Mitochondrial Bioenergetics Assessed by Functional Fluorescence Dyes

  • Juan Carlos Corona
  • Michael R. DuchenEmail author
Part of the Neuromethods book series (NM, volume 90)


Mitochondria play key roles in physiology and in disease. Understanding the mechanisms that define mitochondrial function, the regulation of energy balance in the cell, and the ways in which mitochondrial dysfunction impacts on cell physiology represents a key challenge in modern cellular pathophysiology. This requires a study of mitochondrial function in relation to the physiology of the cell, and so has driven approaches to measure and characterize key aspects of mitochondrial function within living cells. Central to this advance has been the use of fluorescent indicators that report mitochondrial membrane potential, mitochondrial redox state, rates of free radical generation, ATP, and so on. In this chapter we provide a critical appraisal and guide to the measurement of mitochondrial bioenergetics within living cells and discuss both the strengths and potential pitfalls of the most useful probes for mitochondrial membrane potential using confocal microscopy and flow cytometry.

Key words

Mitochondria Membrane potential Cell imaging Tetramethylrhodamine methyl ester Rhodamine 123 Confocal microscopy Flow cytometry 



J.C.C. research is supported by Parkinson’s UK grant G-1101.


  1. 1.
    Yousif LF et al (2009) Targeting mitochondria with organelle-specific compounds: strategies and applications. Chembiochem 10:1939–1950PubMedCrossRefGoogle Scholar
  2. 2.
    Johnson LV et al (1980) Localization of mitochondria in living cells with rhodamine 123. Proc Natl Acad Sci U S A 77:990–994PubMedCrossRefPubMedCentralGoogle Scholar
  3. 3.
    Ehrenberg B et al (1988) Membrane potential can be determined in individual cells from the nernstian distribution of cationic dyes. Biophys J 53:785–794PubMedCrossRefPubMedCentralGoogle Scholar
  4. 4.
    Johnson LV et al (1981) Monitoring of relative mitochondrial membrane potential in living cells by fluorescence microscopy. J Cell Biol 88:526–535PubMedCrossRefGoogle Scholar
  5. 5.
    Reers M et al (1991) J-aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential. Biochemistry 30:4480–4486PubMedCrossRefGoogle Scholar
  6. 6.
    Chinopoulos C et al (1999) Depolarization of in situ mitochondria due to hydrogen peroxide-induced oxidative stress in nerve terminals: inhibition of alpha-ketoglutarate dehydrogenase. J Neurochem 73:220–228PubMedCrossRefGoogle Scholar
  7. 7.
    Rottenberg H, Wu S (1998) Quantitative assay by flow cytometry of the mitochondrial membrane potential in intact cells. Biochim Biophys Acta 1404:393–404PubMedCrossRefGoogle Scholar
  8. 8.
    Zamzami N et al (1995) Sequential reduction of mitochondrial transmembrane potential and generation of reactive oxygen species in early programmed cell death. J Exp Med 182:367–377PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Laboratory of Neurosciences“Federico Gomez” Children’s Hospital of MexicoMexico CityMexico
  2. 2.Department of Cell and Developmental BiologyUniversity College LondonLondonUK

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