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

用于尖端结构生物学方法的检测和处理纳米晶体的新技术

• 批准号: 10705571
• 负责人: Sarah Elizabeth Johnson Bowman
• 金额: $ 38.8万
• 依托单位: HAUPTMAN-WOODWARD MEDICAL RESEARCH INST
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-17 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705571
• 关键词: Acceleration; Acoustics; Address; Algorithms; Binding; Biological; Biological Process; Brightfield Microscopy; Characteristics; Classification; Communities; Coupled; Cryoelectron Microscopy; Crystal Formation; Crystallization; Crystallography; Data; Deposition; Detection; Development; Diffusion; Disease; Drug Design; Etiology; Generations; Growth; Health; Human; Image; Image Analysis; Knowledge; Laboratories; Life; Liquid substance; Methods; Microscopy; Modeling; Molecular; Molecular Structure; Multimodal Imaging; Optics; Process; Property; Proteins; Research; Research Personnel; Resolution; Sampling; Solvents; Source; Structural Biologist; Structural Models; Structure; Synchrotrons; System; Techniques; Technology; Testing; Therapeutic; Time; Visible Radiation; Visualization; X-Ray Crystallography; X-Ray Medical Imaging; computerized tools; density; design; detection method; electron diffraction; empowerment; enzyme mechanism; experimental study; frontier; image registration; imaging modality; improved; innovation; innovative technologies; macromolecule; mathematical methods; method development; molecular targeted therapies; multimodality; nanocrystal; nanolitre; new therapeutic target; novel; optical imaging; particle; physical property; protein data bank; screening; skills; structural biology; submicron; technology development; three dimensional structure; tool

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.

项目概要 确定与人类健康和疾病相关的生物分子的详细结构特征 是我们了解疾病病因和机制的最重要工具之一， 能够开发针对这些分子实体的新疗法。其中有新的技术 结构生物学，包括串行飞秒晶体学、串行同步加速器晶体学和 微晶电子衍射，有可能极大地推进结构测定 生物分子，并能够获取通过其他方法无法表征的结构细节 结构方法。这些新的结构方法都依赖于能够生成、检测和 适当处理极小的生物分子晶体样品。此要求对于子 微米尺寸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。该提案提出了创新 专门为解决特定问题而设计的图像分析和样品处理技术 使用亚微米晶体的挑战。我们计划使用非线性光学显微镜方法 结合点过程建模和小波图像分析方法的专门应用 提供亚微米样品检测和表征所需的计算工具 对于通常用于样本生成和分析的明场显微镜工具来说是不可见的 基于晶体的结构生物学实验装置。此外，我们还将研究不同的固定目标 平台，以减少样品处理，最大限度地减少潜在的晶体损坏，以及测试使用声学 用于纳升体积样品转移的液滴喷射技术。这些创新将成为强大的 除了结构生物学工具箱之外，还可以利用当前最先进的基于衍射的方法 可用于结构测定。这些技术的发展将突破关键障碍 这些尖端结构方法的广泛使用。