High Speed Atomic Force Microscopy for Real Time Imaging of Biological Processes

用于生物过程实时成像的高速原子力显微镜

• 批准号: 1063279
• 负责人: Todd Sulchek
• 金额: $ 34.31万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-02-15 至 2015-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1063279&HistoricalAwards=false
• 关键词: Speed; Atomic; Force; Microscopy; Real

AbstractIn all of its forms, the microscope is the most widely used tool in the investigation of biological structure and function. The study of living and moving biological systems, with sub-second time scale resolution and with nanometer length scale resolution, is becoming increasingly important in biological research. A critical example of biological research that requires faster, high resolution imaging of dynamic processes is the degradation of cellulosic materials by enzymatic, chemical, and physical processes. Improved cellulose degradation is an important step to efficient ethanol production and will likely be critical to the nation for development of a sustainable supply of fuel suitable for transportation. This project is to develop a system, based on the Atomic Force Microscope (AFM), for nanometer scale imaging of biological samples that is several orders of magnitude faster than current AFMs. The project will develop both new methodologies and instrumentation for the real-time and high-resolution imaging of dynamic biological processes. The project addresses a critical limitation for the AFM research community, while building upon the advances of current high-speed AFM systems. The technological advancement of this fast-scanning AFM is an active, self-actuating microcantilever. The system will also utilize a novel passivation scheme to provide robust operation in all liquid environments encountered in biological imaging. Moreover, the dimensions of the active probe will be miniaturized, to retain fast dynamics as well as delicate operation. The application of this fast-scanning system will be the real-time imaging of the degradation of cellulose by the cellulase enzyme from the fungus Trichonderma reesei. This system will visualize the mechanism of degradation and determine the kinetics of this process. The broader impact of this research will be to radically improve the characterization methods available to scientists and engineers working in biology and biotechnology. High speed scanning systems operating in microscopy labs across the country, where biologists could utilize video rate atomic force microscopy to obtain the fastest and highest resolution images of dynamic biological processes, would be enabled by this project. The microscope will also be capable of high speed molecular recognition for surface characterization of lipid membranes, proteins and molecular motors, cells and other biological systems. The initial step will be an investigation of the kinetic interactions between cellulose and cellulases, with the aim of improving production of cellulosic ethanol. An overarching priority for these advancements is to produce technology that is transferable to other commercial AFM systems, directly enabling the scanning probe research conducted at various research and industry labs. With the contributions of this project, dynamic, nanoscale imaging and manipulation can be extended to video rates, in a manner akin to scanning electron microscopy or video optical microscopy. The proposed program will also create educational opportunities for students to learn about the interface between physical science, biological science, and nanotechnology by focusing on graduate training, undergraduate involvement, and K-12 outreach efforts.

在各种形式中，显微镜是研究生物结构和功能最广泛使用的工具。对生命和运动的生物系统的研究，具有亚秒级的时间尺度分辨率和纳米级的长度尺度分辨率，在生物学研究中变得越来越重要。生物学研究的一个重要例子是需要更快、高分辨率的动态过程成像，这就是通过酶、化学和物理过程降解纤维素材料。改善纤维素降解是高效乙醇生产的重要一步，对于开发适合运输的可持续燃料供应可能至关重要。该项目旨在开发一种基于原子力显微镜（AFM）的系统，用于生物样品的纳米级成像，比目前的AFM快几个数量级。该项目将为动态生物过程的实时和高分辨率成像开发新的方法和仪器。该项目解决了AFM研究界的一个关键限制，同时建立在当前高速AFM系统的进步基础上。这种快速扫描原子力显微镜的技术进步是一个主动的，自驱动的微悬臂梁。该系统还将利用一种新的钝化方案，在生物成像中遇到的所有液体环境中提供稳健的操作。此外，有源探头的尺寸将被小型化，以保持快速的动态以及微妙的操作。该快速扫描系统的应用将是纤维素酶降解纤维素的实时成像从真菌里氏木霉。该系统将可视化降解机制并确定该过程的动力学。这项研究的更广泛影响将是从根本上改善生物学和生物技术领域的科学家和工程师可用的表征方法。在全国各地的显微镜实验室中运行的高速扫描系统，生物学家可以利用视频速率原子力显微镜获得动态生物过程的最快和最高分辨率的图像，将通过该项目实现。该显微镜还将能够高速分子识别，用于脂质膜，蛋白质和分子马达，细胞和其他生物系统的表面表征。第一步是研究纤维素和纤维素酶之间的动力学相互作用，目的是提高纤维素乙醇的生产。这些进步的首要任务是生产可转移到其他商业AFM系统的技术，直接实现在各种研究和工业实验室进行的扫描探针研究。随着该项目的贡献，动态，纳米级成像和操作可以扩展到视频速率，以类似于扫描电子显微镜或视频光学显微镜的方式。该计划还将为学生创造教育机会，通过专注于研究生培训，本科生参与和K-12外展工作，了解物理科学，生物科学和纳米技术之间的接口。