Differentiation of human T and B peripheral blood lymphocytes by high-resolution cell image analysis

Automated cell image analysis of light and electron microscopic pictures was used for differentiation of nonlabeled lymphocytes in blood smears and in smears of purified lymphocyte suspensions. The percentages of T and B lymphocytes were determined by a two-step rosette assay with sheep red blood cells (T cells) and an immunofluorescence assay with FITC-labeled antihuman globulin (B cells). Images from 1,400 Feulgen-stained and 12,000 Pappenheim-stained cells were analyzed. Various classification methods allowed two lymphocyte subpopulations to be discriminated at the light and electron microscopic levels on the basis of different visual and subvisual morphologic features. As found by immunologic methods, morphologically determined subpopulations corresponded to T and non-T cells, with no further differentiation of non-T cells into B or null cells possible. The results allow the conclusion that there are morphologic differences between human T and non-T cells, with the differences distinguishable from individual variations as well as from alterations induced by sample preparation.