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Mercury (Hg) is considered an extremely toxic heavy metal which is extremely harmful to both the human body and environment. In addition, Hg 2+ -induced oxidative stress also exerts a crucial role to play in pathophysiological mechanisms of mercury toxicity. Thus, efficient and specific fluorescent probes for imaging Hg 2+ -induced oxidative stress are necessary. In the present study, we rationally design a novel Hg 2+ -activated and ICT-based NIR emission fluorescent probe NIR-HO for sequentially monitoring the ONOO - level with a "dual-key-and-lock" strategy. The probe NIR-HO showed rapid response and excellent specificity and sensitivity for the detection of Hg 2+ and ONOO - in vitro. Cell imaging demonstrated that Hg 2+ -induced oxidative stress was involved in ONOO - upregulation. Also, GSH, NAC, and EDTA were employed as excellent detoxifying drugs against Hg 2+ -induced toxicity. Moreover, the probe NIR-HO was successfully used for imaging Hg 2+ and ONOO - in vivo. In brief, NIR-HO provides a simple and powerful approach which can be used to image Hg 2+ -induced oxidative stress in the pathological environment.