Research Services Core (RSC)

研究服务核心 (RSC)

• 批准号: 10261247
• 负责人: Samuel Michael
• 金额: $ 1009.75万
• 依托单位: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10261247
• 关键词: 2019-nCoV; Acoustics; Adult; Affect; Antibodies; Aptitude; Archives; Automation; Award; Biological; Biological Assay; Biology; Blood; Blood specimen; COVID-19; Calcium Channel; Caustics; Cells; Chemical Structure; Chemicals; Clinical Trials; Collection; Communities; Complement; Complex; Computer software; Contracts; Core Facility; Custom; Data; Data Analyses; Databases; Detection; Development; Diagnostic; Dimethyl Sulfoxide; Dose; Emerging Technologies; Engineering; Enzyme-Linked Immunosorbent Assay; Equipment; Hybrids; Incubators; Infection; Institutes; Internet; Intramural Research Program; Laboratories; Libraries; Liquid substance; Maintenance; Manuals; Maryland; Methods; Mission; Monitor; National Institute of Allergy and Infectious Disease; National Institute of Biomedical Imaging and Bioengineering; Nature; Online Systems; Patients; Phase; Plasma; Population; Preparation; Process; Productivity; Protocols documentation; RNA Interference; RNA interference screen; Reader; Reagent; Recording of previous events; Research; Research Institute; Research Personnel; Research Project Grants; Research Support; Resolution; Resources; Retrieval; Robotics; Run-On Assays; Running; Sampling; Scientist; Services; Solvents; Support System; Surveys; System; Technology; Testing; Time; Toxicology; Training; Translations; Transportation; United States; United States National Institutes of Health; Update; Vial device; Virus; Work; arm; assay development; biosafety level 3 facility; cohesion; dashboard; data quality; design; epidemiologic data; epidemiological model; experimental study; follow-up; high throughput screening; imager; insight; instrument; instrumentation; member; new technology; next generation; novel coronavirus; operation; programs; recruit; response; screening; small molecule; software systems; stem cells; tissue culture; tool; vigilance; volunteer; web site

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 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. The NCATS intramural program is heavily involved in COVID-19 relief efforts across the NIH. RSC is directly involved in several of these key initiatives, providing project management, scientific and technical support across several programs. Below are a list of some of the COVID-19 projects RSC has worked on: NIH Serosurvey: A new study has begun recruiting at the National Institutes of Health in Bethesda, Maryland, to determine how many adults in the United States without a confirmed history of infection with SARS-CoV-2, the virus that causes coronavirus disease 2019 (COVID-19), have antibodies to the virus. The presence of antibodies in the blood indicates a prior infection. In this serosurvey, researchers will collect and analyze blood samples from as many as 10,000 volunteers to provide critical data for epidemiological models. The results will help illuminate the extent to which the novel coronavirus has spread undetected in the United States and provide insights into which communities and populations are most affected. NCATS and RSC are heavily involved in this effort on two fronts. For the survey itself, RSC is creating a web-based dashboard which will directly connect to the REDCap clinical trial software system being setup at NIAID. As results from the study are entered into REDCap, RSC is creating a dashboard which will programmatically pull the most recent results and update on a daily basis for program staff to monitor progress of the study and to make sure key milestones are being met. To analyze the samples provided by the volunteers, RSC is also assisting by providing automation and instrumentation expertise to the lab at NIBIB where the results are being processed. The diagnostic being run is an ELISA assay, with the results for patients being read out on a BMG Labtech PHERAstar plate reader. These results will also be updated to the same web-based dashboard so program staff can see this data as well. Although this initial phase is starting with 10,000 volunteers, the demand has been massive, and it is highly likely RSC will be involved with turning this currently manual process to run the ELISA assay into a fully automated process. Compound testing in the SARS-CoV-2 CPE assay (non-GLP): This project involves sending pre-spotted assay plates prepared by the ITRB Compound Management group to run assays at the Southern Research Institute (SRI). This contract was awarded (75N95020P00144) and five rounds of assays have already been completed. This data will be made publicly available via a website that RSC will provide all hosting support for called the OpenData Portal. In addition, the next round of experiments will be to take the hits from the initial experiment and to run them in a combination dose response fashion, referred to as a matrix screen. All this plating will be performed by RSC staff and it will take place on our LabCyte Access system. As more experiments are run, more data will be made available on the COVID-19 data website and more compounds will need to be sent to SRI and potentially other BSL3 facilities.

