Designed fluorescent indicators are the basis for a major new technique in cell physiology, the quantitative measurement and dynamic imaging of intracellular concentrations of important ions and messengers such as Ca2+, Na+, H+, and adenosine 3',5'-cyclic monophosphate. Molecular engineering has now produced indicators with quite good selectivity and sensitivity for these analytes. In many cases, these probes can be introduced into large populations of cells by means of membrane-permeant chemical derivatives, so that the plasma membrane need never be disrupted or physically breached at any point. Like many other optical microscopic techniques, fluorescent indicators are readily applied to study living cells and tissues, with an unparalleled combination of spatial and temporal resolution. They offer one of the few methods for continuous nondestructive monitoring of dynamic intracellular biochemistry and signal transduction in single cells or subregions of cells.

Tsien, R. Y. (1992). Intracellular signal transduction in four dimensions: from molecular design to physiology. American Journal of Physiology-Cell Physiology, 263(4), C723–C728.