5(6)-CFDA 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, the 5(6)-carboxyfluorescein diacetate yields 5(6)-carboxyfluorescein. As compared with fluorescein, carboxyfluorescein contains extra negative charges and is therefore better retained in cells.
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