van Gieson's Stain
for Collagen
Materials
- Acid resistant nuclear stain: Weigert’s iron hematoxylin or Celestine blue hemalum
- Staining solution
Material Amount Saturated aqueous Picric acid 100 mL 1% aqueous Acid fuchsin 5-15 mL
Tissue Sample
Paraffin sections at 5µ are suitable. Many fixatives, including formalin, are satisfactory. Stains using acid dyes often benefit from picric acid or mercuric chloride fixation.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei with the acid resistant nuclear stain.
- Rinse well with tap water.
- Place into the staining solution for 2-5 minutes.
- Optionally, rinse quickly with distilled water.
- Dehydrate with ethanol.
- 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.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- 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.
