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Merck
CN

Live-Cell Imaging of Mitochondrial Redox State in Yeast Cells.

STAR protocols (2020-12-31)
Pin-Chao Liao, Emily J Yang, Liza A Pon
ABSTRACT

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).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methyl viologen dichloride hydrate, 98%
Sigma-Aldrich
L-Tyrosine, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 99.0-101.0%
Sigma-Aldrich
L-Leucine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
D-(+)-Glucose, ≥99.5% (GC)
Sigma-Aldrich
DL-Dithiothreitol, for molecular biology, ≥98% (HPLC), ≥99% (titration)
Sigma-Aldrich
L-Methionine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
L-Lysine monohydrochloride, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
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L-Arginine, from non-animal source, meets EP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
Uracil, BioReagent, suitable for cell culture
Sigma-Aldrich
L-Tryptophan, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 99.0-101.0%
Sigma-Aldrich
L-Phenylalanine, from non-animal source, meets EP, JP, USP testing specifications, suitable for cell culture, 98.5-101.0%
Sigma-Aldrich
Adenine hemisulfate salt, ≥99%