研究服务核心 (RSC) 工作人员为 NCATS 活动提供高通量筛选、测定开发和优化的支持，以及 NCATS 筛选活动的后续和剂量反应文库电镀服务。化合物管理团队与自动化团队合作，为 NCATS 筛选活动提供后续和剂量反应库电镀服务。自动化团队支持的系统包括初级筛选系统、Tox21 筛选系统、RNAi 筛选系统、用于后续分析的 RNAi 命中挑选系统以及高分辨率生物系统 (HRB) 声学电镀系统。去年，我们还安装并开始运行 Thermo Scientific Spinnaker 板制备工作站，用于将化合物固定并将试剂以批量形式分配到板中。 所有这些系统结合起来提供了各种自动化和复合电镀任务，用于实现 NCATS 校内科学计划的各个方面。 初级筛选系统利用三个机械臂进行板运输，存储容量超过300万个复合孔（约2,000个1,536孔复合板）和超过150万个分析孔（约1,100个1,536孔分析板），以实现世界一流的筛选生产力，同时保持研究人员所需的高水平数据质量。它还能够并行运行多个测定。除了实时存档每个屏幕的完整过程历史记录外，中心专家还开发了定制软件来监视和控制系统。 Tox21 筛选系统使用单个机械臂进行板运输，并具有近 100 万个化合物或分析孔（约 600 个 1,536 孔化合物或分析板）的存储容量，以筛选初级筛选系统和 Tox21 化合物库中不存在的较小规模的化合物库。该平台旨在补充主系统，并且可以轻松定制，从而可以快速集成其他筛选技术。 RSC 团队还为 NCATS 内的校内项目提供筛查系统服务。 Tox21 筛选系统包含一个用于板运输的机械臂，存储容量为约 150 万个化合物孔（约 1,000 个 1,536 孔化合物板）和超过 150 万个分析孔（约 1,100 个 1,536 孔分析板）。该系统配有一个针工具和两个用于添加化合物的声学分配器、四个用于添加试剂的小容量分配器以及三个可实现多种化验检测方法的读板器，可以快速筛选 21 世纪毒理学 (Tox21) 库中的 10,000 种化合物，一式三份。 今年，RSC 团队还完成了 Hamamatsu FDSS 读板机的安装，使 Tox21 程序能够扩展该平台上运行的检测类型，特别是钙通道。 FDSS 本身经过修改，包含 IonField 开发的定制 1536 pintool 清洁器；这是同类产品中首次使用冷等离子体技术代替 DMSO 和其他溶剂进行清洁，由于最大限度地减少了板之间的残留效应，从而提供了更好的数据，并减少了对腐蚀性溶剂的需求，而腐蚀性溶剂在过去会损坏仪器。 NCATS 专家使用与初级筛查系统相同的定制软件来监控和控制该系统。 RNAi 筛选系统有两个用于板运输的机械臂，以及用于存储分析板的培养箱和板堆垛机，以促进系统的连续运行。多通道移液器、低容量分配器、用于洗板的吸气器和两个不同的读板器支持这种以 RNAi 为重点的系统的多种检测方法。 RSC 工作人员还继续支持干细胞翻译实验室 (SCTL)。各个团队成员通过大量设备提供了大量支持，包括自动化组织培养系统、流式细胞仪、细胞分选仪和高内涵成像仪。 特别值得一提的是自动化组织培养平台，该平台已成为所有 NCATS 研究人员的核心能力，现在需求变得如此之大，以至于很可能会订购另一个系统。 这些资源需要大量的专业知识来操作和维护。去年，RSC 已开始充当这些技术的核心设施，使不同的生物学小组能够提高其能力。 与高内涵成像仪相关，由于与前四年的总和相比，能力有所增强，因此去年进行了更多的高内涵测定。 NCATS 校内计划积极参与 NIH 范围内的 COVID-19 救援工作。 RSC 直接参与其中多项关键举措，为多个项目提供项目管理、科学和技术支持。 以下是 RSC 参与的一些 COVID-19 项目的列表： NIH 血清调查：马里兰州贝塞斯达国立卫生研究院已开始进行一项新研究，以确定美国有多少没有确认感染 SARS-CoV-2（引起 2019 年冠状病毒病 (COVID-19) 的病毒）的成年人具有该病毒的抗体。血液中存在抗体表明之前曾感染过。在这次血清调查中，研究人员将收集并分析多达 10,000 名志愿者的血液样本，为流行病学模型提供关键数据。研究结果将有助于阐明新型冠状病毒在美国未被发现的情况下传播的程度，并提供有关哪些社区和人群受影响最严重的见解。 NCATS 和 RSC 在两个方面积极参与这项工作。 对于调查本身，RSC 正在创建一个基于网络的仪表板，该仪表板将直接连接到 NIAID 正在设置的 REDCap 临床试验软件系统。 随着研究结果输入 REDCap，RSC 正在创建一个仪表板，该仪表板将以编程方式提取最新结果并每天更新，以便项目人员监控研究进展并确保达到关键里程碑。 为了分析志愿者提供的样本，RSC 还向 NIBIB 处理结果的实验​​室提供自动化和仪器专业知识来提供协助。 正在运行的诊断是 ELISA 检测，患者的结果在 BMG Labtech PHERAstar 读板机上读出。 这些结果也将更新到相同的基于网络的仪表板，以便项目工作人员也可以看到这些数据。 尽管这个初始阶段从 10,000 名志愿者开始，但需求量很大，而且 RSC 很可能会参与将目前运行 ELISA 检测的手动过程转变为完全自动化的过程。 SARS-CoV-2 CPE 检测（非 GLP）中的化合物测试：该项目涉及发送由 ITRB 化合物管理小组准备的预先点样的检测板，以在南方研究所 (SRI) 进行检测。 该合同已授予（75N95020P00144），五轮检测已完成。 这些数据将通过一个名为 OpenData Portal 的网站公开提供，RSC 将为其提供所有托管支持。 此外，下一轮实验将采用初始实验的结果，并以组合剂量反应方式运行它们，称为矩阵筛选。 所有这些电镀将由 RSC 工作人员进行，并将在我们的 LabCyte Access 系统上进行。 随着更多实验的进行，COVID-19 数据网站上将提供更多数据，并且需要将更多化合物发送到 SRI 和其他可能的 BSL3 设施。