The invention discloses a preparation method of a skin microscopic tissue slice of hynobiidae animals and belongs to the technical field of microscopic tissue slices. The preparation method includes following steps: 1) dehydration: successively soaking skin tissue, which has been subjected to fixing treatment, of the hynobiidae animals into ethanol solutions being 50%, 70%, 80%, 90%, 95% and 100% in concentration gradient to dehydrate the skin tissue, wherein dehydration time in each concentration gradient is 45-60 min; (2) transparent treatment: soaking the dehydrated skin tissue into a mixed liquid of 100% ethanol and dimethylbenzene for 2-3 min, and then soaking the dehydrated skin tissue into dimethylbenzene for 10-30 min; (3) wax soaking: soaking the skin tissue being subjected to the transparent treatment into a mixed liquid of dimenthylbenzene and paraffin for 20-30 min, and then soaking the skin tissue into paraffin respectively by two times, each time lasts for 45-60 min; and (4) embedding the wax-soaked skin tissue, and performing slice cutting, slice expansion, slice pasting, slice baking, dyeing and seal-fixation to obtain the skin microscopic tissue slice. The preparation method is simple in process and convenient to operate. The prepared slice is complete and shows a clear image structure.