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Lillie’s Allochrome

Protocol

Lillie's Allochrome

8
steps
5
materials

Materials

  • 1% Aqueous periodic acid
  • Schiff’s reagent
  • Weigert’s iron hematoxylin
  • Picro-methyl blue
    MaterialAmount
    Saturated aqueous picric acid100mL
    1% aqueous methyl blue0.4mL

Tissue Sample

5 µ paraffin sections. Most fixatives are satisfactory, including 10% NBF, but those generally considered preferable for trichrome stains will be advantageous.

Protocol

  1. Bring sections to water via xylene and ethanol.
  2. Do a PAS stain, but do not counterstain.
  3. Stain nuclei with Weigert’s iron hematoxylin or equivalent.
  4. Wash well in tap water, rinse with distilled water.
  5. Place into picro-methyl blue for 6 minutes.
  6. Rinse well with 95% ethanol.
  7. Dehydrate with absolute ethanol.
  8. Clear with xylene and mount with a resinous medium.

Expected Results

  • Nuclei – Brown-black
  • Collagen & reticulin – blue
  • Cytoplasm & muscle – yellow
  • PAS positive structures – red

Safety Note

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

References

  1. Histological demonstration techniques, (1974)
    Cook, H C.
    Butterworths, London, England