Two-dimensional (2D) materials exhibit exceptional physical and chemical properties owing to their atomically thin structures. However, it remains challenging to produce 2D materials consisting of pure monoelemental metallic atoms. Here free-standing 2D gold (Au) membranes were prepared via in situ transmission electron microscopy straining of Au films. The applied in-plane tensile strain induces an extensive amount of out-of-plane thinning deformation in a local region of an Au thin film, resulting in the nucleation and growth of a free-standing 2D Au membrane surrounded by its film matrix. This 2D membrane is shown to be one atom thick with a simple-hexagonal lattice, which forms an atomically sharp interface with the face-centered cubic lattice of the film matrix. Diffusive transport of surface atoms, in conjunction with the dynamic evolution of interface dislocations, plays important roles in the formation of 2D Au membranes during the mechanical thinning process. These results demonstrate a top-down approach to produce free-standing 2D membranes and provide a general understanding on extreme mechanical thinning of metallic films down to the single-atom-thick limit.