Cresyl Violet Stain
for Nissl Bodies
Materials
Staining Solution
| Material | Amount | |
|---|---|---|
| Cresyl violet | 1 | g |
| Distilled water | 100 | mL |
Differentiator – Option 1
| Material | Amount | |
|---|---|---|
| Ethanol, 95% | 100 | mL |
| Cajeput oil | a few drops | |
Differentiator – Option 2
| Material | Amount | |
|---|---|---|
| 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.
Protocol
- Bring sections to water via xylene and ethanol.
- Place into the staining solution for 15-30 minutes.
- Rinse with water, dehydrate with absolute ethanol and clear with xylene.
- Leave in xylene for one hour.
- Rinse well with absolute ethanol.
- Bring one slide at a time to 95% ethanol.
- Differentiate in either cajeput ethanol or acetic ethanol, controlling microscopically.
- Rinse well with 95% ethanol to stop differentiation.
- Dehydrate with absolute ethanol.
- Clear with xylene
- Coverslip using a resinous medium.
Expected Results
- Nissl bodies – blue-violet
- Nuclei – blue violet
Notes
- The first dehydration and clearing of the stained section (steps 3-4) improves the sharpness of diffentiation and is recommended.
- Differentiation, as in steps 5-10, may be repeated if necessary.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Drury, R A, and Wallington, E A, (1967).
Carleton’s histological technique., Ed. 5., pp. 379.
Oxford University Press, London, England. - Bench Manual
