Induction of Ca-Dependent Exocytotic Processes by Laser Ablation of Endothelial Cells.

Ca regulates a variety of cellular processes that are essential to maintain cell integrity and function. Different methods have been used to study these processes by increasing intracellular Ca levels. Here, we describe a protocol to initiate Ca-dependent membrane-related events, using laser ablation by near-infrared irradiation. This creates a rupture in the plasma membrane that allows the extracellular Ca to enter the cell and thereby induce a receptor-independent Ca increase. We report laser ablation protocols to study two different Ca-induced processes in human endothelial cells-membrane resealing and exocytosis of secretory granules called Weibel-Palade bodies (WPBs). Thus, laser ablation represents a technique that permits the analysis of different Ca-regulated processes at high spatiotemporal resolution in a controlled manner.