Toluidine Blue
for Nissl bodies
Materials
- Staining Solution
- Option 1
Material Amount Toluidine blue 1 g Sodium tetraborate 1 g Distilled water 100 mL - Option 2
Material Amount Thionin 0.1 g Distilled water 100 mL
- Option 1
- Gothard’s Differentiator
Material Amount Creosote 50 mL Cajeput oil 40 mL Xylene 50 mL Ethanol, absolute 160 mL
Tissue Sample
10µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory.
Protocol
- Bring sections to water via xylene and ethanol.
- Place into the staining solution at 56°C for at least 30 minutes
or up to overnight at room temperature. - Rinse well with running tap water.
- Rinse with absolute ethanol.
- Differentiate with Gothard’s differentiator, controlling microscopically.
- Rinse well with absolute ethanol.
- Clear with xylene and mount using a resinous medium.
Expected Results
- Nissl bodies – blue
- Nuclei – blue
- Background – pale to unstained
Notes
- Disbrey omits the sodium tetraborate from the toluidine blue solution and stains at room temperature, but recommends overnight staining when possible.
- Methylene blue may be substituted for thionin.
- Culling recommends diluting the Gothard’s differentiator with an equal volume of absolute ethanol before use.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Disbrey, B. D., (1970)
Histological laboratory methods, p. 232.
E. & S. Livingstone, Edinburgh and London, UK. - Culling, C F A, Allison, R T, Barr, W T, (1985).
Cellular pathology technique., Ed. 4.
Butterworths, London, England.
