Flow cytometry and cell sorting
Flow cytometry and cell sorting
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DOI：
10.1007/10_2007_073
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发表时间：
2007-01-01
期刊：
影响因子：
--
通讯作者：
van den Engh, Ger
中科院分区：
文献类型：
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作者：
Ibrahim, Sherrif F.;van den Engh, Ger
流式细胞术和细胞分选技术在临床诊断和生物医学研究中是非常成熟的技术。细胞的异质混合物被放置在悬浮液中，并通过一个或多个激光询问点。从颗粒发出的光信号被收集起来，并与细胞形态、表面和细胞内蛋白表达、基因表达和细胞生理等实体相关联。根据用户定义的参数，单个细胞可以从流体流中分离出来，并以极快的速度和接近100%的纯度收集成可存活的均匀馏分。因此，细胞分选器成为众多下游研究的起点。流式细胞术是临床诊断的基石，而且更便宜、更通用的机器正在进入广泛和不同的用途。此外，计算和光学方面的进步已经导致新一代流式细胞仪能够以比其前辈快几个数量级的速度处理细胞，并且具有惊人的复杂性，使流式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。剩下的部分将分为临床和基于研究的流式细胞术和细胞分选的应用，重点介绍说明这种不可或缺的技术的多功能性的突出研究。
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.