Figure 4. Distribution of hepatocyte volumes and DAPI integral intensity per cell for all hepatocytes (A, B) and separated by number of nuclei (B, C and E, F).
Whereas experimental data are shown by dots, the log-normal components fitted to data are shown by solid lines. (A) Cell volume distribution of all hepatocytes. (B, C) Cell volume distribution obtained for mono and bi-nucleated hepatocytes, respectively. (D) Distribution of DAPI integral intensity (proportional to the content of DNA) of all hepatocytes. (E, F) Distributions of DAPI integral intensity obtained for mono and bi-nucleated hepatocytes, respectively. The analysis was performed on 2559 hepatocytes (excluding boundary cells) from three adult mice.
Figure 4—figure supplement 1. Morphometric features of the sinusoidal and bile canalicular (BC) networks.
(A) Radius distribution of the sinusoidal capillary network. (B) Distributions of the angles between branches of BC and sinusoidal networks. (C) Cardinality of branching nodes of BC and sinusoidal networks. The data shown here correspond to a representative sample of adult mouse liver.
Figure 4—figure supplement 2. (A, B) DAPI integral intensity normalization.
Distribution of DAPI integral intensity per nucleus calculated for each sample (A) before and (B) after normalization. We found scaling (stretching) factors 1.19 and 0.93 for the second and third samples, respectively. (C, D) DNA content in bi-nuclear hepatocytes. DAPI integral intensity per nucleus was calculated for each nucleus of the cells. (C) The distribution of the ratio between DAPI integral intensity of the two nuclei in each cell. It follows a normal distribution with mean value 1.0 ± 0.21 (mean ± SD). (D) The dependency between DAPI integral intensity of the two nuclei in bi-nuclear cells. They show a linear dependency (R2 = 0.945433) with a slope of 0.995, showing that both nuclei have the same DNA content in bi-nuclear hepatocytes. (E, F) Scatter plot of the volume versus DAPI integral intensity of (E) mono-nuclear and (F) bi-nuclear hepatocytes. The results of the hierarchical clustering of (E) mono-nuclear and (F) bi-nuclear hepatocytes are shown. Four (2n, 4n, 8n, 16n) and three (2×2n, 2×4n, 2×8n) populations were found for mono-nuclear and bi-nuclear hepatocytes, respectively. The classification was performed using volume and DAPI integral intensity per cell. We used an agglomerative hierarchical cluster algorithm and tested several distances for the dissimilarity calculation and different methods for the clustering. We found that the standardized Euclidean distance with the Ward method yielded the best results.