Immunophenotyping by Multiparameter Flow Cytometry

Multiparameter flow cytometry has become an indispensable tool for the diagnosis and classification of acute myeloid leukemia (AML). The basic method relies on the unique ability to detect immunophenotypic abnormalities on discrete subpopulations. The primary roles in the initial assessment of AML are to determine the immaturity of the leukemic population, define the lineage and the immunophenotypic aberrancies in blasts, and identify characteristic immunophenotypic features to predict important recurrent cytogenetic and genetic abnormalities and prognosis. The established immunophenotypic profile, a baseline “fingerprint,” is used for follow-up assessment of residual disease. This chapter provides an overview of procedures for specimen processing, staining, and immunophenotyping of AML and describes our strategy for data analysis supplemented with illustrative case examples.