The invention relates to a nanoparticle self-assembled transparent circuit board, a preparation method thereof, and an application thereof, and belongs to the technical field of microelectronics. Thenanoparticle self-assembled transparent circuit board comprises a nanoparticle conductive structure and a hydrophilic transparent substrate, wherein the nanoparticle conductive structure is a hollow structure; the nanoparticle conductive structure is located on the surface of the hydrophilic transparent substrate; and the nanoparticle conductive structures are distributed in an array. The preparation method comprises the following steps: firstly, preparing the hydrophobic seal, paving one side, with the concave part, of the seal on the paraffin film, and heating the paraffin film, so that theparaffin film is converted into a molten state; and setting a seal on a hydrophilic transparent substrate, adhering molten paraffin on the seal to the surface of the hydrophilic transparent substrate,then tearing off the hydrophilic transparent substrate, adding dropwise a nano-particle aqueous solution to the hydrophilic transparent substrate, and forming the transparent circuit board based on nano-particle self-assembly based on the coffee ring effect. The nanoparticle self-assembled transparent circuit board has the advantages of simple and rapid manufacturing, low cost, diversified manufacturing substrates, diversified patterns, high throughput, transparency and the like.