van Gieson
for Collagen

Solutions
An acid resistant nuclear stain, such as Weigert's iron hematoxylin or the Celestine blue hemalum sequence

Staining solution
Saturated aqueous Picric acid	100	mL
1% aqueous Acid fuchsin	5-15	mL

Fixation
Many fixatives, including formalin, are satisfactory.
Stains using acid dyes often benefit from picric acid or mercuric chloride fixation.

Tissue sample
Paraffin sections at 5µ are suitable.

Method

1. Bring sections to water via xylene and ethanol.
2. Stain nuclei with the acid resistant nuclear stain.
3. Rinse well with tap water.
4. Place into the staining solution for 2-5 minutes.
5. Optionally, rinse quickly with distilled water.
6. Dehydrate with ethanol.
7. Clear with xylene and mount with a resinous medium.

Expected results

• Collagen - red
• Cytoplasm - yellow
• Nuclei - black

Notes
This method is often used to counterstain other primary staining methods. In that case the nuclear stain may not be necessary and should be ommitted.

Reference
Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.

Drury, R.A.B. and Wallington, E.A., (1980)
Carleton's histological technique Ed. 5
Oxford University Press, Oxford, UK.

Culling C.F.A., (1974)
Handbook of histopathological and histochemical techniques Ed. 3
Butterworth, London, UK.