Gomori Trichrome Stain for Paraffin Embedded Tissue

1. Deparaffinize in xylene 2 X 5 min

2. Rehydrate in the following series or alcohols:

100% - 2 X 3 min

95% - 2 X 3 min

70% - 1 X 3 min

3. Rinse in water

4. For ISH or formalin fixed tissues only: place in Bouin’s solution in oven at 56ºC for 1 hr.

5. Wash well in running water.

6. Stain nuclei with hematoxylin for 10 min.

7. Wash in running water for 10 min.

8. Trichrome stain for 5-20 min.

9. Dip in 0.5% acetic acid at intervals. If the stain is too dark, decolorize in 1% acetic acid plus 0.7% phosphotungstic acid.

10. Dehydrate in graded ethanol solutions – 95% 2 X 3 min, 100% 2 X 3 min

11. Xylene 2 X 3 min.

12. Mount coverslips with non-aqueous mounting medium.

Bouin’s Solution

Picric Acid, saturated aqueous solution 75ml

37% Formalin 25ml

Glacial Acetic Acid 5ml

Trichrome Stain

Chromotrope 2R 0.6g 1.8g

Fast Green FCF 0.3g 0.9g

Acetic Acid 1ml 3ml

Phosphotungstic Acid 0.8g 2.4g

Distilled Water 100ml 300ml

Keep stain in a dark container in the refrigerator.