Lee-Brown's Trichrome
for Muscle and Collagen

This is a modification of Mallory's trichrome

Solutions

Solution A
Acid fuchsin 1 g
Distilled water 100 mL
Directions
Solution B
Phosphomolybdic acid 1 g
Distilled water 100 mL
Directions
Solution C
Orange G 2 g
Methyl blue 0.5 g
Oxalic acid 2 g
Distilled water 100 mL

Tissue sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory. Most trichrome stains benefit from picric acid or mercuric chloride fixation. Formalin fixed tissues may benefit from secondary fixation of sections in Bouin's fluid.

Method

  1. Bring sections to water via xylene and ethanol.
  2. Place into solution A for 2 minute.
  3. Rinse with distilled water.
  4. Place into solution C for 15 minutes.
  5. Rinse quickly with distilled water.
  6. Place into solution B for 2 minutes.
  7. Wash well with distilled water.
  8. Dehydrate and differentiate with ethanol.
  9. Clear with xylene and mount with a resinous medium.

Expected results

Notes

  1. This modification reverses the staining sequence of Mallory's original method, and applies solution C before solution B.
  2. Although an acid resistant nuclear stain is not specified, Weigert's iron hematoxylin or an equivalent could be done before step 2. The nuclei would then be black.

 

Reference

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
  Citing:–
    Mallory, (1901)
    Journal of Experimental Medicine, v.5, pp.15
  And:–
    Lee-Brown, (1929)
    Journal of Urology, v.21, pp.259

 


 

 

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