Figure 1: The registered alternately stained histological serial section (left: five selected triple-sequences H&E–p16 INK4a –CD3 from top). Within the 3-D reconstructed tissue volume any arbitrary virtual slicing plane can be computed (middle). This image shows a plane with 1 degree tilt by this passing the same 15 consecutive sections (scale bar: 1mm, the irregular outline is due to the applied registration procedures). The upper half mainly consists of tumour invading into the stroma. While the tumour is more or less visible in all three stainings, it is best delimitable at the p16 INK4a slices as saturated brown structures. Due to the high accuracy of the obtained reconstruction, tumour structures as well as vessels and many other details smoothly continue in the adjacent slices. The spatial localisation of the inflammatory response between the tumour invasion front and the stroma as can be seen in the middle of the virtual plane is of particular tumour-biological interest. In the 3-D surface rendering (right) both the automatically segmented tumour and those tissue regions exhibiting T-lymphocytes (purple) are simultaneously shown. Distant small segments are due to staining artefacts. The overall height of the 3-D reconstructed tissue volume is 0.84mm.