IIBR Instrumentation: Development of a Stimulated Raman Scattering Activated Cell Sorter to Enable Phenotype-Based Separation of Microbial Cells From Environmental Samples

授予蒙大拿州立大学开发受激拉曼散射激活细胞分选仪的奖项，以揭示环境微生物组中尚未表征的细胞的生物学。该仪器将由一个由三名研究生组成的团队建造和使用，他们在物理化学，机械工程和微生物学的交叉点工作。此外，6名本科生将在整个项目中接受培训。该项目的高度跨学科性质将保证为所有学生提供独特的培训体验。鉴于对微生物组及其对生态系统功能和人类健康的影响的浓厚兴趣，需要更好地了解微生物世界。该项目将培训中小学教育工作者，以帮助传播这一迅速发展的领域的标准和新发现。项目活动将针对博兹曼的萨卡加维亚中学的学生，每年的“蒙大拿州立大学家庭科学日”以及“科学技术峰会”，为期3天的年度研讨会，针对来自蒙大拿州的K-12教师。大多数微生物对实验室条件下培养的抗性阻碍了我们对微生物在其自然栖息地中的功能的理解。研究微生物的一种有效方法是在自然条件下以单细胞分辨率表征它们的生物学，并将它们的原位表型（它们在自然栖息地中的功能）和基因型（它们的遗传组成）联系起来。虽然有希望，但能够实现这种基于表型的细胞表征的可用仪器是众所周知的低通量，对细胞具有破坏性，或两者兼而有之。该项目将通过结合基于受激拉曼光谱的细胞表征、基于微流体的细胞分选和分选细胞的下游遗传表征来克服这些瓶颈。这种系统可实现的高细胞分选率将使整个微生物群落的基于表型的细胞分选成为可能，这是目前使用现有技术无法实现的。该项目将影响许多领域的研究人员，包括从事自然或人造系统研究的微生物生态学家，生物化学家，以及对研究单细胞生理学或稀有细胞类型生理学感兴趣的科学家。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估而被认为值得支持。

An award is made to Montana State University to develop a stimulated Raman scattering activated cell sorter to reveal the biology of yet uncharacterized cells in environmental microbiomes. The instrument will be built and utilized by a team of three graduate students working at the intersection of physical chemistry, mechanical engineering, and microbiology. In addition, six undergraduate students will be trained throughout the project. The highly interdisciplinary nature of the project will guarantee a unique training experience for all students. Given the intense interest associated with the microbiome and its effects on ecosystem functioning and human health, a better understanding of the microbial world is needed. This project will train primary and secondary educators to help communicate standards and new findings in this rapidly evolving field. Project activities will target students at Bozeman’s Sacagawea Middle School, the annual “Montana State University Family Science Day” as well as the “Science-Technology Summit”, a 3-day annual workshop targeted at K-12 teachers from across the state of Montana.The resistance to cultivation under laboratory conditions of most microorganisms has hindered our understanding of the functioning of microbes in their natural habitats. A powerful way to study microbes is to characterize their biology under natural conditions at single-cell resolution and link their in situ phenotype - their function in their natural habitat - and genotype - their genetic make-up. While promising, the available instrumentation capable of achieving such phenotype-based cell characterization is notoriously low throughout, destructive to the cell, or both. This project will overcome these bottlenecks by combining stimulated Raman spectroscopy-based cell characterization, microfluidics-based cell sorting, and downstream genetic characterization of sorted cells. The high cell sorting rates achievable by such a system will enable phenotype-based cell sorting of entire microbial communities, something that is currently not achievable using existing technologies. The project will impact researchers across many fields, including microbial ecologists working on natural or human-made system, biogeochemists, and scientists interested in studying single-cell physiology or the physiology of rare cell types.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.