This invention is within the field of eye medicine, involving the preparation method of one type of cornea intravital staining. The above mentioned staining includes the mixed solution of fluorescein sodium aqueous solution and lissamine green aqueous solution. In the above fluorescein sodium aqueous solution, the concentration of fluorescein sodium is 0.5%-4.0% w/v; while in the above lissamine green aqueous solution, the concentration of lissamine green is 0.5%-4.0% w/v. And the volume ratio of the above mentioned fluorescein sodium aqueous solution and lissamine green aqueous solution is 1:0.25-1:3. The cornea intravital staining in this invention possesses some advantages like making the staining of cornea and bulbar conjunctiva proceed at the same time, completed by one time, little irritation to eye tissue and being susceptive for patients. And it is mainly used in diagnosing and evaluating xerophthalmia, keratohelcosis, keratitis (KCS), arborized corneal epithelium herpes, and early diagnosis of Sjogren syndrome etc.