Detection of Mitochondrial Mass, Damage, and Reactive Oxygen Species by Flow Cytometry

Cold Spring Harbor Protocols
Daniel Puleston

Abstract

The reagents and procedures highlighted here will give the investigators an indication of the health status and volume of mitochondria in primary cells and cell lines through the use of a number of cell-permeable dyes. Mitochondrial volume can be monitored by using the probe MitoTracker Green FM. This reagent labels mitochondria in a manner that is independent of the membrane potential, therefore providing a readout relating purely to the mitochondrial mass of the cell. In contrast, MitoTracker Red CMXRos, tetramethylrhodamine methyl ester, and 10-N-nonyl acridine orange label mitochondria in a manner dependent on the membrane potential, thus giving an indication of mitochondrial stress. Using MitoSOX Red, it is also possible to analyze the production of the mitochondrial superoxide anion. Like the MitoTracker probes, MitoSOX Red is taken up passively by cells. In the mitochondria, the probe is oxidized by superoxide, resulting in the emission of red fluorescence.

References

Sep 4, 2015·Cold Spring Harbor Protocols·Daniel PulestonAnna Katharina Simon

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Citations

May 9, 2018·Biochemistry. Biokhimii︠a︡·A G RogovR A Zvyagilskaya
Aug 15, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Natália Aydos MarcondesFélix Hilário Díaz González
Nov 11, 2017·Cell Death and Differentiation·Lei SunXiaoqiang Yao
Jan 11, 2018·Science Signaling·Ji-Hoon LeeTanya T Paull
Jun 10, 2019·Journal of Alzheimer's Disease : JAD·John Z CavendishJames W Simpkins
Jun 16, 2017·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Xianghong ZhangMatthew R Rosengart
Jul 9, 2020·Journal of Cellular and Molecular Medicine·Tien-Hung HuangFan-Yen Lee
Jun 21, 2018·Current Protocols in Cell Biology·Dong Fu, Jennifer Lippincott-Schwartz
Jul 20, 2018·Nature Communications·Lele ZhuShao-Cong Sun
Mar 30, 2018·Frontiers in Cell and Developmental Biology·Shirisha ChittiboyinaSophie A Lelièvre
Oct 28, 2019·Cell Death & Disease·Sagnik Giri, Chandrima Shaha
Jun 20, 2020·MethodsX·Lauar de Brito MonteiroPedro M M Moraes-Vieira
Dec 11, 2020·Cell Death Discovery·José R Navarro-BetancourtAndrey V Cybulsky
Apr 3, 2020·Journal of Molecular and Cellular Cardiology·Aleksandr B StotlandSarah J Parker
Mar 23, 2021·Biotechnology and Bioengineering·Nadiya RomanovaThomas Noll
Aug 21, 2021·Journal of Analytical Methods in Chemistry·Vu Thi ThuNguyen Thi Ha Ly
Aug 28, 2021·International Journal of Molecular Sciences·Samuel AbokyiDennis Yan-Yin Tse
Sep 4, 2015·Cold Spring Harbor Protocols·Daniel PulestonAnna Katharina Simon

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