Hypochlorous acid (HClO) plays a critical role in physiological activities of maintaining the stable oxidation balance of organisms, which was proved to relate to some serious diseases. In this work, 4-nitrobenzenesulfonylhydrazide based fast-responsive two-photon fluorescent probe CoPh-ClO was designed and synthesized reasonably, which possessed low cytotoxicity, good anti-interference characteristics, a large Stokes shift (85 nm), and good two-photon performance. In addition, probe CoPh-ClO was successfully applied to detect exogenous HClO in living HeLa cells and endogenous HClO in living RAW264.7 cells respectively. Moreover, we successfully achieved tissues imaging with a deep penetration depth of 65 µm and zebrafish imaging accompanied with a high contrast (about 45-fold). Interestingly, the introduce of benzene ring between fluorophore and reaction site made probe CoPh-ClO more sensitive (only 20 s) with a large turn-on signal. The probe CoPh-ClO was modified and possessed better stability (more than 10 mins) even in excessive HClO. All of mentioned above merits demonstrated that CoPh-ClO could be a promising imaging tool for monitoring HClO in various physiological processes, and the introduction of benzene ring would provide a new perspective for the development of multi-function probes.

次氯酸（HClO）在维持生物体稳定氧化平衡的生理活动中起着至关重要的作用，已被证明与一些严重的疾病有关。本工作合理设计合成了基于4-硝基苯磺酰肼的快速响应双光子荧光探针CoPh-ClO，该探针具有低细胞毒性、良好的抗干扰特性、较大的斯托克斯位移（85 nm）和良好的双光子荧光探针。表现。此外，探针CoPh-ClO分别成功应用于检测活HeLa细胞中的外源性HClO和活的RAW264.7细胞中的内源性HClO。此外，我们成功地实现了深度穿透深度为 65 µm 的组织成像和伴随高对比度（约 45 倍）的斑马鱼成像。有趣的是，在荧光团和反应位点之间引入苯环使探针CoPh-ClO更加灵敏（仅 20 s），并具有较大的开启信号。探针CoPh-ClO进行了修改，即使在过量的 HClO 中也具有更好的稳定性（超过 10 分钟）。上述所有优点表明，CoPh-ClO可能成为监测各种生理过程中HClO的有前途的成像工具，苯环的引入将为多功能探针的开发提供新的视角。