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

项目摘要-研发项目1 更高吞吐量的多元素分布和组织水平的定量 金属对具有高度调控的运输、定位和动态平衡的生理过程至关重要， 干扰或轻微变异会导致细胞和组织功能障碍，并最终导致疾病。这 技术研发项目将推动激光烧蚀感应耦合等离子体 飞行时间质谱学(LA-ICP-TOFMS)技术用于开发更高吞吐量的成像 金属在不同的生物医学应用中的能力。这种方法将对正在开发的方法进行补充 在R&D 2和3中，使生物医学研究人员能够平衡敏感度、选择性和空间分辨率 需要对金属在基本的生物过程中所起的作用有一个整体的理解。 使用我们的驾驶生物医学项目(DBP)作为试验台，该研发将集成LA-ICP-TOFMS组件 开发从独特组织类型获取快速定量2D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池将能够进行低温分析 保存的样本，确保组织化学和结构得到保存。我们还将制定标准化的 组织特异性LA-ICPMS校准标准品的制备方法。我们的标准将得到验证 在光束线8-BM使用SXFM。一旦制定了用于标准生产和验证的协议，一套 将向社区提供大量组织特定的SXFM验证标准作为通用标准 外部LA-ICPMS校准标准。最后，因为体积较大的金属热点可能是 在2D图像中被错误描述或遗漏的，我们将开发3D成像LA-ICP-TOFMS技术 器官和生物体。为了生成3D图像，我们将使用由Rapid帮助的连续切片技术 TOFMS的图像采集能力，并整合了多通道图像分析软件 (PyElements)正在研发中。3D成像功能将支持更广泛的适用性 生物医学领域的生物成像技术。所有这些发展都将在 所有四个主题的项目(金属稳态或大脑功能失调；金属 寄主-病原体相互作用的调节；控制繁殖和发育的金属熔剂；以及金属 代谢病理学中的不平衡)，支持制定组织特异性校准标准和 用于分析不同组织类型的二维切片的工作流。三维LA-ICP-TOFMS成像技术的研究进展 技术将与DBP主题C(控制繁殖和发育的金属熔剂)最为相关。 一旦成熟，Cryo-LA-ICP-TOFMS将作为一条服务线纳入西北大学的量化 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.