title: A microfluidic systems biology approach for live single-cell mitochondrial ROS imaging.
abstract: Most current studies of reactive oxygen species (ROS) production report globally averaged measurements within the cell; however, ROS can be produced in distinct subcellular locations and have local effects in their immediate vicinity. A microfluidic platform for high-throughput single-cell imaging allows mitochondrial ROS production to be monitored as varying in both space and time. Using this systems biology approach, single-cell variability can be viewed within a population. We discuss single-cell monitoring of contributors to mitochondrial redox state-mitochondrial hydrogen peroxide or superoxide-through the use of a small molecule probe or targeted fluorescent reporter protein. Jurkat T lymphoma cells were stimulated with antimycin A and imaged in an arrayed microfluidic device over time. Differences in single-cell responses were observed as a function of both inhibitor concentration and type of ROS measurement used.
authors.affiliation: The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, Georgia, USA.
authors.forename: Ariel
authors.initials: A
authors.lastname: Kniss
authors.forename: Hang
authors.initials: H
authors.lastname: Lu
authors.forename: Dean P
authors.initials: DP
authors.lastname: Jones
authors.forename: Melissa L
authors.initials: ML
authors.lastname: Kemp
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D000968:Antimycin A / Q000494:pharmacology; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000032:analysis / Q000378:metabolism; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis* / Q000378:metabolism; D019169:Jurkat Cells; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008928:Mitochondria / Q000378:metabolism*; D061848:Optical Imaging / Q000295:instrumentation / Q000379:methods; D017382:Reactive Oxygen Species / Q000032:analysis* / Q000378:metabolism; D059010:Single-Cell Analysis / Q000295:instrumentation* / Q000379:methods; D013481:Superoxides / Q000032:analysis* / Q000378:metabolism; D014162:Transfection / Q000379:methods
chemical_list: D000900:Anti-Bacterial Agents; D005456:Fluorescent Dyes; D017382:Reactive Oxygen Species; D013481:Superoxides; D000968:Antimycin A; D006861:Hydrogen Peroxide