3D Microscopy with Ultraviolet Surface Excitation (3D-MUSE)

紫外表面激发 3D 显微镜 (3D-MUSE)

基本信息

批准号：
10620103
负责人：
RICHARD M. LEVENSON
金额：
$ 65.69万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-02 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10620103
关键词：
3-Dimensional Address Anatomy Antibodies Beds Biological Biology Biotechnology Brain Neoplasms California Cell Lineage Cells Clinical Color Computer software Confocal Microscopy Coupled Creativeness Data Set Devices Dimensions Distant Dyes Engineering Face Freezing Functional disorder Histologic Histology Human Image Imaging technology Immobilization Immunohistochemistry Immunotherapy Kidney Label Laboratories Light Magnetic Resonance Imaging Malignant Neoplasms Manuals Measures Methods Microanatomy Microscopy Microtome - medical device Molecular Molecular Analysis Monitor Morphology Mus Nephrology Optics Organ Paraffin Pathology Penetration Perfusion Pharmacologic Substance Phenotype Physiology Prostate Proteins Protocols documentation Reporter Research Research Personnel Resolution Signal Transduction Specimen Stains Standardization Surface System Techniques Technology Testing Three-Dimensional Imaging Time Tissue Embedding Tissues Toxicology Transgenic Animals Universities Visualization absorption cancer immunotherapy clinical application cohort commercialization cost embryo cell empowerment experimental study fluorescence imaging fluorophore gene therapy histological stains image guided image processing imaging agent imaging software imaging system in vivo imaging interest invention laser capture microdissection nanoparticle nanoparticle delivery optical imaging pre-clinical stem cells success theranostics therapeutic nanoparticles tissue preparation ultraviolet vibration virtual

项目摘要

3D Microscopy with Ultraviolet Surface Excitation (3D-MUSE) Summary. We will create powerful techniques for performing 3D histology-quality imaging of small to large volume specimens (e.g., entire mouse or human organ) with extraordinary contrasts to address a very large number of preclinical, biotechnology, and clinical applications. We will combine Microscopy with Ultraviolet Sur- face Excitation (MUSE) (invented at Lawrence Livermore National Laboratory and extended at University of California at Davis), with 3D serial sectioning and block face imaging technologies (developed at Case West- ern Reserve University), to implement 3D-MUSE. Briefly, MUSE fluorescence imaging uses excitation at wave- lengths shorter than 300 nm, a range at which tissue penetration is limited to a few microns deep. This allows one to image—at sub-nuclear resolution—only the very surface of a piece of tissue, making it ideal for serial block-face imaging. Many common histological stains, autofluorescence, and fluorescent proteins excite at these sub-300-nm wavelengths and emit at their familiar wavelengths in the visible range. Because the excita- tion range is distant from emitted signals, images can be captured using a color camera, without additional fil- ters, providing a broad and informative color palette, beyond what is obtained using narrow-band-filter sys- tems. With color-sensitive volume rendering, and combined color and morphology-based segmentation, 3D microanatomy and pathology will be visualized and quantified. Image-guided histology will allow 2D/3D live- time monitoring to guide optional section acquisition for molecular studies using immunohistochemistry and manual or laser-capture micro-dissection. When fully realized, potential applications, out of many, include 3D microanatomy, mouse phenotyping, embryo cell lineage tracking, monitoring therapeutic nanoparticle delivery, as well as studies of cancer physiology, cancer immunotherapy, stem cells, toxicology, and biology. In all of these cases, we anticipate that there will be great value added by the third dimension. We will build on exper- tise in MUSE and block-face imaging systems to create test beds for evaluating 3D-MUSE. We will optimize tissue immobilization, staining, and imaging, and advance MUSE-specific software for 3D segmentation, visu- alization, and analysis. If our research is successful, we will transfer 3D-MUSE technology to commercial enti- ties that can create products for researchers across the world. It will likely be possible to create inexpensive, highly informative 3D-MUSE imaging systems that will empower many applications. Ultimately, the number of researchers using 3D-MUSE around the world will define project success.

紫外表面激发 3D 显微镜 (3D-MUSE) 总结我们将创建强大的技术来执行从小到大的 3D 组织学质量成像。体积标本（例如，整个小鼠或人体器官）具有非凡的对比度，以解决非常大的问题我们将把显微镜与紫外线监视结合起来。面部激发（MUSE）（由劳伦斯利弗莫尔国家实验室发明并在美国大学扩展加利福尼亚州戴维斯），拥有 3D 串行切片和块面成像技术（在 Case West- 开发） ern Reserve University），实施 3D-MUSE 简而言之，MUSE 荧光成像使用波激发。长度短于 300 nm，组织穿透深度仅限于几微米。能够以亚核分辨率仅对一块组织的表面进行成像，使其成为连续成像的理想选择许多常见的组织学染色、自发荧光和荧光蛋白会激发。这些低于 300 纳米的波长并在可见光范围内以其熟悉的波长发射。化范围远离发射信号，可以使用彩色相机捕获图像，无需额外的滤镜 ter，提供了广泛且信息丰富的调色板，超出了使用窄带滤光片系统获得的调色板具有颜色敏感体积渲染以及基于颜色和形态的组合分割、3D 显微解剖学和病理学将被可视化和量化，图像引导的组织学将允许 2D/3D 实时观察。时间监控可指导使用免疫组织化学进行分子研究的可选切片采集手动或激光捕获显微解剖一旦完全实现，潜在的应用包括 3D。显微解剖学、小鼠表型分析、胚胎细胞谱系追踪、监测治疗性纳米颗粒的递送、以及癌症生理学、癌症免疫治疗、干细胞、毒理学和生物学的研究。在这些案例中，我们预计第三维度将带来巨大的附加值，我们将在经验的基础上进一步发展。我们将使用 MUSE 和块面成像系统来创建用于评估 3D-MUSE 的测试平台。组织固定、染色和成像，以及先进的 MUSE 专用软件，用于 3D 分割、可视化如果我们的研究成功，我们将把 3D-MUSE 技术转化为商业实体。可以为世界各地的研究人员创造产品的联系有可能创造出廉价的、信息丰富的 3D-MUSE 成像系统最终将增强许多应用程序的数量。世界各地使用 3D-MUSE 的研究人员将决定项目的成功。