A subscription to JoVE is required to view this content. Sign in or start your free trial.
Chapters
- 00:05Title
- 01:08Preparation of 35 mm Glass-bottom Culture Dishes For FRET-based Confocal Microscopy
- 03:18Transient Transfection With Adenoviral Vector Carrying D1SR Ca2+ Indicator
- 04:17Measurement of SR Luminal Ca2+ Using FRET-based Confocal Imaging
- 06:53Measurement of Cytoplasmic Ca2+
- 07:55Results: Distribution of the SR Ca2+ Indicator D1SR in Rat Aortic Smooth Muscle Cells
- 09:23Conclusion
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.
