Quantification of T-Cell Migratory Phenotypes Using High-Content Analysis
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 wordsHigh-content analysis Immunostaining T-cell migration
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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