Here, we demonstrate for the first time a strategy to self-assemble ZnO nanoparticles (NP) on a large area by a facile one-step process. First, rough and random ZnO nanocrystals (NC), were produced by free-stabilizing aqueous synthesis. Therefore, a post thermal treatment at various temperatures ranging from 80 to 800 °C was necessary to obtain size-tunable and photoluminescent crystalline NP. The fabricated NP had both efficient UV photoluminescence and photocatalytic activity by photo-degradation of Methylene Blue (MB) dye. The annealed NP showed an absorption blue shift in the UV region with decreasing size. This shift was attributed to high quantum confinement effect since ZnO NP diameter reached values lower than the Bohr radius of ZnO (~2.7 nm). The photocatalytic activity displayed dependency on the particle’s size, number, and crystallinity. Subsequently, the NP were self-assembled inside poly(methyl methacrylate) (PMMA) nanoholes. Subsequently, large area substrate of homogenous properties ZnO NP was obtained. Moreover, the synthesis facility, photoemission and photocatalytic properties of ZnO NP could be a new insight into the realization of high performance and low cost UV laser devices.