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Despite the wide range of applications of bright NIR‐II polymethine scaffolds in biomedical imaging, their solvatochromism and aggregation‐caused quenching (ACQ) effects in aqueous solutions limit their inherent brightness using traditional encapsulation methods, and effective hydrophilization strategies are still scarce. Here, a new set of Flav dyes is synthesized and PEGylated, followed by manufacturing DSPE@FlavP2000 nanoparticles using a self‐adaptive co‐assembly strategy to overcome these limitations. FlavP2000 can autonomously adjust its conformation when co‐assembled with DSPE‐PEG2000, resulting in high‐efficiency luminescence (≈44.9% fluorescence of Flav in DMSO). DSPE@FlavP2000 enables NIR‐IIb (>1500 nm) angiography with high signal‐to‐noise ratios. Notably, this co‐assembly can occur in situ between FlavP2000 with proteins in the living body based on a novel mechanism of brightness activation induced by disassembly (BAD), achieving consistent brightness as DSPE@FlavP2000 in blood or serum. The self‐adaptive co‐assembly strategy can be enhanced by incorporating an IPA moiety, which dynamically binds to albumin to prolong the dye's blood circulation time. Thus, the “enhanced” BAD is successfully applied to long‐term vascular imaging and sciatic nerve imaging. Both the self‐adaptive co‐assembly strategy and BAD phenomenon improve the selectivity and availability of the hydrophilization methods, paving the way for efficient biological applications of polymethine dyes.
Using a self‐adaptive co‐assembly strategy, FlavP2000 can autonomously adjust its conformation when co‐assembled with DSPE‐PEG2000, resulting in high‐efficiency luminescence which enables NIR‐IIb (>1500 nm) angiography with ultra‐high signal‐to‐noise ratios. Notably, this co‐assembly can also occur in situ between IPA‐FlavP2000/FlavP2000 with proteins in the living body based on a novel mechanism of brightness activation induced by disassembly (BAD).