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Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor
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Biología
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JoVE Revista Biología
Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor

Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor

DOI:
10.3791/2579-v

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06:26 min

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April 01, 2011

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1Department of Neuroscience and Cell Biology,University of Texas Medical Branch

Capítulos

  • 00:05Título
  • 01:19Pericam-mt Transfection and Cell Preparation
  • 04:11Image Processing and Analysis
  • 04:37Fluorescence Imaging Results
  • 05:58Conclusion

Summary

Traducción Automática

Traducción Automática

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

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