Measurement of ATP concentrations in mitochondria of living cells using luminescence and fluorescence approaches

Methods Cell Biol. 2020:155:199-219. doi: 10.1016/bs.mcb.2019.10.007. Epub 2019 Nov 27.

Abstract

Adenosine 5'-triphosphate (ATP) is the central metabolite in the energy metabolism of cells and is hydrolyzed to ADP and inorganic phosphate to provide free energy in various cellular processes. ATP also functions as an intracellular signaling molecule. Thus, it is important to know the ATP concentration within cells to understand cellular activities. Here, we describe two methods to detect ATP concentrations in the cytoplasm and mitochondrial matrix using genetically encoded luminescent or fluorescent biosensors. These methods enable quantitative investigation of ATP concentration dynamics in living cells, single cells and cell populations.

Keywords: ATP; Bioluminescence; Biosensor; FRET; Fluorescence; Live cell imaging; Luciferase assay; Mitochondria.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / analysis*
  • Animals
  • Biosensing Techniques / methods*
  • Cell Survival
  • Firefly Luciferin / metabolism
  • Fluorescence
  • HeLa Cells
  • Humans
  • Luciferases / metabolism
  • Luminescence*
  • Mitochondria / metabolism*

Substances

  • Firefly Luciferin
  • Adenosine Triphosphate
  • Luciferases