Flow cytometry sorting of unlabelled epidermal Langerhans cells using forward and orthogonal light scatter properties

A variety of techniques based on the presence of specific markers has been proposed to enrich Langerhans cells from epidermal cell suspensions. Computer analysis of multiparameter flow cytometry records involving forward angle and orthogonal scattered light and immunofluorescence of epidermal cells allowed us to determine the scattering properties of Langerhans cells. Unlabelled cells sorted according to these properties were shown to be Langerhans cells by electron microscopy and/or subsequent labelling by anti-HLA-DR monoclonal antibody. The relevance of this method is discussed to sorting viable Langerhans cells which may be used in functional studies and for establishing long-term culture.