A novel near-infrared fluorescent probe (SWJT-2) has been designed and synthesized for the detection of methylglyoxal (MGO). It showed a low detection limit (0.32 μM), high selectivity and the fastest detection (15 min) over various reactive carbonyl compounds in aqueous solution. SWJT-2 had been successfully applied to bioimaging in HeLa cells to detect exogenous and endogenous MGO.