Immunoelectron microscopy and immunocytochemistry in pathology, with special reference to immunoglobulin-producing...

The advantage of immunoelectron microscopy (immuno-EM) is that it allows the simultaneous detection of surface and internal cell components. The immunoperoxidase method is often more suitable than immunoferritin. There are no major difficulties in staining surface antigens by immuno-EM, provided sufficient amounts of pure antibodies are available for coupling to peroxidase. Prior fixation of cells, with faxatives that preserve the antigenicity of surface components, avoids ligand-induced alterations of the surface components. It is believed that, unlike the surface, intracellular antigens are difficult to stain by immuno-EM because of the poor penetration of conjugates into fixed cells; thus, various technical approaches have been proposed by workers involved in tissue immuno-EM. In fact, the method that we initially devised for the surface staining of fixed cell suspensions has proved to detect specifically intracellular immunoglobulins in B cells obtained from patients with proliferative diseases. Thus, conjugates do penetrate into fixed cells, although by an unknown mechanism. On this basis, it is possible to study both surface and intracellular immunoglobulins at the EM level and to determine the precise localization synthesis.