Immunohistochemical studies on meningoencephalitis in feline infectious peritonitis (FIP)

WANG, Huanan; HIRABAYASHI, Miyuki; CHAMBERS, James K.; UCHIDA, Kazuyuki; NAKAYAMA, Hiroyuki

All tissues were fixed in 10% neutral-buffered formalin, routinely processed and embedded in paraffin wax. Tissue sections of 4 µm-thick were stained with hematoxylin and eosin (HE). Selected brain sections were subjected to immunohistochemical examinations. Normal feline brain tissue was also stained as a control. Deparaffinized and rehydrated sections were treated through the antigen retrieval methods listed in Table 2 . After being immersed in 3% hydrogen peroxide in methanol at room temperature for 5 min, the sections were treated with 8% skim milk at 37°C for 40 min. The sections were then incubated with the primary antibodies listed in Table 2 at 4°C overnight, and then incubated with the Dako EnVision + System horseradish peroxidase (HRP)labelled anti-rabbit (Dako-Japan, Tokyo, Japan) or anti-mouse secondary antibodies (Dako-Japan) at 37°C for 40 min. The antigenantibody reaction was visualized with 0.05% 3.3′-diaminobenzidine and 0.03% hydrogen peroxide in tris-hydrochloric buffer. Counterstaining was performed with hematoxylin.