Skip to main content

Paquin & Goddard’s Trichrome for Elastic and Collagen

Paquin & Goddard's Trichrome

for Elastic and Collagen



  • Paquin & Goddard’s iron hematoxylin or equivalent
  • Solution A
    Eosin Y0.7g
    Orange G0.1g
    Phosphotungstic acid1g
    Distilled water1L
  • Solution B
    Phosphotungstic acid2g
    Distilled water1L
  • Solution C
    Acetic acid, glacial4mL
    Distilled water1L
  • Solution D
    Aniline blue0.4g
    Acetic acid, glacial10mL
    Distilled water990mL

Tissue Sample

5µ paraffin sections of Masson’s picric-chrome-formalin fixation was specified. Most trichrome stains benefit from picric acid or mercuric chloride fixation. The specified fixation may be compensated for by secondary fixation of sections in Bouin’s fluid.

Masson’s picric-chrome-formalin contains:

Picric acid, sat. aqu.187.5mL
Chromium aluminum sulphate7.5g
Formalin, Conc. 40%67.5mL
  1. Add the alum to the formalin and leave for 1 hour.
  2. Add the picric acid solution.
  3. Let stand for 24 hours, and filter.
  4. Return to tissue sample.


  1. Bring sections to water via xylene and ethanol.
  2. Stain with Paquin & Goddard’s iron hematoxylin.
  3. Wash with water.
  4. Place into solution A for 5 minutes.
  5. Place into solution B for 5 minutes.
  6. Rinse twice with solution C.
  7. Place into solution D for 5 minutes.
  8. Rinse twice with solution C.
  9. Place into solution B for 5 minutes.
  10. Place into solution C for 30 minutes.
  11. Rinse quickly with 95% ethanol.
  12. Dehydrate with iso-amyl alcohol.
  13. Clear with toluene and mount with a resinous medium.

Expected Results

  • Nuclei – black
  • Elastic – red
  • Cytoplasm – pink
  • Collagen – blue


  • If iso-amyl alcohol is not available, t-butanol is often suitable when the staining is likely to be removed with ethanol, although rapid dehydration with ethanol can often overcome this.
  • Although toluene was specified for clearing, xylene is likely suitable.

Safety Note

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


  1. Gray, Peter. (1954)
    The Microtomist’s Formulary and Guide.
    Originally published by: The Blakiston Co.
    Republished by: Robert E. Krieger Publishing Co.
    Paquin & Goddard, (1947)
    Bulletin of the International Association of Medical Museums and
    Journal of Technical Methods, v.27, pp.198