| Summary: | Inflammation is a vital protective response in living systems and closely related to various diseases. As a member of the reactive oxygen species (ROS) family, peroxynitrite (ONOO<sup>−</sup>) is involved in the organism’s inflammatory process and considered as an important biomarker of inflammation. Therefore, the construction of a simple, rapid, and sensitive tool for detecting ONOO<sup>−</sup> is of great importance for the diagnosis of inflammation. In this study, we constructed the new near-infrared fluorescence probe <b>BDP-ENE-S-Py<sup>+</sup></b> based on BODIPY dye, which has the advantages of fast response speed (2 min), good selectivity, and a high signal-to-noise ratio. Moreover, the probe had a good linear relationship (LOD = 120 nM) when the ONOO<sup>−</sup> concentration was 10–35 µM. In addition, <b>BDP-ENE-S-Py<sup>+</sup></b> could detect exogenous ONOO<sup>−</sup> in liver cancer cells without interference from other reactive oxygen species and visualize the fluctuations in ONOO<sup>−</sup> concentrations in cells. More importantly, <b>BDP-ENE-S-Py<sup>+</sup></b> was able to track the upregulation of ONOO<sup>−</sup> content in a mouse model of peritonitis induced by LPS. This work demonstrated that the near-infrared fluorescent probe for visualizing ONOO<sup>−</sup> level fluctuations could provide a promising tool for inflammation-related studies.
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