Light microscopical detection of single 5 and 20 nm gold particles used for immunolabelling of plasma membrane antigens with silver enhancement and reflection contrast.
To detect plasma membrane antigens, cytocentrifuge preparations of the macrophage-like cell line P388D1 were incubated with monoclonal antibodies and labelled with 5 and 20 nm gold particles conjugated to immunoglobulins or protein A. The 20 nm, but not the 5 nm, particles could be observed by reflection-contrast light microscopy. Single 5 nm particles, however, were clearly demonstrable with silver contrast enhancement, both in absorption contrast and in reflection contrast. The method proved to be compatible with cytochemical methods such as acid phosphatase.
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis*; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology* / Q000648:ultrastructure; D006046:Gold; D007202:Indicators and Reagents; D051379:Mice; D008853:Microscopy / Q000295:instrumentation / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D012834:Silver
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D007202:Indicators and Reagents; D012834:Silver; D006046:Gold