello Ravi,

Stefan is correct on both counts.

Once you close the software without saving the images, they are deleted and
you can't recover them.
The issue with the black bar is due to the fact that the camera provides
images with more than 8 bits/pixel, and to display these on a monitor, the
contrast must be adjusted. When you move a sizable part of a grid bar into
the image, it will overwhelm the contrast of the sample, and you see the
effect that you describe. In addition to the solutions that Stefan proposed,
you can also set manual limits for the contrast and turn of the
"auto-contrast" function. This will avoid the contrast changes that you are
describing, but potentially leave you with sub optimum contrast in your live
images. Please note that the images are stored in 16-bit/pixel format
(unless you specifically set it to 8 bit), and will have the full
information, unless your images run into saturation. In other words, even if
the live image shows this contrast issue, the saved image will give you the
option to adjust the contrast manually and improve the contrast in the
sample area.

Best Regards,

Mike Bode, Ph.D.
ResAlta Research Technologies Corp.

2102 Beech Ct
Golden, CO 80401
USA

p: +1-303-234-9270
f: +1-303-202-6350
Mike.Bode at ResAltaTech.com
www.ResAltaTech.com

}
} X-from: ravi.thakkar369-at-gmail.com ()
}
} This Question/Comment was submitted to the Microscopy Listserver using
} the WWW based Form at
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Listserver
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} Email: ravi.thakkar369-at-gmail.com
} Name: Ravi
}
} Title-Subject: [Filtered] Problem in Image AnalySIS S/W installed in FEI
Tecnai 12.
}
} Message: I am user of FEI Tecnai 12, we have Mega view II CCD camera
} with AnalySIS 3.2 (Build 678) imaging software.
} I have two queries :
}
} 1. How to get deleted images back from AnalySIS window Images deleted
from AnalySIS S/W and not
} saved in computer)
}
} 2. Image is very bright & less contrast if viewing frame is
} passed through the bar of the TEM grid. If we move sample till bar go
away from the viewing frame,
} the same image having enough contrast.
}
}
} Viewing frame without TEM grid bar
} http://www.flickr.com/photos/97321550-at-N08/9676387161/
}
}
} Viewing frame with TEM grid bar
} http://www.flickr.com/photos/97321550-at-N08/9679620066/
}
}
}
} But I have not seen this trouble with similar TEM & the same AnalySIS
} S/W at other place.
} So, Is there any setting to resolve this problem.
}
} Login Host: 14.139.159.99
} Listserver Email Form V - 20120416
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==============================Original Headers==============================
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Since inquiring minds want to know.....
Our new-to-us camera is a Gatan Dual vision Mod780-p16FA. It was gifted to us when Firestone Tire up graded their STEM.

Stay safe........
Frank

-----Original Message-----
X-from: Monson, Frederick [mailto:FMonson-at-wcupa.edu]
Sent: Thursday, September 05, 2013 12:30 PM
To: Frank Karl

Hello Everyone,
We just got an up-grade on our TEM camera and software. We replaced our 0.3meg Gatan etch-a-sketch with a new-to-us camera and Digital Micrograph 1.83.842 software.

I'm on my own for training and the help-me screen isn't very useful but I'm making progress. I'm currently stumped with spatial filters.

The instructions say "...select the image you want to (function)" and "Choose (function) and a new image will be created..." No matter what I do, I get a black image.

Any advice would be helpful!

Stay safe...........

Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

==============================Original Headers==============================
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Name: Ram Chandra

Organization: Haldor Topsoe, Denmark

Title-Subject: [Filtered] Embedding Media for STEM/SEM EDS mapping

Message: Dear Fellow Microscopists,

I am recently looking into STEM EDS mapping of some ultramicrotomed samples. The problem is that we
have always used Epofix and Specifix for observing such samples in the TEM but they don't seem to be
too stable in the STEM.I have heard that acrylates are much better but have no experience with them.
Does anyone know resin that can be stable enough under EDS/WDS mapping conditions? Has anyone done
any mapping using aberration corrected STEM probe? Since we plan to do ultramicrotomy with the
samples I think i have to use a hard epoxy .. am I right? any help is welcome.

With best regards,
Ram

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Hi Ram,

Have you considered putting a thin carbon coat on your samples? I've
found that sections are much more resilient to the e-beam with a coarbon
coating. My experience is that uncoated epoxy sections "blow-up" the
moment I focus the e-beam on them. I assume that the carbon primarily
provides a conductive path reducing charge build-up. So whether the
sample blows apart to to electrostatic forces or decomposition of the
epoxy, the carbon makes a world of difference.

Cheers,
Henk

At 9/8/2013 7:03 PM, microscopylistserver-noreply-at-microscopy.com wrote:
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}
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}
} I am recently looking into STEM EDS mapping of some ultramicrotomed samples. The problem is that we
} have always used Epofix and Specifix for observing such samples in the TEM but they don't seem to be
} too stable in the STEM.I have heard that acrylates are much better but have no experience with them.
} Does anyone know resin that can be stable enough under EDS/WDS mapping conditions? Has anyone done
} any mapping using aberration corrected STEM probe? Since we plan to do ultramicrotomy with the
} samples I think i have to use a hard epoxy .. am I right? any help is welcome.
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} Ram
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--
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Hendrik O. Colijn
*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*
1305 Kinnear Rd, Suite 100, Columbus, OH 43212

614.643.3458 Office colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu}
www.cemas.osu.edu {http://www.cemas.osu.edu}

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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Hi all,

Can somebody advice a simple compact off-line program to view a large image resulted from montage of many smaller images ?
The key functionality required: easy zoom-unzoom, measure the distance between given points.

Thanks,
Pavel.

Pavel Potapov
GLOBALFOUNDRIES Dresden Module One LLC & Co. KG:

==============================Original Headers==============================
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Hi Pavel,
With ImageJ 1.47h and Windows 7 I can open an 0.5GB image with no problem
and make measurements. It is just necessary to set the memory buffer large
enough in the options for the program. I don't know the maximum file size
possible, but you can play with it.
Good luck.

Best Regards,
Larry Scipioni
--
Larry Scipioni, PhD
Vice President of R&D - Platform Development
ZS Genetics
les-at-zsgenetics.com
781-552-1989 (mobile)
larry.scipioni (Skype)
www.zsgenetics.com

-----Original Message-----
X-from: Pavel.Potapov-at-globalfoundries.com
[mailto:Pavel.Potapov-at-globalfoundries.com]
Sent: Monday, September 09, 2013 5:36 AM
To: LES-at-ZSGENETICS.COM

Hi all,

Can somebody advice a simple compact off-line program to view a large image
resulted from montage of many smaller images ?
The key functionality required: easy zoom-unzoom, measure the distance
between given points.

Thanks,
Pavel.

Pavel Potapov
GLOBALFOUNDRIES Dresden Module One LLC & Co. KG:

==============================Original Headers==============================
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Try grids with carbon film. Sections should behave nicely.
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

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Email: ramchandra.t-at-gmail.com
Name: Ram Chandra

Organization: Haldor Topsoe, Denmark

Title-Subject: [Filtered] Embedding Media for STEM/SEM EDS mapping

Message: Dear Fellow Microscopists,

I am recently looking into STEM EDS mapping of some ultramicrotomed samples. The problem is that we have always used Epofix and Specifix for observing such samples in the TEM but they don't seem to be too stable in the STEM.I have heard that acrylates are much better but have no experience with them.
Does anyone know resin that can be stable enough under EDS/WDS mapping conditions? Has anyone done any mapping using aberration corrected STEM probe? Since we plan to do ultramicrotomy with the samples I think i have to use a hard epoxy .. am I right? any help is welcome.

With best regards,
Ram

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Email: dwolfe-at-capitalhealth.org
Name: Dee Wolfe

Organization: Capital Health

Title-Subject: [Filtered] EM for Cadasil

Message: Good morning,

We have been asked to see if we can find a laboratory where we can submit a skin biopsy to test for
Cadasil (granular osmiophilic material, or GOM).

I am having no luck in this search. Would it be possible to see if any of your members offer this test?

Thanks in advance for any direction you can give us,

Dee Wolfe

Dee Wolfe MLT(ASCP), MT(HHS)
Anatomic Pathology Coordinator
Capital Health Regional Medical Center
Laboratory

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Email: lefevoura-at-mail.nih.gov
Name: Anthony LeFevour

Organization: NIH/NIDA/IRP/EM core

Title-Subject: [Filtered] EM core software for billing/scheduling/training

Message: Does anyone run a software like iLAB to facilitate scheduling/training/billing for an EM
core facility. I am tasked with setting up this structure at NIDA and could use your advise.

Thanks!

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I need to do some negative staining of antibodies. My uranyl formate is
long dead, and the uranyl acetate is a bit grainy. I have seen references
to but never tried sodium silicotungstate, although it seems to come
recommended for its fine grain size. I also see references to methylamine
tungstate, and I see EMS has sodium tungstate. I haven't used any of
these, only PTA, plus ammonium molybdate. Do any of you have any
recommendations for a fine-grained negative stain for e.g., antibodies and
proteins of similar size?

Mahalo from Honolulu,
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*
****************************************************************************

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Dear All
Since several weeks my Jeol JSM 5600 LV starts in a funny way: As I turn the
key the machine starts and proceeds immediately to evacuation and I can hear
the noise of the valves and see the evacuation light flushing. Evacuation
normally occurs about 15' after start. However, in about 5' a message
appears { {DP temperature low, pleae wait} } and evacuation stops, to start
again at about the normal time i.e. 10' later and then the microscope works
fine.
I wonder if anybody knows why this happens and if I have to worry about it.
Thanks
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

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Tina,

Please post a summary of responses that you get, or the off-line ones,
anyway. I'd like to get this information myself ...

Phil
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Hello Yorgos,
first check the clixxon temperature switch on the diff pump or its connecting cables to the pump logics board. There may be a
temporary shortcut somewhere with the effect that the pump logic sees a hot diffusion pump and starts the high vac cycle, where
there finally is a cold pump...
Check with Ohm-Meter; switch should be open when cold.

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 10.09.13 11:38, schrieb eikonika-at-otenet.gr:
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} ----------------------------------------------------------------------------
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} Dear All
} Since several weeks my Jeol JSM 5600 LV starts in a funny way: As I turn the
} key the machine starts and proceeds immediately to evacuation and I can hear
} the noise of the valves and see the evacuation light flushing. Evacuation
} normally occurs about 15' after start. However, in about 5' a message
} appears { {DP temperature low, pleae wait} } and evacuation stops, to start
} again at about the normal time i.e. 10' later and then the microscope works
} fine.
} I wonder if anybody knows why this happens and if I have to worry about it.
} Thanks
} yorgos
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} Tel/fax +30 210 8957677
} mobile +30 6945 107477
} www.eikonika.net www.aim.cat
} *************************************
}
}
} ==============================Original Headers==============================
} 4, 22 -- From eikonika-at-otenet.gr Tue Sep 10 04:33:38 2013
} 4, 22 -- Received: from sphinx.otenet.gr (smtp-out27.otenet.gr [83.235.69.27])
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} 4, 22 -- From: "yorgos nikas" {eikonika-at-otenet.gr}
} 4, 22 -- To: {microscopy-at-microscopy.com}
} 4, 22 -- Subject: Fw: Jeol JSM5600LV aberrant start-up procedure
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7, 22 -- From stefan.diller-at-t-online.de Tue Sep 10 09:12:20 2013
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Hello Stefan,

That's probably the origin of the problem. Nevertheless, for this type
of JEOL SEM, I believe the switch should be close when the DP is cold.
Best wishes,
Nicolas

Le 10/09/2013 16:18, stefan.diller-at-t-online.de a �crit :
} ----------------------------------------------------------------------------
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} ----------------------------------------------------------------------------
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} Hello Yorgos,
} first check the clixxon temperature switch on the diff pump or its connecting cables to the pump logics board. There may be a
} temporary shortcut somewhere with the effect that the pump logic sees a hot diffusion pump and starts the high vac cycle, where
} there finally is a cold pump...
} Check with Ohm-Meter; switch should be open when cold.
}
} Best wishes,
} Stefan
}
}
}
} -----------------------------------------------------
} Stefan Diller - Scientific Photography
} Arndtstrasse 22
} D - 97072 Wuerzburg Germany
} ++49-931-7848700 Phone
} ++49-931-7848701 Fax
} ++49-175-7177051 Mobile
}
} Websites:
} www.nanoflight.info
} www.stefan-diller.com
} www.electronmicroscopy.info
} www.elektronenmikroskopie.info
} www.zwillingsprojekt.de
} www.assisi.de
} Anfahrt: http://Mail.map24.com/Stefan.Diller
} -----------------------------------------------------
}
} Am 10.09.13 11:38, schrieb eikonika-at-otenet.gr:
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} } To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
} } On-Line Help http://www.microscopy.com/MicroscopyListserver/FAQ.html
} } ----------------------------------------------------------------------------
} }
} } Dear All
} } Since several weeks my Jeol JSM 5600 LV starts in a funny way: As I turn the
} } key the machine starts and proceeds immediately to evacuation and I can hear
} } the noise of the valves and see the evacuation light flushing. Evacuation
} } normally occurs about 15' after start. However, in about 5' a message
} } appears { {DP temperature low, pleae wait} } and evacuation stops, to start
} } again at about the normal time i.e. 10' later and then the microscope works
} } fine.
} } I wonder if anybody knows why this happens and if I have to worry about it.
} } Thanks
} } yorgos
} }
} } Dr Yorgos Nikas
} } Athens Innovative Microscopy
} } Skra 36 Voula 16673 GREECE
} }
} } Tel/fax +30 210 8957677
} } mobile +30 6945 107477
} } www.eikonika.net www.aim.cat
} } *************************************
} }
} }
} } ==============================Original Headers==============================
} } 4, 22 -- From eikonika-at-otenet.gr Tue Sep 10 04:33:38 2013
} } 4, 22 -- Received: from sphinx.otenet.gr (smtp-out27.otenet.gr [83.235.69.27])
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} 7, 22 -- From stefan.diller-at-t-online.de Tue Sep 10 09:12:20 2013
} 7, 22 -- Received: from mailout08.t-online.de (mailout08.t-online.de [194.25.134.20])
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--
Nicolas STEPHANT

Universit� de Nantes
Institut Jean Rouxel
Service de microscopie �lectronique � balayage et microanalyse
2 rue de la Houssini�re
BP 92208
44322 Nantes c�dex 3

T�l : 02 40 37 64 26
M�l : nicolas.stephant-at-univ-nantes.fr
Web : http://cnrs-imn.fr

"Le monde n'existe que pour autant que nous sommes capables d'en produire une image"
C.G Jung

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Dear Tina,
} I need to do some negative staining of antibodies.
this can be done but is tricky anyway - you won't see much, as far as I remember.
} My uranyl formate is long dead, and the uranyl acetate is a bit grainy.
sounds like carbonate precipitations / too high pH. You may try to prevent this by adding "a drop" of glacial acidic acid. Cave: I have never tried to rescue a "bad" batch of UAc solution, so I cannot comment whether this is waste of time.

} to but never tried sodium silicotungstate, although it seems to come
} recommended for its fine grain size. I also see references to methylamine
} tungstate, and I see EMS has sodium tungstate. I haven't used any of
} these, only PTA, plus ammonium molybdate.
or you also may try sodium (or potassium) phosphotungstate, 1%, at pH 6.5 to 7.0, which worked fine here in my lab for negative staining of both, viruses and proteins.
I think: All these solutions may work, and you will get recommendations from here and there.
The problem may come with the buffer in which the antibodies are dissolved - most likely PBS - and this may (!) result in precipitations. And the purity of the water matters, then the support film, and the kV of the TEM, and ...
Make one (or two, for comparison) of these solutions fresh. - Apply your antibody solution onto a carbon-coated 400 mesh /600 mesh copper grid (after hydrophilization / glow-discharge), wait for 30 to 120 sec (I am not patient, at all). Brief blotting, immediate wash for one second with bidist water, blot one second, then apply "your" staining solution, wait for 30 to 60 sec, and blot dry. - I am sure, you know this.
good luck and kind regards from Southern Germany,
Reinhard

--
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

Last microscopy conference:
http://www.mc2013.de/
MC2013 in Regensburg, Germany
Next microscopy conference:
http://www.imc2014.com/
18th IMC 2014 in Prague, Czech Rep.

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Dear List
Thanks to your comments I think I solved the problem. Usualy I open the
cooling water flow 10' after starting the machine. but today I opened it 10'
before starting.. I thought if the high vacuum cycle starts immediately
because the diffusion pump is perceived to be hot enough, coolling it in
advance with water may solve the problem. And indeed, the microscope started
normally!
Possibly the summer temperatures keeps the DP warm overnight untill next day
I start again.
I am very thankfull to Nicolas, Stefan, Rick and Phil for their help. This
List is a great tool!
Have a nice day
yorgos

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Title-Subject: [Filtered] Thanks!

Message: Thanks to all who replied to my post regarding cadasil. Who knew there would be so many
options to choose from after my web search had come up empty? I have forward your replies to our
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Hooke College of Applied Sciences, located in Westmont, IL, is offering a transmission electron microscopy short course October 1-3, 2013. In addition to lectures, this course emphasizes hands-on training using state-of-the-art equipment. For further TEM training details and registration information, please follow the link below:
http://www.hookecollege.com/courses/course.asp?COURSE_ID=53

October 21-25, 2013 a course in scanning electron microscopy will be offered at Hooke College of Applied Sciences. Further information about this hands-on course can be found at the link below:
http://www.hookecollege.com/courses/course.asp?COURSE_ID=42

Chris Gorman | 850 Pasquinelli Drive |Westmont, IL 60559 | 630-887-7100| 630-887-7412(f)

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I want to thank the people who replied. A few pointed out some new (to
me) stains available, including Blue Platinum from IBI Labs, NanoVan from
Nanoprobes, and a non-radioactive uranyl acetate replacement, UAR_EMS,
from EMS. Other stains I have not personally tried include sodium
silicotunsgtate and methylamine tungstate. Shipping radioactive or
hazardous chemicals to Hawaii is prohibitively expensive, so I'm looking
for an alternative to the really old, photodegraded uranyl formate I had
that had a fine grain size and high contrast.

Recommendations from people generally insisted that the uranyl stains need
to be fresh, and my stains were a bit old. In addition, I do not have any
pure carbon films right now. I used to make them by evaporating onto mica
and stripping them off, but my evaporator has a leak. I have to admit that
I have not repaired it because the leak is exactly the right amount for
glow discharging grids, so I have had little motivation to fix it. I have
not had much success with commercial carbon-with-removable-formvar grids.
You may detect some laziness on my part, so I will clean up my act and try
again!

To clarify what I am trying to do, a researcher has some evidence that
some antibodies from a sea creature that will for now be unnamed (not sure
how much I can disclose) have a strange configuration. In addition to the
typical "Y" shape, there is probably another smaller Y complex sticking
off to one side. I can see pretty well the larger protein loops, but I
need to be able to resolve some smaller structures. A smaller stain grain
size would help plus, obviously, cleaning up some background. My lazy
technique has been adequate for all these giant marine viruses we look at,
but I need to refine this to see smaller structures.

The sample is 1 ug/ml in sodium borate pH 8.0.

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*
****************************************************************************

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Hi Tina,
�
I am sure you have more than one argument to explain your summer laziness in a beautiful place like hawaii.
By reading your message about your loch-hawaiiness monster�I remembered an interesting instrument called low voltage electron microscope (LVEM).
Perhaps it is the right application for this instrument, because your main problem is the lack of contrast and the difficulty of developping a contrasting methodology.
Due to the low voltage of the LVEM, perhaps you can see your protein complex without contrasting at all.
I'll bet Delong would be interested to find a new application for their instrument if you care to contact them.
Otherwise I wanted to cite shadowing as an alternative to negative contrasting.
�
Good luck
Stephane

________________________________
X-from: "tina-at-pbrc.hawaii.edu" {tina-at-pbrc.hawaii.edu}
To: nizets2-at-yahoo.com
Sent: Wednesday, September 11, 2013 11:10 PM

I want to thank the people who replied.� A few pointed out some new (to
me) stains available, including Blue Platinum from IBI Labs, NanoVan from
Nanoprobes, and a non-radioactive uranyl acetate replacement, UAR_EMS,
from EMS.� Other stains I have not personally tried include sodium
silicotunsgtate and methylamine tungstate. Shipping radioactive or
hazardous chemicals to Hawaii is prohibitively expensive, so I'm looking
for an alternative to the really old, photodegraded uranyl formate I had
that had a fine grain size and high contrast.

Recommendations from people generally insisted that the uranyl stains need
to be fresh, and my stains were a bit old. In addition, I do not have any
pure carbon films right now. I used to make them by evaporating onto mica
and stripping them off, but my evaporator has a leak. I have to admit that
I have not repaired it because the leak is exactly the right amount for
glow discharging grids, so I have had little motivation to fix it. I have
not had much success with commercial carbon-with-removable-formvar grids.
You may detect some laziness on my part, so I will clean up my act and try
again!

To clarify what I am trying to do, a researcher has some evidence that
some antibodies from a sea creature that will for now be unnamed (not sure
how much I can disclose) have a strange configuration. In addition to the
typical "Y" shape, there is probably another smaller Y complex sticking
off to one side. I can see pretty well the larger protein loops, but I
need to be able to resolve some smaller structures. A smaller stain grain
size would help plus, obviously, cleaning up some background. My lazy
technique has been adequate for all these giant marine viruses we look at,
but I need to refine this to see smaller structures.

The sample is 1 ug/ml in sodium borate pH 8.0.

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*
****************************************************************************

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Name: Ram chandra

Organization: Haldor topsoe

Title-Subject: [Filtered] Re:Embedding Media for STEM/SEM EDS mapping- Thanks

Message:
Dear Friends,

Thank you very much for responding to my question. Based on the Input I have received, I think LR
white and Spurr equivalent would be the best choice. I will also coat them with carbon to see what
happens. The idea of
mixing nanotubes is also very interesting and out of the box but maybe complicated for the
beginning. (So, sorry Stephane.. may not develop a product and offer you royalties)
I do wonder if carbon coating a sample causes contamination issues in an aberration corrected
microscope.

Thanks a lot and Cheers,
Ram

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Two questions on immunocytochemistry using fluorescent probes that I should know the answer to but seem to have forgotten and can't find a definitive reference which answers it.

1. Does anybody know of a reference where they measured the average number of secondary antibodies that bind to a single primary? It is generally thought to be "several" but if the secondary is a polyclonal (say goat anti-mouse IgG(H+L), it might be reasonable to assume there would be at least 4 binding sites (one on each of the H chains, one on each of the L chains). But clearly one could have more. Has this been measured?

The optimal number of fluorochromes on an secondary is generally felt to be between 2 and 8 (some say 2 and 5). So if four secondary antibody molecules, each with 5-8 fluorophores attached, labeled of a single primary, there would be 20-32 fluorophores on it.

2. Ignoring super-resolution or TIRF microscopy approaches and using conventional widefield or confocal microscopy, within inherent background, quenching, etc., is there an estimate on the number of epitopes on a single organelle that need to be labeled to detect it? If you labeled one protein on a nuclear envelope with a single primary and 4 secondaries, would you see a single dot in widefield fluorescence microscopy? I realize in most cases you are labeling a ton of similar proteins on a target organelle but what is the lower limit that works? �

Thanks, Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

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Hi Ram,

Regarding contamination from the C coating. We generally don't see
any. The amorphous C coating is pretty tightly bonded.

Most of the contamination we see seems to be due to small organic
molecules which can rapidly diffuse across the sample under the
influence of the e-field from the beam. If you do see contamination, I
would suspect it comes from unreacted epoxy monomers.

Cheers,
Henk

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} white and Spurr equivalent would be the best choice. I will also coat them with carbon to see what
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} mixing nanotubes is also very interesting and out of the box but maybe complicated for the
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Hendrik O. Colijn
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1305 Kinnear Rd, Suite 100, Columbus, OH 43212

614.643.3458 Office colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu}
www.cemas.osu.edu {http://www.cemas.osu.edu}

"Time is that quality of nature which keeps things from happening all at
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I think I am doing a bad job of explaining my question so let me try again.

Super-resolution might not increase sensitivity but can involve doing things like subtracting images to detect bleaching or blink on events. Collection of sequential images using photo-activated probes is not really comparable to collecting a standard confocal or wide-field image of an immunostained tissue prep where you have saturated the tissue with 1-10 ug/ml of primary and secondary antibodies and have a background that you need to discern the signal from. A non-immunostaining analogy would be people looking at single RNA molecules who I seem to remember find it takes 4 - 32 binding sites for probes to allow the molecule to be seen against background (but perhaps I am remembering this wrong).

Lots of antigens we look for in tissue sections are highly abundant - a single cell might have several hundred thousand insulin receptors so succeeding at seeing that isn't amazing
When I do fluorescent lectin staining of sections, almost all probes give me some "labeling" but how many lectin molecules are bound to the apical membrane of a cell that is really dimly stained vs. one that knocks your socks off? When I have "low" labeling, I don't see a few random really bright dots but instead a dull even glow. I suspect that dull glow is still a lot of fluorophores and that if it had been a few molecules of lectin spatially restricted to a few nanometers of the apical membrane, I would never have recognized it.

I am not sure how to answer this question but perhaps if you had cells that had a small number of viral particles (literally less than 20 or something - I assume the virologists can determine that) and you had a monoclonal that could effectively label some epitope known to be present on the virus in some small number (once?), you could tell if test whether you could detect it. This has probably been done at the EM level using immunogold. Does anyone know a paper like this at the LM level using fluorescence?

Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: Stephane Nizet [mailto:nizets2-at-yahoo.com]
Sent: Friday, September 13, 2013 1:31 AM
To: Phillips, Thomas E.

Two questions on immunocytochemistry using fluorescent probes that I should know the answer to but seem to have forgotten and can't find a definitive reference which answers it.

1. Does anybody know of a reference where they measured the average number of secondary antibodies that bind to a single primary? It is generally thought to be "several" but if the secondary is a polyclonal (say goat anti-mouse IgG(H+L), it might be reasonable to assume there would be at least 4 binding sites (one on each of the H chains, one on each of the L chains). But clearly one could have more. Has this been measured?

The optimal number of fluorochromes on an secondary is generally felt to be between 2 and 8 (some say 2 and 5). So if four secondary antibody molecules, each with 5-8 fluorophores attached, labeled of a single primary, there would be 20-32 fluorophores on it.

2. Ignoring super-resolution or TIRF microscopy approaches and using conventional widefield or confocal microscopy, within inherent background, quenching, etc., is there an estimate on the number of epitopes on a single organelle that need to be labeled to detect it? If you labeled one protein on a nuclear envelope with a single primary and 4 secondaries, would you see a single dot in widefield fluorescence microscopy? I realize in most cases you are labeling a ton of similar proteins on a target organelle but what is the lower limit that works? �

Thanks, Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://www.biology.missouri.edu/faculty/phillips.html
http://www.biotech.missouri.edu/mcc/

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I certainly agree about the steric problem and that is why I asked if anyone has measured this.

The IgG does have two binding sites formed by the H + L chain but that is not what I used to make my assumption. I was proposing that an IgG has 4 chains - 2 x H and 2 x L. I was assuming each chain would have a minimum of 1 epitope recognized by the secondary. I realize some epitopes might be discontinuous targets that span an H + L chain. Mostly I was thinking the end of the Fc (end of the H) was pretty far from the L chain so there would be room for both. There should be more epitopes but that is where the steric issue comes up and why I asked if anyone has measure it. It would vary for the secondary but it is probably about the same for lots of secondary antibodies.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

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Our facility has a TPI Lancer Vibratome Series 1000. We're looking for �-inch and 1-inch specimen mounts for it. We're also in need of the serrated screw on the blade holder (the screw fastens the blade holder at the desired sectioning angle). I'm well aware that the model has changed hands and evolved but if anyone can help out, please let me know. (If anyone has this model and can give me the specifications for the screw threads, that would suffice as well.)

Thank you,

Jaci

Jaclynn Lett
Senior Research Technician, EM Facility
Research Center for Auditory and Vestibular Studies
Department of Otolaryngology
Washington University School of Medicine
660 S. Euclid Ave., Campus Box 8115
St. Louis, MO 63110
Office: 314-747-7257
Fax: 314-747-7230
Email: lettj-at-ent.wustl.edu
http://otocore.wustl.edu

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There is a discussion on the confocal listserver today that adds some insight to the question I raised by how many molecules of fluorophores we are seeing in a typical immunocytochemical staining.

Serkan Berk suggested a paper by Charpilienne et al. 2001 J. Biol. Chem 276, 29361-29367. (http://www.jbc.org/content/276/31/29361.full) titled "Individual Rotavirus-like Particles Containing 120 Molecules of Fluorescent Protein Are Visible in Living Cells" which adds some insight into my question.

Jolien Verdaasdonk in another posting suggested a paper from Ted Salmon's group Lawrimore et al. 2011 JCB vol. 195 no. 4 573-582 where they used a spinning disk confocal with Orca camera and clearly were showing point centromeres that they calculated contained 5 copies of GFP. GFP isn't as bright as an Alexa fluorochrome but on the other hand you don't have as big of problem of a background haze from non-specific binding of primary and secondaries.

I guess my problem is just integrating what I see when I read a beautiful paper like the Lawrimore one in which a few labels are sufficient to generate a punctate bright dot and then seeing low level membrane fluorescence which I have assumed was due to 10-100's of copies of lectin binding compared to } 10,000 copies binding when it was really bright.

Part of this came from my thinking about the number of secondary IgG's binding to a single primary and how many fluorophores that would be and then going off track. I must just be overthinking this... not the first time.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri(2001) JBC
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

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I have some users who have very few precious bacterrial samples. When they
tried enrobing them in agarose for fixation and resin embedding, not only
did the cells get a little too dispersed (I told them they need to
practice), but their tiny agar blocks were nearly invisible in the
unpolymerized and polymerized resin, and they lost some. I remember in the
dark past that the agarose could be stained with something that would
remain throught the processing and into the resin, possible something
blue. I can not find that reference now, and even with a very specific
wording, Google comes up with 8,080,000 results in 0.40 seconds.

Any favorite protocols gratefully entertained!

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*
****************************************************************************

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Tina,

Have you tried toludine blue? It's metachromatic for
mucopolysaccarhides. But as it's basic, it would have to be used after
fixation and any uranyl acetate en bloc stain. It should bind well
enough to the agarose to still be there when the resin steps are reached.
If the tol blue does want to come out of the agarose, a trick to try is
ammonium molybdate, 5% aqueous (must not be cloudy) for 5 min following
the tol blue, then rinse in "running tap water" (approximate this) for
2-3 minutes. The molybdate acts as a mordant, and after this treatment,
the tol blue won't come out in water or alcohol. Note: you may have to
use acidified tol blue to use molybdate. (Kiernan, "Histological and
Histochemical Methods: Theory and Practice" 3rd edition.)

Phil

} I have some users who have very few precious bacterrial samples. When they
} tried enrobing them in agarose for fixation and resin embedding, not only
} did the cells get a little too dispersed (I told them they need to
} practice), but their tiny agar blocks were nearly invisible in the
} unpolymerized and polymerized resin, and they lost some. I remember in the
} dark past that the agarose could be stained with something that would
} remain throught the processing and into the resin, possible something
} blue. I can not find that reference now, and even with a very specific
} wording, Google comes up with 8,080,000 results in 0.40 seconds.
}
} Any favorite protocols gratefully entertained!
}
} 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*
} ****************************************************************************
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Tina,
Back in the day, I did this with fast green. I made a
saturated solution in 1oo % ethanol, and then would add a microL per
sample vial (say 1 mL or so of ethanol) at the 100% stage. To be
fair, I was infiltrating in butyl methyl methacrylate not epoxy. It
is possible that the color would be more strongly extracted by those
resins. But the fast green did fine for the agarose in the
methacyrlate.

As ever
Tobias

} -
} ----------------------------------------------------------------------------
}
} I have some users who have very few precious bacterrial samples. When they
} tried enrobing them in agarose for fixation and resin embedding, not only
} did the cells get a little too dispersed (I told them they need to
} practice), but their tiny agar blocks were nearly invisible in the
} unpolymerized and polymerized resin, and they lost some. I remember in the
} dark past that the agarose could be stained with something that would
} remain throught the processing and into the resin, possible something
} blue. I can not find that reference now, and even with a very specific
} wording, Google comes up with 8,080,000 results in 0.40 seconds.
}
} Any favorite protocols gratefully entertained!
}
} 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*
} ****************************************************************************
}
} =

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Professor
/_ / __ /__ \ \ \__ Biology Department
/ / / \ \ \ 611 N. Pleasant St.
/ / / \ \ \ University of
Massachusetts
/ / ___ / \ \__/ \ ____ Amherst, MA, 01003
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
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4, 21 -- To: tina-at-pbrc.hawaii.edu
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Hi Tina:

We routinely use 2% potassium permanganate (KMnO4) to fix agarose embedded yeast cells, because osmium does not penetrate the yeast cell wall. One "by-product" of potassium permanganate fixation is that it completely turns the cells, and the agarose to black, making the block visible for sectioning.

I assume (never tried myself) osmium will do the same thing, except that the osmium won't penetrate the bacterial cell wall either. For the sake of "staining" the agarose, it might worth to try.

If you are doing immuno-gold labeling, however, that won't work! What we do is to try to identify the cells under a dissecting microscope, mark them with a sharpie and go from there.

Zhaojie Zhang, Ph.D.
Director, Microscopy Facility
University of Wyoming

-----Original Message-----
X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu]
Sent: Monday, September 16, 2013 8:17 AM
To: Z.J. Zhang

Tina,
Back in the day, I did this with fast green. I made a saturated solution in 1oo % ethanol, and then would add a microL per sample vial (say 1 mL or so of ethanol) at the 100% stage. To be fair, I was infiltrating in butyl methyl methacrylate not epoxy. It is possible that the color would be more strongly extracted by those resins. But the fast green did fine for the agarose in the methacyrlate.

As ever
Tobias

} -
} -----------------------------------------------------------------------
} -----
}
} I have some users who have very few precious bacterrial samples. When
} they tried enrobing them in agarose for fixation and resin embedding,
} not only did the cells get a little too dispersed (I told them they
} need to practice), but their tiny agar blocks were nearly invisible in
} the unpolymerized and polymerized resin, and they lost some. I remember
} in the dark past that the agarose could be stained with something that
} would remain throught the processing and into the resin, possible
} something blue. I can not find that reference now, and even with a very
} specific wording, Google comes up with 8,080,000 results in 0.40 seconds.
}
} Any favorite protocols gratefully entertained!
}
} 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*
} ***********************************************************************
} *****
}
} =

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Professor
/_ / __ /__ \ \ \__ Biology Department
/ / / \ \ \ 611 N. Pleasant St.
/ / / \ \ \ University of
Massachusetts
/ / ___ / \ \__/ \ ____ Amherst, MA, 01003
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
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4, 21 -- Date: Mon, 16 Sep 2013 10:13:38 -0400 4, 21 -- To: tina-at-pbrc.hawaii.edu 4, 21 -- From: Tobias Baskin {baskin-at-bio.umass.edu} 4, 21 -- Subject: Re: [Microscopy] Staining of agarose for TEM embedding 4, 21 -- Cc: microscopy-at-microscopy.com 4, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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17, 31 -- From ZZhang-at-uwyo.edu Mon Sep 16 10:47:01 2013
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17, 31 -- Subject: RE: [Microscopy] Re: Staining of agarose for TEM embedding
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When I send my blocks to a campus facility for paraffin processing and embedding, I need to mark one of the edges so they can know the orientation I want. They gave me a little vial of tattoo ink pigment in powder form. I usually add it to the connective tissue side opposite the epithelial surface I am interested in using a wooden applicator. The big pigment particles adhere pretty well and easy to see. I suspect you could add a couple of tiny particles from it to your agarose and be able to see it in a stereoscope. I haven't tried this for EM blocks but suspect it would work. This might minimize the chance a stain would interact with your bacteria. Tom

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
X-from: ZZhang-at-uwyo.edu [mailto:ZZhang-at-uwyo.edu]
Sent: Monday, September 16, 2013 10:48 AM
To: Phillips, Thomas E.

Hi Tina:

We routinely use 2% potassium permanganate (KMnO4) to fix agarose embedded yeast cells, because osmium does not penetrate the yeast cell wall. One "by-product" of potassium permanganate fixation is that it completely turns the cells, and the agarose to black, making the block visible for sectioning.

I assume (never tried myself) osmium will do the same thing, except that the osmium won't penetrate the bacterial cell wall either. For the sake of "staining" the agarose, it might worth to try.

If you are doing immuno-gold labeling, however, that won't work! What we do is to try to identify the cells under a dissecting microscope, mark them with a sharpie and go from there.

Zhaojie Zhang, Ph.D.
Director, Microscopy Facility
University of Wyoming

-----Original Message-----
X-from: baskin-at-bio.umass.edu [mailto:baskin-at-bio.umass.edu]
Sent: Monday, September 16, 2013 8:17 AM
To: Z.J. Zhang

Tina,
Back in the day, I did this with fast green. I made a saturated solution in 1oo % ethanol, and then would add a microL per sample vial (say 1 mL or so of ethanol) at the 100% stage. To be fair, I was infiltrating in butyl methyl methacrylate not epoxy. It is possible that the color would be more strongly extracted by those resins. But the fast green did fine for the agarose in the methacyrlate.

As ever
Tobias

} -
} -----------------------------------------------------------------------
} -----
}
} I have some users who have very few precious bacterrial samples. When
} they tried enrobing them in agarose for fixation and resin embedding,
} not only did the cells get a little too dispersed (I told them they
} need to practice), but their tiny agar blocks were nearly invisible in
} the unpolymerized and polymerized resin, and they lost some. I remember
} in the dark past that the agarose could be stained with something that
} would remain throught the processing and into the resin, possible
} something blue. I can not find that reference now, and even with a very
} specific wording, Google comes up with 8,080,000 results in 0.40 seconds.
}
} Any favorite protocols gratefully entertained!
}
} 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*
} ***********************************************************************
} *****
}
} =

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Professor
/_ / __ /__ \ \ \__ Biology Department
/ / / \ \ \ 611 N. Pleasant St.
/ / / \ \ \ University of
Massachusetts
/ / ___ / \ \__/ \ ____ Amherst, MA, 01003
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

==============================Original Headers==============================
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4, 21 -- Date: Mon, 16 Sep 2013 10:13:38 -0400 4, 21 -- To: tina-at-pbrc.hawaii.edu 4, 21 -- From: Tobias Baskin {baskin-at-bio.umass.edu} 4, 21 -- Subject: Re: [Microscopy] Staining of agarose for TEM embedding 4, 21 -- Cc: microscopy-at-microscopy.com 4, 21 -- Content-Type: text/plain; charset="us-ascii" ; format="flowed"
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Email: lciampa-at-uwyo.edu
Name: Lyn Ciampa

Organization: UWyo Plant Sciences

Title-Subject: [Filtered] Extranenous prepared slides

Message: Hi all,
I am working developing my compound microscopy skills and would like to obtain quality prepared
slides (something improved over Carolina Fischer scientific, etc. Preferably these would be
botanical (I am an agroecology major). I am wondering if anyone here has any to pass on,sell, etc. I
would send post paid mailers to make it less of a hassle.

Thanks,
Lyn

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Hi Lyn,

I think the best thing to do would be to make hand sections, mount them in
water with a coverslip, and look at these. You can look at them unstained
or stained with something like toluidine blue. Cheap, easy and you learn
much more. This is the way I've taught first- and second-year university
students and also high school students. Doesn't take long to get good
sections. If you're not sure about this, I can send a pdf on hand
sectioning.

cheers,
Rosemary

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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Dear Fellow Microscopists,

We have to replace a HT cable in our 20 years old Philips CM-20
SuperTwin TEM. Since a brand new cable is very expensive (what hurts
even more because of the age of the microscope) we are looking for an
alternative. If you know any (spare parts, microscope to be scrapped,
etc) please let me know.

With best regards,
Leszek

Leszek Kepinski
Institute of Low Temperature and Structure Research,
Polish Academy of Sciences,
Okolna 2, 50-422 Wrocław, Poland
e-mail: l.kepinski-at-int.pan.wroc.pl

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Hi Leszek

There are many organisation that work with high voltage systems and I have
many times, in a number of countries within Europe, taken the damaged cable
to such an organisation and had a new cable built using the original cable
ends.

