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Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor
Kapitel
- 00:05Titel
- 01:19Pericam-mt Transfection and Cell Preparation
- 04:11Image Processing and Analysis
- 04:37Fluorescence Imaging Results
- 05:58Conclusion
This protocol describes a method for real-time measurement of mitochondrial calcium fluxes by fluorescent imaging. The method takes advantage of a circularly permutated YFP-based dual-excitation ratiometric calcium sensor (ratiometric pericam-mt) selectively expressed in mitochondria.
Tags
MonitoringDynamic ChangesMitochondrial Calcium LevelsApoptosisGenetically Encoded Calcium SensorIntracellular Calcium ConcentrationStimuliCellular ProcessesProliferationDifferentiationCalcium AccumulationPro-apoptotic FactorsCytosolLiving CellsIn SituHigh-resolution Fluorescent ImagingDual-excitation RatiometricNon-ratiometric Synthetic Calcium Indicator DyesIntracellular Calcium SignalingCytosolic Calcium FluxesRhod-2Rhod-FFSynthetic IndicatorsMitochondriaRepetitive IncreasesMitochondrial MorphologyLeak Out Of MitochondriaLong-term ExperimentsGenetically Encoded Calcium IndicatorsGreen Fluorescent Protein (GFP)Aequorin
