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.
Place in verhoeff's solution for 9 minutes.
Wash with warm tap water for 5 minutes.
Place in lissamine fast yellow for 2 minutes.
Rinse with 0.5% acetic acid.
Place in the plasma stain for 5 minutes.
Rinse with distilled water.
Place in polyacid for 10 minutes.
Rinse with distilled water.
Place in a fibre stain variant for 2 minutes.
Rinse with 0.5% acetic acid.
Dehydrate, clear and mount with a resinous medium.
Expected results
Nuclei – black
Elastic fibres – black
Fibrin - young – yellow
Fibrin - mature – red
Fibrin - old – blue or green
Erythrocytes – yellow
Muscle – red
Collagen – blue or green (some may be red)
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.
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.
