Klatskin's Modification
of Masson's trichrome for Liver

Solutions

Harris' alum hematoxylin.

Solution A
Picric acid	to	saturation
Ethanol, 95%	100	mL

Solution B
Xylidine ponceau	1	g
Distilled water	99	mL
Glacial acetic acid	1	mL

Solution C
Phosphomolybdic acid	1	g
Distilled water	100	mL

Solution D
Aniline blue	to	saturation
Glacial acetic acid	2.5	mL
Distilled water	97.5	mL

Acetic acid
Acetic acid, glacial	1	mL
Distilled water	99	mL

Tissue sample
3-5 micron paraffin sections of formalin fixed liver.

Method

1. Bring sections to water via xylene and ethanol
2. Stain with Harris' hemalum for 10 minutes.
3. Rinse with 95% ethanol.
4. Place in solution A for 10 minutes.
5. Wash with water for 10 minutes.
6. Place in solution B for 5 minutes.
7. Place in solution C for 5 minutes.
8. Place in solution D for 2 minutes.
9. Replace in solution C for 5 minutes.
10. Place in solution E for 5 minutes.
11. Place in 70% ethanol for 2 minutes.
12. Dehydrate with ethanol, clear and mount with a resinous medium.

Expected results

• Cytoplasm  –  red
• Erythrocytes  –  red
• Muscle  –  red
• Collagen  –  blue
• Nuclei  –   brown

Notes

1. Aniline blue, is a mixture of two dyes, methyl blue, and water blue. Either may usually be substituted satisfactorily.
2. Although recommended for liver sections, this modification may also be used as a general replaceement for Masson's method.

Reference
Petersen, K.F., West, A.B., Reuben, A., Rothman, D. and Shulman, G.I. (1996)
Noninvasive Assessment of Hepatic Triglyceride Content in Humans With ¹³C Nuclear Magnetic Resonance Spectroscopy.
Hepatology, V. 24, p 114-117
This reference is available on the internet.

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