Virus Stain (Orcein Method)

Virus Stain (Orcein Method)
Virus Stain (Orcein Method)

Assay-kits

Virus Stain (Orcein Method)

  • Product code: ACMBS-34
  • Specifications: 4x20mL (Box)
  • Intended use: Used for staining virus in histological section
  • Store temperature: 2℃~8℃
  • Expiry date: 12 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationPrecautions

Principle

Orcein is weak acid, and alcohol could dissolve it. When dissolving in an acidic solution, it will become dark orange-red. Orcein in acidulated alcoholic solution could stain HBsAg (Hepatitis B surface antigen) is the surface antigen of the hepatitis B virus (HBV) well. When disulfide bond (-SS-) in cystine of HBsAg combining the Orcein solution, it is displayed in colors. Disulfuric bond was oxidized by acidulated potassium permanganate solution into a sulfonic acid. Sulfa group in the sulfonic acid has a strong affinity with orcein and their both combinations were displayed in colors.

Main components

Reagent compositionMain ingredient
1. Potassium permanganate solutionPotassium permanganate
2. Sulfuric acid solutionSulfuric acid
3. Oxalic acid solutionOxalic acid
4. Acidified orcein ethanol solutionOrcein

Sample requirement

  • The tissue section must be fully fixed.

Test procedure

  • 1. The slices are routinely dewaxed to water and washed once with distilled water.
  • 2. Mixed potassium permanganate solution and sulfuric acid solution 1:1 to stain the slice for 5 minutes, then slightly rinse the tissue section with distilled water.
  • 3. Bleach the slice in oxalic acid solution for 2~3 minutes, until it becomes colorless.
  • 4. Rinse with running water 2~3 minutes, and put it into 75% ethanol to wash slightly.
  • 5. Placed it into Acidified orcein ethanol solution for 3~5 hours.
  • 6. Dip in 75% alcohol solution 10~30 seconds twice, until there is no dye precipitate.
  • 7. Conventional dehydration and transparent, sealed with neutral resin.

Result interpretation

  • HBsAg was stained reddish brown to dark brown, with fine granular or homogeneous form. The positive material may form a circular or oval lump, that can also fill the entire cytoplasm, ring-shaped or crescent around the nucleus, Sometimes visible at the edge of the liver cell cytoplasm, showed a lacey. Elastic fibers are also stained dark brown, the remaining background is relatively pale gray-brown. Liver nuclei was not colored.

Precautions

  • 1. After removing the reagent from the refrigerator, please wait until it is fully restored to room temperature before use.
  • 2. Close the lid when staining with acidified orcein ethanol solution, that can be reused several times until the background of the slices too deep to use.
  • 3. This reagent should be used by professionals and interpretation of the results. Read the instruction manual carefully before use, use it within the validity period, and do personal hygiene protection.
  • 4. Dispose of waste according to the requirements of the hospital or environmental protection department after use.
  • 5. The production date, production batch number and expiration date are shown in the outer packaging.

References

1. Chinese Medical Association. Clinical technical operating specifications-pathology volumes [M]. People's Military Medical Publishing House.

2. Ling Qibo. Practical pathology special staining and histochemistry technology [M]. Guangdong Higher Education Press.

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