Ag+ pollution is of great harm to the human body and environmental biology. Therefore, there is an urgent need to develop inexpensive and accurate detection methods. Herein, lignin-derived structural memory carbon nanodots (CSM-dots) with outstanding fluorescence properties were fabricated via a green method. The mild preparation process allowed the CSM-dots to remain plentiful phenol, hydroxyl, and methoxy groups, which have a specific interaction with Ag+ through the reduction of silver ions. Further, the sulfur atoms doped on CSM-dots provided more active sites on their surface and the strong interaction with Ag nanoparticles. The CSM-dots can specifically bind Ag+, accompanied by a remarkable fluorescence quenching response. This “turn-off” fluorescence behavior was used for Ag+ determination in a linear range of 5–290 μM with the detection limit as low as 500 nM. Furthermore, findings showed that this sensing nano-platform was successfully used for Ag+ determination in real samples and intracellular imaging, showing great potential in biological and environmental monitoring applications.