Silver enhancement of protein A-gold probes on resin-embedded ultrathin sections

Summary A simple method is described allowing the enhancement of the visibility of small gold probes for the electron microscopy.This method, which allows the silver intensification of gold directly on epon-embedded ultrathin sections, was used for the electron microscopic localization of Mouse Mammary Tumor Virus (MMTV) antigens in cultured cells derived from GR and BALB/cfRIII mouse mammary tumors. After the immunostaining with the preembedding protein A-gold technique, the ultrathin sections, placed on 200 mesh copper grids, were rehydrated and exposed to a photographic developer containing silver nitrate. During this physical development gold particles are incapsulated in growing shells of metallic silver, which gradually become more and more visible. We were able to obtain a heavy labelling of the viral particles, well visible even at low magmfication, with a negligeable background staining.The present technique can be useful whenever it is necessary to use the smallest gold probes today available.Supported by contract No. 85.02038.44 from the National Research Council, Rome, Progetto Finalizzato Oncologia