An Immunohistochemistry Staining for the Detection of Rabies Virus Antigens

Begin with a slide carrying a formalin-fixed, paraffin-embedded rabies virus-infected mouse brain tissue section. The fixed section retains viral antigens via protein cross-linking.

Treat the slide with a xylene solution to remove the paraffin.

Immerse the slide in decreasing ethanol concentrations to rehydrate the tissue, making it compatible with aqueous solutions.

Introduce a protease-containing buffer to disrupt protein cross-links, improving antigen accessibility.

Submerge in hydrogen peroxide to block endogenous peroxidases, and apply a blocking solution to prevent non-specific interactions.

Introduce primary antibodies specific to the rabies antigen, forming antigen-antibody complexes.

Add biotinylated secondary antibodies, which bind to the primary antibodies. Incubate with streptavidin-conjugated peroxidase enzymes, which interact with biotin.

Introduce a peroxidase substrate, producing a magenta-red precipitate.

Counterstain the tissue with a nuclear  stain, followed by a bluing agent, staining the nuclei  blue.

The immunohistochemistry image reveals magenta-red spots in the cytoplasm, confirming the presence of rabies antigens in the tissue section.