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JoVE Journal Biology
Cellular Redox Profiling Using High-content Microscopy
 

Cellular Redox Profiling Using High-content Microscopy

Article DOI: 10.3791/55449-v • 11:37 min • May 14th, 2017
May 14th, 2017 •
1,2, 1,2, 1, 3, 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

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요약
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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).

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Keywords: Cellular Redox Profiling High-content Microscopy Mitochondrial Morphofunction Reactive Oxygen Species Redox Fingerprint Oxidative Stress Human Dermal Fibroblasts CM-H2DCFDA TMRM Multi-well Plate Microscope Setup Imaging Protocol Well Plate Alignment Sequential Wavelength Acquisition
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