Biological Soft X-ray Tomography at the Advanced Light Source

项目概要/摘要 国家X射线断层扫描中心（NCXT）是一个提供全方位服务的用户设施， 生物医学成像和研究。NCXT用户，无论是在现场还是远程工作， 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）和超分辨率低温荧光 断层扫描（SR-CFT）。在美国，这些成像模式仅在 NCXT。 SXT显微镜可视化和量化生物细胞的中尺度组织， 包括大的真核细胞。SXT不需要染色剂或造影剂 显像在被NCXT的透射软X射线显微镜成像之前， 先进的光源，细胞被快速冷冻保存;唯一的预成像步骤 必需的. SXT与其他成像方式，特别是荧光成像， 电子显微镜中尺度，整个细胞的3D SXT重建充当 用于解释其他类型的成像和生物物理数据的框架， 使它们可以在整个细胞的背景下被观察。 XM 2是目前国内唯一的显微镜。P30请求 资金将支持XM-2的日常运作，以及相关的 用户程序和标本制备实验室，以及我们的升级计划 软件/计算资源。在拨款周期内，我们会达致以下目标 具体目标： 目标1：为生物医学研究界提供软X射线 断层扫描（SXT）。这一目标的工作确保了SXT的细胞成像仍然是一个有效的方法。 向美国研究人员开放。将认真考虑使 资源尽可能方便用户，特别是对研究人员社区 远程工作 目的2：维护和校准NCXT操作的XM-2软X射线机 显微镜在ALS与所有的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。我们就来 确保XM-2坚固、可靠并以最小的 计划外停机时间。 目标3：实施升级和增强，以确保NCXT 仍然是“最先进的”。在融资周期的时间范围内，新的硬件， 软件和算法将变得可用，一些现有的组件将 到达他们的生命终点。因此，我们将进行升级和改进， 将NCXT保持在最先进的水平。'

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.'