Nano-structured Ni/Ni3Al multilayer was prepared by magnetron sputtering, with individual layer thicknesses h varying from 10 to 160 nm. The microstructure and hardness of Ni/Ni3Al multilayer were investigated by X-ray diffraction, transmission electron microscopy and nanoindentation. The results show that the hardness increases with decreasing h for as-deposited and 500 degrees C annealed multilayers. When annealed at 700 degrees C, the hardness approach a peak value at h =40 nm with followed by softening at smaller h. The influence of individual layer thickness, grain size as well as formation of ordered Ni3Al on strengthening mechanisms of Ni/Ni3Al multilayers at elevated temperature are discussed. Crown Copyright (C) 2016 Published by Elsevier B.V. All rights reserved.