This paper presents a high-content microscopy workflow for simultaneous quantification of intracellular ROS levels, as well as mitochondrial membrane potential and morphology – jointly referred to as mitochondrial morphofunction – in living adherent cells using the cell-permeant fluorescent reporter molecules 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H2DCFDA) and tetramethylrhodamine methylester (TMRM).
Sieprath, T., Corne, T., Robijns, J., Koopman, W. J. H., De Vos, W. H. Cellular Redox Profiling Using High-content Microscopy. J. Vis. Exp. (123), e55449, doi:10.3791/55449 (2017).