Quantification of T-Cell Migratory Phenotypes Using High-Content Analysis

Methods Mol Biol. 2019:1930:25-32. doi: 10.1007/978-1-4939-9036-8_4.

Abstract

The exploration screening of phenotypic changes in motile T-cells within a signaling environment has always been an arduous task due to the sheer population of these microscopic cells. In recent years, High-Content Analysis (HCA) has gained epochal momentum and has allowed for a wider range of quantitative multiplexed cell-based assays in the field of lymphocyte signaling. In this chapter, we consolidate our understanding and describe the technical approach and methodology to quantify T-cell migratory phenotypes using HCA. Optimizations to be adopted to generate high-quality cytological images of motile T-cells and subsequent analysis using HCA are detailed as well.

Keywords: High-content analysis; Immunostaining; T-cell migration.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Movement*
  • Cells, Cultured
  • High-Throughput Screening Assays / methods*
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Intercellular Adhesion Molecule-1 / metabolism
  • Lymphocyte Function-Associated Antigen-1 / metabolism
  • Microscopy, Fluorescence / methods*
  • Phenotype
  • Signal Transduction
  • T-Lymphocytes / cytology
  • T-Lymphocytes / metabolism
  • T-Lymphocytes / physiology*

Substances

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