Measurement of Calcium Fluctuations Within the Sarcoplasmic Reticulum of Cultured Smooth Muscle Cells Using FRET-based Confocal Imaging

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 cycling in a more direct fashion. For example, it's very important to know the relation with between our calcium transport in the sarcoplasmic reticulum and ER stress.

The advantage of this technique is that we can directly measure the calcium inside the lumen of the sarcoplasmic reticulum rather than to have to extrapolate it from indirect measurements of changes in the cytoplasm. Demonstrating this technique will be Gabriela Ziomek who is a graduate student in my laboratory. To begin this procedure, prepare a one to 25 dilution of protein mixture in DMEM.