Instrumentation platform for 3D pathology with open-top light-sheet microscopy

具有开顶光片显微镜的 3D 病理学仪器平台

• 批准号: 10178401
• 负责人: Jonathan T.C. Liu
• 金额: $ 55.87万
• 依托单位: UNIVERSITY OF WASHINGTON
• 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 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10178401
• 关键词: 3-Dimensional; Adoption; Anatomy; Antibodies; Architecture; Benchmarking; Biological Assay; Biological Markers; Biopsy; Biopsy Specimen; Cells; Clinical; Clinical Research; Costs and Benefits; DNA; Data; Development; Devices; Diagnosis; Diagnostic; Disease; Dissection; Excision biopsy; Fluorescent Dyes; Formalin; Funding Agency; Future; Generations; Genomics; Glass; Gold; Healthcare; Histology; Histopathology; Hybrids; Image; Imaging technology; Immersion; Institution; Kidney Diseases; Label; Light; Malignant - descriptor; Malignant Neoplasms; Methods; Microfluidics; Microscope; Microscopy; Motivation; Neurodegenerative Disorders; Nucleic Acids; Operative Surgical Procedures; Optics; Paraffin Embedding; Pathologic; Pathologist; Pathology; Pathway interactions; Patients; Performance; Precision therapeutics; Prediction of Response to Therapy; Preparation; Process; Protocols documentation; Public Health; Publishing; Refractive Indices; Reproducibility; Research Personnel; Resolution; Sampling; Services; Slide; Specimen; Speed; Stains; Standardization; Structure; System; Techniques; Technology; Thick; Three-Dimensional Image; Tissue Sample; Tissues; analog; base; clinical decision support; deep sequencing; design; human tissue; image guided; imaging modality; improved; indexing; innovation; instrumentation; lightspeed; microscopic imaging; molecular diagnostics; novel; personalized medicine; prognostic; prototype; sequencing platform; tissue preparation; tool; translational impact

Summary The clinical gold-standard method for interrogating tissue specimens, slide-based (2D) histopathology, is based on centuries-old technologies with many inherent limitations. Recent technological advances have demonstrated the feasibility of achieving high-throughput slide-free 3D histology of biopsy and surgical specimens. In comparison to conventional slide-based histology, nondestructive 3D histology has the potential to provide a transformative improvement in diagnostic pathology performance for a number of reasons: (1) vastly greater (>100X) sampling of tissue specimens, (2) volumetric imaging of 3D cell distributions and tissue structures that are prognostic and predictive, (3) nondestructive imaging, which allows valuable biopsy specimens to be used for downstream biomarker assessment, and (4) a simplified process with cost benefits for healthcare institutions and payers. In recent years, we have developed a technology, open-top light-sheet (OTLS) microscopy, to enable high-throughput nondestructive 3D histology of ex vivo specimens. Our first generations of OTLS microscopes and imaging protocols demonstrated the ability to reliably image a variety of optically cleared clinical tissue specimens (surgical excisions and biopsies) in a nondestructive manner that does not interfere with conventional pathology methods. Here, we propose to develop a multi-resolution hybrid OTLS microscope (Aim 1), based on a novel non-orthogonal dual-objective (NODO) architecture, which will be superior in every regard to our previous systems, including resolution (and range of resolutions), imaging depth, and compatibility with nearly all clearing/labeling protocols and sample-holder materials (insensitivity to refractive-index mismatch). Furthermore, we will develop innovative pre-imaging methods to automate and standardize the tissue-labeling and clearing process for a robust fluorescent analog of H&E staining (Aim 2). Finally, we will develop post- imaging technologies for image-guided macro-dissection of thick tissues, which we will show has the ability to significantly improve the sensitivity of genomic assays (Aim 3). Collectively, our project aims are designed to extend current 2D pathology workflows into 3D to minimize clinical-adoption barriers. A rapid translational pathway exists through a 3D-pathology-services company (Lightspeed Microscopy Inc.) that has licensed our entire 3D pathology IP portfolio. As part of this larger translational effort, clinical studies are ongoing in our labs, along with development of AI-analysis methods for clinical decision-support (i.e. prognostication and prediction of treatment response). The instrumentation platform developed in this project will directly support a number of future disease-focused clinical studies to demonstrate the value of 3D pathology for the precision treatment of diverse conditions such as kidney disease, neurodegenerative diseases, and various forms of cancer.

概括 用于检查组织样本的临床金标准方法，即基于载玻片的 (2D) 组织病理学，基于 基于具有许多固有局限性的数百年历史的技术。最近的技术进步已经证明 实现活检和手术标本高通量无载玻片 3D 组织学的可行性。在 与传统的基于载玻片的组织学相比，非破坏性 3D 组织学有可能提供 诊断病理学性能的革命性改进有多种原因：(1) 大大提高 (>100X) 组织样本采样，(2) 3D 细胞分布和组织结构的体积成像 具有预后和预测作用，(3) 无损成像，可以使用有价值的活检标本 用于下游生物标志物评估，以及 (4) 简化流程，为医疗机构带来成本效益 和付款人。近年来，我们开发了一种技术，即开顶光片 (OTLS) 显微镜， 实现离体标本的高通量无损 3D 组织学分析。我们的第一代 OTLS 显微镜和成像协议证明了能够可靠地对各种光学透明的临床图像进行成像 以不干扰的非破坏性方式获取组织标本（手术切除和活检） 常规病理学方法。在这里，我们建议开发一种多分辨率混合 OTLS 显微镜（Aim 1）、基于新颖的非正交双目标（NODO）架构，该架构在各方面都将更加优越 与我们之前的系统相比，包括分辨率（和分辨率范围）、成像深度以及与 几乎所有的透明/标签协议和样品支架材料（对折射率不匹配不敏感）。 此外，我们将开发创新的预成像方法，以实现组织标记的自动化和标准化 H&E 染色的强大荧光类似物的清除过程（目标 2）。最后，我们将开发后期 用于图像引导厚组织宏观解剖的成像技术，我们将展示该技术能够 显着提高基因组检测的灵敏度（目标 3）。总的来说，我们的项目目标旨在 将当前的 2D 病理学工作流程扩展到 3D，以最大限度地减少临床采用障碍。快速翻译 途径存在于一家 3D 病理学服务公司 (Lightspeed Microscopy Inc.)，该公司已获得我们的许可 整个 3D 病理学 IP 产品组合。作为这项更大的转化工作的一部分，我们的实验室正在进行临床研究， 随着用于临床决策支持的人工智能分析方法的发展（即预测和预测） 治疗反应）。本项目开发的仪器平台将直接支持多种 未来以疾病为中心的临床研究将证明 3D 病理学对于精准治疗的价值 多种疾病，如肾脏疾病、神经退行性疾病和各种癌症。