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

Dunn/Thompson for haemoglobin
Dunn & Thompson, 1945.
Use a positive control section.

• Dewax sections, rinse in alcohol, rinse in water.
• Stain in a mixture of equal parts 1% alcoholic haematoxylin (ripened, not fresh) and 10% potassium alum - 15 minutes. This solution may crystallise but is still usable.
• Rinse briefly in tap water.
• Mordant in 5% iron alum - 1 minute.
• Rinse briefly in tap water.
• Restain in the solution used at the second step - 10 minutes.
• Rinse briefly in tap water.
• Stain in van Gieson's stain - 15 minutes.
• Differentiate in 95% alcohol - 3 minutes.
• Dehydrate, clear and mount.

Results: 

Red blood cells - greenish black.
Haemoglobin castes or phagocytosed haemoglobin - lighter shades of green.
Collagen - red.
Muscle and cytoplasm - yellow.

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