Considerable structural, metabolic, and proliferative heterogeneity develops in populations of cultured diploid cells which have reached advanced levels of population doubling. Isolation of noncycling cells from late-passage cultures would permit more definitive investigation of the structure and behavior of individual senescent cells. In this paper, we report the viable sorting of late-passage cultures of human diploid fibroblasts (IMR-90) into two subpopulations of cells with different proliferative potentials. Sorting is based on cellular light-scattering properties and autofluorescence. Structural and behavioral features of the subpopulation exhibiting increased forward-angle light scatter are more characteristic of senescent cells than the subpopulation sorted by decreased forward-angle light scatter.