Open AccessA single-wavelength flow cytometric approach using redox-sensitive green fluorescent protein probes for measuring redox stress in live cells
Author(s) -
Elizabeth R Denn,
Joseph Schober
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2020-0168
Subject(s) - oxidative stress , chinese hamster ovary cell , redox , cytoplasm , green fluorescent protein , microbiology and biotechnology , mitochondrion , flow cytometry , biology , biophysics , oxidative phosphorylation , apoptosis , fluorescence , cell culture , chemistry , biochemistry , physics , organic chemistry , quantum mechanics , gene , genetics
Cellular redox changes are common in apoptosis, immune function, signaling pathways and cancer. The authors aimed to develop a single-wavelength method using the superior fluorescence sensitivity of a flow cytometer for measuring redox-sensitive green fluorescent protein signal during oxidative stress in cell lines. The single-wavelength method was able to discern small differences in oxidative stress between cell lines and between the cytoplasmic and mitochondrial compartments within the same cell line. In Chinese hamster ovary cells, the mitochondrial matrix compartment was more sensitive to oxidative stress compared with MDA-MB-231 cells, and the rapid changes in redox state were followed by a slow recovery phase. The authors conclude that this simplified method is useful and preferred for studies where alterations in overall redox-sensitive green fluorescent protein expression are controlled.