System for generation of frozen-hydrated samples using the Focused Ion Beam

项目摘要 我们应用程序的长期目标是开发硬件和程序 这将使研究人员更好地了解细胞的功能。我们的 具体目标是提供一个系统，使研究人员能够更好地准备 冷冻水化标本通过断层重建进行研究 透射式电子显微镜 目前使用冷冻-水合样品制备冷冻水化样品的方法 超微切割术产生具有组织变形的波状样本，需要 投入大量的时间和耐心来获得即使是低收益的结果。一则广告 聚焦离子束(FIB)系统是一种很有前途的样品分析工具 准备工作，但最新研究表明，目前可用的系统 与FIB一起使用，允许样品污染(主要是霜冻)，并具有较差的 这项技术的样品处理硬件。 与生物可视化资源密切合作 沃兹沃斯中心NIH生物技术资源的复杂性(RVBC)， 我们将开发一种新的样品架，展示这一点的顺利转移 取样进出FIB，演示基本的FIB切割图案和 最终显示出优越的断层图像。我们的意图是发展和 将经济高效的系统商业化，使这些技术能够 为生物研究界提供广泛的信息。项目说明： 这项提议将开发工具，使生物学家能够更清楚地了解 细胞的结构和功能。这方面的知识是无数领域的基础。 医学研究和实践。

Project Summary The long term objective of our application is to develop hardware and procedures that will allow researchers to better understand the function of the cell. Our specific aim is to provide a system that will allow researches to better prepare frozen hydrated specimens for study via tomographic reconstruction in the transmission electron microscope(TEM) Current methods of preparing frozen hydrated specimens using cryo- ultramicrotomy produce wavy samples with tissue deformation and require a large investment of time and patience to get even low yield results. A commercial focused ion beam (FIB) system is a promising alternative tool for sample preparation, but the latest research suggests that currently available systems for use with the FIB allow sample contamination (primarily frost) and have poor sample handling hardware for this technique. In close collaboration with the Resource for the Visualization of Biological Complexity (RVBC) at the Wadsworth Center's NIH Biotechnological Resource, we will develop a new sample holder, demonstrate smooth transfer of this sample into and out of the FIB, demonstrate basic FIB cutting patterns and ultimately show superior tomograms. It is our intent to develop and commercialize a cost effective system that will enable these techniques to become widely available to the biology research community. Project Narrative: 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.