Organisations who deal with medical equipment, x-ray sets etc are able to
help in these cases.

Good luck

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: L.Kepinski-at-int.pan.wroc.pl [mailto:L.Kepinski-at-int.pan.wroc.pl]
Sent: 17 September 2013 14:01
To: protrain-at-emcourses.com

Dear Fellow Microscopists,

We have to replace a HT cable in our 20 years old Philips CM-20 SuperTwin
TEM. Since a brand new cable is very expensive (what hurts even more because
of the age of the microscope) we are looking for an alternative. If you know
any (spare parts, microscope to be scrapped,
etc) please let me know.

With best regards,
Leszek

Leszek Kepinski
Institute of Low Temperature and Structure Research, Polish Academy of
Sciences, Okolna 2, 50-422 Wrocław, Poland
e-mail: l.kepinski-at-int.pan.wroc.pl

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Greetings

So I was sitting at my desk waiting to start my digital imaging class and got to thinking, 'What should really be in this class?'

I wonder about things like this a lot and usually end up going with the established topics and tell myself I should really look into this sometime.

Well, the time is now. Who else to query about this than the very people who do this kind of thing everyday and who might be the future employers of my students?

Here's the deal. We are a community college that offers certificates in electron microscopy, both biological and materials techniques. Most of our students have the intention of getting a technician level job, some may go on to a 4 year degree, but not all. What do you think they need to know about digital imaging to be successful?

The class now covers the hardware, CCD's, CMOS, image displays, printing technologies, basic image capture software (the ones we have on our scopes), Gatan DM, ImageJ, basic Photoshop, Illustrator, PowerPoint etc. We talk about imaging ethics and read instructions to authors for guidance. I do not cover things like Photoshop in much detail, thinking that most of Photoshop is off limits to us.

My background is mostly self taught. I was a whiz in the darkroom for many years, so I picked up digital imaging as we converted from film. I had a lot of the basic background info about what makes a good picture under my belt before I got started. Lots of my students do not have this background.

We used to offer this class as a 'dessert' after the students had done everything else, but now I am wondering about that. In the old days, digital imaging was something new and advanced, today, it seems like a basic introductory topic, especially if the darkroom is gone.

When would you introduce digital imaging to microscopy students and what would you include so they can be at least 'trainable' when they leave here and show up in your lab?

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Hi Leszek,

} We have to replace a HT cable in our 20 years old Philips CM-20
} SuperTwin TEM. Since a brand new cable is very expensive (what hurts
} even more because of the age of the microscope) we are looking for an
} alternative. If you know any (spare parts, microscope to be scrapped,
} etc) please let me know.

unfortunately, I only know about a CM10 and a CM12.

Unless you need that cable very urgently, you could check the web site of the German Society for Electron Microscopy (http://www.dge-homepage.de/) - there is a dedicated section for trading used equipment and a spare cable in Germany would be right around the corner.

Good luck,

Guenter

--
Dr. Guenter Resch
Nexperion e.U. - Solutions for Electron Microscopy - www.nexperion.net
Mattiellistrasse 3/17, 1040 Vienna, Austria - Phone +43 664 94 17 210
Registered at Commercial Court Vienna, FN 397677w - VAT Reg. ATU67962234

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Leszek - however counter-intuitive this may sound, but making
highly-specialized parts in single quantities for old instruments is
often more complicated (and thus expensive) then for newer models...

If you need your HV cable restored, you may try finding a local company
specializing in repairs and support of X-Ray equipment - these people
would have the right expertize, tools, and materials needed for such
job, and are quite likely to repair your cable for the lesser cost then
the new replacement.

Good luck :)

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 9/17/2013 9:00 AM, L.Kepinski-at-int.pan.wroc.pl wrote:
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} Dear Fellow Microscopists,
}
} We have to replace a HT cable in our 20 years old Philips CM-20
} SuperTwin TEM. Since a brand new cable is very expensive (what hurts
} even more because of the age of the microscope) we are looking for an
} alternative. If you know any (spare parts, microscope to be scrapped,
} etc) please let me know.
}
} With best regards,
} Leszek
}
} Leszek Kepinski
} Institute of Low Temperature and Structure Research,
} Polish Academy of Sciences,
} Okolna 2, 50-422 Wrocław, Poland
} e-mail: l.kepinski-at-int.pan.wroc.pl
}
}
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Name: Carol Heckman

Organization: Michigan Microscopy & Microanalysis

Title-Subject: [Filtered] Annual Meeting

Message: Michigan Microscopy and Microanaysis Society opens its Annual Meeting registration!

The Meeting will be hosted by Dow Chemical Corporation in Midland, MI, on Friday, October 18th. The
abstract deadline is September 27th. The day-long meeting will consist of a facility tour, a
catered lunch, vendors' displays, and technical talks. Confirmed speakers are:

Ilke Arslan
3-D and In-situ Characterization of Nanomaterials in the Scanning Transmission Electron Microscope
Dr. Arslan is Senior Scientist at the Pacific Northwest National Laboratory in Richland, Washington.
She studies the 3D nature of nanomaterials in STEM by using tomography and reconstruction techniques.

Bob Price
Some Basics of Confocal Imaging: How Deep is Deep and are They Really Colocalized?
Dr. Price is Editor-in-Chief of the MSA journal, Microscopy and Microanalysis. He will address some
common questions about confocal microscopy. How deep can I image into a tissue? Are the labeled
structures co-localized? Can I quantify the fluorescence in the images?

Stephanie Brock
Solid State and Nanomaterials
Dr. Brock has developed strategies for the synthesis of transition metal pnictides (pnicogen = Group
15 element) as nanostructures. The challenge is to assemble them into functional architectures while
retaining the unique, size-defined properties of the nanostructures.

Additional presentations will be selected from the submitted abstracts. Further details can be
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We would like to colorize �SEM images of insects to make a visual impact for an art piece for framing. We have taken both a secondary electron image and a separate back scattered image of the same field of view. We have mucked around with these images in Photoshop trying to merge them together to produce brighter highlights, changing the hue and colours but �the end result has been not very appealing.

We have also tried colorizing just using the secondary electron image but the end result has been flat looking, with less highlights.

Does anyone know how to do this successfully, any web sites, you tube videos explaining how to do this. Or are there and any software packages available to do this more easily with better results.

Thanks in advance for any suggestions, much appreciated.

Sandra Crameri
Project Leader | Diagnostic Electron Microscopy
AAHL Biosecurity Microscopy Facility
Australain Animal Health Laboratory
Animal, Food and Health Sciences
CSIRO
T +61 3 5227 5209 M +61 043 076 6643
Private Bag 24, Geelong VIC, 3220, AUSTRALIA
5 Portarlington Rd, East Geelong, Vic. 3219, AUSTRALIA

==============================Original Headers==============================
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realname - John C. Arnold
Email - John.Arnold1-at-cox.net
ORGANIZATION - Arizona State University Advanced Law Degree in Science,
Technology, Genetics and the Law
EDUCATION - Graduate College
LOCATION - Tempe, AZ 85280
SUBJECT_OF_QUESTION - Law and microscopy
QUESTION - I believe it is obvious that microscopy will become more
important in evidence law in the future. I am working on a
legal-scientific article on the foundational requirements in emerging
technologies focusing on microscopy. Is there anyone in the society who
is interested in the interrelationship of microscopy and the law?

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Hi Sandra,
David Scharf has made a business out of doing this. You could check out
http://www.scharfphoto.com/
He has used multiple detectors at different locations, in order to get the
signal differences needed to create an illumination effect with 3D
"lighting". You can see that on his website.

Good luck,
Larry Scipioni
ZS Genetics

-----Original Message-----
X-from: Sandra.Crameri-at-csiro.au [mailto:Sandra.Crameri-at-csiro.au]
Sent: Tuesday, September 17, 2013 11:41 PM
To: LES-at-ZSGENETICS.COM

We would like to colorize �SEM images of insects to make a visual impact for
an art piece for framing. We have taken both a secondary electron image and
a separate back scattered image of the same field of view. We have mucked
around with these images in Photoshop trying to merge them together to
produce brighter highlights, changing the hue and colours but �the end
result has been not very appealing.

We have also tried colorizing just using the secondary electron image but
the end result has been flat looking, with less highlights.

Does anyone know how to do this successfully, any web sites, you tube videos
explaining how to do this. Or are there and any software packages available
to do this more easily with better results.

Thanks in advance for any suggestions, much appreciated.

Sandra Crameri
Project Leader | Diagnostic Electron Microscopy AAHL Biosecurity Microscopy
Facility Australain Animal Health Laboratory Animal, Food and Health
Sciences CSIRO T +61 3 5227 5209 M +61 043 076 6643 Private Bag 24, Geelong
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Hi Sandra,

as others from the list answered, there are two principal ways to get colour in your SEM images:
- make good black and white images and add a lot of "Photoshop magic" to bring colour to the specimen
- use multiple detectors and perhaps also different kind of detectors (like SE or backscattered...) to get different signals back
from the specimen and use these signals to attribute aesthetical colours to the image (and add some Photoshop magic...).

For example: I use the secondary electrons for getting a very fast separation of parts on specimen with a small interaction volume
like hairs on insects or cells or substrate etc.
The backscattered detectors (I have three) I use to get surfaces (topography) ad hoc separated in a different colour.
You can also try to get more different images with different accelerating voltages and the best suited detectors for it...
Also distance of the detectors to the specimen can make very different iamges...

There is a PDF from a MIKROKOSMOS article which describes my work, but only in German language:
www.elektronenmikroskopie.info/pdf/Mikrokosmos06-99.pdf (1 MB)

I am using a digital image aquisition system from point electronic, www.pointelectronic.de, which can handle up to 8 detectors and
scan up to 4 detectors in a resolution of max. 16kx16k pixel. The scanning software has the ability to directly mix the signals
from the SEM into a coloured image.

For the images, have a look at:
http://www.electronmicroscopy.info/shop_biology_plants.htm for biologic specimen
and
http://www.electronmicroscopy.info/shop_materials.htm for some images from material science.
Sorry, pages are in parts still in construction.

And if you would like to see some very crazy stuff concerning colors in the SEM AND movement, see my nanoflights at
www.nanoflight.info.
For this project I got the "Technikpreis 2013" from the German Society of Electron Microscopy at the bi-annual meeting three weeks
ago. :-) This had been the film for the award: https://vimeo.com/73030307

Best wishes,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 18.09.13 05:34, schrieb Sandra.Crameri-at-csiro.au:
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} We would like to colorize �SEM images of insects to make a visual impact for an art piece for framing. We have taken both a secondary electron image and a separate back scattered image of the same field of view. We have mucked around with these images in Photoshop trying to merge them together to produce brighter highlights, changing the hue and colours but �the end result has been not very appealing.
}
} We have also tried colorizing just using the secondary electron image but the end result has been flat looking, with less highlights.
}
} Does anyone know how to do this successfully, any web sites, you tube videos explaining how to do this. Or are there and any software packages available to do this more easily with better results.
}
} Thanks in advance for any suggestions, much appreciated.
}
} Sandra Crameri
} Project Leader | Diagnostic Electron Microscopy
} AAHL Biosecurity Microscopy Facility
} Australain Animal Health Laboratory
} Animal, Food and Health Sciences
} CSIRO
} T +61 3 5227 5209 M +61 043 076 6643
} Private Bag 24, Geelong VIC, 3220, AUSTRALIA
} 5 Portarlington Rd, East Geelong, Vic. 3219, AUSTRALIA
}
}
}
}
}
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17, 23 -- From stefan.diller-at-t-online.de Thu Sep 19 07:27:42 2013
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Dear Listers
We are looking for the Jeol JSM 5600 LV column circuit diagrams. I will be
gratefull if anyone can send us these diagrams either in
electronic or in hard copy form. We are ready to cover any expences.
Thank you in advance

yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

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Dear All, may i ask if anyone is no longer using JEOL 2010F and would like to donate the airlock cover for the TEM holder/goniometer as mention in this paper

http://muller.research.engineering.cornell.edu/sites/WEELS/summer06/envirpap-print.pdf (Fig 9)

I have a JEOL 2100F of older version. Initially i wanted to purchase an airlock cover from JEOL but found out that their airlock cover is meant for the newer version of the goniometer. I also ask JEOL if i can purchase the one found on the paper above thinking that it might be compatible with our goniometer but was told JEOL no longer manufacture this old version of airlock cover

On the other hand, may i ask if anyone can point to me if there is a third party manufacturer who can customize the airlock cover for us? Would it be reliable?

Best Regards,
Yee Yan
FACTS Lab
Nanyang Technological University (Singapore)

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JEOL 100s TEM ( with biologic chamber)
JEOL isw 40 SEM
JEOL 35 SEM

all are in full working condition with manuals, extra boards...

Looking to donate to someone who can use it.

Location is near RTP, North Carolina

Thank you,
Anatoli
919.260.1911
toli-at-hillweb.com

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Hi Sandra,

You need 3 images for RGB channels or layers. It is easy if you have solid state BSE detector: you can make pictures with just one sector A, another picture with sector B, compositional picture A+B, topographical picture A-B, and, of course, SE picture. So, you will have five pictures and you need just three of them. In Photoshop try adjusting channels (layers) independently before applying operations to the whole image.

You can also use the same technique even if you have just one SE image. You can convert it in three images in various ways, for example image as is, inverted image (negative), high contrast image.

Some examples of images:
https://www.dropbox.com/sh/ddm8z2m4bhfxcqu/Ul4ZIZ4WzO?lst
Colored image of table salt was obtained from the single SE image.

Good luck,

Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: Sandra.Crameri-at-csiro.au [mailto:Sandra.Crameri-at-csiro.au]
Sent: Tuesday, September 17, 2013 10:30 PM
To: Dusevich, Vladimir

We would like to colorize �SEM images of insects to make a visual impact for an art piece for framing. We have taken both a secondary electron image and a separate back scattered image of the same field of view. We have mucked around with these images in Photoshop trying to merge them together to produce brighter highlights, changing the hue and colours but �the end result has been not very appealing.

We have also tried colorizing just using the secondary electron image but the end result has been flat looking, with less highlights.

Does anyone know how to do this successfully, any web sites, you tube videos explaining how to do this. Or are there and any software packages available to do this more easily with better results.

Thanks in advance for any suggestions, much appreciated.

Sandra Crameri
Project Leader | Diagnostic Electron Microscopy AAHL Biosecurity Microscopy Facility Australain Animal Health Laboratory Animal, Food and Health Sciences CSIRO T +61 3 5227 5209 M +61 043 076 6643 Private Bag 24, Geelong VIC, 3220, AUSTRALIA
5 Portarlington Rd, East Geelong, Vic. 3219, AUSTRALIA

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Back again, this time a question about keeping propylene oxide in the lab

We have something like 8 pints of propylene oxide in the lab, some never opened.

I think I should get rid of it, but wanted to check with the experts.

My understanding is that PO is not required as a transitional solvent, acetone works and is less toxic.

I have not used PO for a long time, but maybe there is a good reason to keep some in the lab, or should it all go?

Looking forward to your comments.

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Organization: MRL, University of Illinois

Title-Subject: [Filtered] Invitation to a Fall Conference at the University of Illinois

Message: Greetings,

We are holding our second annual Biological Conference at the Materials Laboratory on the campus of
the University of Illinois.

This is a 2 day workshop designed for students, staff, and folks crossing borders in research into
biological areas.

The conference will include a vendor fair, lectures, tours, and an open lab on the second day.
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fluorescence and more. Wolfram's Mathematica will also be presented. Tours of our Beckman, MRL
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Dear List Members,

I would like to announce the opening of two Experienced Researcher positions
(Marie Curie ERs), one for 5 months and one for 12 months, in material
characterization by TEM, at the Aristotle University of Thessaloniki,
Greece.
http://ec.europa.eu/euraxess/index.cfm/jobs/jobDetails/33880551

Best regards,
E.K. Polychroniadis

Prof. E.K. Polychroniadis
Department of Physics
Aristotle University of Thessaloniki
Thessaloniki 54124, Greece
Tel.: +30.2310.998163
Fax: +30.2310.998241
e-mail: polychr-at-auth.gr

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1. Is there any available info on calibrating the column? I've noticed that when trying to beam shift to a new center point, the resulting image is substantially offset from what should have been the center. I'd also tried to image something that I thought I knew the dimensions of, and saw a significant error when I used the software to measure it. I'd thought I made a mistake, but in light of the beam shift issue, I'm not so sure. I'm considering getting a real calibration standard, but I don't have any service docs for calibrating the column.

2. I had thought filament emission was supposed to have a first peak and then a second plateau. I've seen that before, but lately now all I can find is the first peak. Raising the filament current, I see a first peak, at a little over 2.4A, then a fall off, and no further increase in brightness even though I increase the current by up to another amp. That doesn't seem right. When I've seen a second peak/plateau in the past, I would always see an increase begin fairly soon after the first peak as I raised filament current. Any thoughts?

3. I'm not sure OptiBeam is recommending apertures like it should. Usually I see recommendations to change fairly infrequently, and often during intermittent conditions like after startup. But when I go to the Apertures window, I see the current aperture recommended as best. Whenever I change apertures, invariably the suggested aperture changes to match the new aperture.

Now the Apertures dialog has three numeric windows below the aperture selection: Defining Aperture, Left, Right, and a checkbox for Film Aperture. I have no idea what these are as I don't see them mentioned in the documentation, and I wonder if an error here might be the problem.

4. The SEM has an HRRU. What exactly makes it "high resolution"? I see it is connected to the SEM by fiber optic cables, so I'm assuming it uses a digital signal. Does it just get the 1024x768x8 bit signal that the display gets? Is the resulting Polaroid image any better than what you would see in the screen or could print from a saved tif file?

Thanks in advance, and thanks to those who helped with my previous questions.

Ryan Pierce
1. Is there any available info on calibrating the column? I've noticed that when trying to beam shift to a new center point, the resulting image is substantially offset from what should have been the center. I'd also tried to image something that I thought I knew the dimensions of, and saw a significant error when I used the software to measure it. I'd thought I made a mistake, but in light of the beam shift issue, I'm not so sure. I'm considering getting a real calibration standard, but I don't have any service docs for calibrating the column.

2. I had thought filament emission was supposed to have a first peak and then a second plateau. I've seen that before, but lately now all I can find is the first peak. Raising the filament current, I see a first peak, at a little over 2.4A, then a fall off, and no further increase in brightness even though I increase the current by up to another amp. That doesn't seem right. When I've seen a second peak/plateau in the past, I would always see an increase begin fairly soon after the first peak as I raised filament current. Any thoughts?

3. I'm not sure OptiBeam is recommending apertures like it should. Usually I see recommendations to change fairly infrequently, and often during intermittent conditions like after startup. But when I go to the Apertures window, I see the current aperture recommended as best. Whenever I change apertures, invariably the suggested aperture changes to match the new aperture.

Now the Apertures dialog has three numeric windows below the aperture selection: Defining Aperture, Left, Right, and a checkbox for Film Aperture. I have no idea what these are as I don't see them mentioned in the documentation, and I wonder if an error here might be the problem.

4. The SEM has an HRRU. What exactly makes it "high resolution"? I see it is connected to the SEM by fiber optic cables, so I'm assuming it uses a digital signal. Does it just get the 1024x768x8 bit signal that the display gets? Is the resulting Polaroid image any better than what you would see in the screen or could print from a saved tif file?

Thanks in advance, and thanks to those who helped with my previous questions.

Ryan Pierce

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Hi Ryan

I am a little rusty on the 430 now but Let us look at the problems that you
have and try to find an explanation.

1. Calibrating the column is a difficult title, calibrating the
magnification is a sensible act to follow. Instruments are usually within 5
to 10% of the readout but there are a number of areas that may introduce
errors that are capable of increasing these errors. Buy yourself a
calibration grating either for a TEM (yes a transmission electron
microscope) which will be 10s of $ or if you are rich you may buy a SEM
grating calibration standard. The best way to learn is to check things out
for yourself!
2. This area worries me because what you explain suggests that the
electron gun is not correctly aligned. Let's look at practical experience.
As you run up the filament heating the source starts at a pretty large size,
but it reduces in size through the action of the heat and the bias circuit.
So when you start heating the filament the source is so large that you have
electrons being passed down the column. When you apply more heat the source
becomes smaller and may no longer be centred on the column and the signal
drops. Take a 25 cent coin and place a 1 cent coin onto it, but touching
one edge. Your low heat beam is the 25 cent coin but the 1 cent coin
represents your high heat situation. The beam is off centre and the more
you heat up the filament the smaller the source and the more the
misalignment. To correct the problem heat up your filament until you have no
change or a drop in signal level and then gently adjust the gun tilt
controls to increase the signal level. You could try the gun shift as well
but this should have minimal effect if reasonably well aligned. Heat the
filament still further and repeat the tilt alignment. The gun is fully
aligned when the signal intensity does not change with an increase in
filament current but do not go too near to a 2.8amp maximum. Checking the
procedure you should now find that you do have a false peak before the main
peak?
3. Don't tie yourself in knots trying to understand OptiBeam. These are
gross settings for particular applications but operating the instrument in
almost any mode should give you reasonable results. What do most operators
require? (a) A signal good enough to operate with ease (b) with a spot size
(probe current or I probe) that enables a good quality image to be produced.
There are manipulations with aperture sizes, working distance, kV etc etc
but why bother if you have answered "a" or "b" above? It is normal to
change an aperture for a gross change in probe current, up or down, other
more subtle probe current adjustments are accomplished through the spot size
or probe current controls. However changing the physical aperture on many
instruments re calibrates the probe current values so staying with one
aperture it is easier to understand what you are achieving with the spot
size (probe current). At this stage may I suggest that you always work with
regard to probe current as this terminology is used by other manufacturers
and this makes it easier to move from instrument to instrument.
4. The HRRU is what is says it is a High Resolution Record Unit, in
other words a high resolution CRT that is being used specifically for old
fashioned photography. This is a hard wired system using a smaller higher
quality display than that used by the operator. The world has argued about
photography and digital images and I think we are all now very happy with
our digital images but of course you must compare like with like, the number
of lines on your photograph and the number of lines depicted on your digital
image.

I hope that this helps but for further information take a look at "Hints and
Tips" on my web site.

Regards

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

----------------------------------------------------------------------------
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1. Is there any available info on calibrating the column? I've noticed that
when trying to beam shift to a new center point, the resulting image is
substantially offset from what should have been the center. I'd also tried
to image something that I thought I knew the dimensions of, and saw a
significant error when I used the software to measure it. I'd thought I made
a mistake, but in light of the beam shift issue, I'm not so sure. I'm
considering getting a real calibration standard, but I don't have any
service docs for calibrating the column.

2. I had thought filament emission was supposed to have a first peak and
then a second plateau. I've seen that before, but lately now all I can find
is the first peak. Raising the filament current, I see a first peak, at a
little over 2.4A, then a fall off, and no further increase in brightness
even though I increase the current by up to another amp. That doesn't seem
right. When I've seen a second peak/plateau in the past, I would always see
an increase begin fairly soon after the first peak as I raised filament
current. Any thoughts?

3. I'm not sure OptiBeam is recommending apertures like it should. Usually I
see recommendations to change fairly infrequently, and often during
intermittent conditions like after startup. But when I go to the Apertures
window, I see the current aperture recommended as best. Whenever I change
apertures, invariably the suggested aperture changes to match the new
aperture.

Now the Apertures dialog has three numeric windows below the aperture
selection: Defining Aperture, Left, Right, and a checkbox for Film Aperture.
I have no idea what these are as I don't see them mentioned in the
documentation, and I wonder if an error here might be the problem.

4. The SEM has an HRRU. What exactly makes it "high resolution"? I see it is
connected to the SEM by fiber optic cables, so I'm assuming it uses a
digital signal. Does it just get the 1024x768x8 bit signal that the display
gets? Is the resulting Polaroid image any better than what you would see in
the screen or could print from a saved tif file?

Thanks in advance, and thanks to those who helped with my previous
questions.

Ryan Pierce

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Thanks for the info.

1. Regarding calibration, I know I can determine the percent error for myself between the grid size and on screen measurement, although I am not sure this would stay the same at different magnifications, kV settings, or working distances. I was wondering if there were a way to input this into the scope so it could display the correct info. It has been a while since I imaged something of known dimensions but I seem to recall getting a number substantially more than 10% error.

This is a hackerspace with some seriously skilled people in electronics and instrument calibration. If we had the procedures, and assuming we could save the old calibration data to recover from in case we made it worse, it would be something worth trying. The question is whether service guides for the scope that document the procedure could be obtained, and whether specialized tools and test equipment is needed.

2. This is really interesting. I didn't realize the filament changes size like that, and what you say about it shrinking and moving off center makes sense. But we don't have gun tilt. The only things we can change on an S430 are: The four screws centering the filament inside the Wehnelt. The nut adjusting the height of the filament relative to the Wehnelt. The four knobs on the outside of the column that adjust the gun position relative to the column. I can try adjusting those tonight to see if I can get more intensity and a second peak. Otherwise I guess I'll need to take the gun apart and check it.

4. I know Polaroid is outdated and that specific type of film might be unavailable. I also know the image processor board had a 1024x768x8 bit resolution, and that is what I see when I save a TIF file, or what I see on the screen. But the connection to the HRRU is digital. If it is just the same 1024x768 image that I'm seeing now, it is not of interest to me. But if the image processor board is sending data at a higher resolution to the HRRU, then it may be possible to build my own data capture system that pretends to be the HRRU, listens to the digital data, and stores it as higher resolution files than the scope produces. Hence my interest in just how high the resolution of the HRRU is.

One other practical question:

I'd heard that a SEM column should always stay under vacuum except when changing samples. So I've been keeping the pumps running 24x7. Is that the best thing to do? Our vacuum seems to stay between 6E-5 and 9E-5 torr which doesn't seem as good as I'd like, but I don't know where to start should I try replacing gaskets, etc. or if it could be a pump problem.

Thanks,
Ryan

} On Sep 25, 2013, at 4:51 AM, protrain-at-emcourses.com wrote:
}
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
} To Subscribe/Unsubscribe -- http://www.microscopy.com/MicroscopyListserver
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} ----------------------------------------------------------------------------
}
} Hi Ryan
}
} I am a little rusty on the 430 now but Let us look at the problems that you
} have and try to find an explanation.
}
} 1. Calibrating the column is a difficult title, calibrating the
} magnification is a sensible act to follow. Instruments are usually within 5
} to 10% of the readout but there are a number of areas that may introduce
} errors that are capable of increasing these errors. Buy yourself a
} calibration grating either for a TEM (yes a transmission electron
} microscope) which will be 10s of $ or if you are rich you may buy a SEM
} grating calibration standard. The best way to learn is to check things out
} for yourself!
} 2. This area worries me because what you explain suggests that the
} electron gun is not correctly aligned. Let's look at practical experience.
} As you run up the filament heating the source starts at a pretty large size,
} but it reduces in size through the action of the heat and the bias circuit.
} So when you start heating the filament the source is so large that you have
} electrons being passed down the column. When you apply more heat the source
} becomes smaller and may no longer be centred on the column and the signal
} drops. Take a 25 cent coin and place a 1 cent coin onto it, but touching
} one edge. Your low heat beam is the 25 cent coin but the 1 cent coin
} represents your high heat situation. The beam is off centre and the more
} you heat up the filament the smaller the source and the more the
} misalignment. To correct the problem heat up your filament until you have no
} change or a drop in signal level and then gently adjust the gun tilt
} controls to increase the signal level. You could try the gun shift as well
} but this should have minimal effect if reasonably well aligned. Heat the
} filament still further and repeat the tilt alignment. The gun is fully
} aligned when the signal intensity does not change with an increase in
} filament current but do not go too near to a 2.8amp maximum. Checking the
} procedure you should now find that you do have a false peak before the main
} peak?
} 3. Don't tie yourself in knots trying to understand OptiBeam. These are
} gross settings for particular applications but operating the instrument in
} almost any mode should give you reasonable results. What do most operators
} require? (a) A signal good enough to operate with ease (b) with a spot size
} (probe current or I probe) that enables a good quality image to be produced.
} There are manipulations with aperture sizes, working distance, kV etc etc
} but why bother if you have answered "a" or "b" above? It is normal to
} change an aperture for a gross change in probe current, up or down, other
} more subtle probe current adjustments are accomplished through the spot size
} or probe current controls. However changing the physical aperture on many
} instruments re calibrates the probe current values so staying with one
} aperture it is easier to understand what you are achieving with the spot
} size (probe current). At this stage may I suggest that you always work with
} regard to probe current as this terminology is used by other manufacturers
} and this makes it easier to move from instrument to instrument.
} 4. The HRRU is what is says it is a High Resolution Record Unit, in
} other words a high resolution CRT that is being used specifically for old
} fashioned photography. This is a hard wired system using a smaller higher
} quality display than that used by the operator. The world has argued about
} photography and digital images and I think we are all now very happy with
} our digital images but of course you must compare like with like, the number
} of lines on your photograph and the number of lines depicted on your digital
} image.
}
} I hope that this helps but for further information take a look at "Hints and
} Tips" on my web site.
}
} Regards
}
} Steve
}
} Steve Chapman FRMS
} Protrain for Consultancy and Courses in Electron Microscopy
} Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
} www.emcourses.com
}

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OK, call me naive, but I have a question about getting formvar films to release.

Students here are going through the initiation process of making coated grids. Sometimes it works, sometimes it doesn't, even for me.

I was looking over a Dear Abbe column in a recent Microscopy Today that addressed this same question. It suggested a few tricks I had never heard of.

One that was intriguing was to lick the slide first. Now I had never heard of that, but am getting desperate enough to try it.

I know some of our students might read this advice from Abbe and try it without clearing it, is it for real?

Seriously, how about an update on any tips or tricks on getting that formvar off the slide every time.

Thanks

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Jon,
You have to sacrifice an ingot of osmium to the goddess of adhesion...

Don't laugh but here is my, highly reliable if not completely
perfect, superstitious ritual.

Obtain Ross Optial Lens Tissue. This is important, accept no substitutes.

Take a piece of lens tissue and vigorously rub the slide surface to
be coated. To make it less stressful, I rub both sides of the slide
so I don't have to keep track. I typically count ten back and forths
(20 each way). Then flip the slide and do another 10.

Then dip in Formvar, let dry, score, and float.

Having written this down, no doubt the next time I try, it won't
work. But up until then, this method has absolutely lowered my stress
level for this task.

As ever
Tobias
}
} OK, call me naive, but I have a question about getting formvar films
} to release.
}
} Students here are going through the initiation process of making
} coated grids. Sometimes it works, sometimes it doesn't, even for me.
}
} I was looking over a Dear Abbe column in a recent Microscopy Today
} that addressed this same question. It suggested a few tricks I had
} never heard of.
}
} One that was intriguing was to lick the slide first. Now I had never
} heard of that, but am getting desperate enough to try it.
}
} I know some of our students might read this advice from Abbe and try
} it without clearing it, is it for real?
}
} Seriously, how about an update on any tips or tricks on getting that
} formvar off the slide every time.
}
} Thanks
}
} Jon
}
} Jonathan Krupp
} Applied Science, Business & Technology
} San Joaquin Delta College
} 5151 Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu
}
} Find us on Facebook -at-
} Electron Microscopy at SJ Delta College
}
}

--
_ ____ __ ____
/ \ / / \ / \ \ Tobias I. Baskin
/ / / / \ \ \ Professor
/_ / __ /__ \ \ \__ Biology Department
/ / / \ \ \ 611 N. Pleasant St.
/ / / \ \ \ University of
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/ / ___ / \ \__/ \ ____ Amherst, MA, 01003
www.bio.umass.edu/biology/baskin
Voice: 413 - 545 - 1533 Fax: 413 - 545 - 3243

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Jon,

We went through this same issue a few years ago with our TEM class:
cleaning slides, not cleaning slides, clean but leave a bit of
"soapiness", breathing on the coated slides, nose grease, squashed
cochroach goo (hey, it's cheap), etc. etc., and eventually settled on
one thing:
Grafco Cat # 3703-2P slides with clear/ground edges, Graham-Field Health
Products www.grahamfield.com .
They're cheap and reliably release the formvar film. Found them in the
Intro Biol lab supplies, because they're cheap. The other brands &
catalog number slides we tried had the same issues you're having.

Phil
P.S. "You're naive."

} OK, call me naive, but I have a question about getting formvar films to release.
}
} Students here are going through the initiation process of making coated grids. Sometimes it works, sometimes it doesn't, even for me.
}
} I was looking over a Dear Abbe column in a recent Microscopy Today that addressed this same question. It suggested a few tricks I had never heard of.
}
} One that was intriguing was to lick the slide first. Now I had never heard of that, but am getting desperate enough to try it.
}
} I know some of our students might read this advice from Abbe and try it without clearing it, is it for real?
}
} Seriously, how about an update on any tips or tricks on getting that formvar off the slide every time.
}
} Thanks
}
} Jon
}
} Jonathan Krupp
} Applied Science, Business& Technology
} San Joaquin Delta College
} 5151 Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Hi Jon,

Because casting films onto microscope slides is a"black art", I use
freshly leaved mica sheets for both carbon and formvar films. I just
dip the mica into the solution and let it dry. It is necessary to take
a razor to score the edges so that the water can penetrate under the
film but once it does, the films lift right off... no black magic or
voodoo chants required!

Good luck,
Henk

At 9/25/2013 1:23 PM, jkrupp-at-deltacollege.edu wrote:
}
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} OK, call me naive, but I have a question about getting formvar films to release.
}
} Students here are going through the initiation process of making coated grids. Sometimes it works, sometimes it doesn't, even for me.
}
} I was looking over a Dear Abbe column in a recent Microscopy Today that addressed this same question. It suggested a few tricks I had never heard of.
}
} One that was intriguing was to lick the slide first. Now I had never heard of that, but am getting desperate enough to try it.
}
} I know some of our students might read this advice from Abbe and try it without clearing it, is it for real?
}
} Seriously, how about an update on any tips or tricks on getting that formvar off the slide every time.
}
} Thanks
}
} Jon
}
} Jonathan Krupp
} Applied Science, Business & Technology
} San Joaquin Delta College
} 5151 Pacific Ave.
} Stockton, CA 95207
} 209-954-5284
} jkrupp-at-deltacollege.edu
}
} Find us on Facebook -at-
} Electron Microscopy at SJ Delta College
}
}
}
}
}
}
}
}
}
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--
Untitled Document

Hendrik O. Colijn
*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*
1305 Kinnear Rd, Suite 100, Columbus, OH 43212

614.643.3458 Office colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu}
www.cemas.osu.edu {http://www.cemas.osu.edu}

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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Listers,

Not my reply, but forwarded for an industrial microscopist who must
remain anonymous:
"When I made my own films many years ago, I used the bevel-edge slides
you described. An important step is to etch a rectangle in the Formvar
film on the slide prior to releasing it onto the surface of the water.
I did this by using a sharp razor blade and cutting several millimeters
inside the edges of the slide. Others in my lab scraped the edge of the
slide with a razor blade to promote release."

Aren't lawyers fun? A person can't reveal their name for a post like
this ...

Phil
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Hey Jon,

In a previous life we would meticulously clean our slides (which "had" to be a certain brand, but I don't remember which one), then place them in racks in a vacuum evaporator with a big bell jar. We would use a tungsten coil and place a chunk of a soapy substance called Victawet in the basket, then pump down and heat up the coil until all the Victawet evaporated. Then we took out the coated slides, which looked like they had a hazy film on them, and box them up until needed. Before dipping the slides in formvar solution we would polish them vigorously again. The Victawet seemed to act like a release agent, but sometimes even that didn't work.

We had this long elaborate ritual with uneven results. Years later I mentioned this to Kent MacDonald and he proceeded to demonstrate his method, which was a no-frills, straightforward "pick a slide out of the box, wipe it off, dip it in formvar and dip it in water" sequence. Beautiful films which released the first time. I seem to recall that he polished the slide with his shirt, but I'm not sure anymore, and I beg his forgiveness if I have misremembered that part!

Motto: formvar film release appears to be in the hands of the gods (and EM legends like Kent).

Respectfully,

Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211

Phone: 573-884-5383
Email: tindallr-at-missouri.edu
Day job: www.emc.missouri.edu
Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
Blog: http://nadiasyard.com

-----Original Message-----
X-from: jkrupp-at-deltacollege.edu [mailto:jkrupp-at-deltacollege.edu]
Sent: Wednesday, September 25, 2013 12:34 PM
To: Tindall, Randall D.

OK, call me naive, but I have a question about getting formvar films to release.

Students here are going through the initiation process of making coated grids. Sometimes it works, sometimes it doesn't, even for me.

I was looking over a Dear Abbe column in a recent Microscopy Today that addressed this same question. It suggested a few tricks I had never heard of.

One that was intriguing was to lick the slide first. Now I had never heard of that, but am getting desperate enough to try it.

I know some of our students might read this advice from Abbe and try it without clearing it, is it for real?

Seriously, how about an update on any tips or tricks on getting that formvar off the slide every time.

Thanks

Jon

Jonathan Krupp
Applied Science, Business & Technology
San Joaquin Delta College
5151 Pacific Ave.
Stockton, CA 95207
209-954-5284
jkrupp-at-deltacollege.edu

Find us on Facebook -at-
Electron Microscopy at SJ Delta College

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Listers,
Well, the fates clearly have it in Formvar. Moments after
posting about the miraculous Ross Optical Lens Tissue (and I use the
word miraculous in all seriousness), I was informed by Ted Pella (who
sold me my treasured box of Ross) that these tissues are no longer
made. He tells me that the replacement product their company found
has been field tested in EM labs and found satisfactory. But given
the notorious quirkiness of Formvar, one has to wonder...

One good thing from Ted Pella about the replacement--the new
box really does have 1000 tissues--apparently the Ross box claimed
1000 but actually had only 800 (they counted!). And despite this
dodge Ross went out of business. Perhaps a warning?

Formvar and prosper,
Tobias

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Email: andrei.kolmakov-at-nist.gov
Name: Andrei Kolmakov

Organization: NIST CNST

Title-Subject: [Filtered] postdoctoral position available

Message: Dear Colleagues,
Could you please pass the following information to the attention of
appropriate candidates in your group and/or beyond?

NIST Center for Nanoscale Science and Technology is expanding its
research program toward development of in situ electron microscopy and
spectroscopy of objects and devices in liquids and dense gaseous
environments using environmental cells equipped with graphene based
electron transparent windows. Currently, a postdoctoral position for
highly motivated and experienced experimentalist is available. The
preferable set of skills includes but not limited: SEM, XPS, AES, UHV
surface science protocols, clean room experience, graphene transfer
protocols, graphene functionalization , electrochemistry, excellent
writing and teamwork skills and etc. The application letter, CV with
publication record and contact names of 3-4 references should be sent to
Dr. Andrei Kolmakov, andrei.kolmakov-at-nist.gov

Thank you in advance

Andrei Kolmakov

Project Leader CNST NIST Energy Group
NIST 100 Bureau Drive
Bldg. 216/Rm.B117
Gaithersburg, MD 20899-6204
Phone: (301) 975-4724
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Email: smythen-at-musc.edu
Name: Nancy Smythe

Organization: Medical University of South Carolina

Title-Subject: [Filtered] diamond knives

Message: We have been using the same brand of diamond knife for 30 years
but in the last few years the quality seems to have declined. When we
get one that is scratch free it doesn't last more than a few months. Is
anyone happy with their knife company? Please drop me a line and tell me
the company.

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Nancy,

A very wise lady (and owner of a large EM supply company) once told me to
check my microtomes when I had a similar problem. I had not had them
serviced for quite a while. Once they were serviced the problem
disappeared. Based on that experience I would recommend you having them
serviced every 1-2years based on usage. It does cost to do so but it is
much less than resharpening knives.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Email: johns248-at-miamioh.edu
Name: Christina Johnson

Organization: Miami University

Title-Subject: [Filtered] Slide Storage

Message: Hello List Members,

What is the best way to store samples that have been stained with
Toluidine blue?

I am working with semi-thin sections of LR white-embedded plant
(*Arabidopsis*) tissue. They are mounted on glass slides with permount,
stained in Tol Blue. I currently have them organized in slide boxes on
a shelf in my office, which is consistently at or below 70 degrees
Fahrenheit. What more can I do to ensure their longevity? I would like
to make sure that the staining will last years of storage.

