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- W4313354810 abstract "Abstract 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 (IS) after T cell receptor stimulation are required for the antigen-specific response. One way to measure an IS is by fluorescently labeling the molecules that have been recruited to the synapse and imaging via microscopy. However, IS are often rare and therefore difficult to analyze objectively and statistically by traditional microscopy methods. To overcome these problems, we employed the Amnis FlowSight imaging flow cytometry platform to objectively collect imagery of large numbers of cells to assess the percentage of T cells involved in an organized IS. In this study, Raji B cells were loaded with Staphylococcal enterotoxin B (SEB) to make APCs. The SEB-loaded APCs were incubated with human T cells to create T cell-APC conjugates. In addition, we investigated the inhibition of actin by cytochalasin D and how that affects the T cell-APC conjugates. The T cells, APCs and conjugates were fluorescently labeled. Using the FlowSight imaging flow cytometry platform we demonstrate image-based parameters that were used to assess the frequency of conjugates with an organized IS in an objective and statistically significant manner." @default.
- W4313354810 created "2023-01-06" @default.
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- W4313354810 date "2013-05-01" @default.
- W4313354810 modified "2023-09-25" @default.
- W4313354810 title "Measuring immunological synapse and actin organization using the FlowSight imaging flow cytometer (P5018)" @default.
- W4313354810 doi "https://doi.org/10.4049/jimmunol.190.supp.41.10" @default.
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