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Are you setting the compensation values with SINGLE stained samples?
And yes, we all have seen the dim PI positives. I use as litte PI as possible
(0.5 micrograms per ml) but still have seen them. I've heard some people WASH
after PI staining, but the A-V protocols are supposed to be no wash!
Dennis
______________________________ Reply Separator _________________________________
Subject: PI vs. 7-AAD Nuclear Staining
Author: DJS@allp.com at @UCSD
Date: 3/15/96 10:24 AM
I have been trying to surface stain apopotic MCF-7 breast cancer cells with
Annexin V-FITC and stain the DNA with PI to test for necrosis vs apoptosis.
Even though the emission spectra shouldn't overlap I am having to
compensate for what appears to be overlap in both FL1 and FL3.
Also the PI appears to be passing through the membrane of viable cells.
Any suggestions on either problem (Compensation or viability stain)
Would switching to 7-AAD be a possibility.
Thanks for the input,
Dan Smith
San Diego
djs@allp.com
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