[Histonet] Tissue staining artefacts - areas of tissue not labelling

[Peter March]

Hi,

A lab which does a lot of immuno work on tissue sections (both DAB and fluorescent labelling) have run into issues with uniformity of antibody staining. Some areas of the tissue just don’t label and it seems to be a very clear line between the stained and unstained tissue. The tissue was sent to them by a collaborator as a FFPE wax block and they section from this block. They are meticulous with their processing and make sure the tissue never dries out during processing or the PAP pen marker is nowhere near the tissue. There is no issue with H&E staining and no issue with the DAPI labelling, just when they use antibodies. Does this look like an issue with the processing of the tissue before embedding or a downstream issue? Any suggestions would be very welcome.

I’ve uploaded some images to show the artefacts. The staining in all should be uniform across the tissue.

https://imgur.com/gallery/HtaQP5p

Thanks,

Peter

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Dr Peter March
Senior Experimental Officer
Faculty of Biology, Medicine and Health
The Michael Smith Building
The University of Manchester
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Manchester
M13 9PT

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