Lewis and Miller's Trichrome
for Pituitary cells

This is a modification of Kricheski's method

Solutions

Solution A
Acid fuchsin	0.25	g
Distilled water	100	mL

Solution B
Methyl blue, 1% aqueous	30	mL
Orange G, 1% aqueous	30	mL
Phosphomolybdic acid, 1% aqueous	30	mL

Tissue sample
5µ paraffin sections of neutral buffered formalin fixed tissue are likely suitable. Other fixatives are also 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 30 minute.
3. Rinse well with distilled water.
4. Place into solution B for 24 hours.
5. Dip 2 or 3 times in 70% ethanol.
6. Dehydrate and differentiate with ethanol for 1-3 minutes.
7. Clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  red
• Acidophils  –  orange red
• Basophils  –  blue
• Chromophobes  –  Pale grey

Notes

1. Although not specified, an acid resistant nuclear stain such as Weigert's iron hematoxylin could be inserted prior to staining with solution A.

 

Reference

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
  Citing:–
    Kricheski, (1931)
    Stain technology, v.6, pp.97
  And:–
    Lewis and Miller, (1938)
    Stain technology, v.14, pp.111