Ceramic Electron Microscopy Grids for Cell Culturing and Multiscale Imaging

描述（由申请人提供）：需要改进细胞内分子和超分子结构成像的支持技术，以促进细胞生物学研究，并且国立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）正在寻求这种技术。 多尺度成像，使用冷冻电子断层扫描（cryo-ET）的超分子结构和单分子，近年来已被证明是一个独特的和宝贵的方法，用于高通量表征的动态三维结构的细胞。电子显微镜（EM）的网格，作为支持生物和生物分子样本被成像的基板，是与这种成像方法相关的关键组成部分。新的EM网格技术降低了样品制备成本和时间，改善了从培养到冷冻的冷冻ET样品的生成，并提高了成像质量，这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提出了一类新型陶瓷EM网格，其特征在于集成薄支撑膜，与细胞培养、光学显微镜和冷冻ET高度兼容。该网格将促进亚细胞架构的高通量、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EM和培养应用的竞争性替代品，一个为期6个月的第一阶段项目成功证明了拟议网格的可行性。 结构，以及它们在培养、冷冻ET和多尺度成像中的潜力。第二阶段的目标是通过进一步开发制造工艺和陶瓷EM网格设计来建立这一成功，以充分实现功能原型中多尺度成像的潜力。至少有四个学术合作伙伴将协助这一发展过程。拟议项目的最终目标是为广泛的EM应用提供一系列完整的陶瓷EM网格，从生物成像到材料表征。

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 highthroughput 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 class of ceramic EM grids that feature 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 a 6-month Phase I project successfully demonstrated feasibility of the proposed grids architecture, as well as their potential in culturing, cryo-ET and multiscale imaging. Phase II aims to build on this success by further developing fabrication processes and ceramic EM grids designs, to fully realize the potential for multiscale imaging in functional prototypes. At least four academic partners will aide in this development process. The ultimate goal of the proposed project is a complete line of ceramic EM grids for a broad range of EM applications, from bioimaging to materials characterization.