Research Services Core (RSC)

研究服务核心(RSC)的工作人员为NCATS的高通量筛查、分析开发和优化方面的活动提供支持，并为NCATS筛查活动提供后续和剂量响应库电镀服务。化合物管理团队与自动化团队合作，为NCATS筛查活动提供后续和剂量响应库电镀服务。自动化团队支持的系统包括初级筛查系统、Tox21筛查系统、RNAi筛查系统、用于分析后续工作的RNAi Hit拾取系统以及高分辨率生物系统(HRB)声学电镀系统。在过去的一年里，我们还安装并开始运行Thermo Science Spinnaker平板制备工作站，用于将化合物以批量形式固定并将试剂分配到平板中。所有这些系统结合在一起，提供了广泛的自动化和复合电镀任务，用于实现NCATS内部科学计划的所有方面。 初步筛选系统使用三个用于平板运输的机械臂、300多万个复合孔(约2,000个1,536孔复合板)和150多万个化验井(约1,100个1,536孔分析板)的存储能力，以实现世界级的筛选生产率，同时保持研究人员所需的高水平数据质量。它还具有并行运行多个检测的能力。中心专家开发了定制软件来监视和控制系统，此外还实时存档了每个屏幕的完整过程历史。Tox21筛选系统使用一个用于平板运输的机械臂和近100万个化合物或化验孔(约600个1,536孔化合物或化验板)的存储能力来筛选初级筛选系统和Tox21化合物库中没有的较小规模的化合物文库。该平台旨在补充主要系统，并且可以轻松定制，允许快速集成其他筛查技术。 RSC团队还为NCATS的内部项目提供筛查系统服务。Tox21筛选系统包含一个用于平板运输的机械臂和约150万个复合孔(约1,000个1,536孔复合板)和150多万个化验井(约1,100个1,536孔分析板)的存储能力。通过一个销钉工具和两个用于化合物添加的声学分配器、四个用于试剂添加的小容量分配器和三个平板阅读器，该系统可以快速筛选包含10,000种化合物的21世纪毒理学(Tox21)文库，一式三份。今年，RSC团队还完成了Hamamatsu FDSS平板阅读器的安装，使Tox21程序能够扩展平台上运行的各种分析，特别是钙通道。FDSS本身被修改为包括由Ionfield开发的定制1536针式工具清洁剂；这是第一次使用冷等离子技术而不是DMSO和其他溶剂进行清洁，既提供了更好的数据，因为最大限度地减少了板之间的结转影响，并减少了对腐蚀性溶剂的需求，而腐蚀性溶剂在过去曾损坏仪器。NCATS专家使用与初级筛查系统相同的定制软件来监测和控制该系统。RNAi筛选系统有两个用于平板运输的机械臂，以及用于检测平板存储的孵化器和促进系统连续操作的平板堆叠器。多通道移液器、低容量分配器、平板洗涤吸入器和两种不同的平板读取器支持这个RNAi聚焦系统的各种检测方法。 RSC工作人员还继续支持新成立的干细胞翻译实验室(SCTL)。不同的团队成员提供了大量的支持，提供了大量的设备，包括自动组织培养系统、流式细胞仪、细胞分选仪和高含量成像仪。特别值得一提的是，自动化组织培养平台已经成为所有NCATS研究人员的核心能力，需求变得如此之大，以至于很可能会有另一个系统订购。这些资源需要大量的专业知识来操作和维护。在过去的一年里，RSC已经开始成为这些技术的核心设施，使不同的生物小组能够提高他们的能力。与高含量成像仪有关，过去一年进行了更多的高含量分析，这是因为与前四年的总和相比，提高了能力。

Research Services Core (RSC) staff provide support to the NCATS activities in high-throughput screening, assay development, and optimization as well as follow-up and dose-response library-plating services for NCATS screening activities. The Compound Management team works in conjunction with the Automation team to provide follow-up and dose-response library-plating services for NCATS screening activities. Systems supported by the Automation team include primary screening system, Tox21 screening system, RNAi screening system, the RNAi hit picking system for assay follow ups, and the High Resolution Biosystems (HRB) acoustic plating system. In the past year, we have also installed and begun to operate a Thermo Scientific Spinnaker plate preparation workstation used to pin compounds and dispense reagents into plates in a batch format. Together, all of these systems combined provide a wide variety of automation and compound plating tasks used to enable all facets of the NCATS intramural scientific program. The primary screening system utilizes three robotic arms for plate transportation, a storage capacity of more than 3 million compound wells (approximately 2,000 1,536-well compound plates), and more than 1.5 million assay wells (approximately 1,100 1,536-well assay plates) to achieve a world-class screening productivity while maintaining the high level of data quality required by researchers. It also has the ability to run multiple assays in parallel. Center experts have developed custom software to monitor and control the system, in addition to archiving the complete process history for every screen in real time. The Tox21 screening system uses a single robotic arm for plate transportation and a storage capacity of nearly 1 million compound or assay wells (about 600 1,536-well compound or assay plates) to screen smaller scale compound libraries not present on the primary screening system and the Tox21 compound library. This platform is designed to complement the primary system, and it is easily customizable, allowing for the rapid integration of additional screening technologies. The RSC team additionally provides screening system services to intramural programs within NCATS. The Tox21 screening system contains a single robotic arm for plate transportation and a storage capacity of about 1.5 million compound wells (about 1,000 1,536-well compound plates) and more than 1.5 million assay wells (approximately 1,100 1,536-well assay plates). With a pin tool and two acoustic dispensers for compound addition, four low-volume dispensers for reagent addition, and three plate readers enabling a variety of assay detection methods, this system can rapidly screen, in triplicate, the Toxicology in the 21st Century (Tox21) library of 10,000 compounds. This year, the RSC team was also able to complete the installation of the Hamamatsu FDSS plate reader allowing the Tox21 program to expand the kinds of assays run on the platform, in particular calcium channel. The FDSS itself was modified to include a custom 1536 pintool cleaner developed by IonField; which is a first of its kind using cold plasma technology instead of DMSO and other solvents to clean, providing both better data due to minimizing carry-over effects between plates and reducing the need for caustic solvents which in the past have damaged the instrument. NCATS experts monitor and control this system with the same custom software used for the primary screening system. The RNAi screening system has two robotic arms for plate transportation with incubators for assay plate storage and plate stackers to facilitate continuous system operation. A multichannel pipettor, low volume dispensers, an aspirator for plate washing, and two different plate readers support a wide variety of detection methods for this RNAi-focused system. RSC staff also continue to support the newly formed Stem Cell Translation Laboratory (SCTL). Various team members have provided a large amount of support with a vast array of equipment including automated tissue culture systems, flow cytometers, cell sorters, and high content imagers. Special mention should be given to the automated tissue culture platform which has established itself as a core capability for all NCATS researchers now with the demand becoming so great that another system is most likely on order. These resources require a tremendous amount of expertise to operate and maintain. In the past year, RSC has begun to act as a core facility for these technologies, enabling different biology groups to increase their capabilities. Related to the high content imagers, more high content assays were run in the past year due to the increased capabilities as compared to the previous four years combined.