Dual-Response Functionalized Mitochondrial Fluorescent Probe for a Double Whammy Monitoring of Hypochlorite and Sulfur Dioxide in Heat Shock via Time Scales

Heat shock seriously affects the normal functioning of an organism and can lead to damage and even death in severe cases. To prevent or treat heat shock-related diseases, we require a better understanding of the mechanism of thermocytotoxicity. Here, we designed a functionalized dual-response fluorescent probe (<b>HCy-SO₂-HClO</b>) that could individually or simultaneously detect hypochlorous acid (HClO) and sulfur dioxide (SO₂) without interfering with each other and achieved the simultaneous tracing of both during the heat shock process for the first time. The introduction of the sulfonate group greatly increased the water solubility of the probe. Furthermore, the probe could effectively accumulate in the mitochondrial region. <b>HCy-SO₂-HClO</b> could respond to HClO and SO₂ within 10 s and 20 min, respectively, realizing a double whammy detection of both on the time scale. <b>HCy-SO₂-HClO</b> exhibited high specificity and sensitivity for HClO and SO₂. The highly biocompatible probe <b>HCy-SO₂-HClO</b> successfully achieved the detection of endogenous and exogenous SO₂ and HClO in living cells and in zebrafish. Moreover, the simultaneous detection of HClO and SO₂ in heat shock cells and mouse intestines was realized for the first time. This probe has achieved the detection of dual markers, which enhanced the accuracy and precision of disease detection and could serve as an effective research tool to prevent heat stroke-related diseases.