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

IDBR：A 型 - 开发用于绘制发育中胚胎中小分子表达图谱的原位单细胞质谱仪

• 批准号: 1455474
• 负责人: Peter Nemes
• 金额: $ 39.59万
• 依托单位: George Washington University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-05-01 至 2018-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1455474&HistoricalAwards=false
• 关键词: IDBR; TYPE; Development; situ; Single

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.

乔治华盛顿大学获得了一项奖励，以开发一种能够测量发育中胚胎的多个单个细胞中多种生物分子的产生的装置。该项目将通过整合生物学和化学来加强教育，并提供新的研究工具，以提高基础和应用研究中的创造性研究机会。单细胞质谱仪的开发需要分析化学家，生物学家，质谱学家和数据库管理员之间的定期互动，基本上为学生和研究人员创造了一个跨学科的环境，以完成这些学科的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其设计、性能和使用，这项工作将扩大科学素养，并向更广泛的用户群宣传该设备的可用性。胚胎发生过程中生物分子产生的测量数据将在可公开访问的数据库中传播，使更多用户能够访问，以促进细胞和发育生物学以及神经科学方面的研究和教育。值得注意的是，这些科学和推广元素的结合，包括当地高中学生参与该项目，将加强生物学，仪器开发和分析化学界面的研究和教育，为当前和未来的科学和教育挑战提供跨学科的解决方案。胚胎单细胞中生物分子表达的表征将为协调胚胎发育的基本生化机制提供新的见解，胚胎发育是一系列复杂的过程，受精卵通过这些过程产生一个完整的，功能齐全的生物体，如青蛙，鱼或人类。虽然在单个胚胎细胞中测量基因和转录本在技术上是可行的，但迄今为止，缺乏分析技术阻碍了单个胚胎细胞中蛋白质、肽和代谢物的表征。该项目将提供一种这样的技术创新，即单细胞质谱仪，将细胞和发育生物学和神经科学中的传统工具与生物分析化学的下一代仪器相结合。具体而言，光学显微镜，显微注射，微毛细管电泳和纳米电喷雾电离将适用于高分辨率串联质谱法，以确定生产的生物分子，蛋白质代谢产物，在多个细胞的胚胎使用青蛙非洲爪蟾和斑马鱼作为模型。该仪器将与领先的质谱行业，生物学家和学生合作开发，并将由生物学家和学生使用这些重要的发展模型进行测试。