The invention discloses a preparation method of a brain tissue rapid freezing section. According to the method, acetone is used as a freezing embedding medium and a liquid nitrogen intermediate medium, so that brain tissue can be rapidly frozen , and formation of ice crystals on the brain tissue can be effectively reduced; besides, factors affecting manufacturing quality, such as temperatures during material drawing and sectioning, fixing and soring conditions after sectioning and the like, of the freezing section are optimized, and the quality of the finally prepared section is further guaranteed. According to the brain tissue rapid freezing section obtained with the method, the freezing section which has stability and can well protect antigens and the enzyme activity is provided for further immunofluorescence, immunohistochemistry and in situ hybridization tests on the brain tissue, and a pathological section meeting requirements is provided for future clinical diagnosis related to nervous system diseases and scientific researches. Particularly, molecular and biochemical index detection can be further performed on other tissue of a mouse which is not subjected to perfusion treatment in the scientific research.