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Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy
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Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy

Simultaneous Measurement of Mitochondrial Calcium and Mitochondrial Membrane Potential in Live Cells by Fluorescent Microscopy

DOI:
10.3791/55166-v

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08:43 min

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January 24, 2017

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, ,
1Centre for Genetic Diseases,Hudson Institute of Medical Research, 2The Department of Molecular and Translational Sciences,Monash University, 3Department of Physiology,University College London

Chapters

  • 00:05Title
  • 00:54Preparation of Cells
  • 02:21Staining of Cells with TMRM and Fluo-4 AM
  • 03:46Cell Imaging and Analysis
  • 06:44Results: Measuremnt of Mitochondrial Membrane Potential and Calcium in Digitonin Permeabilized 143B Cells
  • 07:57Conclusion

Summary

Automatic Translation

Automatic Translation

Mitochondria can utilize the electrochemical potential across their inner membrane (ΔΨm) to sequester calcium (Ca2+), allowing them to shape cytosolic Ca2+ signaling within the cell. We describe a method for simultaneously measuring mitochondria Ca2+ uptake and ΔΨm in live cells using fluorescent dyes and confocal microscopy.

Tags

Mitochondrial CalciumMitochondrial Membrane PotentialFluorescent MicroscopyLive Cell ImagingCalcium RegulationMitochondrial DiseaseCalcium-free HBSSTMRMFluo 4 AMDigitoninThapsigargin

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