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Chapters
- 00:05Title
- 01:33Imaging Plate Preparation
- 02:29Imaging Parameters for the Zeiss LSM 710 Confocal Microscope
- 03:37Optimizing the Imaging Parameters
- 04:49Mitochondrial-fusion Assay Set-up
- 07:36Analyzing the PA-GFP Signal Intensity
- 08:17Results: Representative Mitochondrial-fusion Assays
- 09:49Conclusion
Mitochondrial fusion was measured by tracking the equilibration of photoconverted matrix-targeted GFP across the mitochondrial network over time. Thus far, only one cell could be subjected to an hour long kinetic analysis at a time. We present a method that simultaneously measures multiple cells, thereby speeding up the data collection process.
Tags
FasterHigh ResolutionMtPA-GFP-based Mitochondrial Fusion AssayKinetic DataMultiple CellsParallelConfocal MicroscopyFusion ActivityPhoto-conversionPhotoactivatable GFP (mtPAGFP)EquilibrationLiving CellsTime Lapse ApproachScale UpAutomateLow Concentration Of TMRESignal IntensityPAGFP Expressing CellsSubcellular QuantificationCytoarchitectural Environment
