Protocol
Pollak's Trichrome
for Muscle and Collagen
Materials
- Weigert’s iron hematoxylin or equivalent
- Solution A
Material Amount Orange G 0.25 g Ponceau 2R 0.33 g Acid fuchsin 0.17 g Light green SF 0.15 g Phosphotungstic acid 0.5 g Phosphomolybdic acid 0.5 g Acetic acid, glacial 1 mL Ethanol, 95% 50 mL Distilled water 50 mL - Solution B
Material Amount Acetic acid, glacial 0.2 mL Distilled water 100 mL
Preparation of Solution A
- Mix the water, ethanol and acetic acid together.
- Divide into four, and dissolve as follows:
- Phosphomolybdic acid
- Phosphotungstic acid and orange G
- Light green SF
- Ponceau 2R and acid fuchsin
- Combine all solutions and filter.
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. Formalin fixed tissues may benefit from secondary fixation of sections in Bouin’s fluid.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei with Weigert’s iron hematoxylin or equivalent.
- Place into solution A for 3-7 minute.
- Place into solution B until differentiated.
- Dehydrate with ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – black
- Erythrocytes – yellow
- Cytoplasm – red
- Muscle – red
- Collagen – blue
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Gray, Peter. (1954)
The Microtomist’s Formulary and Guide.
Originally published by: The Blakiston Co.
Republished by: Robert E. Krieger Publishing Co.
Citing:
Pollack, (1944)
Archives of pathology and laboratory medicine, v. 37, pp. 294
Chicago, USA
