EMS Technical Data Sheets

Masson's Trichrome for Connective Tissue

EMS Catalog #: 26367-Series    


Bouin’s (#15990-01) or 10% Buffered Formalin (#15741-01)


Paraffin at 6µm

Staining Procedures:

  1. Deparaffinize and hydrate to distilled water.
  2. Mordant in Bouin’s Fixative (#26367-01) for 1 hour at 56°C or overnight at room temperature.
  3. Cool and wash in running water until yellow color disappears.
  4. Rinse in distilled water.
  5. Stain in Weigert’s Iron Hematoxylin Working* solution for 5 minutes. Wash in running water for 10 minutes.
    1. To prepare working solution, mix equal parts of Weigert’s Iron Hematoxylin A (#26367-02) and Weigert’s Iron Hematoxylin B (#26367-03).
  6. Rinse in distilled water.
  7. Place in Biebrich Scarlet-Acid Fuchsin (#26033-25) for 15 minutes.
  8. Rinse in distilled water.
  9. Place in Phosphomolybdic Acid-Phosphotungstic Acid (#26367-05) for 15 minutes.
  10. Stain in Aniline Blue Solution (#26367-06) for 10 to 20 minutes.
  11. Rinse in distilled water.
  12. Differentiate in Acetic Acid, 1% (#26367-07) for 3 to 5 minutes. Discarded solution.
  13. Dehydrate in 95% alcohol and absolute alcohol, and clear in xylene, 2 changes each.
  14. 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


Sheehan and Hrapchak. Theory and Practice of Histotechnology. St. Lous. The Mosby Comp. 1980, p. 190

Product Information

Masson' Trichrome for Connective Tissue