A Microfluidic Systems Biology Approach for Live Single-Cell Mitochondrial ROS Imaging

Methods Enzymol. 2013;526:219-30. doi: 10.1016/B978-0-12-405883-5.00013-2.

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.

MeSH terms

  • Anti-Bacterial Agents / pharmacology
  • Antimycin A / pharmacology
  • Equipment Design
  • Fluorescent Dyes / analysis
  • Fluorescent Dyes / metabolism
  • Humans
  • Hydrogen Peroxide / analysis*
  • Hydrogen Peroxide / metabolism
  • Jurkat Cells
  • Microfluidic Analytical Techniques / instrumentation*
  • Microfluidic Analytical Techniques / methods
  • Mitochondria / metabolism*
  • Optical Imaging / instrumentation
  • Optical Imaging / methods
  • Reactive Oxygen Species / analysis*
  • Reactive Oxygen Species / metabolism
  • Single-Cell Analysis / instrumentation*
  • Single-Cell Analysis / methods
  • Superoxides / analysis*
  • Superoxides / metabolism
  • Transfection / methods

Substances

  • Anti-Bacterial Agents
  • Fluorescent Dyes
  • Reactive Oxygen Species
  • Superoxides
  • Antimycin A
  • Hydrogen Peroxide