Apoptosis with PI

From: Manickam Janakiraman (manickamj@mail.uni-wuerzburg.de)
Date: Sat Feb 23 2002 - 09:00:28 EST


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. I analysed the dot
plots and i could see the difference in apoptotic and live cell
population clearly. So i tried adjusting all the instrument settings
possible but i could not get rid of it.
                            Does someone have this experience ? If any
explanation or suggestion to overcome this problem is appreciated. As no
people around me could help me out i am in need of some answer from the
group. Thanks in advance.

Sincerely yours,

Manickam Janakiraman

Bayerische Julius Maximilians
University of Wuerzburg,
Institute for Medical Radiology
and cell Research,(MSZ),
Versbacher Str. 5,
D-97078 Wuerzburg,
Germany
Phone:  (49)-931-201-3838 (Lab)
             (49)-931-201-8331 (Res)
Fax:      (49)-931-201-3835
email:     manickamj@mail.uni-wuerzburg.de



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