Osborne Geoffrey W
Queensland Brain Institute/Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, Australia.
Methods Cell Biol. 2011;102:533-56. doi: 10.1016/B978-0-12-374912-3.00021-3.
The classification and separation of one cell type or particle from others is a fundamental task in many areas of science. Numerous techniques are available to perform this task; however, electrostatic cell sorting has gained eminence over others because, when combined with the analysis capabilities of flow cytometry it provides flexible separations based on multiple parameters. Unlike competing technologies, such as gradient or magnetic separations that offer much larger total throughput, flow cytometric cell sorting permits selections based on various levels of fluorescent reporters, rather the complete presence or absence of the reporter. As such, this technology has found application in a huge range of fields. This chapter aims to describe the utility of single-cell sorting with particular emphasis given to index sorting. This is followed by two recently developed novel techniques of sorting cells or particles. The first of these is positional sorting which is useful in cell-based studies where sorting can proceed and produce meaningful results without being inherently dependant on prior knowledge of where gates should be set. Secondly, reflective plate sorting is introduced which positionally links multiwell sample and collection plates in a convenient assay format so that cells in the collection plate "reflect" those in the sample plate.
从其他细胞类型或颗粒中分类和分离出一种细胞类型是许多科学领域的一项基本任务。有许多技术可用于执行此任务；然而，静电细胞分选比其他技术更具优势，因为当与流式细胞术的分析能力相结合时，它可以基于多个参数进行灵活的分离。与诸如梯度分离或磁分离等竞争技术不同，后者可提供更大的总通量，流式细胞术细胞分选允许基于各种水平的荧光报告分子进行分选，而不是基于报告分子的完全存在或不存在。因此，这项技术已在众多领域得到应用。本章旨在描述单细胞分选的效用，特别强调索引分选。接下来是两种最近开发的分选细胞或颗粒的新技术。第一种是位置分选，它在基于细胞的研究中很有用，在这种研究中，分选可以进行并产生有意义的结果，而不必固有地依赖于关于应设置门控位置的先验知识。其次，介绍了反射板分选，它以方便的检测形式将多孔样品板和收集板在位置上联系起来，以便收集板中的细胞“反映”样品板中的细胞。
