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

Safranine O/fast green for cartilage

• Dewax sections, rinse in alcohol, rinse in water.
• Weigert's haematoxylin - 3 minutes.
• Wash in tap water.
• Differentiate in acid alcohol (1% hydrochloric acid in 70% alcohol) - 15 seconds.
• Wash in tap water - 3 minutes.
• Stain in 0.02% aqueous fast green - 3 minutes.
• Wash briefy in 1% acetic acid. Drain slide.
• Stain in 0.1% aqueous safranine O - 3 minutes (maximum).
• Rinse in 95% alcohol.
• Dehydrate, clear and mount.

Results:

 

Nuclei - black.
Cytoplasm - green.
Mucins, cartilage and mast cell granules - orange/red.

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