Hart's Stain
for Elastic Fibres
Materials
- Weigert’s iron hematoxylin or equivalent
- Van Gieson’s picro-fuchsin
- Hart’s solution
Material Amount Weigert’s iron resorcin fuchsin 1 volume 1% hydrochloric acid in 70% ethanol 9 volumes
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Place into Hart’s solution overnight.
- Wash with 95% ethanol to remove excess solution.
- Differentiate with 1% acid alcohol.
- Wash with tap water.
- Counterstain with iron hematoxylin and van Gieson.
- Dehydrate with ethanol, clear with xylene and mount with a resinous medium.
Expected Results
- Elastic fibres – blue black
- Collagen – red
- Cytoplasm – yellow
- Nuclei – blue
Notes
- This modification should be used if poor staining is obtained using Weigert’s method.
- The optimum dilution may be more or less than given.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Culling C.F.A., (1974)
Handbook of histopathological and histochemical techniques Ed. 3
Butterworth, London, UK.
