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van Gieson Staining Protocol
NovaUltra Special Stain Kits
Principle
Van Gieson is used to differentiate between collagen and smooth muscle in tumours and to demonstrate the increase of collagen in diseases.
When using combined solutions of picric acid and acid fuchsin, the small molecules of picric acid penetrate all of the tissues rapidly, but are only firmly retained in the close textured, red blood cells and muscle. The larger molecules of ponceau S displaces picric acid molecules from collagen fibres, which have larger pores, and allow the larger molecules to enter.
Method
1 Bring sections to distilled water.
2 Stain nuclei with Celestin Blue 5 mins
3 Rinse in distilled water
4 Stain in haematoxylin 5 mins
5 Wash well in running tap water 5 mins
6 Flood with Curtis stain 5 mins
7 Blot.
8 Dehydrate rapidly in alcohols, clear and mount.
Results
Nuclei ....................................................Blue
Collagen .................................................Bright red
Cytoplasm, muscle, fibrin and red blood cells ......Yellow
Reagent Formulae
1. Celestin Blue
5% ammonium ferric sulphate (iron alum) 100ml
Celestin Blue (CI 51050) 0.5 g
Add the celestin blue to the ammonium ferric sulphate and boil for 3 minutes.
Filter when cool. Store refrigerated.
2. Curtis
Stain:
saturated aqueous picric acid 90.0 ml
1% ponceau S 10.0 ml
glacial acetic acid 10.0 ml
3. 1% Ponceau S
Ponceau S (CI 27195)
1.0 gm
distilled water
100.0 ml
References
Bancroft, J.D. and Cook, H.C. 1984, Manual of Histological Techniques. Churchill Livingstone.
Curtis F. Methode de colorationelective du tissue conjonctif. C R soc Biol 1905;58:1038-40