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Van Gieson Stain For Collagen with Label

Van Gieson stain for collagen with label post we have briefly explained about Van Gieson stain method’s principle, requirements, van gieson stain procedure, result with Label.

Van Gieson Stain For Collagen with Label

Van Gieson trichrome is used for the differentiation between collagen and smooth muscle in tumors and to demonstrate the increase in the amount of collagen in these pathologies. Van Gieson Stain Procedure is composed of 3 different dyes: Weigert’s hematoxylin for nuclei, picric acid for cytoplasm and fuschine acid for collagen.

When combined solutions of picric acid and acidic fuchsin are used, small picric acid molecules penetrate rapidly into all tissues, but are firmly retained only in red blood cells and muscle. Larger fuchsin molecules displace the picric acid molecules of collagen fibers, which have larger pores, and allow large molecules to enter in van Gieson Stain Procedure.

Principle

Van gieson stain procedure is used to differentiate between collagen and smooth muscle in tumours and to demonstrate the increase of collagen in diseases.

When using combined solutions of picric acid and acid fuchsin, the small molecules of picric acid penetrate all of the tissues rapidly, but are only firmly retained in the close textured, red blood cells and muscle. The larger molecules of ponceau S displaces picric acid molecules from collagen fibres, which have larger pores, and allow the larger molecules to enter.

Requirements

Chemicals

1. Ammonium Ferric Sulphate

2. Celestin Blue

3. Picric Acid

4. Ponceau S 

 5. Glacial Acetic Acid 

Reagents

1. Celestin Blue: Add 0.5 gm of Celestin Blue (CI 51050) to the 100 ml 5% ammonium ferric sulphate (iron alum) and boil for 3 minutes. Filter when cool. Store refrigerated.

2. Curtis Stain: Mix 10.0 ml of 1% ponceau S and 10.0 ml of Glacial acetic acid with 90.0 ml of saturated aqueous picric acid.     

3. 1% Ponceau S: 1% Ponceau S is prepared by dissolving 1.0 gm of  Ponceau S (CI 27195) with 100 ml of double distilled water.      

Van Gieson Stain Procedure

1. Deparaffinize and hydrate paraffin sections in double distilled water. Bring sections to distilled water.

2. Rinse with distilled water several times. Stain nuclei with Celestin Blue 5 mins and Rinse in distilled water.

3. Stain the slides with haematoxylin for approximately for 5 mins and Wash well in running tap water 5 mins in van Gieson Stain Procedure.

4. Flood with Curtis stain 5 mins and Blot. Dehydrate the slides rapidly in alcohols, clear with xylene and mount after van Gieson Stain Procedure.

Results

Van Gieson Staining

Van Gieson Stain Procedure

Nuclei: Blue

Collagen: Bright red       

Cytoplasm, muscle, fibrin and red blood cells:  Yellow

Further Readings

Reference

  1. https://www.clinisciences.com/en/buy/cat-van-gieson-s-trichrome-stain-5387.html
  2. https://bitesizebio.com/19952/verhoeff-van-gieson-stain-a-special-histology-stain-for-elastic-fibers/