Fig. 8. Three-dimensional reconstruction of bile canaliculi in long-term-cultured rat small hepatocytes. Cells were cultured for 7 weeks, fixed with 4% paraformaldehyde in the presence of Triton X-100 and subsequently processed for immunofluorescence localization of Mrp2. Confocal laser-scanning microscopy was used to record stacks of the cell colony that were processed for surface rendering. (Left) Immunofluorescence picture for Mrp2 of a representative stack in the middle of the z-axis. (Right) Three-dimensional view of the surface of the canalicular network after image analysis of stacked Mrp2 fluorescence. The canalicular space is shown in dark gray and the cell colony surface is shown in light gray. Scale bar, 20 µm.