Masson's Trichrome Stain Kit

Masson's Trichrome Stain is used to demonstrate connective tissue elements, collagen and muscle fibers. After staining, collagen will be coloured in blue, muscle fibers, cytoplasm and keratin in red and the nuclei blue/black.

Use and sample preparation:
Formalin-fixed, paraffin and frozen samples can be used. After each step, rinsing with tap water and distilled water is required unless specified

Components

• Hematoxylin Weigert’s Reagent A
• Hematoxylin Weigert’s Reagent B
• Phosphotungstic Phosphomolibdic Acid Reagent
• Aniline Blue
• Glacial acetic acid 1%
• Bouin fixative reagent
• Biebrich scarlet

Short procedure

1. Place slide in Bouin’s reagent at 60ºC for 1 hour
2. Let the slides cool for 10 minutes
3. Mix equal parts of Weigert’s Reagent A and B
4. Place slide in Weigert’s Hematoxylin reagent freshly prepared for 5 minutes
5. Rinse slide with water
6. Apply Briebrich Scarlet for 15 minutes and do not rinse
7. Apply Phosphotungstic phosphomolybdic acid for 10-15 minutes. Do not rinse
8. Apply Aniline blue reagent for 5-10 minutes
9. Apply Glacial acetic acid 1% for 3-5 minutes
10. Dehydrate with ethanol, clear and coverslip

Stain characteristics:
Collagen can be visualized as blue, muscle fibers, cytoplasm and keratin in red and the nuclei blue/black