Ashcroft R G, Lopez P A
Cytomation Inc., 4850 Innovation Drive, 80525, Fort Collins, CO, USA.
J Immunol Methods. 2000 Sep 21;243(1-2):13-24. doi: 10.1016/s0022-1759(00)00219-2.
Two recent events have opened a new domain of flow cytometry applications which we term high throughput flow cytometry (HTFC). The release of a commercial high speed sorter in 1994 placed HTFC within the reach of anyone who could buy one of the new machines and not just the handful of advanced laboratories worldwide that had custom built their own high speed sorters. The advent in 1999 of HTFC analysis capabilities of 100000 cells/s marks the second stage in this enabling of HTFC. We describe the technical basis of HTFC. The commercial high speed sorters measure cells in dead-times three to six times shorter than conventional machines. They can sort with high yield and high purity at rates from 25000 to 60000 cells/s, depending on their settings, mainly by virtue of their use of high drop creation rates 100000 drops/s or more. Finally, one series can analyse the measured cells at rates exceeding these sort-rates and at least six times faster than conventional sorters could. The performance of the systems made by the three manufacturers can be readily assessed for single laser systems. Comparison becomes difficult for multiple beam machines, due to requirements for multi-beam sampling for each cell and due to the demands of fluorescence compensation between signals from one laser and between signals from two or three lasers. Applications are described in the field of rare cell analysis and isolation as well as from sorting of abundant cell populations.
最近的两件事开启了流式细胞术应用的一个新领域，我们称之为高通量流式细胞术（HTFC）。1994年一款商用高速分选仪的推出，使任何能够购买一台新机器的人都能使用HTFC，而不仅仅是全球少数几个自行定制高速分选仪的先进实验室。1999年HTFC分析能力达到每秒100000个细胞标志着HTFC发展的第二阶段。我们描述了HTFC的技术基础。商用高速分选仪测量细胞的死时间比传统机器短三到六倍。它们能够以每秒25000到60000个细胞的速度进行高产率和高纯度分选，具体速度取决于其设置，这主要得益于它们采用了每秒100000滴或更高的高速液滴生成率。最后，一个系列能够以超过这些分选速度的速率分析所测量的细胞，并且比传统分选仪至少快六倍。对于单激光系统，三家制造商生产的系统性能很容易评估。对于多光束机器，由于每个细胞需要多光束采样以及来自一个激光的信号之间和来自两个或三个激光的信号之间的荧光补偿要求，比较变得困难。文中描述了HTFC在稀有细胞分析与分离以及丰富细胞群体分选领域的应用。
