[Histonet] Weak Hematoxylin Staining?

[Vivian Hou]

Dear all,

We are doing some immunofluorescent stains that we want to pair with H&E
slides. We are using fresh frozen sections for both (unfixed tissue, human
diseased coronary arteries) but I am seeing very weak hematoxylin staining
to the point where I have to dial back the eosin (3 min hematoxylin, 1 dip
eosin).


I am using Gills 2 (newly purchased), I know its not necessary to filter it
but we have been just in case using Whatman no. 5 filters. No
differentiation due to the weak staining right now.


I would appreciate any thoughts on why the staining might be so weak, we do
a great deal of processing and imaging on the arteries (about 12 hrs @ 25C)
prior to histo and I wonder if this has cause the tissue to decay?


Thank you for your time,
V

-- 
-----------------------------------------------------------------------------------------
V Hou
Bioengineering | Human Photonics Laboratory
e: houv2 at uw.edu | c: 206-999-3708