Three-Dimensional Structured Illumination Microscopy (3D-SIM) to Dissect Signaling Cross-Talks in Motile T-Cells

Methods Mol Biol. 2019:1930:41-50. doi: 10.1007/978-1-4939-9036-8_6.

Abstract

Visualization of signal transduction events in T-cells has always been a challenge due to their miniscule size. Recent advancement in super-resolution microscopy techniques presents many new opportunities to navigate the spatial and temporal signaling cross-talks in motile T-cells. Here, we provide technical details, optimal conditions, and critical practical considerations that need to be taken into account during cell handling, sample preparation, and image acquisition of motile T-cells for performing three-dimensional structured illumination microscopy (3D-SIM).

Keywords: 3D-SIM; Immunostaining; Super-resolution microscopy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Movement*
  • Cell Tracking / methods*
  • Cells, Cultured
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Imaging, Three-Dimensional / methods*
  • Intercellular Adhesion Molecule-1 / metabolism
  • Lighting / methods*
  • Lymphocyte Function-Associated Antigen-1 / metabolism
  • Microscopy, Fluorescence / methods*
  • Signal Transduction
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology
  • T-Lymphocytes / physiology*

Substances

  • ICAM1 protein, human
  • Lymphocyte Function-Associated Antigen-1
  • Intercellular Adhesion Molecule-1