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，这是一种高度创新的系统 组织中的Messenger RNA（mRNA）和组织中蛋白质种群的全面空间分布分析 处理用于扩展显微镜（EXM））。 EXM的开创性技术优势， 最近由Edward S. Boyden博士开发（Biol。Engin。 剑桥，马萨诸塞州和同事是各向同性扩展组织并增加生物学大小的能力 结构。这允许纳米级分辨率，小物体的轻微微观成像，这些成像太小而无法为 由于光的衍射极限而无需扩展而解决。除其他好处外，EXM允许那些小的 用更广泛的显微镜技术成像的结构。 EXM的处理组织也允许 重复杂交（用于mRNA的投资）和/或重复的抗体染色（用于投资 同一组织的蛋白质），与重复的微观成像弹相结合。每个回合屈服相邻， 高磁化，单个视野图像堆栈，由至少一个形态参考通道组成 显示神经元亚细胞结构（躯体，轴突，树突，树突状刺，突触）以及一个 或几个信息渠道显示mRNA和/或蛋白质。对空间分布的全面分析 神经元中mRNA和蛋白质的种群需要组装所有执行的图像堆栈 圆形分为一个，无缝且对齐的三维（3D）EXM图像，该图像是高维的 可以是几个大小的trabytes。但是，这对 显微镜图像注册，分割和分析。 NeuroExm中改变游戏规则的创新是 能够执行所有这些任务的能力，而无需让计算机科学家在工作人员中运行现有的， 基于实验室的单个软件脚本为这种复杂分析的每个步骤开发。这是做出的 通过将许多重要的技术创新实施到神经外观中。基于飞行员工作 在准备此提案期间，与博伊登实验室合作进行，我们坚信 NeuroExm将对神经科学研究领域产生重大影响。具体而言，结合 EXM和NeuroExm将在研究研究的研究中实现重大进步，重点是 神经发育，神经精神病学相关的神经元的空间转录组和蛋白质组， 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， 这将为开发各种复杂范围的新型治疗策略提供改进的基础 脑部疾病。在第一阶段，我们将通过开发原型来证明这种新技术的可行性 软件;第二阶段的工作将着重于创建神经Exm的全部功能，以供商业发布。我们 将对神经EXM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。竞争技术不可用。