Problems in Histopathological Technique

 

Prepared by

ROY ELLIS

IMVS Division of Pathology

The Queen Elizabeth Hospital

Woodville Road, Woodville, South Australia 5011

Email: roy.ellis@imvs.sa.gov.au

 

 

 

PROBLEM NUMBER 24

What is the best way to destain a slide to be stained by another technique?


One solution:


This solution is a fairly common bleaching protocol used in many silver techniques but it is one which also works well for bleaching stained sections and most staining methods are possible after its use.

  • First remove the coverslip and rehydrate the section.

  • Then treat with aqueous 0.025% potassium permanganate for 1 minute

  • Then rinse in water

  • followed by 5% aqueous sodium thiosulphate until the section appears bleached. Or 1% oxalic acid can be used instead of thiosulphate.

  • Wash in water for 5 minutes then re-stain

It is important not to overdo the oxidation step with potassium permanganate because overoxidation is like overfixation, if you go too far then you will never restain the section well. And if the section doesn’t properly bleach the first time you can always go back and do it again - which is much the better option than overoxidation.

 

 

 

Click the button to return to the Home Page

© Roy C. Ellis 2002