Garvey's Stain
for Elastic, Fibrin & Collagen

Solutions

Stock Verhoeff A
Hematoxylin 3 g
Ethanol, absolute 100 mL
Stock Verhoeff B
Ferric chloride 2.3 g
Distilled water 100 mL
Stock Verhoeff C
Potassium iodide 4 g
Iodine 2 g
Distilled water 100 mL
Working Verhoeff
Stock solution A 3 volumes
Stock solution B 2 volumes
Stock solution C 1 volume
Polyacid
Phosphotungstic acid 2 g
Distilled water 100 mL
Acetic water
Acetic acid, glacial 0.5 mL
Distilled water 100 mL
Picro-mercuric chloride
Picric acid, sat. aqu. 100 mL
Mercuric chloride 5 g
Distilled water 100 mL
Erythrocyte stain
Lissamine fast yellow 1 g
Acetic acid, glacial 0.5 mL
Distilled water 100 mL
Plasma stain
Biebrich scarlet 0.75 g
Acid fuchsin 0.75 g
Ponceau 2R 0.75 g
Acetic acid, glacial 0.5 mL
Distilled water 100 mL
Fibre stain     var I     var II
Aniline blue 0.5 g  
Light green SF yellowish   2 g
Acetic acid, glacial 1 mL 2 mL
Distilled water 100 mL 100 mL

Tissue sample
Formol sublimate fixation is preferred, although 10% formalin variants are acceptable.
Paraffin sections at 3µ are preferred.

Method

  1. Bring sections to water via xylene and ethanol.
  2. Place in verhoeff's solution for 9 minutes.
  3. Wash with warm tap water for 5 minutes.
  4. Place in lissamine fast yellow for 2 minutes.
  5. Rinse with 0.5% acetic acid.
  6. Place in the plasma stain for 5 minutes.
  7. Rinse with distilled water.
  8. Place in polyacid for 10 minutes.
  9. Rinse with distilled water.
  10. Place in a fibre stain variant for 2 minutes.
  11. Rinse with 0.5% acetic acid.
  12. Dehydrate, clear and mount with a resinous medium.

Expected results

Notes

  1. Sections should be treated with picro-mercuric chloride if formol sublimate or similar fixation was not used initially.
  2. The ferric chloride should be the hexahydrate crystals.
  3. Although the iodine content of the Verhoeff elastic stain component may remove any mercury pigment, in methods such as this some technologists prefer to apply the iodine-thiosulphate sequence before staining.

 

Reference
Garvey W., et. al., (1987),
A combined elastic, fibrin and collagen stain
Stain Technology, V. 62, No 6, pp 365-367.

 


 

 

Translate in
Google Translate
Instructions