Special Stains - Van Gieson

Van gieson is a Connective tissue stain to differentiate collagen by using dyes of different molecular size. This special stain is used for staining muscle and collagen fibres.  This uses two dyes, acid fuschin and picric acid.  The nuclear stain used is haematoxylin after being through celestein blue.  This produces a darker black colour than haematoxylin alone.  The acid dyes used stains collagen red and muscle yellow in the tissue.

Method

1.    Take sections to water.

2.     Stain using Celestin blue / haematoxylin sequence.

3.     Wash in water.

4.     Differentiate in 1% acid alcohol if necessary.

5.     Wash in water.  Blue Lithium Carbonate

6.     Stain in Van Gieson's solution for 3 minutes.

7.     Blot

8.     Dehydrate, clear and mount.

Results

Nuclei	Blue/Black
Collagen	Red
Cytoplasm, Muscle, RBCs	Yellow
Some bile pigments	Green

The results can be seen below on the liver biopsy, the collagen is shown up clearly in red while the background is stained yellow providing a much better contrast.

Photo - x10 Van gieson