Cascade‐Amplified Ratiometric Fluorescent Probe for Time‐Locked Imaging Epilepsy

Accurate localization of the epileptogenic network (EN) is essential for surgical intervention in drug‐resistant epilepsy, which affects over 15 million people globally. However, current approaches are limited by their low specificity, insufficient sensitivity, and challenges in capturing transient epileptiform discharges, which results in surgical success rates ranging from 30% to 70%. Here, we present a cascade‐amplified ratiometric imaging strategy employing an engineered fluorescent polymeric probe (CAR‐FM) that tandemly targets two temporally distinct pathological features within the epileptogenic network: transient acidosis fluctuations triggered by discharges and sustained neuroinflammation. This dual‐cascade amplification enables CAR‐FM to achieve more than a >150‐fold enhancement in fluorescence ratio with markedly improved specificity and sensitivity. In addition, CAR‐FM demonstrates robust applicability across four epilepsy models, which enables time‐locked EN delineation for up to 100 h. In epileptic mouse models, CAR‐FM‐guided resection yields a 67% greater reduction in seizure burden compared to that of conventional electrocorticography. Given its ultrahigh imaging sensitivity and biocompatibility, CAR‐FM offers a clinically translatable strategy for intraoperatively refining EN visualization, which potentially expands the population eligible for epilepsy surgery.