Method for live-cell super-resolution imaging of mitochondrial cristae and quantification of submitochondrial membrane potentials

Dane M Wolf; Mayuko Segawa; Orian S Shirihai; Marc Liesa

doi:10.1016/bs.mcb.2019.12.006

Method for live-cell super-resolution imaging of mitochondrial cristae and quantification of submitochondrial membrane potentials

Methods Cell Biol. 2020:155:545-555. doi: 10.1016/bs.mcb.2019.12.006. Epub 2020 Mar 4.

Authors

Dane M Wolf¹, Mayuko Segawa², Orian S Shirihai³, Marc Liesa⁴

Affiliations

¹ Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, United States; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, United States; Graduate Program in Nutrition and Metabolism, Graduate Medical Sciences, Boston University School of Medicine, Boston, MA, United States.
² Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, United States; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, United States.
³ Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, United States; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, United States. Electronic address: oshirihai@mednet.ucla.edu.
⁴ Department of Medicine, Endocrinology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, United States; Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA, United States; Molecular Biology Institute, University of California Los Angeles, Los Angeles, CA, United States. Electronic address: mliesa@mednet.ucla.edu.

Abstract

The emergence of diffraction-unlimited live-cell imaging technologies has enabled the examination of mitochondrial form and function in unprecedented detail. We recently developed an approach for visualizing the inner mitochondrial membrane and determined that cristae membranes possess distinct mitochondrial membrane potentials, representing unique bioenergetic subdomains within the same organelle. Here, we outline a methodology for resolving cristae and inner boundary membranes using the LSM880 with Airyscan. Furthermore, we demonstrate how to analyze TMRE fluorescence intensity using the Nernst equation to calculate membrane potentials of individual cristae. Altogether, using these new techniques to study the electrochemical properties of the cristae can help to gain deeper insight into the still elusive nature of the mitochondrion.

Keywords: Airyscan; Cristae; Live-cell imaging; Mitochondrial membrane potential; Super-resolution imaging.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Cell Survival
HeLa Cells
Humans
Imaging, Three-Dimensional / methods*
Membrane Potential, Mitochondrial*
Mitochondria / metabolism*
Mitochondrial Membranes / metabolism*

Abstract

Publication types

MeSH terms

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