A fluorescent probe for monitoring Cys fluctuations in the oxidative stress environment simulated by Cu²⁺ or H₂O₂

Redox balance is the core of holding the good physiological state of the body. Cysteine (Cys) is one of the important biomolecules, which plays an indispensable role in maintaining the body's redox homeostasis. The redox of organisms is mainly the result of the dynamic balance between reactive oxygen species (ROS) and biological reducing agents (such as Cys). Fluorescent probes have the advantages of simple operation, good specificity and high sensitivity, and have become a common tool for bio-sensing in complex systems. In this article, we designed a probe NF-O-SBD that can specifically detect Cys. The chlorine atom of NF-O-SBD was easily substituted by sulfhydryl as a reaction site. After the formation of sulfur substitution products, intramolecular rearrangement occurred and fluorescent signal was emitted in the yellow channel at 550 nm. It can be seen from the spectroscopy experiment that the content of Hcy in organisms (15 μM) basically did not cause significant fluorescence changes, Therefore, based on the practical application in biology, we further used NF-O-SBD to visualize endogenous and exogenous Cys in HepG-2 cells and zebrafish. Simultaneously, we used Cu²⁺ or H₂O₂ induction to simulate the oxidative stress environment of cells and zebrafish, under which the concentration variation of Cys was monitored.