Ibrahim Sherrif F, van den Engh Ger
Department of Dermatology, University of Rochester, 601 Elmwood Avenue, Rochester, NY 14642, USA.
Adv Biochem Eng Biotechnol. 2007;106:19-39. doi: 10.1007/10_2007_073.
Flow cytometry and cell sorting are well-established technologies in clinical diagnostics and biomedical research. Heterogeneous mixtures of cells are placed in suspension and passed single file across one or more laser interrogation points. Light signals emitted from the particles are collected and correlated to entities such as cell morphology, surface and intracellular protein expression, gene expression, and cellular physiology. Based on user-defined parameters, individual cells can then be diverted from the fluid stream and collected into viable, homogeneous fractions at exceptionally high speeds and a purity that approaches 100%. As such, the cell sorter becomes the launching point for numerous downstream studies. Flow cytometry is a cornerstone in clinical diagnostics, and cheaper, more versatile machines are finding their way into widespread and varied uses. In addition, advances in computing and optics have led to a new generation of flow cytometers capable of processing cells at orders of magnitudes faster than their predecessors, and with staggering degrees of complexity, making the cytometer a powerful discovery tool in biotechnology. This chapter will begin with a discussion of basic principles of flow cytometry and cell sorting, including a technical description of factors that contribute to the performance of these instruments. The remaining sections will then be divided into clinical- and research-based applications of flow cytometry and cell sorting, highlighting salient studies that illustrate the versatility of this indispensable technology.
流式细胞术和细胞分选是临床诊断和生物医学研究中成熟的技术。将细胞的异质混合物置于悬浮液中，并单个通过一个或多个激光检测点。从颗粒发出的光信号被收集起来，并与细胞形态、表面和细胞内蛋白质表达、基因表达以及细胞生理学等实体相关联。根据用户定义的参数，然后可以将单个细胞从流体流中分离出来，并以极高的速度和接近100%的纯度收集到有活力的、同质的组分中。因此，细胞分选仪成为众多下游研究的起点。流式细胞术是临床诊断的基石，更便宜、用途更广泛的仪器正被广泛应用于各种不同的用途。此外，计算和光学方面的进步催生了新一代流式细胞仪，其处理细胞的速度比前代产品快几个数量级，且复杂度惊人，使细胞仪成为生物技术中强大的发现工具。本章将首先讨论流式细胞术和细胞分选的基本原理，包括对影响这些仪器性能的因素的技术描述。其余部分将分为基于临床和研究的流式细胞术和细胞分选应用，重点介绍一些突出的研究，以说明这项不可或缺的技术的多功能性。
