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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
Muddy background staining when using Verhoeff’s elastic stain with a Van Gieson counterstain.
Verhoeff’s stain is notoriously difficult to differentiate so that fine and coarse elastic fibres can be seen. Picric acid in the van Gieson counterstain also differentiates Verhoeff as well as ferric chloride which is also used in the method. Residual ferric chloride is responsible for the muddy background which is even muddier if all of the Verhoeff’s stain is not removed from the cytoplasm.
So one solution is to differentiate the elastic fibres with picric acid instead of ferric chloride the elastic fibres are as easy to differentiate and the background is certainly cleaner than when ferric chloride is also used. But if you are going to demonstrate elastic fibres the Millers stain is easier to control and there is no background staining.
This is an elastic stain using Verhoeff’s solution and ferric chloride as a differentiator.
This is the same section after using only picric acid as a differentiator - the elastic fibres are definitely crisper.
This is a section stained with Miller’s elastic stain and counterstained with picric acid - in this section even the fine elastic fibres stand out well against the yellow background.
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© Roy C. Ellis 2002