Identification and isolation of subpopulations of pleural cells by multiparameter flow cytometry

Abstract

Multiparameter flow cytometry was used to identify and sort subpopulations of cells from pleural cell populations harvested from the rat without employing special stains or fluorochrome-labeled monoclonal antibodies. Cell parameters measured included electronic volume, axial light loss, 90 degrees light scatter, and blue autofluorescence. Various bivariate combinations of these parameters were used to distinctly resolve pleural macrophages, eosinophils, mast cells, and lymphocytes. These subpopulations were separately sorted viably according to their unique electrooptical phenotypic characteristics in greater than 90% purity. Our multiparameter flow cytometric approach, accordingly, provides a means by which pleural cell subpopulations may be easily obtained for subsequent in vitro study. Moreover, the general strategy for identifying and isolating these subpopulations may be usefully extended to the identification and isolation of subpopulations of cells occurring in other complex cell mixtures.