用于欧洲 X 射线自由电子激光的串行晶体学的分段流发生器。
Segmented flow generator for serial crystallography at the European X-ray free electron laser.
机构信息
School of Molecular Sciences, Arizona State University, Tempe, AZ, 85287-1604, USA.
Center for Applied Structural Discovery, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287-7401, USA.
出版信息
Nat Commun. 2020 Sep 9;11(1):4511. doi: 10.1038/s41467-020-18156-7.
Serial femtosecond crystallography (SFX) with X-ray free electron lasers (XFELs) allows structure determination of membrane proteins and time-resolved crystallography. Common liquid sample delivery continuously jets the protein crystal suspension into the path of the XFEL, wasting a vast amount of sample due to the pulsed nature of all current XFEL sources. The European XFEL (EuXFEL) delivers femtosecond (fs) X-ray pulses in trains spaced 100 ms apart whereas pulses within trains are currently separated by 889 ns. Therefore, continuous sample delivery via fast jets wastes >99% of sample. Here, we introduce a microfluidic device delivering crystal laden droplets segmented with an immiscible oil reducing sample waste and demonstrate droplet injection at the EuXFEL compatible with high pressure liquid delivery of an SFX experiment. While achieving ~60% reduction in sample waste, we determine the structure of the enzyme 3-deoxy-D-manno-octulosonate-8-phosphate synthase from microcrystals delivered in droplets revealing distinct structural features not previously reported.
连续飞秒晶体学（SFX）与 X 射线自由电子激光（XFEL）结合使用，可以确定膜蛋白的结构并进行时间分辨晶体学研究。常见的液体样品输送方式是将蛋白质晶体悬浮液连续喷射到 XFEL 的路径中，但由于所有当前 XFEL 源的脉冲性质，会浪费大量的样品。欧洲 X 射线自由电子激光（EuXFEL）以 100 毫秒的间隔发射飞秒（fs）X 射线脉冲，而当前在脉冲之间的间隔为 889 纳秒。因此，通过快速喷射进行连续样品输送会浪费超过 99%的样品。在这里，我们引入了一种微流控装置，用于输送带有不混相油的晶体负载液滴，以减少样品浪费，并在与 SFX 实验的高压液体输送兼容的条件下，在 EuXFEL 上演示液滴注入。在实现约 60%的样品浪费减少的同时，我们确定了来自液滴中微晶体的 3-脱氧-D-甘露庚酮糖-8-磷酸合酶的结构，揭示了以前未报道的独特结构特征。
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