Analysis of the membrane potential of rat- and mouse-liver mitochondria by flow cytometry and possible applications

Our results suggest that flow-cytometry analysis of small mitochondrial samples stained with membrane-potential-sensitive probes could be a valuable tool offering fast experimental procedure with extremely small samples, to study the mitochondrial behaviour in pathological situations affecting the mitochondrial population within the cell

Patrice X. PETIT


Scholarcy highlights

  • Washed and purified rat- or mouse-liver mitochondria exhibiting high membrane integrity and metabolic activity were studied by flow cytometry
  • In order to minimize the non-specific binding of dyes on nonmitochondrial membranes during the flow-cytometric studies, rat-liver mitochondria were purified on a discontinuous Percoll gradient to give a purified mitochondrial fraction collected at the 30/70% interface
  • These preparations of washed and purified mitochondria were characterized in terms of membrane integrity, membrane potential, and oxidative and phosphorylative properties
  • Frequency the substrate and collapse of A y by valinomycin. These results show that, as expected from fluorescence of dyes in the bulk population of mitochondria [9, 26, 421, safranine 0,rhodamine 123 and 3,3'-dihexyloxadicarbocyanine iodide behave as potentiometric probes in flow cytometry
  • Only fluorescence probes are normally useful for flow cytometry
  • High probe concentrations may act as uncouplers [9, 50, 511 and/ or lead to inner quenching; low probe concentrations avoid these problems, provided the fluorescence signal is adequate

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