Garvey-Movat Pentachrome
for Elastic, Mucin & 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
Alcian blue
Alcian blue 1 g
Acetic acid, glacial 3 mL
Distilled water 100 mL
Plasma stain
Biebrich scarlet, 1% aqu. 8 mL
Acid fuchsin, 1% aqu. 2 mL
Acetic acid, 1% aqu. 100 mL
Fibre stain
Saffron du Gatinais 6 g
Ethanol, absolute 100 mL
 
Add the saffron to the ethanol and seal the container.
Incubate at 56°C for 2 weeks. This solution must be anhydrous.

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. Rinse with 3% acetic acid.
  3. Place in alcian blue at 60°C for 10 minutes
  4. Rinse with distilled water.
  5. Place in working Verhoeff's solution for 6 minutes.
  6. Wash with warm tap water for 6 minutes.
  7. Place in the plasma stain for 3 minutes
  8. Rinse with distilled water.
  9. Place in the polyacid for 15 minutes.
  10. Rinse with 1% acetic acid.
  11. Dehydrate thoroughly with absolute ethanol 3 changes.
  12. Place in the fibre stain for 5-6 minutes.
  13. 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. Sections thicker than 4µ may need the elastic stain differentiated by treating with 1% aqueous ferric chloride for 20-30 seconds, then washing well with tap water.
  4. 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., (1986),
Improved Movat pentachrome stain
Stain Technology, V. 61, No 1, pp 60-62.

 


 

 

Translate in
Google Translate
Instructions