TR&D Project 2: Tissue and cellular elemental distribution, and image correlation

项目概要- TR&D项目2 组织和细胞元素分布和图像相关性 本技术研究开发项目将开发扫描X射线荧光显微镜 （SFXM）用于定量观察细胞和组织中的微量元素分布。它将使用低温标本 制备和成像条件作为化学和结构保存的金标准， 抗辐射损伤我们将开发一种低温SXFM能力， 阿贡国家实验室先进光子源（APS）光束线8-BM的样本区，以及 支持其运作。这将使APS的低温SXFM实验时间增加一倍， 我们的BTRR和其他生物医学研究人员之间通过免费，同行评审的一般用户 向APS提出建议。 我们还将介绍与NIH的纳米级SFXM功能的冷冻标本交叉兼容性， 购买的Bionanoprobe在APS，激光消融电感耦合等离子体质谱（LA-ICP- MS; TR&D 1）在西北，光声显微镜（PAM; TR&D 2）在西北，和相关的 低温共聚焦光学显微镜（C3 LM）在APS。 我们将开发并提供PyElements，这是一个软件平台，提供一个集成和交叉的 来自所有这些模态的定量图像的配准视图。我们将在PyElements中加入一个 数值优化方法来校正较厚样品中X射线荧光的自吸收， 即使从较厚的3D样本中也可以获得定量信息。所有这些发展都将 发生在我们的驱动生物医学项目的背景下，特别是大面积低温SXFM 重要性主题A（金属稳态或脑功能失调），主题C（金属通量控制 生殖和发育）和主题D（代谢病理学中的金属失衡）。

PROJECT SUMMARY - TR&D PROJECT 2 Tissue and Cellular Elemental Distribution, and Image Correlation This Technology Research and Development Project will develop Scanning X-Ray Fluorescence Microscopy (SFXM) for quantitative views of trace element distributions in cells and tissues. It will use cryogenic specimen preparation and imaging conditions as a gold standard for chemical and structural preservation as well as radiation damage resistance. We will develop a cryo SXFM capability with a hundredfold increase in accessible specimen area for beamline 8-BM of the Advanced Photon Source (APS) at Argonne National Laboratory, and provide support for its operation. This will double the accessible experimental time in cryo SXFM at the APS, divided between our BTRR and other biomedical researchers via no-cost, peer-reviewed General User Proposals to the APS. We will also introduce cryo specimen cross-compatibility with the nanoscale SFXM capabilities of the NIH- purchased Bionanoprobe at the APS, laser-ablation inductively coupled plasma mass spectrometry (LA-ICP- MS; TR&D 1) at Northwestern, photoacoustic microscopy (PAM; TR&D 2) at Northwestern, and correlative cryogenic confocal light microscope (C3LM) at the APS. We will develop and make available PyElements, a software platform to provide an integrated and cross- registered view of quantitative images from all of these modalities. We will incorporate into PyElements a numerical optimization approach to correcting for self-absorption of x-ray fluorescence in thicker specimens, so that one can obtain quantitative information even from thicker specimens in 3D. All of these developments will take place in the context of our Driving Biomedical Projects, with large-area cryo SXFM being of particular importance to Theme A (Metal homeostasis or dysregulation in brain function), Theme C (Metal fluxes controlling reproduction and development), and Theme D (Metal imbalances in metabolic pathology).