Specimens deposited in Natural History Museum collections are vital for the study of biodiversity, human drivers of biodiversity loss, and other environmental and socioeconomic problems. Despite recent advances in filling our knowledge gaps regarding the myrmecofauna of Cyprus, the island’s biodiversity is considered to be under-sampled. In this article, ant specimens from Cyprus deposited at the Natural History Museums of Crete, the Agricultural University of Athens (Greece), the Muséum d'Histoire Naturelle de Genève (Switzerland), the Royal Belgian Institute of Natural Sciences (Belgium), and the Lund University Biological Museum (Sweden) are examined, presenting a total of 163 specimens belonging to 24 species examined. Stigmatomma denticulatum Roger, 1859 (Amblyoponinae), Myrmecina graminicola (Latreille, 1802), Stenamma debile (Foerster, 1850) (Myrmicinae), and Cryptopone ochracea (Mayr, 1855) (Ponerinae) are reported for the first time for the island of Cyprus, raising its known ant diversity from 77 to 81 species, while additional distributional data are given for species with only a handful of distributional records, i.e. Tapinoma festae Emery, 1925 (Dolichoderinae), Aphaenogaster subterraneoides Emery, 1881 (Myrmicinae), and Hypoponera eduardi (Forel, 1894) (Ponerinae).