[Histonet] In Situ Hybridization Blue Haze Problems

[Brandon Dornier]

I have an In Situ Hybridization question for you guys and gals. I'm
currently performing In Situ HPV and EBV tests using an automated machine
called the XT by Ventana. The tissue is parrifin embedded, cut at 4 microns
and place on charged slides. I am beginning to notice some background
staining that varies in intensity. When it occurs on our QC slide, it can
be strong enough to fail a run. The background resembles a "blue haze" that
can cover the tissue making interpretation difficult. At other times the
blue haze surrounds the tissue rather than covering it which makes me
believe that there could be some sort of hydrophobic or other chemical rxn
taking place to shield the tissue. I'm not certain if these two problems are
related, but both occur completely random and always vary in intensity. If I
can find the culprit of this problem it would lead to cleaner slides and
more conclusive resulting. If anybody can help it would be much appreciated.
Thanks