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1k x 1k is too noisy for live cell on spinning disk. 512 is very lovely,
good s/n, but so tiny!!
Clare M. Waterman-Storer, Ph.D.
Associate Professor
Laboratory for Cell Motility Studies
Department of Cell Biology CB163
The Scripps Research Institute
10550 North Torrey Pines Road
La Jolla CA, 92037 USA
Office: 858-784-9764
Lab: 858-784-9243
Fax: 858-784-9779
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Administartive Assistant: Denise Munoz
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-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Jan Peychl
Sent: Tuesday, January 24, 2006 5:23 AM
To: [log in to unmask]
Subject: Re: EMCCDs for spinning disc ?
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Dear all,
following the question from Kate and answer from Clare, I would like to
ask you if you could share your experience with EMCCDs used with
spinning disc setup.
Do you already use these cameras for your spinning disc systems ? These
cameras are supposed to be fast which might be an advantage for spinning
disc applications.
But I am wondering what is your practical experience with speed, S/N
ratios and resolution of these cameras.
Did you have a chance to compare 512x512 chip versus 1kx1k chip when
it comes to the live cell imaging of biological samples
like mammalian cells, Drosophila imaginal disc or C. elegans embryos ?.
Any feedback would be very welcome , since we have so far very limited
experience
with these cameras. We also have a feeling ( no proper measurements)
that 1k*1k chip is noisier than 512*512 chip and when it comes to the
resolution,
then we have same a similar experience as Clare.
Thanks
Jan
xxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxxx
Jan Peychl, M.D., Ph.D.
Service Leader
Light Microscopy Facility
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108
01307 Dresden
Germany
Tel.: +49 351 210 2502
Fax: +49 351 210 2000
web: www.mpi-cbg.de
Clare Waterman wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Kate -We use the same camera as you for through the objective TIRF system.
>I love it, low noise, tiny pixles, and a lot of them. We also recently got
>a roper 512b for the same tirf system. it is great for tirf because the
>signal to background in the image is so great to begin with- not as good on
>spinning disk where contrast is not as good as tirf. The exposure times on
>orca II ERG for a speckle image were ~500 ms, with the 512b they are on the
>order of 25 ms to get the same speckle modulation. The main problem issue
is
>that the pixels are so big on 512b compared to orca type (~13 vs ~6 um)
that
>we had to modify the microscope to get nyquist satisfied. With all that
mag
>and only a 512 chip, we end up seeing only a very very small area of a
cell.
>I have been bugging camera companies for more real estate on those EM
chips.
>The 1k x1k em chip is too noisy compared to the 512.
>
>Don't know if any of this helps
>
>Clare M. Waterman-Storer, Ph.D.
>Associate Professor
>Laboratory for Cell Motility Studies
>Department of Cell Biology CB163
>The Scripps Research Institute
>10550 North Torrey Pines Road
>La Jolla CA, 92037 USA
>Office: 858-784-9764
>Lab: 858-784-9243
>Fax: 858-784-9779
>[log in to unmask]
>Administartive Assistant: Denise Munoz
>[log in to unmask]
>858-784-9964
>
>-----Original Message-----
>From: Confocal Microscopy List [mailto:[log in to unmask]] On
>Behalf Of Kate Luby-Phelps
>Sent: Monday, January 23, 2006 3:36 PM
>To: [log in to unmask]
>Subject: EMCCDs
>
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>What's the current wisdom on EMCCDs for TIRF microscopy? We are currently
>using a Hamamatsu
>Orca II ERG which performs remarkably well. However, we would like to
>improve the sensitivity of
>detection without sacrificing too much spatial resolution or S/N.
>
>Thanks,
>
>Kate L-P
>
>
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