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

2005

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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