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

Grimelius, 1968.

Use a positive control section.

• Dewax sections, rinse in alcohol, rinse in distilled water.
• Transfer to preheated silver solution at 60^oC - 3 hours.
• Drain silver from the slides and place in freshly prepared reducer at 45^oC for 1 minute*
• Rinse in distilled water.
• Counterstain with 1% light green in 1% acetic acid - 1 minute.
• Wash in tap water for 2-3 minutes to differentiate the green.
• Dehydrate, clear and mount.

*If the stain is not dark enough the slides may be put back into the silver after a wash in distilled water.

Results: 

Carcinoid tumours (shown), Argyrophil cells, A cells of pancreatic islets - brown/black.
Background - green shades.
Nuclei - light brown.

Silver solution

Acetate buffer pH5.6 - 10mls
Distilled water - 87mls
1% silver nitrate - 3mls

Reducer

Hydroquinone (quinol) - 1g
Sodium sulphite - 5g
Distilled water - 100mls
Prepare just before use.

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