Cellular Redox Profiling Using High-content Microscopy

Tom Sieprath; Tobias Corne; Joke Robijns; Werner J. H. Koopman; Winnok H. De Vos

doi:10.3791/55449

Journal

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Biology

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Cellular Redox Profiling Using High-content Microscopy

JoVE Journal
Biology

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JoVE Journal Biology

Cellular Redox Profiling Using High-content Microscopy

10,906 Views

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11:37 min

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May 14, 2017

DOI:

10.3791/55449-v

DOI:

10.3791/55449-v

•

11:37 min

May 14, 2017

•

10896 Views

Tom Sieprath^1,2, Tobias Corne^1,2, Joke Robijns¹, Werner J. H. Koopman³, Winnok H. De Vos^1,2

¹Laboratory of Cell Biology and Histology, Department of Veterinary Sciences,University of Antwerp ²Cell Systems and Imaging Research Group (CSI), Department of Molecular Biotechnology,Ghent University ³Department of Biochemistry , Radboud Institute for Molecular Life Sciences,Radboud University Medical Center

Summary

This paper presents a high-content microscopy workflow for simultaneous quantification of intracellular ROS levels, as well as mitochondrial membrane potential and morphology – jointly referred to as mitochondrial morphofunction – in living adherent cells using the cell-permeant fluorescent reporter molecules 5-(and-6)-chloromethyl-2',7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-H₂DCFDA) and tetramethylrhodamine methylester (TMRM).