Viability with PI

From: Cheryl Smith (cheryla.smith@utoronto.ca)
Date: Tue Feb 13 2001 - 08:34:34 EST


Hello All,

A couple last questions about PI staining for viability.

I have found that very often there is a population of lower intensity PI as
well as the very high one that we typically define as dead.  I think there
are logical explanations for the presence of this, but the problem is how
can you get away without using a dedicated detector for PI and be sure that
some of what you are assuming to be, for instance, positive PE or PE/CY5 is
not actually low intensity PI?

I have also seen, even with preparations that have been washed after the
addition of PI, an increase in the background of the PE signal. I'm not
really sure why this happens, but do people just ignore it?  Wouldn't it
cause as much problem with samples of not extremely bright, and relatively
rare, positive populations as would a few, maybe less than 1%,
non-specifically staining dead cells?  And you can't really compensate it
away.

These things have nagged me for a long time. I hope someone can offer some
advice.

Thanks,
Cheryl



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