Masson’s Trichrome Stain

Reagent Preparation

Prepare working Weigert’s Iron Hematoxylin solution (Catalog No. HT1079) by adding equal amounts of Part A and Part B. Prepare working Phosphotungstic-Phosphomolybdic Acid solution by mixing 1 volume of Phosphotungstic Acid solution with 1 volume of Phosphomolybdic Acid solution with two volumes of deionized water. Use once and discard.


  1. Deparaffinize tissue sections and hydrate to deionized water.
  2. Mordant in Bouin’s solution (Catalog No. HT10-1) at 56 °C for 15 minutes.
  3. Wash in running tap water until yellow color is removed.
  4. Stain in working Weigert’s Iron Hematoxylin solution for 5 minutes.
  5. Wash in running tap water for 5 minutes.
  6. Rinse in deionized water.
  7. Stain in Biebrich Scarlet Acid Fuchsin solution for 5 minutes.
  8. Rinse in deionized water.
  9. Place slides in working Phosphotungstic-Phosphomolybdic Acid solution for 5 minutes.
  10. Stain in Aniline Blue solution for 5 minutes.
  11. Place slides in 1% Acetic Acid solution for 2 minutes.
  12. Rinse slides, dehydrate through alcohol, clear in xylene and mount.


Nuclei – Black
Muscle fibers – Red
Collagen – Blue
Cytoplasm – Red

Masson's Trichrome Stain


Related Links