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It is generally believed that the introduction of the 3D structure in Cu current collector (CuCC) is beneficial for the uniform deposition of Li due to the reduced local current density and mitigatory reaction rate. However, the influence of the microstructure change in CuCC on the nucleation behaviors of Li through mechanical treatment has been ignored. Hence, microindent is used to prepare 3D structure on the surface of CuCC, the structure evolution during microindentation as well as its impact on Li nucleation behaviors are investigated. It is found that the increased specific surface area results in decreased current density and reduced nucleation driving force. The crystal misorientation beneath the indent leads to the increased surface energy and inhomogeneous nucleation of Li. Thus, the indented CuCC (Ind CuCC) exhibits higher nucleation overpotential and slower electron transfer rate. This manuscript provides a new perspective to the study of the impact of the microstructure of current collector on the nucleation behaviors of Li. [ABSTRACT FROM AUTHOR]