Objective To observe the morphological features of mouse cornea after fixation with 6 different types of fixatives, and choose an optimal fixative suitable for fixation of mouse cornea. Methods 60 eyes of NIH mice were randomly divided into 6 groups, 10 eyes in each group. The eyes were respectively fixed in the 6 types of fixatives, namely, 95% ethanol, 10% neutral buffered formalin, 10% formalin, Bouin's solution, Formaldehyde-acetic acid fixative solution, and Davidson's solution. After fixation, the specimens were dehydrated with 80%, 95%, 100% gradient alcohol, and clearing was performed using TO-type histological clearing agent, then soaked in melted wax, and finally embedded in paraffin. The paraffin sections were 4 μm thick, and stained with hematoxyline and eosin. The histological features of the cornea were observed and various regions of the cornea were measured. Results Different fixatives showed different histological characteristics and thickness of the mouse cornea differently. The acetic acid-formaldehyde fixative caused least changes of the morphology and thickness of the mouse cornea.Conclusions It is important to consider the impact of different fixatives on the corneal tissue morphology. In order to avoid histological artifacts, formaldehyde-acetic acid fixative among the 6 examined fixatives is the most suitable for histological examination of mouse cornea.