Thank you for your time!
Christina Johnson

PhD Candidate
Center for Advanced Microscopy & Imaging
Miami University

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I know there is not many of these instruments around, but wonder if somebody has experience with sample preparation for these. Zeiss recommended procedure calls for aldehyde fixation, post-fixation with reduced osmium tetroxide, liganding with thiocarbazide, uranyl acetate and lead aspartate staining. The sample is mounted using conductive cement and sputter coated with gold from the sides.

We got some reasonable results with this protocol, but charging is still an issue. Any advice how to make the biological (tissue or cell pellet) sample more conductive?

Thanks,

Michal Jarnik, Ph.D.
Electron Microscope Facility
Cell Biology and Metabolism Program
National Institute of Child Health and Human Development
National Institutes of Health
9000 Rockville Pike, Bldg. 18T, Room 101
Bethesda, MD 20892

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realname - Allison Boley
Email - Allison.Boley-at-asu.edu
ORGANIZATION - Arizona State University
EDUCATION - Graduate College
LOCATION - Tempe, AZ, United States
SUBJECT_OF_QUESTION - Wanted: Used TEM Sample Prep Equipment
QUESTION - I'm having technical problems submitting this to the
listserve directly, so perhaps this avenue will prove more successful.

Our group is looking to acquire two pieces of TEM sample prep equipment:

1. Gatan stereo microscope part #656.022 (used for the alignment of
Model 656 Dimple Grinder)

2. Buehler Isomet Low Speed saw or equivalent precision diamond wafering saw

If you have such new or gently used equipment that you are willing to
sell/ donate, please contact me at Allison.Boley-at-asu.edu

Thanks,
Allison Boley

Ph.D. Candidate
Department of Physics
Arizona State University
Allison.Boley-at-asu.edu

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What would be the best microscope for malaria screening in rural African
communities?
They likely have the funding sources, so that shouldn't be a concern.

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X-from: Sharon DeMoss [mailto:sharondemoss53-at-gmail.com]
Sent: Thursday, September 26, 2013 2:34 PM

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ealname - Victor Ivan Hernandez
Email - victorivanhernandez-at-kemet.com
ORGANIZATION - Kemet Electronics inc.
EDUCATION - Graduate College
LOCATION - Monterrey
SUBJECT_OF_QUESTION - Gauge R&R
QUESTION - Hello, My name is Victor Hernandez. I am a fellow
Microscopist soon to enter a masters degree of materials science.
recently i participated in a meeting where i was asked about performing
a Gauge R&R or MSA on the SEM regarding chemical analysis (EDS) i was
hoping if you might be able to help with how i may go about doing this
study and what would i need?

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Sharon, Fraen in Italy makes a nice LED microscope that can be used for fluorescence work. It can be battery powered if necessary. The lamps are LED's so do not require the maintenance that an arc lamp would need, and are much less expensive.

I used to work for a company that sold these and they were a pretty nice microscope. Not research grade but perfect for your application.

Website:
http://www.fraensrl.com/flmicro.html

Tim Morken
Supervisor, Electron Microscopy/Neuromuscular Special Studies Department of Pathology UC San Francisco Medical Center
505 Parnassus Ave, Box 1656
Room S570
San Francisco, CA 94143
�
(415) 353-1266 (ph)
(415) 514-3403 (fax)
tim.morken-at-ucsfmedctr.org

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Friday, September 27, 2013 10:07 AM
To: Morken, Timothy

What would be the best microscope for malaria screening in rural African communities?
They likely have the funding sources, so that shouldn't be a concern.

***************************************************************************************
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X-from: Sharon DeMoss [mailto:sharondemoss53-at-gmail.com]
Sent: Thursday, September 26, 2013 2:34 PM

Hi all,

We have a Jeol JEM-1200 EX Transmission electron microscope with all
the parts in full working condition with manual, amt v 542 digital
camera�.
Looking to donate to someone who can use it.
We are located in Connecticut.

Thanks,

Hilal

**********************************
Hilal Ilarslan, Ph.D

************************************

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Email: longf-at-me.queensu.ca
Name: Fei

Organization: Queen's University

Title-Subject: [Filtered] CM20 front window glass

Message: We are looking for a used front large window glass to replace the damaged one in our TEM.
The model is: NORAN
7720B-4NES-SN NANOTRACE PHILIPS CM 20T.
If anybody is deposing of such one, please let me know the price of purchasing.
Thanks a lot!

Fei

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} We have been doing TEM for years on cells growing on cell culture plates
} by doing all of the processing & embedding directly in the dish. We
} leave a little film of resin behind from the final straight resin change
} and then invert Beem capsules filled
} with resin onto the dish. After polymerization of 24 hours we then let
} them cool and "pop off" the Beem capsules bringing the cell layer with
} it. We have found through the years that certain cell culture plates
} will not stand up to the chemicals in the resin
} and you will get kind of a cloudy mess. We've also learned that any
} alcohol must be thoroughly removed before adding any resin as the
} combination seems to really eat away at the plastic in the culture
} dishes. For the same reason we don't do a propylene oxide
} wash for this procedure. However, through trial and error we have been
} able to keep a list compiled of which plates work with which resins and
} have had success using this method. We still routinely check the plates
} with the resin we intend to use before we
} do a real processing to make sure that nothing has changed since the
} last time we did it. Using this method we have always been able to get a
} resin surface where 50-75 % is usable for sectioning. There is almost
} always a small area where you might get some
} plastic that comes off with the resin.
}
}
} We have always been able to tell if the embedding worked before even
} popping the Beem capsules off by just looking at the polymerized resin.
} If it is clear the capsules will pop off mostly clean, if the resin is
} cloudy the procedure is a bust. However,
} recently we have been getting way too many situations where the resin is
} perfectly clear yet the Beem capsules bring up a fine layer of plastic
} with them that makes sectioning impossible. This seems to be happening
} with different resins and plates from different
} manufacturers. Does anyone else use this procedure for TEM of cultured
} cells and if so are there tips or tricks that would help us?
}
}
} I know you can scrape and embed in agar, etc. but I would rather continue
} doing things the way we have in the past if possible.
}
}
} Thanks for all of the replies in advance,
}
}
} Jim
}
} Jim Begley
} Research Facility Manager
} University of Kentucky Imaging Facility
} 001 HSRB
} 1095 VA Drive
} Lexington, KY 40536-0305
} Phone: 859-323-6108
} Fax: 859-323-8089
} E-mail: jgbegl2-at-uky.edu
}
}
}
}
}

==============================Original Headers==============================
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5, 28 -- Subject: FW: TEM of cells growing on cell culture plates
5, 28 -- Thread-Topic: TEM of cells growing on cell culture plates
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http://www.microscopy-uk.org.uk/mag/imgjan09/The-Model-H-Revisited.pdf� d[the old standard - gone!!!]

http://www.med-chem.com/pages/lab.../pdf/determination_of_parasitemia.pdf�

https://www.talcuk.org/accessories/microscopy-of-tropical-diseases---learning-bench-aid-series-4th-edition.htm

http://www.millennium-microscope.org/ [The new standard - beginning!]

Cheers,

Fred Monson

Frederick C Monson, PhD
Center for Microanalysis and Imaging, Research and Training (CMIRT) West Chester University of Pannsylvania West Chester, PA, 1938
610-738-0437
fmonson-at-wcupa.edu
________________________________________
X-from: oshel1pe-at-cmich.edu [oshel1pe-at-cmich.edu]
Sent: Friday, September 27, 2013 1:03 PM
To: Monson, Frederick

What would be the best microscope for malaria screening in rural African communities?
They likely have the funding sources, so that shouldn't be a concern.

***************************************************************************************
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X-from: Sharon DeMoss [mailto:sharondemoss53-at-gmail.com]
Sent: Thursday, September 26, 2013 2:34 PM

Jim,

Nearly all tissue culture dishes and flasks are made of polystyrene and I
have had good luck using my main Epon component from Ladd. It is called
LX-112. It works as well as original Epon812 in the dishes. The NMA,
DDSA, and DMP-30 I have not had a problem in using from different
suppliers. I believe that the Epon from SPI (and maybe others) will also
work but I personally have not used it.

Electron Microscopy Sciences states in their catalogue that "Embed 812 may
cause etching on selected plastics". It melts the surface of TC dishes
from at least 4 suppliers that I had checked out many years back when I
ran out of Shell's Epon812. I had informed EMS of the problem and hence
the warning. I do use their other components.

Most cells are happy growing on Permanox dishes. They are larger than I'd
wish being 60X15mm and are expensive as dishes go but one can use any
embedding chemicals as well as propylene oxide and they stay very clear
and smooth. The Beem capsules come off very easily. I requested a few
years ago for the company to make small dishes out of Permanox but I'd
need to order a run of many thousands of cases!

When it is necessary to limit the growing area as in pre-embedding immune
work we use the 2 well Permanox slides. I can fit either 2 normal sized
capsules in each well or 3 small ones. I have sliced off the end of the
capsules and filled them from the top instead of your method of filling
the capsule first. I usually put the capsules into the chamber then cure
overnight before the fill and label steps so that the Epon does not leak
out and fill the whole chamber. Within the small space of the chamber your
way may not work too well.
Note: the sides of the wells are polystyrene so do not use Propylene oxide
and they will melt if not using the LX-112 mentioned above.

Opinions and experiences related are those of Pat Connelly and do not
represent the NIH. I have no financial interest in any company mentioned I
am just a user.

Patricia Connelly
Research Assistant
NHLBI Electron Microscopy Core
National Institutes of Health
14 Service Road West
Bldg. 14E � Rm. 111B MSC 5570
Bethesda, MD 20892-5570
Phone 301-496-3491
FAX 301-402-0170
connellyps-at-nhlbi.nih.gov

This message is not confidential and can be freely shared and reproduced.
=============

{jgbegl2-at-email.uky.edu} wrote:
}
} } We have been doing TEM for years on cells growing on cell culture plates
} } by doing all of the processing & embedding directly in the dish. We
} } leave a little film of resin behind from the final straight resin change
} } and then invert Beem capsules filled
} } with resin onto the dish. After polymerization of 24 hours we then let
} } them cool and "pop off" the Beem capsules bringing the cell layer with
} } it. We have found through the years that certain cell culture plates
} } will not stand up to the chemicals in the resin
} } and you will get kind of a cloudy mess. We've also learned that any
} } alcohol must be thoroughly removed before adding any resin as the
} } combination seems to really eat away at the plastic in the culture
} } dishes. For the same reason we don't do a propylene oxide
} } wash for this procedure. However, through trial and error we have been
} } able to keep a list compiled of which plates work with which resins and
} } have had success using this method. We still routinely check the plates
} } with the resin we intend to use before we
} } do a real processing to make sure that nothing has changed since the
} } last time we did it. Using this method we have always been able to get a
} } resin surface where 50-75 % is usable for sectioning. There is almost
} } always a small area where you might get some
} } plastic that comes off with the resin.
} }
} }
} } We have always been able to tell if the embedding worked before even
} } popping the Beem capsules off by just looking at the polymerized resin.
} } If it is clear the capsules will pop off mostly clean, if the resin is
} } cloudy the procedure is a bust. However,
} } recently we have been getting way too many situations where the resin is
} } perfectly clear yet the Beem capsules bring up a fine layer of plastic
} } with them that makes sectioning impossible. This seems to be happening
} } with different resins and plates from different
} } manufacturers. Does anyone else use this procedure for TEM of cultured
} } cells and if so are there tips or tricks that would help us?
} }
} }
} } I know you can scrape and embed in agar, etc. but I would rather continue
} } doing things the way we have in the past if possible.
} }
} }
} } Thanks for all of the replies in advance,
} }
} } Jim
} }
} } Jim Begley
} } Research Facility Manager
} } University of Kentucky Imaging Facility
} } 001 HSRB
} } 1095 VA Drive
} } Lexington, KY 40536-0305
} } Phone: 859-323-6108
} } Fax: 859-323-8089
} } E-mail: jgbegl2-at-uky.edu

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Hi, Jim-

Like Pat Connelly, I use LX112 from Ladd as well. I embed in culture
dishes, without the BEEM capsules. After polymerization I just cut out the
cells of interest in a shape that fits in the ultramicrotome, or I Krazy
Glue them to blocks.

To separate the polymerized LX112 from the dish I either try to peel it
off while the dish is still hot from the embedding oven, or I go the other
route and try to separate them after dipping in liquid nitrogen (sometimes
by throwing them against the wall). It's not always pretty, but I've never
failed to get the plastic off.

Do not use propylene oxide! But LX112 in ethanol does not melt the dish or
make for foggy resin.

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*
****************************************************************************

==============================Original Headers==============================
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Dear Microscopy Community,

Recently we have purchased an EBSD detector for our Philips/FEI XL-30 SFEG
SEM in order to map nanocrystalline/ultra-fine grained materials. A problem
we are consistently running into is a shift in the map occurring
approximately every 40 minutes. This is most noticeable when attempting to
map the nanocrystalline grains with step sizes on the order of 10-50 nm.
Attached you will find a PDF documenting this issue and some of the steps we
have taken to resolve this issue, to no avail. We gratefully ask for your
help in diagnosing this issue, especially if you have run into a similar
issue before.�

Thank you very much and best regards.

Brandon Saller
3rd year Materials Science Ph.D. student
University of California, Davis

==============================Original Headers==============================
8, 25 -- From fahayes-at-ucdavis.edu Mon Sep 30 21:26:22 2013
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8, 25 -- Subject: FW: EBSD artifact
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Thanks Fred for the link to the Neweton Microscope, here is the UK web
store:
http://newtonmicroscopes.com/store/index.php?main_page=index&cPath=65

I hadn't heard of this project, and the price is great.

Ben

--
Research Support Manager
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271649
{http://www.mrc.ox.ac.uk/}

On 30/09/2013 21:04, "FMonson-at-wcupa.edu" {FMonson-at-wcupa.edu} wrote:

}
}
}
} --------------------------------------------------------------------------
} --
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America

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From noreply-at-ailaleadershipblog.com Tue Oct 1 10:08:19 2013
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Listers,

Below is the link provided to me by our student Brandon Saller linking you
to a PDF document citing the image shift issue seen on the EBSD scan

https://www.dropbox.com/sh/d7rzck8ufc4mi1k/ir7iYSEb3g

Thank you in advance for your response.

Fred Hayes
UC Davis

Dear Microscopy Community,

Recently we have purchased an EBSD detector for our Philips/FEI XL-30 SFEG
SEM in order to map nanocrystalline/ultra-fine grained materials. A problem
we are consistently running into is a shift in the map occurring
approximately every 40 minutes. This is most noticeable when attempting to
map the nanocrystalline grains with step sizes on the order of 10-50 nm.
Attached you will find a PDF documenting this issue and some of the steps we
have taken to resolve this issue, to no avail. We gratefully ask for your
help in diagnosing this issue, especially if you have run into a similar
issue before.

Thank you very much and best regards.

Brandon Saller
3rd year Materials Science Ph.D. student
University of California, Davis"

Fred Hayes
Manager
Advanced Materials Characterization and Testing Lab (AMCaT)
Dept of Chemical Engineering and Material Science
3001 Ghausi Hall
Univ of CA Davis
Davis, CA 95616
530-752-0284
chms.engineering.ucdavis.edu/index.html
chms.engineering.ucdavis.edu/research/amcat/index.html

==============================Original Headers==============================
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15, 22 -- Subject: link to PDF file for EBSD issue
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Email: hu.duan-at-averydennison.com
Name: Hu Duan

Organization: Avery Dennison

Title-Subject: [Filtered] Good condition Joel FE-SEM available for donation

Message: Due to recent lab consolidation, we have to retire a field emission SEM. The model is Joel
6330F, with Oxford INCA EDS. The instrument is 15 years old. But it is at excellent condition
because we had full service contract since day one. The instrument is at our California, Pasadena
facility. We would prefer to donate to an educational institute because of financial restrictions.
For inquiry, please contact me directly at hu.duan-at-averydennison.com. I am sure the instrument can
keep running as a great research or educational tool for you.

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Dear All, may i ask beside Ditabis which are selling the image plate and scanner for TEM , which other vendors are available in the market?

Cheers,
Yee Yan, Tay
FACTS Lab
MSE, NTU
Singapore

==============================Original Headers==============================
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Greetings,
As we are planning on a move to a new facility, we have been looking at
liability insurance coverage for our instruments during the move itself.
Although, vendor's insurance covers de-install/re-install, it does not
cover any instrumentation insurance during the transport from one location
to the other. The question has been raised if we would like to continue
insurance coverage (though our university) after installation in the new
facility.

Our question for list members is....do you have insurance on you major
instrumentation either through your facility or a private firm that would
cover major issues (flood,storm,fire etc) not covered with your standard
service contract? Also do you have suggestions of any insurance providers
specializing in instrumentation insurance during the move.

Thanks,
Amanda Lawrence
Institute for Imaging and Analytical Technologies
Mississippi State University

==============================Original Headers==============================
5, 22 -- From ALawrence-at-i2at.msstate.edu Wed Oct 2 07:53:45 2013
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Good morning, Amanda,

I've installed/moved/relocated 19 electron microscopes here (several microscopes were moved multiple times) at the University of South Florida (I have worked in 2 Departments, relocating labs in both Departments, as well as setting up a Core in another Department). We used national moving companies such as Allied Movers to do the work, using forklifts and pallet jacks to move the instruments from lab to lab, and building to building. We moved one TEM the length of 2 football fields on a forklift, down sidewalks. With moving companies, the company is bonded against loss. If they damage your instrument, they will be responsible for the damages. We always paid our microscope service providers for disassembly and reassembly, and had site surveys performed prior to installation to insure that the instruments would operate at specification when installed. We never purchased additional insurance, and do not carry additional insurance on our instruments. We keep service contracts on our microscopes. We have only (once) suffered one cosmetic scratch to one of the countertops on one of the TEM's.

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: ALawrence-at-i2at.msstate.edu [ALawrence-at-i2at.msstate.edu]
Sent: Wednesday, October 02, 2013 9:01 AM
To: Haller, Edward

Greetings,
As we are planning on a move to a new facility, we have been looking at
liability insurance coverage for our instruments during the move itself.
Although, vendor's insurance covers de-install/re-install, it does not
cover any instrumentation insurance during the transport from one location
to the other. The question has been raised if we would like to continue
insurance coverage (though our university) after installation in the new
facility.

Our question for list members is....do you have insurance on you major
instrumentation either through your facility or a private firm that would
cover major issues (flood,storm,fire etc) not covered with your standard
service contract? Also do you have suggestions of any insurance providers
specializing in instrumentation insurance during the move.

Thanks,
Amanda Lawrence
Institute for Imaging and Analytical Technologies
Mississippi State University

==============================Original Headers==============================
5, 22 -- From ALawrence-at-i2at.msstate.edu Wed Oct 2 07:53:45 2013
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12, 41 -- From: "Haller, Edward" {ehaller-at-health.usf.edu}
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12, 41 -- "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
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Anyone else notice the Zeiss EM-109 in the background of the Big Bang Theory tonight?

--Justin A. Kraft

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X-from: ravi.thakkar369-at-gmail.com ()

That orange column has guaranteed it a bit role in all science related movies and programs. I wonder if Zeiss had any idea of the impact of that orange column. A happy accident or crafty planning on their part to keep their product in the eye of the scientific community?

-----Original Message-----
X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
Sent: Thursday, October 03, 2013 8:33 PM
To: Frank Karl

Anyone else notice the Zeiss EM-109 in the background of the Big Bang Theory tonight?

--Justin A. Kraft

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Dear Frank,
Dear Listers,

My short comment here with regard to this "historic" thread might be misplaced.

The {Orange} ["Zeiss-RED"] - at least for TEM's will be missed (I hope so) by some TEM-ists:

ZEISS-TEM's will not be produced any more...(you certainly have heard or read about).

Anyone needs a ZEISS EM 109 / differential pumping system, + analog 35 mm camera option... vintage 1976,
in work since 02/1981, beloved (most of the time) and hated (for its sporadic bullishness and stubbornness when once failing)..
Functional, last (annual) service: Jan. 2013....?
Available at: EM-Lab, Pathology, SALK-LKH/PMU SALZBURG, Austria, at the latest July 2014 (;-((

Best regards,
have a nice and beautiful weekend,

Wolfgang MUSS

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} Gesendet: Freitag, 04. Oktober 2013 14:26
} An: Mu� Wolfgang
} Betreff: [Microscopy] RE: TEM sighting.
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} That orange column has guaranteed it a bit role in all science related
} movies and programs. I wonder if Zeiss had any idea of the impact of
} that orange column. A happy accident or crafty planning on their part
} to keep their product in the eye of the scientific community?
}
}
} -----Original Message-----
} X-from: kraftpiano-at-gmail.com [mailto:kraftpiano-at-gmail.com]
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} Subject: [Microscopy] TEM sighting.
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} Anyone else notice the Zeiss EM-109 in the background of the Big Bang
} Theory tonight?
}
} --Justin A. Kraft
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Hi, Ravi,

In a quick read on the subject, it seems that the primary diagnosis is made by immunohistochemical staining of skin biopsy sections for notch-3 around blood vessels in smooth muscle cells, not something done by electron microscopy. Immunohistochemistry would be much less expensive, and faster. Why would you want to employ E.M. as a diagnostic tool in this case? I am not currently running a clinical E.M. lab, but I ran one for 15 years. If you need E.M. to confirm your findings, you can chemically convert the DAB reaction produced in the smooth muscle cells labeling notch-3 to darken with osmium tetroxide to confirm that you are labeling smooth muscle cells, then run your samples up for TEM. Otherwise, you would need to look for granular osmiophilic deposits of notch-3 accumulating in the vascular smooth muscle cell cytoplasmic membranes by E.M. http://en.wikipedia.org/wiki/CADASIL Mutations in the Notch 3 gene (on the short arm of chromosome 19) cause an abnormal accumulation of Notch 3 at the cytoplasmic membrane of vascular smooth-muscle cells both in cerebral and extracerebral vessels,[4] seen as granular osmiophilic deposits on electron microscopy.[5]
[5] Ruchoux MM, Guerouaou D, Vandenhaute B, Pruvo JP, Vermersch P, Leys D (1995). "Systemic vascular smooth muscle cell impairment in cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy". Acta Neuropathol. 89 (6): 500�12. doi:10.1007/BF00571504. PMID 7676806.

Ed

Edward Haller, Lab Manager
University of South Florida
Integrative Biology Department
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
813-974-2676
ehaller-at-usf.edu
Office: ISA 1046
________________________________________
X-from: microscopylistserver-noreply-at-microscopy.com [microscopylistserver-noreply-at-microscopy.com]
Sent: Thursday, October 03, 2013 10:31 PM
To: Haller, Edward

X-from: ravi.thakkar369-at-gmail.com ()

We have been receiving skin biopsies for the last several years. We have the same problem finding CADASIL by TEM as well. My understanding is that it looks like deposits you see in a kidney biopsy and are found in the blood vessels of the dermis, am I wrong in this?

Does anyone know who came up with the idea that CADASIL could be seen by TEM?

Ron Austin
Dept. of Pathology
LSU Medical Center
Shreveport, LA

-----Original Message-----
X-from: microscopylistserver-noreply-at-microscopy.com [mailto:microscopylistserver-noreply-at-microscopy.com]
Sent: Thursday, October 03, 2013 9:26 PM
To: Austin, Ronald

X-from: ravi.thakkar369-at-gmail.com ()

All:
Try Goggle "CADASIL skin biopsies".

http://www.ncbi.nlm.nih.gov/pubmed/9228269 "Six CADASIL patients presented with symptoms ranging from migraine to severe tetraparesis with dementia. Two clinically unaffected patients had abnormal MRIs. On MRI 7 patients showed various degrees of leucoencephalopathy. One 22-year-old woman with migraine had a normal MRI. Granular, electron dense, osmiophilic material (GEM) was found in skin biopsies of all 8 patients including the 22-year-old woman with migraine and a normal MRI. As shown by genetic linkage analysis she was carrying the disease haplotype. GEM was not found in the control group."

http://www.ncbi.nlm.nih.gov/pubmed/20809020 "Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is an inherited vascular disorder, non-amyloid and non-atherosclerotic, affecting predominantly the central nervous system. We examined samples of skin biopsies from six patients (men, 43-52-year-old), admitted for treatment in the Neurology Clinic regarding the presence of partial motor impairment on upper and lower right limbs, facial asymmetry and phrasing impairment (three of the patients); These three patients had family history remarkable for early-onset strokes: mother and two brothers deceased by early strokes (40-50-year-old). Skin biopsy samples were fixed in glutaraldehyde and post-fixed in osmium tetroxyde. After dehydration, tissue samples were embedded in Epon. Ultrathin sections were mounted on copper grids and stained with uranyl acetate and lead citrate as usual and examined with a transmission electron microscope Phillips CM100. In all cases ultrastructural study showed granular osmiophilic material (GOM) in extracellular locations, between degenerating smooth muscle cells in dermal arteries or in their indentations. Deposits of GOM varied in size and electron density. Degeneration and loss of smooth muscle cells (SMCs) leads to abnormal enlargement of the space between these cells Ultrastructural analysis in three cases showed chromatin condensation and peripheral aggregation of nuclear material suggesting cells entry to apoptosis. These aspects and the marked destruction of the vascular wall were correlated with MRI findings and the severity of clinical manifestations at these patients. Our study showed that findings of GOM deposits, degeneration and loss of SMCs (probably by apoptosis), cell adhesion elements disturbance are characteristic for CADASIL disease and sufficient for diagnose of certainty. Moreover, electron microscopy analysis of skin biopsies is a useful tool for a differential diagnosis and can be considered as first choice m!
ethod."

http://www.neurology.org/content/59/6/961.full

John R Willis
Technical Writer
Valcor Engineering Corp
2 Lawrence Road
Springfield, NJ 07081
Voice (973) 467-8400 Ext 7257
FAX (973) 467-8382
johnwillis-at-valcor.com
www.valcor.com

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-----Original Message-----
X-from: microscopylistserver-noreply-at-microscopy.com [mailto:microscopylistserver-noreply-at-microscopy.com]
Sent: Thursday, October 03, 2013 10:29 PM
To: John Willis

X-from: ravi.thakkar369-at-gmail.com ()

Dear Listers,

apologize, if I will try to comment a little bit late on this topic from my own experience.

I am just trying to work up 3 CADASIL-specimens
(skin – deep dermal excision/punch biopsies) from inner upper arm) I got recently for confirming the suspicion of CADASIL.

Let me resume:
on Di/Tues 10.09.2013 01:51 we received a post on this by
dwolfe-at-capitalhealth.org Name: Dee Wolfe, who asked for help in finding an outside Lab for EM-Analysis ( {submit a skin biopsy to test for Cadasil (granular osmiophilic material, or GOM)} )
on Mi/Wed 11.09.2013 15:13 Dee Woolfe replied:
“Thanks to all who replied to my post regarding cadasil.
Who knew there would be so many options to choose from after my web search had come up empty?
I have forward your replies to our histology supervisor.
Now he can let our neuro people know we can get it done.”

Unfortunately he did not tell us at least a short summary of responses he had received, which would have been interesting (who / which labs, are doing what?) .

On Fr 04.10.2013 04:27 I found the post of ravi.thakkar369-at-gmail.com
“We are receiving 8-10 per year skin biopsy for CADASIL, but we are unfortunate to prove it by TEM. Recently we got one strongly suspected case with MRI features, but again we are unlucky to find it on TEM.”

And, following thereafter, the 3 replies
by Ed Haller (“primary diagnosis is made by immunohistochemical staining of skin biopsy sections for notch-3 around blood vessels in smooth muscle cells, not something done by electron microscopy.”),

by Ron Austin („We have been receiving skin biopsies for the last several years. We have the same problem finding CADASIL by TEM as well. My understanding is that it looks like deposits you see in a kidney biopsy and are found in the blood vessels of the dermis, am I wrong in this? Does anyone know who came up with the idea that CADASIL could be seen by TEM?”) and most recently

by John R Willis (try google, coming up with 3 URLS for interesting articles from PubMed: 1997; 2002 (NeuroImages) and most interesting an article from Romania, 2010, see also below in Willis's message).

OK.
With regard to the issue:

- EM for diagnosing CADASIL i) choice of EM in general, and
- EM for diagnosing CADASIL ii) by use of skin biopsies:

ad i) due to the various and multiple mutation variants, diagnostic (T)EM (not only because I have done that for the last 33 years) seems to be also these days the 1st choice since GOM’s by TEM always were found also in patients not exhibiting NOTCH3 mutations sought to be expressed using IHC or mutation analysis (as can be seen from various articles I have in files).
Genetic mutation analysis etc. therefore may be the “search for the needle in the haystack” and therefore be expensive and cumbersome.
cf: Diagnostic strategies in CADASIL, H.S. Markus, FRCP; R.J. Martin, MRCP; M.A. Simpson, BSc; Y.B. Dong, PhD; N. Ali, MSc; A.H. Crosby, PhD; and J.F. Powell, PhD in: NEUROLOGY 2002;59:1134–1138
} } ….Diagnosis by mutation detection can be a major undertaking in many patients because the notch3 gene is a large gene with 33 exons encoding for a protein of 2,321 amino acid residues. The extracellular portion of this protein contains 34 tandem repeats of an epidermal growth factor (EGF) motif, each of which contains six cysteine residues binding within the domain as three cysteine-cysteine disulphide bonds. The mutations that have been demonstrated in CADASIL occur in these EGF repeats, which are encoded for by the first 23 exons.4 Screening all of 23 exons is very time consuming and it is not realistic to offer it as a routine clinical service. It has been reported that 60 to 70% of mutations cluster in exons 3 and 4 and therefore a limited screen can be achieved by examining these exons alone.4 Nevertheless, this approach will miss approximately a quarter of cases. { {

There have been a lot of articles on the subject, not only the papers mentioned by my fore-posters, but also especially in Ultrastructural Pathology. But all authors state in “Methods”: “skin biopsies”, only one article states “skin biopsy from upper arm”

ad ii) from several references I conclude that - if present - not only skin vessels show these GOMs. There are papers reporting muscular tissue (intramuscular vessels) as well as nerve tissue (vessels in N. suralis) also should be taken into consideration. Invasive techniques always are questioned more and more so if there is -for some reasons- a muscle biopsy the patient will or will not accept having another biopsy from skin (in another location than for muscle biopsy)
cf: Cerebrovasc Dis 2007;24:401–404: Neuromuscular Implications in CADASIL, by Josef Finsterer

Within my 3 skin biopsies I am searching now on some deeper dermis levels within my ultrathin sections.
Until now I was not able - for only some sections - to find unequivocal structures resembling the so called GOMs.
It seems (at least to me) that one can find either GOMs at first sight (eventually accidentally - but fortunately) or has to look for them by examining multiple / further consecutive ultrathin sections:
cf:
SCHULTZ A., Santoianni R., Hewan-Lowe K., Stern B., Hunter S.
Vasculopathic Changes of CADASIL can be focal in Skin Biopsies.
Ultrastructural Pathology 23 / 4, 241-247, 1999 (s. Originalheft und Kopie in Diagnostik-Ordner)

ex Abstract:
{ {CADASIL is a newly described cause of vascular Dementia. Pathologic Examination shows multiple small infarcts in the deep cerebral white matter together with a nonatherosclerotic, nonamyloid angiopathy involving the media of small cerebral arteries.
Ultrastructurally, characteristic granular material is present in the basal lamina of vascular smooth muscle cells in cerebral and extracerebral blood vessels.
The ultrastructural changes have also been demonstrated in skin biopsies of affected patients; consequently, some investigators have recently recommended skin biopsies for the diagnosis of CADASIL.
This study describes a 54-year old male with a family history for strokes who had clinical and radiological features suggestive of CADASIL.
A skin biopsy was performed to confirm the diagnosis. Initially, the characteristic vasculopathic changes of CADASIL were not identified within small blood vessel walls.
However, multiple deeper sections in other areas showed electron dense material asscociated with vascular smooth muscle cells, characteristic of CADASIL.
Subsequent genetic testing demonstrated a single nucleotide substitution at position 659 on chromosome 19p13.1 causing an amino-acid change (Cys--} Phe), a finding indicative of CADASIL.
The involvement of Blood vessels within the dermis makes skin biopsy a useful adjunct in the diagnosis of CADASIL.
HOWEVER, as illustrated by this case, the FINDINGS may be FOCAL, requiring a THOUROUGH EVALUATION of the entire biopsy specimen.”} }

or, IMHO the most forwarding article with standards of prep & workup of biopsies including images (LM-semithin as well as EM-ultrathin sections):
Tikka et al, Congruence between NOTCH3 mutations and GOM in 131 CADASIL patients, Brain 2009: 132; 933–939:
{ { .... Either full thickness punch biopsies or small incision biopsies were taken (usually from the upper arm).... The diagnostic skin biopsy should include the border zone between deep dermis and upper subcutis, where small arterial vessels of correct size are located. Detection of GOM requires technically adequate biopsies and distinction of true GOM from fallacious deposits. If GOM is not found in the first vessel or biopsy, other vessels or additional biopsies should be examined. ..... } }

I greatly should acknowledge receiving opinion or practical experience of other MSA-Listers on the subject CADASIL diagnosis by TEM.
If anyone interested in some articles on the matter I should be happy to share (please send short request with permission to send pdfs…Copyright issues: pdfs sent only {for personal use and learning} )

Thanking you for your interest and time
have a beautiful weekend,

best regards,

Wolfgang

Wolfgang MUSS PhD
EM-Lab,
Univ. Inst. Pathol. SALK-LKH & PMU SALZBURG
AUSTRIA
==========================================================================
Von: johnwillis-at-valcor.com [mailto:johnwillis-at-valcor.com]
Gesendet: Freitag, 04. Oktober 2013 15:59
An: Muß Wolfgang
Betreff: [Microscopy] Re: Skin biopsy for CADASIL

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All:

Try Goggle "CADASIL skin biopsies".
---------------------------------------------------------------
http://www.ncbi.nlm.nih.gov/pubmed/9228269
"Six CADASIL patients presented with symptoms ranging from migraine to severe tetraparesis with dementia. Two clinically unaffected patients had abnormal MRIs. On MRI 7 patients showed various degrees of leucoencephalopathy. One 22-year-old woman with migraine had a normal MRI. Granular, electron dense, osmiophilic material (GEM) was found in skin biopsies of all 8 patients including the 22-year-old woman with migraine and a normal MRI. As shown by genetic linkage analysis she was carrying the disease haplotype. GEM was not found in the control group."

Acta Neurol Scand. 1997 Jun;95(6):351-7.
CADASIL: skin biopsy allows diagnosis in early stages.
Ebke M, Dichgans M, Bergmann M, Voelter HU, Rieger P, Gasser T, Schwendemann G.
Source: Department of Neurology, Zentralkrankenhaus Bremen-Ost, Bremen, Germany.
Abstract
OBJECTIVES: Our aim was to investigate the diagnostic impact of skin biopsies in CADASIL patients.
MATERIALS AND METHODS: Eight consenting CADASIL patients belonging to a German-Caucasian kindred were assessed clinically, genetically, by MRI and skin biopsy. Skin biopsy results were compared to 5 patients suffering from sporadic leucoencephalopathies (control group).
RESULTS: Six CADASIL patients presented with symptoms ranging from migraine to severe tetraparesis with dementia. Two clinically unaffected patients had abnormal MRIs. On MRI 7 patients showed various degrees of leucoencephalopathy. One 22-year-old woman with migraine had a normal MRI. Granular, electron dense, osmiophilic material (GEM) was found in skin biopsies of all 8 patients including the 22-year-old woman with migraine and a normal MRI. As shown by genetic linkage analysis she was carrying the disease haplotype. GEM was not found in the control group.
CONCLUSION: Our findings substantiate the impact of skin biopsies in defining the carrier status in CADASIL families.
PMID: 9228269 [PubMed - indexed for MEDLINE]

-------------------------------------------------------------
http://www.ncbi.nlm.nih.gov/pubmed/20809020
"Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is an inherited vascular disorder, non-amyloid and non-atherosclerotic, affecting predominantly the central nervous system. We examined samples of skin biopsies from six patients (men, 43-52-year-old), admitted for treatment in the Neurology Clinic regarding the presence of partial motor impairment on upper and lower right limbs, facial asymmetry and phrasing impairment (three of the patients); These three patients had family history remarkable for early-onset strokes: mother and two brothers deceased by early strokes (40-50-year-old). Skin biopsy samples were fixed in glutaraldehyde and post-fixed in osmium tetroxyde. After dehydration, tissue samples were embedded in Epon. Ultrathin sections were mounted on copper grids and stained with uranyl acetate and lead citrate as usual and examined with a transmission electron microscope Phillips CM100. In all cases ultrastructural study showed granular osmiophilic material (GOM) in extracellular locations, between degenerating smooth muscle cells in dermal arteries or in their indentations. Deposits of GOM varied in size and electron density. Degeneration and loss of smooth muscle cells (SMCs) leads to abnormal enlargement of the space between these cells Ultrastructural analysis in three cases showed chromatin condensation and peripheral aggregation of nuclear material suggesting cells entry to apoptosis. These aspects and the marked destruction of the vascular wall were correlated with MRI findings and the severity of clinical manifestations at these patients. Our study showed that findings of GOM deposits, degeneration and loss of SMCs (probably by apoptosis), cell adhesion elements disturbance are characteristic for CADASIL disease and sufficient for diagnose of certainty. Moreover, electron microscopy analysis of skin biopsies is a useful tool for a differential diagnosis and can be considered as first choice method."

Rom J Morphol Embryol. 2010;51(3):455-7.
Electron microscopy analysis of skin biopsies in CADASIL disease.
Cotrutz CE, Indrei A, Bădescu L, Dacălu C, Neamţu M, Dumitrescu GF, Stefanache F, Petreuş T.
Source: Department of Cell and Molecular Biology, Faculty of Medicine, Grigore T Popa University of Medicine and Pharmacy, Iassy, Romania. cotrutz-at-yahoo.com
Abstract
Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy (CADASIL) is an inherited vascular disorder, non-amyloid and non-atherosclerotic, affecting predominantly the central nervous system. We examined samples of skin biopsies from six patients (men, 43-52-year-old), admitted for treatment in the Neurology Clinic regarding the presence of partial motor impairment on upper and lower right limbs, facial asymmetry and phrasing impairment (three of the patients); These three patients had family history remarkable for early-onset strokes: mother and two brothers deceased by early strokes (40-50-year-old). Skin biopsy samples were fixed in glutaraldehyde and post-fixed in osmium tetroxyde. After dehydration, tissue samples were embedded in Epon. Ultrathin sections were mounted on copper grids and stained with uranyl acetate and lead citrate as usual and examined with a transmission electron microscope Phillips CM100. In all cases ultrastructural study showed granular osmiophilic material (GOM) in extracellular locations, between degenerating smooth muscle cells in dermal arteries or in their indentations. Deposits of GOM varied in size and electron density. Degeneration and loss of smooth muscle cells (SMCs) leads to abnormal enlargement of the space between these cells Ultrastructural analysis in three cases showed chromatin condensation and peripheral aggregation of nuclear material suggesting cells entry to apoptosis. These aspects and the marked destruction of the vascular wall were correlated with MRI findings and the severity of clinical manifestations at these patients. Our study showed that findings of GOM deposits, degeneration and loss of SMCs (probably by apoptosis), cell adhesion elements disturbance are characteristic for CADASIL disease and sufficient for diagnose of certainty. Moreover, electron microscopy analysis of skin biopsies is a useful tool for a differential diagnosis and can be considered as first choice method.
PMID: 20809020 [PubMed - indexed for MEDLINE] Free full text
F Our study shows that skin biopsy can be used as confirmation diagnosis for CADASIL disease while genetic tests are more expensive and demanding. Even if irrelevant symptoms and minimal MRI changes are present, the electron microscopic examination shows at least GOM presence. Ruchoux MM et al. [9] were the first who describe the presence of GOM within the vessel walls of skin biopsies in a patient affected by this Electron microscopy analysis of skin biopsies in CADASIL disease
457
disease. Up to now, all skin biopsies in subjects carrying the CADASIL haplotype (100%) have been characterized by the presence of GOM [11]. These GOM are present in the SMCs basal lamina of the skin vessels or between them and appears to be unique and pathognomonic for CADASIL [11]
Artikel ist zumindest in FACTS(Q-Z)\SKIN\BIOPS's,-Y,techn als
{CADASIL_EM-ANAL of skin biopsies in CADASIL disease,&!EM+Abb. Cotrutz et al, Rom J Morphol Embryol. 2010(13-10-04pdf).pdf}
abgespeichert.

