Measurement of dynamic F-actin changes during exocytosis

Exocytosis requires the fusion of vesicle membrane to the cell membrane. It is tightly regulated and orchestrated in space and time by diverse cellular mechanisms. It has long been recognized that one of these mechanisms is an essential role played by the cytoskeleton. In particular, accumulating evidence shows that the F-actin network is engaged during the final stages of vesicle interactions with the cell membrane. Using a combination of methods it is now possible to gain insights into F-actin dynamics and reveal its role during exocytosis. Here, we describe the use of two-photon and confocal microscopy to visualize F-actin changes at the cell membrane during exocytosis.