Van Gieson's Stain is a mixture of Picric Acid

Picric acid

Picric acid is the chemical compound formally called 2,4,6-trinitrophenol . This yellow crystalline solid is one of the most acidic phenols. Like other highly nitrated compounds such as TNT, picric acid is an explosive...

 and Acid Fuchsin

Acid fuchsin

Acid fuchsin is an acidic dye that has wide usage in histology. Acid Fuchsin is one of the dyes used in Masson's Trichrome Stain. This method is commonly used to stain tissue sections in the Histology Laboratory in order to distinguish muscle from collagen...

. It is the simplest method of differential staining of Collagen

Collagen

Collagen is a group of naturally occurring proteins found in animals, especially in the flesh and connective tissues of mammals. It is the main component of connective tissue, and is the most abundant protein in mammals, making up about 25% to 35% of the whole-body protein content...

 and other Connective Tissue. It was introduced to histology by American neuropsychiatrist and pathologist Ira Van Gieson

Ira Van Gieson

Ira Thompson Van Gieson was an American neurologist, psychiatrist, bacteriologist and neuropathologist....

.

Formula

• 100 mL of Saturated Aqueous solution of Picric Acid added to 5 mL of 1% Aqueous solution of Acid Fuchsin.

The solution weakens after long standing and may be strengthened by adding few drops of fresh Acid Fuchsin.

Staining Connective Tissue with Van Gieson's

Recommended Fixative

• Mercuric Chloride fixative, although Formol-Saline also gives good results.

Procedure

• 1) Tissue sections to Ethyl Alcohol
• 2) Stain with Weigert's iron hematoxylin for 15 minutes
• 3) Wash in running water for 15 minutes
• 4) Rinse with distilled water
• 5) Place in Van Gieson's stain for 5 minutes
• 6) Rinse in distilled water
• 7) Rinse rapidly in 70% Ethyl Alcohol
• 8) Dehydrate rapidly in Absolute Alcohol, clear and mount in Neutral Balsam. Acid medium, if used for mounting will make stains fade rapidly.

Results

• Nuclei - stains brownish black to black
• Collagen (fibrous connective tissue) - stains pink or deep red
• Muscle, Cytoplasm, RBC and Fibrin
  Fibrin
  Fibrin is a fibrous, non-globular protein involved in the clotting of blood. It is a fibrillar protein that is polymerised to form a "mesh" that forms a hemostatic plug or clot over a wound site....
  
   - stains yellow

Note:

• 1) If the duration of staining is strictly followed, there is no need to differentiate in 1% Acid Alcohol. The tissue sections are instead differentiated by Picric Acid in the Van Gieson's Stain.
• 2) Differentiation of Collagen and smooth muscles may be accomplished in steps 6 and 7. Van Gieson's Stain consists of Acid Fuchsin, which is readily soluble in alcohol, and Picric acid, which is readily soluble in water. Washing appropriately with solvent will accentuate one and remove the other.
• 3) Nuclei stained with Alum Hematoxylin is readily decolorized by Picric Acid; Iron Hematoxylin may also be used.
• 4) Young fibrils do not take the deep red stain imparted to mature collagen. RBC tends to fade after some time.

Other dyes

Other dyes used in connection with van Gieson staining include:

• Alcian blue
  Alcian blue
  Alcian blue , also called Alcian blue 8GX, Ingrain blue 1, and C.I. 74240, is a phthalocyanine . The dye stains acid mucopolysaccharides and glycosaminoglycans, for which it is one of the most widely used cationic dyes; the stained parts are blue to bluish-green...
  
  
• Amido black 10B