Automated Staining Platform for Immunohistochemistry and In Situ Hybridization

用于免疫组织化学和原位杂交的自动化染色平台

• 批准号: 10177257
• 负责人: Robert Blair
• 金额: $ 22.75万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10177257
• 关键词: Address; Antibodies; Catalogs; Confocal Microscopy; Custom; Data; Detection; Development; Digital Computers; Extramural Activities; Funding; Human; Image Analysis; Immunohistochemistry; In Situ Hybridization; Individual; Intervention; Laboratories; Manuals; Methods; Mus; Primates; Principal Investigator; Process; Protocols documentation; Reagent; Reproducibility; Research; Research Design; Research Personnel; Services; Signal Transduction; Slide; Stains; Techniques; Time; Tissues; United States National Institutes of Health; Vendor; base; digital pathology; experience; improved; nonhuman primate; novel; prevent

PROJECT SUMMARY/ABSTRACT The Ventana Discovery Ultra is an automated stainer for immunohistochemistry (IHC) and in situ hybridization (ISH). The Confocal Microscopy and Image Analysis Core (CMIAC) at the Tulane National Primate Center (TNPRC) has over 30 years of combined experience performing IHC and ISH in nonhuman primate (NHP) tissues. Over the years, IHC and ISH have become fundamental to NIH-funded research at the TNPRC and are widely used by both intramural and extramural investigators. IHC and ISH staining at the TNPRC is currently performed manually by technicians with years of experience. In recent years, the CMIAC has acquired digital pathology and computer-based analyses that allow for the sensitive detection and quantification of IHC and ISH staining. These technological advancements have led to a need for highly reproducible IHC and ISH staining not achievable with the current manual methods. To address this need, nine NIH-funded principal investigators with a total of $19M per year in NIH funding have requested to be major users of the proposed Ventana Discovery Ultra because of its ability to improve the rigor and reproducibility of their IHC and ISH data. The Ventana Discovery Ultra is capable of completing the entire staining process from baking through secondary antibody application, preventing the need for manual intervention at any stage of the process. Multiplex staining protocols that require multiple rounds of primary and secondary antibody staining can also be performed without manual intervention. The Ventana Discovery Ultra is uniquely designed for research, allowing customization of each step of the staining protocol. This is ideal for the development of novel staining protocols, a necessity in NHP research which often requires the optimization of antibodies primarily generated for use in other species (mouse and human). Further, the Ventana Discovery Ultra accepts antibodies, probes, and reagents from third-party vendors, another important feature for the continued use of our extensive catalog of NHP-validated antibodies. In addition, the Ventana Discovery Ultra is one of the few automated stainers capable of performing ISH with signal amplification (RNAScope, ACDbio), a new leading-edge staining technique that is rapidly gaining popularity. This state-of-the-art functionality is necessary to meet the anticipated needs of our users for years to come. The Ventana Discovery Ultra also comes equipped with 30 individual slide bays allowing for the simultaneous staining of 30 unique protocols. This increases efficiency, enhances turnaround time, and facilitates the completion of multiple service requests for a Core laboratory that accommodates 25 investigators on an annual basis.

项目概要/摘要 Ventana Discovery Ultra 是一款用于免疫组织化学 (IHC) 和原位杂交的自动染色机 （ISH）。杜兰国家灵长类动物中心的共焦显微镜和图像分析核心 (CMIAC) (TNPRC) 拥有 30 多年在非人类灵长类动物 (NHP) 中进行 IHC 和 ISH 的综合经验 组织。多年来，IHC 和 ISH 已成为 NIH 资助的 TNPRC 研究的基础，并且 被校内和校外研究者广泛使用。 TNPRC 的 IHC 和 ISH 染色目前正在进行 由具有多年经验的技术人员手动执行。近年来，CMIAC 收购了数字 病理学和基于计算机的分析，可对 IHC 和 ISH 进行灵敏检测和定量 染色。这些技术进步导致需要高度可重复的 IHC 和 ISH 染色，而不是 通过当前的手动方法可以实现。为了满足这一需求，九名 NIH 资助的主要研究人员 NIH 每年请求总计 1900 万美元的资金成为拟议的 Ventana Discovery 的主要用户 Ultra 是因为它能够提高 IHC 和 ISH 数据的严谨性和可重复性。文塔纳 Discovery Ultra能够完成从烘焙到二抗的整个染色过程 应用程序，无需在流程的任何阶段进行手动干预。多重染色方案 需要多轮一抗和二抗染色的操作也可以在无需手动操作的情况下进行 干涉。 Ventana Discovery Ultra 专为研究而设计，允许定制每个步骤 染色方案。这非常适合开发新型染色方案，这是 NHP 研究的必需品 这通常需要优化主要用于其他物种（小鼠和 人类）。此外，Ventana Discovery Ultra 接受来自第三方供应商的抗体、探针和试剂， 继续使用我们广泛的 NHP 验证抗体目录的另一个重要特征。此外， Ventana Discovery Ultra 是少数能够执行 ISH 信号的自动染色机之一 扩增（RNAScope、ACDbio），一种新的前沿染色技术，正在迅速普及。这 为了满足用户未来几年的预期需求，最先进的功能是必要的。这 Ventana Discovery Ultra 还配备了 30 个独立的载玻片托架，可同时进行染色 30 个独特的协议。这提高了效率，缩短了周转时间，并有利于完成 针对每年容纳 25 名研究人员的核心实验室提出多项服务请求。