Identification and quantitation of B and T cells by cytofluorographic analysis

With cytofluorographic analysis (CFGA) of cells stained with the fluorescent dye acridine orange (AO), the major peripheral white cell populations--lymphocytes, monocytes, and polymorphonuclear cells--display different characteristics and appear as distinct populations which can be quantitated. In this study we present a method for determining percentages of human T and B cells lymphocyte subpopulations by CFGA and display of the data on a computer-generated 3-dimensional grid. Lymphocytes were depleted of either B, T, or both B and T cells by rosetting with erythrocytes and separated by centrifugation. The B cell and T cell depleted and non-rosetting cell subpopulations localized on constant, distinct areas of the display grid. The percentages of T and B cells in peripheral blood samples from 6 normals analyzed by CFGA did not differ from the results obtained by light microscope counting (LMC).