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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
Chapters
- 00:05Title
- 00:47Seeding Cells in a 96-well Plate
- 01:48Setting Up the Microscope and Acquisition Protocol
- 03:10Loading of the Cells with CM-H2DCFDA and TMRM
- 04:18First and Second Live Imaging Rounds
- 06:22Image Processing and Analysis
- 09:35Results: Effect of Saquinavir on Primary Human Fibroblasts
- 10:38Conclusion
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).
