Figure 3

Three-dimensional reconstruction of injured rat liver mounting a HPC response. Three-dimensional (3D) reconstructions of 208 μm rat liver tissue demonstrated protein expression in control liver (A) and liver from day 9 in the AAF/PHx protocol (B). Hepatic tissues were visualized by (a) volumetric rendering presenting the immunohistochemistry and (d,f,h) segmentation based reconstructions of lumina and protein expression. These techniques are presented combined in subpanels b, c, e, g, i, and j. In control rat liver (A, a-j) and day 9 in the AAF/PHx protocol (B, a-j) two portal areas merged in one end of the image stack (d) and continued as separate entities. Presented in green, lumina of slender portal bile ducts (b) along the portal vein connecting the portal areas could be traced in both livers. Whereas Dlk1 localized peripherally to the portal area in the AAF/PHx protocol (B;c,d) and was undetectable throughout the image stack from control liver (A;c,d), HAI-1 marked the epithelial cells in the entire biliary tree (A, e and f) and the HPC response (B;e,f). In control liver (A;g,h) Nidogen1 was only expressed in the portal area and circumscribed the biliary tree, and additionally embedded the HPC response in the AAF/PHx protocol (B;g,h). Combined staining of HAI-1 and Nidogen1 emphasized nidogen1 deposition around the HAI-1, expressing cells in the bile epithelium (A,B;i), and clarified Dlk1 as a marker of a subpopulation of HAI-1 expressing cells within the HPC response (B;j).