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
Bring sections to water via xylene and ethanol.
Optionally, place into picro-mercuric chloride for 1 hour.
Wash well with warm water to remove picric acid.
Rinse with 3% acetic acid.
Place in alcian blue at 60°C for 10 minutes
Rinse with distilled water.
Place in working Verhoeff's solution for 6 minutes.
Wash with warm tap water for 6 minutes.
Place in the plasma stain for 3 minutes
Rinse with distilled water.
Place in the polyacid for 15 minutes.
Rinse with 1% acetic acid.
Dehydrate thoroughly with absolute ethanol 3 changes.
Place in the fibre stain for 5-6 minutes.
Dehydrate, clear and mount with a resinous medium
Expected results
Nuclei – black
Elastic fibres – black
Fibrin - mature – red
Muscle – red
Collagen – yellow
Ground substance – blue-green
Notes
Sections should be treated with picro-mercuric chloride if formol sublimate or similar fixation was
not used initially.
The ferric chloride should be the hexahydrate crystals.
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.
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.
