From: McCloskey, Thomas (thomasm@nshs.edu)
Date: Fri Sep 12 2003 - 15:47:29 EST
I am having a problem reading APC and PerCP together when using a 4 colour
system for identifying DC subtypes in peripheral blood. Although it is
clear that I have stained successfully in both HLA-DR:PerCP and CD11c:APC, I
am having dificulty getting a filter combination that will adequately read
both APC and PerCP together.
The cytometer is: Coulter epics elite
Hi Bernadette,
It is difficult to read percp adn apc together on an Elite due to
limited compensation circuitry and cross beam excitation. You have to
measure both these fluorochromes on the same PMT using aux channels and then
can't compensate for the PE signal.
I would recommend trying to use Fitc/PE/ECD/APC as your 4 color
combination on the Elite. ECD is available from Coulter adn flouresces at
about 610nms. If you separate the laser beams in time using gated
amplification, then this combination will work.
Good luck,
Tom
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Thomas W. Mc Closkey, Ph. D.
Director of Flow Cytometry, North Shore University Hospital
Assistant Professor of Pediatrics, New York University School of Medicine
Boas Marks Biomedical Research Center, 350 Community Drive
Manhasset, Long Island, New York 11030
ph: 516-562-4844 [office], 516-562-1135/4641 [lab] fax: 516-562-2866
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