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

Naumenko/Feigin (modification of Cajal) for astrocytes

Naumenko, Feigin, 1936.
Use 15µm sections.

• Dewax sections, rinse in alcohol, rinse in water.
• Place in F.A.B. for 3 days at room temperature in a sealed glass coplin jar.
• Rinse in distilled water.
• Place slides in gold solution overnight in the dark. Prepare the solution just before use and use plastic forceps to transfer the slides into it.
• Rinse in distilled water - 2 minutes.
• Fix in sodium thiosulphate - 5 minutes.
• Rinse in distilled water.
• Dehydrate, clear and mount.

Results:

 
Astrocytes - black.
Background - pale yellow to red/pink.

F.A.B. solution
Ammonium bromide - 0.6g
Formalin (not neutral buffered) - 14mls
Distilled water - 100mls

Gold solution
1% gold chloride - 8mls.
Distilled water - 40mls.
5% mercuric chloride - 6.4mls.
Use a glass or plastic stirrer to mix.

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