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Our wafer-scale atomic layer lithography method can benefit a diverse range of 2D optical and electronic applications.Glutathione (GSH) level in cells maintains redox homeostasis, so the detection of intracellular GSH is crucial for many pathological processes. In the present study, we developed a cystamine-functionalized glass nanopore (cG-nanopore) for the specific detection of GSH in single cells through the Michael addition reaction between the amino group of cystamine and polydopamine adhered to the inner wall of the glass nanopore.