Lendrum, Slidders & Fraser's Trichrome
for Collagen and Other Tissues
Materials
Small Molecular Weight (MW) Solution
| Material | Amount | |
|---|---|---|
| Dye | 0.5 | g |
| Phosphotungstic acid | 2 | g |
| Ethanol, 95% | 100 | mL |
Use one of the dyes from the appropriate list below:
Small MW Dyes
| Dye | CI Name | Author’s Code |
|---|---|---|
| Martius yellow | Acid yellow 24 | AY24 |
| Orange G | Acid orange 10 | A010 |
| Azo-eosin | Acid red 4 | AR4 |
| Propalan red 3GX | Acid red 57 | AR57 |
| Acid fuchsin | Acid violet 19 | AV19 |
Large MW solution
| Material | Amount | |
|---|---|---|
| Dye | 0.5 | g |
| Acetic acid, glacial | 1 | mL |
| Distilled water | 99 | mL |
Use one of the dyes from the appropriate list below:
Large MW Dyes
| Dye | CI Name | Author’s Code |
|---|---|---|
| Sun yellow | Direct yellow 11 | DY11 |
| Sirius red F3B | Direct red 80 | DR80 |
| Benzo new blue GS | Direct blue 10 | DR10 |
| Durazol brilliant blue B | Direct blue 109 | DB109 |
Tissue Sample
3-5µ paraffin sections of formal sublimate fixed tissue is preferred. If formalin fixation is used, the sections may require secondary fixation with picro-mercuric-alcohol.
Protocol
- Bring sections to water via xylene and ethanol.
- Stain nuclei with an acid resistant nuclear stain.
- Rinse with 95% ethanol.
- Place into small MW solution for 3 minutes.
- Rinse with distilled water.
- Place in the large MW solution for 30 minutes to 2 hours.
- Rinse with water.
- Dehydrate with absolute ethanol.
- Clear with xylene and mount with a resinous medium.
Expected Results
The results vary depending on the particular combination of small and large molecular weight dyes chosen. As a general guide, the small molecular weight dye can be expected to stain erythrocytes, cytoplasm and muscle. The large molecular weight dye can be expected to stain collagen. The authors particularly recommended the following combinations:
- General use – AO10, DR80
- After PAS – AY24, DY11
- After silver – AR4, DY11
Notes
- This technique has not gained favour as it uses some difficult to obtain dyes.
- Picro-mercuric-alcohol is a saturated solution of both picric acid and mercuric chloride in absolute ethanol. It is usually applied to sections overnight after dewaxing and treating with ethanol, but before washing with water. If used, the iodine-thiosulphate sequence should be used to remove mercury pigment before staining.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Lendrum, A.C., Slidders, W. and Fraser, S., (1972),
Renal hyalin: A study of amyloidosis and diabetic fibrinous vasculosis with new staining methods.,
Journal of Clinical Pathology, v 25, page 373
