Goldner's Trichrome
for Muscle and Collagen

Solutions

Weigert's iron hematoxylin or equivalent

Solution A		var 1			var 2			var 3
Ponceau 2R		0.7	g		0.75	g		–
Acid fuchsin		0.3	g		0.25	g		–
Azophloxine		–			0.1	g		5	g
Acetic acid, glacial		2	mL		2	mL		2	mL
Distilled water		1	L		1	L		1	L

Solution B
Acetic acid, glacial		10	mL
Distilled water		1	L

Solution C
Orange G		20	g
Phosphomolybdic acid		40	g
Distilled water		1	L

Solution D
Light green SF		2	g
Acetic acid, glacial		2	mL
Distilled water		1	L

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.

Method

1. Bring sections to water via xylene and ethanol.
2. Stain nuclei with Weigert's iron hematoxylin, or equivalent.
3. Place into one of the variants of solution A for 5 minutes.
4. Rinse with solution B.
5. Place into solution C until collagen is decolourised.
6. Rinse with solution B.
7. Place into solution D for 5 minutes.
8. Place into solution B for 5 minutes.
9. Blot.
10. Dehydrate rapidly with absolute ethanol.
11. Clear with xylene and mount with a resinous medium.

Expected results

• Nuclei  –  black
• Erythrocytes  –  orange
• Cytoplasm  –  red
• Collagen  –  green

 

Reference
Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.