MASSON TRICHROME

MASSON TRICHROME

The trichrome are SO called for their ability to differentiate tissue element into 3 colors, Collagen which is relatively permeable stains blue with aniline blue and muscle his far less permeable stains red with acid fuchsin.

Solutions

1.   Weigart’s Iron Haematoxylin

Solution A

Haematoxylin            -           1gm
Absolute Alcohol       -           100ml
Solution B

30% Aqueous ferric chloride                               -           4ml
Concentrated hydrochloric acid              -         lml
 Distilled water                                         -         5ml
Mix equal quantities of A and B before use. The Solution should be dark black in color .

2.   Biebrich Scarlet ( Red color )

Biebrich Scarlet ( Aqueous ) 1%          -        90ml
Acid Fuchsin ( Aqueous ) 1%              -        10ml
Glacial Acetic Acid                              -        01ml




3.   Phosphotungstic Phosphomolybdic acid

            Phosphomolybdic acid                  -        5gm
          Phosphotungstic acid                         -         5gm
            Distilled Water                                   -         1400ml
            This solution is used to differentiate trichrome stains.

4.   Aniline Blue

Aniline Blue                  -           2.5gm
Glacial Acetic Acid       -            2ml
Distilled Water               -           100ml

5.   1% Acetic Acid



















Procedure

1.      Deparaffinise the section in xylene for 5-10 mts
2.      The section should be fixed in bouin's fluid for 1 hr. Bring the sections through
descending series of alcohol.
3.    Stain with iron Haematoxylin for 5-10 mnts .
4.    Wash in tap water
5.    Differentiate with 1% Acid Alcohol.
6.    Keep it in running tap water for 5-10 mnts
7.    Stain with Biebrich scarlet for 5 – 10 mnts
8.    Wash it in distilled water
9.    Treat with Phosphotungstic phosphomolybdic acid for 10 mnts
10. Wash in water
11. Stain with Aniline blue for 5 mnts
12. Drain the solution and differentiate in 1% acid alcohol
13. Wash in water , Dehydrate , Clear and Mount


Result
Nuclei                         -           Black
Collagen                     -           Blue
Muscle                        -           Red







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