Cresyl Violet Stain
for Nissl Bodies

Solutions

Staining Solution
Cresyl violet 1 g
Distilled water 100 mL
Differentiator Or
Ethanol, 95% 100 mL
Cajeput oil a few drops
Differentiator
Ethanol, 95% 100 mL
Acetic acid, glacial 5 drops

Tissue sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives, particularly if ethanolic, are likely to be satisfactory.

Method

  1. Bring sections to water via xylene and ethanol.
  2. Place into the staining solution for 15-30 minutes.
  3. Rinse with water, dehydrate with absolute ethanol and clear with xylene.
  4. Leave in xylene for one hour.
  5. Rinse well with absolute ethanol.
  6. Bring one slide at a time to 95% ethanol.
  7. Differentiate in either cajeput ethanol or acetic ethanol, controlling microscopically.
  8. Rinse well with 95% ethanol to stop differentiation.
  9. Dehydrate with absolute ethanol.
  10. Clear with xylene
  11. Coverslip using a resinous medium.

Expected results

Notes

  1. The first dehydration and clearing of the stained section (steps 3-4) improves the sharpness of diffentiation and is recommended.
  2. Differentiation, as in steps 5-10, may be repeated if necessary.

 

Reference
Wikipedia articles   Cajeput oil

Drury, R A, and Wallington, E A, (1967).
Carleton's histological technique., Ed. 5., pp. 379.
Oxford University Press, London, England.

Bench Manual,
Dept of Pathology, Prince George Regional Hospital,
Prince George, BC, Canada

 


 

 

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