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. 2020 Feb 3;26(2):80–90. doi: 10.1089/ten.tec.2019.0228

FIG. 3.

FIG. 3.

Distribution of cell types within spheroids. Image size: 400 × 400 μm. (A–F) Troponin T stain for cardiomyocytes. Green = Troponin T, blue = DAPI. (G–L) Vimentin stain for fibroblasts. Green = Vimentin T, blue = DAPI. (A, D, G, L) Group 1 spheroid (CM:FB:HUVEC 70:15:15). (B, E, H, K) Group 2 spheroid (CM:FB:EVC 70:15:15). (C, F, I, L) Group 3 spheroid (CM:FB:EVC 40:15:45). (M, N) Illustration depicting method of generating inner/outer ratio using Troponin T and vimentin stains. (M) Normalized integrated fluorescence intensity along concentric circles was obtained with the ImageJ Radial Profile Plot plugin. (N) The plot was divided at half-radius corresponding to an “inner core” (blue) and “outer shell” (yellow) for the spheroid. Dividing the resulting sums of integrated intensities gives the “inner/outer ratio.” (O, P) Inner/outer ratio obtained from (O) Troponin T stains for cardiomyocytes, and (P), vimentin stains for fibroblasts. (Q, R) Percent composition of (Q) Troponin T-positive cells and (R) vimentin-positive cells derived from normalized integrated fluorescence intensity data. On bar charts, *p < 0.05, **p < 0.01.