-------------------------------------------------------------------------------
http://www.neurology.org/content/59/6/961.full

DOI 10.1212/WNL.59.6.961
Neurology 2002;59;961
Skin biopsy findings in CADASIL
Brian W. Smith, Jean Henneberry and Timothy Connolly

Neuro Images
Skin biopsy findings in CADASIL
Brian W. Smith, MD, Jean Henneberry, MD, Timothy Connolly, BS, Springfield, MA
A 48-year-old woman was seen on neurology consultation at Baystate Medical Center for gait instability and abnormal CT scan of the brain. She had a medical history of stroke 5 years prior consisting of left-sided weakness with no residual symptoms. Her history also included seizures in the remote past, migraine without aura, and depression.
Neurologic examination was notable for evidence of cognitive impairment, dysarthric speech, and brisk reflexes.
MRI of the brain was obtained. There was diffuse white matter abnormality consisting of hyperintensity on T2 and fluid-attenuated inversion-recovery (FLAIR) imaging and cystic changes (figure A). The findings were suggestive of cerebral autosomal-dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL).1 A skin biopsy showed the characteristic granular deposits in the basal lamina of a small blood vessel on electron microscopy (figure B).2 She was later found to have a Notch3 gene mutation.

1. Davous P. CADASIL: a review with proposed diagnostic criteria. European Journal of Neurology 1998;5:219-233.
2. Mayer M, Straube A, Bruening R, et al. Muscle and skin biopsies are a sensitive diagnostic tool in the diagnosis of CADASIL. J Neurol 1999;
246:526-532. Address correspondence and reprint requests to Dr. Brian W. Smith, Bay State Medical Center, Department of Neurology, 759 Chestnut St., Springfield,
MA 01199-1001.

Figure. (A) FLAIR MR image showing extensive white matter signal hyperintensities in temporopolar regions and cystic changes. (B) Electron microscopy of skin biopsy shows electron dense granular deposits (arrows) in the basal lamina surrounding vascular smooth muscle cells.
Copyright © 2002 by AAN Enterprises, Inc. 961

Artikel ist zumindest in FACTS(Q-Z)\SKIN\BIOPS's,-Y,techn als
{CADASIL_Skin biopsy findings in CADASIL,&!EM+Abb. Smith et al, Neurol.2002(961.ful)l(13-10-04pdf).pdf}
abgespeichert.

=======================================
John R Willis
Technical Writer
Valcor Engineering Corp
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FAX (973) 467-8382
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Sent: Thursday, October 03, 2013 10:29 PM
To: John Willis

HI List

We find ourselves in the fortunate sitiation of being able to buy a
new EDX system for our TEM. It is STEM, old but functional good count
rate. Currently we have an Oxford Instruments INCA system (SiLi) and
although it does the job it does not tick all the boxes. We have been
told by the reps we have asked (Bruker, Thermo, Oxford) that SDD is
the way to go. I was wondering what the general opinion was out there
on the current systems available. I really have no experience on
systems other than Oxford Instruments so I would welcome coments and
suggestions about the relative merits (or otherwise) of the current
players. I would be interested to hear from others from outside the UK
as here as it is an oxford stronghold and I dont know many people who
have alternatives here in the UK. We are looking at fully
functionality as we have a broad userbase.

Thanks in advance

John

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Listers,
Does anyone have/know of an optical projection tomography setup? Ideally not too far from Purdue, Indiana but other locations would be of interest. I am happy to receive information from vendors.

Regards

Chris

Christopher J. Gilpin Ph.D.
Director, Life Science Microscopy Facility
Purdue University
Whistler Hall of Agriculture Research, Room S052
170 S. University St
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765-494-7750
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Greetings,

I am in the process of refurbishing an Amray AMR-1000 SEM. I would
like to remove the anode and column liner tube in order to inspect the
apertures. I do not have the special tools for doing this referred to
in the service manual so I was wondering:

1. Does anyone have a picture of these special tools? This would be
helpful in the case I fabricate a tool myself.

2. Is anyone familiar with workarounds for removing these parts
without the special tools?

Best,
Dr. Mark Bradshaw
Atkinson Thin Film Systems Inc.

==============================Original Headers==============================
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Hi, All-

I am trying to put a 1984 Balzers 400T freeze-fracture device back into
service, primarily for rotary shadowing. I totally do not remember at all
how the guns were wired in the chamber. If we had had smart phones in
those days, I'd have a picture, but I don't have any images nor a diagram.
A mental image just isn't coming to me. Does anyone have a diagram,
picture, or description?

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*
****************************************************************************

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Post-Doctoral Research Associate, Materials Science & Engineering-13000000YJ

Description

POST-DOCTORAL RESEARCH ASSOCIATE-Materials Science and Engineering- Pay
Grade , DOE&Q,We are looking to hire a highly motivated individual to
fill a post-doctoral position to work jointly at The University of
Tennessee, Knoxville and Oak Ridge National Lab. This position is to
obtain an atomic-level understanding of issues concerning multiferroic
materials through the combination of atomic-resolution electron
microscopy and electron energy loss spectroscopy using an
aberration-corrected scanning transmission electron microscope located
at ORNL. The research is to be performed at ORNL under the supervision
of Prof. Ramesh Ramamoorthy, UT Governor’s Chair and Deputy Director of
ORNL, Prof. Stephen J. Pennycook, leader of the ORNL STEM group and
currently an ORNL-UT joint faculty member, and Albina Borisevich, ORNL
staff member. The work will complement ongoing atomic-resolution
electron microscopy studies of ferroelectric materials in the Materials
Science and Technology Division at ORNL.

Topics to be addressed will primarily be the investigation of novel
materials for advanced energy applications. Studies will include
aberration-corrected electron microscopy of defects correlated with
electron energy loss spectroscopy and mapping of ferroelectric
displacements at high spatial resolution.

Qualifications

Candidates must have a PhD in materials science and engineering or
physics or similar discipline, with previous experience in
aberration-corrected electron microscopy, and a demonstrated track
record of publication in the scientific literature. Previous experience
with multiferroics and functional oxides is highly desirable.

Job

Research Professional

Primary Location

US-Tennessee-knoxville

Organization

Materials Science & Engr

Schedule

Full-time

Job Posting

Sep 17, 2013, 3:17:23 PM
--
Signature --
Albina Y. Borisevich
R&D Staff
Electron Microscopy Group
Oak Ridge National Laboratory
Materials Science and Technology Division
PO Box 2008
Oak Ridge TN 37831-6031

http://stem.ornl.gov/

For express mail add: 1 Bethel Valley Road
phone: (865) 576-4060
fax: (865) 574-4143

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Hiya all on the Microscopy List,

I was wondering if any of you fine people on this list could be of assistance.

I have had an unusual career path in that I moved from doing computer simulation research in physics and materials science and then worked in the computer games industry for a few years.

I'm now back in academia and looking to combine these together, and use computer graphics techniques to carry out simulations. I've had some previous success with this:
http://www.sciencedirect.com/science/article/pii/S1476927109000486

I'm interested in looking at using the idea of how lighting is simulated in simply graphics to model the production of SEM and TEM images.

So a few questions:
Is this something that would be of interest?
Is this something that someone has already tired?
Could anyone point me to an Electron Microscopy 101 text-book / web-page that outlines the physics/maths behind the production of these sort of microscopy images?

I should note that my PhD is in Theoretical Physics so I don't scare too easy on the maths front.

THANKS TO YOU ALL

SIMON

DR SIMON SCARLE rm 2Q20
-at-SimonScarle -at-UWEGames
Acting Program Leader & Senior Lecturer in Games Technology
Computer Science & Creative Technologies
University of the West of England
Frenchay Campus, Coldharbour Lane
Bristol, BS16 1QY
simon.scarle-at-uwe.ac.uk
Tel: 01173 285 512

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Email: Amy.Koshoffer-at-UC.Edu
Name: Amy Koshoffer

Organization: University of Cincinnati

Title-Subject: [Filtered] repair for RMC MT6000-XL

Message: I am looking for someone who can repair our ultramicrotome. It is not automatically
advancing anymore when we are thin sectioning.

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Email: rjpe-at-cellbio.duke.edu
Name: Robert Perz-Edwards

Organization: Duke University

Title-Subject: [Filtered] TEM position open at Duke University

Message: Are you looking for a job in EM, but also interested in learning more? We're
hoping you'll join our team in the lab of Dr. Michael Reedy, at Duke University, in
Durham NC. We study the in situ mechanism of force generation by myosin
motors and the regulation of myosin by tropomyosin-troponin, using the
exquisitely well-ordered flight muscles of the giant waterbug, Lethocerus. Were
striving for a 1-ms, 1-nm resolution movie of how muscle works by integrating
time-resolved cryo-EM with real-time muscle physiology monitored by live
action X-ray diffraction movies.

Job opening is flexible and we will consider any level candidate from bachelor's
degree and up, including post-docs or research scientists. Exact job title and
pay level will depend on your experience. Current funding is guaranteed until
2017-2018.

Ideally, you are well versed in routine biological TEM, including fixation,
embedding, thin sectioning, and negative stain. You should also have
experience in, or willingness and ability to learn, 3D cryo-EM and electron
tomography, including single particle methods, iterative helical real-space
reconstructions and sub-volume averaging. EM will be most (~75%) of your
duties, but you will also assist with muscle physiology and X-ray diffraction
experiments as needed.

Please contact Robert Perz-Edwards (hiring manager), and send me your CV,
including contract information for 2-3 references.

Robert J Perz-Edwards, PhD
Assistant Research Professor, Reedy Lab
Cell Biology, Duke University
Box 3011 (US Mail)
303 Research Drive, Sands Bldg, Rm 457
Durham, NC 27710
919-684-5674
rjpe-at-cellbio.duke.edu

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Realname - Shakira O'Neil
Email - sjoneil05-at-gmail.com
ORGANIZATION - University of Pittsburgh
EDUCATION - Graduate College
SUBJECT_OF_QUESTION - Possibly Shopping for Inverted Microscope
QUESTION - I am interested in shopping for an inverted microscope with a
chamber, controller, and stage. We would like to use this instrument for
imaging cell movement and gap junction protein dynamics. I am not
interested in receiving quotes from vendors just yet. I was wondering
what would be the best supplier and microscope to go with. My boss gave
the figure of 10K which I know is a long shot, but does anyone have any
ideas? Thank you

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Anyone know of a webcam that has this feature?? I'm still looking for multiple ways to replace 4X5 Polaroid.

Jeff Day, BA-LHC (Physics)
Tx ii/EM Analyst-20+ yrs./Training/Project Modifications
wa5ekh-at-juno.com
cell: 972537-7660(direct/text/voicemail)

____________________________________________________________
Fast, Secure, NetZero 4G Mobile Broadband. Try it.
http://www.netzero.net/?refcd=NZINTISP0512T4GOUT2

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***************************************************************************************
Forwarded from "Ask a Microscopist"
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****************************************************************************************

realname - Seongjin Jang
Email - jangseongjin-at-gmail.com
ORGANIZATION - Stevens Tech
EDUCATION - Graduate College
LOCATION - Hoboken
SUBJECT_OF_QUESTION - Bake out service for FEI Quanta FEG 450
QUESTION -
I need to bake out service due to the vacuum problem at 30kV.
Can you let me know any service provider except FEI for that service?
Can you let me know if I can do it by myself?
Then, please help me how to do it.
I appreciate your help.

==============================Original Headers==============================
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Email: pedromfjcosta-at-gmail.com
Name: Pedro Costa

Organization: KAUST

Title-Subject: [Filtered] TEM post-doctoral position available

Message: One Postdoctoral Research Fellow position is available immediately at the Physical Sciences
and Engineering Division of the King Abdullah University of Science and Technology (KAUST). The
position is particularly well suited for an electron microscopist with interests in both in situ
methods and carbon nanostructures. A PhD degree, within 5 years of receipt, in physics, materials
science, chemistry or related areas is mandatory. Experience in one or more of the following
sub-areas in TEM is necessary: in situ probing, high resolution imaging, EELS and FIB sample
preparation/manipulation. More details can be found at the employment section of
http://www.kaust.edu.sa (requisition 2745BR).

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Email: erwright73-at-gmail.com
Name: Elizabeth Wright

Organization: Emory University

Title-Subject: [Filtered] Postdoctoral Fellow Position, Emory University

Message: NIH-funded positions are available in the Wright laboratory at Emory University starting as
soon as possible (http://electronmicroscopy.emory.edu). Research in the lab is focused on bacterial
and viral structural biology, with emphasis on 3-D structure/function relationships between host
cells and viruses. Areas of interest include cryo-electron tomography of bacteria, bacteria �
bacteriophage interactions, paramyxoviruses, and HIV-1. Technology development interests are in the
areas of correlative microscopy and phase plate cryo-microscopy.
Candidates should have an interest in structural biology, bacteriology or virology, and should enjoy
working as part of an active research team. Background experience in one (or more) of the following
fields is desirable: bacteriology, virology, (cryo-) electron microscopy, and/or image
processing/analysis techniques.

The wet laboratory is within the Division of Pediatric Infectious Diseases and is fully equipped for
all aspects of microbiology, molecular biology, molecular virology, and protein biochemistry. The
Wright lab has complete access to a JEOL JEM-2200FS 200 kV FEG TEM with an in-column energy filter,
Zernike-style phase plates, and high-resolution 4kx4k CCD camera; a JEOL 1400 120 kV TEM with 2kx2k
camera; an FEI Vitrobot and a manual grid plunge freezer; a Bal-Tec high pressure freezer and Leica
freeze substitution system; a Leica cryo-ultramicrotome; an Improvision spinning disk confocal
microscope; and a Zeiss TIRF microscope. Emory is a vibrant, multidisciplinary campus with strong
ties to Georgia Tech and other universities within Georgia. Access to facilities in NMR, Mass
Spectrometry, and X-ray crystallography are available.

Applicants should include a cover letter describing research experience and interests, curriculum
vitae, and names and contact information for three references.

Dr. Elizabeth R. Wright
Emory University
Department of Pediatrics
2015 Uppergate Drive, NE Room: 548
Atlanta, GA 30322
erwrigh(at)emory.edu

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Need Surplus EDS system for JEOL T330A -Detector not necessarily required to be operational-available for donation to very low budget College start-up EM Lab.

Jeff Day
Texas

____________________________________________________________
Do THIS before eating carbs (every time)
1 EASY tip to increase fat-burning, lower blood sugar & decrease fat storage
http://thirdpartyoffers.juno.com/TGL3131/526000f838bf7f81b9est04vuc

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Colleagues,
Please bring this opportunity to the attention of anyone you
know who might be interested. Thanks. TB

A postdoctoral position is available to study the biophysics of
cellulose synthesis. The project aims to apply high resolution and
single-molecule sensitive imaging techniques to characterize the
motility of the cellulose synthase. As material we will use intact
plants (established lines and new ones to be developed) and we are
particularly interested in using in vitro preparations to achieve
greater opportunities for experimental manipulation. The position
requires a Ph.D. in a relevant area of biology or biophysics and
preference will be given to candidates having experience with
purifying active protein complexes. The position is supported by an
NSF grant to Lori Goldner (Physics) and Tobias Baskin (Biology). The
Goldner lab uses single-molecule-sensitive physical and optical tools
to study the dynamics and interactions of biomolecules and
bio-molecular complexes. The Baskin lab studies the role of cellulose
in growth. The University of Massachusetts Amherst is an Equal
Opportunity Employer and we encourage all qualified persons to apply.
To apply, please send your cv to baskin-at-bio.umass.edu. More
information about our respective labs can be found at
http://people.umass.edu/lgoldner and
http://www.bio.umass.edu/biology/baskin. Please feel free to contact
either of us for more information.

==============================Original Headers==============================
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Email: bidinozor-at-gmail.com
Name: bidin ozor

Organization: molecular biologist

Title-Subject: [Filtered] undesired photo from fluorescence microscope

Message: I did not stain my cells which are MDA-MB-231 cells; however I see something like a
specific molecule and especially bright nucleus under blue filter. I am lost.. Is there any
explanation? click to link to sample photograph
https://fbcdn-sphotos-h-a.akamaihd.net/hphotos-ak-prn2/v/1394859_10151952789952220_1564603189_n.jpg?oh=4ba5abd50dc37ae437756846f9f95d23&oe=5260D257&__gda__=1382120997_fe90216cbe0aaf81df10c4e9187cd612

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Email: scott.walck.ctr-at-mail.mil
Name: Scott Walck

Organization: Army Research Lab

Title-Subject: [Filtered] FFT and Scherzer focus question, Au on Ge test sample

Message: I've got a question about using the FFT for focusing a high magnification image in the FETEM.

When you are observing the rings of an amorphouse materials in the FFT, they move outward to a max
and then come back in as you go through focus. I thought that when the zero of the inner most ring
reaches it's max, that focus corresponds to the first zero in the contrast transfer function where
it is a max in 1/d space. This does correspond to Scherzer defocus condition where you are getting
the maximum information through the lens, correct? Or does this correspond to the minimum contrast
condition? The reason that I'm asking is that it appears that it corresponds to the minimum
contrast condition and I know that Scherzer should be at an underfocused value from the minimum
contrast focus.

Does anyone know where I might be able to find a gold particle on Ge film test sample?

Thanks in advance.

-Scott

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Email: hlowers-at-usgs.gov
Name: Heather Lowers

Organization: U.S. Geological Survey

Title-Subject: [Filtered] Colorado Microbeam Meeting

Message: Hello Everyone

The Mountain States Society of Electron Microscopists and the Colorado Microbeam Analysis Society
announce their Fall Dinner Meeting.

Paul Carpenter from Washington University in St. Louis will present "Practical Guidelines for
Microanalysis Using EDS and WDS". Come for the talk and networking opportunities with vendors and
folks in the Front Range area who specialize in microscopy and microanalysis in the fields of
biology, material science, geology, chemistry.

This year the meeting will be held on Wednesday October 30 from 6:30-9P at the Wynkoop Brewery in
downtown Denver on the heels of the Geological Society of America meeting. A buffet dinner will be
served. Costumes optional!

The cost is $35 for professionals and $20 for students. Nonmembers are welcome to attend.

Please RSVP by 10.22.2013 to hlowers678-at-hotmail.com

See attached flyer for details. Please pass along to those you feel would be interested.

Look forward to seeing you at the meeting!

http://comas.geoloweb.ch/index.php

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Email: modla-at-dbi.udel.edu
Name: Shannon Modla

Organization: University of Delaware

Title-Subject: [Filtered] Glow Discharge Problems

Message: Hi Everyone,

Our lab has a Peco EasiGlow glow discharge unit. Relatively recently the unit was having difficulty
achieving the required vacuum for performing glow discharge. We discovered that the pirani gauge
may have become contaminated with oil, so we ordered a new gauge and replaced it. Ever since, the
unit appears to be working normally (purple glow and all that).

However, since the fix, we've had some sporadic problems when glow discharging grids for negative
staining. In some instances (not every time) when examining glow discharged grids that have been
negative stained with uranyl acetate or PTA, we notice a very poor staining of the sample that
appears as though the stain precipitated (small dark blobs all over). Moreover, the grids cause a
distortion of the electron beam, leading to horrific astigmatism. We've run samples side-by-side
with and without the glow discharge. Grids not glow discharged look fine.

I was wondering if anyone may have any idea about what is going on. Are the grids (we primarily use
copper) somehow becoming magnetized or too highly charged by glow discharge? We use the default
Auto Run parameters on the glow discharge unit (negative glow polarity, 0.39 mBar vacuum set point,
15 mA discharge current set point, 60 second glow time). I've tried reducing the glow time by half,
but we still see the problem. Once again, the issue doesn't happen every time we use the unit but
enough that I'm starting not to trust it--so I end up running two sets of grids per sample (with and
without glow discharge) or just skip the glow discharge all together.

If anyone has any ideas on how to fix the problem, I'm all ears!

Thanks,
Shannon

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Dear Bidin Ozor,

The threadlike structures look like mitochondria to me.

Did you add a mounting medium containing DAPI? If so, it could be staining mitochondrial DNA. Usually nuclei are much brighter than mitochondria with DAPI, but I'll speculate that the DAPI is at such a high concentration that it is self-quenching in the nucleus.

I'd be happy to hear other ideas though.

Hope this helps,
Theresa

------------------------------------
Theresa C. Swayne, Ph.D.
Manager, Confocal and Specialized Microscopy Shared Resource
Herbert Irving Comprehensive Cancer Center, Columbia University
1130 Saint Nicholas Ave, 222A
New York, NY 10032

212-851-4613

tcs6-at-columbia.edu
http://hiccc.columbia.edu/research/sharedresources/confocal

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Dear All,

The Laboratory of Cellular Imaging and Macromolecular Biophysics is
seeking a motivated and creative postdoctoral researcher to develop new
approaches for determining the three-dimensional structure of cells based
on scanning transmission electron tomography, serial block-face scanning
electron microscopy, and related techniques.

This position is in the intramural program of the National Institute of
Biomedical Imaging and Bioengineering (NIBIB), which is part of the
National Institutes of Health, and is located in Bethesda, Maryland. Our
research group develops new methods in electron microscopy. For example,
we have previously developed techniques based on STEM, EELS and electron
tomography for application to biological systems. Current biological
interests of the group include the ultrastructure of stem cells, endocrine
pancreatic cells, and neurons.

Suitably qualified candidates should have a Ph.D. in the physical sciences
(physics, materials science) or in biophysics. Applicants should have
superior communication skills in English and a demonstrated record of
accomplishment. NIBIB is committed to integrating engineering and physical
sciences with the life sciences to advance basic research. Training in biology
is not essential for this position but there will be ample opportunities to
collaborate with and learn from world-class biologists. Preference will be
given to candidates with less than three years of postdoctoral experience.

NIH offers excellent salary and health care packages to its trainees.

The NIH is dedicated to building a diverse community in its training and
employment programs.

See also NIH web page at:

https://www.training.nih.gov/postdoc_jobs_nih/view/_31/2162/Electron_Microscopy_and_Tomography

Please send applications to:
Dr. Richard Leapman
Laboratory of Cellular Imaging and Macromolecular Biophysics
National Institute of Biomedical Imaging and Bioengineering
National Institutes of Health
Bldg. 13, Rm. 3N17
13 South Drive
Bethesda, MD 20892
Tel: 301-496-2599
E-mail: leapmanr-at-mail.nih.gov
Web page: http://irp.nih.gov/pi/richard-leapman

-------------

Please do not hesitate to contact us for further information.

Best regards,

Richard Leapman

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Hi Scott,

the bright rings of the FFT give positive contrast while the dark rings
negative contrast. I think you are right about the Scherzer defocus
giving the best resolution without pesky contrast reversals built into
the image; and the difference between Scherzer and "Gaussian focus"
(where you get minimum contrast) is not that large (a few clicks).

Gaussian is where the Fresnel fringes go away along the edge of any
holes in the sample (defocus = zero). Scherzer is where the phase
change due to spherical aberration is cancelled out by that due to
underfocus, giving you the best practical resolution (see
http://www.ceos-gmbh.de/FAQ/informationlimit.html). With higher defocus
values it is possible to create a passband in the contrast transfer
function that will "pick-out" higher spatial frequencies but the
interpretation is going to be more work involving simulations. The
so-called information limit is more down to a "damping envelope" that
all the FFT rings have to stay within and your contrast just fizzles out.

I'm sorry but I don't know where you can buy Au on Ge, could you try to
make one?

Good luck
Rob

On 10/18/2013 8:56 AM, microscopylistserver-noreply-at-microscopy.com wrote:
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} Name: Scott Walck
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} Organization: Army Research Lab
}
} Title-Subject: [Filtered] FFT and Scherzer focus question, Au on Ge test sample
}
} Message: I've got a question about using the FFT for focusing a high magnification image in the FETEM.
}
} When you are observing the rings of an amorphouse materials in the FFT, they move outward to a max
} and then come back in as you go through focus. I thought that when the zero of the inner most ring
} reaches it's max, that focus corresponds to the first zero in the contrast transfer function where
} it is a max in 1/d space. This does correspond to Scherzer defocus condition where you are getting
} the maximum information through the lens, correct? Or does this correspond to the minimum contrast
} condition? The reason that I'm asking is that it appears that it corresponds to the minimum
} contrast condition and I know that Scherzer should be at an underfocused value from the minimum
} contrast focus.
}
} Does anyone know where I might be able to find a gold particle on Ge film test sample?
}
} Thanks in advance.
}
} -Scott
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I looking for a copy of a Sorval JB4 Manually operated Rotary Microtome operation and service manual, service notes or advice.

Jeff/N. Texas
wa5ekh-at-juno.com
cell: 972-537-7660(direct/text/voice mail)
____________________________________________________________
Do THIS before eating carbs (every time)
1 EASY tip to increase fat-burning, lower blood sugar & decrease fat storage
http://thirdpartyoffers.juno.com/TGL3131/52657d0edf1a57d0e1545st01vuc

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Organization: Spectra Technology Corp

Title-Subject: [Filtered] ETEC Autoscan Schematic

Message: Hello,
I'm restoring a ETEC Autoscan SEM for a local high school.
Does any know of a resource to get a schematic or manual for this machine.
thanks!
Jay

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Jay,
Send me your snail mail address and I'll send a DVD with User's Manual,
Service Manual, Diagnostic Manual, Schematics and some other misc. stuff.

If you want an 11" x 17" schematics book, I'll have to charge for it.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

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Title-Subject: [Filtered] ETEC Autoscan Schematic

Message: Hello,
I'm restoring a ETEC Autoscan SEM for a local high school.
Does any know of a resource to get a schematic or manual for this machine.
thanks!
Jay

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Dear all,

The Southern California Society for Microscopy and Microanalysis (SCSMM)
announce their Fall Meeting.

The meeting will be held on Thursday, November 7th from 5:30 pm at the CEM
Building (Room 101B) at the University of Southern California (USC). A
buffet dinner will be served.

Our fall meeting will be dedicated to the UCLA California NanoSystem
Institute (CNSI). The featured speakers will be Dr. Paul Weiss, Director
of CNSI, Fred Kavli Chair in NanoSystem Sciences, Distinguished Professor
of Chemistry and Biochemistry & Materials Science and Engineering at UCLA,
and Dr. Adam Stieg, Scientific Director of the Nano & Pico
Characterization Laboratory at CNSI.

In order to register, please RSVP by email at micromark-at-juno.com. Respond
no later than 5:00 p.m. Monday, November 4th, 2013.

The cost is $25 for professionals and $10 for students. Membership dues
can be paid at the door on November 7, 2013.

Nonmembers are welcome to attend.

Please pass along to those you feel would be interested.

We look forward to seeing you at the meeting!

http://www.scsmm.org/

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This year the Appalachian Regional Microscopy Society (AReMS) is joining
the local MRS, AVS and ASM societies for a 2013 Fall Meeting taking place
Nov. 14-15.

The meeting is to be held in the NCSU Hunt Library on Centennial Campus in
Raleigh, NC.

Nov. 14:

FREE Tutorials on SEM/EBIC, SEM/CL & Cryo-Ultramicrotomy and TEM Prep
(limited number of seats for all tutorials, $50 fee for TEM Prep tutorial)

Table Top SEM Community Outreach Event

Nov. 15:

Student Poster Session & Platform Presentations

AReMS Invited Speakers Include: MSA Tour Speaker Dr. Sarah Miller (Duke
University) & AReMS Young Investigator Award Recipient Dr. Dave Cullen
(ORNL)

To register for the meeting, find out more details about the events
planned, learn about special hotel rates and get directions to the meeting
venue please visit:

http://www.aif.ncsu.edu/mrs_asm_avs_arems/

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Hi,

I am looking for software to control different Gatan controllers :

* Smartset 900
* Smartset 901
* ITC 503

I am looking especially for labview vi.

Can you help me ?

Regards

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We are running an older Oxford Tetra(tm) backscattered electron system on our older Hitachi S-2460N SEM. The SEM has been a trooper through the years, but the Tetra BSE system has recently developed some nasty noise in the signal.

It is scan rate dependent and seems to track back to 60 Hz. If we enable the Sync-to-AC feature we get nice vertical lines on our screen. When we turn it off, we get a splattering of bright dots across the captured image. It appears to be 8.3 ms between lines which corresponds to 120 Hz or every zero crossing of the AC line.

Does anyone have some spare Tetra pieces around that they are willing to part with? Our system has the external box (PN 1108-091) that is controlled by a serial connection from the EDS computer. I suspect the problem is in that box, but it might be in the detector assembly/pre-amp.

I have posted pictures of our unit and some problem images on our lab server at ftp://www.marl.iastate.edu/equipment/tetra.

Looking again to this helpful community,
Warren Straszheim
Assoc. Scientist
Iowa State University
515-294-8187

==============================Original Headers==============================
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6, 33 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
6, 33 -- Subject: Need Tetra control/amp box
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Hi Warren,

I do not have the Tetra control box, but the 120Hz noise in sync with
the line frequency is telltale of either dried up capacitor (which is
the most likely, as it is a very common failure mode for aging
electronics) or (less likely) partially-failed diode in the bridge
rectifier of the power supply of your control box.

If you have around electronics tech with decent component-level
troubleshooting skills, he/she should be able to track this noise down
to one of power supplies and repair. Usually couple of capacitors from
local Radioshack is all it takes.... apart of a few hours of work, of
cause....

Valery Ray
=================================
PBS&T, MEO Engineering Co., Inc.
290 Broadway, Suite 298
Methuen, MA 01844 USA
Phone: +1-978-296-5063 - leave a message with call-back number
US Mobile: +1-978-305-0479
Skype: pbstmeo
E-mail: vray-at-partbeamsystech.com
Web: www.partbeamsystech.com
Web: www.freudlabs.com

On 10/23/2013 5:53 PM, wesaia-at-iastate.edu wrote:
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} We are running an older Oxford Tetra(tm) backscattered electron system on our older Hitachi S-2460N SEM. The SEM has been a trooper through the years, but the Tetra BSE system has recently developed some nasty noise in the signal.
}
} It is scan rate dependent and seems to track back to 60 Hz. If we enable the Sync-to-AC feature we get nice vertical lines on our screen. When we turn it off, we get a splattering of bright dots across the captured image. It appears to be 8.3 ms between lines which corresponds to 120 Hz or every zero crossing of the AC line.
}
} Does anyone have some spare Tetra pieces around that they are willing to part with? Our system has the external box (PN 1108-091) that is controlled by a serial connection from the EDS computer. I suspect the problem is in that box, but it might be in the detector assembly/pre-amp.
}
} I have posted pictures of our unit and some problem images on our lab server at ftp://www.marl.iastate.edu/equipment/tetra.
}
} Looking again to this helpful community,
} Warren Straszheim
} Assoc. Scientist
} Iowa State University
} 515-294-8187
}
}
} ==============================Original Headers==============================
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} 6, 33 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
} 6, 33 -- Subject: Need Tetra control/amp box
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Email: lamiller-at-illinois.edu
Name: Lou Ann Miller

Organization: Materials Research Laboratory, University of Illinois

Title-Subject: [Filtered] Job opening for an experienced Electron Microscopist

Message: Research Scientist(s) position
Please see the link below for the job information and to apply electronically online:

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*Note for those unfamiliar with University jobs here, a common question: Is the job for only one
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All academic positions at the University are appointed on a 9- or 12-month service basis, eligible
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{ { { { { { { { { { { { { { { { { {} } } } } } } } } } } }
Lou Ann Miller
Biological Electron Microscopy
Frederick Seitz Materials Research Laboratory
Lab Room 125 Office 374 MRL
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Urbana, IL 61801
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Email: robert-at-appfive.com
Name: Robert Stroud

Organization: AppFive

Title-Subject: [Filtered] Nov 8 - UCLA Electron Diffraction Workshop

Message: Workshop on Precession Electron Diffraction, and
Orientation & Strain Mapping in TEM

The UCLA Department of Material Science will be holding a workshop on November 8th, in Room 2101,
Engineering 5. from 9:00 am until 4:00 pm., on electron diffraction in TEM.

Featured speakers will be Dr. Stavros Nicolopoulos, President of NanoMEGAS SPRL, Brussels Belgium;
Dr. Amith Darbal, Applications Scientist AppFive, LLC Tempe, Arizona; and Prof. Jorg Wiezorek, Univ
of Pittsburgh Dept of Mech Engr and Matl Sci Swanson School of Engr, Pittsburgh, PA. Talks will
cover recent results using precession electron diffraction in TEM, and be followed by a
demonstration of various applications.

In order to register, please RSVP by email at sergey-at-seas.ucla.edu. Respond no later than Friday
October 25th.

Lunch will be provided. The event is open to all.

Please pass this notice along to those you feel would be interested.

We hope to see you at the workshop.

http://www.ms.ucla.edu/events/events-archive/event-flyers/2013/electron-diffraction-workshop.pdf/

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Email: mohamed.jaffer-at-uct.ac.za
Name: Mohamed Jaffer

Organization: University of Cape Town

Title-Subject: [Filtered] Vacancy: Principal Technical Officer: Electron Microscope Unit, UCT

Message: Dear All

The University of Cape Town(UCT) Electron Microscope Unit (EMU) has a job opening for a Principal
Technical Officer.

The EMU provides a service to all faculties at UCT, as well as external users. The Unit offers
services on a number of electron beam platforms viz., a FEI Tecnai F20, a FEI Tecnai 20 with Tridiem
energy filter, a FEI Nova Nano SEM, a Zeiss 912, a Leica S440 and a field emission QEMSCAN. In
addition, the EMU has a suite of associated equipment for sample preparation.

The overall purpose of the EMU Principal Technical Officer is to maintain, service and repair the
equipment of a world-class EMU at UCT.

The full advertisement can be viewed at:

http://www.uct.ac.za/about/intro/vacancies/external/

Thanks
Regards
Mohamed Jaffer

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Research Scientist(s) position
Please see the link below for the job information and to apply electronically online:

https://jobs.illinois.edu/academic-job-board/job-details?jobID=36125&job=research-scientist-materials-research-laboratory-a1300472

*Note for those unfamiliar with University jobs here, a common question: Is the job for only one year with the "12 month appointment"?

All academic positions at the University are appointed on a 9- or 12-month service basis, eligible for annual renewal based upon mutual agreement and the annual performance review process. This Academic Professional position is a regularly recurring staff scientist role not subject to grant funding. See http://ap.illinois.edu for more information.

{ { { { { { { { { { { { { { { { { {} } } } } } } } } } } }
Lou Ann Miller
Frederick Seitz Materials Research Laboratory
Lab Room 125 Office 374 MRL
104 South Goodwin Ave
Urbana, IL 61801
Campus mail code: MC-230

217-244-1567
lamiller-at-illinois.edu
http://mrl.illinois.edu

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Warren, The frequency component of your issue makes me wonder if you simply have an aging power supply. 120HZ is the frequency one sees (in the Americas) at the output of the power supply rectification and it's not uncommon for the typical wrapped foil-electrolyte capacitors to age and dry out with time, losing their capacitance. A bulged capacitor would be a sure give-away of this problem. The ripple voltage at the main caps could be checked with an oscilloscope; the lowest voltage should still be several volts above the regulated output. Even without schematics, most older supplies are linear designs and a capable technician can identify the circuitry. Without a scope, an inexpensive attempt at repair would be to simply replace all the electrolytic capacitors in the supply. This can typically be done for less than $20 in parts. Hope this helps. Rick

Richard A. Ross
Allison Transmission Inc.
Indianapolis, IN USA

-----Original Message-----
X-from: wesaia-at-iastate.edu [mailto:wesaia-at-iastate.edu]
Sent: Wednesday, October 23, 2013 6:00 PM
To: Richard A. Ross

We are running an older Oxford Tetra(tm) backscattered electron system on our older Hitachi S-2460N SEM. The SEM has been a trooper through the years, but the Tetra BSE system has recently developed some nasty noise in the signal.

It is scan rate dependent and seems to track back to 60 Hz. If we enable the Sync-to-AC feature we get nice vertical lines on our screen. When we turn it off, we get a splattering of bright dots across the captured image. It appears to be 8.3 ms between lines which corresponds to 120 Hz or every zero crossing of the AC line.

Does anyone have some spare Tetra pieces around that they are willing to part with? Our system has the external box (PN 1108-091) that is controlled by a serial connection from the EDS computer. I suspect the problem is in that box, but it might be in the detector assembly/pre-amp.

I have posted pictures of our unit and some problem images on our lab server at ftp://www.marl.iastate.edu/equipment/tetra.

Looking again to this helpful community, Warren Straszheim Assoc. Scientist Iowa State University
515-294-8187

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Listers,

I am looking for ways to stain composites of polyvinyl alcohol containing
cellulose nano fibrils for TEM or back-scattering SEM. The samples are
films that need to be embedded and sectioned for TEM. Information required
would be the orientation and distribution of the nano fibrils in the
polyvinyl alcohol composite. Any preparation suggestions would be very
welcome.

Debby

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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7, 30 -- From: "Sherman, Debra" {dsherman-at-purdue.edu}
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Please share the following announcement with those at your institution
who might be interested.

The NRI-MCDB Facility at UC, Santa Barbara is hosting an Advanced
Microscopy and Digital Imaging Workshop Jan 14-17, 2014. Participants
who will benefit most from the workshop are newer to light microscopy
and are interested in learning more about transmitted light and
fluorescence microscopy including confocal.

Advanced Microscopy and Digital Imaging Workshop Jan 14-17, 2014.
https://www.lifesci.ucsb.edu/mcdb/events/imaging_workshop/

Best Regards
Mary

--
Mary Raven
Microscopy Facility Director
Neuroscience Research Institute &
Molecular, Cellular & Developmental Biology
http://microscopy.nri.ucsb.edu
Phone: (805) 893 8702

Mail to:
NRI, Biology 2 Building, room 5173
The University of California
Santa Barbara, CA 93106-5060

2014 Advanced Microscopy and Digital Imaging Workshop
https://www.lifesci.ucsb.edu/mcdb/events/imaging_workshop/
Jan 14- Jan 17, 2014

==============================Original Headers==============================
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9, 20 -- Subject: LM: Advanced Microscopy and Digital Imaging Workshop Jan 14-17,
9, 20 -- 2014 in Santa Barbara, CA
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Hello!

Our 2 Day biological tutorial fall conference , "How Do We Do That?", November 13-14th, is approaching the end of the early registration deadline, October 31st.

Cost includes food, vendors, lectures, tours and demonstrations: $45 early registration, $55 late registration.

This program covers topics from Cryo, Immuno techniques, forensics, Fluorescence etc. Vendors will also be present, and include, JEOL, Hitachi, Gatan, RMC & Boeckeler Instruments, Tousimis, and ibidi, LLC

( New vendors, it is not too late for you to register please do so by Friday)

Our Program: http://mrl.illinois.edu/sites/default/files/pdfs/BioStructures2013Program.pdf

Registration, and more conference information: http://mrl.illinois.edu/events/conferences-workshops/second-annual-biological-imaging-workshop

Hope you can join us! This is perfect for staff and students, both biological, and those in material sciences that mix the two disciplines.

Lou Ann

{ { { { { { { { { { { { { { { { {} } } } } } } } } } } } } } } }
Lou Ann Miller
Biological Electron Microscopy
Frederick Seitz Materials Research Laboratory
Lab Room 125 Office 374 MRL
104 South Goodwin Ave
Urbana, IL 61801
Campus mail code: MC-230

217-244-1567
lamiller-at-illinois.edu
http://mrl.illinois.edu

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Announcing: a *Northern California Society for Microscopy Meeting on
Tuesday, November 12th*
1:00 PM -- ~4:30 PM at
Berkeley Arts Festival
2133 University Ave
Berkeley CA

just a few blocks from UC Berkeley campus and BART

Presentations include:
Immuno Electron Microscopy in Neuroscience Studies
Prof Ida Llewellyn-Smith
Cardiovascular Medicine and Physiology School of Medicine
Flinders University
Bedford Park SA, AUSTRALIA

A Micro/Macro Texture Analysis of Anatase Films with Preferred
Orientation of {001} Facets
Matt Sanchez
Dept of Chemistry and Biochemistry, San Francisco State University
Advisor- Andrew Ichimura

3D Correlative Microscopy Employing Light, X-Ray and Electron Insturments
Kevin Fahey et al, Carl Zeiss Microscopy

==============================Original Headers==============================
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X-from: zhang_zaoli-at-yahoo.com ()

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Email: zhang_zaoli-at-yahoo.com
Name: zaoli Zhang

Organization: Erich Schmid institute

Title-Subject: [Filtered] Does the installation of a big telescreen in the microscope room affect
the stability of spectrometer and Cs corrector ?

