Fig. 6.

Putative stromal macrophages are morphologically and behaviorally distinct from resident tissue keratocytes and are affected by allergy and acute stimulation elicited by CL wear. (A) Sequential time-lapsed images of the anterior stroma in a healthy control cornea over a 20-min period. Stationary keratocytes (pink arrows) and motile cells (orange arrowheads) demonstrate differential behaviors. A color-coded hyperstack shows cytoplasmic displacement in macrophages, and cumulative cell masks show small-shaped keratocytes (magenta) compared to ameboid-shaped macrophages (cyan). (B) The coefficient of variation of cell shape over time was higher in macrophages compared to keratocytes. (C) Keratocytes represent the major cell type in the anterior stroma, followed by macrophages and T cells. (D) There was a similar density of stromal macrophages in individuals with untreated and treated allergy compared to healthy controls. Per cell analysis plots of cell area (E), perimeter (F), and circularity (G) show differences in the morphology of stromal macrophages in control vs. treated and untreated allergy. (H) Representative time-lapse color-coded hyperstacks of stromal macrophages in healthy control, untreated allergy, and treated allergy individuals. (I) Time-lapse image sequence and corresponding color-coded hyperstacks and cumulative cell area masks of a stromal macrophage pre-CL (Top row) and post-CL (Bottom row). (J) Repeated measures comparisons of the area, perimeter, solidity, and circularity of the same stromal macrophages (n = 9) were quantified pre-CL and post-CL. Scale bars are as indicated. Data are plotted as mean ± SD. *P < 0.05; **P < 0.01, ***P < 0.001, ****P < 0.0001.