Goldner's Trichrome Method
EMS Catalog #26386
Fixation
10% Buffered Neutral Formalin or Bouin's Solution
Sections
Paraffin, frozen or celloidin
Microtomy
Cut paraffin sections at 6 micrometers.
Procedure
- Decerate slides in Xylene, 2 changes, 2 minutes each. Place in absolute alcohol, 2 changes, 2 minutes each; 95% alcohol, 2 changes, 2 minutes each; then rinse in distilled water.
- Mordant in Bouin's Fluid solution for 1 hour at 56°C, or overnight at room temperature, if formalin fixed.
- Cool and wash in running tap water until yellow color disappears.
- Rinse in distilled water.
- Place in Weigert's Hematoxylin for 10 minutes.
***To prepare Weigert's Solution, mix equal parts of:
Weigert's Iron Hematoxylin A
Weigert's Iron Hematoxylin B
*** stir vigorously.
- Wash in running tap water for 10 minutes.
- Rinse in distilled water.
- Stain in Ponceau Acid Fuchsin for 5 minutes.
- Wash in Acetic Acid, 1%.
- Place in Phosphomolybdic Acid-Orange G solution until collagen is decolorized.
- Rinse in Acetic Acid, 1% for 30 seconds.
- Stain in Light Green Stock Solution for 5 minutes.
- Rinse in Acetic Acid, 1%, for 5 minutes.
- Blot sections but do not allow to dry.
- Dip quickly in absolute alcohol.
- Dehydrate slides in 95% alcohol and absolute alcohol 3 changes each. Clear in Xylene, 3 changes.
- Mount coverglass with appropriate medium (refractive index 1.48-1.56).
Stain Results
Nuclear chromatin |
Brownish black |
Cytoplasm |
Bright red |
Erythrocytes |
Orange |
Muscle |
Red |
Collagen |
Green |
References
Luna, Lee G. Histopathologic Methods and Color Atlas of Special Stains and Tissue Artifacts
Johnson Printers, Downers Grove , IL p 151-152, c 1992.
Product Information
Bouin’s Fluid
Weigert’s Iron Hematoxylin A
Weigert’s Iron Hematoxylin B
Ponceau Acid Fuchsin
Acetic Acid 1%
Phosphomolybdic Acid Orange G Solution
Light Green Stock 0.2%
Goldner's Trichrome Method