quantitative microscopy-based rapid phenotyping and screening

基于定量显微镜的快速表型分析和筛选

• 批准号: 8638024
• 负责人: Hang Lu
• 金额: $ 27.42万
• 依托单位: GEORGIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8638024
• 关键词: Age; Aging; Algorithm Design; Algorithms; Animals; Apoptosis; Area; Automation; Biological Models; Caenorhabditis elegans; Classification; Collection; Computer Assisted; Computers; Data; Development; Developmental Cell Biology; Device Designs; Devices; Disease; Drosophila genus; Embryo; Engineering; Enhancers; Ensure; Environment; Event; Eye; Fill-It; Fluorescence; Genes; Genetic; Genetic Models; Genetic Research; Genetic Screening; Goals; Image; Image Analysis; Imaging Techniques; Imaging technology; Larva; Lead; Length; Life; Link; Maintenance; Manuals; Measures; Mental disorders; Methods; Metric; Microfluidic Microchips; Microfluidics; Microscopy; Modeling; Molecular; Molecular Biology; Motor Neurons; Mutagenesis; Nematoda; Nerve Degeneration; Nervous system structure; Neurobiology; Neurodegenerative Disorders; Neurons; Neurosciences; Nobel Prize; Optics; Organism; Pathway interactions; Pattern; Phenotype; Phylogeny; Positioning Attribute; Process; Quantitative Microscopy; RNA Interference; Regulatory Pathway; Research; Shapes; Sorting - Cell Movement; Speed; Standardization; Structure; Synapses; Syncope; System; Techniques; Technology; Testing; Therapeutic; Time; Training; Visual; Work; Zebrafish; age related; base; computerized; design; fly; forward genetics; genome sequencing; human disease; image processing; improved; in vivo; in vivo imaging; innovation; interest; mutant; nerve injury; nervous system disorder; neurogenetics; new technology; novel; programs; public health relevance; research study; screening; synaptogenesis; therapeutic target; tool; trait

DESCRIPTION (provided by applicant): Synapses are most fundamental to the function of a nervous system. C. elegans is an excellent genetic model system for finding genes and elucidating pathways because of its sequenced genome and the abundance of molecular biology tools and mutants. Due to the simplicity of its nervous system, many breakthroughs have been made in C. elegans for understanding molecular mechanisms in the patterning of the nervous system and synapse development. The current bottlenecks, however, are in the manual and non-quantitative techniques such as visual screens, often limiting both the throughput of the experiments and the phenotypes one can examine. Our long-term objective is to develop technologies to understand how genes, age, and the environment together define and continue to remodel the nervous system of an organism. Microtechnologies are ideal for studies of C. elegans neuroscience because of the relevant length scales and the possibility for automation; similarly quantitative imaging techniques are key to deciphering molecular mechanisms. The objective of this R01 project is to engineer micro devices for large-scale live imaging and quantitative imaging technologies in order to study synapse development in an in vivo system. Genes and pathways emerging from this study could potentially become targets of therapeutics in neurological disorders. We hypothesize that quantitative microscopy-based approaches can indeed enable identification of novel genes and pathways that conventional approaches cannot. The first component of this project is to develop on-chip rapid and high-content in vivo imaging techniques, and in parallel to develop algorithms and quantitative measures for the analysis of such high-content data. The second component of the project is to perform screens and studies using these novel technologies. The approach is innovative because the technology developed here dramatically increases the capabilities of existing imaging and screening tools by several orders of magnitude in speed and much more sensitive and accurate than conventional manual approaches. The proposed research is significant because it fills the urgent need in high-throughput and high-content screens as well as identifying novel genes and pathways. In addition, besides the contribution to the specific neurobiology, the technologies are widely applicable to areas such as developmental cell biology, and to other small organisms such as fly larvae and zebrafish embryos.

描述（由申请人提供）：突触是神经系统功能最重要的。 秀丽隐杆线虫是一个出色的遗传模型系统，用于查找基因和阐明途径，因为其测序的基因组以及丰富的分子生物学工具和突变体。 由于其神经系统的简单性，在秀丽隐杆线虫中已经取得了许多突破，以了解神经系统和突触发展的分子机制。 但是，当前的瓶颈是在手动和非量化技术（例如视觉屏幕）中，通常限制了实验的吞吐量和可以检查的表型。 我们的长期目标是开发技术，以了解基因，年龄和环境如何共同定义并继续重塑生物体的神经系统。 由于相关的长度尺度和自动化的可能性，微技术是对秀丽隐杆线虫神经科学研究的理想选择。类似地，定量成像技术是破译分子机制的关键。 该R01项目的目的是设计用于大规模实时成像和定量成像技术的微设备，以研究体内系统中的突触发展。 这项研究中出现的基因和途径可能会成为神经系统疾病中治疗剂的靶标。 我们假设基于定量显微镜的方法确实可以鉴定常规方法无法鉴定新的基因和途径。 该项目的第一个组成部分是在体内成像技术中开发芯片快速和高含量，并并行开发算法和定量测量，以分析此类高含量数据。 该项目的第二个组成部分是使用这些新技术进行筛选和研究。 这种方法具有创新性，因为这里开发的技术大大提高了现有成像和筛选工具的功能，而不是传统的手动方法的速度几个数量级，并且更敏感和准确。 拟议的研究很重要，因为它填补了高通量和高含量屏幕的迫切需求，并确定了新颖的基因和途径。 此外，除了对特定神经生物学的贡献外，这些技术还广泛适用于发育细胞生物学等领域，以及其他小生物（例如蝇幼虫和斑马鱼胚胎）。