Histological Analysis of Mouse Liver

After fixation in 10% neutral-buffered formalin over 24 h, the same liver lobe of all mice was selected, embedded in paraffin, cut into sequential sections of 3–5 μm in thickness and subjected to Hematoxylin-Eosin (HE) staining and immunohistochemistry (IHC). The frozen sections were cut into 5- to 8-μm sequential sections and subjected to Oil Red O staining (Oil Red O Stain Kit; Polysciences, Inc., Warrington, PA, USA). All section images were captured with the Nano Zoomer Digital Pathology Virtual Slide Viewer software program (Hamamatsu Photonics Corp, Hamamatsu, Japan). As described in a previous study [11 (link)], we used the NAFLD score (steatosis score and ballooning score) to assess the condition of the mouse liver (Table 1).