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

High iron diamine/alcian blue for sulphated and carboxylated mucins (HID/AB)

Spicer, 1965.

Use a positive control section.

• Take sections to distilled water.
• Treat in HID solution - 16 hours at room temperature.
• Wash well in running tap water.
• Stain in 1% alcian blue in 3% acetic acid (pH2.5) - 5 minutes.
• Rinse in tap water.
• Dehydrate, clear and mount.

Results:

Sulphated mucins - brown/black.
Carboxylated mucins - blue.

HID solution

N,N-dimethyl-meta-phenylenediamine-dihydrochloride - 120mg
N,N-dimethyl-para-phenylenediamine-dihydrochloride - 20mg
Distilled water - 50mls
Freshly prepared 60% ferric chloride solution - 1.4mls

Dissolve the two diamine salts in the distilled water and then add the ferric chloride solution. Make the solution up directly into a Coplin jar for staining.

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