The samples were represented by 85 brain samples from cattle with clinical nervous symptoms suspected of BSE.
For rapid tests and confirmatory Western blotting test the samples were represented by homogenous brainstem
tissues. The samples from brainstem, cerebellum, medulla oblongata and cerebral cortex were fixed in neutral
formalin 37% for 48 hours at room temperature. Fragments of these tissues dehydrated by successive passages in
alcohol baths of increasing concentrations were included in Paraplast Plus paraffin, were sectioned at 5 micrometers
and were staining with Haematoxilin and Eosin (H&E), Mann, Masson’s trichrome methods and
immunohistochemistry (IHC) Pullman kit for BSE.