Simultaneously detecting two biomarkers can significantly enhance disease diagnosis accuracy and deepen our understanding of disease pathogenesis. Developing a single fluorescent probe with two independent response units for this purpose is crucial. In line with this, a novel fluorescent probe (TPA-DT-DNH) had been proposed, featuring triphenylamine and C=N groups sensitive to lipid droplets (LDs) and hypochlorous acid (HClO), respectively. Spectral results highlighted that only the co-presence of LDs and HClO could “turn-on” the fluorescence of TPA-DT-DNH, when the concentration of one substance remained constant, the fluorescence intensity linearly increased with the concentration of the other substance. The imaging results demonstrated that TPA-DT-DNH can differentiate between normal cells and cells rich in LDs when HClO was present, and can detect HClO in HeLa cells. Furthermore, dynamic changes in the inflammation process induced by LPS can be visualized in both cells and zebrafish. Additionally, significant fluorescence enhancement was observed in atheromatous plaque.