This study is based on sampled data obtained in 60 equally distant municipalities of São Paulo State. In this study, we proposed a tool to identify Acromyrmex species in a practical and agile manner through the use of an illustrated key that demonstrates the morphological attributes of workers, associated with the type of cut plant material, external and internal appearance of nests and nidification sites. Through sampling, eleven species and six subspecies of Acromyrmex occurring in the State were identified, including the first registers of A. diasi and A. subterraneus molestans.