Red clover mottle virus (RCMV) was detected by light and electron microscopy in sections of systemically infected pea leaf cells at successive stages of early infection using the combination of a sensitive immunocytochemical method with colloidal gold as a marker of antigen and a low temperature embedding procedure. Gold label was present in the cytoplasm at approximately the same time as the virus-induced membranous inclusions were established in the infected cells as judged by conventional electron microscopy of adjacent tissue. Labelling occurred simultaneously in the cytoplasm and among the membranes of the inclusion but not over the vesicles contained inside the inclusion. Viral antigen was localized by light microscopy with colloidal gold, nominally 8 nm, coupled to protein A and by electron microscopy with colloidal gold, nominally 5 nm, coupled to swine anti-rabbit immunoglobulins. Background staining was low and healthy tissue exposed to specific RCMV antibodies also had a very low level of gold particles.


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