Correlative X-ray Fluorescence Tomography and Multiphoton Imaging of Zinc in Developing Zebrafish Embryos

斑马鱼胚胎发育中锌的相关 X 射线荧光断层扫描和多光子成像

• 批准号: 1306943
• 负责人: Christoph Fahrni
• 金额: $ 40.5万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-15 至 2018-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1306943&HistoricalAwards=false
• 关键词: Correlative; ray; Fluorescence; Tomography; Multiphoton

With this award, the Chemistry of Life Processes Program in the Chemistry Division supports Dr. Christoph J. Fahrni from the Georgia Institute of Technology to establish a multimodal imaging approach for the spatial correlation of Zn pools and areas of proliferating activities within developing zebrafish embryos. The imaging modalities utilized in the proposed work represent the forefront of current trace metal imaging research. Using a series of specifically optimized emission ratiometric fluorescent zinc probes, the 3D redistribution dynamics of labile zinc pools are visualized in live embryos by two-photon excitation microscopy (TPEM). In parallel, areas of mitotic activity are identified using a transgenic zebrafish line (zFucci), in which proliferating cells are expressing a fluorescent marker protein. At selected developmental stages, the total zinc distribution are also visualized by XRF tomography as recently implemented at the Advanced Photon Source of the Argonne National Laboratory. This multimodal imaging approach should provide for the first time direct insights into the redistribution dynamics of zinc pools and their role in the cell cycle progression during early embryogenesis.Zinc is an essential trace nutrient required for the vitality of all living organisms. It plays a central role in major cellular processes, and it is required for cell proliferation and critical to proper embryonic development. As the zinc requirements vary widely between organs, the embryo is most likely forced to redistribute the limited supply of trace nutrients in the course of development. Despite being of fundamental importance, the mechanisms governing zinc regulation and redistribution during development remain largely unexplored. Progress has been hampered in part due to the challenges associated with quantifying the distribution of zinc in situ. The combination of the three imaging modalities will allow to spatially correlate the location of labile Zn pools, the distribution of total Zn, and the cell-cycle progression, in a single embryo, and thus to elucidate the role of Zn in proliferating activities during embryogenesis. The multimodal imaging approach is expected to be readily adaptable for the investigation of a broad range of questions concerning Zn homeostasis during development. Dr. Fahrni will also work with Dr. John Cody at Wheeler High School in Marietta, Georgia to provide students with an opportunity to participate in a four-week summer program that would give them experience in a range of cutting-edge research techniques, including synthetic chemistry, spectroscopy, photophysics, and biological imaging. It is expected that this summer program will increase the enthusiasm of the students for further study in STEM-related fields.This award is being co-funded by the Physiological and Structural Systems (PSS) Cluster of the Integrative Organismal Systems (IOS) Division of the Biological Sciences (BIO) Directorate.

有了这个奖项，化学部的生命过程化学项目支持格鲁吉亚理工学院的Christoph J. Fahrni博士建立一种多模态成像方法，用于锌池的空间相关性和发育中的斑马鱼胚胎内的增殖活动区域。在拟议的工作中使用的成像方式代表了当前痕量金属成像研究的最前沿。使用一系列专门优化的发射率荧光锌探针，三维再分配动态的不稳定锌池可视化活胚胎的双光子激发显微镜（TPEM）。与此同时，有丝分裂活性的区域使用转基因斑马鱼系（zFucci）进行鉴定，其中增殖细胞表达荧光标记蛋白。在选定的发展阶段，总锌分布也可视化的XRF断层扫描，最近实施的先进光子源的阿贡国家实验室。这种多模态成像方法应提供第一次直接洞察锌池的再分配动态及其在早期胚胎发生过程中的细胞周期进程中的作用。锌是所有生物体活力所需的必需微量营养素。 它在主要的细胞过程中起着核心作用，是细胞增殖所必需的，对正常的胚胎发育至关重要。 由于锌的需求在器官之间差异很大，胚胎很可能被迫在发育过程中重新分配有限的微量营养素供应。 尽管具有根本的重要性，但在发育过程中锌的调节和再分配机制在很大程度上仍未被探索。 进展受到阻碍，部分原因是与量化锌的原位分布相关的挑战。 这三种成像方式的组合将允许空间相关的位置不稳定的锌池，总锌的分布，和细胞周期的进展，在一个单一的胚胎，从而阐明锌在胚胎发生过程中的增殖活动的作用。多模态成像方法预计将很容易适应调查的范围广泛的问题，锌稳态在发展过程中。 Fahrni博士还将与格鲁吉亚玛丽埃塔的惠勒高中的John Cody博士合作，为学生提供参加为期四周的暑期课程的机会，这将使他们在一系列尖端研究技术方面获得经验，包括合成化学，光谱学，生物物理学和生物成像。 预计这个暑期项目将提高学生在STEM相关领域进一步学习的热情。该奖项由生物科学（BIO）理事会综合有机系统（IOS）部门的生理和结构系统（PSS）集群共同资助。