Ultrastructural characteristics of peptidergic neurons using pre-embedding immunocytochemical methods

The immunocytochemical staining technique represents a valuable method for electron microscopic analysis with which to elucidate the ultrastructural characteristics, synaptology, and potential interactions of selectively labeled neuronal systems. The unlabeled antibody peroxidase-antiperoxidase (PAP) technique is one of the most widely used immunocytochemical methods and has been applied to both pre-embedding and postembedding EM procedures. This review provides a detailed protocol for the former method and discusses advantages and disadvantages associated with the use of both approaches. A descriptive fine structural analysis of peptidergic neurons localized in the hypothalamus of brain illustrates the applicability and specificity of utilizing this method for the localization of cellular antigens of other systems.