European XFEL, Schenefeld, Germany.
European XFEL, Schenefeld, Germany.
Methods Enzymol. 2024;709:105-129. doi: 10.1016/bs.mie.2024.10.007. Epub 2024 Oct 22.
Serial femtosecond crystallography (SFX) at X-ray free electron lasers (XFELs) is a valuable technique for time-resolved structural studies on enzymes. This method allows for the collection of high-resolution datasets of protein structures at various time points during a reaction initiated by light or mixing. Experiments are performed under non-cryogenic conditions and allow the collection of radiation damage free structures. At the European XFEL (EuXFEL), SFX experiments are mainly performed with liquid jets produced by gas dynamic virtual nozzles (GDVNs) and less frequent with a high viscous extruder (HVE). In this chapter we describe these delivery methods, with the focus on GDVNs. Instrumentation, sample requirements, and preparation steps for SFX beamtimes are discussed. Other sample delivery methods available at the EuXFEL are briefly introduced at the end of this chapter.
连续飞秒晶体学(SFX)在 X 射线自由电子激光(XFEL)是一种有价值的技术,用于对酶进行时间分辨结构研究。该方法允许在光或混合引发的反应过程中,在不同时间点收集蛋白质结构的高分辨率数据集。实验在非低温条件下进行,并允许收集无辐射损伤的结构。在欧洲 X 射线自由电子激光(EuXFEL)上,SFX 实验主要使用气体动力学虚拟喷嘴(GDVN)产生的液体射流进行,而使用高粘性挤出机(HVE)的情况则较少。在本章中,我们将描述这些输送方法,重点介绍 GDVN。讨论了 SFX 光束时间的仪器、样品要求和制备步骤。本章结尾简要介绍了 EuXFEL 上提供的其他样品输送方法。