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.

向乔治华盛顿大学颁发了奖励，以开发一种能够衡量发育中胚胎多种单个细胞中广泛的生物分子的产生的设备。该项目将通过整合生物学和化学来增强教育，并提供新的调查工具来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，质谱师和数据存储库的策展人之间进行定期相互作用，从本质上为学生和研究人员创造了跨学科环境，以完成这些学科中经典课程以外的培训。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中讨论其设计，性能和使用，这项工作将扩大科学素养，并将设备的可用性告知更广泛的用户基础。通过公开访问的数据存储库来传播有关胚胎发生过程中生物分子生物分子的测量结果的数据，从而为更多的用户提供了访问的访问，可以促进细胞，发育生物学和神经科学方面的研究和教育。值得注意的是，这些科学和外展元素的结合，包括当地高中生参与该项目，将在生物学，仪器开发和分析化学的界面上增强研究和教育，从而为科学和教育方面的当前和未来挑战提供跨学科的解决方案。在胚胎的单个细胞中生物分子表达的表征将提供对编排胚胎发育的基本生物化学机制的新见解，这些机制是胚胎发育，这是一个复杂的过程套件，通过该机制，受精卵会产生整个功能齐全的生物体，例如青蛙，鱼类或人类。尽管在单个胚胎细胞中测量基因和转录本在技术上是可行的，但缺乏分析技术已经阻碍了单个胚胎细胞中蛋白质，肽和代谢产物的表征。该项目将通过将传统的细胞和发育生物学工具和神经科学与生物分析化学的下一代仪器相结合，从而提供一种这样的技术创新，即单细胞质谱仪。具体而言，光学显微镜，显微注射，显微毛细血管电泳和纳米电喷雾电离将适用于高分辨率的串联质谱法，以确定生物分子，蛋白质，蛋白质的蛋白质，用于代谢物，使用Frog Xenopus laevis laevis laevis and Zebrafish AS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，并将由使用这些重要发展模型的生物学家和学生进行测试。