IDBR: TYPE A - Development of an In situ Single-cell Mass Spectrometer for Mapping Small-molecule Expression in the Developing Embryo

乔治华盛顿大学被授予开发一种设备，该设备将能够测量在发育中的胚胎的多个单个细胞中产生广泛类型的生物分子。该项目将通过整合生物和化学来加强教育，并提供新的调查工具，以提高基础研究和应用研究的创造性研究机会。单细胞质谱仪的开发将需要分析化学家、生物学家、质谱学家和数据存储库馆长之间的定期互动，基本上为学生和研究人员创造一个跨学科的环境，以完成这些学科经典课程之外的培训。通过在乔治华盛顿大学展示该设备，并在国家会议和出版物上讨论该设备的设计、性能和使用，这项工作将扩大科学素养，并使更广泛的用户群了解该设备的可用性。胚胎发育过程中生物分子产生的测量数据将在公众可访问的数据储存库中传播，为更多的用户提供便利，促进细胞和发育生物学和神经科学的研究和教育。值得一提的是，这些科学和宣传元素的结合，包括当地高中生参与该项目，将加强生物学、仪器开发和分析化学方面的研究和教育，为科学和教育领域当前和未来的挑战提供跨学科的解决方案。胚胎单细胞生物分子表达的特征将为协调胚胎发育的基本生化机制提供新的见解，即受精卵通过一系列复杂的过程产生一个完整的、功能齐全的有机体，如青蛙、鱼或人类。虽然在单个胚胎细胞中测量基因和转录物在技术上是可行的，但到目前为止，缺乏分析技术阻碍了单个胚胎细胞中蛋白质、多肽和代谢物的表征。该项目将通过将细胞和发育生物学和神经科学的传统工具与生物分析化学的下一代仪器相结合，提供一种这样的技术创新--单细胞质谱仪。具体地说，光学显微镜、显微注射、微毛细管电泳法和纳米电喷雾电离将被应用于高分辨率串联质谱仪，以非洲爪哇和斑马鱼为模型，确定在胚胎的多个细胞中生物分子、蛋白质和代谢物的产生。该仪器将与领先的质谱学行业、生物学家和学生合作开发，并将由生物学家和使用这些重要发展模型的学生进行测试。

An award is made to the George Washington University to develop a device that will be able to measure the production of broad types of biomolecules in multiple, individual cells of the developing embryo. This project will enhance education by integrating biology and chemistry, and provide new investigative tools to raise creative research opportunities in basic and applied research. Development of the single-cell mass spectrometer will require regular interactions between analytical chemists, biologists, mass spectrometrists, and curators of data repositories, essentially creating an interdisciplinary environment for students and researchers to accomplish training beyond the classical curriculum in these disciplines. By demonstrating the device at the George Washington University and discussing its design, performance, and use at national conferences and publications, this work will broaden scientific literacy and inform of the availability of the device to a broader base of users. Data resulting from measurements on the production of biomolecules during embryogenesis will be disseminated in publicly accessible data repositories, providing a larger number of users with access to facilitate research and education in cell and developmental biology and neuroscience. Notably, the combination of these scientific and outreach elements, including participation of local high-school students in the project, will enhance research and education at the interface of biology, instrument development, and analytical chemistry, providing interdisciplinary solutions to current and future challenges in science and education. Characterization of biomolecular expression in single cells of the embryo will provide new insights into basic biochemical mechanisms that orchestrate embryonic development, the complex suite of processes by which a fertilized egg gives rise to an entire, fully functioning organism such as the frog, fish, or human. Although it has been technologically feasible to measure genes and transcripts in single embryonic cells, a lack of analytical technology has so far hindered the characterization of proteins, peptides, and metabolites in single embryonic cells. This project will provide one such technological innovation, a single-cell mass spectrometer, by combining traditional tools in cell and developmental biology and neuroscience with next-generation instrumentation from bioanalytical chemistry. Specifically, optical microscopy, microinjection, microcapillary electrophoresis, and nanoelectrospray ionization will be adapted to high-resolution tandem mass spectrometry to determine the production of biomolecules, proteins to metabolites, in multiple cells of the embryo using the frog Xenopus laevis and zebrafish as models. The instrument will be developed in collaboration with leading mass spectrometric industry, biologists, and students, and will be tested by biologists and students working with these important developmental models.