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

Abstract

As one of the major reactive oxygen species (ROS), superoxide anion (O₂^•-) is engaged in maintaining redox homeostasis in the cell microenvironment. To identify the pathological roles in related disorders caused by abnormal expression of O₂^•-, it is of great significance to monitor and track the fluctuation of O₂^•- concentration in vivo. However, the low concentration of O₂^•- 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 O₂^•- detection in inflamed models, was constructed by conjugating the NIR fluorophore (dicyanisophorone derivative) with an O₂^•- sensing moiety (trifluoromethanesulfonate). Dcm-Cu-OTf exerted about 140-fold fluorescence enhancement after reacting 200 μM O₂^•- 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 O₂^•- 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 O₂^•- in living biosystems.

Keywords: Acute inflammation; Fluorescence imaging; Living cells; NIR fluorescent probe; Superoxide anion.