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Techniques. Protocols


This staining protocol combines a nuclear staining with Weigert hematoxylin and a picric acid/acid fuchsin solution, which stains collagen fibers of the connective tissue.


Whatever common fixative for general stainings is suitable for this protocol. Samples are embedded in paraffin and 8 µm thick sections are obtained and attached to gelatin coated slides.

1. 2x10 min in xylene

2. 2x10 min in 100º ethanol

3.- 10 min in 96º ethanol

4. 10 min in 80º ethanol

5. 10 min in 50º ethanol

6. 5 min in distilled H2O

7. 10 min in Weigert hematoxylin. This staining solution is composed of two separate solutions that are mixed immediately before use.

8. 5 min in tap water

9. 1 min in distilled H2O

10. 4 - 5 min in Van Gieson's pricrofuchsin
Van Gieson's picrofuchsin

Solution A. 1 % Acid fuchsin.
1 g acid fuchsin (C.I. 75290)
100 ml distilled H2O
Solution B. Saturated picric acid solution.
Working solution.
1 ml of solution A
45 ml of solution B

The working solution can be made just before the staining, although several weeks old solution can also be used by adjusting the staining time. Anyway, 0.25 ml of hydrochloric acid is added before use. Instead of just water, the following acidified water step is recommended to preserve a strong staining.

11. 2 x 30 s in acidified H2O
Acidified H2O:
0.5 ml of glacial acetic acid
100 ml of distilled H2O

12. 3 x 1 min in 100º ethanol

13. 2 x 5 min in xylene

14. Mounting media


Collagen: red - pink

Muscle: yellow-orange

cytoplasm: yellow - orange

Nuclei: blue -black



50º, 70º, 80º, 90º, 96º y 100º ethanol

Weigert hematoxylin

Acid fuchsin (C.I. 75290)

Picric acid

Galical acetic acid

Hydrochloric acid

Distilled H2O

Tap water

Mounting medium


Staining dishes

Slides racks


Van Gieson trichrome
8 µm thick section from paraffin embedded tissues stained with Van Gieson trichrome. Deep region of the dermis.
Home / Techniques / Protocols / Van Gieson trichrome