A pyrene-based ratiometric fluorescent probe with a large Stokes shift for selective detection of hydrogen peroxide in living cells

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.