A Method for the Quantification of Cellular Composition in Lymphoma Biopsies

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Abstract
A method for the classification and quantification of lymphoma cells in [human] non-Hodgkin lymphomas is described. Lymphoma-cell types were defined and used in differential counts of hematoxylin-and-eosin-stained, paraffin-embedded sections. Cell types were distinguished as follows: small lymphoid cells (small lymphocytes and small plasma cells), medium-sized lymphoid cells (round, cleaved and cerebriform lymphocytes and medium-sized plasma cells) and large lymphoid cells (lymphoblastic, round triple-nucleolated and pleomorphic types). The intra-observer agreement in the classification of cell types was 43-85%. The agreement in distinguishing small, medium-sized and large lymphoid cells was 85-92%. In differential counts, in contrast to conventional histologic classification, all cellular profiles were classified regardless of possible tangential cutting. Of the cellular profiles at different sectional planes of the same cells, 92% were consistently classified, indicating that tangential cutting of cells only influenced the differential counts to a minor degree. By quantification of cellular composition in multiple, randomly selected areas, classified as representative at low power magnification, only small intra-lymphoma variations were observed, in contrast to marked inter-lymphoma variations. The high reproducibility of lymphoma-cell types in combination with the small intra-lymphoma variation makes the quantifiable method suitable in the histological evaluation of non-Hodgkin lymphomas.