Imaging Calcium Sparks in Cardiac Myocytes

Abstract

Calcium ions play fundamental roles in many cellular processes in virtually all type of cells. The use of Ca2+ sensitive fluorescent indicators has proven to be an indispensable tool for studying the spatio-temporal dynamics of intracellular calcium ([Ca2+]i). With the aid of laser scanning confocal microscopy and new generation of Ca2+ indicators, highly localized, short-lived Ca2+ signals, namely Ca2+ sparks, were revealed as elementary Ca2+ release events during excitation–contraction coupling in cardiomyocytes. Since the discovery of Ca2+ sparks in 1993, the demonstration of dynamic Ca2+ micro-domains in living cardiomyocytes has revolutionized our understanding of Ca2+-mediated signal transduction in normal and diseased hearts. In this chapter, we have described a commonly used method for recording local and global Ca2+ signals in cardiomyocytes using the fluorescent indicator fluo-4 acetoxymethyl (AM) and laser scanning confocal microscopy.