A fabricating method of a structure having nano-hole is provided. The fabricating method includes: providing a substrate, forming a photoresist layer on the substrate, forming an opening, and performing a heat treatment process on the photoresist layer to shrink the opening to form a nano-hole. The structure having nano-hole fabricated by the method of the present invention can be used to fabricate a nano-tip having a diameter of tip-body of no more than 10 nm, high aspect ratio, and a uniform diameter of tip-body.