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Fixed-target protein serial microcrystallography with an x-ray free electron laser

机译:具有X射线自由电子激光的固定靶蛋白连续微晶学

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摘要

We present results from experiments at the Linac Coherent Light Source (LCLS) demonstrating that serial femtosecond crystallography (SFX) can be performed to high resolution (~2.5 Å) using protein microcrystals deposited on an ultra-thin silicon nitride membrane and embedded in a preservation medium at room temperature. Data can be acquired at a high acquisition rate using x-ray free electron laser sources to overcome radiation damage, while sample consumption is dramatically reduced compared to flowing jet methods. We achieved a peak data acquisition rate of 10 Hz with a hit rate of ~38%, indicating that a complete data set could be acquired in about one 12-hour LCLS shift using the setup described here, or in even less time using hardware optimized for fixed target SFX. This demonstration opens the door to ultra low sample consumption SFX using the technique of diffraction-before-destruction on proteins that exist in only small quantities and/or do not produce the copious quantities of microcrystals required for flowing jet methods.
机译:我们提供直线加速器相干光源(LCLS)上的实验结果,证明使用沉积在超薄氮化硅膜上并嵌入防腐剂中的蛋白质微晶,可以实现高分辨率(〜2.5Å)的串联飞秒晶体学(SFX)在室温下为中等。使用X射线自由电子激光源可以克服辐射损伤,以高采集速率采集数据,而与流动喷射方法相比,样品消耗量大大减少。我们达到了10 acquisitionHz的峰值数据采集率,命中率约为38%,这表明使用此处介绍的设置可以在大约12小时的LCLS班次中获取完整的数据集,或者使用经过硬件优化的时间甚至可以在更短的时间内用于固定目标SFX。该演示使用销毁前仅对少量存在的蛋白质和/或不产生流动喷射方法所需的大量微晶的蛋白质进行衍射前衍射的技术,为超低样品消耗的SFX打开了大门。

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