Mireia Dalmau writes: > >We are studying the effect of some drugs on isolated mitochondria. >We are interested in determining how these drugs affect their >activity and membrane potential. >Does anyone know a good probe or protocol which could help us? We >have found a lot of cell permeant probes, but not a useful one to >distinguish active and inactive mitochondria in a cell-free system. > >We are using other techical approaches, but, if possible, we would >like to use flow cytometry. JC-1 should theoretically be usable as a ratiometric membrane potential probe in isolated mitochondria and bacteria; in my hands, it hasn't worked in bacteria and I haven't tried it in isolated mitochondria, because I don't work with isolated mitochondria. However, I think it is likely that the ratiometric method for bacterial membrane potential measurement recently described from my lab using diethyloxacarbocyanine (DiOC2(3)) (Novo D, Perlmutter NG, Hunt RH, Shapiro HM: Accurate flow cytometric membrane potential measurement in bacteria using diethyloxacarbocyanine and a ratiometric technique. Cytometry. 1999; 35:55-63) would work for isolated mitochondria. I understand that Molecular Probes now sells or will shortly be selling this dye. -Howard
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