Histological Staining Techniques

Goldner's Trichrome for Osteoid
Goldner
  • Dewax sections, rinse in alcohol, rinse in water.
  • Stain nuclei with Weigert's iron haematoxylin - 20 minutes.
  • Wash in tap water - 1 minute.
  • Differentiate in acid alcohol (1% hydrochloric acid in 70% alcohol) - not more than 5 seconds.
  • Wash in tap water - 5 minutes.
  • Rinse in distilled water.
  • Stain in solution A for 5 minutes.
  • Rinse with 1% aqueous acetic acid.
  • Treat with solution B until collagen is decolourised. (Approx. 3 minutes.)
  • Rinse with 1% aqueous acetic acid.
  • Stain in solution C for 5 minutes.
  • Treat with 1% aqueous acetic acid for 5 minutes.
  • Blot dry, rinse quickly in 100% alcohol, clear and mount.

Results: 


Mineralised bone - green.
Osteoid - orange/red.
Nuclei blue - grey.
Cartilage - purple.

Solution A
Ponceau 2R - 0.075g
Acid fuchsin - 0.025g
Azophloxine - 0.01g
Acetic acid - 0.2ml
Distilled water - 100ml

Solution B
Orange G - 2g
Phosphomolybdic acid (dodeca-molybdophosphoric acid)- 4g
Distilled water - 100ml
Leave to stand for a couple of days for any precipitate to settle out and then filter before using.

Solution C
Light green - 0.2g
Acetic acid - 0.2ml
Distilled water - 100ml

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