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Electron Microscopy Sciences

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 (#26367-04) 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

Online Ordering

Masson' Trichrome for Connective Tissue is available online from the EMS Catalog. For ordering or product information, click here.