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Evaluation of Mitochondria Content and Function in Live Cells by Multicolor Flow Cytometric Analysis

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Mitochondrial Medicine

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2276))

Abstract

To evaluate how a cell responds to the external stimuli, treatment, or alteration of the microenvironment, the quantity and quality of mitochondria are commonly used as readouts. However, it is challenging to apply mitochondrial analysis to the samples that are composed of mixed cell populations originating from tissues or when multiple cell populations are of interest, using methods such as Western blot, electron microscopy, or extracellular flux analysis.

Flow cytometry is a technique allowing the detection of individual cell status and its identity simultaneously when used in combination with surface markers. Here we describe how to combine mitochondria-specific dyes or the dyes targeting the superoxide produced by mitochondria with surface marker staining to measure the mitochondrial content and activity in live cells by flow cytometry. This method can be applied to all types of cells in suspension and is particularly useful for analysis of samples composed of heterogeneous cell populations.

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Acknowledgments

We would like to thank Dr. Chin-Wen Wei for the initial set up for this experimental system and Yu-Ting Hsieh for critically reading the manuscript. This work was supported by grants from Ministry of Science and Technology, Taiwan (MOST 107-2320-B-010-020, MOST 108-2628-B-010-005 to C.-L. H.; 107-2320-B-010 -016 -MY3, 106-2320-B-010 -026 -MY3 to I. L. D.) and Cancer Progression Research Center, National Yang-Ming University from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan.

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Authors and Affiliations

  1. Institute of Microbiology and Immunology, National Yang-Ming University, Taipei, Taiwan

    Hsiu-Han Fan, Tsung-Lin Tsai, Ivan L. Dzhagalov & Chia-Lin Hsu

Authors
  1. Hsiu-Han Fan
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  2. Tsung-Lin Tsai
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  3. Ivan L. Dzhagalov
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  4. Chia-Lin Hsu
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Corresponding author

Correspondence to Chia-Lin Hsu .

Editor information

Editors and Affiliations

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

    Volkmar Weissig

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

    Marvin Edeas

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Fan, HH., Tsai, TL., Dzhagalov, I.L., Hsu, CL. (2021). Evaluation of Mitochondria Content and Function in Live Cells by Multicolor Flow Cytometric Analysis. In: Weissig, V., Edeas, M. (eds) Mitochondrial Medicine . Methods in Molecular Biology, vol 2276. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-1266-8_15

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  • DOI: https://doi.org/10.1007/978-1-0716-1266-8_15

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  • Publisher Name: Springer, New York, NY

  • Print ISBN: 978-1-0716-1265-1

  • Online ISBN: 978-1-0716-1266-8

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