[Histonet] Evans Blue and immunhistochemistry

[Foerster, Katharina]

Dear Histonetters,

I`m looking for information about the parallel use of Evans Blue (to 
identify the area at risk induced by coronary artery ligation) and 
TTC (to identify the infarct zone) with a following immuno staining 
for vessels (anti-smooth-muscle-actin-antibody and cd31).
Additionally, I wish to use Haematoxylin-Eosin staining and 
Elastica-van-Gieson in the heart-tissue.

Evans Blue and TTC are necessary for macroscopic infarct size 
assessment and not replacable in this project.
But it will be possible to take the tissue samples before TTC 
staining, so only Evans Blue (4 - 5 ml 10% dye per vena jugularis 
of the rabbit) remains problematical. 

I found some information about Evans Blue and H & E staining and 
this combination seems to be possible.

But I`m not sure if Evans Blue interferes with or covers any of the 
other stainings. 
If so, is there a possibility to wash out Evans Blue before 
immunhistochemistry (heart tissue will be fixed in 4% 
formaldehyde before embedded in paraffin)? Or is the microscopic 
tissue discolouration negligible because of albumin-binding?

Thank you for your help.
Katrin.



Katharina Förster
University Hospital Freiburg
Dept. Heart and vessel surgery
79106 Freiburg
Germany
e-mail: foerster <@t> ch11.ukl.uni-freiburg.de