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Fullmer & Lillie’s Orcinol-New Fuchsin for Elastic Fibres

Fullmer & Lillie's Orcinol-New Fuchsin

for Elastic Fibres

5
steps
7
materials

Materials

    • Weigert’s iron hematoxylin or equivalent
    • Van Gieson’s picro-fuchsin
    • Orcinol-New Fuchsin solution
      MaterialAmount
      New fuchsin2g
      Orcinol4g
      Ferric chloride, 30% aqu.50mL
      Distilled water200mL
      Ethanol 95%100mL

      Preparation

      1. Add the dye and orcinol to the water in an oversize flask.
      2. Boil for 5 minutes.
      3. Add the ferric chloride.
      4. Boil for a further 5 minutes.
      5. Cool and filter.
      6. Dry the filter paper and beaker. Place the precipitate and filter paper back into the flask.
      7. Add the ethanol and heat carefully until the precipitate dissolves.
      8. Restore to 100 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 orcinol-new fuchsin solution for 15 minutes at 37°C.
  3. Wash with 3 changes of 70% ethanol for 5 minutes each.
  4. Counterstain with iron hematoxylin and van Gieson.
  5. Dehydrate with ethanol, clear with xylene and mount with a resinous medium.

Expected Results

  • Elastic fibres  –  deep violet
  • Collagen  –  red
  • Cytoplasm  –  yellow
  • Nuclei  –  black

Notes

  • Fullmer and Lillie specified 25mL USP IX Liquor Ferri Chloridi, or a solution of 15.5g ferric chloride (FeCl3.6H2O) made up to 25 mL with water, i.e. 25mL of a 62% solution. In either case, it should be fresh.

Safety Note

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

References

  1. Fullmer, H M and Lillie, R D, (1956).
    A selective stain for elastic tissue (orcinol-new fuchsin).,
    Stain Technologyl, v 31, page 27-29.
  2. Lillie, R D and Fullmer, H M, (1976).
    Histopathological technic and practical histochemistry, Ed. 4. pp. 714.
    McGraw-Hill, New York, USA.