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

Wade 1957.
Do not celloidinise. Use a positive control section.

• Dewax the sections in a mixture of equal parts of liquid paraffin and oil of turpentine for at least an hour. Sections can be left in this mixture overnight to make sure all the wax is removed.
• Blot until almost dry.
• Wash in running tap water - 5 minutes.
• Stain in carbol-fuchsin on a rack at room temperature - 30 minutes.
• Wash in running tap water, blot dry.
• Decolourise with 10% sulphuric acid - 2 minutes.
• Wash in running tap water.
• Counterstain with Mayer's haematoxylin - 1 minute.
• Wash and blue in running tap water - 10 minutes.
• Blot dry and complete drying on a 60^oC hotplate.
• Clear in Histo-Clear^® or xylene and mount.

Results:

M. leprae (and other mycobacteria) - red
Nuclei - grey/blue

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