Masson's Trichrome Stain

Masson Trichrome Stain
Masson Trichrome Stain

Assay-kits

Masson's Trichrome Stain

  • Product code: ACMBS-23
  • Specifications: 5x20mL, 5x100mL (Box)
  • Intended use: Intended for the identification of collagen fibers and muscle fibers
  • Store temperature: 2℃~8℃
  • Expiry date: 18 Months

More Details

PrincipleMain componentsSample requirementTest procedureResult interpretationPrecautions

Principle

This method is modified from Mallory's Trichome stain, which uses the mixture of two or three anion dyes or its function in sequence to stain the specimen. Staining results can be affected by the molecular weight of anion dyes and tissue permeability. Demonstration of different tissue components can be made from selection of anion dyes with different molecular weight and tissue permeability.

Main components

Reagent compositionMain ingredient
1. Weigert's iron hematoxylin solution AHematoxylin
2. Weigert's iron hematoxylin solution BFerric chloride
3. Ponceau-acid fuchsin solutionPonceau
4. Phosphomolybdic acid solutionPhosphomolybdic acid
5. Aniline blue solutionAniline blue

Sample requirement

  • The tissue section must be fully fixed.

Test procedure

  • 1. Tissues are fixed in Bouin's fixation solution (sold separately) or Zenker's solution (sold separately), rinsed with running water overnight, and prepared slices by routine dehydration.
  • 2. The slices are routinely dewaxed to water and washed once with distilled water.
  • 3. Stain in Weigert's Iron Hematoxylin Solution (Weigert's Iron Hematoxylin Solution A and B than a mixed) for 5~10 minutes. Wash briefly in running tap water.
  • 4. Place tissue in 1% hydrochloric acid ethanol for a few seconds. Wash in running tap water for a few minutes.
  • 5. Stain in Ponceau-Acid Fuchsin Solution for 5~10 minutes. Rinse briefly in deionized water.
  • 6. The Phosphomolybdic Acid Solution was treated for about 5 minutes, and the Phosphomolybdic Acid Solution on the slide was poured out (without water washing).
  • 7. After 3~5 minutes of with Aniline Blue Solution, the dye solution on the slide was poured out (without washing).
  • 8. Rinse with 1% glacial acetic acid solution (1% glacial acetic acid solution: 99ml distilled water plus 1ml glacial acetic acid), and slides no blue out (if necessary, under control of the microscope).
  • 9. Followed by dehydration in 95% alcohol for a few seconds, conventional dehydration and transparent, sealed with neutral resin.

Result interpretation

Collagen fibers, mucus and cartilageBlue
Muscle fibers, cellulose and erythrocytesRed
NucleiPurple black

Precautions

  • 1. If the tissue is fixed with formaldehyde solution, the section can be deparaffinized to water and then placed in Bouin' fixative or Zenker's solution for additional fixation for 2~4 hours, rinsed with running water for 10 minutes, be sure to rinse the fixative well before subsequent staining.
  • 2. Weigert's Iron Hematoxylin is divided into solution A and B. Solution A and B should be mixed in equal parts before use. It is not advisable to mix in advance, otherwise it will be easy to oxidize and precipitate and gradually lose staining ability.
  • 3. When Phosphomolybdic Acid Solution is treated, it is best to control it under a microscope. When the muscle fibers are red and the collagen fibers are light red, it is OK.
  • 4. When some tissues are dehydrated with 95% alcohol, it is easy to cause excessive decolorization, which should be controlled by the naked eye.
  • 5. 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.
  • 6. Dispose of waste according to the requirements of the hospital or environmental protection department after use.
  • 7. 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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