Toluidine Blue
for Nissl bodies


Staining Solution Or
Toluidine blue 1 g
Sodium tetraborate 1 g
Distilled water 100 mL
Staining Solution
Thionin 0.1 g
Distilled water 100 mL
Gothard's Differentiator
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.


  1. Bring sections to water via xylene and ethanol.
  2. Place into the staining solution at 56°C for at least 30 minutes
    or up to overnight at room temperature.
  3. Rinse well with running tap water.
  4. Rinse with absolute ethanol.
  5. Differentiate with Gothard's differentiator, controlling microscopically.
  6. Rinse well with absolute ethanol.
  7. Clear with xylene and mount using a resinous medium.

Expected results


  1. Disbrey omits the sodium tetraborate from the toluidine blue solution and stains at room temperature, but recommends overnight staining when possible.
  2. Methylene blue may be substituted for thionin.
  3. Culling recommends diluting the Gothard's differentiator with an equal volume of absolute ethanol before use.


Wikipedia articles   Cajeput oil   Creosote

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.




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