Preparation of 35 mm Glass-bottom Culture Dishes For FRET-based Confocal Microscopy
3:18
Transient Transfection With Adenoviral Vector Carrying D1SR Ca2+ Indicator
4:17
Measurement of SR Luminal Ca2+ Using FRET-based Confocal Imaging
6:53
Measurement of Cytoplasmic Ca2+
7:55
Results: Distribution of the SR Ca2+ Indicator D1SR in Rat Aortic Smooth Muscle Cells
9:23
Conclusion
Transcripción
The overall goal of this imaging technique is to monitor fluctuations in lumenal SR calcium levels in response to calcium mobilizing agents specifically in vascular smooth muscle cells. This method can be helpful in elucidating cellular calcium cy
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Currently, most available calcium indicators are used to quantify cytoplasmic calcium transients as indirect measures of calcium released from the sarcoplasmic reticulum in cultured smooth muscle cells. This protocol describes the use of a specific FRET-based indicator that allows direct measurement of calcium signals within the sarcoplasmic reticulum lumen.