NEM assisted real-time fluorescence detection of Cys in cytoplasm and mice imaging by a Coumarin probe containing carboxyl group.

Alterations of the homeostasis balance of cysteine (Cys) are associated with a variety of diseases and cellular functions, and therefore, Cys dynamic real-time living cell intracellular imaging and quantification are important for understanding the pathophysiological processes. Thus, Cys probe that can permeate high efficiently is the first one to be affected. In fact, it is difficult for organic molecular probes to infiltrate cells because of the unique structure of the cell membrane. In this work, we found that probe containing-carboxyl just stagnated in cytomembrane due to carboxyl of probe and amino group of membrane protein forming peptide chains, nevertheless, the addition of NEM, improved membrane permeability by NEM reacting with sulfhydryl of membrane protein, which made probe permeate high efficiently and sequentially real-time detect the Cys in cytoplasm. It is the first time noted that NEM can regulate Cys probe containing-carboxyl for high efficient detection in cytoplasm. Additionally, probe was successfully applied to image Cys in mouse.