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

Schmorl for lipofuscins
Golodetz and Unna, 1909.
Use a positive control section.

• Dewax sections, rinse in alcohol, rinse in distilled water.
• Incubate at room temperature for 1 minute (agitate the slides).
• Wash in distilled water and examine the control section for staining.
• Repeat, if necessary, for up to 5 minutes in 30 second steps.
• Wash well in tap water.
• Treat with 1% acetic acid - 5 minutes.
• Wash in distilled water.
• Counterstain with nuclear fast red - 5 minutes.
• Wash in distilled water.
• Dehydrate, clear and mount.

Results:

Lipofuscins, (also melanin, liver bile, haematoidin, argentaffin cells, chromaffin cells, sulphydryl groups) - varying shades of blue/black.
Nuclei - red.

Incubating solution
1% ferric chloride (dilute 'Analar' 60%*) - 37.5mls.
1% potassium ferricyanide (fresh)- 5mls.
Distilled water- 7.5mls.
*0.625mls 'Analar' ferric chloride in 37.5mls distilled water.

Make the incubating solution up fresh and use within 30 minutes.

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