4’,6-diamidino-2-phenylindole (DAPI) is a blue fluorescent nucleic acid stain that binds to double stranded DNA and appears to associate with AT clusters in the minor groove of the DNA molecule. DAPI undergoes approximately 20-fold enhancement of fluorescence when associated with DNA, having an excitation maximum of 358 nm and an emission maximum of 461 nm. The spectral properties of DAPI make it very suitable for detecting viability on a flow cytometer equipped with an ultraviolet or a violet laser. DAPI is an impermeant DNA dye; it is largely excluded from live cells by competent plasma membranes, but can enter a compromised membrane where the dye can interact with DNA in the cell. Under recommended conditions, dead cells produce a bright fluorescent signal when excited with a violet laser and detected in the wavelength range of 425-475 nm. It can be readily used with dyes excited by other laser lines. Apoptotic, necrotic cells and / or damaged cells are source of interference in the analysis of viable cells by flow cytometry. Non-viable cells can be evaluated and discriminated following DAPI positive labeling when viable cells remain unstained (negative).
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