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

获得这一奖项的是，化学部生命过程化学项目支持佐治亚理工学院的Christoph J.Fahrni博士建立了一种多模式成像方法，用于研究斑马鱼胚胎发育中锌池和增殖活动区域的空间相关性。拟议工作中使用的成像方式代表了当前痕量金属成像研究的前沿。使用一系列经过特殊优化的发射比率荧光锌探针，用双光子激发显微镜(TPEM)显示了活体胚胎中不稳定锌库的三维再分布动力学。同时，利用转基因斑马鱼品系(ZFucci)鉴定有丝分裂活动区，在该品系中，增殖细胞表达荧光标记蛋白。在选定的发育阶段，最近在阿贡国家实验室的高级光子源实施的XRF断层扫描也显示了锌的总分布。这种多模式成像方法将首次提供对锌库重新分布动态的直接洞察，以及它们在早期胚胎发育过程中的细胞周期进程中的作用。锌是所有生命有机体活力所必需的微量营养物质。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，也是正常胚胎发育的关键。由于不同器官对锌的需求差异很大，胚胎很可能在发育过程中被迫重新分配有限的微量营养物质。尽管锌在发育过程中的调控和再分配机制具有根本性的重要性，但在很大程度上仍未被探索。进展一直受到阻碍，部分原因是与量化现场锌的分布有关的挑战。这三种成像模式的结合将允许在空间上关联单个胚胎中不稳定锌库的位置、总锌的分布和细胞周期进程，从而阐明锌在胚胎发生过程中的增殖活动中的作用。多模式成像方法有望很容易地适用于研究与发育过程中锌稳态有关的广泛问题。Fahrni博士还将与佐治亚州Marietta的Wheeler High School的John Cody博士合作，为学生提供参加为期四周的暑期项目的机会，该项目将让他们体验一系列尖端研究技术，包括合成化学、光谱学、光物理和生物成像。预计今年暑期项目将提高学生在STEM相关领域继续学习的热情。该奖项由生物科学(BIO)理事会综合组织系统(IOS)部门的生理和结构系统(PSS)集群共同资助。

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.