Figure 2
From: Controlling T cells spreading, mechanics and activation by micropatterning
(a) RICM images of adhesion and spreading of 3A9m cells on anti CD45 “continuous” (here denominated as “cont.”) substrate at given time points. (b) Adhesion area on anti CD45 antibodies of cells vs. time at 25 °C and 37 °C. Each point represents a cell, and the bar denotes the median value. Statistical differences between temperatures at each time point were assessed using Mann–Whitney test. The adhesion after 60 min at 37 °C corresponds to an equivalent disk of diameter of ~ 20 µm. (c) Schematics of the inverse microstamping method, where the repulsive molecule (BSA or bBSA) is stamped first, before adsorbing the adhesive one (anti CD45) in the free zones. (d) Images of obtained substrates: (i) fluorescence image of the repulsive zones, completed with non labeled anti CD45 antibodies in the patterns (dark), (ii) brightfield images of 3A9m cells after 60 min incubation, (iii) corresponding RICM image of the adhesion zones, (iv) overlay of the three previous pictures. (e) Confocal imaging of 3A9m cells showing patterns of anti CD45 antibodies (green) cortical actin (red) and nucleus (blue), with projections of the z-stack. (f) Estimation of the distance between the cortical actin and the nucleus (mean+/SEM), to determine the maximal indentation for the Hertz-like fit (dotted line, see text). Scale bar = 20 µm.
