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Weigert’s Stain for Elastic Fibres

Protocol

Weigert's Stain

for Elastic Fibres

7
steps
8
materials
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Materials

  • Weigert’s iron hematoxylin or equivalent
  • Van Gieson’s picro-fuchsin
  • Weigert’s solution
    MaterialAmount
    Basic fuchsin2g
    Resorcin4g
    Ferric chloride, 30% aqu.25mL
    Ethanol 95%200mL
    Distilled water200mL
    Hydrochloric acid, conc.4mL

Preparation

  1. Add the dye and resorcin to the water in an oversize flask. Bring to the boil, and add the ferric chloride.
  2. Boil for 5 minutes.
  3. Cool and filter. Dry the filter paper and beaker. Place the precipitate and filter paper back into the flask.
  4. Add the ethanol and heat carefully until the precipitate dissolves.
  5. Add the hydrochloric acid.
  6. Restore to 200 mL with 95% ethanol.

Tissue Sample

5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.

Protocol

  1. Bring sections to water via xylene and ethanol.
  2. Place into Weigert’s solution for 20 minutes to 1 hour.
  3. Wash with 95% ethanol to remove excess solution.
  4. Differentiate with 1% acid alcohol.
  5. Wash in water.
  6. Counterstain with iron hematoxylin and van Gieson.
  7. 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

  • The procedure often benefits from a Mallory bleach.
  • It is usually recommended that the basic fuchsin should not be of the type that produces good Schiff’s reagent. That is, the basic fuchsin should contain more rosanilin than pararosanilin.
  • The ferric chloride solution should be freshly made.

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.

References

  1. Culling C.F.A., (1974)
    Handbook of histopathological and histochemical techniques, Ed. 3
    Butterworth, London, UK.
  2. Drury, R.A.B. and Wallington, E.A., (1980)
    Carleton’s histological technique, Ed. 5
    Oxford University Press, Oxford, UK.
  3. Humason, G. L., (1967)
    Animal tissue techniques, Ed. 2
    W. H. Freeman and Company, San Francisco, Ca, USA