The invention discloses a preparation method of a fungus specimen. The preparation method comprises the following steps of (1) collecting and cleaning up the sporocarp of the fungus which serves as the specimen, wherein the sporocarp is taken as a substrate; (2) immersing the substrate in liquid nitrogen; (3) volatilizing the liquid nitrogen, and then carrying out microwave heating; and (4) preparing transparent epoxy resin AB glue, coating the surface of the substrate obtained after the step (3) with the transparent epoxy resin AB glue, carrying out curing, and obtaining the fungus specimen.According to the method, during the preparation process, the microstructure of the fungus can be completely reserved, and the water content is rapidly reduced while the structure is not damaged, so that the specimen has the relatively long storage life.