Masson’s Trichrome Stain

Masson’s Trichrome Stain

• Specifications: 5x20mL, 5x100mL (Box)
• Intended use: Intended for the identification of collagen fibers and muscle fibers
• Store temperature: 2℃～8℃

Principle

Masson’s Trichrome Stain method is an adaptation of Mallory’s original trichrome stain, which utilizes a sequence of two or three anion dyes. This method is designed to stain specimens effectively based on differences in molecular weight and tissue permeability. By selecting anion dyes with varying molecular weights and tissue permeabilities, distinct tissue components can be demonstrated.

Main Components

To perform the Mallory’s Trichrome stain, you will need the following reagents:

• Weigert’s iron hematoxylin solution A (containing Hematoxylin)
• Weigert’s iron hematoxylin solution B (containing Ferric chloride)
• Ponceau-acid fuchsin solution (containing Ponceau)
• Phosphomolybdic acid solution (containing Phosphomolybdic acid)
• Aniline blue solution (containing Aniline blue)

Sample Requirement

Ensure that the tissue section is fully fixed before proceeding with the staining procedure.

Test Procedure

Follow these steps to perform the Mallory’s Trichome stain:

• Fix tissues using Bouin’s fixation solution or Zenker’s solution, rinse with running water overnight, and prepare slices through routine dehydration.
• Dewax the slices to water and wash once with distilled water.
• Stain the slices in Weigert’s Iron Hematoxylin Solution (A and B mixed) for 5-10 minutes. Wash briefly with running tap water.
• Briefly place the tissue in 1% hydrochloric acid ethanol, followed by a few minutes of running tap water rinse.
• Stain in Ponceau-Acid Fuchsin Solution for 5-10 minutes, then rinse briefly in deionized water.
• Treat the tissue with Phosphomolybdic Acid Solution for about 5 minutes and pour the solution out without water washing.
• After 3-5 minutes with Aniline Blue Solution, pour out the dye solution without washing.
• Rinse with 1% glacial acetic acid solution and control slides under a microscope to avoid excessive staining.
• Dehydrate with 95% alcohol, proceed with conventional dehydration, transparency, and seal with neutral resin.

Result Interpretation

The Mallory’s Trichrome stain produces distinct color outcomes for various tissue components:

• Collagen fibers, mucus, and cartilage: Blue
• Muscle fibers, cellulose, and erythrocytes: Red
• Nuclei: Purple-black

Precautions:

To ensure successful staining and accurate results, keep these precautions in mind:

• For formaldehyde-fixed tissues, additional fixation in Bouin’s fixative or Zenker’s solution is recommended before staining.
• Mix Weigert’s Iron Hematoxylin Solution A and B in equal parts just before use to prevent oxidation and loss of staining ability.
• Control the treatment time of Phosphomolybdic Acid Solution under a microscope to achieve optimal results.
• Be cautious of excessive decolorization during 95% alcohol dehydration.
• This technique should be performed by professionals who have read the instruction manual, follow hygiene measures, and work within the reagent’s validity period.
• Dispose of waste in accordance with hospital or environmental protection guidelines.
• Check the production date, batch number, and expiration date on the packaging.

References

• Chinese Medical Association. Clinical technical operating specifications-pathology volumes. People’s Military Medical Publishing House.
• Ling Qibo. Practical pathology special staining and histochemistry technology. Guangdong Higher Education Press.

In conclusion, Mallory’s Trichome stain method offers a valuable tool in histological analysis, enabling the visualization of different tissue components through a strategic combination of anion dyes. By following the procedure and precautions outlined above, researchers and pathologists can achieve reliable and informative results in their studies.

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