Dahlia-Iron-Resorcin
for Elastic Fibres

Solutions

Weigert's iron hematoxylin or equivalent
Van Gieson's picro-fuchsin
Iron-resorcin-dahlia solution
Preparing the sludge
Dahlia 2 g   Bring 200 mL of 0.5% hydrochloric acid in an oversize flask to the boiling point, remove from the heat and carefully add the dahlia. Return to the boil, and add the resorcin and ferric nitrate. Continue boiling for 3 minutes, then cool and filter. Discard the filtrate.
Resorcin 4 g
Ferric nitrate, 30% aqu. 50 mL
0.5% aqu. hydrochloric acid 200 mL
 
Solvent
Methanol 180 mL   Dry the filter paper and beaker, then place the precipitate and filter paper back into the flask. Add the methanol, acetone and hydrochloric acid. Heat carefully on a hot plate until the precipitate dissolves.
Acetone 20 mL
Hydrochloric acid, conc. 4 mL

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

Method

  1. Bring sections to water via xylene and ethanol.
  2. Do a Mallory bleach.
  3. Place into the staining solution for 30 minutes or longer.
  4. Wash with 95% ethanol to remove excess solution.
  5. Differentiate with 1% acid alcohol if necessary.
  6. Wash in water.
  7. Counterstain with iron hematoxylin and van Gieson.
  8. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected results

Notes

  1. Between the 1st and 2nd editions of their book, Bancroft and Stevens changed the solvent for the precipitate to the following:
    Solvent
    2-methoxyethanol 100 mL   Dry the filter paper and beaker, then place the precipitate and filter paper back into the flask. Add the mixture. Heat carefully on a hot plate until the precipitate dissolves.
    Distilled water 100 mL
    Hydrochloric acid, conc. 4 mL
  2. 2-methoxyethanol is also known as ethylene glycol monomethyl ether.
  3. The authors suggest the following dyes as suitable substitutes for dahlia:
    Methyl violet, Ethyl violet, Victoria blue, Thionin,
  4. Note that this method uses ferric nitrate instead of ferric chloride.

 

Reference
Bancroft, J. D. and Stevens, A.,
Theory and Practice of Histological Techniques, Ed. 1 (1977),
Churchill Livingstone, London, UK.

Bancroft, J. D. and Stevens, A.,
Theory and Practice of Histological Techniques, Ed. 2 (1982),
Churchill Livingstone, London, UK.

 


 

 

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