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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
Dieses Papier präsentiert einen hochauflösenden Mikroskopie-Workflow zur gleichzeitigen Quantifizierung von intrazellulären ROS-Niveaus sowie mitochondrialem Membranpotential und Morphologie - gemeinsam als mitochondriale Morphofunktion bezeichnet - in lebenden adhärenten Zellen unter Verwendung der zellpermeablen fluoreszierenden Reportermoleküle 5- (und- 6) -chlormethyl-2 ', 7'-dichlordihydrofluoresceindiacetat, Acetylester (CM-H 2 DCFDA) und Tetramethylrhodaminmethylester (TMRM).
