| Attribute | Details |
|---|---|
| Cat | G1921 |
| Size | 3 x 50mL |
| Storage | 2-8°C, avoid light, valid for 6 months |
Kit Components
| Reagent | Volume | Storage |
|---|---|---|
| Reagent(A): | ||
| Shikata Acidizing Solution | Al: Shikata Oxidant | 25mL |
| A2: Shikata Acid Solution | 25mL | |
| Reagent(B): | Shikata Bleaching Solution | 50mL |
| Reagent(C): | Acidic Orcein Stain Solution | 50mL |
Before use, mix Al with A2 in equal amounts to form Shikata Acidizing Solution. It is ready to use.
Introduction
- Viral Hepatitis Description:
- Viral hepatitis is an infectious disease characterized by the degeneration and necrosis of liver parenchymal cells induced by hepatitis viruses.
- Prevalence of Hepatitis B Virus (HBV):
- Hepatitis B virus (HBV) is globally prevalent.
- HBsAg (Hepatitis B surface antigen) is typically present in the liver cytoplasm.
- Immunohistochemistry for HBsAg and HBcAg:
- Currently, HBsAg and HBcAg can be displayed via immunohistochemistry techniques.
- Staining Methods for HBsAg:
- Various methods exist for displaying HBsAg, including the Shikata Orcein Stain Method, Aldehyde Fuchsin Method, Victoria Blue Method, etc.
- Kit Description:
- This kit offers simplicity and specificity in staining HBsAg.
- The staining effect may vary with different batches of raw materials.
- Orcein likely combines with the disulfide bond of cystine in HBsAg to produce color.
Protocol (Our reference only)
- Fix the tissue in 10% formalin. Conventionally dehydrate and embed.
- Cut the section into 4μm. Conventionally dewax and rehydrate.
- Oxidize with Shikata Oxidant for 5 minutes. Rinse with water slightly.
- Bleach with Shikata Bleaching Solution for 2 minutes to remove the color of Shikata Acidizing Solution.
- Rinse with distilled water slightly. Immerse with 70% ethanol slightly.
- Stain with Acidic Orcein Stain Solution under cover for 5 hours or more.
- Differentiate with 95% ethanol until a clear light brown base is achieved.
- Dehydrate rapidly with absolute ethanol, transparentize with xylene, and seal with resinene.
Result
- HBsAg Positive Substance: Brown-red to deep brown
- Background: Light red on different levels
Note
- Shikata Acidizing Solution must be used immediately; otherwise, its oxidation power will be reduced.
- Acidic Orcein Stain Solution should be restored to RT before use, and it can be reused several times.
- After staining for 5 hours, remove the section, wash it in 70% ethanol, then wash it in distilled water, and view under the microscope. If the staining is light, soak it in 70% ethanol and then stain with Acidic Orcein Stain Solution. If necessary, it can be stained overnight.
- When staining with orcein, lipofuscin has a positive reaction. The difference is that lipofuscin has a deeper color, presenting small particles of uniform size in the cytoplasm; HBsAg has a shallower color, presenting fine particles.
- For safety and health, please wear laboratory attire and disposable gloves.
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