The invention discloses a large scale zinc oxide two-dimensional periodic semispherical shell lattice preparation method. The method comprises the following steps: an opal structure template is prepared; an assembly material is grown on a substrate material by adopting a vertical deposition method or a czochralski method to gain a multi-layer or the single-layer colloid crystal and the opal template; the prepared opal template is fixed on a sample holder by a RF high vacuum magnetism control sputtering device and the target material is zinc oxide; the position of the sample holder is regulated in order that the distance between the template and the zinc oxide target conforms to the requirement; a suitable growing condition is selected to get the two-dimensional periodic semispherical shell lattice with different thickness of the large scale zinc oxide. The invention is of simple preparation method, low cost, higher efficiency, high quality of the prepared spherical shell structure and good reproducibility, thus having important application prospect in grating, photo-catalysis and photoluminescence fields.