Palicourea is a Neotropical genus of predominantly shrubs and subshrubs, represented by more than 700 species, with its greatest diversity in South and Central America. The present study aims to describe and compare the leaf anatomy and micromorphology of six species occurring in the Atlantic Forest to determine which characteristics are most useful for taxonomic study and which are more useful for ecological investigation. Plant material was collected at Atlantic Forest sites in southeast Brazil. Samples preparation followed standard techniques for light and scanning electron microscopy and histochemical tests. The general characteristics observed for genus were absence of domatia, presence of unicellular trichomes on the abaxial surface, hypostomatic leaf blade with paracytic stomata, uniseriate epidermis with papillae, presence of idioblasts, and dorsiventral mesophyll. The results allowed us to segregate species by leaf structure and infer the affinity relationship among the studied species. This study showed the importance of leaf anatomy as a diagnostic tool for the six species of Palicourea; emphasizing characteristics that indicate infrageneric groups.