Feasibility: Non-Contact Cryo-Atomic Force Microscope High Resolution Bioimaging

可行性：非接触式冷冻原子力显微镜高分辨率生物成像

• 批准号: 7024182
• 负责人: ZHIFENG SHAO
• 金额: $ 18.94万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7024182
• 关键词: Adhesions; Atomic Force Microscopy; Biological; Biological Preservation; Characteristics; Cleaved cell; Complex; Condition; Depth; Detection; Development; Dimensions; Electron Microscopy; Ensure; Environment; F-Actin; Feedback; Freezing; Frequencies; Goals; Helix (Snails); Heterogeneity; Image; In Situ; Individual; Integral Membrane Protein; Length; Lipid Bilayers; Liquid substance; Measures; Mechanics; Membrane; Methodology; Methods; Mica; Molecular; Nanotechnology; Nature; Nitrogen; Noise; Operative Surgical Procedures; Pliability; Rate; Resolution; Scanning; Scanning Probe Microscopes; Solutions; Specimen; Structure; Surface; Suspension substance; Suspensions; System; Temperature; Testing; Vacuum; base; bioimaging; cantilever; cold temperature; concept; cryogenics; design; ear helix; improved; instrumentation; macromolecule; nano; nanoscale; nanoscience; prevent; purge; ultra high resolution; vapor

DESCRIPTION (provided by applicant): One of the major objectives of the nanoscience and nanotechnology initiative is to develop general methodologies that can "effectively detect and analyze nanoscale entities of relevance to biomedicine". At present, supra-molecular complexes, structures with intrinsic flexibilities and/or heterogeneities and integral membrane proteins in the biological membrane remain a significant technological challenge. As such, the task of extracting "quantitative information (e.g., structural details) from biological nanoscale materials and machines" requires the development of more appropriate methods than those that must rely on ensemble averaging to achieve a high enough resolution. Here, we propose to develop an atomic force microscope (AFM) at cryogenic .temperatures to achieve a spatial resolution of 1 nm or better on individual complexes and nanoscale biological machines. The essential idea is to combine the high sensitivity of non-contact imaging and cryo-preservation of the native structure. Only with such an approach can the sharpness of an AFM tip be protected from degradation during imaging, a factor most critical for ultra-high resolution imaging. The immediate goal of this R21 application is to experimentally determine the feasibility of this approach and measure the fundamental parameters necessary for the design of robust instrumentation. If we succeed in developing this concept into a practical apparatus, such instrumentation will be a powerful method capable of resolving minute surface structural details (e.g., secondary structures) of individual complexes that cannot be surpassed by any other currently available approach.

描述（由申请人提供）：纳米科学和纳米技术计划的主要目标之一是开发可以“有效检测和分析与生物医学相关的纳米级实体”的通用方法。目前，生物膜中具有内在灵活性和/或异质性和积分膜蛋白的结构是一项重大的技术挑战。因此，与必须依靠合奏平均以实现足够高的分辨率的合奏相比，从生物纳米级材料和机器中提取“定量信息（例如结构细节）”的任务需要开发更合适的方法。在这里，我们提议在低温感应中开发原子力显微镜（AFM），以在单个复合物和纳米级生物机器上实现1 nm或更高的空间分辨率。重要的想法是结合非接触成像的高灵敏度和对天然结构的冷冻保护。只有采用这种方法，才能保护AFM尖端在成像过程中免受降解，这对于超高分辨率成像最重要的因素。该R21应用的直接目标是实验确定这种方法的可行性，并测量设计鲁棒仪器所需的基本参数。如果我们成功地将此概念发展为实用的设备，那么这种仪器将是一种强大的方法，能够解决单个复合体的微小表面结构细节（例如二级结构），这些细节无法超过任何其他当前可用的方法。