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The use of PerCP does not necessarily change the need for FL1-FL3
compensation. The need for this compensation arises from the spectrum
of FITC, not CyChrome--no need to switch from CyChrome to the less
bright PerCP!
The fact is, "we" have been ignoring the need to do FL1 to FL3
compensation all these years, because we are lucky--by properly
compensating PE into FL3, and FITC into FL2, you are usually taking
care of the FITC into FL3 compensation need. This is explained in
detail in one of my Web pages on compensation: see the page
http://cmgm.stanford.edu/~roederer/compensation/3colorComp.html
If you have problems with FITC spilling into the third channel, your
only real option is to try different filters on the third channel until
the problem goes away. The redder a filter you can use, the better; we
use a 682/33 for the Cy5PE emission, together with a 585/15 for PE and
a 530/30 for FITC; this combination does not need significant
FITC-Cy5PE compensation.
> BD has promised to throw in calibrite beads capable of doing >
the three color compensation. Comments?
Well... BD wants to sell you PerCP reagents.
mr
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