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MODIFIED MASSONS TRICHROME STAIN FOR CARDIAC TISSUES
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FIXATION: 10% buffered neutral formalin
TECHNIQUE: Paraffin sections cut at 5 μm.
SOLUTIONS:
Modified Weigerts Iron Hematoxylin
Solution A
Hematoxylin, C.I. 75290 -------------------------------------------------------- 2.0 gm
90% ethyl alcohol ---------------------------------------------------------------- 100.0 ml
Solution B
Ferric chloride, FeCl3, 6H20, 62% --------------------------------------------- 4.0 ml
Distilled water -------------------------------------------------------------------- 95.0 ml
Hydrochloric acid, concentrated ----------------------------------------------- 1.0 ml
Working Modified Weigerts Iron Hematoxylin
Equal parts of Solution A and Solution B
Alcoholic Picric Acid Solution
Picric acid, saturated aqueous ----------------------------------------------------- 10.0 ml
Alcohol, 95% --------------------------------------------------------------------- 40.0 ml
1% Ponceau S Solution
Ponceau S, C.I. 27195 ----------------------------------------------------------- 0.5 gm
Distilled water -------------------------------------------------------------------- 50.0 ml
Acetic acid ------------------------------------------------------------------------ 0.5 ml
1% Phosphomolybdic Acid Solution
Phosphomolybdic acid ---------------------------------------------------------- - 1.0 gm
Distilled water -------------------------------------------------------------------- 100.0 ml
Aniline Blue Solution
Aniline blue, C.I. 42755 -------------------------------------------------------- -- 1.0 gm
Distilled water -------------------------------------------------------------------- 44.0 ml
Acetic acid ------------------------------------------------------------------------ 0.9 ml
1% Acetic Acid Solution
Acetic acid ------------------------------------------------------------------------ 1.0 ml
Distilled water -------------------------------------------------------------------- 99.0 ml
9. Place in ponceau S solution in a glass Coplin jar. Place in a microwave oven and microwave at power level 1 (60W) for 3 minutes. Agitate slides for about 15 seconds and allow to set in the warm solution for 2 minutes.
10. Rinse with three changes of distilled water.
11. Place in phosphomolybdic acid solution for 5 minutes.
12. Rinse quickly with two changes of distilled water.
13. Place in aniline blue solution for 2 minute.
14. Place in 1% acetic acid solution for 1 minute.
15. Rinse quickly with two changes of distilled water.
16. Dehydrate in graded alcohols.
17. Clear in three or four changes of xylene.
18. Mount with synthetic resin.
Nuclei -------------------------------------------------------------- blue-black
Intercellular fibers --------------------------------------------------- red
Collagen ------------------------------------------------------------- blue
Cytoplasm ----------------------------------------------------------- red
Mucin --------------------------------------------------------------- blue
Keratin -------------------------------------------------------------- red
Argentaffin granules ------------------------------------------------- black or red
Neuroglia fibers ----------------------------------------------------- red
Masson, P.J.: Some histological methods. Trichrome stainings and their preliminary technique. J. Tech. Methods 12:75-90, 1929.
Clark, G.: Staining Procedures, 4th edition, Baltimore, Williams and Wilkins, 1981, pp. 118-120.
Image Examples:
Diagnostic Application: