Re: Apoptosis with PI

From: Jáksó Pál (Pal.Jakso@aok.pte.hu)
Date: Tue Feb 26 2002 - 03:00:53 EST


Manickam Janakiraman wrote:

>
> Dear Flow users,
>
>                             I am new to the flowcytometry technique. I
> intend using it for the detection of apoptosis in mouse embryo
> fibroblasts. I tried some control and staurosporine treated cells in the
> FACSCalibur after staining with PI. Contrary to my expectation the DNA
> histograms looked different. There was a peak at hypodiploid position in
> the all the samples measured , even in control ones.

The hypodiplod sub-G1 peak is a natural sign of apoptosis if you stain
with PI after permeabilization.
You can control the unusual observation namely that your control sample
also shows a hypodiploid sub G1 peak is applying of other apoptosis
detecting methods along the  PI.
Otherwise are you sure that your control cells does not exhibit
apoptosis ?

> I analysed the dot
> plots and i could see the difference in apoptotic and live cell
> population clearly.

What kind of dot plots do you use?
Could you present some dot plots and PI histograms copying or attaching
them  into an email?



Best regards,

Pal Jakso

University of Pecs
Faculty of Medicine
Dept. of Pathology
7643, Hungary
12. Szigeti str.



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