Hello everyone,
I’ve been encountering high background when immunostaining paraffin-embedded, fixed tissues that include both soft and bony components.
These specimens are periapical tissues obtained from human teeth during periapical surgeries and typically contain apical periodontitis lesions (granulomas/cysts), blood vessels, nerves, periodontal ligament, tooth, and bone. When bone is present on the slide, background staining is consistently high across multiple antibodies (typically neuronal and inflammatory cell markers such as NF-H, CGRP, PGP9.5, β-tubulin III, CD45, among others). In matched sections without bone, stained using the same antibodies and protocols, background is minimal to none.
Do you have any suggestions or tips for improving the signal-to-background ratio in these mixed soft/hard-tissue sections?
Thank you very much!