TRICHROME STAIN FOR GMA OR MMA EMBEDDED SAMPLES
Submitted by Patsy Ruegg, HT(ASCP)QIHC    UCHSC Retired

Dry sections on 70dc hot plate for at least 20 minutes        30 min
(62dc oven)

Mordant slides in saturated picric acid

Wash in running tap
water                                                 10 min

Rinse in deionized water

Stain in extra strength Weigert's  - filtered
5 min

Rinse in running tap
water                                                 3-5 min

Stain in beibrich scarlet - acid fuchsin
30 min

Rinse quickly in deionized water

Stain in 2% lt. green in 2% acetic acid
10 min

Rinse in deionized water

3% acetic water for 4 to 5 min willenhance the blue

3% acetic in 95% alcohol, 2 changes, to remove stain from plastic media

Two changes of acetone

Clear in xylene and coverslip in permount

Results:  Nuclei-black; mineralized bone-green; osteoid-red/orange;
collagen-green or blue (depending on counterstain); keratin, muscle, and
intracellular fibers - red

Biebrich scarlet-acid fuchsin
  2% biebrich scarlet
  1% acid fuchsin
    Mix together:
      45 ml  bieb-scarlet  (Can be reused but filter before each use)
       5 ml  acid fuchsin
      Stir, then add 0.5 ml acetic acid

Extra Strength Weigert's
  Sol A  10 gm  Hematoxylin
            100 ml  95% alcohol
  Sol B   11.6 gm ferric chloride
            980 ml deionized or distilled water
              10 ml  25% HCL acid
Mix together and filter:
  Sol A    5 ML
  Sol B   25 ML
  Abs. alcohol  20 ml

Aniline Blue
  2.5 gm aniline blue
  2 ml acetic acid
  100 ml deionized or distilled water

2% Light Green
  2 gm light green
  98 ml distilled water
  2 ml acetic acid

(Procedure orginally developed by Mabel Myli, Mayo Clinic many moons
ago)