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Live-Cell Assessment of Reactive Oxygen Species Levels Using Dihydroethidine

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Mitochondrial Medicine

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2275))

Abstract

Reactive oxygen species (ROS) play an important role in cellular (patho)physiology. Empirical evidence suggests that mitochondria are an important source of ROS, especially under pathological conditions. Here, we describe a method for ROS measurement using dihydroethidium (HEt) and live-cell microscopy.

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Abbreviations

FCCP:

carbonyl cyanide-p-trifluoromethoxyphenylhydrazone

HEt:

dihydroethidium

HT:

HEPES-Tris

mito-HEt:

mito-dihydroethidium

ROS:

reactive oxygen species

TPP:

triphenylphosphonium

Δψ :

mitochondrial membrane potential

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Acknowledgments

We are grateful to Dr. A.S. De Jong and Dr. Marleen Forkink (Dept. of Biochemistry, Radboudumc) for performing the HEt and Mito-HEt experiments.

Potential competing interests: Werner J.H. Koopman is a scientific advisor of Khondrion. This SME had no involvement in the data collection, analysis, and interpretation; writing of the manuscript; and the decision to submit the manuscript for publication.

Author information

Authors and Affiliations

  1. Human and Animal Physiology, Wageningen University & Research, Wageningen, Netherlands

    Sander Grefte & Werner J. H. Koopman

  2. Department of Biochemistry, Radboud Institute for Molecular Life Sciences, Radboud Center for Mitochondrial Medicine, Radboudumc, Nijmegen, The Netherlands

    Werner J. H. Koopman

Authors
  1. Sander Grefte
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  2. Werner J. H. Koopman
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Corresponding author

Correspondence to Werner J. H. Koopman .

Editor information

Editors and Affiliations

  1. Department of Pharmaceutical Sciences, Midwestern University, College of Pharmacy, Glendale, AZ, USA

    Volkmar Weissig

  2. Cochin Hospital, Cochin Institute, INSERM U1016, Paris, France

    Marvin Edeas

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Grefte, S., Koopman, W.J.H. (2021). Live-Cell Assessment of Reactive Oxygen Species Levels Using Dihydroethidine . In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine . Methods in Molecular Biology, vol 2275. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1262-0_18

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  • DOI: https://doi.org/10.1007/978-1-0716-1262-0_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1261-3

  • Online ISBN: 978-1-0716-1262-0

  • eBook Packages: Springer Protocols

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