Fig 1.
Whole slide image (WSI) quantitation is shown for (A) fibrosis using trichrome, (B) basement membrane mass using Periodic acid–Schiff (PAS), (C) fibrosis using collagen III immunohistochemistry, (D) epithelial mass using the red portion of the trichrome, (E) epithelial mass using cytokeratin immunohistochemistry, and (F) using CD34 immunohistochemistry. In (A)-(F), the WSI without quantitation is seen in the upper panel; and a WSI quantitation markup image is shown in the lower panels in which yellow/orange/red areas are counted using a positive pixel count algorithm for (A)-(E) and the microvessel density is quantitated by a microvessel density algorithm in green in (F). The inset in (B) depicts the quantitation of a tubular basement membrane (green arrow) at higher power.
Table 1.
Summary of measurements obtained is shown for various measures of interstitial fibrosis, tubular atrophy, epithelial cell mass [EPCM, Epithel below], microvessel density (MVD), and microvessel area (MVA).
Stains applied include trichrome (Tri), periodic acid–Schiff (PAS), collagen III (Col) immunohistochemistry (IHC), cytokeratin (CK) IHC for EPCM, and CD34 IHC for the MVD and MVA. Measurements are either performed using morphometric or visual (Vis) methods. For fibrosis morphometry, a method employing trichrome minus the PAS measurement is used (T-P). For EPCM, the “Red” of the trichrome is also used (RedTri). The outer (Out) and inner (Inn) stripe (Stri) widths are also measured. (Std Dev = Standard Deviation).
Fig 2.
A color map of correlations between measured variables is shown.
Variables that correlate closely are clustered.
Fig 3.
Correlation of selected fibrosis measures is shown.
Image analysis of trichrome (Tri), trichrome analysis minus PAS analysis (T-P), and visual assessment (Vis) were assessed for all of the tissue, the cortex (Ctx), and the medulla (Med). Regression lines and r values of corresponding measurements show how measurements correlate. The curved dotted lines display the confidence limits for the expected mean value. Additional correlations are shown in S2 Fig.
Table 2.
Measures are shown with regard to the strength of their correlation between cortex and medulla.
Additional correlations are shown in S7 Table.
Fig 4.
Selected correlation of fibrosis measures using collagen III immunohistochemistry (Col) are shown.
Measurements were performed for all of the tissue, the cortex (Ctx), and the medulla (Med). Regression lines and r values of corresponding measurements show how measurements correlate. The curved dotted lines display the confidence limits for the expected mean value. Additional correlations are shown in S5 Fig.
Fig 5.
Selected correlation of epithelial cell mass measures are shown.
Measurements were performed for all of the tissue, the cortex (Ctx), and the medulla (Med). Regression lines and r values of corresponding measurements show how measurements correlate. The curved dotted lines display the confidence limits for the expected mean value. Additional correlations are shown in S6 Fig.
Fig 6.
Correlations of microanatomic features are shown.
(A) Correlation of outer and inner stripe width in millimeters (Out-Stri-mm and Inn-Stri-mm) with fibrosis measures, including all of the tissue, the cortex (Ctx), and the medulla (Med) using image analysis of trichrome (Tri). (B) Correlation of outer and inner stripe width in millimeters (Out-Stri-mm and Inn-Stri-mm) with measures of % epithelial cell mass [EPCM, Epithel below] by visual assessment. Regression lines and r values of corresponding measurements show how measurements correlate. The curved dotted lines display the confidence limits for the expected mean value. Additional correlations are shown in S7 Fig.
Fig 7.
Correlation of measures of microvessel density (MVD in in vessels/um2) and mean vessel area (MVA in um2) are shown between the cortex (Ctx), and the medulla (Med).
Regression lines and r values of corresponding measurements show how measurements correlate. The curved dotted lines display the confidence limits for the expected mean value. Additional correlations are shown in S8 Fig.