title: High Throughput-Based Mitochondrial Function Assays by Multi-Parametric Flow Cytometry.
abstract: Mitochondrial dysfunction has been increasingly implicated as an important mechanism for chemical-induced toxicity. In the present unit, we describe a multi-parametric flow cytometry assay to assess the effects of drug or chemical-induced mitochondrial dysfunction in cells. Cells are cultured in a glucose-supplemented medium and exposed to increasing concentrations of various chemicals. Several key mitochondrial/cellular parameters known to be directly impacted by mitochondrial dysfunction, such as mitochondrial membrane potential (MMP), mitochondrial reactive oxygen species (ROS) production, intracellular reduced glutathione (GSH) level, and cell viability, are simultaneously measured by flow cytometry.
authors.affiliation: Purdue University Cytometry Laboratories, West Lafayette, Indiana.
authors.forename: J Paul
authors.initials: JP
authors.lastname: Robinson
authors.affiliation: Amgen, Thousand Oaks, California.
authors.forename: Nianyu
authors.initials: N
authors.lastname: Li
authors.affiliation: Amgen, Thousand Oaks, California.
authors.forename: Padma Kumar
authors.initials: PK
authors.lastname: Narayanan
mesh_terms: D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000378:metabolism; D018922:HL-60 Cells; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D013194:Staining and Labeling
chemical_list: D005456:Fluorescent Dyes
keywords: cell viability; flow cytometry; mitochondria; mitochondrial dysfunction; mitochondrial membrane potential; reactive oxygen species (ROS)