Improved Detection of Diffuse Large B-Cell Lymphoma by Flow Cytometric Immunophenotyping-Effect of Tissue Disaggregation Method

Background: Flow cytometric immunophenotyping (FCI) is recognized as a rapid, sensitive, and accurate method for diagnosis of B-cell lymphomas. We observed that FCI failed to identify the clonal B-cell population in several cases of large B-cell lymphoma (DLBCL) when tissue samples were prepared by a commercially available mechanical tissue disaggregation method. We tested a manual tissue disaggregation method and compared it with the mechanical method. Methods: FCI findings from 51 cases of DLBCL processed with the mechanical tissue disaggregation method, 27 cases processed using the manual method, and 15 cases processed using a combination of both methods were compared. The histological and immunohistochemical findings in each case were reviewed. Results: FCI detected a clonal B-cell population in 88.9% of cases processed by the manual tissue disaggregation method, 66.7% of cases processed by a combination of the manual and mechanical disaggregation methods, and in 62.7% of cases processed solely by the mechanical tissue disaggregation method (P<0.01 Fisher exact). Manual processing yielded positive FCI results in 81.8% of the nodal tissue samples and 93.8% of the extra-nodal tissue samples, whereas mechanical disaggregation was particularly inefficient in preserving large lymphoma cells from extra-nodal tissue: 71.4% of the nodal and 56.8% of the extra-nodal tissue samples processed by the mechanical method showed clonal B-cells by flow cytometry (P<0.006, Fisher exact). Conclusions: The diagnostic yield of FCI in DLBCL can be significantly improved by utilizing a manual disaggregation method, particularly in extra-nodal tissue samples. (C) 2015 International Clinical Cytometry Society