It had been shown that nanotwinned Cu (nt-Cu) possess a lot of advantages, including high strength and ductility with a little bit enhancement of electrical conductivity. In addition, it has a great potential to apply on 3-dimensional integrated circuits (3D IC), which is believed to be an advanced method to increase the density of joints in the field of packaging technology with respect to scaling. Thus, many studies have been dedicated to seeking the best conditions to produce nt-Cu with distinguished properties by changing a lot of parameters including current density, films thickness, electrodeposition mode, etc. It had been reported that nt-Cu could be fabricated successfully by pulse-current mode. Besides, it could also be fabricated by direct-current (DC) mode from the previous research . While undergoing the process of electrodeposition, Cu films can form nanotwins because of stress relaxation . Moreover, if we want to judge on the preferred orientation after thermal treatments, film thickness needs to be considered . With different current density, the microstructure of Cu films changes as well . Temperature always play a vital role speaking of materials science, however, there is just little understanding concerning the effect of temperature of electrodeposition. In the present study, we examine the changes of the microstructure with plating temperatures and investigate the thermal properties after annealing processes under different conditions.