The use of immuno-gold silver staining in bluetongue virus adsorption and neutralisation studies

The immuno-gold-silver staining (IGSS) technique was used in scanning electron microscopy for the detection and semi-quantitation of low copy antigens on the surface of cells. The methodology was exploited in experiments designed to examine the interaction of small numbers of virus particles with the surface of susceptible host cells. Using bluetongue virus (BTV) as an example, IGSS procedures confirmed that maximum adsorption occurred within 60 min and that adsorbed virus particles were distributed randomly on the surface of the cell. Neutralising antibody did not prevent binding of BTV to the plasma membrane, but abrogated virus uptake. The use of IGSS in the study of virus-cell interactions was validated by transmission electron microscopy and classical biochemical experiments utilising radioactively-labelled virus.