Abstract The distribution of the threatened ant Dinoponera lucida is limited to the south of the State of Bahia, farthest east of Minas Gerais and north of Espirito Santo, Brazil. Recent cytogenetic studies carried out in 15 sites distributed along the range of the ant indicated high chromosome number variation in the populations of Bahia, 2n=106 to 120 chromosomes, while the populations of Espirito Santo presented a constant number, with 2n=118 chromosomes. This study aimed at describing the banding pattern of D. lucida chromosomes, in some populations of Espirito Santo and Bahia, applying the Hsc-FA technique (secondary constriction heterochromatin-associated bands by fluorescence using acridine) and identifying the nucleolar organizing region (active NORs) impregnated with AgNO 3 which is especially important for the cytogenetic characterization of a species. The utilization of these two techniques showed positive interstitial markings on the region of the long arm of the pair AMt, similar to those obtained with the CMA 3 fluorochrome and with the FISH technique complementing the cytogenetic data of this species.