The　thermomechanical　reliabilities　of　TSV　based　structures　are　closely　affected　by　the　protrusion　deformation,　stress-strain　constitutive　behavior　and　microstructure　evolution　of　TSV-Cu　under　various　thermal　loadings.　In　this　study,　the　stress-strain　constitutive　curves　of　TSV-Cu　under　various　annealing　conditions　considering　holding　time　effect　are　acquired　by　the　nanoindentation　test　and　finite　element　(FE)　reverse　analysis.　The　yielding　softening　in　stress-strain　curves　is　found　if　the　holding　time　is　less　than　90　min.　However,　the　stable　stress-strain　curves　are　obtained　as　holding　time　exceeds　90　min.　Furthermore,　the　mechanisms　of　softening　and　stability　in　stress-strain　curves　are　revealed　by　observing　the　microstructure　evolution　of　TSV-Cu　with　a　wide　range　of　holding　time.　In　addition,　the　relationships　among　protrusion　height,　constitutive　curves　as　well　as　micro-structure　parameter　of　TSV-Cu　with　different　annealing　holding　time　are　built.　This　study　provides　an　intrinsic　insight　on　the　correlations　of　deformation,　constitutive　curves,　and　microstructure　for　TSV-Cu　annealing　pro-cesses　considering　holding　time　effect.