Message: Dear Colleagues

Since 2009, one Cs-corrected microscope with Tridiem system was installed in our institute. From
time to time,we have to give Demo to some politiciens and guests. As the computer screen is a bit
small, we tend to install one big TV screen in the microscope room. However, we worry a bit about
the side-effects which may come with the installation of telescreen, such as cables and other
electronic devices.... particularly influence the stability of spectrometer or stability of Cs
corrector.

Does anyone know about this ? please give us some suggestions.

Best regards,

Zaoli Zhang

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EM senior technician in Yale Medical School

We are seeking a motivated and experienced EM technician to join our team
in the EM Core Facility. The primary duties are to process, embed and
plastic section biological samples. The ability to view specimen with
users in electron microscope is desirable but not necessary. Prior
experience with cryosectioning and immunoEM are plus. Please contact me
directly if you are interested or have questions.

The starting date and nature of the job (part-time vs full-time) are
negotiable.

Xinran Nick Liu, M.D. & Ph.D.
Director of Electron Microscopy Core Facility

Yale University School of Medicine333 Cedar Street, SHM I-E16A
New Haven, CT 06520
Office: 203.785.4050
http://medicine.yale.edu/ccmi/em

==============================Original Headers==============================
13, 35 -- From xinran.liu-at-yale.edu Tue Oct 29 12:52:30 2013
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13, 35 -- Oct 2013 13:52:29 -0400
13, 35 -- From: "Liu, Xinran" {xinran.liu-at-yale.edu}
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13, 35 -- Subject: EM technician postion - Yale Medical School
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Dear all,

Today I noticed a problem in the Auriga Zeiss SEM-FIB microscope that I
daily work.
The nozzle temperature of the GIS system remains at 44ºC, and doesnt go
higher. Normally takes 10 min to reach ~80ºC. The bakeoff took 3h to
increase 6ºC.

Does someone have ever experienced any similar problem?

I had an electricity problem during the night and the microscope went
off for all night. I do not know if this situation is even related to
the FIB problem.

Thank you for all help.

Regards.

Daniela Gomes.

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Name: David Lowry

Organization: Arizona State University

Title-Subject: [Filtered] Thiery silver proteinate stain

Message: we're having a problem using a variation of the Thiery technique in an attempt to stain
glycogen on thin sections for TEM. The staining is actually working OK but we are getting very bad
silver protein precipitation on the section surface in the form of web-like masses as well as
aggregates of discrete colloid-spheres. We are using the silver proteinate at 1% aqueous and have
tried dissolving 24 hr in advance, centrifuging at max speed in tabletop centrifuge, and 0.2 um
polymer filtration through syringe prior to incubation (all kept dark). Extensive washing with
di-water following silver incubation does not help.

Does silver proteinate have a definite shelf-life beyond which it produces these sorts of problems?
Our dry silver reagent is several years old but has been kept wrapped in foil at room temp and looks
good, no moisture or clumping in the vial. Any advice would be much appreciated.

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Dear David -ALL,

it is "refreshing" for me as a kind of {dinosaur in EM} that such old and ancient/ancillary methods are used nowadays too.
That there might sometimes problems: it will not be the last time I guess..

for me first of all some questions:

1) "di-water" means "deionized" or "distilled" water??

2) if "di-water" is / means { distilled water } then:
a) outside source (e.g. pharmacy) or b) selfmade (= self controlled production)

if a) do you know the quality of the water (i.e. perhaps higher amount of metal-ions like Cu due to distillation source).

This concern might be applicable to the making of Ag-Proteinate solution as well as the following washing steps.

3) Do you know about the cleanliness of your prep-equipment, esp. the 0.2 �m filters used.?
4) Do you use regular Cu-grids or Gold-grids ?

Unfortunately no knowledge about shelflife of silver proteinate, but in mind that dissolving Ag-proteinate has been done also by ultrasonication.

Sorry if this doesn't help you anyway,
very best regards and good luck,

Wolfgang

Wolfgang MUSS PhD
EM-Lab, Pathology
SALK-LKH (Gen. Hosp. and PMU=priv. Paracelsus Med.Univ. SALZBURG)
SALZBURG, AUSTRIA

PS: cf.
Coulary - Aigle - Schaeffer: J Electron Microsc*) (Tokyo)-50(2)/2001
* The Title of this particular Journal has changed some time ago to "MICROSCOPY"

Journal of Electron Microscopy, Volume 50, Issue 2, pp. 133-137: Abstract.
Evidence for glycogen structures associated with plasma membrane invaginations as visualized by freeze-substitution and the Thiery reaction in Saccharomyces cerevisiae
B Coulary, M Aigle* and J Schaeffer
IBGC UMR 5095, 1 Rue Camille Saint-Sa�ns, F-33077 Bordeaux Cedex, France,
*Corresponding author, E-mail: Michel.Aigle-at-ibgc.u-bordeaux2.fr
Abstract:
We have used a combination of freeze-substitution electron microscopy and specific reaction for polysaccharides to re-evaluate glycogen structures in Saccharomyces cerevisiae. We also used mutant cells devoid of glycogen to confirm the glycogenic nature of the structures described. Previously described cytoplasmic aggregates were confirmed as a glycogen granules. Moreover, an original structure was discovered. This is a ring of glycogen surrounding a finger- or pleat-like plasma membrane invagination. This structure could be physiologically significant in terms of channelling glucose to or from glycogen reserves.

Keywords: glycogen, yeast, Thiery reaction, plasma membrane, invaginations glycogenin, Saccharomyces cerevisiae, freeze-substitution

ex methods: { {....Gold grids containing thin sections were treated with periodic acid 1% for 20 mins at room temperature and rapidly rinsed in water twice and for 3 x(times) 10 mins. Samples were then placed in a solution of thiocarbohydrazide 0.2% in acetic acid 20% for 30 mins in a dark room. Grids were rinsed in acetic acid 10% (twice rapidly, and 2 x[times] 7 mins) followed by 3 x(times) 7 mins in water. Polysaccharides were revealed by incubating the samples in silver proteinate 1% in water for 30 mins in the dark. Grids were then rapidly rinsed in water and observed in the EM.} }

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} Gesendet: Mittwoch, 30. Oktober 2013 13:41
} An: Mu� Wolfgang
} Betreff: [Microscopy] Thiery silver proteinate stain
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} Name: David Lowry
} Organization: Arizona State University
} Title-Subject: Thiery silver proteinate
} stain
} Message:
}
} we're having a problem using a variation of the Thiery technique in an
} attempt to stain glycogen on thin sections for TEM. The staining is
} actually working OK but we are getting very bad silver protein
} precipitation on the section surface in the form of web-like masses as
} well as aggregates of discrete colloid-spheres.
}
} We are using the silver proteinate at 1% aqueous and have tried
} dissolving 24 hr in advance, centrifuging at max speed in tabletop
} centrifuge, and 0.2 �m polymer filtration through syringe prior to
} incubation (all kept dark).
} Extensive washing with di-water following silver incubation does not
} help.
}
} Does silver proteinate have a definite shelf-life beyond which it
} produces these sorts of problems?
}
} Our dry silver reagent is several years old but has been kept wrapped
} in foil at room temp and looks good, no moisture or clumping in the
} vial. Any advice would be much appreciated.
}
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Dave,
I have no experience with this technique so cannot help you with it.
However, you might consider a alternative technique next time this comes
up. This is the PATO technique utilizing ruthenium red to stain
polysaccharides. It has worked very well for us in the past identifying
glycogen deposits in cyanobacteria (see references below). You can
download the technique from my website.

Debby

J.S. Hanker, etal. (1964). Osmophilic Reagents: New Cytochemical Principle
for Light and Electron Microscopy. Science
146:1039-1043.

Sherman, Debra M. and Louis A. Sherman. (1983). Effect of iron deficiency
and iron restoration on
ultrastructure of Anacystis nidulans. J. Bacteriol. 156:393-401.

Debra Sherman, Founder & CSO
DS imaging LLC
Purdue Technology Center
1281 Win Hentschel Blvd
West Lafayette, IN 47906
E-mail: debby.sherman-at-dsimagingllc.com
www.dsimagingllc.com
Mobile: 765-418-8540

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Email: rfoley-at-uab.edu
Name: Robin Foley

Organization: UAB

Title-Subject: [Filtered] ESEM - drying biological samples!

Message: All-

Background: I'm a metallurgist with lots of high vac. SEM experience but very limited ESEM.

Question: I've got biological types bringing in samples like rat hearts for ESEM. They bring the
samples soaking in alcohol or water. The samples are hydrated and plump.

We do ESEM on the samples (temperature and humidity control). I start the water vapor pressure high
enough that water condenses on the samples in the ESEM and then turn it down just until the liquid
on the surface goes away.

When the samples come out - they look like damp leather.

Does it sound like I am doing this right? Should the samples come out of the ESEM looking as plump
and hydrated as when they went in?

Thanks,
Robin

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Email: Dan.Buntman-at-wdc.com
Name: Dan Buntman

Organization: Western Digital

Title-Subject: [Filtered] Sr. Principal Engineer position-TEM and Media specialist

Message: We currently have the following opening at our San Jose R & D facility:

Sr. Principal Engineer TEM Specialist for Magnetic Media-06745
Description
Successful candidate will utilize TEM expertise to perform advanced evaluation of R&D disk drive
media products and develop new capabilities to support future R&D needs.
Will interact with magnetics research engineers to understand development goals and needs, and will
supply necessary structural information to meet technical, quality, and turnaround requirements.
Will be responsible for equipment uptime, including interfacing with equipment vendor, calibration
and maintenance of the TEM.
Will lead in developing/refining/documenting advanced TEM/STEM/EDX/EELS techniques and training
other members of the Materials Analysis group.
Must be able to work in a fast-paced, multi-tasking environment where priorities may change
unexpectedly, and sustain positive interactions with others.
Qualifications
Education: PhD in materials science or closely related discipline. Excellent verbal and written
communication skills required.

Experience: 6 or more years experience in developing/refining advanced TEM techniques. Hands-on
and expert in all operational aspects of TEM (BF/DF/STEM/EDX/EELS/nano-diffraction). Background in
materials science and thorough understanding of advanced scientific principles and technical
advances of the magnetic recording industry and media processes. FIB and manual polish expertise a
plus.

To apply online go to:
http://www.wdc.com/en/company/employment/northamerica.aspx

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Email: lea.sabina-at-yahoo.de
Name: leas

Organization: university bern, switzerland

Title-Subject: [Filtered] LM, need help on staining cells/CD31/rat

Message: Hello

I'm currently working with the anti-rat cd31, clone TLD-3A12 antibody for immunohistochemical
staining of rat formalin fixed paraffin tissue. Unfortunately so far there is no positive outcome.
Has anyone experience with this antibody or recommendations for a different antibody?

Thanks a lot for your help

Lea

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Can anyone point me in the direction of a clear ray diagram of how a TEM Cs corrector works?

I�ve had a good look around on the web and can�t find what I want:

1. Some of the ray diagrams are just plain wrong, in my opinion ...

2. Some are complex but don�t actually show what they claim to show,

3. Many simply avoid the issue by having a rectangular box with no detail �.

I appreciate that the actual electron paths are complex multiple helices but personally, I find ray diagrams quite informative in understanding the basics of what is going on inside a TEM. A simple ray diagram can, for example, clearly illustrate how spherical abberation arises so it should be possible to illustrate the correction of Cs.

Now, it could be that I�m trying to over complicate the question and it really is very simple - a Cs corrector, in ray diagram terms is acting very much like a concave glass lens does for light?

On the other hand, I have seen diagrams showing that Cs correctors are not rotationally symetric about the optic axis - which is why they come in pairs so it isn�t quite as straight forward as a concave glass lens.

Thank you,

Best Regards,

Larry Stoter
EO Sales & Business Development General Manager
��
JEOL (UK) Ltd
Silver Court, Watchmead, Welwyn Garden City, Herts, AL7 1LT, United Kingdom
Tel: +44-(0)1707-377117 Fax: +44-(0)1707-373254 Mobile: +44-(0)7967-388617
E-mail: larry.stoter-at-jeoluk.com Web: {http://www.jeoluk.com/}
��

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Email: info-at-nesmicroscopy.org
Name: New England Society for Microscopy

Organization: New England Society for Microscopy

Title-Subject: [Filtered] Save the Date: NESM's annual Fall Symposium & Business Meeting at Brandeis
University!

Message: Dear Fellow Microscopists,

This is a friendly reminder to save the date for the New England Society for Microscopys annual
Fall Symposium & Business Meeting on Thursday, December 5th at Brandeis University. The Symposium
will consist of four technical talks, a coffee break, a buffet dinner, and our annual business
meeting and board elections. Stay tuned more information to come in the following weeks! Keep up
with NESM by visiting our website (www.nesmicroscopy.org) and by following us on Facebook
(http://www.facebook.com/NESMicroscopy) and on twitter (http://www.twitter.com/#!/NESMicroscopy).

Cheers,
Rylie Walsh
NESM Corresponding Secretary 2014

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Dear All,
we will be running our Requimte AFM training workshop again this Easter 2014. It will take place in Porto, Portugal. The course will be four days between�14th to 17th April, and include background lectures, lots of practical information, hands-on time, and a data processing class, all on atomic force microscopy. The target audience is basically all beginners in atomic force microscopy, from surface science to biology. In the past, the course has gotten great feedback from the students. For more details, just go to�http://bit.ly/T0uO22.
Regards,
Pete.

____________________________________________________________________________
Atomic Force Microscopy
Dr Peter Eaton and Dr Paul West
Available through all good bookshops, or direct from Oxford University Press at:
http://ukcatalogue.oup.com/product/9780199570454.do http://afmhelp.com

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Hi, everyone!

We still keep our JEOL TEM 4000 running and it works well. At this moment we are still using films and we are having problems on film camera.

The camera on 4000 recently jams the film cassettes a lot. Therefore I am thinking to get help from this listserver.

Since there are a lot of JEOL TEM machines around in USA and the world, and all the film cameras are the same basically, also as I know a lot of people install CCD cameras on these JEOL TEM and start to give up using films.� �I guess there would be some cameras which are not in use. Therefore I am asking if you have JEOL camera(s) not in use and are willing to give to us or sell to us, it would be great. We will cover the cost.

Also we are looking for cassettes for TEM 4000. Each cassette for TEM 4000 = does not have a back since higher accelerating voltage (400KV). And it is different from 200KV or 100KV�� machines.� If you have some cassettes which are for high HT, please let me know too. We do not need any cassettes with backs. We have some ourselves.

I would say thanks in advance.

Have a nice day!

Zhenquan Liu

Engineer Principal
Arizona State University

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Ondrej,

Thank you - strangely, attachment with your mail came through OK but the listserver keeps rejecting my reply for having an attachment - which it doesn�t �.

Not being in a university makes it difficult to do proper literature searches without spending a small fortune!

That�ll give me something to read this November weekend while the rain comes down ...

Larry

On 01 Nov 2013, at 00:27, Ondrej Krivanek {krivanek-at-nion.com} wrote:

} Hi Larry,
}
} We wrote a chapter on aberration correction some 5 years ago. It being an early text, it makes sure not to skip anything that more recent texts may take for granted. In other words, I believe that it explains the principles of aberration correction pretty well. It focuses primarily on STEM, but reversing the flow of the electrons in the diagrams (reciprocity does work!) so that they apply to CTEM applications should not be too much of a hardship.
}
} I've attached the chapter to this email. The list server will probably not let it through, so here's the full reference:
}
} Krivanek O L, Dellby N, and Murfitt M F (2009) Aberration correction
} in electron microscopy. In: Orloff J (ed.), Handbook of
} Charged-Particle Optics, pp 601�640 (CRC Press, Boca Raton).
}
} Cheers.
}
} Ondrej
}
}
} On Thu, Oct 31, 2013 at 1:47 AM, {larry.stoter-at-gmail.com} wrote:
}
}
}
} ----------------------------------------------------------------------------
} The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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}
} Can anyone point me in the direction of a clear ray diagram of how a TEM Cs corrector works?
}
} I�ve had a good look around on the web and can�t find what I want:
}
} 1. Some of the ray diagrams are just plain wrong, in my opinion ...
}
} 2. Some are complex but don�t actually show what they claim to show,
}
} 3. Many simply avoid the issue by having a rectangular box with no detail.
}
} I appreciate that the actual electron paths are complex multiple helices but personally, I find ray diagrams quite informative in understanding the basics of what is going on inside a TEM. A simple ray diagram can, for example, clearly illustrate how spherical abberation arises so it should be possible to illustrate the correction of Cs.
}
} Now, it could be that I�m trying to over complicate the question and it really is very simple - a Cs corrector, in ray diagram terms is acting very much like a concave glass lens does for light?
}
} On the other hand, I have seen diagrams showing that Cs correctors are not rotationally symetric about the optic axis - which is why they come in pairs so it isn�t quite as straight forward as a concave glass lens.
}
} Thank you,
}
} Best Regards,
}
} Larry Stoter

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Organization: Oxford University, Department of Materials

Title-Subject: [Filtered] Two Post-doc positions available

Message: Applications are invited for two postdoctoral positions one each in atom probe tomography
and transmission electron microscopy to examine radiation effects in perovskites. The researchers
appointed to these posts will be expected to undertake and facilitate advanced nanostructural and
chemical characterisation studies of radiation damage formation and recovery mechanisms, and inert
gas incorporation in a range of perovskite materials for nuclear energy (fission and fusion)
applications. These posts are available for up to three and a half years duration from 1 March 2014
and are part of an EPSRC funded collaboration between the Universities of Oxford, Sheffield,
Huddersfield and Imperial College London, and including industrial partners from the Culham Centre
for Fusion Energy and Tokamak Solutions.

Applicants should have, by the start date, a doctorate in Physics, Materials Science, Engineering,
Chemistry or a closely-related subject area. Proven skills are required in some or all of electron
microscopy, atom probe tomography, and focussed ion beam machining, preferably as applied to
investigating the effects of radiation damage in materials. Also required are excellent written and
oral communication skills, willingness to travel to liaise with collaborators within the UK and
internationally when required, and the ability to assist with research management, including the
training of junior researchers.

Informal enquiries may be addressed to Dr Phil Edmondson (email: phil.edmondson-at-materials.ox.ac.uk).
Information on nuclear materials research being conducted at Oxford can be found either at
http://mffp.materials.ox.ac.uk or http://www-fim.materials.ox.ac.uk.

The closing date for applications is midday on Monday 6 January 2014 with interviews currently
planned for the week beginning 20 January 2014.

Applications for this post are to be made online (vacancy reference 110342). To apply for this role
and for further details, including the job description and selection criteria, please click on the
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Email: chris.brantner-at-nih.gov
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We are seeking an entry-level EM technician to join the NHLBI EM Core Facility staff on the
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preparative instrumentation. There is the opportunity to perform cryosectioning and immunogold
labeling, so these skills are also desirable. Please contact me directly if you are interested or
have questions about this position.

This is a contract position. This is a full-time position with a start date as soon as possible.

Chris Brantner, Ph.D.
Deputy Director of the NHLBI Electron Microscopy Core Facility
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National Institutes of Health
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Title-Subject: [Filtered] PhD scholarship in Nanoscale Imaging of the Aqueous Processes of
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The Department of Micro- and Nanotechnology invites applications for a 3-year PhD Scholarship in
Nanoscale Imaging of the Aqueous Processes of Precipitation, Dispersion, and Imbibition.

Heterogeneous processes in aqueous environments are fundamental for a wealth of industrial and
natural processes: Precipitation, imbibition, and dispersion of both liquid and solid phases in
water. These processes are very difficult to image as they are initiated on the nanoscale.

The aim of this project is to develop new nanoscale imaging methods with emphasis on microfluidic
chip-based methods and electron microscopy, for imaging of these processes in model systems and to
test their performance compared to other imaging methods and relevant samples from industrial
collaborators.

The two main application cases are: Precipitation and imbibition processes involving oil and water
in nano- and micrometer sized channels/pores relevant for enhanced oil recovery, and nucleation and
precipitation processes forming dispersions of particles and slurries with relevance for catalysis.

The project is jointly financed by DTUs Department of Micro- and Nanotechnology (DTU Nanotech,
www.nanotech.dtu.dk ), the Centre for Electron Nanoscopy (CEN, www.cen.dtu.dk ), and the Center for
Energy Resources Engineering (CERE, www.cere.dtu.dk ).

Responsibilities and tasks
The Ph.D. project will involve electron microscopy at DTU advanced microscopy facility CEN, the use
and development of cryogenic freezing equipment, cleanroom fabrication of microchip systems in our
1300m2 cleanroom facility, and performing experiments and detailed analysis of the chemical and
physical processes with the project partners.

A background in chemistry and/or physics and experience with the above methods will be beneficial.

Qualifications
Candidates should have a master's degree in engineering or a similar degree with an academic level
equivalent to the master's degree in engineering.

Further information
Further information may be obtained from Kristian M�lhave, Head of the Molecular Windows group at
DTU Nanotech on Kristian.molhave-at-nanotech.dtu.dk or tel.: +45 2512 6672.

For more information and submission of application please see:
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Title-Subject: [Filtered] Third Party Repair Service for Old Microtomes

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Hello Listserver,

The motor drive belt on our Ultracut E just failed after many long years of faithful service. I've managed to find a replacement belt, (it's actually just an o-ring) but am a bit stumped on how to remove the hand wheel to be able to slip the new belt on the drive pulley. I can get the plastic cover off, but it looks like I might need a special tool to get further into the assembly. Has anyone out there done this themselves, or is there a repair manual available?

Thanks,
Bradford Ross

Microscopy Technician

BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996

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Hello again everyone,

With some more concerted (and colleague consulted) fiddling, I managed to figure it out! Paul Webster hit the nail on the head saying that it has to do with the position of the hand wheel. The red dot on the cutting window adjustment part of the wheel needs to be lined up with the motor switch lever, and then you just turn the drive pulley until the wheel slides off. Then you just loop the belt around the drive shaft and shove it through the small notch in the microtome case and slip the belt onto the drive pulleys. I took pictures of the whole process in case anyone is interested.

Thanks,
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996

________________________________________
X-from: bradford.ross-at-botany.ubc.ca [bradford.ross-at-botany.ubc.ca]
Sent: Monday, November 04, 2013 3:34 PM
To: Ross, Bradford

Hello Listserver,

The motor drive belt on our Ultracut E just failed after many long years of faithful service. I've managed to find a replacement belt, (it's actually just an o-ring) but am a bit stumped on how to remove the hand wheel to be able to slip the new belt on the drive pulley. I can get the plastic cover off, but it looks like I might need a special tool to get further into the assembly. Has anyone out there done this themselves, or is there a repair manual available?

Thanks,
Bradford Ross

Microscopy Technician

BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996

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Email: alfredotolley-at-hotmail.com
Name: Alfredo Tolley

Organization: Instituto Balseiro - CNEA

Title-Subject: [Filtered] cannot remove HAADF detector

Message: We have recently installed a Tecnai F20 that has a HAADF detector and a wide angle CCD
camera that share the 35 mm port.

The HAADF detector was inserted to obtain Z contrast STEM images and now cannot be removed.

If we click to insert the CCD camera using TIA, the HAADF detector is properly removed first and
then the camera is inserted. On removing the camera, the HAADF detector is automatically re-inserted
and does not respond to software indications of removal.

I am writing in the hope than someone might have run into this kind of problem before and may know
the way to solve it.

Many thanks!

Alfredo Tolley

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Listers,
We have encountered a problem with LR White polymerization using UV light. We have an immuno protocol that requires -20C polymerization with UV after a PLT protocol.
The resin does not polymerize at -20C, nor 4C, nor room temp (well maybe a little at RT) using UV. It polymerizes fine at 50C in an oven.

Thoughts/suggestions?

Many thanks

Chris

Christopher J. Gilpin Ph.D.
Director, Life Science Microscopy Facility
Purdue University
Whistler Hall of Agriculture Research, Room S052
170 S. University St
West Lafayette, IN 47907
765-494-7750
gilpin-at-purdue.edu
http://www3.ag.purdue.edu/facilities/microscopy/Pages/default.aspx

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7, 28 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
7, 28 -- Subject: LR White not polymerising with UV
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Hi Chris,

The times when LRW polymerization has given me conniptions generally involve 1) exposure to oxygen during polymerization, 2) outdated resin, and 3) residual amounts of osmium in the sample (this latter happens when LRW is used for other reasons than immuno work, such as wanting some specific cutting properties).

1) When using flat-embedding molds, make sure that the blocks you value are in the middle of a row of "dummies", all filled to slightly overflowing, then covered with suitably sized cover slip (Thermonox or glass). The outside blanks tend to act as buffers for the inside specimen blocks.

2) I am suspicious of LRW much over a year old. I tend to use older bottles for infiltration up until the last couple changes, but use newer resin for the last exchanges.

3) I have had LRW turn to the consistency of cottage cheese when there were trace amounts of OsO4 left in the tissue. Had me puzzled big time until Dr. Tom Phillips said "Aha!!" and set me straight.

If I think of anything else, I will shout "Aha!!" and pass it along.

Good luck,
Randy

Randy Tindall
Senior EM Specialist
Electron Microscopy Core Facility
W122 Veterinary Medicine
University of Missouri
Columbia, MO 65211

Phone: 573-884-5383
Email: tindallr-at-missouri.edu
Day job: www.emc.missouri.edu
Gallery: https://www.facebook.com/RandyTindallGalleries/timeline
Blog: http://nadiasyard.com

-----Original Message-----
X-from: gilpin-at-purdue.edu [mailto:gilpin-at-purdue.edu]
Sent: Tuesday, November 05, 2013 9:07 AM
To: Tindall, Randall D.

Listers,
We have encountered a problem with LR White polymerization using UV light. We have an immuno protocol that requires -20C polymerization with UV after a PLT protocol.
The resin does not polymerize at -20C, nor 4C, nor room temp (well maybe a little at RT) using UV. It polymerizes fine at 50C in an oven.

Thoughts/suggestions?

Many thanks

Chris

Christopher J. Gilpin Ph.D.
Director, Life Science Microscopy Facility Purdue University Whistler Hall of Agriculture Research, Room S052
170 S. University St
West Lafayette, IN 47907
765-494-7750
gilpin-at-purdue.edu
http://www3.ag.purdue.edu/facilities/microscopy/Pages/default.aspx

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7, 28 -- To: "Microscopy-at-microscopy.com" {Microscopy-at-microscopy.com}
7, 28 -- Subject: LR White not polymerising with UV
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Title-Subject: [Filtered] CSMMS Fall 2013 Meeting

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Greetings Fellow Microscopists,

The Midwest Microscopy and Microanalysis Society will hold its Fall meeting on Friday, November 22nd, at Baxter Healthcare Corporate Headquarters in Deerfield, IL. This meeting is a showcase for our Sustaining Members and Exhibitors, and they are providing an excellent lineup of presentations on the latest advances in techniques and instrumentation. MSA President and Tour Speaker Ernie Hall of GE Global Research will also be joining us. Please visit our website and follow the link to Meetings for the full program and other details:

http://www.midwestmicroscopy.org/

We look forward to seeing you there!

Elaine Schumacher
M3S Program Coordinator

*********************************************************************�
Elaine F. Schumacher
Senior Research Scientist
McCrone Associates, Inc.
850 Pasquinelli Drive
Westmont, IL� 60559-5539 USA
630-887-7100 (tel)
630-887-7417 (fax)
E-mail:�� eschumacher-at-mccrone.com
Web Site:� www.mccrone.com

*********************************************************************
This message and any attachments are solely for the��
intended recipient. If you are not the intended recipient,�
disclosure, copying, use or� distribution of the information
included in this message is prohibited.
*********************************************************************�

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Hello one more time Listserver,

I made a pdf of the process as I went through it. If anyone wants to download it, I threw it up on my Google Drive for anyone with this link to access it: (I may not leave it there for eternity, so get it while it's hot!)

https://drive.google.com/file/d/0B3suda2jASIMbFBTdnhkN3dYRjQ/edit?usp=sharing

Thanks to Wolfgang Muss for sending me a copy of the Ultracut service manual. The way the manufacturer tells you to do this is slightly different from my method, but they don't document the process visually very well, so I think my document may still be of some use to people.

Cheers,
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996

________________________________________
X-from: Ross, Bradford
Sent: Monday, November 04, 2013 5:24 PM
To: Microscopy-at-microscopy.com

Hello Listserver,

The motor drive belt on our Ultracut E just failed after many long years of faithful service. I've managed to find a replacement belt, (it's actually just an o-ring) but am a bit stumped on how to remove the hand wheel to be able to slip the new belt on the drive pulley. I can get the plastic cover off, but it looks like I might need a special tool to get further into the assembly. Has anyone out there done this themselves, or is there a repair manual available?

Thanks,
Bradford Ross

Microscopy Technician

BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996

==============================Original Headers==============================
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13, 34 -- Subject: Reichert Ultracut E drive belt replacement
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Dear Friends

While viewing an epithelium in SEM I came across numerous spherical objects
of 10-20 microns size with their surfaces forming ruffles (please click this
link http://www.eikonika.net/v2/download.php). These ruffles remind me of
the appearance of mucous I have seen before.
Can anybody comment on the origin of these objects? Could they be epithelial
cells undergoing mucinous degeneration?

Thanks in advance to the wise List

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

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Those ruffles look too sharp-edged and close together, not like what
I've seen on cells. More like some minerals.
What was your fixative and buffer? My first thought is that the
spheroids are precipitates. If you have an EDS with your SEM, an
analysis of a couple of spheroids would be a good starting point.

Phil

} Dear Friends
}
} While viewing an epithelium in SEM I came across numerous spherical objects
} of 10-20 microns size with their surfaces forming ruffles (please click this
} link http://www.eikonika.net/v2/download.php). These ruffles remind me of
} the appearance of mucous I have seen before.
} Can anybody comment on the origin of these objects? Could they be epithelial
} cells undergoing mucinous degeneration?
}
} Thanks in advance to the wise List
}
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} Tel/fax +30 210 8957677
} mobile +30 6945 107477
} www.eikonika.net www.aim.cat
} *************************************
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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Hello Yorgos,
I think these structures might be complexes of phosphate with some
divalent cations (~ Ca2+ ???). Was the sample in phosphate buffer?

A long time ago I have been working on particles appearing in culture
media containing heavy metals. Here is the link to the SEM image how
they looked:
http://www2.biomed.cas.cz/~benada/image.html
The image shows Cd2+ complexes with phosphate (I really do
not know their stoichiometry).
We also were able to prepare them artificially by titration of 0.1M
phosphate buffer with cadmium sulfate.

My best regards from Prague

Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

On Wed, 6 Nov 2013 06:02:45 -0600, eikonika-at-otenet.gr wrote :
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} (please click this link http://www.eikonika.net/v2/download.php).
} These ruffles remind me of the appearance of mucous I have seen
} before. Can anybody comment on the origin of these objects? Could
} they be epithelial cells undergoing mucinous degeneration?
}
} Thanks in advance to the wise List
}
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} Dr Yorgos Nikas
} Athens Innovative Microscopy
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7, 39 -- From benada-at-biomed.cas.cz Wed Nov 6 07:24:03 2013
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Hi Yorgos,
for me it looks like beads from some Carbonate, like Calciumcarbonate.
Some years ago I took some images from precipitates in a watercooker here at Wuerzburg where we have very hard water and the
pattern of the crystals look the same.
Do you have EDS? If not, send the specimen and I have a look.

Best wishes,
Stefan

-----------------------------------------------------
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Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
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++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
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www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
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Am 06.11.13 13:04, schrieb eikonika-at-otenet.gr:
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} While viewing an epithelium in SEM I came across numerous spherical objects
} of 10-20 microns size with their surfaces forming ruffles (please click this
} link http://www.eikonika.net/v2/download.php). These ruffles remind me of
} the appearance of mucous I have seen before.
} Can anybody comment on the origin of these objects? Could they be epithelial
} cells undergoing mucinous degeneration?
}
} Thanks in advance to the wise List
}
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} Tel/fax +30 210 8957677
} mobile +30 6945 107477
} www.eikonika.net www.aim.cat
} *************************************
}
}
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7, 22 -- From stefan.diller-at-t-online.de Wed Nov 6 07:29:31 2013
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They are red blood cells.
Karen Bentley

Karen Bentley, M.S.
Director
Electron Microscope Research Core
Pathology & Laboratory Medicine
585-275-1954

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Wednesday, November 06, 2013 7:05 AM
To: Bentley, Karen

Hi All

I am only a biologist when others have no idea, I was born as a
metallurgist, but these must be red blood cells?

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: 06 November 2013 12:01
To: protrain-at-emcourses.com

I have seen similar objects when working with acid-etched bone. EDS gave me composition similar to calcium phosphate/hydroxyapatite. EDS/BSE should give you an idea what you have.
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Wednesday, November 06, 2013 6:01 AM
To: Dusevich, Vladimir

Dear Friends

While viewing an epithelium in SEM I came across numerous spherical objects of 10-20 microns size with their surfaces forming ruffles (please click this link http://www.eikonika.net/v2/download.php). These ruffles remind me of the appearance of mucous I have seen before.
Can anybody comment on the origin of these objects? Could they be epithelial cells undergoing mucinous degeneration?

Thanks in advance to the wise List

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

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This is the hydroxyapatite crystals I once saw in SEM, looks similar to
yours, so I suspect it's some kind of mineral crystals.

https://www.dropbox.com/s/4poy865y18kcl27/Hydroxylapatite.jpg

Xuanhao

-----Original Message-----
X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Wednesday, November 06, 2013 7:13 AM
To: sunxh163-at-gmail.com

Dear Friends

While viewing an epithelium in SEM I came across numerous spherical objects
of 10-20 microns size with their surfaces forming ruffles (please click this
link http://www.eikonika.net/v2/download.php). These ruffles remind me of
the appearance of mucous I have seen before.
Can anybody comment on the origin of these objects? Could they be epithelial
cells undergoing mucinous degeneration?

Thanks in advance to the wise List

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

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Based on previous responses I found this image in a manufacturers website, of hydroxyapatite. http://www.jeolusa.com/tabid/323/AlbumID/570-22/Default.aspx
Looks nearly identical to your image.

Dave

____________________________
David J. Wilbur, Ph.D.
Instrumentation Specialist
Department of Chemistry
Tufts University
62 Talbot Ave.
Medford, MA 02155
voice:�� 617-627-2163
Fax:�� 617-627-3443
email:�� david.wilbur-at-tufts.edu

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X-from: eikonika-at-otenet.gr [mailto:eikonika-at-otenet.gr]
Sent: Wednesday, November 06, 2013 7:03 AM
To: Wilbur, David J.

Dear Friends

While viewing an epithelium in SEM I came across numerous spherical objects of 10-20 microns size with their surfaces forming ruffles (please click this link http://www.eikonika.net/v2/download.php). These ruffles remind me of the appearance of mucous I have seen before.
Can anybody comment on the origin of these objects? Could they be epithelial cells undergoing mucinous degeneration?

Thanks in advance to the wise List

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

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16, 31 -- From David.Wilbur-at-tufts.edu Wed Nov 6 09:19:56 2013
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It's a really tough task - to observe soft tissue with ESEM.
You can try rewetting specimens every few minutes (change pressure so that you will see droplets of water on the specimen).
You can try fixation prior to observation.
Even better - perform all needed steps for conventional bio specimen preparation (fixation, dehydration, coating) and work in high vac mode.
Best of all - show pictures to your "biological types", and if they are happy, be happy.
Vladimir

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

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To: Dusevich, Vladimir

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Email: rfoley-at-uab.edu
Name: Robin Foley

Organization: UAB

Title-Subject: [Filtered] ESEM - drying biological samples!

Message: All-

Background: I'm a metallurgist with lots of high vac. SEM experience but very limited ESEM.

Question: I've got biological types bringing in samples like rat hearts for ESEM. They bring the samples soaking in alcohol or water. The samples are hydrated and plump.

We do ESEM on the samples (temperature and humidity control). I start the water vapor pressure high enough that water condenses on the samples in the ESEM and then turn it down just until the liquid on the surface goes away.

When the samples come out - they look like damp leather.

Does it sound like I am doing this right? Should the samples come out of the ESEM looking as plump and hydrated as when they went in?

Thanks,
Robin

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Hi All

Well I was wrong wasn't I, sorry I apologise as I did not scroll to the
right!

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
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-----Original Message-----
X-from: Philip Oshel [mailto:oshel1pe-at-cmich.edu]
Sent: 06 November 2013 15:14
To: protrain-at-emcourses.com

Hi everybody,

I'm learning to use CASTEP (or trying to) with EELS on the ionic materials
I work with. But I'm having a hard time going from the examples in
tutorials, which are all nice monatomic (semi)conductors like diamond and
silicon, to ionic insulators, where using an on-the-fly pseudopotential
seems to utterly fail every time.

Is there anybody out there who uses CASTEP by itself, not inside Materials
Studio? Do you have input files (.cell, .param) for anything ionic, that I
could look at and see where I'm going wrong/where I need to go at all?

NaCl would be FANTASTIC (it's the thing I'm using for practice before I
start on the real experiments) but anything would help. Please somebody
help, I have google-scholared the internet to death.

Jo Sharp

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The disc/biconcave disc-shaped structures are almost certainly red blood cells (erythrocytes). The ruffles are due to a process called crenation, which occurs commonly in red blood cells in vitro. -Marie

On Nov 6, 2013, at 7:06 AM, {eikonika-at-otenet.gr} wrote:

}
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} Dear Friends
}
} While viewing an epithelium in SEM I came across numerous spherical objects
} of 10-20 microns size with their surfaces forming ruffles (please click this
} link http://www.eikonika.net/v2/download.php). These ruffles remind me of
} the appearance of mucous I have seen before.
} Can anybody comment on the origin of these objects? Could they be epithelial
} cells undergoing mucinous degeneration?
}
} Thanks in advance to the wise List
}
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} Tel/fax +30 210 8957677
} mobile +30 6945 107477
} www.eikonika.net www.aim.cat
} *************************************
}
}
} ==============================Original Headers==============================
} 7, 22 -- From eikonika-at-otenet.gr Wed Nov 6 05:59:57 2013
} 7, 22 -- Received: from echidna.otenet.gr (smtp-out24.otenet.gr [83.235.69.24])
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} 7, 22 -- From: "yorgos nikas" {eikonika-at-otenet.gr}
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} 7, 22 -- Subject: object to identify in SEM
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Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369

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I'm glad you asked this question because I see these structures *all the
time* on marine tissues or isolates of marine microbes. I don't know
exactly what the precipitate is. I use 4% glutaraldehyde in 0.1M sodium
cacodylate buffer with sucrose to adjust the osmolarity, avoiding
phosphate buffers. It does, indeed, look like hydroxyapatite, although my
chemistry is not good enough to decide if my fixation protocol is causing
this. For a long time we thought it was some sort of microbe, but I'm
reasonably sure it is not. It COULD be mucous from my samples, now that
you mention it...

To those who only see red blood cells, you need to scroll to the right or
enlarge your browser window to see the precipitates.

Aloha from warm and sunny Honolulu,
Tina

} While viewing an epithelium in SEM I came across numerous spherical objects
} of 10-20 microns size with their surfaces forming ruffles (please click this
} link http://www.eikonika.net/v2/download.php). These ruffles remind me of
} the appearance of mucous I have seen before.
} Can anybody comment on the origin of these objects? Could they be epithelial
} cells undergoing mucinous degeneration?
}
} Thanks in advance to the wise List
}
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} Tel/fax +30 210 8957677
} mobile +30 6945 107477
} www.eikonika.net www.aim.cat
} *************************************
}
}
} ==============================Original Headers==============================
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****************************************************************************
* 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*
****************************************************************************

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Sorry for the confusion. I was referring to the normal red blood cells throughout the field, and one apparently crenulated red blood cell in a pocket near the bottom of the field (due north of the scale bar). After rereading the original letter I realized it was referring to the larger crystalline structures. -Marie

Dr. Marie E. Cantino
Associate Professor of Physiology and Neurobiology
Director, Electron Microscopy Laboratory
University of Connecticut, Unit 3242
Phone: 860-486-3588
Fax: 860-486-6369

==============================Original Headers==============================
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At my point of view that structures could be atypical cells, cells
that appear abnormal under a microscope.
I have met them before in a lung epithelium of a chicken embryo.

