Construction of a super large Stokes shift near-infrared fluorescent probe for detection and imaging of superoxide anion in living cells, zebrafish and mice

As one of the major reactive oxygen species (ROS), superoxide anion (O-2(center dot-)) is engaged in maintaining redox homeostasis in the cell microenvironment. To identify the pathological roles in related disorders caused by abnormal expression of O-2(center dot-), it is of great significance to monitor and track the fluctuation of O-2(center dot-) concentration in vivo. However, the low concentration of O-2(center dot-) and the interference caused by tissue autofluorescence make the development of an ideal detection methodology full of challenges. Herein, a "Turn-On" chemical response near-infrared (NIR) fluorescence probe Dcm-Cu-OTf for O-2(center dot-) detection in inflamed models, was constructed by conjugating the NIR fluorophore (dicyanisophorone derivative) with an O-2(center dot-) sensing moiety (trifluoromethanesulfonate). Dcm-Cu-OTf exerted about 140-fold fluorescence enhancement after reacting 200 mu M O-2(center dot-) with an excellent limited of detection (LOD) as low as 149 nM. Additionally, Dcm-Cu-OTf exhibited a super large Stokes shift (260 nm) and high selectivity over other bio-analytes in stimulated conditions. Importantly, Dcm-Cu-OTf showed low toxicity and enabled imaging of the generation of O-2(center dot-) in the Lipopolysaccharide (LPS)-stimulated HeLa cells, zebrafish, and LPS-induced inflamed mice. The present study provided a potential and reliable detection tool to inspect the physiological and pathological progress of O-2(center dot-) in living biosystems.