Masson's Trichrome for Connective Tissue
EMS Catalog #: 26367-Series
Fixation:
Bouin’s (#15990-01) or 10% Buffered Formalin (#15741-01)
Sections:
Paraffin at 6µm
Staining Procedures:
- Deparaffinize and hydrate to distilled water.
- Mordant in Bouin’s Fixative (#26367-01) for 1 hour at 56°C or overnight at room temperature.
- Cool and wash in running water until yellow color disappears.
- Rinse in distilled water.
- Stain in Weigert’s Iron Hematoxylin Working* solution for 5 minutes. Wash in running water for 10 minutes.
- To prepare working solution, mix equal parts of Weigert’s Iron Hematoxylin A (#26367-02) and Weigert’s Iron Hematoxylin B (#26367-03).
- Rinse in distilled water.
- Place in Biebrich Scarlet-Acid Fuchsin (#26033-25) for 15 minutes.
- Rinse in distilled water.
- Place in Phosphomolybdic Acid-Phosphotungstic Acid (#26367-05) for 15 minutes.
- Stain in Aniline Blue Solution (#26367-06) for 10 to 20 minutes.
- Rinse in distilled water.
- Differentiate in Acetic Acid, 1% (#26367-07) for 3 to 5 minutes. Discarded solution.
- Dehydrate in 95% alcohol and absolute alcohol, and clear in xylene, 2 changes each.
- Mount.
Shorten staining times when using 2 – 4 microns sections of renal biopsy specimens:
Biebrich Scarlet – Acid Fuchsin: 5 minutes
Phosphomolybdic Acid-Phosphotungstic Acid: 10-12 minutes
Aniline Blue Solution: 3 minutes
Stain Results:
Nuclei |
Black |
Cytoplasm, Keratin, Muscle Fibers |
Red |
Collagen, Mucin |
Blue |
NOTE: To ensure proper mordanting Bouin’s Fixative should be preheated to 58 – 60°C
References:
Sheehan and Hrapchak. Theory and Practice of Histotechnology. St. Lous. The Mosby Comp. 1980, p. 190
Product Information
Masson' Trichrome for Connective Tissue