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

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

7 Scopus citations

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.

Original languageEnglish
Title of host publicationMitochondria, 3rd Edition
EditorsLiza A. Pon, Eric A. Schon
PublisherAcademic Press Inc.
Pages545-555
Number of pages11
ISBN (Print)9780128202289
DOIs
StatePublished - 1 Jan 2020
Externally publishedYes

Publication series

NameMethods in Cell Biology
Volume155
ISSN (Print)0091-679X

Keywords

  • Airyscan
  • Cristae
  • Live-cell imaging
  • Mitochondrial membrane potential
  • Super-resolution imaging

ASJC Scopus subject areas

  • Cell Biology

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