A Self-Sustaining Near-Infrared Afterglow Chemiluminophore for High-Contrast Activatable Imaging

Afterglow imaging holds great promise for ultrasensitive bioimaging due to its elimination of autofluorescence. Self-sustaining afterglow molecules (SAMs), which enable all-in-one photon sensitization, chemical defect formation and afterglow generation, possess a simplified, reproducible, and efficient superiority over commonly used multi-component systems. However, there is a lack of SAMs, particularly those with much brighter near-infrared (NIR) emission and structural flexibility for building high-contrast activatable imaging probes. To address these issues, this study for the first time reports a methylene blue derivative-based self-sustaining afterglow agent (SAN-M) with brighter NIR afterglow chemiluminescence peaking at 710 nm. By leveraging the structural flexibility and tunability, an activatable nanoprobe (SAN-MO) is customized for simultaneously activatable fluoro-photoacoustic and afterglow imaging of peroxynitrite (ONOO-), notably with a superior activation ratio of 4523 in the afterglow mode, which is at least an order of magnitude higher than other reported activatable afterglow systems. By virtue of the elimination of autofluorescence and ultrahigh activation contrast, SAN-MO enables early monitoring of the LPS-induced acute inflammatory response within 30 min upon LPS stimulation and precise image-guided resection of tiny metastatic tumors, which is unattainable for fluorescence imaging. mMB, as a novel self-sustaining afterglow molecule enabling photon sensitization, chemical defect formation, and afterglow generation, possessed a NIR afterglow chemiluminescence with a higher brightness and a NIR fluoro-photoacoustic signal. By introducing a responsive unit on the phenothiazine ring, it was tailored for ultrasensitive imaging of biomarkers with a superior activation ratio, enabling in vivo versatile imaging with high SBRs. image