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

M.P.T.A.H. (Mallory's phosphotungstic acid haematoxylin) for glial fibres

Mallory 1897, 1900.

• Dewax sections, rinse in alcohol, rinse in water.
• Treat in 0.25% potassium permanganate - 5 minutes.
• Wash in distilled.
• Bleach in 1% oxalic acid - 2 minutes.
• Rinse in distilled, wash in tap 2 minutes, rinse in distilled.
• Stain in P.T.A.H. - 24 hours in a coplin jar.
• Treat in 95% alcohol - 2 minutes.
• Dehydrate in 100% alcohol.
• Clear and mount.

Results

: 

Glial fibres, nuclei, fibrin, myelin - blue.
Neurones - pink.
Collagen, reticulin - brick red.
Background - pale pink.

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