Clinical flow cytometry in molecular genetic pathology

Abstract

Flow cytometry is a powerful diagnostic tool most commonly used for immunophenotyping of hematopoietic neoplasms and primary/secondary immunodeficiencies. Other applications include cell sorting, detection of chromosomal abnormalities based on in situ hybridization or polymerase chain reaction, and evaluation of various cellular functions such as proliferation, apoptosis, calcium efflux, and phosphorylation. Flow cytometry measures physical and antigenic properties of particles including cells, chromosomes, and individual molecules. Owing to the availability of numerous antibodies and ever expanding array of fluorochromes, flow cytometry is capable of simultaneous evaluation of multiple parameters. In hematolymphoid malignancies, flow cytometry can be used at the time of the initial diagnosis and for follow-up and prognostication. The diagnosis is based on aberrant immunophenotypes and maturation patterns, different from those typically seen in normal hematopoietic populations. This chapter focuses on the diagnostic applications of flow cytometry and presents a current approach to sample preparation, selection of antibody panels, and essential aspects of analysis and interpretation. In addition, a complete review of immunophenotypic features of major entities of 2008 World Health Organization classification of hematolymphoid neoplasms is discussed.