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

项目概要-研发项目1 高通量多元素分布和组织水平定量 金属对于具有高度调节的运输、定位和稳态的生理过程至关重要， 其中中断或轻微的变化导致细胞和组织功能障碍并最终导致疾病。这 技术研究与发展（TR&D）项目将推进激光烧蚀感应耦合等离子体 飞行时间质谱（LA-ICP-TOFMS）技术，以开发更高通量的成像 在不同的生物医学应用中的金属容量。这一办法将补充正在制定的办法 在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的作用有一个全面的了解。 使用我们的驾驶生物医学项目（DBP）作为试验台，该TR&D将集成LA-ICP-TOFMS组件 开发用于获取从独特组织类型导出的快速定量2D多元素标测图的工作流程 不同的样品制备方法。使用低温LA池将能够低温分析 保存样本，确保组织化学和结构得到保存。我们还将制定标准化的 生产组织特异性LA-ICP-MS校准标准品的方法。我们的标准将得到验证 在光束线8-BM使用SXFM。一旦制定了标准生产和验证的协议， 大量组织特异性SXFM验证标准品将作为通用标准品提供给社区 外部LA-ICP-MS校准标准品。最后，由于较大体积的金属热点可能是 在2D图像中被歪曲或遗漏，我们将开发3D成像LA-ICP-TOFMS技术， 器官和有机体。为了生成3D图像，我们将使用由快速切片技术促进的连续切片技术。 TOFMS的图像采集能力，并结合多模态图像分析软件 （PyElements）在TR&D 2中开发。3D成像功能将支持更广泛的适用性， 生物成像技术为生物医学界。所有这些事态发展都将发生在 我们的DBPs，从所有四个主题的项目（金属稳态或脑功能失调;金属 调节宿主-病原体相互作用;控制生殖和发育的金属通量;以及 代谢病理学失衡），支持组织特异性校准标准的开发， 用于分析不同组织类型的2D切片的工作流程。LA-ICP-TOFMS三维成像技术的研究进展 技术将与DBP主题C（控制生殖和发展的金属通量）最为相关。 一旦成熟，低温-LA-ICP-TOFMS将被纳入西北大学的定量 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.