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首页> 外文期刊>Acta crystallographica. Section D, Biological crystallography. >A high-transparency, micro-patternable chip for X-ray diffraction analysis of microcrystals under native growth conditions
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A high-transparency, micro-patternable chip for X-ray diffraction analysis of microcrystals under native growth conditions

机译：透明度高,micro-patternable芯片x射线衍射分析微晶核本地生长条件

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摘要

Microcrystals present a significant impediment to the determination of macromolecular structures by X-ray diffraction methods. Although microfocus synchrotron beamlines and X-ray free-electron lasers (XFELs) can enable the collection of interpretable diffraction data from microcrystals, there is a need for efficient methods of harvesting small volumes (<2 mu l) of microcrystals grown under common laboratory formats and delivering them to an X-ray beam source under native growth conditions. One approach that shows promise in overcoming the challenges intrinsic to microcrystal analysis is to pair so-called 'fixed-target' sample-delivery devices with microbeam-based X-ray diffraction methods. However, to record weak diffraction patterns it is necessary to fabricate devices from X-ray-transparent materials that minimize background scattering. Presented here is the design of a new micro-diffraction device consisting of three layers fabricated from silicon nitride, photoresist and polyimide film. The chip features low X-ray scattering and X-ray absorption properties, and uses a customizable blend of hydrophobic and hydrophilic surface patterns to help localize microcrystals to defined regions. Microcrystals in their native growth conditions can be loaded into the chips with a standard pipette, allowing data collection at room temperature. Diffraction data collected from hen egg-white lysozyme microcrystals (10-15 mm) loaded into the chips yielded a complete, high-resolution (<1.6 angstrom) data set sufficient to determine a high-quality structure by molecular replacement. The features of the chip allow the rapid and user-friendly analysis of microcrystals grown under virtually any laboratory format at microfocus synchrotron beamlines and XFELs.
机译：微晶核形成重大障碍大分子结构的决心x射线衍射的方法。同步加速器beamlines和x射线自由电子激光(XFELs)可以启用的集合可说明的衍射数据微晶核,需要高效方法收集小卷(< 2μl)微晶核生长在普通实验室格式和送他们一束x射线源在本地生长条件。方法在克服显示承诺挑战固有的微晶分析对所谓的固定靶样品指定设备与microbeam-based x射线衍射方法。模式有必要制造设备从X-ray-transparent材料最小化背景散射。设计一种新的显微衍射装置组成的三层的氮化硅,光刻胶和聚酰亚胺薄膜。芯片功能低散射x射线和x射线吸收特性,并使用一个可定制的混合的疏水性和亲水性表面模式来帮助定位微晶核定义的区域。生长条件可以加载到芯片与一个标准的吸管,允许数据收集在室温下。从母鸡蛋清溶菌酶微晶核(10 - 15毫米)加载到芯片产生一个完整的、高分辨率(< 1.6埃)数据集足以确定一个高质量的结构通过分子替换。芯片允许快速和用户友好的分析在几乎任何微晶核的生长在微焦点同步加速器实验室格式beamlines XFELs。

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