Fig. 3: Ultra-high content imaging reveals the spatial composition of intestinal stroma.
From: Intestinal stroma guides monocyte differentiation to macrophages through GM-CSF
a Schematic overview of experimental design. b Gating strategy for identification of stroma, immune, and epithelial populations using ultra-high content MACSimaTM Imaging Platform; gated populations are backgated to the image; scale bars indicate 100 µm in images on the left and 50 µm in images presented in higher magnification in areas (a–d); color codes for the lines defining gated cellular units are shown in upper left corner and color codes for marker signal are shown in lower left corner. c Gated populations projected in UMAP space. d Absolute numbers (cells/mm2) of cell subsets quantified across intestinal layers in 14 biologically independent non-inflamed and inflamed samples from adult gut resections; lymphoid tissue was excluded from the analyses. e Composition of stromal cells in non-inflamed and inflamed samples across intestinal layers. f Quantification of distance from stromal subsets to different microanatomical landmarks in 14 (epithelium) to 42 (endothelium, lymphatic endothelium) biologically independent samples; only mucosa was included in the calculation of the distance to epithelium; scale bar indicates 100 µm. Mean with SE in (d) and (f); Mann–Whitney test was used to compare differences between non-inflamed and inflamed samples (two-tailed p value) (d), and Kruskal–Wallis with Dunn’s multiple comparisons test was used to comparing differences among stromal populations (f), ns not significant, nd not detected.
