Re: 7-AAD,FITC and PE

From: Joseph Trotter (trotter@scripps.edu)
Date: Tue Jan 27 1998 - 19:16:57 EST


Joan,

	You must compensate FL1 (FITC) signals out of FL3 (7-AAD) on most
cytometers indirectly. For example: On BDIS instruments compensation is
via dual differential amplifiers where you sample FL1 and remove a percentage
from FL2, sample FL2 and remove a percentage from FL1, and so on among 
adjacent channels. If you first balance FL1, FL2, and FL3 in terms of setting a 
similar sensitivity, then run compensation controls for FITC, PE and 7-AAD, the 
FITC that gets into FL3 will be essentailly removed when you sample Fl2
(PE only) and remove it from FL3 (7-AAD). If you are using *only* FITC and 7-AAD
you still need to setup FL2 in order to remove the FITC from FL3. In
this case you run FITC but use FL3 - FL2 to compensate it from FL3, even though
you will use FL1 to measure it. 

			Hope this helps,

			Joe



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