| Abstract | The plasma membrane is a complex and heterogeneous system, and critical biological processes take place in this environment. Microscopy methods are able to provide direct observations of membrane components in live cells. While microscopy is limited by its resolution, nanometer-scale interactions can still be observed either directly or indirectly. The diffusion of membrane components can be a useful parameter for determining the microscopic environment of specific objects. For example, different lipid compositions may have altered viscosity or large protein complexes may experience retarded diffusion. Our group is interested in the observation of T cell membrane proteins using single particle tracking (SPT). We have used this method to observe changes in cytoskeletal contacts of the T cell adhesion proteins LFA-1, CD2, and CD45. In each of these systems, SPT is able to reveal changes in cytoskeletal interactions of the proteins as a result of cellular activation or other perturbations. Our current labeling strategies and developments in methods of data analysis for these systems will be presented. |
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