Ceramic Electron Microscopy Grids for Cell Culturing and Multiscale Imaging

用于细胞培养和多尺度成像的陶瓷电子显微镜网格

• 批准号: 7909857
• 负责人: Brent Joseph Lutz
• 金额: $ 10万
• 依托单位: SYNKERA TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2010-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7909857
• 关键词: Address; Adhesions; Architecture; Arts; Biological; Biophysics; Carbon; Cell Culture Techniques; Cells; Cellular biology; Ceramics; Colorado; Cryoelectron Microscopy; Cultured Cells; Data; Data Quality; Development; Dimensions; Electron Microscopy; Evaluation; Film; Foundations; Freezing; Generations; Goals; Gold; Government; Image; Imaging technology; Laboratories; Maps; Marketing; Methods; Microscopy; Molecular; National Institute of General Medical Sciences; Optics; Performance; Persons; Phase; Preparation; Process; Relative (related person); Research; Research Personnel; Sampling; Science; Solid; Specimen; Staging; Structure; Technology; Time; Universities; Work; base; bioimaging; cellular imaging; cost; electron tomography; imaging modality; improved; light microscopy; meetings; molecular imaging; molecular/cellular imaging; novel; professor; prototype; public health relevance; single molecule; three-dimensional modeling; tomography

DESCRIPTION (provided by applicant): Improved supporting technologies for imaging of molecular and supramolecular structures within cells are needed to facilitate cell biology research, and are sought by the National Institute of General Medical Sciences (specifically, its Division of Cell Biology and Biophysics). Multiscale imaging, using cryo-electron tomography (cryo-ET) on supramolecular structures and single molecules, has proven in recent years to be a unique and invaluable method for high-throughput characterization of the dynamic 3D architecture of cells. Electron microscopy (EM) grids, used as substrates for supporting the biological and biomolecular specimens being imaged, are a critical component associated with this imaging method. New EM grid technology that decreases sample preparation cost and time, improves sample generation from culturing to freezing for cryo-ET, and increases imaging quality will allow researchers to more efficiently explore cellular architecture, at higher throughput. Synkera proposes a novel ceramic EM grid that features an integrated thin support film that is highly compatible with cell culturing, light microscopy and cryo-ET. The grids will facilitate high-throughput, multiscale imaging of sub-cellular architecture and offer key advantages over state-of-the art products. The grids are also expected to be a competitive alternative in many other EM and culturing applications. Phase I work will demonstrate feasibility of the proposed fabrication method, based on micromachined nanoporous ceramic. Phase I will also demonstrate compatibility of the proposed EM grids with cell culturing, light microscopy and cryo-EM. EM grids will be fabricated with two different integrated support film options. These grids will be compared to traditional EM grid products (gold grids with holey carbon thin film supports) in order to demonstrate greater performance for multiscale cellular and molecular imaging. At least three academic partners will aide in demonstrating these capabilities. PUBLIC HEALTH RELEVANCE: The project addresses imaging of molecules and cells via cryo-electron tomography (cryo-ET). Specifically, the target application is multiscale imaging via optical microscopy and cryo-ET of cellular, supramolecular and single-molecule structures, for generating 3D models of sub-cellular architecture. The development of a novel class of ceramic-based electron microscopy grids that facilitate this multiscale imaging is proposed. The proposed technology will offer greater capability over state-of-the-art products and help further streamline multiscale cellular imaging by simplifying the specimen preparation process and yielding superior imaging performance.

描述（由申请人提供）：需要改进的细胞内分子和超分子结构成像的支持技术来促进细胞生物学研究，并且是国家普通医学科学研究所（特别是其细胞生物学和生物物理学部门）寻求的。近年来，使用冷冻电子断层扫描 (cryo-ET) 对超分子结构和单分子进行多尺度成像已被证明是对细胞动态 3D 结构进行高通量表征的独特且宝贵的方法。电子显微镜 (EM) 网格用作支撑正在成像的生物和生物分子样本的基底，是与该成像方法相关的关键组件。新的 EM 网格技术可降低样品制备成本和时间，改善冷冻 ET 从培养到冷冻的样品生成，并提高成像质量，将使研究人员能够以更高的通量更有效地探索细胞结构。 Synkera 提出了一种新型陶瓷 EM 网格，其具有集成薄支撑膜，与细胞培养、光学显微镜和冷冻电子断层高度兼容。该网格将促进亚细胞结构的高通量、多尺度成像，并提供优于最先进产品的关键优势。该网格也有望成为许多其他电磁和培养应用中具有竞争力的替代品。第一阶段的工作将证明所提出的基于微机械纳米多孔陶瓷的制造方法的可行性。第一阶段还将展示所提出的 EM 网格与细胞培养、光学显微镜和冷冻电镜的兼容性。电磁网格将采用两种不同的集成支撑膜选项来制造。这些网格将与传统的 EM 网格产品（带有多孔碳薄膜支撑的金网格）进行比较，以展示多尺度细胞和分子成像的更高性能。至少三个学术合作伙伴将协助展示这些能力。 公共健康相关性：该项目致力于通过冷冻电子断层扫描 (cryo-ET) 对分子和细胞进行成像。具体来说，目标应用是通过光学显微镜和冷冻电子断层扫描对细胞、超分子和单分子结构进行多尺度成像，以生成亚细胞结构的 3D 模型。提出了开发一种新型陶瓷基电子显微镜网格，以促进这种多尺度成像。所提出的技术将提供比最先进产品更强大的功能，并通过简化样本制备过程和产生卓越的成像性能，帮助进一步简化多尺度细胞成像。