In spite of the wide use of electrokaryotypes for Leishmania identification, the number, ploidy and associated functions of the chromosomal bands still remain controversial topics. In the present work, we studied these problems in the pathogenic organism Leishmania mexicana using the clamped homogeneous electric field electrophoresis (CHEF) technique, which allows the separation of uniform chromosomal bands in one run. We arrived at the following general conclusions: (i) a comparative densitometric study using haploid Saccharomyces cerevisiae cells as standard reveals that although L. mexicana is an aneuploid organism, its larger bands are diploid; (ii) a total of 18 chromosomal bands ranging from 3.2 to 0.245 Mbp were resolved. These molecules summed to 1.34 X 10(8) bp, a value within the range of the Leishmania genome; (iii) in hybridisation experiments using different housekeeping gene probes, the majority hybridised with chromosomal band 17 or 18 of L. mexicana, with additional locations for some genes; (iv) the presence of the ubiquitous leishmanial (CA/GT)n sequence in the DNA probes could lead to erroneous gene localisation.