Laboratory Protocols

• Cryotomy
• Histological Staining Techniques
  • Index
• Histological fixatives
• Peroxidase-anti-peroxidase Technique
• Preparation of tissue section adhesives
• Processing tissues for light microscopy (basics) (PDF 1.1mb)
• Processing tissues for light microscopy (detailed)
• Sample collection guidelines
• Setting up the light microscope
• Specimen preparation for electron microscopy

Histological Staining Techniques

Rubeanic acid for copper
Howell, 1959.
Use gelatine coated slides or celloidinise. Use a positive control section.

• Dewax sections, rinse in alcohol, rinse in distilled water.
• Treat in a coplin jar containing 2.5mls 0.1% rubeanic acid (dithio-oxamide) and 50mls of 10% sodium acetate at 37^oC for 4-8 hours.
• Wash briefly in distilled water and blot dry.
• Counterstain with nuclear fast red - 1 minute.
• Wash in distilled water.
• Dehydrate, clear and mount.

Results: 

Copper rubeanate - greenish black.
Nuclei - red.

*Make the test solution up fresh each time.

Stock solutions
Rubeanic acid (dithio-oxamide) = 0.1g rubeanic acid in 100ml methylated spirit.
Sodium acetate = 10g anhydrous sodium acetate in 100ml distilled water.

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