Immunogold labelling of neuroendocrine peptides with special reference to antibody specificity and multiple staining techniques

Abstract

Immunogold methods have been very important for research on the neuroendocrine system. The compatibility of immunogold probes with optimal contrasting for electron microscopy has made localizations of neuroendocrine peptides to different subtypes of secretory organelles possible and, currently, methods using covalent attachment of nanogold particles to antibodies and neuropeptide ligands hold promise for immunocytochemistry, receptor localizations and in situ hybridizations. Multihormonal phenotypes are a hallmark of both the developing and mature neuroendocrine system. The possibility to localize multiple coexisting messengers by multilabelling immunogold methods is emphasized, and different methods for achieving this are discussed. The most difficult part of immunocytochemistry involves definitions and interpretations of specificity, and a number of limitations and control procedures are discussed.