Three Colour Compensation

Alice.L.Givan@Dartmouth.EDU
29 Sep 92 13:13:25 EDT

Thanks to Marty Bigos for his comfirmation that I haven't been making up a
compensation problem that doesn't actually exist.

I like the suggestion about changing the FL3 filter to a longer wavelength so
as to lower the required compensation between PE and FL3. A 685 filter for
FL3 wouldn't really be a problem even using PI for DNA analysis -- as the FL2
filter (585nm) on the Scan is even better for PI than is the existing FL3
filter (650 LP), so most DNA analysis takes place through the FL2 channel.

While I am writing, does anyone have much experience with PI (for live/dead
gating) in conjunction with PE. I have a research group that really worries
about losing PE sensitivity for detecting dull cells if PI is present. My
own feeling is that, assuming you compensate correctly and assuming you, in
any case, gate OUT any cells that are PI-positive, then the PI should have
no effect on detection of weak PE signals. Any thoughts on this?
Thanks.
Alice
Alice Givan@Dartmouth.Edu

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