Use of flow cytometry as a tool to study mitochondrial membrane potential in isolated, living hepatocytes.
The present paper describes the possibility of determination of mitochondrial membrane potential (Deltapsi) in isolated hepatocytes making use of a Deltapsi-sensitive dye, i.e., the lipophilic cationic probe 5,5',6,6'-tetrachloro-1,1',3, 3'-tetraethylbenzimidazolcarbocyanine iodide (JC-1) and of cytofluorimetry. The validity of the method was proved by treating hepatocytes with FCCP (decrease of Deltapsi) and subsequent addition of 6-ketocholestanol (increase of Deltapsi). The results indicate that the proposed method may be used in laboratory practice.
Salvioli
S
S
Department of Biomedical Sciences, Section of General Pathology, University of Modena, via Campi, 287, 41100 Modena, Italy. bobyleva@unimo.it
Maseroli
R
R
Pazienza
T L
TL
Bobyleva
V
V
Cossarizza
A
A
D000818:Animals; D001562:Benzimidazoles / Q000378:metabolism; D002232:Carbocyanines / Q000378:metabolism; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D066298:In Vitro Techniques; D007425:Intracellular Membranes / Q000187:drug effects / Q000502:physiology; D007653:Ketocholesterols / Q000494:pharmacology; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D008297:Male; D008564:Membrane Potentials / Q000187:drug effects; D008928:Mitochondria / Q000187:drug effects / Q000502:physiology*; D010101:Oxygen Consumption / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D014475:Uncoupling Agents / Q000494:pharmacology
D001562:Benzimidazoles; D002232:Carbocyanines; D005456:Fluorescent Dyes; D007653:Ketocholesterols; D014475:Uncoupling Agents; C068624:5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolocarbocyanine; C063833:6-ketocholestanol; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone