This study used three staining methods including Hematoxylin-eosin (HE),Wright,and Rapid Gram to compare the staining background,process and effect of plateau zokor (Eospalax baileyi) vaginal smears to screen the best staining method.The results showed that HE staining produced clear smear background and cell types with the nuclear and cytoplasmic staining being significantly different,and high detection rate which was statistically different from the other two methods (P<0.01).Nucleocytoplasmic staining by Wright was blue-purple in varying degrees with messy background,and the difference between nuclear and cytoplasmic staining was not obvious.Rapid Gram staining stained the nucleus and cytoplasm in purple and rose red,respectively,with a relatively dark background,while leukocytes were stained in deep red,with nucleus and cytoplasm not easily distinguished.In conclusion,HE staining could accurately and efficiently identify the estrus cycle of plateau zokor;therefore it is an ideal method for staining vaginal smears.