Mice were sacrificed shortly after the last MRI acquisition. All livers were macroscopically analyzed for focal hepatic nodules. Before liver sampling, the orientation of all liver lobes, relative to the longitudinal, transversal and sagittal axis of the animal, was marked, and the size and location of the macroscopic tumor nodes were measured with a caliper and correlated with the MRI findings. Part of the respective liver was then embedded in paraffin and used for diagnostic procedures. The rest of the liver was frozen for further analysis.
All tissue samples were fixed in 10% buffered formalin for 24 h and paraffin embedded. For histological analysis, 4-μm-thick serial sections were stained with Hematoxylin and Eosin. Reticulin was visualized by Gomori staining. Immunostaining was performed with anti-GS (Zulehner et al., 2010 (link)) (1:500; ab49873, Abcam, Cambridge, UK) and anti-SAA (1:100; PAB795Mu01, Cloud-Clone, Houston, TX, USA) antibodies. Reactions were developed using an EnVision+ System-HRP (DAB) (DAKO, Carpinteria, CA, USA). After immunostaining, slides were counterstained with Hematoxylin. Positive and negative controls were included for each run.
All tissue samples were fixed in 10% buffered formalin for 24 h and paraffin embedded. For histological analysis, 4-μm-thick serial sections were stained with Hematoxylin and Eosin. Reticulin was visualized by Gomori staining. Immunostaining was performed with anti-GS (Zulehner et al., 2010 (link)) (1:500; ab49873, Abcam, Cambridge, UK) and anti-SAA (1:100; PAB795Mu01, Cloud-Clone, Houston, TX, USA) antibodies. Reactions were developed using an EnVision+ System-HRP (DAB) (DAKO, Carpinteria, CA, USA). After immunostaining, slides were counterstained with Hematoxylin. Positive and negative controls were included for each run.
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