A Near-Infrared (NIR) ratiometric and on-off fluorescent probe 1 was synthesized, which exhibited high sensitivity and excellent selectivity in arginine (Arg) recognition. 1 can quantitatively detect Arg in concentration range from 0 to 150 μM with the LOD of 138 nM. Further, it displayed excellent selectivity and antiinterference over many biological species, metal ions and anions. Due to its ability to target lysosome, the response of Arg to this probe in a living cell was successfully tracked in a ratiometric manner via fluorescence imaging.