Scanning immunoelectron microscopy for the identification and mapping of two or more antigens on cell surfaces

This study demonstrates the feasibility of using scanning electron microscopy for the identification and mapping of two or more antigens on cell surfaces. Three types of cells: human type B red cells, tumor cells and their associated virus, and guinea pig hepatocarcinoma cells were immunolabeled with ferritin or gold conjugated to IgG fractions of antigen-specific antisera. Gold particles, 300-500 A in diameter, were conjugated to an IgG fraction either directly or indirectly using Staphylococcal protein A. Gold particles of this size can be distinguished from ferritin label using a high resolution scanning microscope. Correlative light, transmission electron microscopy, and scanning electron microscopy studies, as well as x-ray diffraction analysis and the study of stereo micrographs, were performed to visualize the simultaneous immunolabeling by ferritin and gold of two antigens on the guinea pig hepatocarcinoma cell.