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MIRAVA POLYSCOPE – All in one and on for all: the perfect image
Science beyond Barriers

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Technology

2023
The Immune Synapse: Methods and Protocols

A DNA Origami-Based Biointerface to Interrogate the Spatial Requirements for Sensitized T-Cell Antigen Recognition

Authors:

Joschka Hellmeier, René Platzer, Johannes B Huppa, Eva Sevcsik

Keywords:

DNA origami, T cell, Antigen recognition, Single molecule, Fluorescence microscopy, Supported lipid bilayer

Abstract:

When T cells scan the surface of antigen-presenting cells (APCs), they can detect the presence of just a few antigenic peptide/MHC complexes (pMHCs), in some cases even a single agonist pMHC. These are typically vastly outnumbered by structurally similar yet non-stimulatory endogenous pMHCs. How T cells achieve this enormous sensitivity and selectivity is still not clear, in particular in view of the rather moderate (1-100 μM) affinity that T-cell receptors (TCRs) typically exert for antigenic pMHCs. Experimental approaches that enable the control and quantification of physical input parameters within the context of the immunological synapse to precisely interrogate the molecular consequences of TCR-engagement, appear highly advantageous when searching for better answers.We here describe the implementation of a biointerface that allows to experimentally define molecular distances between T-cell ligands as a means to correlate them with molecular dynamics of antigen engagement, downstream signaling, and the overall T-cell response. The basis of this biointerface is DNA origami nanostructures, which are (i) rigid and highly versatile platforms that can (ii) be embedded as laterally mobile entities within supported lipid bilayers and functionalized (iii) in a site-specific and orthogonal manner with (iv) one or more proteins of choice.

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Superresolution & Confocal Systems

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Superresolution & Confocal Modules

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  • MINFLUX Module
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