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 基于幻灯片（2D）组织病理学审查组织标本的临床金标准方法是基于 在具有许多固有局限性的数百年技术中。最近的技术进步已经证明 实现活检和手术标本的高通量幻灯片3D组织学的可行性。在 与传统的基于幻灯片的组织学比较，非破坏性3D组织学具有提供的潜力 由于多种原因 （> 100倍）组织样品的采样，（2）3D细胞分布和组织结构的体积成像 具有预后和预测性，（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）简化的过程，为医疗机构带来成本收益 和付款人。近年来，我们开发了一项技术，开放式灯页（OTL）显微镜，以便 启用体内标本的高通量非破坏性3D组织学。我们的第一代OTL 显微镜和成像方案证明了能够可靠地图像各种光学清除的临床的能力 组织标本（手术切除和活检）以不受干扰的方式不干扰 常规病理方法。在这里，我们建议开发多分辨率混合OTLS显微镜（AIM 1），基于一种新颖的非正交双目标（NODO）架构，在各个方面都将是优越的 到我们以前的系统，包括分辨率（和分辨率范围），成像深度以及与 几乎所有清除/标记方案和样品持有材料（对折射率不匹配的不敏感）。 此外，我们将开发创新的预知方法来自动化和标准化组织标签 以及清除H＆E染色的强大荧光类似物（AIM 2）的清除过程。最后，我们将发展后 图像引导的厚组织宏观划痕的成像技术，我们将表明具有能力 显着提高基因组测定的敏感性（AIM 3）。总的来说，我们的项目旨在 将当前的2D病理工作流扩展到3D，以最大程度地减少临床障碍。快速翻译 途径通过3D PARCHOLOGY-SERVICES COMPANY（LIGHTSPEED MICROSCOPY INC.），该公司已获得许可 整个3D病理IP投资组合。作为这项更大的翻译工作的一部分，我们的实验室正在进行临床研究， 以及开发用于临床决策支持的AI-Analysis方法（即预测和预测 治疗反应）。该项目开发的仪器平台将直接支持许多 未来以疾病为中心的临床研究证明了3D病理学的价值 肾脏疾病，神经退行性疾病和各种形式的癌症等多种疾病。