Panagiotis

Quoting marie.cantino-at-uconn.edu:

} ----------------------------------------------------------------------------
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} The disc/biconcave disc-shaped structures are almost certainly red
} blood cells (erythrocytes). The ruffles are due to a process called
} crenation, which occurs commonly in red blood cells in vitro. -Marie
}
} On Nov 6, 2013, at 7:06 AM, {eikonika-at-otenet.gr} wrote:
}
} }
} }
} }
} } ----------------------------------------------------------------------------
} } The Microscopy ListServer -- CoSponsor: The Microscopy Society of America
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} } Dear Friends
} }
} } While viewing an epithelium in SEM I came across numerous spherical objects
} } of 10-20 microns size with their surfaces forming ruffles (please click this
} } link http://www.eikonika.net/v2/download.php). These ruffles remind me of
} } the appearance of mucous I have seen before.
} } Can anybody comment on the origin of these objects? Could they be epithelial
} } cells undergoing mucinous degeneration?
} }
} } Thanks in advance to the wise List
} }
} }
} } Dr Yorgos Nikas
} } Athens Innovative Microscopy
} } Skra 36 Voula 16673 GREECE
} }
} } Tel/fax +30 210 8957677
} } mobile +30 6945 107477
} } www.eikonika.net www.aim.cat
} } *************************************
} }
} }
} } ==============================Original Headers==============================
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}
} Dr. Marie E. Cantino
} Associate Professor of Physiology and Neurobiology
} Director, Electron Microscopy Laboratory
} University of Connecticut, Unit 3242
} Phone: 860-486-3588
} Fax: 860-486-6369
}
}
}
} ==============================Original Headers==============================
} 6, 19 -- From marie.cantino-at-uconn.edu Wed Nov 6 12:17:55 2013
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--
Dr. Berillis Panagiotis
Lecturer in Microscopy, image analysis and histology
Department of Ichthyology and Aquatic Environment
Agricultural school
University of Thessaly

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I and my colleagues sometimes see needle-shaped crystalline precipitates in SEM preps of marine samples (fish gills, crustacean larvae). Does anyone have a sample prep that might eliminate these various kinds of precipitates? (I do remember hearing long ago that seawater can precipitate on surfaces upon fixation, but I don't remember any details.)
--Best, Jan Factor

Jan Robert Factor, Ph.D.
Professor of Biology, Chair of Biology Program
Purchase College, State University of New York
Purchase, NY 10577
Office: 2016 Natural Science Bldg., 914-251-6659
Office Hours (Fall 2013): Mon., 10:30-12:00, and Wed., 4:30-6:00
jan.factor-at-purchase.edu

Purchase College ranked in the Top 10 public colleges nationally
Please consider the environment before printing this e-mail

________________________________________
X-from: tina-at-pbrc.hawaii.edu {tina-at-pbrc.hawaii.edu}
Sent: Wednesday, November 6, 2013 2:30 PM
To: Factor, Jan

I'm glad you asked this question because I see these structures *all the
time* on marine tissues or isolates of marine microbes. I don't know
exactly what the precipitate is. I use 4% glutaraldehyde in 0.1M sodium
cacodylate buffer with sucrose to adjust the osmolarity, avoiding
phosphate buffers. It does, indeed, look like hydroxyapatite, although my
chemistry is not good enough to decide if my fixation protocol is causing
this. For a long time we thought it was some sort of microbe, but I'm
reasonably sure it is not. It COULD be mucous from my samples, now that
you mention it...

To those who only see red blood cells, you need to scroll to the right or
enlarge your browser window to see the precipitates.

Aloha from warm and sunny Honolulu,
Tina

} While viewing an epithelium in SEM I came across numerous spherical objects
} of 10-20 microns size with their surfaces forming ruffles (please click this
} link http://www.eikonika.net/v2/download.php). These ruffles remind me of
} the appearance of mucous I have seen before.
} Can anybody comment on the origin of these objects? Could they be epithelial
} cells undergoing mucinous degeneration?
}
} Thanks in advance to the wise List
}
}
} Dr Yorgos Nikas
} Athens Innovative Microscopy
} Skra 36 Voula 16673 GREECE
}
} Tel/fax +30 210 8957677
} mobile +30 6945 107477
} www.eikonika.net www.aim.cat
} *************************************
}
}
} ==============================Original Headers==============================
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****************************************************************************
* 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*
****************************************************************************

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realname - Iftikhar
Email - ahmad-at-cec.sc.edu
ORGANIZATION - University of South Caroloina
EDUCATION - Graduate College
LOCATION - Columbia, SC, USA
SUBJECT_OF_QUESTION - Is ther any third party to Repair FESEM
QUESTION - Hello,

Can you guide me to some person or company which can repair Zeiss FESEM.

Thanks,

Iftikhar

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Dear All

Thanks to You I got many responses regarding this unidentified sem
object -maybe we should name it "uso", reminiscent of a local spirit called
"ouzo". I found all responses very appealing (pity that some were sent off
list) and since I see a broader interest about this "uso" I would like to
summarize the responses and to share with you some thoughts.

When you see biology samples everything is passing from your mind, so I was
particularly thrilled by the possibility that this object may be an
activated leukocyte. However, as pointed out by Phil, the ruffles are too
sharp and also, the sizes I've seen vary from 3 to 25 microns making it
unlikely to be such cell.

Another interesting thought is that the object may be a microbes. The shape
reminds an ureaplasma colony, yet the latter is more like a sea-urchin and
there were no other signs of infection on the specimen (that is human
endometrium).

A third possibility is that "uso" is an epithelial cell undergoing some kind
of change. Mucinous cell degeneration was initially in my mind because I've
notice these objects come out more frequently in hyperestrogenized
endometria that have abundant mucus. Also, Panagiotis reported similar
objects on lung epithelium. However, I see them not only on the epithelium
but also in the stroma. And after Your many responses pointing to phosphate
crystals I am inclined to believe -to my disappointment- that the "uso" is
just a soulless crystal that should be rejected as "artifact". Indeed my
fixation medium includes phosphates (PBS). Tina and other investigators
report crystals without using phosphates, but of course they may be of
different composition.

Here I would like to show you 3 images that may tell us something about the
nature of crystallization in biology samples:

In this photo
http://www.eikonika.net/v2/download1.php
we see endometrial epithelium on the right (PLEASE SCROLL!!!) followed by
naked stroma and in the middle we see many objects residing deep inside an
endometrial fold, along with many RBC. This location is very unlikely to
reach from outside, so these objects must have been grown in situ.

In the second photo
http://www.eikonika.net/v2/download2.php
we see that the objects tend to co-localize with RBC and degenerate cells.
Also, these objects tend to conglomerate.

In the third photo
http://www.eikonika.net/v2/download3.php
we see probably the beginning of the object formation where these "ruffles"
seem to arise from a red blood cell.

I think these objects are phosphate crystals nucleating from red blood cells
or from other cells that may be special in some way: could be degenerate or
have some changes on their coating, including the surrounding mucus.
Thus the presence of these crystals may have some biological significance.

I'll send a sample to Stefan to analyze it with EDS and let you know about
the results
Have a nice day
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************

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Name: James Weston

Organization: NYU Abu Dhabi

Title-Subject: [Filtered] How do you mount algal cells before fixation, fluid replacement, and
critical point drying?

Message: Hello -
Long time listener, first time caller....
I am a materials engineer and I am assisting an external researcher in preparing algal samples (~10
um) for SEM imaging. We have a recipe for fixation, fluid replacement, CP drying and coating, but my
concern is this.

If we go through the procedure and get to the critical point drying, how do we mount the cells on a
stub prior to drying, so that they are not washed out of the alcohol solution when the CO2 evaporates?

Assistance greatly appreciated.
James

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James hi

First of all you must have a suspension of cells. You can filter your
cells through a millipore filter. Your cell will remain onto the
filter (supposing that the pore size is smaller than the cell size).
Then you can proceed dehydration easily by dropping alcohol onto the
filter. The cells will go nowhere. To proceed critical point dry you
can put an other empty filter on the filter that contains the cells.
You can put some glue to the edges of the one filter and let the other
filter to glue on it. Then you can put the "sandwich" TO THE CRITICAL
POINT DRYER.

Panagiotis

Quoting microscopylistserver-noreply-at-microscopy.com:

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} Title-Subject: [Filtered] How do you mount algal cells before
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} critical point drying?
}
} Message: Hello -
} Long time listener, first time caller....
} I am a materials engineer and I am assisting an external researcher
} in preparing algal samples (~10
} um) for SEM imaging. We have a recipe for fixation, fluid
} replacement, CP drying and coating, but my
} concern is this.
}
} If we go through the procedure and get to the critical point drying,
} how do we mount the cells on a
} stub prior to drying, so that they are not washed out of the alcohol
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} James
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--
Dr. Berillis Panagiotis
Lecturer in Microscopy, image analysis and histology
Department of Ichthyology and Aquatic Environment
Agricultural school
University of Thessaly

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Dear Yorgos,

Can you look at your samples after each processing stage to see when these
structures appear? They look similar to many types of crystal (mostly
calcium oxalate) seen in certain plant cells- check out this cover image -
http://pcp.oxfordjournals.org/content/53/7.cover-expansion - the calcium
oxalate crystal is quite similar to your images. They do come in a variety
of shapes and sizes.
Are bladder crystals calcium oxalate?

My 2c,
cheers,
Rosemary

Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
M 61 2 420 972 028
E rosemary.white-at-csiro.au

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Hello James,
We quite often work with cells in suspension. We have a protocol which works
splendidly! We use the 12mm round coverslips which fit nicely on our stubs.
The coverslips are flooded with 0.1% poly-l-lysine and left in a dust free
environment for an hour. After wicking off the poly-l-lysine, the prepared
coverslip is flooded with the sample and left in a dust free environment for
an hour. Remember to keep the solution on the coverslip and don't let it
spill over the sides. After an hour, the excess solution is wicked away. The
coverslips are put into a special holder made specifically to hold them for
critical point drying. We purchased it from Tousimis (I've also used a small
spring to hold the coverslips which is a bit more difficult). Once the
coverslips are in the holder, they are dehydrated and critical point dried
easily. The finished samples are mounted onto our stubs with double sticky
carbon tabs and then sputter coated.
If you need our protocol, please let me know off line and I can send it to
you.
Good Luck!

Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

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Email: james.weston-at-gmail.com
Name: James Weston

Organization: NYU Abu Dhabi

Title-Subject: [Filtered] How do you mount algal cells before fixation,
fluid replacement, and critical point drying?

Message: Hello -
Long time listener, first time caller....
I am a materials engineer and I am assisting an external researcher in
preparing algal samples (~10
um) for SEM imaging. We have a recipe for fixation, fluid replacement, CP
drying and coating, but my concern is this.

If we go through the procedure and get to the critical point drying, how do
we mount the cells on a stub prior to drying, so that they are not washed
out of the alcohol solution when the CO2 evaporates?

Assistance greatly appreciated.
James

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James,

I'm not sure what you mean by "...washed out of the alcohol solution
when the CO2 evaporates?" There should not be any alcohol in the CPD
when the actual drying run is made - otherwise you'll be drying from
alcohol, and the cells will collapse.

The best methods we've found are:
1) Collect the algal cells onto a membrane filter - the type with holes,
not a torturous-path filter - mounted in a syringe filter holder or a
filter column. Do your fixation and dehydration series in this apparatus
as well.
After the final alcohol wash, transfer the membranes with cells into the
CPD device (use a basket or something to contain the filter membranes),
do the flush/soak cycles to remove all the alcohol from within the cells
and replace with lqCO2. Then make the run.

2) Instead of CPD, use HMDS (hexamethyldisilizane) **in a fume hood!**.
This will take a bit of testing to find the right conditions for your
cells, but: still using the syringe or filter column, replace the
alcohol with HMDS, using a transition series (2:1 alcohol:HMDS, 1:2 or
add a 1:1 step), then 3 washes with HMDS. In or after the 3rd wash,
place the filter in watch glass or similar with just enough HMDS to
cover the filters, and cover the dish with a cocked lid or lint-free
paper, enough to prevent dust etc. from falling on the samples, but
leave space for evaporation.
Algal cells will usually dry best at 60 deg C, 2-4 hours (the oven must
be in a fume hood). Room temperature can also work - this is one of the
things to test.

There is a potential 3rd method being worked on by Tobias Baskin that
may well work for algae - I haven't tried it yet, myself.

Phil

} Email: james.weston-at-gmail.com
} Name: James Weston
}
} Organization: NYU Abu Dhabi
}
} Title-Subject: [Filtered] How do you mount algal cells before fixation, fluid replacement, and
} critical point drying?
}
} Message: Hello -
} Long time listener, first time caller....
} I am a materials engineer and I am assisting an external researcher in preparing algal samples (~10
} um) for SEM imaging. We have a recipe for fixation, fluid replacement, CP drying and coating, but my
} concern is this.
}
} If we go through the procedure and get to the critical point drying, how do we mount the cells on a
} stub prior to drying, so that they are not washed out of the alcohol solution when the CO2 evaporates?
}
} Assistance greatly appreciated.
} James
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Colleagues

You may have noticed a quiet period or connection
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There was a hard line (fiber optic) failure to my
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The downstream line has finally been repaired and hopefully,
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Anyway, we should be back running again.

Cheers,

Nestor
Your Friendly Neighborhood SysOp

P.S.

Oct 1, 2013, was the 20th anniversary of
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Dear Prof. Dr. Zaluzec,
good morning, dear Nestor,

just to say THANK YOU SO MUCH for all your efforts during the last (20!) years to keep the Listserver running.
I always felt it to be a unfathomable {personalized inverted "black hole"} serving colleagues in need of more/less specific information...really a source of wisdom and collegiality.

20 years!!
� votre sant�,
cheers

respectfully but cordially yours,
Wolfgang

SALZBURG, AUSTRIA

Von: zaluzec-at-aaem.amc.anl.gov [mailto:zaluzec-at-aaem.amc.anl.gov]
Gesendet: Freitag, 08. November 2013 20:26
An: Mu� Wolfgang
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Colleagues

You may have noticed a quiet period or connection problems during the last few days on the Microscopy Listserver.

There was a hard line (fiber optic) failure to my servers. Although the servers were running locally, I've
had no connection/links to the outside world....

The downstream line has finally been repaired and hopefully, the new cabling that was installed somewhere
a few miles from here will fix the major issue.
I wouldn't hold my breath, as there is likely another culprit hiding in the wings that created the original fault, so don't be surprized if we go quiet again for abit, my local last-mile Tel-Co is continuing their testing and rerouting of traffic to identify the cause of the fault.

This has been quiet literally the longest outage I've ever had here in my quiet little neck of the woods.
It is amazing how one takes connectivity and access for granted today.

Anyway, we should be back running again.

Cheers,

Nestor
Your Friendly Neighborhood SysOp

P.S.

Oct 1, 2013, was the 20th anniversary of the start of this forum. Everyone feel free to raise your favorite virtual drink glass/mug and congratulate each other for having a LONGER social media history than those more recent
upstarts.

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I would like to see examples, or links, and hear opinions the state the histological (Optical or EM) technology of "normal"(and in various hormonal stages?), micro-biologically pathological, and cancer breast tissue? Since this technology seems generally highly automated, I think most Histology and R&D professionals (all levels) would assume a high general confidence level, but it seems with automation, compromise and range of conditional restrictions, there might be a level of necessary compromise present. Are there any? What is your opinion?

- Secondarily, I am also interested in opinions about issues concerning 'post-mortum and biopsy'(pre-fixation pre-and post-fixation timing and sample collection technique issues ) histology of various levels and conditions of fixation (cryo- and RT). I think this is the historical basis of "Historical Histo-Pathology". Finally two more issues, the histological interpretation of the effects of : 1) thick and thin section issues and 2) serial section image analysis/path and normal issues (I seem to have seen these issues more frequently discussed among skin cancer histology technique studies).

Since the scope of this inquiry is very broad, I would prefer current, historical, and text references, links and some limited discussion, please, here or directly to my email, wa5ekh-at-juno.com

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Email: gul417-at-mail.usask.ca
Name: Guosheng Liu

Organization: U of Saskatchewan

Title-Subject: [Filtered] Philips CM10 TEM specimen translation control
stuck

Message: Dear All,

The Y-translation control on our CM10 TEM stuck last week during
"normal" operation by an "experienced" user. The symptoms include
1)stiff Y-control / the light on the control was ON; 2) goniometer
(specimen holder entry part) stuck at the left side and immobile (see
photos at
file:///W:/TEM%20specimen%20Y-translation%20control.html).

I removed the cover of tilt mechanism and tried to find any things wrong
mechanically. I also restarted the machine but no success. This happened
once a few year ago and a service call was made. Due to the lack of
service contract right now, we are wondering if there is a "simple" way
to fix it by the guidance of experts out in the forum.

Any tips/suggestions would be greatly appreciated.

Best regards.

George Liu
U of S, Biology
Canada

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George,

We have a CM-10 and frequently get a jammed right-side translation rod.
(Y-drive, although the X/Y is confusing to folks, since the image rarely
moves in the X or Y direction.)

Question: Motor drive (foot pedals) or strictly hand-cranked?
Can you remove and insert the specimen rod normally?

A sticky Y-drive is common. Have you tried grabbing the translation rod
up next to the airlock (above all the joints) and turning it? That
usually works for us. Stuck all the way left should mean turning the top
of the rod away from the column.

Is the rod jammed with the goniometer motor both engaged and disengaged?
And can you manually tilt the stage with the motor disengaged? If yes,
then it is strictly a translation rod problem.

Is the Z-drive centered, or at one or another end of its range of
motion? If not centered, try that.

Can you push on the end of the specimen rod, and move the rod laterally
that way? Mimicking the action of the translation rod by pushing
sideways on the rod-end cap. If the rod moves freely, the problem again
is most likely the translation rod, or where it engages the drive
gearing to move the rod. If the rod doesn't move, it's jammed in the
airlock, and there may be nothing wrong with the translation rod.

If it's the latter, or the drive gearing in the airlock is jammed, or
you cannot manually tilt the stage, you'll most likely have to pull the
airlock off the column.

First, though, hope for the best: the rod moves freely sideways, so the
rod drive is jammed. But the rod can be turned at the uppermost end,
where it joins the airlock. Push the rod to the middle and turn the rod.
Carefully.
This is a common jam, and gets frozen hard when a user first hits the
jam and then keeps trying to move the specimen, tightening the jam.
Careful work can still free it: if rod end is all the way left, turn the
top of the rod away from the column; if all the way right, turn the top
of the rod toward the column. If jammed hard enough, you may need a pair
of pliers with padded jaws to get it started.
BUT be careful! Done wrong, that will make things worse and potentially
expensive!

Is the rod stiff down at the handle? Reads like it, but loose set screws
on the rods (not the U-joints) will also cause loss of stage movement.
The rod spins freely then, however.

Phil

} Email: gul417-at-mail.usask.ca
} Name: Guosheng Liu
}
} Organization: U of Saskatchewan
}
} Title-Subject: [Filtered] Philips CM10 TEM specimen translation control
} stuck
}
} Message: Dear All,
}
} The Y-translation control on our CM10 TEM stuck last week during
} "normal" operation by an "experienced" user. The symptoms include
} 1)stiff Y-control / the light on the control was ON; 2) goniometer
} (specimen holder entry part) stuck at the left side and immobile (see
} photos at
} file:///W:/TEM%20specimen%20Y-translation%20control.html).
}
} I removed the cover of tilt mechanism and tried to find any things wrong
} mechanically. I also restarted the machine but no success. This happened
} once a few year ago and a service call was made. Due to the lack of
} service contract right now, we are wondering if there is a "simple" way
} to fix it by the guidance of experts out in the forum.
}
} Any tips/suggestions would be greatly appreciated.
}
} Best regards.
}
} George Liu
} U of S, Biology
} Canada
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Name: Ruth Yamawaki

Organization: Stanford University

Title-Subject: [Filtered] Free older Gatan Camera

Message: I inherited a older model Gatan Camera but never installed it.
We are cleaning house and it is time for it to go.

Since I never used it I am not sure exactly what I have or if I have the
whole ensemble. The book says model 689 Wide Angle slow-Scan CCD Camera.

Yours for free but you pay for shipping.

Contact me if you are interested or have further questions.

Best,
Ruth

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Email: michael.cammer-at-med.nyu.edu
Name: Michael Cammer

Title-Subject: [Filtered] coverslip edge question

Message: We looked at the edge of #1.5 coverslips (Item 10812 for Ibidi
sticky slides) and found that they appear cut from two sides. We figure
this is to prevent chipping or shattering during cutting.

Picture at http://www.flickr.com/photos/mcammer/10856843116/ or
http://www.ipernity.com/doc/285585/28173425

Just curious if this is common for coverslips.

Thank you very much.

Michael Cammer

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Email: michael.cammer-at-med.nyu.edu
Name: Michael Cammer

Title-Subject: [Filtered] coverslip edge question

Message: We looked at the edge of #1.5 coverslips (Item 10812 for Ibidi
sticky slides) and found that they appear cut from two sides. We figure
this is to prevent chipping or shattering during cutting.

Picture at http://www.flickr.com/photos/mcammer/10856843116/ or
http://www.ipernity.com/doc/285585/28173425

Just curious if this is common for coverslips.

Thank you very much.

Michael Cammer

==============================Original Headers==============================
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Hello,

we found this hexagonally shaped structure near the nucleus in an
LR-White embedded heart tissue (mouse) for immunolabeling. We presume
that this structure is a virus. Is this correct?

http://chimerism.medunigraz.at/Pictures/26.php

Thank you for your help!
Stefan

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My questions are:
1. Are there really smooth and rough surfaces, and which side should be exposed to the mountant.
2. Re. #1: Are all coverslips in a box stacked rough up, or are they randomly packaged?
3. AND, what might be the optical consequence of ignoring this information?

One consequence is to be ignorant of how our tools/consumables are fabricated.

Thanks,

Fred Monson

Frederick C Monson, PhD
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pannsylvania
West Chester, PA, 1938
610-738-0437
fmonson-at-wcupa.edu
________________________________________
X-from: oshel1pe-at-cmich.edu [oshel1pe-at-cmich.edu]
Sent: Friday, November 15, 2013 8:16 AM
To: Monson, Frederick

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Email: michael.cammer-at-med.nyu.edu
Name: Michael Cammer

Title-Subject: [Filtered] coverslip edge question

Message: We looked at the edge of #1.5 coverslips (Item 10812 for Ibidi
sticky slides) and found that they appear cut from two sides. We figure
this is to prevent chipping or shattering during cutting.

Picture at http://www.flickr.com/photos/mcammer/10856843116/ or
http://www.ipernity.com/doc/285585/28173425

Just curious if this is common for coverslips.

Thank you very much.

Michael Cammer

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Dear Colleagues,

At the age of 74 and in preparation for ........

I have in my possession the following:

1. 1 Liter of Gum Damar in Xylene - perfect consistency for mounting. Probably the best for archival slides, but too much work to make up for automatic use. Personally fabricated in mid 1990's just in time for the beginning of my, now long hiatus from my favorite employment - histology. The original lumps were defatted, so after dissolving in xylene, it was filtered, and transferred periodically to smaller containers after evaporative reduction in handy chemical hoods. Once fitted to a Liter bottle a permanent cap was used to stop the process. The bottle has resided in cool and dark environments since.
2. 1 pint of Canada Balsam in xylene - Fisher.
3. two small jars of quite viscous Canada Balsam (~8 oz each)

If interested, please contact me offline with purpose included.

Cheers,

Fred Monson

Frederick C Monson, PhD
Center for Microanalysis and Imaging, Research and Training (CMIRT)
West Chester University of Pannsylvania
West Chester, PA, 1938
610-738-0437
fmonson-at-wcupa.edu

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Dear Stefan,

I�ve never worked with viruses; but if I saw that in a neuron, I would
probably think it was a dense core vesicle.

The outer structure does appear to be angular, however, and looking at
some images of viruses, it does look like it could be a large virus.

Not much help, sorry!

Ben

--
Research Support Manager
MRC Anatomical Neuropharmacology Unit, Mansfield Road,
Oxford, OX1 3TH, United Kingdom. Telephone: 01865 271649
{http://www.mrc.ox.ac.uk/}

On 15/11/2013 13:33, "stefan.wernitznig-at-medunigraz.at"
{stefan.wernitznig-at-medunigraz.at} wrote:

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15, 31 -- From ben.micklem-at-pharm.ox.ac.uk Fri Nov 15 08:47:25 2013
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stefan.wernitznig-at-medunigraz.at

Dear Stefan,
really pretty nice image you have...(:-))

Not being an expert with specimen processing(?) and embedding in LR-White and perhaps also not in terms of "virology", nevertheless I consider your structure (measured on/from your provided image-document) to be an
icosa(penta-)hedral virus structure, being something like 210-220 nm in ?outer envelope? diameter, diameter of the imaged nucleocapsid/core around 150 nm, which would fit +/less perfectly for Herpesvirus / herpesvirion (enveloped, icosahedral nucleocapsid, diameter 100-300 nm).

Interestingly for me: the "hexagonal" structure you mentioned seems to be ?part of the envelope/tegument? of the virus particle (misinterpretation expected to be corrected...THANK YOU!).
Any herpes-virus image I have in mind (and have found rushing through 3 classical virus-EM-books) looks different from another [and therefore - for ultrathin sections of such viruses - ONE typical morphological feature , it seems, does not exist] and also in several images there are shown ROUND(ish) outer envelopes as well as nucleocapsid circumferences too.

Unfortunately you did not say where the specimen comes from (only mouse heart tissue), but I guess it was "real" mouse heart from the animal, not cultured cells.
So you are lucky to have gotten such a nice image.
Herpes Viruses in heart tissue are believed to exist... cf (eg.)

-at-http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3547968/�
PLoS One.�2013;�8(1): e54008. Published online 2013 January 17.�doi:��10.1371/journal.pone.0054008
PMCID:�PMC3547968
{Evidence for the Role of Epstein Barr Virus Infections in the Pathogenesis of Acute Coronary Events}
by Philip F. Binkley et al.
"PLoS One published in January, 2013 states that infection with latent Epstein Barr Virus (EBV) may lead to heart disease and heart attacks"

-at-http://www.ncbi.nlm.nih.gov/pubmed/9236842 or:
-at-http://journals.lww.com/ajsp/pages/articleviewer.aspx?year=1997&issue=07000&article=00014&type=abstract
American Journal of Surgical Pathology:
July 1997 - Volume 21 - Issue 7 - pp 847-853
{Herpesvirus 6 Variant A Infection After Heart Transplantation with Giant Cell Transformation in Bile Ductular and Gastroduodenal Epithelium}
Randhawa,�et al.
or.
-at-http://www.health-matrix.net/2013/08/06/heart-attacks-cfs-herpes-virus-infection-and-the-vagus-nerve/
{Heart attacks, CFS, herpes virus infection and the vagus nerve}
Med Hypotheses. 2013 Jun 18. Chronic fatigue syndrome from vagus nerve infection: A psychoneuroimmunological hypothesis. Vanelzakker MB.
and, last but not least:
-at- http://www.ncbi.nlm.nih.gov/books/NBK8157/
"Herpesviruses have a unique four-layered structure: a core containing the large, double-stranded DNA genome is enclosed by an icosapentahedral capsid which is composed of capsomers. The capsid is surrounded by an amorphous protein coat called the tegument. It is encased in a glycoprotein-bearing lipid bilayer envelope..."

It might be that you perhaps are observing a Herpes 6 or (latent) EBV or CMV virus particle (depending on the physiological state or circumstances of the processed tissue).

Hoping there will also some post replies from the virus-experts...,(:-))
Best wishes and regards,

ALL: have a beautiful and recreative weekend,

Wolfgang MUSS
EM-Lab, Univ. Inst. Pathol.
SALK-LKH (Gen. Hospital) & PMU (Paracelsus Medical University)
SALZBURG, Austria
==============================================================================================

Von: stefan.wernitznig-at-medunigraz.at [mailto:stefan.wernitznig-at-medunigraz.at]
Gesendet: Freitag, 15. November 2013 14:31
An: Mu� Wolfgang
Betreff: [Microscopy] hexagonally shaped structure

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we found this hexagonally shaped structure near the nucleus in an
LR-White embedded heart tissue (mouse) for immunolabeling.
We presume
that this structure is a virus. Is this correct?

http://chimerism.medunigraz.at/Pictures/26.php

Thank you for your help!
Stefan

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Iftikhar,
I haven't seen any replies on the server so it's a little hard to tell if
you have had any replies off line.

Unless you are some extremely rare person who has gotten their hands on the
schematics/service manual for a Zeiss SEM, I think you are going to be
restricted to using Zeiss, which is exactly what they had in mind. Best of
luck.

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

-----Original Message-----
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Sent: Wednesday, November 06, 2013 3:12 PM
To: kenconverse-at-qualityimages.biz

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Forwarded from "Ask a Microscopist"
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realname - Iftikhar
Email - ahmad-at-cec.sc.edu
ORGANIZATION - University of South Caroloina
EDUCATION - Graduate College
LOCATION - Columbia, SC, USA
SUBJECT_OF_QUESTION - Is ther any third party to Repair FESEM
QUESTION - Hello,

Can you guide me to some person or company which can repair Zeiss FESEM.

Thanks,

Iftikhar

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Josh, you can not buy the "full package". You may however buy individual
sizes as needed. Available cheaply from industrial suppliers, less so
from dedicated vacuum parts/materials suppliers. Same material and same
size seal does same thing regardless of a price tag.

Know the size of the O-ring. Could be inch (fractional or dash number)
or metric (actual mm). Interchangeable or at least close enough in most
cases.

Available from multiple suppliers. I like www.mcmaster.com . Look for
section "Sealing" and then the sub-section "O-rings". Other suppliers
have same large selection of sizes and materials.

Do not use Buna for EM applications. Use Viton instead.

CM-10 has very few special seals (other than O-rings) available from FEI
or Edwards Vacuum - see www.edwardsvacuum.com . But O-rings are standard
and nothing special.

Vitaly Feingold
SIA
2773 Heath Lane
Duluth GA 30096
Ph. 770-232-7785
Fax 770-232-1791
www.sia-cam.com
vitaly-at-sia-cam.com

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Title-Subject: [Filtered] Looking for CM10/CM12

Message: We are looking for someone that is wanting to get rid of/donate
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Email: escanley-at-gmail.com
Name: Ellen Scanley

Organization: Southern Connecticut State University

Title-Subject: [Filtered] Scanning Transmission Electron Microscopy

Message: Hi Microscopy ListServe,

I am trying to learn about STEM. It seems
like one of the really important things is
that the electrons reaching the HAADF
detector are incoherent.
I am assuming that the HAADF detector is
recording a net amplitude (?intensity?) for
each raster position (i.e. an image is not
formed on the HAADF detector). Why does the
incoherence increase resolution?

The diagrams I have seen show two point sources
and the different convolution with the point
spread function for incoherent and coherent
illumination. That seems to make sense for an
image to distinguishing the two points but I don't
follow how that relates to the integrated signal
over the HAADF detector.

Many Thanks,
Best Regards,
Ellen

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Dear Microscopy-Listers,

thank you for all the informative answers!

At first I have to say, that the picture is not from me, so all the
credit goes to one of my colleagues - so if the structure would be
something totally new, it would be a "Lissi Body"! [Pat] ;)

This picture was the best one of the structure, but I will ask if there
are other ones that I can post, maybe an overview in low mag.

We will go through all the infos you gave us and I will come back to you
later this week!

Best wishes,
Stefan

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Name: Richard Gursky

Organization: St. Jude Childrens Research Hospital

Title-Subject: [Filtered] Edwards Carbon Rod Evaporation Kit

Message: Hello,

We are looking for an Edwards Carbon Rod Evaporation Kit. We need just
the parts that you see in the Bell Jar, (but we need all of them) the
Stand Rods, Braided Cu Cable and brackets & the 2 parts of the Carbon
Rod Clamping / Evaporation device.

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Title-Subject: [Filtered] Changing filament on CM10

Message: Can someone aid me in replacing the filament in the Philips
CM10? I have the Wehnelt Cylinder exposed, but it looks like you need a
special tool to remove the notched "bolt" at the bottom. If this is the
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Hi Josh,

You shouldn't need any special tools to change a filament in a
Philips/FEI wehnelt. The cap is held on with a spring clip and you
should be able to pull it off. FEI does have some thin levers to help
pry it off if it is really tight, but generally you shouldn't need them.

The "notched" bolt is the inner assembly which is used to set the
filament depth. When you remove the cap, you will see a small locating
pin on the inside. After replacing the cathode and cleaning the inside
of the wehnelt aperture, you can turn the cap and it will adjust the
filament depth (generally ~200-220um).

The owners manual has a good description of the process.

Cheers,
Henk

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}
} Message: Can someone aid me in replacing the filament in the Philips
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} special tool to remove the notched "bolt" at the bottom. If this is the
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"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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Email: researchers4u-at-yahoo.com
Name: sanaz shobeikeh

Organization: shiraz university

Title-Subject: [Filtered] cm-10 philips transmission electron microscopy
Maintenance

Message: hi :
I'm a new technician of shiraz university's TEM lab(country: iran). we
have a CM-10 PHILIPS and because of university's budget problems can't
hire maintenance professional. could anyone please guide me through
filament changing , odp and rotary pump oil changing , aperture and
condenser cleaning and other important Maintenance related works ? also
recently when TEM is working for about an hour , it's high tension will
automatically turns off (green light go off but in screen is still on
100kv) and afterwords when i try to turn the filament back on with it's
volume(about step 14): emission meter pointer shows maximum number
(before the pointer showed a value between 20 to 25). i have the cm-10
operational manual , but it's my first time and i'm really afraid of
making a mistake , please guide me through this situation. thanks for
your time.

best regards

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Hi Sanaz,
first question:
Do you have all the manuals for the CM-10? If yes, there should be a lot of procedures inside for solving your problems.

If not, there are some things I would like to know:
- Cathode is Tungsten or LaB6?
- What is the end-pressure you are reaching now after pumping for a day?
- When you switch on high voltage (without heating the cathode) does the value go down to nearly zero or is it somewhat higher or
even max of the scale?
- Do you have dry nitrogen gas for flushing the column during exchange of wehnelt / cathode assembly?

Concerning change of pump fluids:
- If you have trouble with reaching the vacuum values, this might also indicate a leak or an old ion getter pump.
I would not change the Santovac fluid of the diffusion pump, normally it lasts a lifetime. You also need at least one special
o-ring to do so.
- Changing the pre-vac pump fluid: Let the pump go warm, open up the screw at the bottom of the pump and drain the old old. Put in
30% of needed oil for one filling; let the pump work for 15 seconds, drain again. Fill pump with new oil nearly to the maximum
indicated on the pump. Disconnect from microscope system, close the inlet of the pump and let it run on ballast for half an hour
to outgas the oil, then connect it to the microscope again and start pumping.

There should be a detailed procedure for this, when you are searching for the manual at the pump manufacturer`s website...

If you need a short and also a detailed user manual for the CM series: I have one in PDF, but only for the CM-12. But the basics
are the same, like cleaning apertures / wehnelts etc. Ask me off-line, please.

Hope it helps,
Stefan

-----------------------------------------------------
Stefan Diller - Scientific Photography
Arndtstrasse 22
D - 97072 Wuerzburg Germany
++49-931-7848700 Phone
++49-931-7848701 Fax
++49-175-7177051 Mobile

Websites:
www.nanoflight.info
www.stefan-diller.com
www.electronmicroscopy.info
www.elektronenmikroskopie.info
www.zwillingsprojekt.de
www.assisi.de
Anfahrt: http://Mail.map24.com/Stefan.Diller
-----------------------------------------------------

Am 20.11.13 09:33, schrieb microscopylistserver-noreply-at-microscopy.com:
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}
} Title-Subject: [Filtered] cm-10 philips transmission electron microscopy
} Maintenance
}
} Message: hi :
} I'm a new technician of shiraz university's TEM lab(country: iran). we
} have a CM-10 PHILIPS and because of university's budget problems can't
} hire maintenance professional. could anyone please guide me through
} filament changing , odp and rotary pump oil changing , aperture and
} condenser cleaning and other important Maintenance related works ? also
} recently when TEM is working for about an hour , it's high tension will
} automatically turns off (green light go off but in screen is still on
} 100kv) and afterwords when i try to turn the filament back on with it's
} volume(about step 14): emission meter pointer shows maximum number
} (before the pointer showed a value between 20 to 25). i have the cm-10
} operational manual , but it's my first time and i'm really afraid of
} making a mistake , please guide me through this situation. thanks for
} your time.
}
} best regards
}
} Login Host: 79.127.42.149
} Listserver Email Form V - 20120416
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Title-Subject: [Filtered] Offer: LKB-Huxley Ultramicrotome Manual

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Dear All,

I am interested in accelerating preparation processes for light
microscopy and transmission electron microscopy and maybe also
immunolabing. Therefore, I am thinking about buying a lab microwave. I
found 3 on the market, the PELCO Bio Wave, the EMS-820 and and the
EMS-9000 lab microwave oven. Who has any experience with these or
maybe other lab microwaves? As they are quite expensive, I wonder if
they are worth the money?

Best regards,
Anne

Dr. Anne Heller
AG Elektronenmikroskopie
Institut für Botanik (210)
Universität Hohenheim
Postfach, 70593 Stuttgart
Tel. 0711-45922180
Fax. 0711-45923355

==============================Original Headers==============================
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Hi everyone!

At first I must apologize for the little confusion my last comment
caused! With "Lissi Body" I was just refering to a comment from Patricia
Connelly [Maybe a "Stephan Body"], because my colleague Elisabeth (we
call her in short "Lissi") has actually found this structure. So that
was just a fun comment. I am sorry for all that.

I will talk to Elisabeth today about some additional TEM-pictures and so
that I can provide you with some more informations about this structure.

Best wishes,
Stefan

==============================Original Headers==============================
6, 30 -- From stefan.wernitznig-at-medunigraz.at Thu Nov 21 06:31:45 2013
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I have a protocol for quantification of viral particles by negative stain
that uses a Beckman Airfuge with EM-90 rotor.
My question, how essential is it to use the Airfuge? And is there an
alternative method if I don't have access to the Airfuge with EM90 rotor?

Thank you
Karen Kelley
UF

==============================Original Headers==============================
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Hello Anne,
We have been using a PELCO Bio Wave for about 12 years. We have been through
3 generations of the unit and loved every one of them. The model we use now
is programmable so we can set our times, wattage, vacuum etc, store them
then select the protocol we need. I have no experience with the EMS units
but I'm sure they are comparable.
Our clinical sample turnaround time is 2-4 days-fixation to images on the
TEM! Processing the samples takes us only a total of 4 hours. Infiltration
takes a total of 12 minutes microwave time. In regards to immunolabeling, I
process the samples using the microwave (again, processed in 4-6 hours
depending on the type of sample). However, we haven't used it for the
labeling process. We embed the immuno tissue into LR White and polymerize in
the microwave under water. I must say that cutting these blocks is 100%
better when polymerized in the microwave. LR White can be brittle and the
protocol we use gives us blocks that are much easier to cut!
Hope that helps,
Cheers,
Pat Kysar, C.E.M.T.
Chair, Certification Board, Microscopy Society of America
University of California, Davis
School of Medicine
Dept of Pathology and Laboratory Medicine
Electron Microscopy Lab

-----Original Message-----
X-from: Anne.Heller-at-uni-hohenheim.de [mailto:Anne.Heller-at-uni-hohenheim.de]
Sent: Thursday, November 21, 2013 1:13 AM
To: pekysar-at-ucdavis.edu

Dear All,

I am interested in accelerating preparation processes for light microscopy
and transmission electron microscopy and maybe also immunolabing. Therefore,
I am thinking about buying a lab microwave. I found 3 on the market, the
PELCO Bio Wave, the EMS-820 and and the
EMS-9000 lab microwave oven. Who has any experience with these or maybe
other lab microwaves? As they are quite expensive, I wonder if they are
worth the money?

