Protocol
Bancroft's Crystal Violet
for Amyloid
Materials
- Crystal violet, 1% aqueous
- Acetic acid, 1% aqueous
- Methyl green, 2% aqueous, washed with chloroform to remove crystal violet
Tissue Sample
5µ paraffin sections of neutral buffered formalin fixed tissue are suitable. Other fixatives are likely to be satisfactory. Cryostat sections usually show brighter metachromasia. Unmounted frozen sections may also be floated in each solution and mounted on a slide just before coverslipping.
Protocol
- Bring sections to water via xylene and ethanol, except for cryostat and frozen sections.
- Place into crystal violet solution for 5 minutes.
- Rinse with distilled water.
- Differentiate briefly with dilute acetic acid for 15 – 20 seconds.
- Counterstain with methyl green for 5 – 15 minutes.
- Wash with distilled water.
- Either drain all water from the slide until just damp then blot and mount with Apathy’s or Highman’s medium, or drain all water from the slide until just damp then blot and flood with xylene. Repeat until the section is cleared, then mount with a resinous medium.
Expected Results
- Amyloid – purple-red
- Background – green
- Nuclei – green
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- Bancroft, J. D., (1963).
Stain technology, v. 38, p. 336.London, England.
