Automated Staining Platform for Immunohistochemistry and In Situ Hybridization

项目摘要/摘要 Ventana Discovery Ultra是一种用于免疫组织化学(IHC)和原位杂交的自动染色设备 (差不多)。杜兰国家灵长类中心的共焦显微镜和图像分析核心(CMIAC) (TNPRC)有30多年在非人灵长类动物(NHP)上进行IHC和ISH的综合经验 纸巾。多年来，IHC和ISH已经成为国家卫生研究院资助的TNPRC研究的基础，并正在 被校内和校外调查人员广泛使用。TNPRC的IHC和ISH染色目前 由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，允许灵敏地检测和量化IHC和ISH 染色。这些技术的进步导致了对高度重复性的IHC和ISH染色的需求 目前的手工方法是可以实现的。为了满足这一需求，由美国国立卫生研究院资助的九名首席调查人员 美国国立卫生研究院每年总共提供1900万美元的资金，要求成为拟议的文塔纳发现号的主要用户 ULTRA是因为它有能力提高其IHC和ISH数据的严密性和重复性。文塔纳 Discovery Ultra能够完成从烘焙到二次抗体的整个染色过程 应用程序，无需在过程的任何阶段进行人工干预。多重染色方案 需要多轮一次和二次抗体染色也可以不需要人工进行 干预。Ventana Discovery Ultra专为研究而设计，允许定制每个步骤 染色方案。这对于开发新的染色方案是理想的，这是NHP研究中的必备条件 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还接受第三方供应商提供的抗体、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是为数不多的能够使用SIGNAL执行ISH的自动染色设备之一 扩增(RNAScope，ACDbio)，这是一种正在迅速普及的新的前沿染色技术。这 最先进的功能是必要的，以满足我们用户在未来几年的预期需求。这个 Ventana Discovery Ultra还配备了30个单独的幻灯片托架，可以同时进行染色 30种独特的协议。这提高了效率，缩短了周转时间，并有助于完成 对每年可容纳25名调查人员的核心实验室提出多项服务请求。

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.