Light microscope identification of murine B and T cells by means of functional polymeric microspheres

Abstract

Covalent bonding of purified antibodies to polymeric microspheres of 0.4 to 0.8 μm diameter yields conjugates which can be used to label lymphocytes in the light microscope. Nonadherent microspheres can be separated by means of a discontinuous density gradient and quantitative measurements of adherent microsphere distributions can be made through examination of Wright's stained dry mounts or through fluorescent microscopic examination of cells in suspension. In general the distributions of adherent microspheres on mouse splenic and thymic lymphocytes in direct or indirect labelling assays show good agreement with results obtained from fluorescent antibody techniques. In comparison to fluorescent antibody the use of these antibody-microsphere conjugates has the advantage of allowing direct correlations between the surface antigens of cells and their histologie morphology.