Live Cell Assay

The Live Cell (LC) Assay is a fluorescence-based method designed to quantify relative changes in live mammalian cell number following treatment with test compounds. This assay enables the assessment of compound-induced mitogenic or cytotoxic effects on cells. The protocol is optimized for use with adherent cells cultured in black or white 96-well plates. It provides a convenient and reliable tool for evaluating cell viability as part of screening workflows, mechanistic studies, or toxicity assessments.

The LC Assay utilizes the fluorogenic substrate Calcein-AM (AcetoxyMethyl ester of Calcein) to provide a sensitive, quantitative measure of the relative number of live Cells remaining in the assay wells following their exposure to test compounds. Calcein-AM is a hydrophobic, non-fluorescent molecule that readily crosses cellular membranes. Once in the intracellular environment, Calcein-AM is hydrolyzed by endogenous esterases in a time- and temperature-dependent manner. The resulting product, Calcein, is a hydrophilic fluorescent molecule. Due to the high charge density of the resulting reaction product, there will be no appreciable loss/efflux of Calcein from the intracellular compartment during the short reaction period of the LC Assay.