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Evaluating H2O2 in Living Bacteria by Ratiometric Fluorescent Probe

Hydrogen peroxide (H2O2) is a significant signal molecule in physiological and pathological processes. Levels of H2O2 in bacteria are proved to be a key factor in immune response. To sum up, detection of H2O2 levels in living bacteria is remarkable for further study of its physiological and pathological effects. Herein, we propose a novel ratiometric fluorescent probe (Nahp) to detect H2O2 in living cells and bacteria. In addition, based on boronate, Nahp has satisfactory selectivity and sensitivity toward H2O2 (LOD = 0.158 μM). Furthermore, with excellent detection performance to H2O2, Nahp is successfully used for fluorescent bioimaging of H2O2 and measuring H2O2 accumulation in bacteria. Most importantly, the probe was also used to image H2O2 in three Gram-negative bacteria, clearly revealing for the first time significant differences in H2O2 expression levels in live bacteria.