New techniques for detecting and handling nanocrystals for cutting edge structural biology methods

项目摘要 确定与人类健康和疾病相关的生物分子的详细结构特征 是我们的武器库中最重要的工具之一，用于了解疾病的病因和机制，以及 能够开发针对这些分子实体的新疗法。有一些新的技术在 结构生物学，包括系列飞秒结晶学、系列同步加速器结晶学，以及 微晶电子衍射，这有可能极大地提高结构测定 生物分子，并允许访问结构细节，这些细节已经通过其他fi描述 结构方法。这些新的结构化方法都依赖于能够生成、检测和 适当地处理极小的生物分子结晶样品。这项分项要求- 微米大小的晶体是这些技术的关键特征之一，也是一个主要障碍。 这些结构测定方法的进步。这项建议具有创新性 图像分析和样本处理技术都是专门为解决fic++规范而设计的。 处理亚微米晶体的挑战。我们计划使用非线性光学显微镜方法 与点过程建模和小波图像分析方法的专门应用相结合 提供能够检测和表征亚微米样品所需的计算工具 对于通常用于样品生成和分析的明亮的fi电子显微镜工具是不可见的 以晶体为基础的结构生物学实验装置。此外，我们还将检查不同的fixx目标 减少样品处理的平台，最大限度地减少潜在的晶体损害，以及测试声学的使用 用于纳升体积样品转移的液滴喷射技术。这些创新将是一个强大的 除了结构生物学工具箱，用于利用当前基于尖端绕射的方法 可用于结构确定。这些技术开发将突破关键障碍， 这些尖端结构方法的广泛使用。

Project Summary Determining the detailed structural characteristics of biomolecules relevant to human health and disease is one of the most crucial tools in our arsenal for understanding disease etiology and mechanism, and for being able to develop new therapeutics that target these molecular entities. There are new techniques in structural biology, including serial femtosecond crystallography, serial synchrotron crystallography, and microcrystal electron diffraction, that have the potential to greatly advance structure determination of biomolecules and to empower access to structural details that have defied characterization via other structural methods. These new structural methods all rely on being able to generate, detect and appropriately handle extremely small crystalline samples of biomolecules. This requirement for sub- micron sized crystals is one of the key features of these technologies, and presents a major obstacle to the advancement of these methods for structure determination. This proposal presents innovative technologies for both image analysis and sample handling expressly designed to address the specific challenges of working with submicron crystals. We plan to use nonlinear optical microscopy methods coupled with purpose-built application of point process modeling and wavelet image analysis approaches to provide computational tools needed to enable detection and characterization of submicron samples that are invisible to the brightfield microscopy tools that are typically used in sample generation and experimental set up for crystal based structural biology. In addition, we will examine different fixed target platforms to reduce sample handling, minimizing potential crystal damage, as well as test use of acoustic droplet ejection techniques for nanoliter volume sample transfer. These innovations will be a powerful addition to structural biology toolbox for leveraging the cutting edge diffraction based methods currently available for structure determination. These technology developments will break through key barriers to the widespread use of these cutting edge structural methods.