eosinophils, apoptosis and surface expression

Joeri Aerts joeri.aerts at uz.kuleuven.ac.be
Mon Jun 5 03:28:36 EST 2000


Dear all,

I am currently investigating the expression of several surface molecules on
eosinophils isolated from peripheral blood.  I am especially interested in
the evolution of expression due to apoptosis in these cells.  In order to
monitor apoptosis I use annexin-V-PE with 7-AAD, both from Pharmingen.  The
problem is that eos exhibit significant autofluorescence (which has been
used for gating).  This of course makes it difficult to tell what is due to
the MoAbs and what is due to the autofluorescence, especially for markers
that show a low median fluorescence such as CD69.  This can be managed for
eos that are in culture for about 24 hours, but from 48 hours onwards, the
autofluorescence seems to increase further which necessitates further
meddling with the settings and more importantly makes it difficult to
compare different time points.  Does anybody have  any experience with this?

Thanks a lot,

Joeri
-------------------------------------
Joeri Aerts

Experimental Laboratory Medicine -
Laboratory Medicine (Haematology)

CDG 7e VD - Gasthuisberg
Herestraat 49
B-3000 Leuven
BELGIUM

tel.:	+32-16347012
fax.:	+32-16347042
e-mail: joeri.aerts at uz.kuleuven.ac.be
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