NIR‐II Orthogonal Fluorescent Ratiometric Nanoprobe for In Situ Bioimaging of Carbon Monoxide

Carbon monoxide (CO) functions as a significant endogenous cell signaling molecule and is strongly associated with many physiological and pathological processes. However, conventional fluorescence imaging in the visible and near-infrared (NIR) I regions suffers autofluorescence background and photon scattering, hindering the accurate detection of CO in vivo. In addition, the complexity of physiological environments leads to fluctuating fluorescence emission. To solve these problems, herein, the NIR-II fluorescent nanoprobe NP-Pd for in vivo ratiometric bioimaging of CO is developed. In the presence of CO, NP-Pd exhibits responsive enhancement in absorption at 808 nm, which amplifies the fluorescence signal of down-conversion nanoparticles (DCNP) at 1060 nm under 808 nm excitation, while the fluorescence signal of DCNP at 1525 nm under 980 nm excitation remains unchanged and serves as an internal standard. Through this orthogonally ratiometric fluorescence strategy, accurate CO bioimaging and precise diagnosis of acute liver injury diseases are achieved in the mouse model experiments, providing a novel tool for the in vivo detection of CO-related diseases.