Synergistic Probe Combining Lysosome Anchoring and Tumor Defense System Targeting for Precise Tumor Visualization.

Precise tumor imaging is essential for accurate intraoperative decision-making, thereby directly influencing patient prognosis. Optical molecular probes enabling non-invasive, dynamic assessment of cancerous lesions are clinically crucial. However, current optical molecular probes face challenges in dodging false-positive and false-negative signals at once during imaging, limiting their clinical diagnostic use. Here, we introduce a class of lysosome-targeted, activatable optical probes (Dx-NH2), leveraging the increased lysosomal abundance during tumor metabolic reprogramming. Lysosomal protonation retains probes for better single-cell resolution and fewer false-negative signals. To enable more high-precision tumor imaging, we synthesized probe CN-D-GGT from CN-D-NH2 by introducing a γ-glutamyltransferase substrate, following our group's prior offensive and defensive integration strategy. Due to its smart design, CN-D-GGT, upon activation, shows marked changes in fluorescence and photoacoustic signals, enabling its application for multi-modal imaging. It also has high specificity, distinguishing cancer cells in co-culture (∼6-fold) and clearly differentiating tumors from normal and inflamed tissues (T/N or T/I signal ratios > 3.5). Importantly, it can also differentiate cancerous from adjacent tissues in clinical samples. This work has developed a probe that can accurately light tumors in complex pathological environments, with the potential to assist in intraoperative resection decision-making, avoid excessive or missed resection.