Many orb-weaving spiders exhibit remarkable sexual dimorphism, hampering the matching of males and females in taxonomic studies. This is the case for the spiny Micrathena spiders, a species-rich Neotropical genus with 27% of its species known from a single sex. In this paper we document several undescribed Micrathena specimens, and test whether they belong to some of those incompletely known species. In order to do so, we: (1) tested the phylogenetic position of males and their putative females using a previous morphological dataset; (2) calculated genetic distances among individuals based on a fragment of the mitochondrial gene cytochrome c oxidase subunit I; and (3) examined their geographical distributions. These approaches, isolated or in combination, allowed us to identify and describe the previously unknown males of M. embira Levi, M. reimoseri Mello-Leitão, M. exlinae Levi, M. miles Simon, M. spinulata F.O. Pickard-Cambridge, M. yanomami Magalhães & Santos and M. cornuta (Taczanowski), and the female of M. beta di Caporiacco. We found that the males previously associated with M. bicolor (Keyserling), M. cornuta and M. lata Chickering had been incorrectly matched with females. The latter actually belongs to a hitherto unnamed species, herein described as Micrathena perfida, sp. nov. New geographical data are given for these and other Micrathena species. Our study highlights the importance of using different sources of data for matching the sexes in diverse groups with strong sexual dimorphism.