Multiplexing high-content flow (HCF) and quantitative high-throughput screening (qHTS) to identify compounds capable of decreasing cell viability, activating caspase 3/7, expressing annexin V, and changing mitochondrial membrane integrity.
Multiplexing high-content flow (HCF) and quantitative high-throughput screening (qHTS) to identify compounds capable of decreasing cell viability, activating caspase 3/7, expressing annexin V, and changing mitochondrial membrane integrity.
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多重高内涵流 (HCF) 和定量高通量筛选 (qHTS) 来鉴定能够降低细胞活力、激活 caspase 3/7、表达膜联蛋白 V 和改变线粒体膜完整性的化合物。
DOI：
10.1002/9780470559277.ch130060
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发表时间：
2013
影响因子：
--
通讯作者：
Ferrer,Marc
中科院分区：
文献类型：
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作者：
Mathews,LesleyA;Keller,JonathanM;McKnight,Crystal;Michael,Sam;Shinn,Paul;Liu,Dongbo;Staudt,LouisM;Thomas,CraigJ;Ferrer,Marc
高含量流式(HCF)筛选系统，如INTELLICIT的IICE筛选系统和HTFC筛选系统，促进了用于高通量筛选的流式细胞仪分析的实施。HCF筛选系统能够使用较小的样本体积和多路分析来同时评估来自单个井的不同细胞参数。这在处理有限数量的电池或化合物时或在进行大规模筛选时变得无价。当分析可以小型化到384孔或1536孔的微孔板格式时，就有可能实现基于剂量响应的高通量筛选，也称为定量HTS或qHTS。本文描述了新的国家先进翻译科学中心(NCATS)的qHTS如何与HTFC筛查系统和Intelrict的多参数细胞凋亡筛查试剂盒系统地耦合，以使用弥漫性大B细胞淋巴瘤(DLBCL)模型对来自原代细胞增殖筛查的活性化合物进行生物学验证。普罗托克。化学。比奥尔。5：195-212©2013，John Wiley&Sons，Inc.
High‐content flow (HCF) screening systems, such as the iQue Screener and HTFC Screening System from IntelliCyt, have facilitated the implementation of flow cytometry assays for high‐throughput screening. HCF screening systems enable the use of smaller sample volumes and multiplexed assays to simultaneously assess different cellular parameters from a single well. This becomes invaluable when working with cells or compounds that are available in limited quantities or when conducting large‐scale screens. When assays can be miniaturized to a 384‐ or 1536‐well microplate format, it is possible to implement dose‐response‐based high‐throughput screens, also known as quantitative HTS or qHTS. This article describes how qHTS at the new National Center for Advancing Translational Science (NCATS) has been systematically coupled with the HTFC Screening System and Multimetric Apoptosis Screening Kit from IntelliCyt to biologically validate active compounds from primary cell proliferation screens using a model of diffuse large B cell lymphoma (DLBCL).Curr. Protoc. Chem. Biol. 5:195‐212 © 2013 by John Wiley & Sons, Inc.