Cells of the adaptive immune system recognize pathogenic peptides through specialized receptors on their membranes. The engagement of these receptors with antigen leads to cell activation, which induces profound changes in the cell including cytoskeleton remodeling and membrane deformation. During this process, receptors and signaling molecules undergo spatiotemporal reorganization to form signaling microclusters and the immunological synapse. The cytoskeletal and membrane dynamics also leads to exertion of forces on the cell-substrate interface. In this chapter we describe two techniques-one for single-molecule imaging of B cell receptors to measure their diffusive properties as cells get activated on supported lipid bilayers; and the second for visualizing and quantifying cellular forces using elastic surfaces to stimulate T and B cells.
Keywords: B cell; B cell receptor; Cytoskeletal forces; Signaling; Single-molecule imaging; Substrate stiffness; Traction force microscopy.