Supplement for System for generation of frozen-hydrated samples using the Focused

使用 Focused 生成冷冻水合样品的系统的补充

• 批准号: 7671691
• 负责人: Norman James Salmon
• 金额: $ 35万
• 依托单位: HUMMINGBIRD SCIENTIFIC, LLC
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-07-15 至 2011-07-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7671691
• 关键词: 3-Dimensional; Area; Award; Biological; Biology; Cell physiology; Cells; Cellular Structures; Communities; Complex; Computer software; Coupling; Crowding; Custom; Data Collection; Development; Devices; Documentation; Electron Beam; Electron Microscope; Electron Microscopy; Freezing; Generations; Hour; Ice; Image; In Situ; Industry; Interruption; Ions; Knowledge; Laboratories; Lifting; Liquid substance; Location; Mainstreaming; Mammalian Cell; Manuals; Mechanics; Medical Research; Metals; Methods; Modeling; Modification; Morphologic artifacts; Nanotechnology; Nitrogen; Operative Surgical Procedures; Organelles; Phase; Positioning Attribute; Preparation; Proteins; Research; Resolution; Risk; Sampling; Scientist; Semiconductors; Side; Site; Small Business Innovation Research Grant; Specimen; Staging; Structure; Surface; System; Techniques; Temperature; Thick; Three-Dimensional Imaging; Time; Tissues; Tomogram; Ultramicrotomy; Universities; Work; Writing; base; biological research; cost; design; electron tomography; flexibility; improved; in vivo; instrument; interest; light microscopy; meetings; nanoscale; pressure; prototype; public health relevance; reconstruction; research study; tomography; tool; transmission process; vibration

DESCRIPTION (provided by applicant): Transmission electron microscope (TEM) tomography provides three-dimensional images at the nanometer scale. It is a mainstream technique in biological electron microscopy, and is rapidly finding applications in the semiconductor and nanotechnology industries. In biology, scientists are interested in producing faithful tomograms of frozen-hydrated samples, since specimens prepared in this way are considered to represent the native in vivo structure. In the case of typical mammalian cells and tissue, such specimens are usually high- pressure frozen, and then prepared for electron tomography using cryo-ultramicrotomy. However the difficulties of cryo-ultramicrotomy, and the artifacts that are unavoidably created by mechanical sectioning, have discouraged most laboratories from attempting study of frozen-hydrated cells and tissue by electron tomography. Preliminary work by our Wadsworth collaborators has shown that use of a cryo-FIB system avoids the artifacts and difficulties of cryo-ultramicrotomy, and offers additional advantages in preparing frozen-hydrated specimens. PUBLIC HEALTH RELEVANCE: This proposal will develop tools that allow biologists to get a clearer picture of the structure and function of the cell. This knowledge is fundamental to innumerable areas of medical research and practice.

描述（由申请人提供）：透射电子显微镜（TEM）断层扫描提供纳米级的三维图像。它是生物电子显微镜的主流技术，并迅速在半导体和纳米技术行业中找到应用。在生物学中，科学家们对制作冷冻水合样品的真实断层图像感兴趣，因为以这种方式制备的标本被认为代表了体内的天然结构。在典型的哺乳动物细胞和组织的情况下，这样的标本通常被高压冷冻，然后使用冷冻超微切片术制备用于电子断层扫描。然而，冷冻超微切片术的困难，以及机械切片可能产生的伪影，阻碍了大多数实验室尝试通过电子断层扫描研究冷冻水合细胞和组织。我们的沃兹沃斯合作者的初步工作表明，使用冷冻FIB系统避免了冷冻超微切片术的伪影和困难，并在制备冷冻水合标本方面提供了额外的优势。公共卫生相关性：该提案将开发工具，使生物学家能够更清楚地了解细胞的结构和功能。这些知识是医学研究和实践的无数领域的基础。