Molecular Crystal Engineering of Organic Chromophores for NIR-II Fluorescence Quantification of Cerebrovascular Function

Although molecular design strategies for highly bright near-infrared II (NIR-II) fluorophores were proposed, the lack of solid structural identification (single crystal) hinders the further development of this field. This thorny issue is addressed by performing the structure-function relationship of NIR-II dyes, as confirmed by molecular single crystal engineering. Single crystal structure analysis confirms that twisted architectures (large dihedral angles ∼70°) and loose packing patterns (intermolecular distance of ∼3.4-4.5 Å) are key elements to enhance the absolute quantum yield (QY) in the solid state. Through regulating donor-acceptor distance and donor-acceptor interactions, the resultant well-defined TBP-<i>b</i>-DFA fluorophore displays an absolute QY of 0.4% with an emission extending to 1400 nm, which is favorable for NIR-II bioimaging. The cerebrovascular function, including cerebral blood flow and cerebrovascular reactivity of different conditions, is accurately quantified by a NIR-II fluorescence wide-field microscope. Our study provides a sight for designing NIR-II fluorophores, which is useful for studying cerebrovascular function.