Duprès' trichrome
for Muscle and Collagen

Solutions

Gallego's carbol fuchsin or Duprès' magenta

Solution A
Phosphomolybdic acid	1	g
Distilled water	100	mL

Solution B
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.

Method

1. Bring sections to water via xylene and ethanol.
2. Stain nuclei with either Gallego's carbol fuchsin or Duprès' magenta
3. Wash with water.
4. Place into solution A for 10 minute.
5. Rinse with distilled water.
6. Place into solution B for 1-5 minutes.
7. Drain.
8. Place into 95% ethanol until differentiated.
9. Dehydrate with 100% ethanol.
10. 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

1. The celestine blue-hemalum sequence or Weigert's iron hematoxylin may be suitable at step 2.
2. Solution B contains both a basic and an acid dye.
3. Other basic dyes may be substituted for toluidine blue:
   Methyl green 0.3 g
   Methylene blue 0.3 g
   Malachite green 0.2 g

 

Reference

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)
    Monitore zoologico italiano, vol.46, pp.46 & 77