Laboratory Protocols

• Cryotomy
• Histological Staining Techniques
• 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

Alcian blue/PAS for acid and neutral mucopolysaccharides
Mowry, 1956.
Use a positive control section - small intestine.

• Dewax sections, rinse in alcohol, rinse in distilled water.
• Stain in 1% alcian blue in 3% acetic acid (pH2.5) - 5 minutes.
• Wash in distilled water.
• Treat with 1% periodic acid - 5 minutes.
• Wash in distilled water followed by a wash in tap water for 5 minutes then another rinse in distilled water.
• Treat with Schiff's reagent - 15 minutes.
• Wash in distilled water followed by a wash in tap water for 5 minutes then another rinse in distilled water.
• Mayer's haematoxylin - 1 minute.
• Blue in tap water.
• Dehydrate, clear and mount.

Results: 

Acid mucins - blue.
Neutral mucins - magenta.
Mixtures - purple.
Nuclei - grey/blue.