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

Page, 1965, 1970.
Use 7-10µm sections, or 10-15µm sections for central nervous system (C.N.S.)

• Dewax sections, rinse in alcohol, rinse in water.
• Stain with solochrome cyanine - 10 minutes.
• Blue in tap water - 5 minutes.
• Differentiate in 10% iron alum for 1-2 minutes until the nuclei are pale. Check under the microscope.
• Rinse well in running water.
• Counterstain central nervous system (CNS) sections in van Gieson stain - 2 minutes. Omit this step for other tissues.
• Rinse in distilled (blot dry for van Gieson).
• Dehydrate, clear and mount.

Results: 

Peripheral nerve.
Eosinophil granules, myelin, red blood cells, muscle, nuclei (in descending order of intensity) - blue.

Results:

C.N.S.
Counterstained with van Gieson, as above but with collagen - red and cytoplasm - yellow.

Solochrome cyanine solution
Add 0.5ml conc. sulphuric acid to 0.2g solochrome cyanine RS in a flask. Stir well with a glass rod until dissolved. Add 96mls distilled water and 4mls 10% aqueous iron alum. Mix and filter. Keeps well.

Note:
Solochrome cyanine RS is also referred to as Eriochrome cyanine R (both are C.I. number 43820).

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