beta-N-Acetylhexosaminidase isoenzymes, and the distribution of the alpha- and beta-subunits forming the enzyme in a representative series of fresh leukaemic cells and in established leukaemic cell lines, were obtained by using a combination of DEAE-cellulose chromatography and assay with the fluorogenic substrates 4-methylumbelliferyl-beta-N-acetylglucosaminide hydrolyzed by both alpha and beta subunits, and 4-methylumbelliferyl-beta-N-acetylglucosaminide-6-SO4 hydrolyzed only by hexosaminidase isoenzymes containing alpha-subunits. The presence of hexosaminidase S (the alpha alpha dimer), was found in all the leukaemic cell populations we surveyed, but not in normal human cells. The presence of this isoenzyme can therefore be considered as an additional marker of leukaemic cells. A prevalence of hexosaminidase A and A-like intermediate forms (alpha beta structure), characterize leukaemic cells of myeloid origin, whereas greater amounts of hexosaminidase B and B-like intermediate forms (beta beta structure), were consistent attributes of leukaemic cells of lymphoid origin. An over-expression of beta-subunits in blasts might be related to their undifferentiated status. These changes in the isoenzymes of hexosaminidase may prove informative about a variety of changes in the biology of leukaemic cells that could range from chromosomal alterations to changes in the proteolytic processing and glycosylation.