DAF-FM diacetate (4-amino-5-methylamino-2',7'-difluorofluorescein diacetate) is a cell-permeable fluorescent probe for highly sensitive nitric oxide (NO) detection (limit ~3 nM). Upon entering cells, it is converted to a cell-impermeant, NO-reactive form (excitation/emission at 495/515 nm). DAF-FM offers superior photostability, 160-fold fluorescence increase upon NO binding, and pH-independence above 5.5 for reliable imaging.
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