I have tried using 7AAD for viability using the procedure from Current Protocols in Cytometry section 9.2.3. I made the stock solution using the procedure found on 7.11.4. I am using a FacsCalibur flow routinely used for three color applications and selected the Percp setting for my analysis. The results on one bone marrow sample demostrated clear negative and positive populations, but included a moderately bright 7AAD positive population between the neg and pos population. The positive population was determined/verified by setting up a previously fixed sample, CDChex, as a positive control and the bone marrow sample used was tested on the same day it was collected. I would like to have some imput and a few alternate procedures to follow to eliminate the indeterminate populations or better understand the significance of the moderately bright 7AAD population on a fresh sample. Mel
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