Assembly-Improved Dye-Quenching Efficiency Enables High-Contrast Activatable Nanoreporters with Multimodal Readouts

Abstract Precise optical signal regulation is essential for diagnostic tools; however, most existing systems suffer from inefficient off-state quenching due to limited control over intermolecular interactions and are often constrained by single-modality outputs. Herein, we introduce a self-assembly guided quenching framework, termed Assembly-Improved Dye-quenching Efficiency (AIDE), which integrates Förster resonance energy transfer (FRET) and aggregation-caused quenching (ACQ) within a cooperative nanoscale architecture to enhance off-state suppression. As a representative implementation, AIDE is realized using tin(IV) naphthalocyanine (SnPc), a near-infrared chromophore with a high extinction coefficient and strong aggregation propensity. When combined with infrared fluorophores, SnPc functions as both a FRET acceptor and an ACQ scaffold, enabling near-complete fluorescence suppression (>99%) through assembly-dependent energy dissipation beyond conventional molecular designs. By introducing biomarker-cleavable linkers, we construct single-stimulus, dual-responsive multimodal nanoreporters (SDMRs) that integrate near-infrared fluorescence, photoacoustic imaging, and renal-clearable urinary readouts upon activation, thereby extending output signal wavelengths and expanding quencher functionality. Notably, SDMRs exhibit intrinsic tropism for inflamed tissues. In rodent models, this trimodal platform enables early detection of tumors (∼6 mm) and skin allograft rejection (24 h post-transplantation). Together, AIDE establishes a generalizable framework for high-contrast, multimodal, and minimally invasive disease diagnostics.