re: Re: PI vs. 7-AAD Nuclear Staining
C.Reutelingsperger@BIOCH.RULIMBURG.NL
Mon, 18 Mar 1996 15:47:05 +0100 (MET)
For those of you, who are using Annexin V-FITC and propidium iodide to measure apoptosis and have
problems with dim PI positivity of viable and early apoptotic cells, you can wash your cells after
staining with Annexin V-FITC and PI.
Using the binding buffer coming with the APOPTEST-FITC kit of NeXins Research, the washing step will
not result in a significant decrease of cellular bound Annexin V-FITC.
Doing this you will miss a possibly interesting subpopulation, namely those cells that are about to enter
the secondary necrotic subpopulation. Those cells start to lose plasma membrane integrity without
having a fully lost barrier (a free passage of PI to the intracellular DNA) and represent a late phase of
apoptosis.
So, it'll depend on your question whether or not to include a washing step following Annexin V-FITC and
propidium iodide staining.
Chris Reutelingsperger
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
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If you have any comments please direct them to Dr. J. Paul Robinson, Professor & Director, PUCL,
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EMAIL robinson@flowcyt.cyto.purdue.edu
