Abstract
The cross talk between mitochondrial dynamic structure, determined primarily by mitochondrial fission and fusion events, and mitochondrial function of energetics, primarily ATP and ROS production, is widely appreciated. Understanding the mechanistic details of such cross talk between mitochondrial structure and function needs integrated quantitative analyses between mitochondrial dynamics and energetics. Here we describe our recently designed approach of mito-SinCe2 that involves high resolution confocal microscopy of genetically expressed ratiometric fluorescent probes targeted to mitochondria, and its quantitative analyses. Mito-SinCe2 analyses allows for quantitative analyses of mitochondrial structure–function relationship in single cells toward understanding the role of mitochondria and their heterogeneity in various physiological and pathological conditions.
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Acknowledgments
We acknowledge Drs. S. Rafelski and M. Vienna for developing the MitoGraph v2.1 software and for help with accessing and understanding its capabilities; Dr. P. Gupta for helping to develop the MitoSinCe2 approach in its earliest stages; D. Parker for performing some of the experiments from which we took example data; and J. Wirth for the custom-built Excel macro. This work was supported by the National Institutes of Health (NIH) [R33ES025662] grant to B.S. and K.M.
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Spurlock, B., Mitra, K. (2021). Mito-SinCe2 Approach to Analyze Mitochondrial Structure–Function Relationship in Single Cells. 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_27
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DOI: https://doi.org/10.1007/978-1-0716-1262-0_27
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