An abnormal endogenous SO2 level can induce toxicological effects leading to cancer, cardiovascular diseases, and neurological disorders, as they involve in various physiology and pathological processes in mitochondria cell apparatus. In this study, we report a ratiometric fluorescent probe (NAB-Id) based on the cyanine dye for the detection of sulfur dioxide (SO2) derivatives in aqueous media and live cells. Upon the addition of SO2 derivatives (SO32−/HSO3−) to the solution of NAB-Id, led to the nucleophilic addition to the polymethine, resulting in dramatic colorimetric and fluorescence dual responses. NAB-Id responses to HSO3− with low detection limit (58.6nM), outstanding specificity (without the interfering of H2S), fast response (t1/2 = 30s) and working well at physiological pH. Fluorescence co-localization studies demonstrated that NAB-Id was a mitochondria-targeted fluorescent probe for the real-time sensing and bioimaging of SO2 derivatives in living cells.