Imaging calcium sparks in cardiac myocytes
- Methods Mol Biol. 2011:689:205-14. doi: 10.1007/978-1-60761-950-5_12.
- 1. Department of Physiology and Biophysics, ICB, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil. [email protected]
Calcium ions play fundamental roles in many cellular processes in virtually all type of cells. The use of CA(2+) sensitive fluorescent Indicators has proven to be an indispensable tool for studying the spatio-temporal dynamics of intracellular calcium ([CA(2+)](i)). With the aid of laser scanning confocal microscopy and new generation of CA(2+) Indicators, highly localized, short-lived CA(2+) signals, namely CA(2+) sparks, were revealed as elementary CA(2+) release events during excitation-contraction coupling in cardiomyocytes. Since the discovery of CA(2+) sparks in 1993, the demonstration of dynamic CA(2+) micro-domains in living cardiomyocytes has revolutionized our understanding of CA(2+)-mediated signal transduction in normal and diseased Hearts. In this chapter, we have described a commonly used method for recording local and global CA(2+) signals in cardiomyocytes using the fluorescent indicator fluo-4 acetoxymethyl (AM) and laser scanning confocal microscopy.