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| Item Type: | Article |
|---|---|
| Title: | Three-dimensional single molecule localization microscopy reveals the topography of the immunological synapse at isotropic precision below 15 nm |
| Creators Name: | Velas, L., Brameshuber, M., Huppa, J.B., Kurz, E., Dustin, M.L., Zelger, P., Jesacher, A. and Schütz, G.J. |
| Abstract: | T-cells engage with antigen-presenting cells in search for antigenic peptides and form transient interfaces termed immunological synapses. Synapse topography affects receptor binding rates and the mutual segregation of proteins due to size exclusion effects. It is hence important to determine the 3D topography of the immunological synapse at high precision. Current methods provide only rather coarse images of the protein distribution within the synapse. Here, we applied supercritical angle fluorescence microscopy combined with defocused imaging, which allows three-dimensional single molecule localization microscopy (3D-SMLM) at an isotropic localization precision below 15 nm. Experiments were performed on hybrid synapses between primary T-cells and functionalized glass-supported lipid bilayers. We used 3D-SMLM to quantify the cleft size within the synapse by mapping the position of the T-cell receptor (TCR) with respect to the supported lipid bilayer, yielding average distances of 18 nm up to 31 nm for activating and nonactivating bilayers, respectively. |
| Keywords: | T-Cells, Immunological Synapse, Interference Reflection Microscopy, T-Cell Receptor, 3D Single Molecule Localization Microscopy |
| Source: | Nano letters |
| ISSN: | 1530-6992 |
| Publisher: | American Chemical Society (ACS) |
| Volume: | 21 |
| Number: | 21 |
| Page Range: | 9247-9255 |
| Date: | 10 November 2021 |
| Official Publication: | https://doi.org/10.1021/acs.nanolett.1c03160 |
| PubMed: | View item in PubMed |
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