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

Llewellyn, 1970.
Do not use gelatine coated slides. Use a positive control section.

• Dewax sections, rinse in alcohol, rinse in water.
• Stain nuclei with Mayer's haematoxylin - 1 minute.
• Rinse in water then 70% alcohol.
• Stain in sirius red solution - 1 hour.
• Wash in running tap water - 10 minutes.
• Dehydrate, clear and mount.

Results:

Amyloid - rose red.
Nuclei - blue.
Background - clear to pale pink.

Sirius red solution
Dissolve 0.5g sirius red in 45mls distilled water. Add 50mls 99% alcohol then 1ml 1% sodium hydroxide. Slowly add drops of 20% sodium chloride until a slight precipitate persists. (Check against a strong light. Up to 4mls may be necessary.) Leave overnight then filter. Solution is stable for 1 month only.

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