PURPOSE: The anterior surface of the cornea of mammals, including humans, has numerous folds in the anterior epithelial cell membranes in the form of microvilli and microplicae. The role of these surface irregularities may be to increase cell-surface area and therefore aid in intra- and extracellular movement of nutritional and waste products across the cell membranes in addition to stabilizing the corneal tear film. The aim of this study was to investigate and compare the nature of these corneal-surface features in various vertebrate classes residing in different environments. METHODS: The anterior corneal surfaces of various vertebrates were investigated by using field emission scanning electron microscopy. Cell areas were analyzed by using image-analysis software. RESULTS: Representative species were examined from all the vertebrate classes, with the exception of the Cephalaspidomorphi. The mean epithelial cell density of aquatic vertebrates (17,602 +/- 9,604 cells/mm2) is greater (p = 0.000018) than that of aerial and terrestrial vertebrate species, including amphibians (3,755 +/- 2,067 cells/ mm2). Similarly, the mean epithelial cell density for the marine vertebrates (22,553 +/- 8,878 cells/mm2) is greater (p = 0.0015) than that of the freshwater and estuarine species (10,529 +/- 5,341 cells/mm2). The anterior corneal surfaces of all species examined were found to show a variety of cell-surface structures. Microvilli are predominant in reptiles, birds, and mammals; microridges appear to be characteristic of the Osteichthyes; and microholes were observed only in the Chondrichthyes. CONCLUSION: The function of these morphologic variations in surface structure appear to be correlated with the range of ecologic environments (marine, aerial, and terrestrial) occupied by each species, corneal phylogeny, and the demands placed on the cornea to ensure clear vision.