Immunophenotyping with monoclonal antibodies to leucocyte differentiation antigens has an established diagnostic role in the laboratory investigation of acute leukemia. In the vast majority of cases, a hemopoietic lineage can be confidently assigned; namely, acute myeloid leukemia (AML), or the precursor-B and precursor-T variants of acute lymphoblastic leukemia (ALL). The areas of greatest practical importance are in morphologically difficult or undifferentiated cases, and in distinguishing between the major variants of precursor-B and T-ALL. Cases with aberrant patterns of marker expression (acute mixed lineage leukemia, lineage infidelity) are frequently encountered in both ALL and AML, and can lead to diagnostic confusion. However, correlation with morphology and other clinicopathologic features, and careful consideration of the weight of phenotyping evidence almost always allows the correct lineage to be identified. The prognostic value of phenotypic information in acute leukemia is generally limited. Recognition of the major variants of ALL is still of clinical importance, but the significance of myeloid antigen positivity in ALL is controversial, and may not have prognostic value. Patterns of myeloid antigen expression in AML have limited prognostic significance, while the relationship between lymphoid antigen expression and treatment response in AML remains highly controversial. Careful evaluation of the predictive power of immunophenotype in large controlled clinical trials in acute leukemia is still required.