Calibration and measurement of mitochondrial pH in intact adult rat cardiomyocytes

STAR Protoc. 2021 May 14;2(2):100543. doi: 10.1016/j.xpro.2021.100543. eCollection 2021 Jun 18.

Abstract

Mitochondrial pH is a vital parameter of the mitochondrial environment, which determines the rate of many mitochondrial functions, including metabolism, membrane potential, fate, etc. Abnormal mitochondrial pH is always closely related to the health status of cells. Analyzing mitochondrial pH can serve as a proxy for mitochondrial and cellular function. This protocol describes the use of SNARF-1 AM, a pH-sensitive fluorophore, to measure mitochondrial pH. This protocol details the steps to evaluate mitochondrial pH in live adult cardiomyocytes using confocal microscopy. The protocol can be adapted to other adherent cell types. For complete details on the use and execution of this protocol, please refer to Wei-LaPierre et al. (2013).

Keywords: Cell Biology; Metabolism; Microscopy; Molecular/Chemical Probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Benzopyrans / analysis
  • Benzopyrans / chemistry
  • Cytological Techniques / methods*
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / chemistry
  • Hydrogen-Ion Concentration*
  • Mitochondria* / chemistry
  • Mitochondria* / metabolism
  • Myocytes, Cardiac / chemistry*
  • Myocytes, Cardiac / cytology
  • Naphthols / analysis
  • Naphthols / chemistry
  • Rats
  • Rhodamines / analysis
  • Rhodamines / chemistry

Substances

  • Benzopyrans
  • Fluorescent Dyes
  • Naphthols
  • Rhodamines
  • seminaphthorhodaminefluoride