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

项目摘要 我们申请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 具体目标是提供一个系统，使研究更好地准备 通过断层重建研究的冷冻水合标本， 透射电子显微镜 目前使用低温制备冷冻水合样品的方法， 超薄切片术产生具有组织变形的波浪状样品， 投入大量的时间和耐心，以获得即使是低产量的结果。商业 聚焦离子束（FIB）系统是一种很有前途的替代工具， 准备，但最新的研究表明，目前可用的系统， 与FIB一起使用允许样品污染（主要是霜）， 用于该技术的样品处理硬件。 在与生物可视化资源的密切合作下， 复杂性（RVBC）在沃兹沃斯中心的NIH生物技术资源， 我们将开发一种新的样品保持器，证明这种保持器的平稳转移。 从FIB中取样，演示基本的FIB切割模式， 最终显示出较好的上级断层图像。我们的目的是发展和 商业化的成本效益系统，使这些技术， 在生物学研究界广泛使用。项目叙述： 这项提案将开发工具，使生物学家能够更清楚地了解 细胞的结构和功能。这一知识对无数领域都至关重要 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.