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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
Detta papper presenterar ett högt innehållsmikroskopi-arbetsflöde för samtidig kvantifiering av intracellulära ROS-nivåer, såväl som mitokondriell membranpotential och morfologi - gemensamt betecknad som mitokondriell morfofunktion - i levande vidhäftande celler med användning av cell-permeant fluorescerande reportermolekyler 5- (och- 6) -klorometyl-2 ', 7'-diklorodihydrofluoresceindiacetat, acetylester (CM-H2 DCFDA) och tetrametylrhodamin-metylester (TMRM).
