An improved method for selecting T-cell hybridomas by the fluorescence-activated cell sorter.

A two-step sorting procedure with the fluorescence-activated cell sorter (FACS) is described to select T cell hybridomas from cell populations obtained from the fusion of activated T cells and T- lymphoma cells. Cells are first sorted on the basis of differences in cell-surface antigens. After a period of growth in culture, the cells are then stained with the DNA-labeling dye dis-benzimidazole 33258 Hoechst and sorted on the basis of their DNA content. From 15 to 25% of the resulting cell clones, growing out of limiting dilutions, were hybrid lines as judged by chromosome analysis.