The use of ultrathin cryosections for localisation of influenza virus antigens in infected vero cell cultures

Abstract

The localisation of influenza virus antigens in infected Vero cell monolayer cultures by post embedding immunoelectron microscopy requires both good resolution and the retention of antigenicity in the tissue sections. Ultrathin cryosections are superior to ultrathin resin sections for this purpose. The colloidal gold probe was used in conjunction with specific antibody preparations to localise three viral proteins. Antibody raised against haemagglutinin glycoproteins labelled the host cell membrane and the virus fringe without contamination of the host cell nucleus, whereas antibody raised against viral nuclear protein labelled throughout the host cell cytoplasm and nucleus. Matrix protein was localised within the nucleus and was associated with the host cell membrane of the infected cell. The appearance of all these proteins was maximal 24 h post infection.