TR&D Project 1: Higher Throughput Multi-element Distribution & Quantitation at the Tissue Level

项目摘要 – TR&D 项目 1 组织水平上更高通量的多元素分布和定量 金属对于高度调控的运输、定位和体内平衡的生理过程至关重要， 中断或轻微的变化会导致细胞和组织功能障碍并最终导致疾病。这个 技术研发（TR&D）项目将推进激光烧蚀电感耦合等离子体 用于开发更高通量成像的飞行时间质谱 (LA-ICP-TOFMS) 技术 金属在多种生物医学应用中的能力。这种方法将补充那些正在开发的方法 TR&D 2 和 3 中，使生物医学研究人员能够平衡灵敏度、选择性和空间分辨率 需要对金属在基本生物过程中发挥的作用有一个全面的了解。 使用我们的驱动生物医学项目 (DBP) 作为试验台，该 TR&D 将集成 LA-ICP-TOFMS 组件 开发用于获取源自独特组织类型的快速定量二维多元素图的工作流程 以及不同的样品制备方法。使用低温 LA 细胞将能够进行低温分析 保存样本，确保组织化学和结构得到保留。我们还将制定标准化 生产组织特异性 LA-ICP-MS 校准标准的方法。我们的标准将得到验证 在光束线 8-BM 处使用 SXFM。一旦制定了标准生产和验证协议，就会有一套 一定数量的组织特定 SXFM 验证标准将作为通用标准提供给社区 外部 LA-ICP-MS 校准标准。最后，因为较大体积的金属热点可以 2D 图像中的误报或遗漏，我们将开发 3D 成像 LA-ICP-TOFMS 技术来解决整个问题 器官和生物体。为了生成 3D 图像，我们将使用由快速 TOFMS 的图像采集功能并结合多模态图像分析软件 (PyElements) 正在 TR&D 2 中开发。3D 成像功能将支持更广泛的适用性 生物医学界的生物成像技术。所有这些发展都将在这样的背景下发生 我们的 DBP，涉及所有四个主题的项目（金属稳态或脑功能失调；金属 宿主-病原体相互作用的调节；控制繁殖和发育的金属通量；和金属 代谢病理学的不平衡），支持组织特异性校准标准的开发和 用于分析不同组织类型的 2D 切片的工作流程。 3D LA-ICP-TOFMS 成像的开发 技术将与 DBP 主题 C（控制繁殖和发育的金属通量）最相关。 一旦成熟，cryo-LA-ICP-TOFMS 将作为服务线纳入西北大学的定量分析中心 生物元素成像中心。

PROJECT SUMMARY – TR&D PROJECT 1 Higher Throughput Multi-element Distributions & Quantitation at the Tissue Level Metals are crucial for physiological processes with highly regulated trafficking, localization, and homeostasis, where interruptions or slight variations lead to cell and tissue dysfunction and ultimately to disease. This technology research & development (TR&D) project will advance laser ablation inductively coupled plasma time-of-flight mass spectrometry (LA-ICP-TOFMS) techniques to develop increased throughput imaging capacity for metals in diverse biomedical applications. This approach will complement those being developed in TR&Ds 2 and 3, enabling biomedical researchers to balance sensitivity, selectivity, and spatial resolution needs to develop a wholistic understanding of the role metals play in fundamental biological processes. Using our Driving Biomedical Projects (DBP) as test beds, this TR&D will integrate LA-ICP-TOFMS components to develop workflows for acquiring rapid quantitative 2D multi-elemental maps derived from unique tissue types and different sample preparation methods. Use of a cryogenic LA cell will enable analysis of cryogenically preserved samples, ensuring tissue chemistry and structure are preserved. We will also develop standardized approaches for producing tissue-specific LA-ICP-MS calibration standards. Our standards will be validated using SXFM at beamline 8-BM. Once protocols for standard production and validation are developed, a set quantity of tissue specific SXFM-validated standards will be made available to the community as universal external LA-ICP-MS calibration standards. Finally, because metal hotspots in larger volumes can be misrepresented or missed in 2D images, we will develop 3D imaging LA-ICP-TOFMS techniques for whole organs and organisms. To produce 3D images we will use serial sectioning techniques facilitated by the rapid image acquisition capabilities of the TOFMS and incorporate multi-modality image analysis software (PyElements) being developed in TR&D 2. 3D imaging capabilities will support a wider applicability of bioimaging techniques for the biomedical community. All of these developments will take place in the context of our DBPs, with projects from all four themes (Metal homeostasis or dysregulation in brain function; metal modulation in host-pathogen interactions; metal fluxes controlling reproduction and development; and metal imbalances in metabolic pathology), supporting development of tissue-specific calibration standards and workflows for analyzing 2D sections of different tissue types. Development of 3D LA-ICP-TOFMS imaging technology will be most relevant to DBP Theme C (metal fluxes controlling reproduction and development). Once mature, the cryo-LA-ICP-TOFMS will be incorporated as a service line in Northwestern’s Quantitative Bioelement Imaging Center.