Re: FACScalibur instrument settings for PI ploidy/apoptosis acquisition

From: Tim Kute (tkute@wfubmc.edu)
Date: Wed Aug 01 2001 - 08:59:29 EST


In most cases, I like to have my G1 peak start  in the 1/3 rd of the x axis
range.  If you are looking for apoptotic cells, you might want it higher
around 2/3 rd of range.  If you really want to see every event, you might
even want to put the scale on log and have the G1 peak at the 80% area of
the x axis.

Tim Kute

Maciej Simm wrote:

> I'm posting this on behalf of a collegue. She's doing an assay with
> just PI looking at the peaks before/after incubation of her cells in
> pro-apoptotic drugs.  If anyone looks at PI on a Calibur, could you
> fill me in on the instrument settings that work the best? I would
> appreciate any other tips.
>
> Maciej
>
> >find out voltage settings for the PI assay. in particular ask  about
> >a printout of an acquisition template they use. also -
>
> >fsc    lin or log? what voltage?
> >ssc   same as above
> >fsc    s.a.a.
> >fl2     s.a.a.
> >fl3     s.a.a.
>
> >what about compensation?
>
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