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

TR

• 批准号: 10197969
• 负责人: THOMAS V O'HALLORAN
• 金额: $ 28.23万
• 依托单位: MICHIGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10197969
• 关键词: 3-Dimensional; Ablation; Adopted; Automobile Driving; Beds; Biological; Biological Preservation; Biological Process; Biological Sciences; Brain; Calibration; Cells; Chemistry; Communities; Complement; Complex; Computer software; Coupled; Crustacea; Data; Data Analyses; Data Set; Development; Disease; Elements; Ensure; Equilibrium; Fresh Water; Functional disorder; Goals; Grain; Heterogeneity; Homeostasis; Hour; Image; Image Analysis; Imaging Techniques; Imaging technology; Individual; Inductively Coupled Plasma Mass Spectrometry; Institution; Interruption; Lasers; Lead; Left; Link; Maps; Mass Spectrum Analysis; Metabolic; Metals; Methods; Multimodal Imaging; Mus; Organ; Organism; Pathology; Physiological Processes; Plasma; Play; Preparation; Procedures; Process; Production; Protocols documentation; Pythons; Reference Standards; Reporting; Reproduction; Research Personnel; Resolution; Resources; Risk; Role; Rye cereal; Sampling; Sectioning technique; Services; Specificity; Standardization; Structure; System; Systems Integration; Techniques; Testing; Three-Dimensional Image; Three-Dimensional Imaging; Time; Tissue Sample; Tissues; Validation; Variant; Visualization; Wheat; base; beamline; bioimaging; biological systems; cryogenics; data acquisition; data reduction; design; detection limit; detector; imaging approach; imaging capabilities; improved; open source; pathogen; photoacoustic imaging; preservation; spatiotemporal; technology research and development; time of flight mass spectrometry; tissue reconstruction; trafficking

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.

项目摘要 - TR＆D项目1 在组织水平上较高的吞吐量多元分布和定量 金属对于具有高度监管的贩运，本地化和体内平衡的物理过程至关重要， 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 技术研究与开发（TR＆D）项目将推动激光消融能力耦合等离子体 飞行时间质谱（LA-ICP-TOFMS）技术以开发增加的吞吐量成像 潜水员生物医学应用中金属的能力。这种方法将完成正在开发的方法 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学过程中所扮演的作用有全面的了解。 将我们的驾驶生物医学项目（DBP）作为测试床，该TR＆D将整合LA-ICP-TOFMS组件 开发工作流程，以辅助从独特组织类型得出的快速定量2D多元图 和不同的样品制备方法。使用低温LA细胞可以对低温分析 保留的样品，确保组织化学和结构保留。我们还将开发标准化 生成组织特异性LA-ICP-MS校准标准的方法。我们的标准将得到验证 在光线线8-BM处使用SXFM。一旦开发了标准生产和验证协议， 作为通用 外部LA-ICP-MS校准标准。最后，因为大量的金属热点可以是 在2D图像中陈述或错过了，我们将为整体开发3D成像LA-ICP-TOFMS技术 器官和生物。为了产生3D图像，我们将使用快速制备的串行切片技术 TOFM和合并多模式图像分析软件的图像采集功能 （pyelements）在TR＆D2。3D成像功能中开发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上下文中进行 我们的DBP的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；金属通量控制繁殖和发育；和金属 代谢病理的失衡），支持组织特异性校准标准和 用于分析不同组织类型的2D部分的工作流程。开发3D LA-ICP-TOFMS成像 技术将与DBP主题C（控制繁殖和开发的金属通量）最相关。 一旦成熟，Cryo-la-iCp-Tofms将被合并为西北定量的服务线 生物元素成像中心。