Formalin-fixed and paraffin-embedded patient tissue samples were obtained using the University of Calgary Research Ethics Board protocol REB14-1965. Hematoxylin and eosin staining was carried out as per standard protocol by the Alberta Precisions Laboratories (Calgary, Canada). Immunohistochemistry was performed on the slides as described.10 (link) In brief, endogenous peroxidase activity was inhibited with 3% hydrogen peroxide. The slides were then blocked with 4% goat serum before being incubated with a primary mouse antibody against SARS-CoV-2 (GeneTex GTX632604, Clone1A9) overnight at 4°C. After washing with phosphate buffer solution (5 times, 5 min each), the slides were incubated with biotinylated goat anti-mouse IgG secondary antibody overnight (BA-9200-1.5, Vector Laboratories). Chromogen readout was achieved using the avidin/biotin peroxidase system (PK-6100, Vector Laboratories) and NovaRed substrate (SK-4805, Vector Laboratories).
Protocol detail
Immunohistochemical Detection of SARS-CoV-2 in Formalin-Fixed Tissues
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Anti igg
Antibody
Avidin
Biotin
Buffer
Chromogen
Eosin
Formalin
Goat
Hematoxylin
Hydrogen 3
Immunohistochemistry
Mouse
Paraffin
Patient
Peroxidase
Peroxide
Phosphate
Sars cov 2
Serum
Tissue
Vector
Corresponding Organization : University of Calgary
Other organizations : University of British Columbia, Foothills Medical Centre
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Variable analysis
independent variables
- Incubation of tissue samples with primary mouse antibody against SARS-CoV-2 (GeneTex GTX632604, Clone1A9)
- Chromogen readout using the avidin/biotin peroxidase system and NovaRed substrate
- Formalin-fixed and paraffin-embedded patient tissue samples
- Hematoxylin and eosin staining
- Inhibition of endogenous peroxidase activity with 3% hydrogen peroxide
- Blocking with 4% goat serum
- Incubation with biotinylated goat anti-mouse IgG secondary antibody
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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