Biological Soft X-ray Tomography at the Advanced Light Source

项目摘要/摘要 国家X射线层析成像中心(NCXT)是一个全方位服务的用户设施，用于 生物医学成像和研究。NCXT用户，无论是现场工作还是远程工作， 能够使用专门的显微镜和成像技术，特别是软x射线 层析成像(SXT)和超分辨低温荧光 断层扫描(SR-CFT)。在美国，这些成像设备只在 NCXT。 SXT显微镜可视化和量化生物细胞的中尺度组织， 包括大型真核细胞。SXT不需要染色或造影剂 成像。在被NCXT的透射式软X射线显微镜在 先进的光源，细胞被快速冷冻保存；唯一的成像前步骤 必填项。 SXT与其他成像方式有很强的互补性，特别是荧光和 电子显微镜。整个细胞的中尺度3D SXT重建充当 解释其他类型的成像和生物物理数据的框架， 从而允许在整个单元的上下文中查看它们。 XM2是目前国内唯一的此类显微镜。请求的P30 资金将支持XM-2的日常运作，以及相关的 用户程序和样品制备实验室，以及升级到我们的 软件/计算资源。在筹资周期内，我们将实现以下目标 具体目标： 目标1：让生物医学研究团体获得软x射线 断层扫描(SXT)。这一目标的工作确保了SXT的成像细胞仍然是 美国研究人员可以选择。我们将认真考虑制定 尽可能方便用户使用的资源，特别是针对研究人员社区 它可以远程工作。 目的2：维修和校准NCXT操作的软X射线XM-2 肌萎缩侧索硬化症的显微镜。与所有同步加速器光束线一样，XM-2是一种复杂的 需要定期维护和校准的仪器。我们将采取一项 确保XM-2健壮、可靠并以最低限度运行的程序化方法 计划外停机。 目标3：实施升级和增强以确保NCXT 仍然是最先进的。在资金周期的时间范围内，新的硬件， 软件和算法将可用，一些现有组件将 到达他们生命的尽头。因此，我们将进行升级和增强，以 保持NCXT处于最先进的水平。

Project Summary/Abstract The National Center for X-ray Tomography (NCXT) is a full-service, user facility for biomedical imaging and research. NCXT Users, whether working on site or remotely, have access to specialized microscopes and imaging techniques, in particular soft x-ray tomography (SXT) and super-resolution cryogenic fluorescence tomography (SR-CFT). In the US, these imaging modalities are only available at the NCXT. SXT microscopes visualize and quantify the mesoscale organization of biological cells, including large eukaryotic cells. No stains or contrast agents are required for SXT imaging. Before being imaged by the NCXT's transmission soft x-ray microscope at the Advanced Light Source, cells are rapidly cryo-preserved; the only pre-imaging step required. SXT is highly complementary to other imaging modalities, particularly fluorescence and electron microscopies. Mesoscale, 3D SXT reconstructions of an entire cell act as a framework for the interpretation of other types of imaging and biophysical data, allowing them to be viewed in the context of the whole cell. XM2 is currently the only microscope of its kind in the Nation. The requested P30 funding will support the day-to-day operation of XM-2, together with the associated user program and specimen preparation laboratories, and a program of upgrades to our software/computing resources. During the funding cycle, we will achieve the following specific aims: Aim 1: Give the biomedical research community access to soft x-ray tomography (SXT). The work of this aim ensures imaging cells by SXT remains an option open to US researchers. Significant consideration will be given to making the Resource as user-friendly as possible, particular towards the community of researchers that work remotely. Aim 2: Maintain and calibrate XM-2, the NCXT operated soft x-ray microscope at the ALS. As with all synchrotron beamlines, XM-2 is a sophisticated instrument that requires regular maintenance and calibration. We will take a programmatic approach to ensure XM-2 is robust, reliable, and operates with minimum unscheduled downtime. Aim 3: Implement upgrades and enhancements to ensure the NCXT remains `state-of-the-art.' Over the timescale of the funding cycle, new hardware, software, and algorithms will become available, and some existing components will reach their end-of-life. We will, therefore, carry out upgrades and enhancements to maintain the NCXT at the `state-of-the-art.'