Hemipenial morphology has provided useful characters to improve species identification and phylogenetic relationships in squamates. Here we provide hemipenial description and illustration of 13 South American anoles. At generic and specific levels, differences are mainly related to shape and ornamentation; intraspecific variation is low. An asulcate process, present in the hemipenis of most anole species studied, was highly variable among species and may be a useful taxonomic character in hemipenial morphology of this group.