Karyotypes of Aedes (Culicidae) mosquitoes (Ae. excrucians, Ae. behningi, and Ae. euedes) have been analyzed using the metaphase chromosomes of imaginal discs. Lacto‐aceto‐orcein, C‐banding, and DAPI staining have detected species‐specific features in the morphology and lengths of these chromosomes in the examined species. Species‐specific features of chromosome 1 in the location of heterochromatin blocks have been shown. Thus, the metaphase chromosomes in the imaginal discs of Ae. excrucians, Ae. behningi, and Ae. euedes are a characteristic for species identification of mosquito species.