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Lendrum, Slidders & Fraser’s Trichrome for Collagen and Other Tissues

Lendrum, Slidders & Fraser's Trichrome

for Collagen and Other Tissues



Small Molecular Weight (MW) Solution

Phosphotungstic acid2g
Ethanol, 95%100mL

Use one of the dyes from the appropriate list below:

Small MW Dyes

DyeCI NameAuthor’s Code
Martius yellowAcid yellow 24AY24
Orange GAcid orange 10A010
Azo-eosinAcid red 4AR4
Propalan red 3GXAcid red 57AR57
Acid fuchsinAcid violet 19AV19

Large MW solution

Acetic acid, glacial1mL
Distilled water99mL

Use one of the dyes from the appropriate list below:

Large MW Dyes

DyeCI NameAuthor’s Code
Sun yellowDirect yellow 11DY11
Sirius red F3BDirect red 80DR80
Benzo new blue GSDirect blue 10DR10
Durazol brilliant blue BDirect blue 109DB109

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.


  1. Bring sections to water via xylene and ethanol.
  2. Stain nuclei with an acid resistant nuclear stain.
  3. Rinse with 95% ethanol.
  4. Place into small MW solution for 3 minutes.
  5. Rinse with distilled water.
  6. Place in the large MW solution for 30 minutes to 2 hours.
  7. Rinse with water.
  8. Dehydrate with absolute ethanol.
  9. 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


  • 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.


  1. 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