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J Cell Biol. 1981 March 1; 88(3): 526–535.
PMCID: PMC2112765

Monitoring of relative mitochondrial membrane potential in living cells by fluorescence microscopy


Permeant cationic fluorescent probes are shown to be selectively accumulated by the mitochondria of living cells. Mitochondria-specific interaction of such molecules is apparently dependent on the high trans- membrane potential (inside negative) maintained by functional mitochondria. Dissipation of the mitochondrial trans-membrane and potential by ionophores or inhibitors of electron transport eliminates the selective mitochondrial association of these compounds. The application of such potential-dependent probes in conjunction with fluorescence microscopy allows the monitoring of mitochondrial membrane potential in individual living cells. Marked elevations in mitochondria- associated probe fluorescence have been observed in cells engaged in active movement. This approach to the analysis of mitochondrial membrane potential should be of value in future investigations of the control of energy metabolism and energy requirements of specific biological functions at the cellular level.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Akerman KE, Wikström MK. Safranine as a probe of the mitochondrial membrane potential. FEBS Lett. 1976 Oct 1;68(2):191–197. [PubMed]
  • Azzi A, Chance B, Radda GK, Lee CP. A fluorescence probe of energy-dependent structure changes in fragmented membranes. Proc Natl Acad Sci U S A. 1969 Feb;62(2):612–619. [PubMed]
  • Bakeeva LE, Grinius LL, Jasaitis AA, Kuliene VV, Levitsky DO, Liberman EA, Severina II, Skulachev VP. Conversion of biomembrane-produced energy into electric form. II. Intact mitochondria. Biochim Biophys Acta. 1970 Aug 4;216(1):13–21. [PubMed]
  • Bashford CL, Smith JC. The use of optical probes to monitor membrane potential. Methods Enzymol. 1979;55:569–586. [PubMed]
  • Cohen LB, Salzberg BM. Optical measurement of membrane potential. Rev Physiol Biochem Pharmacol. 1978;83:35–88. [PubMed]
  • Colnna R, Massari S, Azzone GF. The problem of cation-binding sites in the energized membrane of intact mitochondria. Eur J Biochem. 1973 May 2;34(3):577–585. [PubMed]
  • Grinius LL, Jasaitis AA, Kadziauskas YP, Liberman EA, Skulachev VP, Topali VP, Tsofina LM, Vladimirova MA. Conversion of biomembrane-produced energy into electric form. I. Submitochondrial particles. Biochim Biophys Acta. 1970 Aug 4;216(1):1–12. [PubMed]
  • Hladky SB, Rink TJ. Potential difference and the distribution of ions across the human red blood cell membrane; a study of the mechanism by which the fluorescent cation, diS-C3-(5) reports membrane potential. J Physiol. 1976 Dec;263(2):287–319. [PubMed]
  • Hoffman JF, Laris PC. Determination of membrane potentials in human and Amphiuma red blood cells by means of fluorescent probe. J Physiol. 1974 Jun;239(3):519–552. [PubMed]
  • Johnson LV, Walsh ML, Chen LB. Localization of mitochondria in living cells with rhodamine 123. Proc Natl Acad Sci U S A. 1980 Feb;77(2):990–994. [PubMed]
  • Lampidis TJ, Schaiberger GE. Age-related loss of DNA repair synthesis in isolated rat myocardial cells. Exp Cell Res. 1975 Dec;96(2):412–416. [PubMed]
  • Laris PC, Bahr DP, Chaffee RR. Membrane potentials in mitochondrial preparations as measured by means of a cyanine dye. Biochim Biophys Acta. 1975 Mar 20;376(3):415–425. [PubMed]
  • McClure WO, Edelman GM. Fluorescent probes for conformational states of proteins. II. The binding of 2-p-toluidinylnaphthalene-6-sulfonate to alpha-chymotrypsin. Biochemistry. 1967 Feb;6(2):559–566. [PubMed]
  • Shapiro HM, Natale PJ, Kamentsky LA. Estimation of membrane potentials of individual lymphocytes by flow cytometry. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5728–5730. [PubMed]
  • Sims PJ, Waggoner AS, Wang CH, Hoffman JF. Studies on the mechanism by which cyanine dyes measure membrane potential in red blood cells and phosphatidylcholine vesicles. Biochemistry. 1974 Jul 30;13(16):3315–3330. [PubMed]
  • Stryer L. The interaction of a naphthalene dye with apomyoglobin and apohemoglobin. A fluorescent probe of non-polar binding sites. J Mol Biol. 1965 Sep;13(2):482–495. [PubMed]
  • Summerhayes IC, Franks LM. Effects of donor age on neoplastic transformation of adult mouse bladder epithelium in vitro. J Natl Cancer Inst. 1979 Apr;62(4):1017–1023. [PubMed]
  • Waggoner A. Optical probes of membrane potential. J Membr Biol. 1976 Jun 30;27(4):317–334. [PubMed]
  • Waggoner AS. Dye indicators of membrane potential. Annu Rev Biophys Bioeng. 1979;8:47–68. [PubMed]
  • Waggoner AS. The use of cyanine dyes for the determination of membrane potentials in cells, organelles, and vesicles. Methods Enzymol. 1979;55:689–695. [PubMed]
  • Kinnally KW, Tedeschi H, Maloff BL. Use of dyes to estimate the electrical potential of the mitochondrial membrane. Biochemistry. 1978 Aug 8;17(16):3419–3428. [PubMed]

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