Laboratory Protocols
Histological Staining Techniques
H.V.G. (Haematoxylin/van Gieson or Weigert/van Gieson) for connective tissue
Weigert 1904, van Gieson, 1889.
- Dewax sections, rinse in alcohol, rinse in water.
- Weigert's 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.
- Van Gieson's stain - 2 minutes.
- Rinse quickly in distilled water.
- Blot dry, rinse quickly in 100% alcohol, clear and mount. Any crystalline precipitate can usually be removed by taking back to distilled water.
Results:
Nuclei - grey/black.
Collagen - red.
Muscle, red blood cells, cytoplasm - yellow.
Weigert's haematoxylin
Solution A
- 1% haematoxylin in 100% alcohol.
Solution B
- 30% ferric chloride (anhydrous) - 4mls
- Concentrated HCl - 1ml
- Distilled water - 95mls
Add equal parts of A + B mix and use immediately.
Van Gieson's stain (Curtis' variation)
Saturated aqueous picric acid - 90mls
1% aqueous ponceau S - 10mls
Glacial acetic acid - 1.25mls