Hypochlorous acid (HClO) as a biomarker of inflammation has been implicated in redox signaling and combating microbial infection. Therefore, it is of great significance to develop an efficient method for detection and analysis of HClO in osteoarthritis. Herein, a new”turn-off” mitochondria-targetable NIR fluorescent probe, NIR-ClO, was reported for specific analysis and imaging of osteoarthritis response-related HOCl levels in vitro and in vivo. In the presence of HClO, due to the specific HOCl-triggered C = C bond cleavage reaction, NIR-ClO obtained a high sensitive and selective fluorescence “On-Off” response toward HClO with a good limit of detection(LOD) as low as 28.3 nM, and showed a fast response time (<60 s) , which allow it to be used for detection of HClO under a simulated physiological condition. In addition, NIR-ClO was successfully used to imaging of HClO in living RAW264.7 cells and osteoarthritis model rat. The results suggest that NIR-ClO is a robust tool for future studies on diagnosis osteoarthritis.