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Date: | Wed, 5 May 2004 13:05:03 -0400 |
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Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Folks I sent this a few days ago, but it bounced because of an email address
confusion, hopefully the info is still current and useful.
We run our Ultraview system through Metamorph, it works perfectly, however
having said that you will be able to use almost any of the high end scope
packages to do the same job. As the components within the system are fairly
generic/popular including Sutter filter wheels, PIfoc Z-drives, and
Hamamatsu cameras (which are core components on our system at least) the
marriage is seamless. The use of a high end scope management package is
really important if you need to drive components beyond those controlled by
the PE software, for example an XY stage or perhaps an automated scope.
Essentially you need to drive two filter wheels, one on the excitation side
which sorts the laser light and one on the emission side. The only thing
you need to do is to confirm with your PE rep where the filters are located
within the scanhead, on our system the excitation is position 0=blank,
position 1=488/10, 2=568/10 3=blank, 4=647/10. don't forget this will be
different if you are not using an argon/krypton source. Note there is no
physical shutter, essentially you choose position 3 on the filter wheel as a
shutter (this is done in the setup for meta). On the emission side 1=500nm
long pass (not used much) 2=525/50 (for GFP) 3=600/45 (rhodamine etc)
4=blank, 5=700/75, again this will change if you are using one of the more
complex 5 line systems. when you drive the filters in meta you can run them
at the fastest (0) speed, primarily because the wheels are relatively
lightly loaded. You can drive the aotf in the nifty new fast system but I am
not sure of the exact details. Laser power cannot be controlled because
there is no ND wheel or AOTF except on the newest systems. Generally this
is not a problem, you just decrease the exposure time in really bright
specimens.
simon
-----------------------------------------------------------
Simon C. Watkins Ph.D. FRC Path.
Professor, Cell Biology and Physiology
Director Center for Biologic Imaging
BSTS 225
University of Pittsburgh
3500 Terrace St.
Pittsburgh PA 15261
Tel:412-648-3051
Fax:412-648-2797
URL: http://www.cbi.pitt.edu
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