Description
Fluorescein diacetate (FDA) is a cell-permeant esterase substrate that can serve as a viability probe that measures both enzymatic activity, which is require to activate its fluorescence, and cell-membrane integrity, which is required for intracellular retention of their fluorescent product. Upon hydrolysis by intracellular esterases, this AM ester yields fluorescein.
Uses
Fluorescein diacetate is a non-fluorescent molecule, which is hydrolysed to fluorescent fluorescein in live cells. Dead cells cannot accumulate or hydrolyse FDA. It has been used for the differentiation of live cells from dead cells. It can be also used as a fluorogenic substrate for hGSTP1-1.
Biotechnological Applications
Fluorescein Diacetate Stain is an esterase substrate that can permeate cell membranes. In viable cells with an active metabolism, intracellular esterases hydrolyze Fluorescein Diacetate Stain to produce the green fluorescent compound fluorescein. Fluorescein accumulates in viable cells, causing them to fluoresce green.
Fluorescein Diacetate Stain can be used with Propidium Iodide Stain for Yeast to assess cell viability with the automated fluorescence cell counters of the LUNA® family. Viable cells convert fluorescein diacetate into fluorescein and thus fluoresce green. Nonviable cells take up propidium iodide and fluoresce red.
Fluorescence Signal Enhancer 1 can be used with Fluorescein Diacetate Stain to inhibit ATP-dependent fluorescein efflux from metabolically active cells. This will result in lower background and increased signal intensity, thereby enhancing the fluorescein signal-to-noise ratio.
