Applications of immunogold‐silver enhancement: Testing of monoclonal antibodies and detection of human cytomegalovirus in histologic specimens

Abstract

Human cytomegalovirus (HCMV) is an important pathogen in neonates, transplant patients, and individuals with acquired immunodeficiency syndrome (AIDS). Reliable techniques for the detection of this virus in clinical specimens would aid in improving methods for diagnosis and increasing our understanding of viral pathogenesis. We evaluated the utility of immunogold-silver enhancement to determine the specificity of murine monoclonal antibodies directed against HCMV proteins and applied these antibodies to the detection of HCMV in histologic material. Nine antibodies were tested in a tissue-culture system to determine the location of staining. All were found to be active in frozen tissue, but only two of these antibodies were reactive in formalin-fixed tissue. We also evaluated a novel tissue fixative technique (AMeX; Fixation and dehydration with acetone) that has been used to maintain antigenicity of T-lymphocyte cell markers. All antibodies remained reactive against their respective HCMV proteins in tissue fixed by this technique. However, dehydration of tissue may limit the usefulness of AMeX fixation. Immunogold-silver enhancement is a useful and reliable histochemical technique for detection of HCMV antigens in pathologic specimens.

Optimization of differential immunogold-silver and peroxidase labeling with maintenance of ultrastructure in brain sections before plastic embedding

“…Moreover, the reaction was difficult to control based on visual cues due to its light sensitivity (Massari et al, 1988). The availability of smaller 1 nm colloidal gold probes and light insensitive silver intensifiers (Janssen) now suggest that this labeling procedure may be a highly reproducible and sensitive immunocytochemical marker (In t Veld, 1989;Dankner and Spector, 1989). In perfusion-fixed brain tissue, we sought to determine the conditions under which the 1 nm colloidal gold probes and light insensitive silver intensifier might be used to achieve: (1) cellularly and subcellularly selective localization of antisera; (2) optimal preservation of ultrastructure; and (3) differentiation from immunoperoxidase labeling of a second antiserum within the same section.…”

The application of a biotin-anti-biotin gold technique providing a significant signal intensification in electron microscopic immunocytochemistry: a comparison with the ultrasmall immunogold silver staining procedure

“…Previous modifications have included the application of streptavidin gold (Dankner and Spector 1989;Brändli et al 1990) or an anti-horseradish peroxidase antibody-gold complex (Gee et al 1991;Roth et al 1992).…”

Immunohistochemical Detection of Porcine Rotavirus Using Immunogold Silver Staining (IGSS)

“…Streptavidin-gold and silver enhancement 3,4 were used for the detection of cytomegalovirus in both cell culture and histologic specimens. A secondary antibody-gold conjugate was utilized followed by silver enhancement, in the diagnosis of herpes encephalitis, 2 and the results correlated well with those obtained by peroxidase-anti-peroxidase or immunofluorescence methods.…”