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

Gomori's Trichrome - one step

• Dewax sections, rinse in alcohol, rinse in distilled water.
• Stain nuclei with Celestin Blue - 5 mins
• Rinse in distilled water
• Stain in Mayers haematoxylin - 5 mins
• Wash well in running tap water - 5 mins
• Stain with Gomori’s stain - 15 mins
• Rinse with distilled water
• Dehydrate, clear and mount

Note: Step 6. Staining is progressive & should be checked after 15 mins and controlled microscopically.

Results:
Muscle fibres - red
Collagen - green
Nuclei - blue/black

Gomori’s trichrome stain
Distilled water - 200 ml
Chromotrope 2R (CI16570) - 1.2g
Light green SF(CI42095) - 0.6g
dodeca-tungstophosphoric acid  - 1.6 g
Glacial acetic acid - 2.0 mls

Mix all ingredients together.
Allow to stand overnight, filter.
The stain keeps well.

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