Figure 3.

Visualization of large-scale reorganization of the outer membrane of Jurkat T cells on activating coated surfaces by mesh fitting to 3D localization data. (a) Meshes were fit to DHPSF whole-cell TCR localization data of three Jurkat T cells tagged with HaloTag-TMR. Cells were fixed in solution (left, 1149 localizations), 5 minutes (center, 2495 localizations), or 10 minutes (right, 2397 localisations) after being dropped onto an activating OKT3 coated surface. Large-scale reorganization of the outer membrane caused by immune-response triggering and the formation of an immunological synapse is visualized at different stages. Cells are colored axially by deviation of inter-protein distance from a complete spatial randomness model of a membrane-bound protein with yellow, which indicates no deviation from model system, and red/black, which indicates a reduction in peak inter-protein distance compared to model system. The flat grey surface represents the coverslip of the experiment. The cell fixed in solution (left) has been rendered away from the coverslip to emphasize the resting state. (b–c) Top-down and side-on views of (a). (d) Mean inter-protein distance deviation of each cell shown in (a) from a complete spatial randomness model of a membrane-bound protein.