Bare nuclei will stain PI+ve, Annexin V-ve. If you have access to s sorter,
I would sort a few onto a microcope slide and have a look at them.
Otherwise if look at some samples under a fluorescence microscope, you can
check if there are any bare nuclei.
Cells in an advanced stage of necrosis will have a lower DNA content
(because the DNA will be heavily degraded. I would expect to find them in
the Annexin +ve populatin with an intermediate PI staining.
Michael Ormerod
Message text written by Martin Kelly
>Dear friends,
we would like to ask some advice r.e. the interpretation of some
work we have been doing on induced sputum and BAL from normal
subjects and COPD patients. We have been using 'flow' to look at
the granulocytes, and examine the AV and PI staining in the
events which we have gated on, which we feel are in the region
in which one would expect to find granulocytes. Cells coming up
AV+PI- we interpret as apoptotic; AV+PI+ as necrotic. This would
seem to be conventionally how it is desribed in papers we have
looked at. However, we were not sure how one might interpret
cells which are AV-PI+? Are these just cells in an advanced
state of necrosis? Is there another interpretation?
We value any advice.
With best wishes. yours sincerely,
Martin Kelly<
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