Ca2+-sensitive fluorescent dyes and intracellular Ca2+ imaging

Imaging Ca(2+)-sensitive fluorescent indicators provides a common approach for studying Ca(2+) signals in many contexts. Fluorescent indicators are particularly useful for measuring acute Ca(2+) changes in a relatively noninvasive manner. The availability of indicators that can be targeted to specific cellular domains, coupled with variations in affinity, brightness or spectral characteristics, provides tools for exploring spatially and temporally diverse Ca(2+) signals, and moreover, multiplexing the readout of Ca(2+) with other cellular functions. This article aims to give the novice experimentalist some insight into the considerations and potential pitfalls that impinge on the use of fluorescent Ca(2+) indicators.