Best regards,
Anne

Dr. Anne Heller
AG Elektronenmikroskopie
Institut für Botanik (210)
Universität Hohenheim
Postfach, 70593 Stuttgart
Tel. 0711-45922180
Fax. 0711-45923355

==============================Original Headers==============================
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14, 28 -- From pekysar-at-ucdavis.edu Thu Nov 21 09:58:14 2013
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Dear All,
Electron microscopy facility team at Institute of Science and Technology Austria provides researchers with well-maintained cutting-edge sample preparation and imaging instrumentation and modern infrastructure.

We are enlarging our team and are searching for:

Technician - Electron Microscopy (f/m) (Full time)

Main Tasks:
� Biological sample preparation for electron microscopy.
� Support and training of users in the field of sample preparation techniques, microscopy imaging and image analysis.
� Care of the Electron Microscopy facility equipment - conduct operational maintenance of sample preparation equipment and alignment of electron microscopes.

Profile of candidate:
� Experience with biological sample preparation for electron microscopy observation.
� Work experience in the field of electron microscopy.
� Work experience in a research environment in the field of natural sciences (biology, chemistry).
� Ability to work independently and team-oriented.
� High degree of reliability, organizational and interpersonal skills.
� Very good knowledge in English.

We welcome flexible and reliable team players who are interested in working in an international environment within a diverse and dynamic working atmosphere.

IST Austria values diversity and is committed to equality.

To apply for this position, please send your application in one combined pdf (including CV, photo, certificates and references) by e-mail to: ludek.lovicar-at-ist.ac.at

Please feel free to forward our job offer to anybody you feel it might be relevant.

Thank you!
Best regards,
Ludek Lovicar

=================================
Ludek Lovicar
Electron Microscopy Facility
Scientific Services Unit
Institute of Science and Technology Austria Am Campus 1
A-3400 Klosterneuburg
Austria

Phone: +43-(0)2243 9000 1066
Mobile: +43-(0)664 604 841 066
Fax: +43-(0)2243 9000-2000
Email: ludek.lovicar-at-ist.ac.at

Visit our website: www.ist.ac.at

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-------- Original Message --------

Josh,

From the FEI/Philips manual:
Water temperature IN: min. 6 deg C, max 20 deg C
Water temperature OUT: 20 deg C +/- 2 deg C

water flow max: 2.4 L/min, nominal 2.1 L/min
rise in water temperature through microscope: 6 deg C at max load and
max flow of 2.4 L/min

Phil

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} Date: Fri, 22 Nov 2013 12:08:36 -0600
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} Email: jcsmtf-at-mail.missouri.edu
} Name: Josh Schorp
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} Organization: University of Missouri
}
} Title-Subject: [Filtered] Water temperature for CM10
}
} Message: What is the proper water temperature to cool the CM10 TEM?
}
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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5, 29 -- From oshel1pe-at-cmich.edu Mon Nov 25 08:06:38 2013
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I would rather use higher end of recommended temperatures - too cold water can result in condensation and corrosion.

Vladimir M. Dusevich, Ph.D.
Electron Microscope Lab Director
(816) 235-2072
371 School of Dentistry
650 E. 25th Street
Kansas City, MO 64108-2784

-----Original Message-----
X-from: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Sent: Monday, November 25, 2013 8:07 AM
To: Dusevich, Vladimir

Josh,

From the FEI/Philips manual:
Water temperature IN: min. 6 deg C, max 20 deg C Water temperature OUT: 20 deg C +/- 2 deg C

water flow max: 2.4 L/min, nominal 2.1 L/min rise in water temperature through microscope: 6 deg C at max load and max flow of 2.4 L/min

Phil

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} Email: jcsmtf-at-mail.missouri.edu
} Name: Josh Schorp
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} Organization: University of Missouri
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} Title-Subject: [Filtered] Water temperature for CM10
}
} Message: What is the proper water temperature to cool the CM10 TEM?
}
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Email: roger.ristau-at- uconn.edu
Name: Roger Ristau

Organization: Univ of Connecticut

Title-Subject: [Filtered] Water temperature for CM10

Message: The aim of water cooling of electron microscope lenses is that the temperature inside the
column be the same as that outside the column. Therefore, the chiller should be set to "room
temperature."

Cheers
Roger

Roger Ristau
Institute of Materials Science
University of Connecticut, Storrs

-------- Original Message --------

Hi Josh

You are being given good advice but perhaps an explanation is required?

The water temperature specified for an instrument in most cases is overkill!
Consider the manufacturer's situation, are we cooling 80kV and 20,000x, or
120kV and 500,000x, two totally different situations. The harder the
instrument is run the more heat is generated within the lenses, the result
of higher currents passing through the lens coils. But what happens when
people switch off the electronics, no heat, so excessive cooling, and
inevitably condensation; disaster!

Instruments working at 200 or 300kV will certainly be left on 24/7 so
condensation is not a problem. But in laboratories that use the instruments
irregularly, over cooling may be a route to big problems. If the instrument
is to be left for long periods, with the electronics switched off, or in a
stand-by mode, the water temperature should be adjusted to be no lower than
a couple of degrees below room temperature. But, with a note on the
instrument, so that a user would be aware of the adjustment and be able to
return to "normal" values.

Over cooling of a power transistor board, or the lenses themselves, may lead
to numerous problems. The most catastrophic problem being objective lens
astigmatism that is beyond correction, due to corrosion of the walls of the
lens block. This does not happen in the short term, but with 15 years plus
of over-cooling and I am sad to say I once had to condemn an instrument.
The lens was beyond repair.

Check your instrument out, place your hand on the objective lens and it
should feel very slightly warm after a full day's work. If it feels cold
then in my mind the water cooling is too much!

Good luck, build a true "feel" for your instrument, none of us like being
too cold!

Steve

Steve Chapman FRMS
Protrain for Consultancy and Courses in Electron Microscopy
Mobile +44 (0) 7711 606967 Fax +44 (0)1280 814007
www.emcourses.com

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-----Original Message-----
X-from: DusevichV-at-umkc.edu [mailto:DusevichV-at-umkc.edu]
Sent: 25 November 2013 21:41
To: protrain-at-emcourses.com

Dear Friends,

One of our students made some polymer sheets (polysulphone). She wants to microtome them for light microscopy. The sheets are so thin and flexible, would be easier to handle by resin embedding prior to microtoming. The Agar low viscosity resin we have requires curing at 60 degree C. She doesn’t like anything over 40 degree C. In addition the material is water sensitive.

Any other resins to suggest please?

Zhou
Dr Z Zhou
Loughborough Materials Characterisation Centre (LMCC)
Department of Materials
Loughborough University
Leicestershire
LE11 3TU
UK

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Hi,
I have used a mixture of methyl and butyl
metacrylates to embed samples for light
microscopy. This requires uv polymerization, and
a little bit of trial and error (with the light
source geometry) to get bubble free embedments.
But the stuff sections beautifully. If your
sample is non-aqueous should be able to
infiltrate directly.

If this sounds promising, let me know and
I can send you a protocol off-line.

Good luck,
Tobias

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Title-Subject: [Filtered] second-hand TEM and SEM

Message: Dear All,

We are looking for second-hand 200 kV Transmission electron microscope
and field emission gun scanning electron microscope with high quality
at low kV.

Many thanks,

Ding

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Email: ahmad_ashkaibi-at-hotmail.com
Name: Ahmad Ashkaibi

Organization: Al-Balqa University - Jordan

Title-Subject: [Filtered] Substitute for mounting wax

Message: Hello everybody,
I need a temporary wax to mount an aluminum sample to a manual grinders
platen, but I cannot get any now. Can I use any commercial super glue
instead?
I am preparing a sample for an FEI TECNAI T12 Transmission electron
microscope.

Thank you.
Ahmad Ashkaibi

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Hi Everyone!

Are you aware of a company or group that can provide some nanocubes
(such as Au, TiC or Ag nanocubes)? Actually we are looking for
nanoparticles with cubic shape morphology that are stable under the
electron beam (low knock-on damage). So far the best Nanoparticle we
could find is the Gold�Copper Bimetallic and Titanium carbide (TiC)
Nanocubes, but unfortunately we don't have the experience to
synthesize it!

Any Help You Can Give Will Be Greatly Appreciated!

Thank you

and wish you a good week start!
--
Ala' Afeef,
Glasgow University
*************************************************
Sir Alwyn Williams Building, Lilybank Gardens, University of Glasgow,
Glasgow, G12 8QQ, Scotland.

*************************************************

==============================Original Headers==============================
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7, 29 -- From: alaa afeef {alaa.afeef-at-gmail.com}
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From noreply-at-frois.com Sun Dec 1 18:57:22 2013
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Hello all,
Please, does anybody know the specification of rod seals used in
TEM MegaViewII camera (Olympus formerly Sis)?

Thanking you in advance.

Best regards
Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR. v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

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Dear everyone,

We have a very old PGT EDS system on our JEOL 2010 TEM. Recently we
would like to update it to a new one. After discussing with JEOL, they
say that Cooling fins are needed to help in reducing sample
contamination.

Since the Cooling fins are expensive, we just wondered whether the
cooling fins are really required for all EDS systems(Oxford, Bruker or
EDAX)? Does anyone know of any EDS system that does not require this
cooling-fin modification on the JEOL 2010 in order to be installed?

Thank you very much!

--
Dr. Yueling Qin
UB2020 Integrated Nanostructured Systems Initiative,
University at Buffalo, SUNY, Buffalo, NY 14260,
Phone (716) 645-8698

==============================Original Headers==============================
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Complete Novascan 30 SEM last in use in 2000 at Harbor Branch in Fort
Pierce, FL, believed to be in working condition

For a nominal fee will deliver to Boston area or crate for shipping longer
distances

Will consider sending parts if nobody wants the complete system

Please contact ekarplus-at-sciencewares.com for more information

Eric Karplus
508-457-4554 office
508-457-4556 cell
ekarplus-at-sciencewares.com

==============================Original Headers==============================
7, 29 -- From ekarplus-at-sciencewares.com Mon Dec 2 10:59:43 2013
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7, 29 -- Subject: Novascan 30 SEM system and/or parts
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Are there issues with postings done with firefox vs explorer?

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Hello Listserver,

We recently got some samples consisting of certain percentages of cellulose nanocrystals embedded in some sort of epoxy resin. I sectioned them at 70nm, and tried a couple of methods for "staining" the samples with OsO4. Vapor fixation and aqueous solutions were used for 1-2 hours, but I can't seem to get any definitive results from the samples. The contrast in the images is very poor, and I can't see anything that looks like what they are expecting to see. (Yes I've tried different objective apertures and the like on the imaging side of things.)

The most recent publication I could find on the subject was something from the 80's, and the methods section was not very clear on the TEM sample prep.

Does anyone out there have experience with imaging this type of sample?

Thanks,
Bradford Ross

Microscopy Technician
BioImaging Facility
University of British Columbia
6270 University Blvd.
Vancouver, B.C.
Canada
V6T 1Z4

phone 604-822-6996

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X-from: jmlebeau-at-ncsu.edu ()

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Email: jmlebeau-at-ncsu.edu
Name: James LeBeau

Organization: North Carolina State University

Title-Subject: [Filtered] Fwd: staff and postdoc positions at NC State's Analytical Instrumentation
Facility (AIF)

Message: See message below, sent on behalf of the Associate Director of the Analytical
Instrumentation Facility at NCSU.

------------------
Dear colleagues,

The Analytical Instrumentation Facility (AIF) at North Carolina State University is now seeking to
fill multiple staff and postdoctoral positions in the areas of electron microscopy and X-ray
diffraction. Positions are detailed in the links provided below. Please forward this information to
those who may have an interest in applying.

TEM facility manager: https://jobs.ncsu.edu/postings/30882
X-ray diffraction laboratory manager: https://jobs.ncsu.edu/postings/30892
Postdoctoral research scholars who are experts or emerging experts in at least one of the areas of
electron microscopy, X-ray diffraction, and/or surface analysis:https://jobs.ncsu.edu/postings/30668

Regards,
Jacob L. Jones

Associate Professor, Department of Materials Science and Engineering
Associate Director, Analytical Instrumentation Facility
North Carolina State University
Raleigh, NC
Email: JacobJones-at-ncsu.edu
Phone: 919-515-4557

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Brief answer to the recent request concerning nanocubes stable under
electron beam:

We can prepare Pd nanocubes (colloidal solution, average size 15nm,
stable both in the solution and in TEM at 120 kV).

TEM micrographs of the nanocubes and other details are in our paper
[Colloids and Surfaces B: Biointerfaces 100 (2012) 205� 208].

We prepare nanoparticles for other collaborating institutes. Feel free
to contact me if the nanocubes are suitable for you.

With best regards, Miroslav Slouf
---
Miroslav Slouf (slouf-AT-imc.cas.cz)
Institute of Macromolecular Chemistry AS CR, v.v.i.
Heyrovskeho namesti 2, 16206 Praha 6, Czech Republic

=== Original post ===

Hi Everyone!

Are you aware of a company or group that can provide some nanocubes
(such as Au, TiC or Ag nanocubes)? Actually we are looking for
nanoparticles with cubic shape morphology that are stable under the
electron beam (low knock-on damage). So far the best Nanoparticle we
could find is the Gold�Copper Bimetallic and Titanium carbide (TiC)
nanocubes, but unfortunately we don't have the experience to
synthesize it!

Any Help You Can Give Will Be Greatly Appreciated!
Thank you and wish you a good week start!
--
Ala' Afeef, Glasgow University

==============================Original Headers==============================
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I use Firefox (Mac 10.8.5) and have no issues except those caused by the
university IT people. Have not tried Explorer.

Phil

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} Are there issues with postings done with firefox vs explorer?
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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3, 29 -- Subject: Re: [Microscopy] firefox test
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Dear Michael,
I would like to comment on your query with my own experience for the last 60 mins:
the message below was posted via the

{Microscopy ListServer E-mail Submission Form} at www.microscopy.com

(using i) MOZILLA FireFox AND ii) afterwards Internet Explorer Vs.8.06... as indicated below):

========================================================================================================
i) local time: 14:46: (Nestor's time: 07:46:34, see below)
Email: w.muss-at-salk.at
Name: MUSS Wolfgang
Organization: SALK-LKH(Gen.Hospital SALZBURG, AUSTRIA)
Title-Subject: Re: firefox test [-at- delannoy-at-jhmi.edu]
Message: Just as an information to Michael Delannoy:
This message sent: via:
opening: MOZILLA-FireFox browser (Vs 25.0.1),

addressing for {www.microscopy.com} ,

opening: {Submit/Post a Message}
On-Line Form to
Submit a Question, Comment
to the Listserver

opened: {Microscopy ListServer E-mail Submission Form}

TEXT:
Sent for testing only Best regards and wishes Wolfgang MUSS SALZBURG / Austria (normally using Internet Explorer Vs 8.06,XP)
Then: ==} SUBMIT EMAIL...

After Online Submission immediate reply via:
http://www.microscopy.com/cgi-bin/NJZListServerEmailForm.pl?recipient=Zaluzec%40MICROSCOPY.COM&Subject=MicroscopyListserver-via-WWW%3A&required=Name%2CTitle-Subject%2CEmail%2CMessage&TITLE=Microscopy+Listserver+Form&sort=order%3AName%2CEmail%2COrganization%2CTitle-Subject%2CMessage&Name=MUSS+Wolfgang&Email=w.muss%40salk.at&Organization=SALK-LKH%28Gen.Hospital+SALZBURG%2C+AUSTRIA%29&Title-Subject=Re%3A+firefox+test+++[%40+delannoy%40jhmi.edu]&Message=Just+as+an+information+to+Michael+Delannoy%3A%0D%0A%0D%0AThis+message+sent%3A%0D%0A+%0D%0Avia%3A+%3Copening%3E+MOZILLA-FireFox+browser+%28Vs+25.0.1%29+%0D%0Acalling+for%0D%0A+%0D%0A%3Cwww.microscopy.com%3E%2C+%0D%0Aopening%3A+Microscopy+ListServer+E-mail+Submission+Form+%0D%0A%0D%0ATEXT%3A+Sent+for+testing+only+%0D%0A%0D%0ABest+regards+and+wishes%0D%0A%0D%0AWolfgang+MUSS%0D%0ASALZBURG+%2F+Austria%0D%0A%28normally+using+Internet+Explorer+Vs+8.06%2CXP%29%0D%0A%0D%0A%3D%3D%3E+SUBMIT+EMAIL...

Automatic Text display:

{ { { Thank You For Filling Out This Form
Below is what you submitted to Zaluzec-at-MICROSCOPY.COM on Tuesday, December 3, 2013 at 07:46:34

Email: w.muss-at-salk.at
Name: MUSS Wolfgang
Organization: etc. etc. } } }

------------

Note 1): This my message has not been received in my Outlook-box as a MSA-ListServer-Participant after 40 minutes waiting. It may be that everytime one uses the {Submit/Post a Message}
On-Line Form to
Submit a Question, Comment
to the Listserver,
the message will go through redaction/inspection of/by Nestor Zaluzec? If NOT, there might be a problem with using Mozilla Firefox, I guess....

Note 2): if a regular participant/registrated member on the MSA-ListServer, you always can post your messages/questions etc. (Cave: ONLY TEXT-FORMAT) via the e-mail-system you use, addressing the direct e-mail address
to: { microscopy-at-microscopy.com }

-----------------------------------------------------------------------------------
PS:
ii) The same done 15:26 local time = 40 minutes later than message 1 with Mozilla Firefox:

Just as an information to Michael Delannoy:
This message sent: via opening: Internet Explorer(Vs 8.06...XP),

addressing for {www.microscopy.com} ,

opening: {Submit/Post a Message}
On-Line Form to
Submit a Question, Comment
to the Listserver

opened: {Microscopy ListServer E-mail Submission Form}

inserted TEXT:
{ { { As above, +
Sent for testing only Best regards and wishes Wolfgang MUSS SALZBURG / Austria
(normally using Internet Explorer Vs 8.06,XP)
==} SUBMIT EMAIL... ==}

==} Automatic Reply from Online-Sever ('Zaluzec'):
{ { {Thank You For Filling Out This Form
Below is what you submitted to Zaluzec-at-MICROSCOPY.COM on Tuesday, December 3, 2013 at 08:27:16}

Email: w.muss-at-salk.at
Name: MUSS Wolfgang
Organization: SALK-LKH(Gen.Hospital SALZBURG, AUSTRIA)
Title-Subject: Re: firefox test [-at- delannoy-at-jhmi.edu]

Message: Just as an information to Michael Delannoy: This message sent: via: opening: Internet Explorer(Vs 8.06...XP), addressing for , opening: On-Line Form to Submit a Question, Comment to the Listserver opened: TEXT: Sent for testing only Best regards and wishes Wolfgang MUSS SALZBURG / Austria (normally using Internet Explorer Vs 8.06,XP) ==} SUBMIT EMAIL... } } }

So perhaps Prof. Nestor Zaluzec is not available still....

Best regards,
Wolfgang
(this mail sent 24 minutes post requesting via online submission form, no delivered e-mail-Message in my Outlook-box so far).

========================================================================================================

Von: oshel1pe-at-cmich.edu [mailto:oshel1pe-at-cmich.edu]
Gesendet: Dienstag, 03. Dezember 2013 14:20
An: Mu� Wolfgang
Betreff: [Microscopy] Re: firefox test: Are there issues with postings done with firefox vs explorer? // Re: firefox test

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I use Firefox (Mac 10.8.5) and have no issues except those caused by the
university IT people. Have not tried Explorer.

Phil

Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Are there issues with postings done with firefox vs explorer?
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

==============================Original Headers==============================
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44, 38 -- To: "'microscopy-at-microscopy.com'" {microscopy-at-microscopy.com}
44, 38 -- Subject: [Microscopy] Re: firefox test: // Re: firefox test
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44, 38 -- References: {201312031320.rB3DKIlb009037-at-ns.microscopy.com}
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Listservers,
Thanks for all your helpful suggesstions. If I can get this message out it
was HTML vs Plain text (thanks Nester).
We are switching our email provider.

Sincerely,
Micahel Delannoy

==============================Original Headers==============================
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Hello Listservers,

The question comes up again, what is the recommended image capture method
for cryo-tomography? Is it traditional EM film or digitized images?

For cryo-tomography work which microscope would be more suited for a
multi-user facility the FEI Tecnai Spirit (G2 ?) or the Hitachi H-7700? (no
vendors please).

thanks for your response,

Michael Delannoy

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Brad

I also tried to image cellulose in a hydrocarbon matrix with no luck. We finally asked the customer for a sol gel of the cellulose and were able to cut that on our Quanta 3D FIB and only then got some very nice tilt series and reconstructions. We got some help from FEI on the reconstructions as they are only 1 hour from us.

I am happy to share images with you and FIB tricks. It is very tricky to cut and lift em out in the FIB. But succeeded with a low KV ion beam and a PMMA recipe and judicious use of FIB protect layers.

Call or email and we can chat!

Pete Eschbach
Electron Microscope Facility Director
Oregon State University
Linus Pauling Science Center Suite 145
Corvallis, OR
97331

541 737 5645

Sent from my iPhone

} On Dec 2, 2013, at 2:50 PM, bradford.ross-at-botany.ubc.ca wrote:
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} We recently got some samples consisting of certain percentages of cellulose nanocrystals embedded in some sort of epoxy resin. I sectioned them at 70nm, and tried a couple of methods for "staining" the samples with OsO4. Vapor fixation and aqueous solutions were used for 1-2 hours, but I can't seem to get any definitive results from the samples. The contrast in the images is very poor, and I can't see anything that looks like what they are expecting to see. (Yes I've tried different objective apertures and the like on the imaging side of things.)
}
} The most recent publication I could find on the subject was something from the 80's, and the methods section was not very clear on the TEM sample prep.
}
} Does anyone out there have experience with imaging this type of sample?
}
} Thanks,
} Bradford Ross
}
} Microscopy Technician
} BioImaging Facility
} University of British Columbia
} 6270 University Blvd.
} Vancouver, B.C.
} Canada
} V6T 1Z4
}
} phone 604-822-6996
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Colleagues.....

All on-line form postings are delayed. The
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Your Friendly Neighborhood SysOp

Nestor

--
===========================================

Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
Bldg 212 / A-143
9700 S. Cass Ave
Argonne, Illinois 60439 USA

Tel: 530-NES-TORZ (530-637-8679) has Voice Mail
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Senior Scientist - Argonne National Laboratory
Fellow of the Microscopy Society of America
Senior Fellow the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
Past President Microscopy Society of America
Adjunct Professor of Physics - Northern Illinois University &
the University of Illinois at Chicago
Visiting Professor of Microscopy - Manchester University

===========================================
TPMLab: http://tpm.amc.anl.gov
MMSite: http://www.amc.anl.gov
===========================================

The box said ...
"This program requires Win 95/98/NT or better..."
So I bought a Mac !

===========================================

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Nestor,
Thank you for all you do, and especially for keeping the Spam Monster at
bay. I know my little tiny business email address gets more spam than
legitimate email daily (sometimes 2:1 or worse) and I truly admire your
dedication to keeping the Listserver spam free. The minor restrictions that
are in place are truly a joy compared to the alternative.

If I should ever have the pleasure of running into you, I will definitely
buy you a beer or three (not virtual ones).

Ken Converse
owner

QUALITY IMAGES
Servicing Scanning Electron Microscopes
Since 1981
474 So. Bridgton Rd.
Bridgton, ME 04009
207-647-4348
kenconverse-at-qualityimages.biz
qualityimages.biz

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Sent: Tuesday, December 03, 2013 3:27 PM
To: kenconverse-at-qualityimages.biz

Colleagues.....

All on-line form postings are delayed. The
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are from subscribers with validated Email addresses.

I personally read all on-line (i.e WWW form postings)
and approve them before forwarding them to the Listserver
and I generally only do this once/day over a cup or two
of coffee at Breakfast.

The reason for the delay is very simple. On-line forms are easily
exploited as spam generators. For example, this
morning before Wolfgang Muss used the form to post
his test messages there were 6 junk messages.

I'm pretty sure you were all not really interested
in reading the message from quirtz-at-gmail.com
about how nice his(her?) chocolates were and that
you could buy them for a reasonable price today only.
Or the offer of becoming a friend to a very pretty
person in a lonely suburb outside of Moscow....

The Listserver literally gets hundreds
of spam messages per day (this address has been on-line
for over 20 years now) and it unfortunately easy to
find and is on far too many lists that have been hijacked
by junk-mailers.

Fortunately (or not depending upon your point of view) I have managed to
create a reasonably effective system to mitigate and minimize
junk mail from getting through. It does cause delays,
and you all have to follow a fairly restrictive set of rules, but
that is the cost of doing business now adays.

Wolfgang, your message will likely get through the system
tomorrow AM when I review the daily log files.

Right... time to get back to my "other job". ;-)

Cheers

Your Friendly Neighborhood SysOp

Nestor

--
===========================================

Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
Bldg 212 / A-143
9700 S. Cass Ave
Argonne, Illinois 60439 USA

Tel: 530-NES-TORZ (530-637-8679) has Voice Mail
Lab: 630-252-7901
Fax: 630-252-4798

iChat: Zaluzec-at-AIM.com
Skype: Zaluzec-at-ANL
Polycom: 146.139.72.119
TPM: http://tpm.amc.anl.gov

Email: Zaluzec-at-aaem.amc.anl.gov

Senior Scientist - Argonne National Laboratory
Fellow of the Microscopy Society of America
Senior Fellow the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
Past President Microscopy Society of America
Adjunct Professor of Physics - Northern Illinois University &
the University of Illinois at Chicago
Visiting Professor of Microscopy - Manchester University

===========================================
TPMLab: http://tpm.amc.anl.gov
MMSite: http://www.amc.anl.gov
===========================================

The box said ...
"This program requires Win 95/98/NT or better..."
So I bought a Mac !

===========================================

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OK, this is sort of spam, but can only concur with Ken's sentiments.
Thanks much, Nestor!!
Definitely beers if/when you are back downunder.
best,
Rosemary White

Dr Rosemary White
CSIRO Plant Industry
GPO Box 1600
Canberra, ACT 2601
Australia

T 61 2 6246 5475
F 61 2 6246 5334
E rosemary.white-at-csiro.au

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I haven't been getting enough e-mail, so let me pose a question.

I'm having chiller troubles on my Phillips CM-12. The service man has decided to replace the thermostat, but that will take at least 7-10 days. Normally the column surface temperature at the water cooled lenses is 72F. Today they are running at 84F. I turned the scope off.

Question: Are there any studies or personal experience regarding changes in image magnification as a function of operating temperature and will I damage the scope operating at an elevated temperature? I know I'm more likely to crud up the column from the inadequate cooling at the diffusion pump, but what else could happen?

Thanks in advance!!

Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Position Opening at:

RESEARCH RESOURCES CENTER,
UNIVERSITY OF ILLINOIS AT CHICAGO

Visiting Research Specialist- Microscopy Specialist

The Electron Microscopy Service (EMS) of the Research Resources Center
(RRC) at the University of Illinois at Chicago has an open position for
a Visiting Research Specialist- Microscopy Specialist. The facility
provides electron and laser microscopy and surface analysis services for
the university research community and external organizations from two
sites on the campus. The open position is in the EMS-West facility,
which specializes in life science TEM and life and materials science
SEM. The west side facility includes a JEOL JEM-1220 TEM, Hitachi
S-3000N VPSEM and a JEOL JSM-6320F FESEM.

Qualified candidates must have a bachelors degree (preferably a
masters degree or higher) in a related field, with at least three years
electron microscopy experience in SEM. They will help supervise the
operation of the specimen preparation area, including record keeping and
maintenance and will assist/supervise in the day to day running of the
Electron Microscopes. Interpersonal/ Communications skills are
important, as this individual will work with users, provide technical
advice and demonstrate how microscopy can advance their research.

Further information may be obtained by consulting the following website:
http://www.rrc.uic.edu/

Applications must be submitted through the hire touch system at
https://jobs.uic.edu/default.cfm?page=job&jobID=33863 . For fullest
consideration, interested parties should send an application no later
than January 4, 2014 with a cover letter, complete curriculum vitae, and
the names and addresses of three references. Please address all
materials to Alan W Nicholls, Ph.D.

UIC is an EEO/AA.

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Frank:

84 Deg at the column is just one of the symptoms of inadequate cooling.
Aside from an overheated diffusion pump back-streaming hydrocarbons into the column and ion pump, the added overheating of water cooled electronics may result in untimely component failure and future repair down time.

If you overheat the lenses and short one out, the repair costs may exceed $10,000.
Think of it as a car....... Would you drive a 1/4 million dollar car that has a known and acknowledged cooling problem?

Check the temperature of your recirc. Water and if it is also hot, you can be assured that the entire microscope is being thermally stressed.

Most of the chiller manufactures stock critical parts and offer overnight/next day delivery.

Also, if you back-stream DP oil into the column, you will be haunted by poor gun vacuum, poor resolution and all the attendant problems associated with this evil.

Good luck!

Pierre Bustanoby
SEMS
Specialized
Electron
Microscope
Service

Serving the Pacific Northwest
Since 1975
206-795-1599

_________________________________

I haven't been getting enough e-mail, so let me pose a question.

I'm having chiller troubles on my Phillips CM-12. The service man has decided to replace the thermostat, but that will take at least 7-10 days. Normally the column surface temperature at the water cooled lenses is 72F. Today they are running at 84F. I turned the scope off.

Question: Are there any studies or personal experience regarding changes in image magnification as a function of operating temperature and will I damage the scope operating at an elevated temperature? I know I'm more likely to crud up the column from the inadequate cooling at the diffusion pump, but what else could happen?
Frank

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I thought I'd mention my modification to my TEM. Frankly I don't know why all TEM don't come this way.

I am always concerned about water temperature and the TEM Temp cause I have a crappy chiller prone to failure and excess.

So, I bought a bunch of liquid crystal thermometers, like you see stuck on the outside of fish tanks, from MC Master-Carr . They go from 58F(14C) to 88F (31C). (http://www.mcmaster.com/#temperature-indicating-labels/=pnwga7)

They come with a pressure sensitive adhesive, but I left the backing on so I could removed them and used clear packing tape to fix them on TEM column where the lenses are. I also taped one to the inlet and outlet on the water lines.

Now I can quickly find out what the surface temp is. It's not the actual temp, but for a $1.10 per gauge I know if I'm over heating or excessively cold.

Oh, I'm not connected with Mc Master-Carr. I don't have any friends who work there. They provide great service and if you tell them Frank sent you, they'll only charge you double. I have no pull with them.

Stay safe.............

Frank

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Title-Subject: [Filtered] Weight of Philips CM10

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Josh,

The values I find in the installation specs for weights in kilograms:

Console + column 990 kg
Power supply with HT generator 430 kg
HT generator 100 kg

Weight distribution 675 kg/m^2

From the "Space and Floor Loading requirements" page.

Let me know if you need pdfs of the installation specs - I can send you
those.

Phil

} Email: jcsmtf-at-mail.missouri.edu
} Name: Josh Schorp
}
} Organization: University of Missouri
}
} Title-Subject: [Filtered] Weight of Philips CM10
}
} Message: Does anyone have information on the weight of the CM10 and the high voltage unit? I can not
} find any information on net weights in the manuals.
--
Philip Oshel
Microscopy Facility Supervisor
Biology Department
024C Brooks Hall
Central Michigan University
Mt. Pleasant, MI 48859
(989) 774-3576

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Dear listers,
to Frank's problem on the CM12 water cooling system:
84 F (almost 30 C?) is too hot, for quite a few parts of the TEM. Yes, switching off and searching for the problem, and solving it, is recommended, IMO.
Many things can be wrong, on the thermostat, and here and there. Ours is not the first one any more, it was replaced once, in 23 years, and also the second one needed some maintenance (fair enough) and some repair, too.
In some cases, a careful check for microbes, yeasts, microbial mats, etc inside the water bath (and, consequently, inside all the tubings of the water circulating system) might be worth doing. It can happen that there is a really severe microbial contamination, building up over the years, resistant to all that "anti-microbial" stuff that you had thought should be in the water. Microbes can become very resistant to those substances which you think are poisonous - some even can live on it! in the dark, at or below 18 C, no light, no organics: no problem for them, if they have time. - Be aware, and have a check of an aliquot (possibly after a short centrifugation), with a high resolution phase contrast light microscope (if you have one). Phase contrast is recommended - then you do not need any fixation/staining. And you can see the tiniest bugs, happily swimming around. (if you do not see any: carefully check the light microscope - they can be overlooked!)
Yes, this happened to me -- not only once; and I am microbiologist ... ;-(
remedy? NO antibiotics, please!! But: many (5+ x) rinses with water, then 70% ethanol (for our CM12, this was ok, for about 30-60 min or so; do not forget to switch on the CM12, temporarily!), then 3 to 5 rinses with water, then again 70% ethanol, then many rinses with water again. The ethanol has to be removed - it is a nice food for some microbes (not only for us .........). Also, clean the internal part of the chiller rigorously. In the worst case, it might be necessary to clean some pipes (e.g. at the column) with pressurized air or ... replace them, in the worst case. Replace long tubings, if you can afford it, after some years. They might be an overseen reservoir of bugs in the form of biomats. - then check again for possible growth of microbes every month or every 3 months. Can be done in 5 minutes, only. And add the usual amount of this "anti-microbial" stuff, and pray that the bugs obey the rules.
Just a note, for the unlikely case that ...
Reinhard

--
Prof. Dr. Reinhard Rachel
University of Regensburg
Centre for EM / Anatomy
Faculty of Biology & Preclin. Med.
Universitaetsstrasse 31
D-93053 Regensburg - Germany
tel +49 941 943 2837, 1720
fax +49 941 943 2868
mail reinhard.rachel-at-biologie.uni-regensburg.de
office: VKL 3.1.29

Next microscopy conference:
http://www.imc2014.com/
18th IMC 2014 in Prague, Czech Rep.

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Dear Friends,

On a FEG(S)TEM, for example a Tecnai F20, what are the procedures to measure the STEM scanning probe size together with the probe current?

I measured the probe size at Nanoprobe mode before I pressed into STEM mode, because in STEM mode the CCD sees a diffraction pattern scanning not the static probe. I recorded some lines on CCD when using the beam shift wobbles the nano-probe. The FWHM was taken as the probe size at 400K magnification. I took the fluscreen read as the probe current. I end up with a ~1nm probe with over 3 nA current at condenser aperture 150 spot size1 89uA emission. Is that real?

Any comments to my procedures please?
Best regards,
Zhou
Dr Z Zhou
Loughborough University, UK

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Your numbers sound fair or close, based on my FEI FEG experience (CM200ST), although I obviously don't know if they are exactly right.

One thing that FEI does well and "some" vendors do poorly is that, while in STEM mode, you can release the diffraction button and see the direct probe on the florescent screen / TEM camera. Thus, you don't have to look at the nanoprobe spot size and then pop over to STEM, but instead, tune the probe in STEM mode, park the beam in the center of the field of view in TIA with the red circle/green crosshair tool, and release the diffraction button, which leaves the probe in STEM but changes the projectors to TEM. My major complaint with one particular non-FEI tool is that there's no way to see the sample plane instead of the diffraction plane during STEM. Ronchiograms are nice, but so is direct probe imaging.

Chad

---------------------
Chad M. Parish, Ph.D.
Research and Development Staff Member
Materials Science and Technology Division
Oak Ridge National Laboratory
Phone: 1 865 574 0092
Email: parishcm-at-ornl.gov

-----Original Message-----
X-from: Z.Zhou-at-lboro.ac.uk [mailto:Z.Zhou-at-lboro.ac.uk]
Sent: Friday, December 06, 2013 6:03 AM
To: Parish, Chad M.

Dear Friends,

On a FEG(S)TEM, for example a Tecnai F20, what are the procedures to measure the STEM scanning probe size together with the probe current?

I measured the probe size at Nanoprobe mode before I pressed into STEM mode, because in STEM mode the CCD sees a diffraction pattern scanning not the static probe. I recorded some lines on CCD when using the beam shift wobbles the nano-probe. The FWHM was taken as the probe size at 400K magnification. I took the fluscreen read as the probe current. I end up with a ~1nm probe with over 3 nA current at condenser aperture 150 spot size1 89uA emission. Is that real?

Any comments to my procedures please?
Best regards,
Zhou
Dr Z Zhou
Loughborough University, UK

==============================Original Headers==============================
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***************************************************************************************
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realname - Tracey Pepper
Email - tpepperisu-at-gmail.com
ORGANIZATION - Iowa State
EDUCATION - Graduate College
SUBJECT_OF_QUESTION - x-ray tomography
QUESTION - I have a grad student inquiry to my facility regarding the
use of "u-CT or x-ray microtomography, Nanotomograph, which can obtain
3-dimensional data of micro structure" which we do not have here. I am
looking for any facilities that have this and are a fee for service.

Thanks!
Tracey Pepper
Microscopy and NanoImaging Facility

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Quick, I need to know this in 20 minutes - Can I fix bacteria growing on
aluminum with osmium tetroxide? Or will I get some horrible reaction
and regret it? It seems to me I have avoided this in the past for some
reason, but I can't access that aprt of my brain right now...

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*
****************************************************************************

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Not 100% sure but I strongly suspect it is ok. Mercury salts do weird things with aluminum but I don't think osmium does.

Thomas E. Phillips, Ph.D
Professor of Biological Sciences
Director, Molecular Cytology Core
2 Tucker Hall
University of Missouri
Columbia, MO 65211-7400
573-882-4712 (office)
573-882-0123 (fax)
phillipst-at-missouri.edu

http://biology.missouri.edu/people/?person=88
http://www.biotech.missouri.edu/mcc/

-----Original Message-----
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Sent: Friday, December 06, 2013 12:53 PM
To: Phillips, Thomas E.

Quick, I need to know this in 20 minutes - Can I fix bacteria growing on aluminum with osmium tetroxide? Or will I get some horrible reaction and regret it? It seems to me I have avoided this in the past for some reason, but I can't access that aprt of my brain right now...

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*
****************************************************************************

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Also not 100% sure, but aluminium is easily oxidized and OsO4 a strong oxidizing agent!
I would expect the alu to be affected.
Jan

On 7/12/2013, at 8:05 am, PhillipsT-at-missouri.edu wrote:

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} Not 100% sure but I strongly suspect it is ok. Mercury salts do weird things with aluminum but I don't think osmium does.
}
}
} Thomas E. Phillips, Ph.D
} Professor of Biological Sciences
} Director, Molecular Cytology Core
} 2 Tucker Hall
} University of Missouri
} Columbia, MO 65211-7400
} 573-882-4712 (office)
} 573-882-0123 (fax)
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} Quick, I need to know this in 20 minutes - Can I fix bacteria growing on aluminum with osmium tetroxide? Or will I get some horrible reaction and regret it? It seems to me I have avoided this in the past for some reason, but I can't access that aprt of my brain right now...
}
} 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*
} ****************************************************************************
}
} ==============================Original Headers==============================
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Bradford:

Coming late to the party here. OsO4 does some staining of cellulose
in cell walls but the real staining comes from following up with lead citrate
staining. (The osmium serves as a mordent for the Lead).

Another suggestion I've come across is 2% OsO4 after treating with
sorbyl chloride (I have not tried this one).

I have used Barium Permanganate as a post section stain for cell
walls. Its messy but does a wonderful job at defining wall substructures. I do
not know the specific method of action but it may provide some contrast to
finding the cellulose particles.

It was originally published for Fungi but I have used it for both Fungi
and Plants.

Hoch, H. C. 1977. Use of permanganate of increase the electron opacity
of fungal walls. Mycologia 69:1209-2113.

