EAGER: Field-Deployed Microfluidic Trap Array for Discovery and Observation of Microbial Eukaryotes

EAGER：用于发现和观察微生物真核生物的现场部署微流控陷阱阵列

• 批准号: 1027125
• 负责人: Leslie Shor
• 金额: $ 16万
• 依托单位: University of Connecticut
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1027125&HistoricalAwards=false
• 关键词: EAGER; Field; Deployed; Microfluidic; Trap

Bacteria and protozoa are critical components of aquatic and terrestrial ecosystems, driving biogeochemical processes including carbon fixation, oxygen production, nutrient cycling, and break-down of anthropogenic contaminants. In many habitats, the bacterial community structure and net production are controlled in a top-down fashion by protozoan predation; in turn, predation of protozoa by larger animals mobilizes nutrients and contaminants to higher trophic levels. Despite their importance, relatively little is known about the diversity, biogeography, and ecosystem function of microbial eukaryotes including protozoa, due to limitations of both traditional culturing methods and genetic techniques. New instrumentation is needed to sample, study and understand a broader range of protozoa in natural systems.The objective of this research is to develop a microfluidic field sampling and analysis tool to study the biogeography and function of microbial eukaryotes in natural habitats. Microfluidic samplers will be fabricated with micron-scale physical features in polydimethylsiloxane (PDMS) and glass. Samplers will be selectively baited and placed in different natural environments in order to select for microorganisms based on morphology, behavior, habitat, and prey source, and will enable direct observation of entrapped live protozoa via light microscopy. Genetic analysis of collected eukaryotes and prokaryotes will help provide a unified framework for taxonomic, functional, and genomic information. Together with co-located micro-scale measurements of environmental variables such as pH, temperature, and nutrient concentrations, this work will enable researchers to place microbial eukaryotes within the context of their immediate physical and chemical microhabitats.This work will enable development and validation of a completely new microbial community analysis tool that will be of immediate and practical use to microbial ecologists, aqueous geochemists, engineers, and marine scientists. It may help to answer fundamental questions in biology including what is the relevant physical scale of community structure variations, and to what extent are protozoan species cosmopolitan vs. endemic in natural habitats? Also, a better understanding of microbial community structure and function will permit better predictions of biogeochemical feedbacks as a function of climate change; improved understand the impacts of microbial community structure and function on contaminant uptake and mobilization, and enhanced risk models for environmental reservoirs of microbial pathogens. Future work will integrate a comprehensive suite of sensing capabilities and will deploy devices in a broader range of natural and engineered microbial habitats. The results of this project will be disseminated through peer-reviewed journals and conference presentations that target the biological user community. Implementation of the technology will be enabled by web-posted demonstration videos that illustrate how samplers are designed, created, tested, deployed, and studied back in the lab. All materials will be available online at http://www.cmbe.engr.uconn.edu/facultyshor.html.

细菌和原生动物是水生和陆地生态系统的重要组成部分，驱动着生物地球化学过程，包括碳固定，氧气生产，营养循环和人为污染物的分解。在许多生境中，细菌群落结构和净产量由原生动物捕食以自上而下的方式控制;反过来，大型动物捕食原生动物将营养物质和污染物转移到更高的营养水平。尽管它们的重要性，相对知之甚少微生物真核生物，包括原生动物的多样性，地理学，和生态系统功能，由于传统的培养方法和遗传技术的限制。本研究的目的是开发一种微流控现场采样和分析工具，用于研究自然环境中微生物真核生物的分布和功能。微流控采样器将在聚二甲基硅氧烷（PDMS）和玻璃中制造微米级的物理特征。采样器将被选择性地放置在不同的自然环境中，以根据形态、行为、栖息地和猎物来源选择微生物，并通过光学显微镜直接观察捕获的活原生动物。收集的真核生物和原核生物的遗传分析将有助于提供一个统一的框架，分类，功能和基因组信息。这项工作将使研究人员能够将微生物真核生物置于其直接的物理和化学微生境中。这项工作将使开发和验证一种全新的微生物群落分析工具成为可能，这将对微生物生态学家，水地球化学家，工程师，海洋科学家。它可能有助于回答生物学中的基本问题，包括群落结构变化的相关物理尺度是什么，以及原生动物物种在自然栖息地中的世界性与地方性在多大程度上？此外，更好地了解微生物群落结构和功能将允许更好地预测生物地球化学反馈作为气候变化的一个功能;更好地了解微生物群落结构和功能对污染物吸收和动员的影响，并增强微生物病原体环境水库的风险模型。未来的工作将整合一套全面的传感能力，并将在更广泛的自然和工程微生物栖息地部署设备。这一项目的成果将通过同行评审的期刊和针对生物学用户群体的会议报告进行传播。该技术的实施将通过网络发布的演示视频来实现，这些视频说明了采样器是如何设计、创建、测试、部署和在实验室中进行研究的。所有材料均可在http://www.cmbe.engr.uconn.edu/facultyshor.html网站上查阅。