The invention discloses a medical vivisection testing method for a Kunming (KM) mouse, and relates to the technical field of life science. S3, dissecting: a scalpel is utilized to remove calvaria skinbetween both eyes in a direction perpendicular to the nasal bone of the KM mouse, and meanwhile, skin from the parietal region to the occipital bone between both ears is removed to expose skulls, bone forceps are utilized to enter the skulls for certain depth and cut into the head, then, scissors are utilized to cut open the parietal bone, and the skulls are opened; and S4, sampling: the skulls at two sides outside the cerebrum are separated and taken away to expose complete brain tissue, and surgical scissors and ophthalmic forceps are utilized to peel off meninges. According to the medicalvivisection testing method for the KM mouse, the skulls of the KM mouse can be rapidly separated to obtain complete and pollution-free brain tissue samples including the cerebrum, epencephalon, hypophysis, hypothalamus and the like; compared with a traditional method, the medical vivisection testing method improves the collection integrity of all the tissue samples of the brain, and lowers the risk of operation pollution and sample damage; and the medical vivisection testing method is simple and has a good using effect.