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

  • Aik Seng Ng
  • Seow Theng OngEmail author
  • Dermot Kelleher
  • Navin Kumar Verma
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1930)

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.

Key words

High-content analysis Immunostaining T-cell migration 

Notes

Acknowledgments

This work was supported by Lee Kong Chian School of Medicine, Nanyang Technological University Singapore Start-Up Grant and the Ministry of Education (MOE), Singapore under its Singapore MOE Academic Research Fund (AcRF) Tier 1 (2014-T1-001-141) and MOE-AcRF Tier 2 (MOE2017-T2-2-004) grants to N.K.V. The authors would like to thank Jaron Liu (GE Healthcare) for his expert advice on the functionality of the equipment. AS Ng would also like to acknowledge the funding support for this project from Nanyang Technological University under the Undergraduate Research Experience on CAmpus (URECA) programme.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Aik Seng Ng
    • 1
    • 2
  • Seow Theng Ong
    • 1
    Email author
  • Dermot Kelleher
    • 1
    • 3
  • Navin Kumar Verma
    • 4
  1. 1.Lymphocyte Signalling Research Laboratory, Lee Kong Chian School of MedicineNanyang Technological University SingaporeSingaporeSingapore
  2. 2.School of Biological SciencesNanyang Technological University SingaporeSingaporeSingapore
  3. 3.Department of Medicine and Biochemistry and Molecular BiologyUniversity of British ColumbiaVancouverCanada
  4. 4.Lee Kong Chian School of MedicineNanyang Technological University SingaporeSingaporeSingapore

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