Automated Platform for Fixed Tissue Microdissection

大量存档的人类肿瘤组织，包括癌症活检组织，蕴藏着大量的疾病 相关信息，可能有助于开发新的治疗方法，并有助于我们的基本 对基本生物过程的理解。然而，由于许多原因，资源显然没有得到充分利用。 与检索隐藏在RNA、DNA和DNA中的分子信息相关的技术挑战 锁定在样本中的蛋白质。目前的显微切割平台(特别是单细胞的 解决方案)通常是特定于样本、复杂、昂贵且过度依赖于人工用户干预 在关键步骤。尽管需要，这些限制使得在标准实验室/临床样本处理中进行了整合 工作流程很困难。从福尔马林固定的肿瘤活检组织中快速提取细胞和区域特异性核酸 而存档的组织样本可能是最理想和最具技术挑战性的应用之一。至 为了方便使用存档和临床组织标本，我们建议开发一个自动化平台 (Autopick-MTM)用于高通量固定组织显微解剖。自动从存档的分子检索 临床FFPE组织样本将基于我们最近开发的细胞和组织采集(CTAS) 技术，使用GE的专有方法快速提取组织感兴趣区域(ROI) FFPE组织。重要的是，收集的FFPE ROI被优化用于直接分子询问和 与大多数酶反应兼容，包括用于下一代测序(NGS)的协议。 建议的仪器将具有自动校准、ROI自动识别、采集和分配到 多孔板格式。 我们的初步研究令人信服地证明，样本采集可以通过细胞 分辨率，具有组织分离步骤，确保分离的DNA与一系列下游 方法包括扩增、标记和测序。因为NeuroInDx和Gegr技术都可以 以低成本实施的Autopick-MTM将比现有竞争对手拥有严重的价格优势。 此外，新仪器将保持与新鲜冷冻、活细胞和固定单细胞样本的兼容性， 使Autopick-MTM成为世界上最通用的单细胞采集和组织显微切割平台 市场。这个Fast Track应用程序将测试建议方法的所有关键参数，包括组织 第一阶段的处理和初始仪器架构拟议仪器的商业原型和 将在第二阶段开发和验证完整的工作流程。

Large banks of archived human tumor tissues including cancer biopsies harbor enormous amounts of disease relevant information that may facilitate development of novel therapeutic approaches and assist in our basic understanding of fundamental biological processes. However, the resource is clearly underused, due to a number of technical challenges associated with the retrieval of molecular information hidden in the RNA, DNA and proteins locked inside the samples. Current microdissection platforms (especially those with single cell resolution) are typically sample specific, complex, expensive, and overly dependent on manual user intervention at key steps. Despite the need, these limitations make integration within standard lab/clinical sample processing workflows difficult. Rapid cell- and region-specific nucleic acid extraction from formalin fixed tumor biopsies and archived tissue samples is perhaps one of the most desirable and technically challenging applications. To facilitate the use of the archived and clinical tissue specimens, we propose to develop an automated platform (AutopicK-MTM) for high-throughput fixed tissue microdissection. Automatic molecular retrieval from archived and clinical FFPE tissue samples will be based on our recently developed cell and tissue acquisition (CTAS) technology, using GE’s proprietary approach for rapid extraction of histological regions of interest (ROIs) from FFPE tissues. Importantly, the collected FFPE ROIs are optimized for direct molecular interrogation and compatible with most enzymatic reactions including protocols used for Next Generation Sequencing (NGS). The proposed instrument will feature auto-calibration, ROI auto recognition, collection and dispensing into multiwell plate formats. Our preliminary studies convincingly demonstrate that sample collection may be performed with cellular resolution, with a tissue dissociation step that ensures compatibility of isolated DNA with a range of downstream protocols including amplification, labeling and sequencing. As both NeuroInDx and GEGR technologies can be implemented at low cost, AutopicK-MTM will have a serious price advantage over existing competitors. Moreover, the new instrument will remain compatible with fresh frozen, live cell, and fixed single cell samples, making the AutopicK-MTM the most versatile single cell acquisition and tissue microdissection platform on the market. This Fast Track application will test all critical parameters of the proposed approach including tissue processing and initial instrument architecture in Phase I. Commercial prototype of the proposed instrument and a complete workflow will be developed and validated in Phase II.