Automated Staining Platform for Immunohistochemistry and In Situ Hybridization

项目总结/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仪 （ISH）。杜兰国家灵长类中心的共聚焦显微镜和图像分析核心（CMIAC） （TNPRC）在非人灵长类动物（NHP）中进行IHC和ISH的综合经验超过30年 组织中多年来，IHC和ISH已成为NIH资助的研究在TNPRC的基础， 被校内和校外的研究人员广泛使用。目前，TNPRC的IHC和ISH染色 由具有多年经验的技术人员手动执行。近年来，CMIAC已经收购了数字 病理学和基于计算机的分析，允许灵敏地检测和定量IHC和ISH 染色这些技术进步导致需要高度可重复的IHC和ISH染色， 这是目前手工操作的方法。为了满足这一需求，9名NIH资助的主要研究人员， NIH每年总计1900万美元的资金要求成为拟议的Ventana Discovery的主要用户 Ultra因为它能够提高IHC和ISH数据的精确性和重现性。温塔娜潜艇 Discovery Ultra能够完成从烘焙到二抗的整个染色过程 应用程序，防止在该过程的任何阶段需要手动干预。多重染色方案 需要多轮第一和第二抗体染色也可以在没有人工 干预Ventana Discovery Ultra专为研究而设计，允许定制每个步骤 染色方案的一部分。这是理想的新染色方案的发展，在NHP研究的必要性 这通常需要优化主要用于其它物种（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， 这是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，本发明还提供了一种方法， Ventana Discovery Ultra是为数不多的能够用信号进行ISH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 最先进的功能是必要的，以满足我们的用户在未来几年的预期需求。的 Ventana Discovery Ultra还配备了30个单独的载玻片托架，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，并促进了 每年可容纳25名研究者的核心实验室的多次服务请求。

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.