Fluorescent measurement of [Ca²+]c: basic practical considerations

It is extremely difficult to write a prescriptive account of how to measure cytosolic-free Ca(2+) ([Ca(2+)](c)) that will suit all potential investigators. The problem arises because of the wide diversity of fluorescent Ca(2+) indicators that are now available, the variety of cells to be investigated, and the range of detection equipment that can be used. Consequently, this chapter is designed to provide the user with an overview of the technology in order that he or she can move toward developing a protocol that will suit the experimental objectives, the cells, and the equipment available to the investigator.