Development of a Laboratory Transmission X-ray Microscope for Rapid 3D Cellular Tomography using Zernike Phase Contrast with 2.7 keV

使用 2.7 keV 泽尼克相差技术开发用于快速 3D 细胞断层扫描的实验室透射 X 射线显微镜

• 批准号: 10011500
• 负责人: David Vine
• 金额: $ 85.77万
• 依托单位: SIGRAY, INC.
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-15 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10011500
• 关键词: 3-Dimensional; Algae; Anodes; Area; Attenuated; Bacteria; Biological; Biology; Caliber; Carbon; Cell Nucleus; Cell physiology; Cells; Cellular Structures; Complement; Cryopreserved Cell; DNA; Development; Diagnostic radiologic examination; Diamond; Disease; Electrons; Fluorescence Microscopy; Hydration status; Image; Imaging Techniques; Laboratories; Legal patent; Light; Link; Mammalian Cell; Measures; Medical; Methods; Microscope; Microscopy; Molecular; Morphologic artifacts; Mutation; Operating System; Optics; Organelles; Oxygen; Performance; Phase; Photons; Proxy; RNA; Research; Resolution; Roentgen Rays; Science; Small Business Innovation Research Grant; Solid; Source; Stains; Structure; Synchrotrons; System; Systems Development; Techniques; Technology; Three-Dimensional Image; Three-Dimensional Imaging; Time; Water; X ray microscopy; Yeasts; absorption; base; cell water; cellular development; cellular imaging; contrast imaging; design; detector; high resolution imaging; imaging capabilities; imaging modality; improved; innovation; operation; preservation; prototype; screening; statistics; tomography; transmission process

Project Summary/Abstract X-ray microscopy has newly developed into an important new cellular imaging technique for visualizing and measuring intact cells in three dimensions. The development of these systems, enabled by advances in x-ray optic and detector technology, has been primarily driven at synchrotron light source facilities. The first laboratory x-ray microscopes for cellular imaging were introduced by the proposal’s co-PI in 2009 and utilized the absorption contrast of “water window” (285-540eV) x-rays for imaging. The system enabled unstained cellular tomography for the first time in the laboratory. However, the system operational energy is too low for most mammalian cells, which are larger in diameter than eukaryotic (yeast, bacteria, etc.) cells that the water window enables. We propose to develop a new 2.7 keV x-ray laboratory cellular microscope to provide fast (~30 minute) and high contrast complete 3D imaging of unstained cells of up to 80um in diameter and their organelles at 30nm resolution. The system operates using Zernike phase contrast at 2.7 keV energy x-rays, which provides higher contrast than even absorption contrast of 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 proposed Phase II 24-month project is to develop a complete prototype 2.7 keV Zernike phase contrast system for cellular imaging and to experimentally verify its performance.

项目总结/摘要 X射线显微镜已发展成为一种重要的新的细胞成像技术， 可视化和测量三维的完整细胞。这些系统的发展， 由于X射线光学和探测器技术的进步， 同步加速器光源设施。第一台用于细胞成像的实验室X射线显微镜 2009年，该提案的共同提案人介绍了这一技术，并利用了“水”的吸收对比度 窗口”（285- 540 eV）X射线成像。该系统能够进行未染色的细胞断层扫描 这是第一次在实验室中进行。然而，系统运行能量对于大多数 哺乳动物细胞，其直径大于真核细胞（酵母、细菌等）。的细胞 水窗可以。 我们建议开发一种新的2.7keV X射线实验室细胞显微镜，以提供快速（~30 分钟）和高对比度完整的3D成像，直径高达80 μ m的未染色细胞 和它们的细胞器。该系统使用Zernike相位衬度操作， 2.7 keV能量x射线，其提供比水的均匀吸收对比度更高的对比度 窗户X光片此外，该系统将使几个主要优势超过水 窗口X射线显微镜，包括：更大的细胞成像（80µm vs. 10µm），更大的深度- 更高的3D分辨率和实际好处（更稳定的X射线源和更大的 工作距离，以结合相关技术）。显微镜使用该公司的 获得专利的高亮度x射线源和专有的x射线光学技术。 拟议的第二阶段24个月的项目是开发一个完整的原型2.7 keV泽尼克 相衬系统的细胞成像和实验验证其性能。