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Updated: Dec 26, 2025

Author Spotlight: Decoding Mitochondrial Aging
Published on: June 30, 2023
Method for live-cell super-resolution imaging of mitochondrial cristae and quantification of submitochondrial
Dane M Wolf1, Mayuko Segawa2, Orian S Shirihai2
1Department 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.
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.
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