Construction of a dual-excitation ratiometric fluorescent probe for determining peroxynitrite levels in living cells and zebrafish.

Peroxynitrite (ONOO-) is a highly reactive oxygen species that plays a critical role in many physiological and pathological processes of cell function. This study aimed to propose a ratiometric fluorescent probe BDHCA derived from coumarin for determining the ONOO- level. ONOO- could specifically induce oxidative cleavage of the conjugated C = C double bond in probe BDHCA, providing a fluorescent ratiometric output. The response of probe BDHCA to ONOO- was selective, fast, and highly sensitive, with a detection limit of 50.3 nM. Biological imaging experiments suggested that probe BDHCA could be used to image ONOO- in living RAW264.7 cells and zebrafish.