Duprès' Trichrome
for Muscle and Collagen
Materials
- Gallego’s carbol fuchsin or
Duprès’ magenta - Solution A
Material Amount Phosphomolybdic acid 1 g Distilled water 100 mL - Solution B
Material Amount Toluidine blue 0.25 g Orange G 4 g Oxalic acid 4 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. Most trichrome stains benefit from mercuric chloride fixation. Treatment with picric acid may be less satisfactory due to the use of the basic dye, toluidine blue.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei with either Gallego’s carbol fuchsin or Duprès’ magenta
- Wash with water.
- Place into solution A for 10 minute.
- Rinse with distilled water.
- Place into solution B for 1-5 minutes.
- Drain.
- Place into 95% ethanol until differentiated.
- Dehydrate with 100% ethanol.
- Clear with xylene and mount with a resinous medium
.
Expected Results
- Nuclei – blue black (Gallego) or red (Duprès)
- Erythrocytes – orange
- Cytoplasm – red
- Muscle – light green
- Bone – dark green
- Collagen – blue
Notes
- The celestine blue-hemalum sequence or Weigert’s iron hematoxylin may be suitable at step 2.
- Solution B contains both a basic and an acid dye.
- Other basic dyes may be substituted for toluidine blue:
- Methyl green 0.3 g
- Methylene blue 0.3 g
- Malachite green 0.2 g
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:
Duprès, (1935)
Journal number 11425, v. 46, pp. 77
Note: Gray identifies journals by a number from the world list of scientific periodicals, ed. 2, 1934. Unfortunately, the journal listed as number 11425 is not identified by name in the list he provides. In a second reference to the differentiator used in this technique he gives the journal number as 14425. This may be the intended reference:
Duprès, (1935)
Monitore zoologico italiano, v. 46, pp. 77
Florence, Italy.
