Re: Viability assay using flow cytometry

From: Glenn Paradis (gap@MIT.EDU)
Date: Fri Apr 28 2000 - 17:21:51 EST


Hi Steve,

Here is a quick response to compensation issue between Fl1 and Fl3 on a
Vantage.  Just take the signal lead (not the black voltage lead) off the
Fl3 PMT and put the signal lead from the Fl2 PMT on your Fl3 PMT.  Your FL3
data will be routed to your Fl2 electronics and thus you can now do Fl2-%
Fl1 compensation.  To display your data put up a dot plot of Fl1 vs Fl2.

Take care.

Glenn
MIT


>Dear flowers,
>
>What is the best way to perform a viability assay using flow cytometry? I
>am currently using calcein AM and propidium iodide (PI). I substituted the
>ethidium homodimer with PI only because PI appears to be a common indicator
>of dead cells in flow cytometry and it is less expensive. For those of you
>who have FACS Vantages, is it possible to compensate between FL1( Calcein)
>and FL3 ( PI) in your systems?
>
>For some reason, I do not see distinct populations of cells when I use the
>previously mentioned stains. In fact, I see a population that appears to be
>positive for PI and positive for calcein based on controls ( live cells:
>calcein only and Etoh fixed cells: PI only).
>All responses are appreciated.
>
>Steve



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