Hydrogen peroxide (H₂O₂) plays a significant role in regulating a variety of biological processes. Dysregulation of H₂O₂ can lead to various diseases. Although numerous fluorescent imaging probes for H₂O₂ have been reported, the development of H₂O₂ ratiometric fluorescent probe with large Stokes shift remains rather limited. Such probes have shown distinct advantages, such as minimized interference from environment and improved signal-to noise ratio. In this work, we reported a new pyrene-based compound Py-VPB as H₂O₂ fluorescent probe in vitro. The probe demonstrated ratiometric detection behavior, large Stokes shift and large emission shift. In addition, the probe showed high sensitivity and selectivity towards H₂O₂ in vitro. Based on these excellent properties, we successfully applied Py-VPB to the visualization of exogenous and endogenous H₂O₂ in living cells. Cell imaging study also showed that our probe was localized in the mitochondria. We envision that the probe can provide a useful tool for unmasking the biological roles of mitochondrial H₂O₂ in living systems.

过氧化氢（H ₂ O ₂）在调节各种生物过程中起着重要作用。H ₂ O _2的失调可导致多种疾病。尽管已经报道了许多用于H ₂ O _2的荧光成像探针，但是具有大斯托克斯位移的H ₂ O ₂比例荧光探针的发展仍然相当有限。这样的探头显示出明显的优势，例如最小化来自环境的干扰和改善的信噪比。在这项工作中，我们报道了一种新的pyr基化合物Py-VPB作为H ₂ O ₂体外荧光探针。该探针显示出比率检测行为，大斯托克斯位移和大发射位移。另外，该探针在体外对H ₂ O ₂表现出高灵敏度和选择性。基于这些优异的性能，我们成功地将Py-VPB应用于可视化活细胞中外源性和内源性H ₂ O ₂。细胞成像研究还表明，我们的探针位于线粒体中。我们设想该探针可以为揭示线粒体H ₂ O ₂在生命系统中的生物学作用提供有用的工具。