Interaction between antigen-specific T cells and antigen presenting cells (APC) cognate ligand involve reorganization of the cytoskeleton and recruitment of adhesive and signaling molecules to the site of intercellular contact. Sustained adhesion of T cells to APCs and formation of the immunological synapse after T cell receptor stimulation are required for the antigen-specific response. One way to measure an immunological synapse is by fluorescently labeling the molecules that have been recruited to the synapse and imaging via confocal or conventional fluorescence microscopy. However, immunological synapses are often rare and therefore difficult to analyze objectively and statistically by traditional microscopy methods. In this study, imaging flow cytometry was employed to collect imagery of large numbers of cells to assess the percentage of T cells involved in an organized immunological synapse.
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