Automated Platform for Fixed Tissue Microdissection

固定组织显微切割自动化平台

• 批准号: 9488425
• 负责人: Stanislav Karsten
• 金额: $ 71.84万
• 依托单位: NEUROINDX INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-21 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9488425
• 关键词: Admixture; Alleles; Architecture; Archives; Attention; Automation; Biochemical Reaction; Biological Assay; Biological Process; Biopsy; Buffers; Calibration; Cells; Clinical; Clinical Services; Collection; Complex; Computer software; Consult; Consultations; DNA; Deposition; Development; Diagnostic; Digestion; Disease; Dissection; Dissociation; Ensure; Evaluation; Feedback; Formalin; Freezing; Goals; Histologic; Human; Injections; Intervention; Label; Lasers; Legal patent; Licensing; Malignant Neoplasms; Manuals; Methods; Microdissection; Molecular; Molecular Analysis; Mutation Detection; Myeloma Proteins; Normal tissue morphology; Nucleic Acids; Paraffin Embedding; Pharmaceutical Services; Phase; Preparation; Price; Procedures; Process; Proteins; Protocols documentation; RNA; Reagent; Reproducibility; Research; Resolution; Resources; Retrieval; Running; Sampling; Scheme; Series; Slide; Small Business Innovation Research Grant; Source; Specimen; Speed; Stains; System; Technology; Testing; Time; Tissue Embedding; Tissue Microarray; Tissue Sample; Tissue imaging; Tissues; Tube; Tumor Tissue; Validation; Visualization software; base; clinical application; clinically relevant; commercialization; cost; cost effective; digital imaging; digital pathology; drug development; instrument; interest; manufacturing process; melanoma; mutant; next generation sequencing; novel therapeutic intervention; oncology; prototype; sample collection; single cell analysis; software development; statistics; tissue culture; tissue processing; tumor

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.

大量存档的人类肿瘤组织，包括癌症活检，有大量疾病 相关信息可能有助于开发新的治疗方法并帮助我们的基本 了解基本生物学过程。但是，由于一个数字，资源显然没有用 与RNA，DNA和 蛋白质锁定在样品中。当前的微分解平台（尤其是具有单个单元格的平台 分辨率）通常是特定，复杂，昂贵且过于依赖手动用户干预 在关键步骤中。尽管需要，这些限制仍会在标准实验室/临床样本处理中集成 工作流程困难。从福尔马林固定肿瘤活检中提取快速细胞和区域特异性核酸 存档的组织样品可能是最令人期望，最具技术挑战的应用之一。到 促进存档和临床组织标本的使用，我们建议开发自动化平台 （Autopick-MTM）用于高通量固定组织显微解剖。自动分子从存档中检索 临床FFPE组织样品将基于我们最近开发的细胞和组织采集（CTA） 技术，使用GE的专有方法来快速提取感兴趣的组织学区域（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 与大多数酶促反应兼容，包括用于下一代测序（NGS）的方案。 拟议的仪器将具有自动校准，ROI自动识别，收集和分配到 多韦板格式。 我们的初步研究令人信服地表明，可以用细胞进行样品收集 分辨率，具有组织解离步骤，可确保孤立的DNA与一系列下游的兼容性 协议，包括放大，标记和测序。由于神经因素和GEGR技术都可以 自动驾驶MTM以低成本实施将比现有竞争对手具有严重的价格优势。 此外，新仪器将与新鲜的冷冻，活细胞和固定单细胞样品保持兼容， 使AutoPick-MTM成为最通用的单细胞采集和组织微解剖平台 市场。此快速应用将测试所提出方法的所有关键参数，包括组织 第1期的处理和初始仪器架构 在第二阶段将开发和验证完整的工作流程。