Hoechst 33258 is a blue-emitting fluorescent dye that selectively binds to adenine-thymine-rich regions in double-stranded DNA. Its fluorescence is enhanced by AT-rich DNA strands. With excitation/emission maxima at 351/463 nm, it exhibits a considerable Stokes shift. Although it can enter living cells, its permeability is lower than Hoechst 33342. Notably, Hoechst 33258 is less toxic than DAPI, ensuring better cell viability.
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