Two-dimensional (2D) nanosheets have been assembled into various macroscopic structures for wide engineering applications. To fully explore their exceptional thermal, mechanical, and electrical properties, 2D nanosheets must be aligned into highly ordered structures due to their strong structural anisotropy. Structures stacked layer by layer such as films and fibers have been readily assembled from 2D nanosheets due to their planar geometry. However, scalable manufacturing of macroscopic structures with vertically aligned 2D nanosheets remains challenging, given their large lateral size with a thickness of only a few nanometers. Herein, we report a scalable and efficient microfluidics-enabled sheet-aligning process to assemble 2D nanosheets into a large-area film with a highly ordered vertical alignment. By applying microchannels with a high aspect ratio, 2D nanosheets were well aligned vertically under strong channel size confinement and high flow shear stress. A vertically aligned graphene sheet film was obtained and applied to effectively improve the heat transfer of thermal interfacial materials (TIMs). Superior through-plane thermal conductivity of 82.7 W m-1 K-1 at a low graphene content of 11.8 vol% was measured for vertically aligned TIMs. Thus, they demonstrate exceptional thermal management performance for switching power supplies with high reliability.