Flow Cytometry Immunophenotyping of Hematolymphoid Neoplasia

Abstract

Flow cytometry immunophenotyping (FCI) of hematolymphoid specimens is a powerful tool in clinical medicine aiding in the accurate diagnosis and subclassification of acute and chronic leukemias, non-Hodgkin lymphomas, myelodysplasia, and other hematolymphoid neoplastic processes. Multiple flow cytometric strategies are used to evaluate hematolymphoid populations, including identification of neoplastic populations with aberrant immunophenotypes, abnormal maturation patterns, monotypic kappa/lambda light chain expression, restricted V-beta expression, and abnormal light scatter properties. In this chapter, we present a general approach to analyze hematolymphoid populations. Specimen handling, gating strategies, and appropriate antibody panels for flow cytometric detection of abnormal hematologic populations are presented.

Tissue flow cytometry immunophenotyping in the diagnosis and classification of non‐Hodgkin's lymphomas: A retrospective evaluation of 1,792 cases

“…Data acquisition was performed on a FACSCalibur flow cytometer (BD Biosciences) and analyzed by CellQuest Pro software until 2008, or afterward on FACSCanto II (BD Biosciences) using FACSDiva software. Sequential panels were used (Table 1): first, we performed a screening panel in order to detect the presence of lymphoma; if the lymphoma was detected, an additional panel followed to narrow the differential diagnosis according to B/T-cell lineage (8). If epithelial tumors were of concern, a tube containing monoclonal antibody anti-cytokeratin was added.…”

“…It must be considered that among these cases, the majority (20/47) belonged to the first three years of our study, when the multiparameter analysis was still limited as we used a three-color immunophenotyping protocol on a Calibur instrument, so antibody panel selection could have been misdirected and gating strategies would be too restrictive or too inclusive to adequately detect tumor population. Adverse storage conditions, inadequate sampling, or manipulation due to the difficulty encountered in obtaining sufficient cells from certain tissue types could also be invoked to explain false-negative results (8). Moreover, many neoplastic nodes or masses can show dishomogeneous involvement and inaccurate sampling can lead to a biopsy which is devoid of malignant cells and not representative of total mass (59).…”

Flow cytometry identification of nonhemopoietic neoplasms during routine immunophenotyping

“…CD45 (leucocyte common antigen) is a sort of lighthouse for cytometrists involved in leukaemia/lymphoma diagnosis, as its expression allows the recognition of the leucocyte populations and deviation from normal pattern is highly indicative of a pathological condition . Among haematological diseases, CD45 may result negative in some acute leukaemias (B‐lymphoblastic and megakaryoblastic) and in multiple myeloma . Apart from these entities, detection of CD45− cells is a strong clue for the presence of a NHNs, mostly if associated to positivity of other selected antigens, including CD56, expressed by neuroendocrine tumours .…”

Visualization of Cell Composition and Maturation in the Bone Marrow Using 10‐Color Flow Cytometry and Radar Plots

“…The first papers on normal patterns of differentiation in hematopoietic cells were published by Terstappen et al in 1990s using five parameters (22)(23)(24) and were followed by other publications (25)(26)(27)(28)(29)(30)(31)(32)(33)(34). The first papers on normal patterns of differentiation in hematopoietic cells were published by Terstappen et al in 1990s using five parameters (22)(23)(24) and were followed by other publications (25)(26)(27)(28)(29)(30)(31)(32)(33)(34).…”