Histological Staining Techniques
Puchtler, Sweat, Levine, 1962.
Cut sections at 6µm. Do not use gelatine coated slides. Use
a positive control section.
Dewax sections, rinse in alcohol, rinse in water.
Stain in Harris haematoxylin - 10 minutes.
Wash in tap water - 1 minute.
Differentiate in acid alcohol - 10 seconds.
Blue in tap water - 5 minutes.
Treat with stock solution A - 20 minutes in a sealed coplin jar.
Treat with stock solution B - 20 minutes in a sealed coplin jar.
Rinse and dehydrate in 100% alcohol.
Clear and mount.
Amyloid, eosinophils, elastin - brick red.
Amyloid also displays apple-green birefringence under polarized light.
Nuclei - blue.
Background - clear/pale pink.
Stock solution A
Saturated sodium chloride in 80% alcohol
Working solution = 50ml stock solution A + 0.5ml 1% aqueous sodium hydroxide
Stock Solution B
Saturated congo red in 80% alcohol saturated with sodium chloride
Working solution = 50ml stock solution B + 0.5ml 1% aqueous sodium hydroxide
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