Nitric oxide (NO) participates in various pathways and revealing its dynamics is critical for resolving its pathophysiology. While there are methods available for detecting biological NO, few are capable of tracking NO dynamics. Herein, inspired by the cellular machinery of reversible thiol modification by NO, we have successfully designed a family of cysteine analogues tagged with fluorophores for visualizing cellular NO dynamics. Biocompatible probes 1a and 2a were found to be most tolerated by the cellular machinery to swiftly switch between their original fluorescent state and quenched S-nitrosated state in response to cellular NO dynamics. We showcased their applicability in various cell types and utility to track NO fluctuations in activated murine macrophages in response to an anti-inflammatory agent treatment. We expect that probes 1a and 2a will afford promising tools for studying NO pathophysiology by tracking its dynamics in health and disease.

一氧化氮（NO）参与各种途径，揭示其动力学对于解决其病理生理学至关重要。尽管存在用于检测生物NO的方法，但很少能够跟踪NO动态。在此，受NO可逆硫醇修饰的细胞机制的启发，我们已经成功设计了标记有荧光团的半胱氨酸类似物家族，用于可视化细胞NO动力学。生物相容性探针 1a 和 2a发现细胞对细胞NO动力学的响应最能被细胞机制耐受，以在其原始荧光状态和淬灭的S-亚硝化状态之间快速切换。我们展示了它们在各种细胞类型中的适用性以及跟踪抗炎药治疗后激活的鼠巨噬细胞中NO波动的效用。我们期待 1a 和 2a 通过跟踪其在健康和疾病中的动态，将提供有前途的工具来研究NO病理生理学。