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

项目摘要/摘要 X射线显微镜已经发展成为一种重要的新的细胞成像技术 三维可视化和测量完整的细胞。这些系统的发展， 在X射线光学和探测器技术进步的推动下，主要是在 同步辐射光源设施。第一台用于细胞成像的实验室X射线显微镜 由提案的联合PI在2009年引入，并利用吸收对比 用于成像的“窗口”(285-540 eV)X射线。该系统实现了未染色的细胞断层扫描 这是第一次在实验室里。然而，对于大多数人来说，系统运行能量太低 哺乳动物细胞，其直径大于真核细胞(酵母、细菌等)。细胞 水窗能使。 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FAST(~30 分钟)和高对比度完整的直径达80um的未染色细胞的3D成像 它们的细胞器的分辨率为30 nm。该系统使用Zernike相位对比度在 2.7kev能量x射线，其对比度高于水的均匀吸收对比度 窗口X光片。此外，与水相比，该系统将具有几个主要优势 窗口X射线显微镜，包括：更大的细胞成像(80微米对10微米)，更大的深度- 更高的3D分辨率和实用的优势(更稳定的x射线源和更大的 工作距离以纳入相关技术)。显微镜使用的是该公司的 获得专利的高亮度X射线源和专有的X射线光学技术。 拟议的第二阶段为期24个月的项目是开发2.7kev泽尼克的完整原型 用于细胞成像的相位对比度系统，并通过实验验证其性能。

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.