Bancroft's Crystal violet
for Amyloid

Solutions

· 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.

Method

  1. Bring sections to water via xylene and ethanol, except for cryostat and frozen sections.
  2. Place into crystal violet solution for 5 minutes.
  3. Rinse with distilled water.
  4. Differentiate briefly with dilute acetic acid fro 15 - 20 seconds.
  5. Counterstain with methyl green for 5 - 15 minutes.
  6. Wash with distilled water.
  7. Either
    • Drain all water from the slide until just damp then blot and mount with Apathy's or modified Apathy'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

Notes

  1. In this method the methyl green is used as a counterstain, so it must have any contaminating crystal violet removed with chloroform washes.

 

Reference
Bancroft, J. D., (1963).
Stain technology, v. 38, p. 336.
London, England.

 


 

 

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