Pasini's Trichrome
for Elastic and Collagen
Materials
- Ehrlich’s alum hematoxylin
- Solution A
Material Amount Phosphotungstic acid 2 g Distilled water 100 mL - Solution B – Var I
Material Amount Eosin B 0.7 g Acid fuchsin, sat. aqu. 4 mL Unna’s elastic stain 35 mL Glycerol 40 mL Ethanol, 50% 35 mL - Solution B – Var II
Material Amount Eosin B, 2% in 50% ethanol 30 mL Acid fuchsin, sat. aqu. 4 mL Unna’s elastic stain 30 mL Glycerol 15 mL
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 an alum hematoxylin nuclear stain.
- Place into solution A for 10 minute.
- Rinse quickly with distilled water.
- Place into solution B for 15-20 minutes.
- Rinse quickly with 70% ethanol.
- Dehydrate and differentiate with 100% ethanol for a few seconds.
- Clear with xylene and mount with a resinous medium.
Expected Results
- Nuclei – blue
- Elastic fibres – purple
- Erythrocytes – orange
- Collagen – blue
Notes
- Solution B var I was given by Gray, var II was given by Gatenby & Beams.
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:
Gatenby, J. B. & Cowdry, E. V., (1928)
Bolles Lee’s Microtomist’s Vade Mecum, pp. 280
Blakiston, Philadelphia - Bolles Lee, A.. Edited by Gatenby, J.B. and Beams, H.W., (1950)
The Microtomist’s Vade-Mecum. 11 ed., pp. 485
Churchill, London, UK.
