Hello flow cyt. users, I intend to measure intracellular H2O2 in vascular smooth muscle cells using H2DCFDA by flow cytometry. QUESTION 1: Does anyone know what conc. (nM or microM) of DCFDA the cells should be loaded with? I am not fixing the cells and I am carrying them to the flow cytometry facility (10 minutes walk), so I expect some leakage of the dye from the cells. I read that DHR123 is more sensitive, but I wish to measure net intracellular H2O2, rather than mitochondrial H2O2, hence the choice of H2DCFDA. QUESTION 2: Some researchers use fluorescence confocal microscopy for the same purpose? Are confocal and flow cyt. equally sensitive and accurate for this purpose? Thanks in advance, Kalyankrishna S. SKALYANKRISH@UTMEM1.UTMEM.EDU
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