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@uz.kuleuven.ac.be -------------------------------------
This archive was generated by hypermail 2b29 : Sat Mar 10 2001 - 19:31:22 EST