Multi-parametric NIR-II fluorescence perfusion imaging towards quantitative stroke evaluation

Preclinical stroke research is severely hampered by the lack of high-throughput, quantitative perfusion imaging tools capable of bridging the gap between functional hemodynamics and structural injury. The emerging second near-infrared (NIR-II) fluorescence imaging offers superior spatial resolution and tissue penetration, but its utility remains restricted by a lack of standardized analysis protocols and inability to resolve in-depth infarct regions. Here, we introduce FIMPA, an integrated analytical framework for standardized, multi-parametric stroke evaluation based on NIR-II fluorescence perfusion imaging. FIMPA systematically processes the image sequence through motion correction and atlas registration to generate an anatomically aligned library of 125 vascular and hemodynamic parameters. Statistical screening identifies 67 stroke-correlated features, enabling objective, data-driven quantification of perfusion deficits. To resolve spatial infarct boundaries, we developed FIMPA-SI (FIMPA Stroke Index), a deep learning-based index utilizing a ViT-UNETR framework. By leveraging self-supervised pre-training with MRI data, FIMPA-SI suppresses dominant vascular signals to translate multi-parametric dynamics into anatomically precise stroke severity maps. Validated in tMCAO mice, FIMPA-SI achieves high spatial concordance with MRI and significantly outperforms conventional laser speckle contrast imaging and single-parameter metrics. FIMPA provides a robust solution for accelerating preclinical therapeutics and advancing optical neuroimaging.