Protocol for Imaging of Mitoflashes in Live Cardiomyocytes

STAR Protoc. 2020 Sep 3;1(2):100101. doi: 10.1016/j.xpro.2020.100101. eCollection 2020 Sep 18.

Abstract

We describe a protocol for imaging a mitochondrial fluorescence transient increase event (Mitoflash) in live cardiomyocytes using a confocal microscope. Mitoflash, detected by mitochondria-targeted circularly permuted fluorescent protein (mt-cpYFP), can be used to assess mitochondrial respiration function in situ. The protocol is also suitable for live-cell imaging of other adherent cells, including fibroblasts and hepatocytes. For complete details on the use and execution of this protocol, please refer to Gong et al. (2014) and Gong et al. (2015).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Respiration / physiology
  • Fluorescence
  • Humans
  • Luminescent Proteins / metabolism
  • Microscopy, Confocal / methods
  • Mitochondria / metabolism
  • Mitochondria / physiology*
  • Muscle, Skeletal / metabolism
  • Myocytes, Cardiac / metabolism
  • Myocytes, Cardiac / physiology*
  • Optical Imaging / methods*

Substances

  • Luminescent Proteins