A Plate Reader-Based Measurement of the Cellular ROS Production Using Dihydroethidium and MitoSOX

Chih-Yao Chung; Michael R Duchen

doi:10.1007/978-1-0716-2309-1_24

A Plate Reader-Based Measurement of the Cellular ROS Production Using Dihydroethidium and MitoSOX

Methods Mol Biol. 2022:2497:333-337. doi: 10.1007/978-1-0716-2309-1_24.

Authors

Chih-Yao Chung¹, Michael R Duchen²

Affiliations

¹ Department of Cell and Developmental Biology and Consortium for Mitochondrial Research, UCL, London, UK.
² Department of Cell and Developmental Biology and Consortium for Mitochondrial Research, UCL, London, UK. m.duchen@ucl.ac.uk.

PMID: 35771455
DOI: 10.1007/978-1-0716-2309-1_24

Abstract

Intracellular reactive oxygen species (ROS) act as an important signaling transductor in cells, regulating almost every aspect of cell biology. Measurements of ROS production thus, offer links between oxidative stress and cell pathophysiology. Here, we describe a simple screening assay in intact adherent cells by fluorescence microplate readers, using dihydroethidium (DHE) and MitoSOX to measure cytosolic superoxide and mitochondrial superoxide production, respectively. This assay enables a quick and reliable assessment of ROS generation in a well-controlled environment.

Keywords: Dihydroethidium; MitoSOX; Mitochondria; Plate reader; Reactive oxygen species.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Ethidium / analogs & derivatives
Oxidative Stress*
Phenanthridines
Reactive Oxygen Species* / analysis
Superoxides* / analysis

Substances

5-(6'-triphenylphosphoniumhexyl)-5,6-dihydro-6-phenyl-3,8-phenanthridinediammine
Phenanthridines
Reactive Oxygen Species
dihydroethidium
Superoxides
Ethidium