Polytene chromosome preparations were obtained from larval, pupal and adult female Malpighian tubules of Aedes aegypti
. The Malpighian tubules of the pupae (0-4 h old) from larvae reared at 20°C provided the best cytogenetic analysis. The interaction of nucleic acids and proteins that influence the spreading of the chromosomes could be reduced with the preparation technique of the sheets submitted to a stronger treatment starting with the hypotony of tissue and successive bathings with acetic acid. A simple technique should facilitate molecular cytogenetics used in the location of resistance and vector competence genes.