Fig. 3
From: Multiscale cytometry and regulation of 3D cell cultures on a chip
Multiscale cytometry on chip. a Measured amplitude of intracellular ALB. Each dot represents one spheroid and each color represents one chip. The mean value for each chip is shown by a larger dot with the same color. Solid black line: average ALB signal at each time point. D + 1/D + 3/D + 5/D + 7: n = 5/4/6/4 chips; N = 4,925 spheroids. b, c Normalized ALB signal for each spheroid vs. b the spheroid diameter and c the shape index. The normalization was performed by dividing a specific spheroid ALB intensity value by the average ALB signal within a chip. By this way, it was possible to compare ALB production of spheroids from different chips. The histograms show the distribution of the data along the different axes. Blue curves show Gaussian fits, dotted blue line: mean, solid blue line: median. N spheroids = 4,925, N cell aggregates = 506, and N units = 389. d Representative images for a spheroid in bright field (top), stained for DAPI (middle), and for ALB (bottom). R represents the effective spheroid radius and r the radial coordinates of each detected peak in the fluorescent signal within the spheroid. Scale bar is 50 μm. e Mean distance between each cell and its 3 nearest neighbors (n cells = 128,973, N spheroids = 6,236). dotted magenta line: first decile and dotted cyan line: last decile of the distances. f Corresponding histograms of all the nuclei and of the two gated populations of part e. (magenta=below the first decile; cyan=above the last decile). Dashed lines give the median values. g Confocal image of the mid-plane of a spheroid stained for phalloidin (red) and DAPI (blue). Dashed lines emphasize two representative cells. h Normalized ALB vs. distance from spheroid center (n ALB maxima = 87,609, N spheroids = 4,654). i Corresponding histogram for gated populations. j Confocal image of the mid-plane of a spheroid stained for ALB (green). White dashed line shows the spheroid edge. Non significant N.S., ***p < 0.001. a, b Kruskal–Wallis ANOVA followed by Mann–Whitney U-tests with Sidak’s correction for multiple comparisons. c Welch’s ANOVA followed by Games-Howell post hoc procedure
