Black Christopher B, Duensing Thomas D, Trinkle Linda S, Dunlay R Terry
IntelliCyt Corporation, 317 Commercial St. NE, Albuquerque, NM 87102, USA.
Assay Drug Dev Technol. 2011 Feb;9(1):13-20. doi: 10.1089/adt.2010.0308. Epub 2010 Nov 4.
This review describes the use of high-throughput flow cytometry for performing multiplexed cell-based and bead-based screens. With the many advances in cell-based analysis and screening, flow cytometry has historically been underutilized as a screening tool largely due to the limitations in handling large numbers of samples. However, there has been a resurgence in the use of flow cytometry due to a combination of innovations around instrumentation and a growing need for cell-based and bead-based applications. The HTFC™ Screening System (IntelliCyt Corporation, Albuquerque, NM) is a novel flow cytometry-based screening platform that incorporates a fast sample-loading technology, HyperCyt®, with a two-laser, six-parameter flow cytometer and powerful data analysis capabilities. The system is capable of running multiplexed screening assays at speeds of up to 40 wells per minute, enabling the processing of a 96- and 384-well plates in as little as 3 and 12 min, respectively. Embedded in the system is HyperView®, a data analysis software package that allows rapid identification of hits from multiplexed high-throughput flow cytometry screening campaigns. In addition, the software is incorporated into a server-based data management platform that enables seamless data accessibility and collaboration across multiple sites. High-throughput flow cytometry using the HyperCyt technology has been applied to numerous assay areas and screening campaigns, including efflux transporters, whole cell and receptor binding assays, functional G-protein-coupled receptor screening, in vitro toxicology, and antibody screening.
本综述介绍了高通量流式细胞术在基于细胞和基于微珠的多重筛选中的应用。随着基于细胞的分析和筛选技术的诸多进展,流式细胞术在历史上一直未得到充分利用,主要是因为在处理大量样本方面存在局限性。然而,由于仪器方面的创新以及对基于细胞和基于微珠应用的需求不断增加,流式细胞术的使用再度兴起。HTFC™筛选系统(IntelliCyt公司,新墨西哥州阿尔伯克基)是一种基于流式细胞术的新型筛选平台,它将快速样本加载技术HyperCyt®与双激光、六参数流式细胞仪及强大的数据分析功能相结合。该系统能够以每分钟高达40孔的速度进行多重筛选测定,分别只需3分钟和12分钟就能处理96孔板和384孔板。系统中嵌入了HyperView®,这是一个数据分析软件包,可从多重高通量流式细胞术筛选活动中快速识别出命中结果。此外,该软件被集成到一个基于服务器的数据管理平台中,可实现多个站点之间的数据无缝访问与协作。使用HyperCyt技术的高通量流式细胞术已应用于众多检测领域和筛选活动,包括外排转运体、全细胞和受体结合测定、功能性G蛋白偶联受体筛选、体外毒理学以及抗体筛选。