>From past experience of PMNs I'd say you have a high background from manipulation. Since this is in whole blood you could try a different anti-coagulant. I once collaborated with Alistair Crockard who was doing some work on CGD using DHR123 and flow cytometry. You could look up that paper and compare methods. In fact give me a day or so, I might have a copy, and be able to send you the methods. Craig Turner IBGRL ---------- From: helena.mareckova@lf1.cuni.cz To: cyto-inbox Subject: Oxidative Burst Test Date: Tuesday, March 14, 2000 21:10 We have recently started to look at oxidative burst as a means of assesing phagocytosis. We use whole-blood method with DHR123 as a substrate which is converting to rhodamine during oxidative burst. Our problem is in high spontaneous metabolic activity of PMN's: in the polypropylene tube with lithium-heparin whole blood + DHR123 +NaCl 0,9% we have after incubation and lysis too high intensity of fluorescence FL1. Does anyone have experiences with this problem? Thanks for your help. Karin Malickova kmali@lf1.cuni.cz
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