Klatskin's Modification of Masson's Trichrome
for Liver
Materials
- Harris’ alum hematoxylin.
- Solution A
Material Amount Picric acid to saturation Ethanol, 95% 100 mL - Solution B
Material Amount Xylidine ponceau 1 g Distilled water 99 mL Glacial acetic acid 1 mL - Solution C
Material Amount Phosphomolybdic acid 1 g Distilled water 100 mL - Solution D
Material Amount Aniline blue to saturation Glacial acetic acid 2.5 mL Distilled water 97.5 mL - Solution E
Material Amount Acetic acid, glacial 1 mL Distilled water 99 mL
Tissue Sample
3-5 micron paraffin sections of formalin fixed liver.
Protocol
- Bring sections to water via xylene and ethanol
- Stain with Harris’ hemalum for 10 minutes.
- Rinse with 95% ethanol.
- Place in solution A for 10 minutes.
- Wash with water for 10 minutes.
- Place in solution B for 5 minutes.
- Place in solution C for 5 minutes.
- Place in solution D for 2 minutes.
- Replace in solution C for 5 minutes.
- Place in solution E for 5 minutes.
- Place in 70% ethanol for 2 minutes.
- Dehydrate with ethanol, clear and mount with a resinous medium.
Expected Results
- Cytoplasm – red
- Erythrocytes – red
- Muscle – red
- Collagen – blue
- Nuclei – brown
Notes
- Aniline blue, is a mixture of two dyes, methyl blue, and water blue. Either may usually be substituted satisfactorily.
- Although recommended for liver sections, this modification may also be used as a general replaceement for Masson’s method.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- 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