The invention relates to a preparation method of a transferable film with a surface micro-structure. The preparation method comprises the steps of carrying out film coating by taking hydroxylated glass as a substrate, using suspension emulsion of polystyrene spheres as film coating liquid and adopting vertical impregnation evaporation self-assembly, so as to form polystyrene sphere colloidal crystal on the glass substrate as a template layer; preparing a corresponding film by taking the template layer as a substrate and adopting a low-temperature magnetron sputtering way, and then carrying out heat treatment on the obtained film at high temperature to remove a template; respectively preparing a polyurethane adhesive layer and a transparent polyester polyethylene terephthalate (PET) layer on the surface of the film, and curing to strip the film from the glass substrate. The transferable film can be prepare into a sticky film by preparing an adhesive layer on the surface of the micro-structure of a film body and sticking a release type protective film . The method can be used for preparing the transferable film with the surface micro-structure; with the transferable property, the film has more flexibility when being applied, so that the application field and application occasion of the film are expanded.