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

Wright 1928.
Prepare cytospin slide or smear of sample.

• Stain air-dried smears in a closed coplin jar of Wright's stain - 5 minutes.
• Place the slide on a rack, rinse and treat with pH7.2 buffer primed with 1ml Wright's stain per 400ml - 3 minutes.
• Rinse in pH7.2 buffer - 20 seconds.
• Blot dry, clear and mount.

Results:

Red blood cells - red to pink.

• Neutrophils - dark purple nuclei, pale pink cytoplasm, reddish-lilac small granules
• Eosinophils - blue nuclei, pale pink cytoplasm, red to orange-red large granules
• Basophils - purple to dark blue nucleus, dark purple, almost black large granules
• Lymphocytes - dark purple to deep bluish purple nuclei, sky blue cytoplasm
• Platelets - violet to purple granules

Wright's stain
6.0g Wright's stain powder
0.6g Giemsa stain powder
1,000ml methanol
Stir overnight and filter before use.

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