Histopathological Analysis of Spleen and Liver

Spleens and livers were weighed and then fixed in 10% buffered formalin and processed into paraffin by the Histopathology Department at the Royal (Dick) School of Veterinary Studies using standard procedures. Slides were stained with hematoxylin and eosin (H&E). Immunohistochemistry (IHC) was performed with mouse anti-rat CD68 (Clone ED1, 1:500, Bio-Rad). For Oil red O staining, formalin-fixed livers were placed in 18% sucrose at 4°C overnight and cryosections prepared as described in (45 (link)). Staining was performed as described in Ref. 29 (link). Sections were imaged using a Nanozoomer digital scanner and viewed using NDP.view 2 (Hamamatsu, Japan). Oil red O staining was imaged with standard light microscopy using ZEN software (Zeiss).