Live-Cell Imaging of Mitochondrial Redox State in Yeast Cells

STAR Protoc. 2020 Nov 9;1(3):100160. doi: 10.1016/j.xpro.2020.100160. eCollection 2020 Dec 18.


The redox state of mitochondria is one indicator of the functional state of the organelles. Mitochondria are also the primary endogenous source of reactive oxygen species (ROS). Therefore, the redox state of the organelles also reflects their function in ROS production. Here, we provide step-by-step protocols for live-cell imaging and quantification of mitochondrial redox state using the genetically encoded fluorescent biosensor, mitochondria-targeted redox sensing GFP (mito-roGFP), and mitochondrial ROS using the membrane-permeant small molecule dihydroethidium (DHE) in budding yeast cells. For complete details on the use and execution of this protocol, please refer to Liao et al. (2020c).

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Ethidium / analogs & derivatives
  • Ethidium / metabolism
  • Green Fluorescent Proteins / metabolism
  • Imaging, Three-Dimensional*
  • Microbial Viability*
  • Mitochondria / metabolism*
  • Oxidation-Reduction
  • Saccharomyces cerevisiae / growth & development
  • Saccharomyces cerevisiae / metabolism*
  • Superoxides / metabolism


  • dihydroethidium
  • Superoxides
  • Green Fluorescent Proteins
  • Ethidium