Imbalanced iron homeostasis plays a crucial role in neurological diseases, yet direct imaging evidence revealing the distribution of active ferrous iron (Fe²⁺) in the living brain remains scarce. Here, we present a near-infrared excited two-photon fluorescent probe (FeP) for imaging changes of Fe²⁺ flux in the living epileptic mouse brain. In vivo 3D two-photon brain imaging with FeP directly revealed abnormal elevation of Fe²⁺ in the epileptic mouse brain. Moreover, we found that dihydroartemisinin (DHA), a lead compound discovered through probe-based high-throughput screening, plays a critical role in modulating iron homeostasis. In addition, we revealed that DHA might exert its antiepileptic effects by modulating iron homeostasis in the brain and finally inhibiting ferroptosis. This work provides a reliable chemical tool for assessing the status of ferrous iron in the living epileptic mouse brain and may aid the rapid discovery of antiepileptic drug candidates.

不平衡的铁稳态在神经系统疾病中起着至关重要的作用，但揭示活体大脑中活性亚铁 (Fe ^{2+ ) 分布的直接影像学证据仍然很少。}在这里，我们提出了一种近红外激发双光子荧光探针 (FeP)，用于成像活癫痫小鼠大脑中 Fe ^{2+通量的变化。}FeP 的体内3D 双光子脑成像直接显示 Fe ^{2+异常升高}在癫痫小鼠的大脑中。此外，我们发现双氢青蒿素 (DHA) 是一种通过基于探针的高通量筛选发现的先导化合物，在调节铁稳态中起关键作用。此外，我们发现 DHA 可能通过调节大脑中的铁稳态并最终抑制铁死亡来发挥其抗癫痫作用。这项工作为评估活癫痫小鼠大脑中亚铁的状态提供了一种可靠的化学工具，并可能有助于快速发现抗癫痫药物候选物。