A tri-channel ratiometric fluorescent probe with dual recognition sites for imaging exogenous and endogenous...

Hypochlorite is crucial in physiological and pathological processes, necessitating tools for its detection in living cells. We developed PZ-DPK, a novel ratiometric fluorescent probe, for selective and sensitive detection of exogenous and endogenous ClO^-. PZ-DPK demonstrated excellent biocompatibility, with cell viability exceeding 80% at 20 μM. Fluorescence imaging revealed that PZ-DPK penetrated cell membranes and reacted with ClO^-, producing distinct signals in red, green, and blue channels. Exogenous ClO^- detection in HepG2 cells showed strong red channel signals, while green and blue channels indicated reaction products. For endogenous ClO^-, PMA-stimulated cells exhibited enhanced green and blue signals, whereas NAC pretreatment preserved red channel fluorescence. PZ-DPK's tri-channel imaging capability provides a powerful tool for studying ClO^- related biological processes.