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Related Concept Videos

T Cell Activation and Clonal Selection01:22

T Cell Activation and Clonal Selection

548
T cells are integral to our adaptive immune system, recognizing and effectively responding to foreign antigens. T cell activation and clonal selection are pivotal in orchestrating this immune response. This article elucidates these mechanisms, detailing the roles of cluster of differentiation (CD) markers, major histocompatibility complex (MHC) molecules, costimulatory signals, and the process of clonal selection.
Naive T cells that have not yet encountered an antigen express two primary CD...
548

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Related Experiment Video

Updated: May 14, 2025

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Analyzing Point Accumulation in Nanoscale Topography (PAINT) Data to Decipher T Cell Receptor Cluster Signalling.

Shirin Ansari1, James C Walsh1, Jesse Goyette2

  • 1Department of Molecular Medicine, School of Biomedical Sciences, University of New South Wales, Sydney, NSW, Australia.

Methods in Molecular Biology (Clifton, N.J.)
|April 12, 2025
PubMed
Summary

This study introduces a new pipeline for analyzing PAINT data to quantify T cell receptor (TCR) nanoclusters. This method enhances the understanding of molecular events during T cell activation.

Keywords:
DNA-PAINTProtein-PAINTSuper-resolution microscopyT cell receptorTCRTCR nanocluster

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Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • T cell receptor (TCR) activation involves phosphorylation and clustering, initiating signaling cascades.
  • Studying molecular-level TCR activation requires high-resolution microscopy techniques.
  • Single-Molecule Localization Microscopy (SMLM) offers molecular precision for nanoscale events.

Purpose of the Study:

  • To present a novel pipeline for analyzing Points Accumulation In Nanoscale Topography (PAINT) data.
  • To quantify T cell receptor (TCR) nanoclusters in activated T cells using PAINT.
  • To leverage PAINT's advantages over traditional SMLM for accurate molecular quantification.

Main Methods:

  • Utilized Single-Molecule Localization Microscopy (SMLM) with PAINT techniques (DNA-PAINT, protein-PAINT).
  • Developed and applied a data analysis pipeline using the Picasso software package.
  • Collected and analyzed PAINT data from activated T cells.

Main Results:

  • Successfully quantified TCR nanoclusters in activated T cells.
  • Demonstrated the pipeline's capability to analyze PAINT data for molecular organization.
  • Highlighted PAINT's advantages in overcoming spectral limitations and photobleaching.

Conclusions:

  • The presented pipeline enables accurate quantification of TCR nanoclusters using PAINT.
  • This approach advances the study of molecular dynamics in T cell activation.
  • PAINT offers superior quantification for nanoscale biological events compared to conventional SMLM.