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In this paper, the effect of annealing condition on the microstructural and mechanical behavior of copper through-silicon via (Cu-TSV) is studied. The hardness of Cu-TSV scaled with the Hall–Petch relation, with the average hardness values of 1.9 GPa, 2.2 GPa and 2.3–2.8 GPa, respectively for the annealed, room temperature (RT) aged and the as-deposited samples. The increase in hardness toward the top of the as-deposited sample is related to the decrease in grain size. The annealed and the as-deposited samples showed a constant elastic modulus (E-modulus) value across the length of Cu-TSV of 140 GPa and 125 GPa respectively, while the RT aged sample showed a degradation in E-modulus from the bottom of the TSV (140 GPa) to the top (110 GPa). These differences in E-modulus values and trends under the different test conditions were found to be unrelated with the crystallographic texture of the samples, but could be related to the presence of residual stresses. No correlation is found between the hardness and E-modulus data. This is attributed to the coupling and competitive effects of grain size and residual stresses, with the grain size effect having a dominant influence on hardness, while the presence of residual stresses dominated the E-modulus result.