Real-time imaging of dynamic biological processes at nanometer spatial resolution

以纳米空间分辨率对动态生物过程进行实时成像

• 批准号: 7826759
• 负责人: James E Evans
• 金额: $ 50万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7826759
• 关键词: Address; Area; Biological; Biological Process; Cell physiology; Cells; Cellular biology; Characteristics; Chemicals; Couples; Dependence; Development; Dynamin; Electron Beam; Electron Microscope; Electrons; Environment; Equilibrium; Event; Filament; Goals; Guanosine Triphosphate Phosphohydrolases; Image; Imaging Techniques; In Situ; Individual; Kinesin; Label; Lasers; Life; Liquid substance; Macromolecular Complexes; Mediating; Membrane; Methods; Microtubule-Associated Proteins; Microtubules; Mitochondria; Mitosis; Molecular; Molecular Structure; Motor; Movement; Nucleotides; Organelles; Paclitaxel; Physiologic pulse; Play; Process; Proteins; Radiation; Radiation Tolerance; Relative (related person); Research; Resolution; Role; Sampling; Specimen; Staging; Structure; System; Techniques; Technology; Testing; Time; Work; cancer type; chemotherapeutic agent; electron crystallography; improved; instrument; millisecond; movie; nanometer; nanosecond; novel strategies; prevent; public health relevance; reconstruction; research study; spatiotemporal; three dimensional structure; transmission process

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area (06) Enabling Technologies and specific Challenge Topic, 06-GM-101: Structural analysis of macromolecular complexes. The focus of this proposal is the development and optimization of a novel approach for high-resolution direct imaging of biological processes as they occur in real-time. The "live" movies will be acquired using a Dynamic Transmission Electron Microscope that couples the spatial resolution characteristics of an electron microscope with the temporal resolution of ultrafast Lasers. This first-of-its-kind instrument for imaging biological samples will be capable of sub-nanometer and nanosecond spatiotemporal resolution and will increase the combined attainable resolution five to eight orders of magnitude over current techniques. We further postulate that the resolution limiting effects observed during conventional cryo-EM imaging of biological samples arise from long-term specimen and electron beam interactions, and will be alleviated by using the Dynamic TEM with ultrafast imaging pulses. Mitigating these effects will result in improved three dimensional structure determination of macromolecular complexes. The impact of this research has the potential to elucidate new levels of understanding regarding microtubule and mitochondrial dynamics and the mechanisms involved with both normal and abnormal cellular processes. PUBLIC HEALTH RELEVANCE: Our work will provide the first high-resolution direct imaging of biological processes as they occur in real-time. These "live" movies will be acquired using a Dynamic Transmission Electron Microscope that combines characteristics of an atomic resolution electron microscope with ultrafast Lasers to study the structure of macromolecular complexes. This first-of-its-kind instrument will increase the spatial and temporal resolution for observing dynamic biological processes by five to eight orders of magnitude over current techniques.

描述（由申请人提供）：本申请涉及广泛的挑战领域 (06) 实现技术和具体挑战主题，06-GM-101：大分子复合物的结构分析。该提案的重点是开发和优化一种新方法，用于实时发生的生物过程的高分辨率直接成像。 “现场”电影将使用动态透射电子显微镜采集，该显微镜将电子显微镜的空间分辨率特性与超快激光器的时间分辨率相结合。这种用于生物样本成像的同类仪器将能够实现亚纳米和纳秒时空分辨率，并将可达到的综合分辨率比现有技术提高五到八个数量级。我们进一步假设，在生物样品的传统冷冻电镜成像过程中观察到的分辨率限制效应是由长期样本和电子束相互作用引起的，并且通过使用具有超快成像脉冲的动态 TEM 可以减轻这种影响。减轻这些影响将改善大分子复合物的三维结构测定。这项研究的影响有可能阐明对微管和线粒体动力学以及涉及正常和异常细胞过程的机制的新水平的理解。 公共健康相关性：我们的工作将提供第一个实时发生的生物过程的高分辨率直接成像。这些“现场”电影将使用动态透射电子显微镜采集，该显微镜将原子分辨率电子显微镜的特性与超快激光器相结合，以研究大分子复合物的结构。这种首创的仪器将把观察动态生物过程的空间和时间分辨率比现有技术提高五到八个数量级。