Ng Aik Seng, Ong Seow Theng, Kelleher Dermot, Verma Navin Kumar
Lymphocyte Signalling Research Laboratory, Lee Kong Chian School of Medicine, Nanyang Technological University Singapore, Singapore, Singapore.
School of Biological Sciences, Nanyang Technological University Singapore, Singapore, Singapore.
Methods Mol Biol. 2019;1930:25-32. doi: 10.1007/978-1-4939-9036-8_4.
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.
由于这些微小细胞数量众多,在信号环境中对运动性T细胞的表型变化进行探索性筛选一直是一项艰巨的任务。近年来,高内涵分析(HCA)获得了划时代的发展动力,并在淋巴细胞信号传导领域实现了更广泛的基于细胞的定量多重分析。在本章中,我们总结了我们的理解,并描述了使用HCA量化T细胞迁移表型的技术方法。还详细介绍了为生成运动性T细胞的高质量细胞学图像以及随后使用HCA进行分析而应采用的优化方法。