CAREER: IDBR: Ultrasmooth Patterned Metals for Membrane Biology

职业：IDBR：用于膜生物学的超光滑图案金属

• 批准号: 1054191
• 负责人: Sang-Hyun Oh
• 金额: $ 59.71万
• 依托单位: University of Minnesota-Twin Cities
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1054191&HistoricalAwards=false
• 关键词: CAREER; IDBR; Ultrasmooth; Patterned; Metals

Cell membranes are a complex mosaic of different lipids, with cholesterol, sphingolipids and membrane proteins forming nanoscale domains called lipid rafts. Lipid rafts transiently compartmentalize membrane constituents, which in turn mediate cellular functions in space and time. In order to quantitatively investigate the formation, morphology and signal transduction mechanisms mediated by lipid rafts and proteins integrated therein, they must be probed in artificial nano-environments that closely resemble real cell membranes. Analytical instruments that integrate lipid membranes with engineered solid-state sensors that can better mimic natural cellular environments will therefore offer deeper insights into important in vivo reactions and processes.The proposed platform will integrate soft lipid matter with the native sensing capability of nanoporous metallic films. Namely, surface plasmon resonance (SPR) and electrochemical sensing will be used to probe the biological processes mediated by lipid rafts and membrane proteins. However, noble metals such as gold or silver required for SPR have typically not been amenable to large-area, low-cost patterning at nanometer-scale resolution. To overcome this challenge, template-stripping methods will be used for reproducible high-throughput fabrication of ultrasmooth nanoporous metallic films that will be used as a multi-functional platform combining nanofluidics, optical detection, spectroscopy, and electrochemical sensing. The development of a membrane biosensing platform capable of molecular binding and transport assays will provide new tools to probe the dynamic heterogeneity of cellular membranes and the fundamental life processes they enable. Overall, disseminating these instruments will enable biologists to control, image, and quantitatively analyze lipid membranes and their constituents, potentially down to the single-molecule level, using precisely engineered biomimetic nano-environments.For broader outreach, the PI will organize "Sit with a Scientist" sessions and build an interactive Activity Station at the Science Museum of Minnesota during the one week-long NanoDays event in April of each year. The PI's exhibition booth at the museum will feature the theme of "Macro- to Microfluidics", with on-site demos of microfluidic devices for K-12 students. Graduate and undergraduate students associated with the project will gain interdisciplinary training ranging from nanofabrication and optics to chemistry and cellular membranes. Women and underrepresented minorities will be given opportunities to experience nanofabrication through NSF REU programs. The PI's new graduate course on biological instrumentation will integrate his research and education through theoretical lectures and weekly hands-on lab sessions to teach the construction and operation of membrane biosensors. New instruments and technology will be disseminated to the research community through an already active biannual, two-day bioMEMs short course. The process of making smooth patterned gold films, integration with microfluidics, lipid membrane formation, followed by SPR and electrochemical sensing will be demonstrated, allowing other researchers to duplicate the chip fabrication and instrument buildup.

细胞膜是不同脂质的复杂镶嵌体，胆固醇、鞘脂和膜蛋白形成称为脂筏的纳米级结构域。脂筏短暂地分隔膜成分，进而介导空间和时间上的细胞功能。为了定量研究脂筏和其中整合的蛋白质介导的形成、形态和信号转导机制，必须在与真实细胞膜非常相似的人工纳米环境中对其进行探测。因此，将脂质膜与能够更好地模拟自然细胞环境的工程固态传感器集成的分析仪器将为重要的体内反应和过程提供更深入的见解。所提出的平台将把软脂质物质与纳米多孔金属薄膜的天然传感能力集成起来。即，表面等离子共振（SPR）和电化学传感将用于探测脂筏和膜蛋白介导的生物过程。然而，SPR 所需的金或银等贵金属通常不适合在纳米级分辨率下进行大面积、低成本的图案化。为了克服这一挑战，模板剥离方法将用于超光滑纳米多孔金属薄膜的可重复高通量制造，该薄膜将用作结合纳米流体、光学检测、光谱学和电化学传感的多功能平台。能够进行分子结合和转运分析的膜生物传感平台的开发将为探测细胞膜的动态异质性及其所支持的基本生命过程提供新的工具。总体而言，传播这些仪器将使生物学家能够使用精确设计的仿生纳米环境来控制、成像和定量分析脂质膜及其成分，可能深入到单分子水平。为了更广泛的推广，PI 将组织“与每年四月为期一周的 NanoDays 活动期间，我们都会在明尼苏达州科学博物馆举办“a Scientist”课程并建立一个互动活动站。 PI在博物馆的展位将以“宏观到微流控”为主题，为K-12学生现场演示微流控装置。与该项目相关的研究生和本科生将获得从纳米制造和光学到化学和细胞膜的跨学科培训。女性和代表性不足的少数群体将有机会通过 NSF REU 项目体验纳米制造。 PI的新生物仪器研究生课程将通过理论讲座和每周的实验室实践课程将他的研究和教育结合起来，教授膜生物传感器的构建和操作。新仪器和技术将通过已经活跃的每两年一次、为期两天的生物 MEM 短期课程向研究界传播。将演示制作光滑图案金膜、与微流体集成、脂质膜形成、随后进行 SPR 和电化学传感的过程，使其他研究人员能够复制芯片制造和仪器构建。