Based on laser-induced deep etching technology and through-hole filling techniques, TGV interposers were successfully fabricated. The residual stress in the Cu overburden film directly affects the yield improvement after subsequent grinding of the interposer. Therefore, we studied the evolution of residual stress, microstructural changes, and micro-nano mechanical properties within the Cu overburden film after heat treatment at different temperatures. Residual stress analysis using the sin2Ψ method with an X-ray diffractometer (XRD) indicates that during the process of increasing temperature from 100°C to 200°C, the internal residual stress in the Cu overburden film changes from compressive stress to tensile stress. Furthermore, the maximum absolute value of the residual stress, 70.09 MPa, appears at room temperature, and the minimum absolute value of the residual stress, 15.59 MPa, occurs after the heat treatment process at 200°C. Electron backscatter diffraction (EBSD) microstructural analysis shows that the application of heat treatment can effectively accelerate static recrystallization (SRX), promote a higher proportion of high-angle grain boundaries (HAGB), and refine grain size. Additionally, a (001) preferred orientation at high residual stress states and a (111) preferred orientation at low residual stress states are observed. Nanoindentation test results indicate that the Cu overburden film treated at 100°C has the most potential for removal during the grinding process.