This homogeneous assay involves simply adding a single reagent, the CellCount-Blue reagent, to the cell culture and measuring the fluorescence intensity (excitation wavelength = 530 - 570 nm, emission wavelength = 590 - 620 nm) after an incubation step. The CellCount- Blue reagent, like other resazurin-based assays such as the Alamar Blue reagent, utilizes the redox dye resazurin which is not fluorescent, but upon reduction by metabolically active cells is converted into a highly fluorescent product (resorufin). Living cells can readily reduce this non-toxic reagent and the resulting increase in fluorescence intensity can be conveniently monitored using a fluorescence spectrophotometer or plate reader. Nonviable cells have no metabolic capacity and, thus, will not reduce the dye. Therefore, the fluorescence intensity observed in this assay is a true measure of the viable cells.
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