I am doing In Situ Hybridization using HPV16/18 or HPV14 probes for human Cervical cancer FFPE tissue.
As the negative tissue does not give any positive signal, but positive signal give strong nuclear staining.
Only the issue is that the signal is penetrating the basement membrane. (even the cervical cancer is not invasive).
I know there are few possibility including the probe specificity, stringency wash is not sufficient, or non-specific signal from the signal amplification methods, but are there any way those signal could be real?
Does anyone know any reference showing the HPV signal can be observed other than basement membrane and epithelium?
Thank you in advance.