A practical approach to multicolor flow cytometry for immunophenotyping
A practical approach to multicolor flow cytometry for immunophenotyping
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DOI：
10.1016/s0022-1759(00)00229-5
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发表时间：
2000-09-21
影响因子：
2.2
通讯作者：
Roederer, M
中科院分区：
文献类型：
--
作者：
Baumgarth, N;Roederer, M
通过流式细胞术硬件、软件和染料化学的一系列新发展，现在可以同时测量每个细胞上多达II种不同的荧光和两个散射光参数。这种先进的多色系统比目前的二色和三色流式细胞术测量有许多优点。它们为所研究的每个样本提供了大量的新信息，甚至对罕见的细胞群进行了精确的定量，并允许鉴定和表征新的细胞亚群。特别是，这项技术被证明对于在极其复杂的免疫系统中识别功能相同的细胞亚群至关重要；这种识别和枚举对于了解疾病的发病机制是重要的。然而，多色流式细胞术带来了一组新的，有时是困难的技术问题，用户必须克服这些问题才能获得有意义的结果。在这篇文章中，我们描述了多色流式细胞术的基本方面，包括可能出现的技术障碍和伪影，并就如何最好地克服这些障碍提供了一些建议。虽然受到我们目前使用的ii色技术的启发，但这些原则适用于使用多种荧光染料的所有流式细胞术实验。(C) 2000 Elsevier Science B.V.版权所有
Through a series of novel developments in flow cytometry hardware, software, and dye-chemistry it is now possible to simultaneously measure up to II distinct fluorescences and two scattered light parameters on each cell. Such advanced multicolor systems have a number of advantages over current two- and three-color flow cytometric measurements. They provide a large amount of novel information for each sample studied, an exquisitely accurate quantitation of even rare cell populations, and allow identification and characterization of novel cell subsets. In particular, this technology is proving crucial to identifying functionally homogeneous subsets of cells within the enormously complex immune system; such identification and enumeration is important for understanding disease pathogenesis. However, multicolor flow cytometry comes with a new and sometimes difficult set of technical problems that must be overcome by users to derive meaningful results. In this manuscript, we describe the basic aspects of multicolor flow cytometry, including the technical hurdles and artefacts that may occur, and provide some suggestions for how to best overcome these hurdles. While inspired by the Ii-color technology that we currently use, these principles apply to all flow cytometric experiments in which more than one fluorescent dye is used. (C) 2000 Elsevier Science B.V. All rights reserved.