On 2 Dec 2013 at 16:51, bradford.ross-at-botany.ubc.ca wrote:

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}
} We recently got some samples consisting of certain percentages of
} cellulose nanocrystals embedded in some sort of epoxy resin. I
} sectioned them at 70nm, and tried a couple of methods for "staining"
} the samples with OsO4. Vapor fixation and aqueous solutions were used
} for 1-2 hours, but I can't seem to get any definitive results from the
} samples. The contrast in the images is very poor, and I can't see
} anything that looks like what they are expecting to see. (Yes I've
} tried different objective apertures and the like on the imaging side
} of things.)
}
} The most recent publication I could find on the subject was something
} from the 80's, and the methods section was not very clear on the TEM
} sample prep.
}
} Does anyone out there have experience with imaging this type of
} sample?
}
} Thanks,
} Bradford Ross
}
} Microscopy Technician
} BioImaging Facility
} University of British Columbia
} 6270 University Blvd.
} Vancouver, B.C.
} Canada
} V6T 1Z4
}
} phone 604-822-6996
}
}
}
}
}
}
} ==============================Original
} Headers============================== 12, 31 -- From
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***************************************************************************************
Forwarded from "Ask a Microscopist"
Please remember that the person asking the question is likely
not a member the listserver, and
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as well as to the list.
Using the "reply" function in your email does *not* send your answer
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****************************************************************************************

realname - Steven Hunt
Email - shunt-at-unifrax.com
EDUCATION - Undergraduate College
SUBJECT_OF_QUESTION - software
QUESTION - We currently use 30 year old software and technology to
measure sub-micron diameter fiber. Can you recommend a
microscope/software package that will let us measure sub-micron diameter
fiber "on-the-fly", that is, take many measurements without capturing a
picture, just capturing data real-time?
Thank you for your time.

Steven

==============================Original Headers==============================
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Dear Microscopy Managers and Directors
Please share with your light and fluorescence microscopy core facilities.
Thanks to a grant from Olympus Corporation there are tuition
scholarships available graduate students and post-docs for the

Advanced Microscopy and Digital Imaging Workshop

January 17-20, 2012

University of California, Santa Barbara

Offered by the Neuroscience Research Institute (NRI) and Department of
Molecular, Cellular, and Developmental Biology (MCDB).

Registration closes soon, contact Mary Raven m_raven-at-lifesci.ucsb.edu
with scholarship information.
See for workshop information:
https://www.lifesci.ucsb.edu/mcdb/events/imaging_workshop/

--
Mary Raven
Microscopy Facility Director
Neuroscience Research Institute &
Molecular, Cellular & Developmental Biology
Biology 2 Building, room 5173
The University of California
Santa Barbara, CA 93106-5060

https://ucsb-microscopy.ucsd.edu/schedule/
http://www.lifesci.ucsb.edu/mcdb/research/facilities/microscopy/microscopy.html
Phone: (805) 893 8702
Fax: (805) 893 2005

==============================Original Headers==============================
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11, 20 -- Imaging Workshop in Jan 17-20 workshop in Santa Barbara, CA
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***************************************************************************************
Forwarded from "Ask a Microscopist"
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****************************************************************************************

realname - Georgianne Ciraolo
Email - Georgianne.Ciraolo-at-cchmc.org
ORGANIZATION - Cincinnati Children's Hospital Medical Center
EDUCATION - Graduate College
LOCATION - Cincinnati, Ohio
SUBJECT_OF_QUESTION - Elastin Staining of Plastic sections
QUESTION - I have an investigator who would like to stain for Elastin
on samples that are embedded in plastic. These sections would be used
for Bright Field Microscopy. Does anyone have any ideas or experience
with Elastin staining.

Thanks in Advance

Georgianne Ciraolo
Electron Microscopy Tech II
Department of Pathology

==============================Original Headers==============================
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Dear Listers,
I am in the market for a HAADF detector for a 300 kV scope. Has anyone compared the Fischione to the Gatan HAADF detectors. This will be in the camera block side port position.
Thanks,
Ken

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Hello!
http://buzoviprijatelji.hr/_34.studies.showed.html?opamuhjtab515820

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4, 43 -- To: zhengjing-at-ecust.edu.cn, takatsur-at-med.gunma-u.ac.jp,
4, 43 -- m.bozzali-at-hsantalucia.it, goodwill-at-wadsworth.org, r.parlato-at-dkfz.de,
4, 43 -- microscopy-at-microscopy.com, wittum-at-gcsc.uni-frankfurt.de,
4, 43 -- ahding-at-med.cornell.edu, rkuczenski-at-ucsd.edu, betty.tijms-at-gmail.com,
4, 43 -- a.orlacchio-at-hsantalucia.it, c.windisch-at-jacobs-university.de,
4, 43 -- ashively-at-lecom.edu, ksundin-at-capofficeproducts.com,
4, 43 -- knehaul-at-mail.usf.edu, lmccaule-at-nursing.upenn.edu,
4, 43 -- yasuhiro-at-mbs.med.kyoto-u.ac.jp, vemugant-at-neurosurg.wisc.edu,
4, 43 -- mariliseeb-at-yahoo.com.br, a.isaacs-at-prion.ucl.ac.uk,
4, 43 -- haiweixu2001-at-yahoo.com.cn, tone.tonjum-at-medisin.uio.no,
4, 43 -- lynn.carney-at-bms.com, aflowe-at-mail.usf.edu, leea-at-lundbeck.com
4, 43 -- Subject: FWD:
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[Microscopy] FWD: received just some minute ago from: { ronmervis-at-aol.com }

Dear all, just to inform you:
I have tried the only given URL:

http:/&/buzoviprijatelji.hr/_34.studies.showed.html?opamuhjtab515820

within the Listserver-message (by NOT Clicking but searching the
URL via Google Search.

Result: no documents could be found

Therefore this could be a SPAM or SCAM message...

Best regards
Wolfgang MUSS
EM-Lab,
Univ. Inst. Pathology
SAK-LKH&PMU
SALZBURG - AUSTRIA

==============================Original Headers==============================
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The problem of staining elastin bothered me a couple of months ago. I used to know how to do so when I was running a clinical E.M. lab about 5 years back, but then forgot the technique, because the last time I used it was more than a decade ago, and I needed to dust off some cobwebs. I also had photocopies of the technique, but left them behind when I changed facilities. After some snooping on the Internet, it came back to me, and I found 3 papers for you. I used tannic acid as a stain on my thin sections to visualize the elastin fibers. Back then (about 15 years ago) I didn't have the Internet resources to track down the original papers for the technique. Here are 3 good papers, one for pre-embedding staining, one for post-embedding staining, and one comparing both techniques:

Electron Dense Staining Affinities of Mouse Oxytalan and Elastic Fibers. T. A. Simmons and J. K. Avery, Journal of Oral Pathology 9:183-188, 1980 Pre-embedding method

Ultrastructural Visualization of Elastic Fibres with a Tannate-Metal Salt Method. Masato Kageyama, Minoru Takagi, Richard T. Parmley, Masaaki Toda, Hiroshi Hirayama, Yoshihisa Toda, The Histochemical Journal 17(1): 93-103, 1985. http://link.springer.com/article/10.1007%2FBF01003406 A post-embedding stain for thin sections

Two Techniques for Electron Opaque Staining of Elastic Fibres Using Tannic Acid in Fresh and Formalin Fixed Tissue. A. Haidar, T. A. Ryder, M. A. Mobberley, and J. S. Wigglesworth, Journal of Clinical Pathology 45(7): 633-635, 1992. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC495198/ Both a pre- and a post- embedding staining technique are compared in this paper.

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

________________________________________
X-from: oshel1pe-at-cmich.edu [oshel1pe-at-cmich.edu]
Sent: Thursday, December 12, 2013 2:35 PM
To: Haller, Edward

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realname - Georgianne Ciraolo
Email - Georgianne.Ciraolo-at-cchmc.org
ORGANIZATION - Cincinnati Children's Hospital Medical Center
EDUCATION - Graduate College
LOCATION - Cincinnati, Ohio
SUBJECT_OF_QUESTION - Elastin Staining of Plastic sections
QUESTION - I have an investigator who would like to stain for Elastin
on samples that are embedded in plastic. These sections would be used
for Bright Field Microscopy. Does anyone have any ideas or experience
with Elastin staining.

Thanks in Advance

Georgianne Ciraolo
Electron Microscopy Tech II
Department of Pathology

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

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Just to add to the valuable references given by Ed Haller, the following ones:

- An Electron Stain for Elastic Fibers using Orcein
Nakamura H, Kanai Ch, and Mizuhira V.
The Journal of Histochemistry and Cytochemistry,25 No 4, 306-308,1977 (Really a "Black pow(d)er! in elastic fibres)

- Rapid Contrasting of Extracellular Elements in Thin Sections
Koert P. Dingemans and Marius A. van den Bergh Weerman
Ultrastructural Pathology, 14:519-527, 1990
(FYI: my routine staining method since 1990/1991): 3-step staining method:
1. incubation of grids with sections down on drops of 0.01 - 0.05% hydrous Tannic acid (low mol. weight!) solution (micro-/Millipore filtered)
(RT to 60 degr.C.: e.g. original { Incubate in closed Petri dish in oven at 6O�C for three minutes} ),
followed by the usual double staining method ( UO2 ac. - Pb-citrate )
= original: {staining of sections with uranyl magnesium acetate and lead citrate in an LKB UltroStainer} :
yields great contrast improvement for collagen, elastin, as well as glycogen.

- A sulphur matrix complex, elastic fibril composed of a fine core of amorphous elastin and microfibrils was largely
accumulated in the aortic intima of aged rats.
Kazushige Ogawa* and Fumihiko Sasaki
Journal of Electron � Japanese Society of Microscopy Microscopy 52(2): 175-182 (2003)

(If you are in urgent need of the articles as pdf's, please don't hesitate to request them by e-mail)

Best wishes, good luck and
warm regards,

Wolfgang MUSS
SALZBURG, AUSTRIA

======================================================================================

Von: ehaller-at-health.usf.edu [mailto:ehaller-at-health.usf.edu]
Gesendet: Freitag, 13. Dezember 2013 15:01
An: Mu� Wolfgang
Betreff: [Microscopy] Re: Staining method for elastin embedded in plastic

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The problem of staining elastin bothered me a couple of months ago. I used to know how to do so when I was running a clinical E.M. lab about 5 years back, but then forgot the technique, because the last time I used it was more than a decade ago, and I needed to dust off some cobwebs. I also had photocopies of the technique, but left them behind when I changed facilities. After some snooping on the Internet, it came back to me, and I found 3 papers for you. I used tannic acid as a stain on my thin sections to visualize the elastin fibers. Back then (about 15 years ago) I didn't have the Internet resources to track down the original papers for the technique. Here are 3 good papers, one for pre-embedding staining, one for post-embedding staining, and one comparing both techniques:

Electron Dense Staining Affinities of Mouse Oxytalan and Elastic Fibers. T. A. Simmons and J. K. Avery, Journal of Oral Pathology 9:183-188, 1980
Pre-embedding method

Ultrastructural Visualization of Elastic Fibres with a Tannate-Metal Salt Method. Masato Kageyama, Minoru Takagi, Richard T. Parmley, Masaaki Toda, Hiroshi Hirayama, Yoshihisa Toda, The Histochemical Journal 17(1): 93-103, 1985. http://link.springer.com/article/10.1007%2FBF01003406
A post-embedding stain for thin sections

Two Techniques for Electron Opaque Staining of Elastic Fibres Using Tannic Acid in Fresh and Formalin Fixed Tissue. A. Haidar, T. A. Ryder, M. A. Mobberley, and J. S. Wigglesworth, Journal of Clinical Pathology 45(7): 633-635, 1992. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC495198/

Both a pre- and a post- embedding staining technique are compared in this paper.

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

________________________________________
X-from: oshel1pe-at-cmich.edu [oshel1pe-at-cmich.edu]
Sent: Thursday, December 12, 2013 2:35 PM
To: Haller, Edward

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realname - Georgianne Ciraolo
Email - Georgianne.Ciraolo-at-cchmc.org
ORGANIZATION - Cincinnati Children's Hospital Medical Center
EDUCATION - Graduate College
LOCATION - Cincinnati, Ohio
SUBJECT_OF_QUESTION - Elastin Staining of Plastic sections
QUESTION - I have an investigator who would like to stain for Elastin
on samples that are embedded in plastic. These sections would be used
for Bright Field Microscopy. Does anyone have any ideas or experience
with Elastin staining.

Thanks in Advance

Georgianne Ciraolo
Electron Microscopy Tech II
Department of Pathology

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

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Name: Robyn

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Title-Subject: [Filtered] Lead Citrate Staining

Message: I have been having a lot of problems with Lead citrate staining
and was wondering if anyone has found a really good recipe and technique
that doesn't have too many precipitate problems?

Currently I am using Lead nitrite and sodium citrate to create the lead
citrate stain...why? I dont know its just the protocol that was in place.

Thank you!!!

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Title-Subject: [Filtered] Job vacancy: Professional Officer Light and
Optical Microscopist at the University of Sydney

Message: Dear all,

The University of Sydney is currently seeking a Professional Officer
Light and Optical Microscopist. For more information and to apply, visit
sydney.edu.au/recruitment and search by the reference number 2440/1113.

PROFESSIONAL OFFICER LIGHT AND OPTICAL MICROSCOPIST
SYDNEY MICROSCOPY & MICROANALYSIS
DEPUTY VICE-CHANCELLOR RESEARCH
REFERENCE NO. 2440/1113

Expertise in light/optical microscopy essential
Full-time, continuing
Remuneration package: $92K p.a. which includes leave loading and up to
17% super

Sydney Microscopy & Microanalysis (SMM) aims to be a world-leading
facility for modern microscopy and microanalysis, offering premier
instruments, services and training to researchers from across Sydney and
around Australia, and providing strong leadership in the national and
international characterisation communities.

Reporting to the Laboratory Manager, you will provide high level support
to users across SMMs light and optical facilities, focussing primarily
on specialist techniques in microscopy, microanalysis, and analysis and
interpretation of images and data, to ensure these facilities provide
effective, high-quality outcomes for users.

In this role you will:

maintain the light/optical facilities in consultation with the Light
and Optical co-ordinator and extended team, the Laboratory Manager and
Directors
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training program for users groups in light/optical section
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develop a user portfolio
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related equipment and ensure implementation of WHS rules and regulations
develop and maintain good relations with relevant instrument and
equipment vendors.

To qualify for this role, you will demonstrate:

extensive training, experience and expertise in light-optical
microscopy and associated specimen preparation techniques and data analysis
skills in advanced light/optical techniques, including, FRET, FLIM,
FRAP, radiometric and live-cell, along with the associated analysis
techniques
experience in the servicing and maintenance of microscopes and
demonstrated ability to run and maintain sophisticated scientific
instrumentation
general knowledge with some other form of microscopy such as SEM, TEM,
AFM and their specimen preparation technologies
experience in the day-to-day running/operation issues of a multi-user
laboratory and experience in WHS and risk assessment preparation and
legislative requirements.

The successful candidate must possess a bachelors degree in science or
engineering. A postgraduate degree in this field will be highly regarded.

Remuneration package: $92,486 p.a. (which includes a base salary of
$78,152 p.a., leave loading and up to 17% employers contribution to
superannuation).

CLOSING DATE: 5pm 31 January 2014

The University is an equal opportunity employer committed to equity,
diversity and social inclusion. Applications from equity target groups
and women are encouraged. The University of Sydney has also established
a scheme to increase the number of Aboriginal and Torres Strait Islander
staff employed across the institution. Applications from people of
Aboriginal and Torres Strait Islander descent are encouraged.

© The University of Sydney

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Organization: NYU Abu Dhabi

Title-Subject: [Filtered] Embedding kit for electron microscopy

Message: Hello.I am purchasing supplies for our sample prep lab and
wanted to get some suggestions for an embedding material (kit?) for
general purpose electron microscopy. Suggestions?

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Hello Rita,
The yellow color is caused by reaction of glutaraldehyde with
buffer components (typically Tris or free amino acids; generally with
compounds possessing free amino groups). The best way is to avoid using
Tris based buffers for fixation.
If the buffer composition cannot be changed, you can use short
prefixation (~15 min, half strength of fixative) followed by short wash
with aldehyde compatible buffer and then use standard fixation
procedure in compatible buffer with full strength of fixative
(glutaraldehyde).

Please look at following books for details:
Principles and Techniques of Electron Microscopy: Biological
Applications. M. A. Hayat, Cambridge University Press, 2000 pp. 543

and/or

Fixation for Electron Microscopy (eBook Google)
M Hayat, Elsevier, 2. 12. 2012 - pp. 521

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

On Mon, 16 Dec 2013 17:59:20 -0600, zaluzec-at-microscopy.com wrote :
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} Title-Subject: [Filtered] Yellowed Glutaraldehyde
}
} Message: Hello fellow microscopists! Every now and then, we receive
} TEM samples from outside sources, in yellow glutaraldehyde. When we
} call to investigate why they are using yellow glutaraldehyde (and not
} requesting fresh fix), the answer is always "the glut was colorless
} when we collected and shipped the sample". Can anyone out there
} explain how or why the glut would change color in transport? Is it
} detrimental to the tissue? Will our results be compromised? Thanks
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Hi
I agee with Oldrich. There must be some amino acids or protein around but
thing is that it doesn't matter. It actually tells you that your client has
put some glutar.
Chers
yorgos

Dr Yorgos Nikas
Athens Innovative Microscopy
Skra 36 Voula 16673 GREECE

Tel/fax +30 210 8957677
mobile +30 6945 107477
www.eikonika.net www.aim.cat
*************************************
----- Original Message -----
X-from: {benada-at-biomed.cas.cz}
To: {eikonika-at-otenet.gr}
Sent: Tuesday, December 17, 2013 10:30 AM

Hello Robyn,
We are using "Lead Citrate Staining" as standard. The main problem with
precipitation is in CO2. Wet NaOH pellets should protect your sections
against CO2 from the air. However, you should also protect your
sections from direct breath. This is common mistake when students start
to play with "Lead Citrate Staining". It can be solved with some Safety
Face Shield.
Our technician is routinely wearing full face shield when doing "Lead
Citrate Staining".

I hope this can help you.

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

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} Title-Subject: [Filtered] Lead Citrate Staining
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} staining and was wondering if anyone has found a really good recipe
} and technique that doesn't have too many precipitate problems?
}
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Hello, Rita,

Having run a renal Pathology E.M. lab in prior years, I noticed that your e-mail address is listed as a clinic. By any chance, are you receiving clinical samples, and does your lab also receive samples in Michele's fix (Zeus transport medium)? If you receive samples in Michele's fix as well as samples in glutaraldehyde, the labs that are submitting samples to you in yellow glutaraldehyde are contaminating their glutaraldehyde with some Michele's fix when placing biopsies in their sample bottles. I was able to determine this years ago by careful tracking of the renal biopsies submitted to me from various hospitals. When I communicated the problem I was having to submitting renal Pathologists, and they stopped introducing a drop of Michele's fix into my glutaraldehyde bottles from forceps as they were obtaining biopsies, the problem was solved. The two solutions react to produce the yellowing and denaturing of the glutaraldehyde. In severely contaminated bottles of glutaraldehyde, the glutaraldehyde would actually turn milky yellow and would foam like soap if the bottle was shaken. The glutaraldehyde is cross-linking. Michele's fix is actually a transport medium without fixative properties, containing ammonium sulphate and n-ethylmaleimide. If cross-contamination of glutaraldehyde with Michele's fix is not the case, storing glutaraldehyde for prolonged periods of time under warm conditions can also cause the fixative to begin to polymerize and discolor, which is something else to investigate. Are submitting labs refrigerating their fixative, and using fresh fix?

Ed Haller

Edward Haller, Lab Manager
University of South Florida
Department of Integrative Biology
Electron Microscopy Core
SCA 110
4202 East Fowler Avenue
Tampa, FL 33620
(813)974-2676
ehaller-at-usf.edu
Office: ISA 1046
http://biology.usf.edu/ib/research/facilities/

________________________________________
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Email: kenner.rita-at-marshfieldclinic.org
Name: Rita Kenner

Organization: Marshfield Labs

Title-Subject: [Filtered] Yellowed Glutaraldehyde

Message: Hello fellow microscopists! Every now and then, we receive
TEM samples from outside sources, in yellow glutaraldehyde. When we
call to investigate why they are using yellow glutaraldehyde (and not
requesting fresh fix), the answer is always "the glut was colorless when
we collected and shipped the sample". Can anyone out there explain how
or why the glut would change color in transport? Is it detrimental to
the tissue? Will our results be compromised? Thanks in advance for your
help ... and Season's Greetings to all!

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Yes, Robyn, Oldrich is correct, and If I may chime in: In addition to holding your breath, boil all di-water ~15" (both stock reagent and di-rinse water), and use BOILING HOT di-water to DUNK clean BEFORE squirting grids clean with di-water. Should use 3 beakers of hot water (several dunks per beaker), not just one, and can even SKIP squirting them clean altogether. Boiling removes dissolved gasses (like CO2) and using it boiling hot rinses better, plus, tempers the sections a bit so the time required for the beam tempering procedure (tighten up the epoxy/reduce & eliminate drift) for the sections (some call it baking) can be reduced. I also believe that the surface tension in the dunking process helps too.

Work FAST, especially the part when the grid is transferred from the lead stain to the first hot water dunk. Hold your breath!

Technique based on Venable and Coggeshall, 1965.

Regards,
Andrea

Andrea Brothers
TEM Engineer 3
Yield Enhancement
Micron Technology
9600 Godwin Drive
Manassas, VA 20110
(703) 396-1836 direct
abrothers-at-micron.com

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Hello Robyn,
We are using "Lead Citrate Staining" as standard. The main problem with
precipitation is in CO2. Wet NaOH pellets should protect your sections
against CO2 from the air. However, you should also protect your
sections from direct breath. This is common mistake when students start
to play with "Lead Citrate Staining". It can be solved with some Safety
Face Shield.
Our technician is routinely wearing full face shield when doing "Lead
Citrate Staining".

I hope this can help you.

Best regards Oldrich

--
Oldrich Benada
Institute of Microbiology AS CR, v.v.i.
Videnska 1083
142 20 Prague 4
Czech Republic

On Mon, 16 Dec 2013 17:59:20 -0600, zaluzec-at-microscopy.com wrote :
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} Title-Subject: [Filtered] Lead Citrate Staining
}
} Message: I have been having a lot of problems with Lead citrate
} staining and was wondering if anyone has found a really good recipe
} and technique that doesn't have too many precipitate problems?
}
} Currently I am using Lead nitrite and sodium citrate to create the
} lead citrate stain...why? I dont know its just the protocol that was
} in place.
}
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} Thank you!!!
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The link worked for me, but it was an advertisement for a weight loss pill. Somehow, this got by Nestor's excellent filters.

John Mardinly, ASU

-----Original Message-----
X-from: W.Muss-at-salk.at [mailto:W.Muss-at-salk.at]
Sent: Friday, December 13, 2013 1:22 AM
To: John Mardinly

[Microscopy] FWD: received just some minute ago from: { ronmervis-at-aol.com }

Dear all, just to inform you:
I have tried the only given URL:

http:/&/buzoviprijatelji.hr/_34.studies.showed.html?opamuhjtab515820

within the Listserver-message (by NOT Clicking but searching the URL via Google Search.

Result: no documents could be found

Therefore this could be a SPAM or SCAM message...

Best regards
Wolfgang MUSS
EM-Lab,
Univ. Inst. Pathology
SAK-LKH&PMU
SALZBURG - AUSTRIA

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Colleagues....

Just in case your curious. The SPAM got through by a hacker
finding a user's account and password, logging into their account and
then sending blanket Email messages out to everyone in their
mailbox..

It is extremely difficult to design filters to stop that kind of attack, as it
comes from a validated user account. The user has contacted
me privately, verified the hijacking and has changed his password
so things should be returning to normal.

The obvious message is keep your Email accounts and logins secure.

Cheers
Nestor
Your Friendly Neighborhood SysOp

===========================================
Dr. Nestor J. Zaluzec
Argonne National Laboratory
Electron Microscopy Center
NanoScience and Technology/Bldg 212
9700 S. Cass Ave.
Argonne, Illinois 60439 USA
Tel: 530-NES-TORZ (530-637-8679) Fax: 630-252-4798

iChat:Zaluzec-at-AIM
Skype: Zaluzec-at-ANL
TPM: http://tpm.amc.anl.gov

Email: Zaluzec-at-aaem.amc.anl.gov
WWW: http://tpm.amc.anl.gov

Senior Scientist - Argonne National Laboratory
Fellow of the Microscopy Society of America
Senior Fellow of the Computational Institute - University of Chicago
E.P. Wigner Fellow - Oak Ridge National Laboratory
Past-President: Microscopy Society of America
Adjunct Professor - Northern Illinois University
Visiting Professor - Manchester University.
===========================================

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Hi everybody.

Is it possible that a very thin (membrane) protein crystal, consisting of maybe two or three layers of
proteins in there respective lipid membranes, could exhibit for example a p3 symmetry in zero degree
projection when the crystal as a 3-dimensional object doesn't have this symmetry?
In other words only the central slice of the Fourier transform parallel to the crystal plane would have 3-fold symmetry, but not a non-central slice.

Thanks and Merry Christmas,

Philip

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For the last two years I have been running laps on a mobius strip over TEM traceable calibration. Yes, I know about Mag-i-cal, but it's too expensive and fragile for us.

Well, I'm off the loop. We had our AL2A inspection. We passed the TEM carbon black determination portion.

Part of the problem was as a chemist and Microscopist I define calibration differently. I'll try to explain AL2A position.

Putting the grating replica in the scope and adjusting the imaging system to make acurate measurements is an adjustment. As far as A2LA is concerned that's a black box operation and what you do and how you do it is of no concern.

The calibration comes when you check the response (measurement) to a signal. To do this you put a known in the scope and measure it and ask yourself the question, "Is the result meaningful to my purpose?"

I know, it sounds confusing. As I see it, if you need to measurement object to 5 nm and your response is 1.5nm you're good to go. But is your response is 10nm, when how could measure something to 5nm?

So what's the response I'm talking about? How about the average of NIST traceable nanospheres?

What we have been doing is monthly adjusting the TEM imaging system with the grating replica and then checking the adjustment by making 100-200 measurements of 203nm Nanospheres. After a few months we had enough data to determine our uncertainty at k=2 and 95% confidence levels. The calculations are easy, like tripping down the stairs.

Poof!
In one fell swoop we are using a traceable calibration and determining our uncertainty measurement. We meet the A2LA criteria for using a traceable calibrated standard

If our response or average nanosphere measurement is too far off what do we do? We just (I'm told) readjust with the grating replica and redo our nanosphere measurements.

Long story a lot shorter. If you need a NIST-like traceable standard to calibrate your TEM, use traceable nanospherers to calibrate your TEM adjustment.

I don't work for the people who make Nanosphere, just so you know.

This email and any of its attachments may contain confidential information intended only for the use of the addressee(s). If the reader of this email is not the intended recipient or the employee or agent responsible for delivering it to the intended recipient, you are hereby notified that any dissemination or copying of this email is strictly prohibited. If you have received this email in error, please notify us by return email at info-at-ardl.com, permanently delete the email, and destroy any printouts. If this email contains test data and/or draft reports, you are hereby notified that only a signed original test report will contain official results, a copy of which resides in the project folder located at ARDL, Inc. Thank you. Akron Rubber Development Laboratory, Inc.

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Yes, maybe. I was thinking in that direction.

I get the impression that the software for 2D crystallography (mrc, 2dx, crisp etc.)
doesn't seem to allow for such a case. Is that true?

Philip

-----Original Messages-----

HI Philip,

could you have a screw axis, which in projection looks like it's 3-fold?

Mike
________________________

Hej Philip,

En tunn p31-kristall?

Merry Xmas,

Martin Lindahl

On Dec 20, 2013, at 8:44 AM, Philip K�ck {Philip.Koeck-at-ki.se} wrote:

} Hi everybody.
}
} Is it possible that a very thin (membrane) protein crystal, consisting
} of maybe two or three layers of proteins in there respective lipid
} membranes, could exhibit for example a p3 symmetry in zero degree projection when the crystal as a 3-dimensional object doesn't have this symmetry?
} In other words only the central slice of the Fourier transform parallel to the crystal plane would have 3-fold symmetry, but not a non-central slice.
}
} Thanks and Merry Christmas,
}
} Philip
} _______________________________________________
} 3dem mailing list
} 3dem-at-ncmir.ucsd.edu
} https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem

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Title-Subject: [Filtered] CM10 wire

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Hello Hitachi experts,

Main screen of H-7500 TEM will not lift/move when TEM is in diffraction
mode.

TEM in imaging mode - screen lifts up and image can be acquired by CCD
camera.

TEM in diffraction mode: I press screen lift button and hear a beep, but
screen will not lift up.

TEM in imaging mode - I lift the screen and then try switching to
diffraction mode. Hear a beep instead and TEM will remain in imaging mode.

In other words I could not achieve 2 conditions at the same time:
diffraction mode and screen "up". Problem is not relevant to beam
intensity or film camera settings or screen controls that are
accessible via standard operator menu, manual exposure setting instead
of automatic, etc. Operator manual has no mention of this condition.

Any ideas?

--
Vitaly Feingold
SIA
2773 Heath Lane
Duluth GA 30096
Ph. 770-232-7785
Fax 770-232-1791
www.sia-cam.com
vitaly-at-sia-cam.com

==============================Original Headers==============================
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Hi Vitaly,

Sounds like a "safety" interlock issue to keep from damaging the
camera/scintillator with the direct beam while in diffraction mode.
(caveat: I have no direct experience with the Hitachi scope)

Henk

At 12/21/2013 1:00 PM, vitalylazar-at-att.net wrote:
}
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}
} Hello Hitachi experts,
}
} Main screen of H-7500 TEM will not lift/move when TEM is in diffraction
} mode.
}
} TEM in imaging mode - screen lifts up and image can be acquired by CCD
} camera.
}
} TEM in diffraction mode: I press screen lift button and hear a beep, but
} screen will not lift up.
}
} TEM in imaging mode - I lift the screen and then try switching to
} diffraction mode. Hear a beep instead and TEM will remain in imaging mode.
}
} In other words I could not achieve 2 conditions at the same time:
} diffraction mode and screen "up". Problem is not relevant to beam
} intensity or film camera settings or screen controls that are
} accessible via standard operator menu, manual exposure setting instead
} of automatic, etc. Operator manual has no mention of this condition.
}
} Any ideas?
}

--
Untitled Document

Hendrik O. Colijn
*C*enter for *E*lectron *M*icroscopy & *A*nalysi*S*
1305 Kinnear Rd, Suite 100, Columbus, OH 43212

614.643.3458 Office colijn.1-at-osu.edu {mailto:colijn.1-at-osu.edu}
www.cemas.osu.edu {http://www.cemas.osu.edu}

"Time is that quality of nature which keeps things from happening all at
once. Lately it doesn't seem to be working."

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Email: Parmiterd-at-mail.nih.gov
Name: David

Organization: Leidos

Title-Subject: [Filtered] Non-Specific Antibody Interaction

Message: Hello all!

I am having a bit of a problem, and would like to know if anyone has any
suggestions as to how I could possibly improve my results:

I am attempting to label two antigens in the same sample.
Unfortunately, both these antigens are monoclonal mouse, though one is
IgG1 and the other is an IgG2.

Both appear to bind fine when I put them into a sample with both
antigens present, though there is no appreciable change to the label
distribution when the same process is applied to a sample that
supposedly has only one of the antigens present.

If anyone can supply some thoughts as to how I might increase binding
specificity in a sample, that'd be great. We've tried using a high salt
washing buffer for this, as well as trying to pre-bind the primary Abs
to their markers, though nothing seems to change our results.

Any suggestions are greatly appreciated! Thanks!

- David

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On Dec 21, 2013, at 10:11 AM, vitalylazar-at-att.net wrote:

} Hello Hitachi experts,
}
} Main screen of H-7500 TEM will not lift/move when TEM is in
} diffraction
} mode.
}
} TEM in imaging mode - screen lifts up and image can be acquired by
} CCD
} camera.
}
} TEM in diffraction mode: I press screen lift button and hear a beep,
} but
} screen will not lift up.
}
} TEM in imaging mode - I lift the screen and then try switching to
} diffraction mode. Hear a beep instead and TEM will remain in imaging
} mode.
}
} In other words I could not achieve 2 conditions at the same time:
} diffraction mode and screen "up". Problem is not relevant to beam
} intensity or film camera settings or screen controls that are
} accessible via standard operator menu, manual exposure setting instead
} of automatic, etc. Operator manual has no mention of this condition.
}
} Any ideas?
}
} --
} Vitaly Feingold

Dear Vitaly,
Disclaimer: I am not a Hitachi expert. There may be an interlock to
prevent the CCD from being damaged by the unscattered central beam in
diffraction mode. Some instruments have a more sophisticated system
which allows an ED pattern to be acquired on the CCD when the
intensity is sufficiently reduced, and some warn the user to do so.
You could try reducing the intensity to the point that the central
spot is no brighter than a bright area in imaging mode, set the
exposure time long enough to record scattered intensities of interest,
then try to remove the screen. Maybe the actual Hitachi experts will
have better ideas.
Yours,
Bill

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Hello all,

**(David informed me off list as follows:

Our primary Ab's are mouse-derived monoclonal AB's IgG1 and 2a. The secondary Ab's we're using are Goat-anti-Mouse IgG1 and Rabbit-anti-mouse IgG2a, and we label these with (gold conjugated) Tertiary Ab's, Donkey)**

A three step approach using donkey conjugates sounds ok, but it all depends of course on how specific the individual components are. Have you tested this? I am thinking of a simple dot-spot test, but you could also do this on specimens.
To solve the issue I would start from the tail of the incubation procedure (the gold conjugates) and work forwards towards the primaries…..

so.. the procedures would be as follows. All controls should of course be negative.

1. controls where the specimens are incubated with only the gold conjugates, separately as well as together.
if those are ok, then
2. controls where the specimens are incubated
a. with secondary1 and gold1 (matching pair)
b. with secondary2 and gold2 (matching pair)
c. for cross reactivity between secondary and gold conjugates:
secondary1 and gold2
secondary2 and gold1
d. for any reactivity between primary and gold conjugates:
primary1 and gold1
primary1 and gold2
primary2 and gold1
primary2 and gold2
if these are ok, then
3. cross reactivity tests: primary1, secondary2, gold2
primary2, secondary1, gold1

It is tedious, but with three step incubations and double labeling it can not be avoided if one wants reliable answers. This will be the way to establish specificity on specimens. The dot spot tests would be based on the same approach.

Hope this helps,

Merry Christmas and a happy new year to all!

Jan Leunissen
Aurion
i: www.aurion.nl

On 24/12/2013, at 11:19 am, Parmiter, David (NIH/NCI) [C] {parmiterd-at-mail.nih.gov} wrote:

} Hello, Jan -
}
} Thanks for your response. We're actually not working with tissue, but some manufactured material that expresses two viral proteins. Our primary Ab's are mouse-derived monoclonal AB's IgG1 and 2a. The secondary Ab's we're using are Goat-anti-Mouse IgG1 and Rabbit-anti-mouse IgG2a, and we label these with Tertiary Ab's, Donkey-anti-whatever. From Aurion, actually.
}
} Any suggestions you can offer would be greatly appreciated.
}
} Thanks kindly!
}
} David Parmiter [Contractor]
} Research Associate II
} Leidos Biomedical Research, Inc.
} Formerly SAIC-Frederick
} Frederick National Laboratory for Cancer Research
} P.O. Box B, Frederick, MD 21702
} Phone: 301-846-5990
} parmiterd-at-mail.nih.gov
}
}
}  Be Environmentally Responsible. Please don't print this e-mail unless you really need to
}
} This e-mail and any attachments to it are intended only for the identified recipients. It may contain proprietary or otherwise legally protected information for Leidos Biomedical Research, Inc. Any unauthorized use or disclosure of this communication is strictly prohibited. If you have received this communication in error, please notify the sender and delete or otherwise destroy the e-mail and all attachments immediately.
}
} -----Original Message-----
} From: Jan Leunissen [mailto:leunissen-at-aurion.nl]
} Sent: Monday, December 23, 2013 4:45 PM
} To: Parmiter, David (NIH/NCI) [C]
} Subject: non-specific antibody staining
}
} Hello David,
}
}
} I am replying to the message Nestor Zaluzec left on your behalf on the MSA list server.
}
} A few question for now to start off with:
}
} What is the tissue you work on? What species?
} What are the primaries directed against?
} What are the controls like? I.e. just secondary-Au. I assume you use a Goat-anti-Mouse IgG?
}
}
} Jan
}
} Aurion

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Hi,
In general a 2D crystal with any higher order symmetry such as P3 symmetry will give you a (P3) symmetric projection map when imaged without tilt, but the projection map of such a crystal from a slightly tilted direction will not have any symmetry. This already applies for single-layered 2D crystals, and also applies to multi-layered 2D crystals or 3D crystals. For true 3D crystals, there might be other (e.g., orthogonal) views where other symmetries can be found in projection. But this doesn't apply to 2D crystals.
If a structure has a screw axis, such as P312 or P321, then the structure is also P3 symmetric, and the same as above applies.

Philip, I think you are right: If you have a non-P3 symmetry crystal, but place three layers of such a crystal on top of each other, rotated each by 120 degrees with respect to each other, but centered onto the same point of origin, then the non-tilted projection map can well show P3 symmetry, even though none of the layers itself had that symmetry. In this case, processing that object under P3 symmetry would be wrong.

Besides this, there are possible other sources of a fake P3 symmetry appearance:
Hexagonally closest packing is a common way for proteins to squeeze together in a membrane. At low resolution, one sometimes gets the impression to have P3 or even P6 symmetry in such a case. But the low resolution might not allow conclusions anyway.
Another way to get apparent symmetries is when the crystal is fragmented, so that different ares in the 2D crystal have differently oriented lattice vectors. If, for example, you are dealing with a P2 symmetry crystal that has as lattice vectors a=100A, b=100A, and an included angle of gamma=120deg, and this crystal has cracks after which the lattice orientation changes so that the new b becomes the old a vector, then the overall appearance of such a crystal can be P3, even though the local areas aren't P3 symmetric.

All the best,

Henning.

Henning Stahlberg, PhD
Prof. for Structural Biology, C-CINA, Biozentrum, University Basel
Mattenstrasse 26 | D-BSSE | WRO-1058 | CH-4058 Basel | Switzerland
http://c-cina.org | Tel. +41-61-387 32 62

On Dec 20, 2013, at 3:26 PM, Philip K�ck wrote:

} Yes, maybe. I was thinking in that direction.
}
} I get the impression that the software for 2D crystallography (mrc, 2dx, crisp etc.)
} doesn't seem to allow for such a case. Is that true?
}
} Philip
}
}
} -----Original Messages-----
}
} HI Philip,
}
} could you have a screw axis, which in projection looks like it's 3-fold?
}
} Mike
} ________________________
}
} Hej Philip,
}
} En tunn p31-kristall?
}
} Merry Xmas,
}
} Martin Lindahl
}
}
} On Dec 20, 2013, at 8:44 AM, Philip K�ck {Philip.Koeck-at-ki.se} wrote:
}
} } Hi everybody.
} }
} } Is it possible that a very thin (membrane) protein crystal, consisting
} } of maybe two or three layers of proteins in there respective lipid
} } membranes, could exhibit for example a p3 symmetry in zero degree projection when the crystal as a 3-dimensional object doesn't have this symmetry?
} } In other words only the central slice of the Fourier transform parallel to the crystal plane would have 3-fold symmetry, but not a non-central slice.
} }
} } Thanks and Merry Christmas,
} }
} } Philip
} } _______________________________________________
} } 3dem mailing list
} } 3dem-at-ncmir.ucsd.edu
} } https://mail.ncmir.ucsd.edu/mailman/listinfo/3dem
}
} _______________________________________________
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} 3dem-at-ncmir.ucsd.edu
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From noreply-at-cinemadeamo.com Wed Dec 25 00:16:54 2013
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Hi, David:

You can also try the procedure below (Blocking and wash steps are not
included):

1. Incubate in primary antibody 1
2. Incubate in Ultrasmall gold particle conjugated F(ab) or F(ab')2
fragment of goat anti-mouse
(H&L).
3. Silver enhancement.
4. Incubate in primary antibody 2
5. Incubate in 6 or 10 nm gold particle conjugated goat anti-mouse.

The idea is when silver enhancing the first gold conjugate, the
antibodies
would be "encapsulated" by the silver nucleated around the Ultrasmall gold
particles
therefore would not be available for the second secondary (goat anti-mouse
as well) to bind.

I have never tried this procedure. If you decide to try, you should do a
control by incubating in 6 or 10 nm gold particle conjugated goat
anti-mouse
antibody after silver enhancement (without second primary). If you do not
see 6 or 10 nm gold particles on the sections, then that means the first
primary antibody is completely "encapsulated".

I am curious about the result if you try this procedure.

Happy New Year.

Hong

On 12/23/13 7:01 PM, "leunissen-at-aurion.nl" {leunissen-at-aurion.nl} wrote:

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