Cryogenic High-throughput Cellular Imaging System

低温高通量细胞成像系统

• 批准号: 10545696
• 负责人: Wenbing Yun
• 金额: $ 99.8万
• 依托单位: SIGRAY, INC.
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-10 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10545696
• 关键词: 3-Dimensional; Address; Algae; Attenuated; Bacteria; Biological; Caliber; Carbon; Cell Communication; Cell Size; Cells; Cryoelectron Microscopy; Cryopreservation; Development; Disease; Dose; Electron Microscopy; Electrons; Freezing; Gases; Germany; Grant; Hela Cells; Hydration status; Ice; Image; Laboratories; Legal patent; Letters; Light; Link; M cell; Mammalian Cell; Measures; Membrane; Microscope; Nitrogen; Optics; Organelles; Oxygen; Performance; Phase; Preparation; Radiation induced damage; Resolution; Roentgen Rays; Sampling; Small Business Innovation Research Grant; Source; Specimen; Stains; Stream; Structure; Synchrotrons; System; Taiwan; Techniques; Technology; Three-Dimensional Imaging; United States National Institutes of Health; Water; X ray microscopy; Yeasts; absorption; beamline; biological research; cellular imaging; cryogenics; design; detection platform; high resolution imaging; imaging approach; imaging capabilities; imaging system; innovation; light microscopy; microscopic imaging; nano; nanometer resolution; operation; preservation; radiation effect; tomography; tool; transmission process; water sampling

Project Summary/Abstract X-ray microscopy has evolved into an important ultrastructure imaging approach for visualizing and measuring intact cells in three dimensions. Beamlines dedicated to cellular imaging have been constructed at synchrotron facilities around the world, and the co-PI of this proposal developed the first commercial laboratory x-ray microscope for cellular imaging. The approaches thus far have centered around the use of “water window” x-rays, having energies between 285 to 540eV and for which water is transparent but organic content is absorbing. However, there are several major drawbacks to using these low energy x-rays: they severely limit the size of cells that can be imaged (e.g., <10 µm when many mammalian cells are 10-100 µm). This SBIR proposal aims to develop a 3D nano x-ray microscope capable of high throughput (~30 minute) 3D imaging of cryogenically preserved cells of up to 80 µm in diameter at down to 30nm resolution. The system utilizes the phenomenon of Zernike phase contrast at higher (2.7 keV) energy x-rays, which can achieve even higher contrast for biological samples than water window x-rays. Additionally, the system will enable several major advantages over water window x-ray microscopy, including much larger cell imaging (80µm vs. 10µm), larger depth-of- field for higher 3D resolution, and practical benefits (more stable x-ray source and larger working distance to incorporate correlative techniques). The microscope uses the company’s patented high brightness x-ray source and proprietary x-ray optic technology. The project will develop the proposing company’s existing 2.7 keV ambient system to enable cryogenic operation and optimize its performance for cellular imaging. The proposed Phase II 24-month project is to develop a complete cryogenic 2.7 keV system for cellular imaging and to experimentally demonstrate its performance on mammalian cells.

项目摘要/摘要 X射线显微镜已演变为一种重要的超微结构成像方法，以可视化 并在三个维度上测量完整的细胞。专用于细胞成像的光束线具有 是在世界各地的同步设施中建造的，该提议的共同点 开发了第一个用于细胞成像的商业实验室X射线显微镜。这 迄今 在285至540EV之间，水是透明的，但有机含量正在吸收。 但是，使用这些低能X射线有几个主要缺点：它们严重限制了 可以成像的细胞大小（例如，当许多哺乳动物细胞为10-100 µm时，<10 µm）。 该SBIR建议旨在开发能够高通量的3D纳米X射线显微镜 （〜30分钟）直径高达80 µm的低温保存细胞的3D成像 30nm分辨率。该系统利用Zernike阶段对比的现象在较高的情况下（2.7 Kev）Energy X射线，该射线可以比水获得更高的生物样品对比度 窗口X射线。此外，该系统将在水中实现几个主要优势 窗户X射线显微镜，包括更大的单元成像（80µm vs. 10µm），更大的深度 更高3D分辨率和实际收益的领域（更稳定的X射线源和更大的领域 加入相关技术的工作距离）。显微镜使用公司的 获得专利的高亮度X射线源和专有X射线光学技术。 该项目将开发提案公司现有的2.7 KEV环境系统，以启用 低温操作并优化其用于细胞成像的性能。提出的II期 24个月的项目是开发一个完整的低温2.7 KEV系统，用于细胞成像和至 实验表明其在哺乳动物细胞上的性能。