Tcr Micro Clusters
Tcr Micro Clusters - Related Videos
19:05
Measuring TCR-pMHC Binding In Situ using a FRET-based Microscopy Assay
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Cited by 9 •
2015
This manuscript describes how to conduct (single molecule) Förster Resonance Energy Transfer (FRET)- based assays to measure the binding dynamics between T-cell antigen receptor (TCR) and antigenic peptide-loaded MHC molecules as they occur within the immunological synapse of a T-cell in contact with a functionalized planar supported lipid bilayer.
16:10
A TIRF Microscopy Technique for Real-time, Simultaneous Imaging of the TCR and its Associated Signaling Proteins
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Cited by 13 •
2012
The compartmentalization of proteins either within the plasma membrane or into intracellular locations is one regulatory mechanism that can greatly influence signaling outcomes; hence, to understand signaling it is important to study the spatial and temporal behavior of the proteins involved. We describe here a TIRF microscopy based system to study signal transduction in T cells, but is broadly applicable.
11:21
Streamlined Single Cell TCR Isolation and Generation of Retroviral Vectors for In Vitro and In Vivo Expression of Human TCRs
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Cited by 3 •
2017
The current protocol combines single cell paired human TCR alpha and beta chain sequencing with streamlined generation of retroviral vectors compatible with in vitro and in vivo TCR expression.
06:42
Identifying Kinase Inhibitors that Modulate the Thymocyte Response to Strong TCR Signals
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2025
In this video, we describe a method to identify the small-molecule kinase inhibitors that modulate the apoptosis of self-reactive CD4+CD8+ double-positive immature thymocytes. Apoptosis is induced in the double-positive thymocytes by activating them with anti-CD3- and anti-CD28-coated magnetic beads; this is followed by a small-molecule inhibitor treatment and flow cytometry analysis to detect if the inhibitors modulate the apoptotic marker expression.
10:37
Spatial Separation of Molecular Conformers and Clusters
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Cited by 3 •
2014
We present a technique that allows the spatial separation of different conformers or clusters present in a molecular beam. An electrostatic deflector is used to separate species by their mass-to-dipole moment ratio, leading to the production of gas-phase ensembles of a single conformer or cluster stoichiometry.
FAQs
Peptide–MHC binding initiates the gathering of TCRs with CD3, ZAP-70, and LAT. Their close association creates a localized signaling platform in which phosphorylation events integrate receptor-derived information. This organization links antigen recognition to cytoskeletal remodeling and helps explain how an initially local molecular event can promote broader T-cell activation during immune responses.
CD3, ZAP-70, and LAT are important because they participate in the molecular assembly that follows TCR engagement. Within the cluster, their association supports phosphorylation-driven signal transmission rather than leaving receptor binding as an isolated event. Examining these components helps investigators connect cluster composition with the efficiency and coordination of downstream T-cell activation.
Tracking formation, movement, and dissolution shows that signaling organization changes over time rather than remaining fixed. These measurements can indicate when receptor-associated signaling assemblies appear, how they are repositioned, and when they disperse. Comparing these stages helps researchers relate the lifetime and behavior of a cluster to the progression or regulation of T-cell signaling.
Fluorescence microscopy can visualize molecular components, while live-cell imaging follows cluster behavior as cells respond over time. Researchers can use these approaches to examine when assemblies form and where they move, then apply quantitative analysis to compare their composition or dynamics. Together, the methods provide complementary spatial and temporal information about early T-cell signaling.
Quantitative analysis turns visual observations into measurements of formation, movement, composition, and dissolution. Rather than relying only on whether clusters are visible, investigators can compare patterns across experimental conditions or cell responses. This supports more precise interpretation of signal organization and can reveal changes in immune regulation or pathogen-specific T-cell responses.
In infection research, these assemblies offer a way to examine how pathogen-specific antigen recognition becomes coordinated T-cell activation. In broader immunology, their composition and dynamics can be evaluated when signaling is altered. Such studies may clarify mechanisms of immune regulation and provide cellular context for disorders involving abnormal T-cell signaling.