Hypochlorous acid (HClO) is an important reactive oxygen species (ROS) and plays a significant role in living organisms. Highly selective and lysosome-targetable probes for sensing hypochlorous acid are rare. In this article, we designed and prepared a new lysosome-targeting and ratiometric fluorescent probe for monitoring the levels of hypochlorous acid. 4-Aminonaphthalimide was chosen as the fluorescent group and (2-aminoethyl) thiourea group was used as a specific recognition group for HClO. A morpholine unit was employed as a lysosome-targeting group. In the absence of HClO the probe underwent intramolecular charge transfer (ICT) and showed a green emission. When excess HClO is present, the ICT process was interrupted which caused a 57 nm blue-shift of fluorescence emission from 533 nm to 476 nm. The ratiometric fluorescent probe showed outstanding selectivity toward HClO over other various bioactive species. Furthermore, the ratiometric fluorescent probe exhibited rapid response time and ability of working in a wide pH range. The linear response of I476nm/I533nm toward HClO was obtained in a concentration range of HClO from 1.0 × 10-6 to 1.0 × 10-4 mol·L-1. The detection limit was estimated to be 8.0 × 10-7 mol·L-1 for HClO. Moreover, the probe showed a perfect lysosome-targeting ability, and has been successfully used to the confocal imaging of HClO in lysosomes of HepG2 cells with little cell toxicity. All of such good properties illustrated that it could be applied to determine HClO at lysosomes in living cells.