The invention discloses a preparation method of a nanopore chip with a plasmon resonance scattering response function. The preparation method comprises the following steps: (1) selecting a chip substrate: selecting a silicon chip as a substrate, and depositing an insulating material on the front surface of the silicon chip, without the insulating material on the back surface; (2) etching a square area on the back surface of the silicon chip by a wet etching method until a nanopore chip film is exposed on the back surface; (3) etching nanopores: forming the nanopores on the nanopore chip film by using a focused ion beam or a focused electron beam to bomb the nanopore chip film; (4) modifying noble metal: coating a noble metal layer on the surface of the nanopore chip film by using a magnetron sputtering instrument. The preparation method is simple in steps; the prepared nanopore chip is controllable in size and relatively low in cost; through noble metal modification on the nanopores, the nanopore chip with a photoelectric response function is prepared; the nanopore chip can cause a plasmon resonance coupling phenomenon so as to amplify a spectral signal, and can be applied to many fields such as electrochemical detection of the nanopores.