Department of Immunology, Weizmann Institute of Science, Rehovot 76100, Israel.
Brief Funct Genomics. 2013 Mar;12(2):99-108. doi: 10.1093/bfgp/els061. Epub 2013 Jan 17.
Lymphocyte populations show a high level of phenotypic variability and are highly heterogeneous in their gene expression patterns. Studying this cell-to-cell variability, the processes which generate it and its implications for lymphocyte function can be advanced by live cell imaging combined with measurements of gene expression at the single-cell level. However, until recently such studies were limited due to the high motility of primary lymphocytes following their activation, their clustering that precludes single-cell analysis and the prolonged duration of relevant processes such as cell differentiation. In this review, we describe recent methodological advances, which enable single-cell studies of primary lymphocytes, and present some applications of these new techniques. We focus our discussion on microwell arrays. These arrays are typically comprised of thousands of small microwells in which primary lymphocytes can be trapped and imaged over long periods of time. This allows for quantitative evaluation of various cellular processes including cell proliferation, cell death, cytokine secretion and measurements of gene expression at the single-cell level. These advances pave the way for future studies of population variability, dynamic cell responses, stochasticity in gene expression and intercellular interactions between functional lymphocytes in controlled microenvironments.
淋巴细胞群体表现出高水平的表型可变性，其基因表达模式高度异质。通过活细胞成像结合单细胞水平的基因表达测量来研究这种细胞间的可变性、产生它的过程及其对淋巴细胞功能的影响，可以取得进展。然而，直到最近，由于初级淋巴细胞在激活后的高迁移率、阻止单细胞分析的聚类以及细胞分化等相关过程的持续时间较长，此类研究受到限制。在这篇综述中，我们描述了最近的方法学进展，这些进展使我们能够对原代淋巴细胞进行单细胞研究，并介绍了这些新技术的一些应用。我们将讨论重点放在微井阵列上。这些阵列通常由数千个小的微井组成，原代淋巴细胞可以在其中被捕获并长时间成像。这允许对各种细胞过程进行定量评估，包括细胞增殖、细胞死亡、细胞因子分泌和单细胞水平的基因表达测量。这些进展为未来在受控微环境中研究群体变异性、动态细胞反应、基因表达的随机性以及功能性淋巴细胞之间的细胞间相互作用铺平了道路。
