Bisulfite (HSO3−) has been widely used in food and industry, which has brought convenience to human life, but also seriously endangered human health. In this work, the probe PBI was designed and synthesized to detect bisulfite (HSO3−) through nucleophilic addition reaction. The probe PBI showed a selective reaction to HSO3− and can quantitatively detect HSO3−. At the same time, the color of the probe PBI changed significantly, which provided a simple method for the naked eye to identify HSO3−. Finally, it was successfully applied to the fluorescence imaging of HSO3− in living cells.