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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
What is the best way to destain a slide to be stained by another technique?
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.
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First remove the coverslip and rehydrate the section.
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Then treat with aqueous 0.025% potassium permanganate for 1 minute
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Then rinse in water
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followed by 5% aqueous sodium thiosulphate until the section appears bleached. Or 1% oxalic acid can be used instead of thiosulphate.
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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.
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© Roy C. Ellis 2002