Compartmentalizing Genetically Encoded Calcium Sensors

Within single cells there is a complex myriad of signaling which controls physiological process many of which are modulated, or signaled directly, by intracellular calcium ions. Understanding the exquisitely sensitive, and spatially restricted, changes in calcium has been of interest to the researcher for a number of years. Recent advances in this field have been driven by the development of genetically encoded calcium probes for detecting calcium changes within the cells specifically targeting organelles such as mitochondria, endoplasmic reticulum, and the nucleus. In this chapter the authors outline some of the available fluorescent probes, with particular emphasis on an endoplasmic reticulum targeted calcium biosensor in cell signaling studies with astrocytes, detailing experimental protocols and the interpretation of data from such probes.