Microfluidic Mixing Injector Holder Enables Routine Structural Enzymology Measurements with Mix-and-Inject Serial Crystallography Using X-ray Free Electron Lasers
Microfluidic Mixing Injector Holder Enables Routine Structural Enzymology Measurements with Mix-and-Inject Serial Crystallography Using X-ray Free Electron Lasers
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DOI：
10.1021/acs.analchem.9b00311
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发表时间：
2019-06-04
影响因子：
7.4
通讯作者：
Pollack, Lois
中科院分区：
文献类型：
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作者：
Calvey, George D.;Katz, Andrea M.;Pollack, Lois
X 射线自由电子激光源的混合注射串行晶体学 (MISC) 新兴技术提供了有关生物分子功能的原子级详细结构信息。尽管取得了早期成功，MISC 目前仍受到用于引发反应并将样品推进 X 射线束进行测量的混合注射器的效率和稳健性的限制。在这里，我们提出了一种制造注射器系统的新方法，该方法可以使实验更简单、更快、更有效。现在，可以在 100 分钟内用原材料生产出混合注射器，而实验过程中只需花费 5 分钟，从而节省了宝贵的时间。该系统是模块化的，可以在出现意外实验困难（例如堵塞）时快速更换部件。注射器支架设计灵活，允许优化每个设备，以最大限度地增加每次实验期间测量的衍射图案的数量。该支架已在两个不同的 X 射线自由电子激光源的四个光束时间内成功使用。其稳健性和易用性是使 MISC 技术易于使用和常规化的重要一步。
The emerging technique of Mix-and-Inject Serial Crystallography (MISC) at X-ray free electron laser sources provides atomically detailed structural information about biomolecules as they function. Despite early successes, MISC is currently limited by the efficiency and robustness of the mixing injectors used to initiate the reaction and propel the sample into the X-ray beam for measurement. Here, we present a new method for fabricating the injector system that leads to simpler, faster, and more effective experiments. A mixing injector can now be produced from raw components in 100 min, only 5 min of which must be spent during the experiment, saving valuable time. The system is modular, enabling parts to be quickly exchanged in the event of unanticipated experimental difficulties, such as clogging. The injector holder is designed to be flexible, allowing each device to be optimized to maximize the number of diffraction patterns measured during each experiment. This holder has been used successfully during four beamtimes at two different X-ray free electron laser sources. Its robustness and ease of use is an important step toward making the MISC technique accessible and routine.