This communication describes a method to obtain enriched populations of T-cells, B-cells and macrophages. Spleen cells were initially fractionated on nylon wool columns. The nylon wool adherent fraction was removed by mechanical agitation and further separated on the basis of adherence to a coated-plastic surface in the presence of autologous serum. The tissue flask adherent population was removed with the aid of a rubber policeman. The nylon wool non-adherent and the tissue flask non-adherent and adherent fractions were characterized for the presence of cell surface markers, size, and functional activity and were identified as T-cells, B-cells and macrophages, respectively. The two-stage adherence procedure is simple to perform and does not require sophisticated equipment or expensive reagents.