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

DMAB-rhodanine for copper
Use gelatine coated slides . Use a positive control section.

• Dewax sections, rinse in alcohol, rinse in water.
• Stain in DMAB-rhodanine stain at 60°C 3 hrs (Check microscopically after 1 hour).
• Rinse well in distilled water
• Counterstain with Mayers' haematoxylin 30 secs
• Blue in tap water - 5 minutes.
• Rinse and dehydrate in 100% alcohol.
• Clear and mount immediately (Rhodanine is gradually leached from tissue sections by the xylene in DPX so use a non-xylene mountant if possible).

Results:

 
Copper deposits - brick red
Nuclei - pale blue
Bile - green

DMAB-rhodanine, Stock solution
Make a saturated solution of the DMAB-rhodanine in 100% alcohol.
Store the stock solution at 4°C.

DMAB-rhodanine, Working solution (shake well)
DMAB-rhodanine stock solution 3 ml
Distilled water 47 ml
Prepare fresh before use.

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