Ladewig's Trichrome
for Muscle and Collagen

Solutions

Weigert's iron hematoxylin or equivalent

Solution A
Phosphotungstic acid	1	g
Distilled water	100	mL

Solution B
Methyl blue	0.5	g
Orange G	2	g
Acid fuchsin	1	g
Oxalic acid	0.6	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. Many trichrome stains benefit from Bouin or mercuric chloride fixation. Sections of formalin fixed tiddue may benefit from secondary fixation in Bouin's fluid.

Method

1. Bring sections to water via xylene and ethanol.
2. Stain nuclei with Weigert's iron hematoxylin for 3-5 minutes.
3. Rinse with water.
4. Place into solution A for 2 minute.
5. Rinse with distilled water.
6. Place into solution B for 4 minutes.
7. Rinse quickly with distilled water.
8. Dehydrate with ethanol.
9. Clear with xylene and mount with a resinous medium.

Expected results

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

Notes

1. Times may need to be adjusted, particularly if fixatives other than formalin have been used.

 

Reference

Gray, Peter. (1954)
The Microtomist's Formulary and Guide.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.
  Citing:–
    Ladewig, (1938)
    Zeitschrift für wissenschaftliche Mikroskopie und für mikroskopische Technik,
    v.55, pp.215

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