Imaging system for high-throughput protein crystallization experiments

描述(申请人提供)：通过X射线结晶学获得的生物分子的高分辨率结构为理解功能和机制提供了最重要的数据来源。然而，结晶过程是一个主要的瓶颈，因为需要评估数百或数千个条件才能找到对结构确定有用的晶体。我们建议获得一个结晶实验的自动化可视化系统，以扩大可以有效筛选的结晶试验的数量，提高成功率。评估结晶试验的另一个挑战是区分蛋白质晶体和结晶实验中不需要的副产物的晶体，如盐，一个常见的问题，洗涤剂，或经常使涉及膜蛋白的实验复杂化的类脂。为了解决这些困难，我们选择了一种包括紫外线光学系统的仪器，该系统可以通过蛋白质晶体的吸收特性来识别蛋白质晶体。紫外光光学装置还提高了仪器的检测极限，因此可以放心地识别非常小的晶体(&lt；10&lt；m)。哥伦比亚大学有一个庞大而充满活力的结构生物学社区，参与了许多具有重大生物医学意义的项目，许多成员在NIH资助的高通量结晶学项目中发挥了主导作用，这些项目将直接受益于该仪器。最后，使用该仪器获得的图像将为哥伦比亚大学目前正在进行的自动图像分析软件的开发提供一个大型数据集。该软件将经过改进，以实现结晶条件的快速自动优化。在进行了大量的尽职调查后，我们选择提议收购Formulatrix成像系统，配备双紫外线功能，以及两个可在多种温度下使用的存储保温箱。 与公共健康相关：通过X射线结晶学获得的生物分子的高分辨率结构为理解功能和机制提供了最重要的数据来源。然而，结晶过程是一个主要的瓶颈，因为需要评估数百或数千个条件才能找到对结构确定有用的晶体。为了提高结晶实验的成功率，我们建议建立一个结晶实验的自动化可视化系统。

DESCRIPTION (provided by applicant): High-resolution structures of biological molecules obtained through x-ray crystallography provide among the most important sources of data for understanding function and mechanism. However, the process of crystallization presents a major bottleneck since hundreds or thousands of conditions need to be evaluated to find crystals useful for structure determination. We propose to acquire a system for the automated visualization of crystallization experiments in order to expand the number of crystallization trials that can be effectively screened, and enhance the rate of success. Another challenge in the evaluation of crystallization trials arises from distinguishing protein crystals from crystals of unwanted by-products of the crystallization experiment, such as salts, a common problem, and detergents, or lipids that frequently complicate experiments involving membrane proteins. To address these difficulties, we have selected an instrument that includes a UV optical system, which can identify protein crystals through their absorptive properties. UV optics also enhances the detection limit of the instrument such that very small crystals (<10<m) can be identified with confidence. Columbia University has a large and vibrant structural biology community, involved in numerous projects of substantial biomedical importance, and many members play leading role in NIH-funded high-throughput crystallography projects that will directly benefit from this instrument. Finally, the images acquired with this instrument will provide a large dataset for the development of automated image analysis software currently underway at Columbia. This software will be refined to enable rapid automated optimization of crystallization conditions. After substantial due diligence, we have opted to propose the acquisition of a Formulatrix imaging system, equipped with dual UV capability, and two storage incubators enabling use at multiple temperatures. PUBLIC HEALTH RELEVANCE: High-resolution structures of biological molecules obtained through x-ray crystallography provide among the most important sources of data for understanding function and mechanism. However, the process of crystallization presents a major bottleneck since hundreds or thousands of conditions need to be evaluated to find crystals useful for structure determination. We propose to acquire a system for the automated visualization of crystallization experiments in order to enhance the rate of success.