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Chapters
- 00:05Title
- 01:15Loading Cells with Calcium Indicators
- 02:23Microinjecting Cells
- 03:42Co-immunofluorescence Studies
- 04:49Fast Calcium Imaging and Data Analysis
- 06:28Dissecting the Timing of Ca++ Fluxes Facilitates Evaluation of Their Impact on Cellular Events
- 11:22Conclusion
We present a method to isolate rapid (microsecond) calcium events from slower fluxes in living cells using laser scanning confocal microscopy. The method measures fluorescence intensity fluctuations of calcium indicators by recording line scans of several hundred pixels in a cell. Histogram analysis allows us to isolate the time scales of different calcium fluxes.
Tags
Calcium FluxesCardiomyocytesMeasuringFastCa2+ ActivityPhospholipase Cβ- Gαq PathwayAcetylcholineCaveolaeEntrapmentGαqProlonged Ca2+ SignalsDynamic Calcium ResponsesFluorescent Ca2+ IndicatorCa2+ GreenFluorescence Emission IntensityLine-scan ModeLaser Scanning Confocal MicroscopeTime CourseMicrosecond Time ScaleExcel AnalysisSigmaplot AnalysisFlux Rate
