Abstract
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Yousif LF et al (2009) Targeting mitochondria with organelle-specific compounds: strategies and applications. Chembiochem 10:1939–1950
Johnson LV et al (1980) Localization of mitochondria in living cells with rhodamine 123. Proc Natl Acad Sci U S A 77:990–994
Ehrenberg B et al (1988) Membrane potential can be determined in individual cells from the nernstian distribution of cationic dyes. Biophys J 53:785–794
Johnson LV et al (1981) Monitoring of relative mitochondrial membrane potential in living cells by fluorescence microscopy. J Cell Biol 88:526–535
Reers M et al (1991) J-aggregate formation of a carbocyanine as a quantitative fluorescent indicator of membrane potential. Biochemistry 30:4480–4486
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 73:220–228
Rottenberg H, Wu S (1998) Quantitative assay by flow cytometry of the mitochondrial membrane potential in intact cells. Biochim Biophys Acta 1404:393–404
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 182:367–377
Acknowledgment
J.C.C. research is supported by Parkinson’s UK grant G-1101.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media New York
About this protocol
Cite this protocol
Corona, J.C., Duchen, M.R. (2014). Mitochondrial Bioenergetics Assessed by Functional Fluorescence Dyes. In: Hirrlinger, J., Waagepetersen, H. (eds) Brain Energy Metabolism. Neuromethods, vol 90. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1059-5_7
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1059-5_7
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1058-8
Online ISBN: 978-1-4939-1059-5
eBook Packages: Springer Protocols