Figure 8

Individual cell annotation allows quantification of fine-grain cellular relationships that derives new insights into fundamental processes in translational models of disease. (a) Early pericentral hepatic fibrosis (picro-sirius red-stained, scale bar 1 mm), was induced in a cohort of wild type mice (n = 6), and sections stained for αSMA to identify MFBs (b, lilac, scale bar 100 μm). (c) 10 pericentral fields from each were used to annotate the nuclear position of each MFB, and the circumference of the central vein, to generate spatial point patterns from which the distances (d, red) of individual cell from vessels (lilac) and relative polar angle of individual cells with respect to vessel lumen centroids (ϕ, blue) could be calculated. (d) Scar phenotyping by density distribution of calculated MFB-central vein distances (d) for each animal demonstrates an MFB gradient within scars, highest at the central veins. (e) The nuclear position of each MFB was converted to polar coordinates with reference to the calculated centroid of the annotated vessel. The polar angle of the peak of the kernel density estimate of all polar MFB angles was set to 90° by ‘rotating’ all MFBs about the central vein centroid to allow alignment of all images. (f) Aggregates of aligned MFBs plotted for each animal as a polar histogram or kernel density estimate demonstrated a dominant pericentral spur with a smaller secondary antipodal spur. (g) The data can be fitted by a sine wave with 180° equivalent periodicity. (h) Diagrammatic representation of normal liver lobules with the classic hexagonal arrangement, and diffuse pericentral fibrosis or with dominant and single antipodal spurs; visual comparison of identical murine liver stained with picro-sirius red shows that liver scarring is organised with a dominant scarring axis, accompanied by a single secondary directly-opposed axis rather than developing uniformly along all available central-central axes (scale bar 500 μm).