Silver nanoparticle (Ag NP) pastes become a potential die-attachment material with the increased electronic power density. However, the weakness of bonding interface between sintered Ag NPs and bare Cu substrate limits the applications of the Ag NPs paste, thereby reducing the shear strength of the sintered joint. In this work, ultrafast laser processing is utilized to enhance the bonding strength of the sintered Ag joint by fabricating a microstructure interface. The microstructure dimensions are tunable by controlling laser parameters, and then high-strength joints could be obtained. Different substrate microstructures were constructed, and the enhanced bonding mechanism was analyzed by characterizing the cross section and fracture surface morphologies of joints. The ultrafast laser processing could increase the surface energy of Cu substrates to form a more reliable connection with Ag NPs and more energy required for crack extension with the increasing connection area, thereby resulting in a significant improvement in the shear strength of the Ag NP joints. The patterned microstructures on the Cu substrate using this technique showed improved surface energy and increased number of connection areas on the substrate, showing potential for the use in third-generation semiconductors for highly reliable packaging.