NeuroExM

神经ExM

• 批准号: 10686269
• 负责人: JACOB R GLASER
• 金额: $ 99.56万
• 依托单位: MICROBRIGHTFIELD, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-02 至 2025-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10686269
• 关键词: 3-Dimensional; Aging; Antibodies; Axon; Benchmarking; Biological; Biological Preservation; Biological Sciences; Biotechnology; Brain; Brain Diseases; Cell physiology; Central Nervous System; Classification; Collaborations; Communities; Complex; Complex Analysis; Computer software; Computers; Data; Dendrites; Dendritic Spines; Detection; Development; Disease; Drug Addiction; Feasibility Studies; Image; In Situ; Individual; Investigation; Knowledge; Label; Light; Manuals; Maps; Messenger RNA; Microscope; Microscopy; Modeling; Molecular; Morphology; Neurodegenerative Disorders; Neurodevelopmental Disorder; Neurons; Neurosciences Research; Performance; Peripheral; Phase; Population; Population Analysis; Population Distributions; Preparation; Process; Production; Proteins; Proteome; Research Personnel; Resolution; Running; Sampling; Scientist; Societies; Spatial Distribution; Specimen; Stains; Structure; Subcellular structure; Synapses; System; Techniques; Technology; Testing; Three-Dimensional Image; Time; Tissue Expansion; Tissues; Translations; Validation; Visualization; Work; age effect; complex biological systems; design; drug development; drug discovery; fighting; fluorophore; high dimensionality; image registration; improved; innovation; interest; microscopic imaging; nanoscale; nervous system disorder; neuronal cell body; neuropsychiatric disorder; new technology; next generation; novel; pharmacologic; prevent; prototype; reconstruction; research and development; research study; software development; terabyte; tissue processing; transcriptome; translational neuroscience; treatment strategy; usability

Abstract This project describes the development of NeuroExM™, a highly innovative system for performing comprehensive spatial distribution analysis of populations of messenger RNAs (mRNAs) and proteins in tissue processed for expansion microscopy (ExM)). The groundbreaking technological advantage of ExM, which was recently developed by Dr. Edward S. Boyden (Dept. Biol. Engin., Media Lab and Dept. Brain Cognit. Sci., MIT, Cambridge, MA) and colleagues, is the ability to isotropically expand tissue and increase the size of the biological structures. This allows nanoscale-resolution, light-microscopic imaging of small objects that are too small to be resolved without expansion due to the diffraction limit of light. Among other benefits, ExM allows those small structures to be imaged with a wider range of microscopy techniques. Processing tissue for ExM also allows repeated hybridization (for investigations of mRNAs) and/or repeated antibody staining (for investigations of proteins) of the same tissue, combined with repeated microscopic imaging rounds. Each round yields adjacent, high-magnification, single field-of-view image stacks, consisting of at least one morphology reference channel showing neuronal sub-cellular structures (somas, axons, dendrites, dendritic spines, synapses) as well as one or several info channels showing mRNAs and/or proteins. Comprehensive analysis of the spatial distribution of populations of mRNAs and proteins in neurons in situ requires assembling the image stacks of all performed rounds into a single, seamless and aligned, three-dimensional (3D) ExM image, which is high-dimensional and can be several terabytes in size. However, this presents a number of computational challenges with respect to microscopy image registration, segmentation and analysis. The game-changing innovation in NeuroExM is the ability to perform all of these tasks without the need to have a computer scientist on staff to run the existing, individual lab-based software scripts developed for each step of this kind of complex analysis. This is made possible by implementing a number of significant technical innovations into NeuroExM. Based on pilot work performed in collaboration with the Boyden lab during preparation of this proposal, we are convinced that NeuroExM will make a significant impact on the field of neuroscience research. Specifically, the combination of ExM and NeuroExM will enable substantial advancements in research studies focusing on alterations in the spatial transcriptome and proteome of neurons associated with neurodevelopmental, neuropsychiatric, neurodegenerative and neurological disorders as well as in aging research and drug development. Ultimately, this will result in an improved basis for developing novel treatment strategies for a wide spectrum of complex brain diseases. In Phase I we will demonstrate feasibility of this novel technology by developing prototype software; work in Phase II will focus on creating the full functionality of NeuroExM for commercial release. We will perform extensive feasibility studies, product validation and usability studies of NeuroExM in close collaboration with the Boyden lab. A competing technology is not available.

抽象的 该项目描述了 NeuroExM™ 的开发，这是一个高度创新的系统，用于执行 组织中信使 RNA (mRNA) 和蛋白质群体的综合空间分布分析 用于扩展显微镜（ExM））。 ExM 的突破性技术优势是 最近由 Edward S. Boyden 博士（麻省理工学院生物工程系、媒体实验室和脑认知科学系）开发， 马萨诸塞州剑桥市）及其同事的研究成果是，能够各向同性地扩展组织并增加生物体的尺寸。 结构。这使得能够对太小而无法观测的小物体进行纳米级分辨率的光学显微成像。 由于光的衍射极限，在没有膨胀的情况下解析。除其他好处外，ExM 还允许那些小型企业 使用更广泛的显微镜技术对结构进行成像。处理 ExM 组织还允许 重复杂交（用于研究 mRNA）和/或重复抗体染色（用于研究 蛋白质），结合重复的显微成像轮次。每轮产生相邻的， 高放大率、单视场图像堆栈，由至少一个形态参考通道组成 显示神经元亚细胞结构（体细胞、轴突、树突、树突棘、突触）以及一种 或几个显示 mRNA 和/或蛋白质的信息通道。空间分布综合分析 原位神经元中的 mRNA 和蛋白质群体需要组装所有执行的图像堆栈 舍入为单个、无缝且对齐的三维 (3D) ExM 图像，该图像是高维且 大小可能有几 TB。然而，这提出了许多计算挑战 显微镜图像配准、分割和分析。 NeuroExM 改变游戏规则的创新是 能够执行所有这些任务，而无需配备计算机科学家来运行现有的、 为这种复杂分析的每个步骤开发的单独的基于实验室的软件脚本。这是做的 通过在 NeuroExM 中实施许多重大技术创新，这成为可能。基于试点工作 在准备本提案期间与博伊登实验室合作进行，我们确信 NeuroExM 将对神经科学研究领域产生重大影响。具体来说，组合 ExM 和 NeuroExM 将使专注于改变的研究取得实质性进展 与神经发育、神经精神、 神经退行性和神经系统疾病以及衰老研究和药物开发。最终， 这将为开发针对广泛的复杂疾病的新治疗策略奠定更好的基础 脑部疾病。在第一阶段，我们将通过开发原型来证明这项新技术的可行性 软件;第二阶段的工作将侧重于创建 NeuroExM 的全部功能以供商业发布。我们 将密切对 NeuroExM 进行广泛的可行性研究、产品验证和可用性研究 与博伊登实验室的合作。没有可用的竞争技术。