Real-time imaging of intracellular cysteine level fluctuations during Cu²⁺ or H₂O₂ induced redox imbalance using a...

Redox balance is a necessary guarantee to maintain the normal physiological activities of organisms. Cysteine (Cys), a critical biological thiol, has the effect of maintaining redox balance in the body. The concentration of intracellular Cys is abnormal under redox imbalance, thereby resulting in multiple diseases. Additionally, studies have revealed that Cu²⁺ can stimulate the body to produce excess reactive oxygen species (ROS, similar to H₂O₂), and the generated ROS will consume reducing substances (such as Cys) in the body, leading to redox imbalance. Thus, finding a simple and effective method to monitor Cys under redox imbalance is pressing. Here, a turn on probe (DDNO) was proposed by connecting SBD-Cl to a red dye (HDM). The probe can specifically recognize Cys with rapid response (180 s) and low detection limit (0.61 μM) through substitution-rearrangement reaction between sulfhydryl and chlorine atom. Bioimaging experiments indicated that the probe has good biocompatibility and cell membrane permeability, which can be applied to monitor the fluctuation of Cys levels in live cells and zebrafish under the redox imbalance induced by Cu²⁺ or H₂O₂.