X-CHIP: an integrated platform for high-throughput protein crystallization and on-the-chip X-ray diffraction data collection.
X-CHIP: an integrated platform for high-throughput protein crystallization and on-the-chip X-ray diffraction data collection.
复制标题
DOI：
10.1107/s0907444911011589
复制
发表时间：
2011-06
期刊：
影响因子：
--
通讯作者：
Chirgadze NY
中科院分区：
文献类型：
--
作者：
Kisselman G;Qiu W;Romanov V;Thompson CM;Lam R;Battaile KP;Pai EF;Chirgadze NY
X-CHIP（X射线晶体学高通量集成平台）是一种新型微芯片，它已被开发用于将从结晶到衍射数据收集的晶体学管道的多个步骤结合在单个设备上，以简化整个过程。X-CHIP（X射线结晶高通量集成平台）是一种新型的微芯片，其开发目的是将从结晶到衍射数据收集的晶体学管道的多个步骤联合收割机结合在单个设备上，以简化整个过程。该系统设计用于结晶条件筛选、晶体目视检查、初始X射线筛选和高通量数据收集。X射线衍射数据采集可以使用原位方法在室温下直接在芯片上进行。该芯片的功能消除了手动晶体处理和晶体样品冷冻保护的必要性，同时允许从同一滴中的多个晶体收集数据。这项技术对于具有大量数据的项目特别有益，例如蛋白质复合物研究和基于片段的筛选。该平台在对可见光和X射线透明的微型板上采用亲水性和疏水性同心环表面，以创建明确且稳定的微批结晶环境。结晶实验和数据采集实验结果表明，该技术可以生长出高质量的衍射晶体，并可以采集高分辨率的衍射数据集。此外，在室温下用X-CHIP收集的单波长异常色散数据集的质量足以生成可解释的电子密度图。由于使用了纳升级的液滴体积，该技术具有很高的资源效率。它不需要对大多数内部和同步加速器光束线系统进行任何修改，并为X射线结构测定过程的完全自动化提供了一个有前途的机会。
The X-CHIP (X-ray Crystallography High-throughput Integrated Platform) is a novel microchip that has been developed to combine multiple steps of the crystallographic pipeline from crystallization to diffraction data collection on a single device to streamline the entire process. The X-CHIP (X-ray Crystallization High-throughput Integrated Platform) is a novel microchip that has been developed to combine multiple steps of the crystallographic pipeline from crystallization to diffraction data collection on a single device to streamline the entire process. The system has been designed for crystallization condition screening, visual crystal inspection, initial X-ray screening and data collection in a high-throughput fashion. X-ray diffraction data acquisition can be performed directly on-the-chip at room temperature using an in situ approach. The capabilities of the chip eliminate the necessity for manual crystal handling and cryoprotection of crystal samples, while allowing data collection from multiple crystals in the same drop. This technology would be especially beneficial for projects with large volumes of data, such as protein-complex studies and fragment-based screening. The platform employs hydrophilic and hydrophobic concentric ring surfaces on a miniature plate transparent to visible light and X-rays to create a well defined and stable microbatch crystallization environment. The results of crystallization and data-collection experiments demonstrate that high-quality well diffracting crystals can be grown and high-resolution diffraction data sets can be collected using this technology. Furthermore, the quality of a single-wavelength anomalous dispersion data set collected with the X-CHIP at room temperature was sufficient to generate interpretable electron-density maps. This technology is highly resource-efficient owing to the use of nanolitre-scale drop volumes. It does not require any modification for most in-house and synchrotron beamline systems and offers a promising opportunity for full automation of the X-ray structure-determination process.