Department of Chemical Engineering, The Pennsylvania State University, University Park, PA, United States.
Advanced Light Source and Materials Science Division, Lawrence Berkeley National Laboratory, Berkeley, CA, United States.
Methods Enzymol. 2022;677:357-383. doi: 10.1016/bs.mie.2022.08.042. Epub 2022 Nov 3.
The complex structure of biological assemblies is crucial for function yet challenging to discern given the chemical similarities between constituent components. Hard X-ray techniques, for example, rely on small density differences between domains that lead to modest scattering intensities. Resonant soft X-ray scattering (RSoXS) uses X-rays below 2keV to access absorption edges of low-Z elements. In this way, RSoXS can enhance scattering contrast between domains of different chemical compositions or bonding motifs, thus providing structural information about specific chemical motifs. RSoXS is emerging as a technique applicable for biological systems, having been used to characterize protein structure in solution and polysaccharide organization in plant cell walls. Sample environment instrumentation, however, is challenging in the current state of the art, particularly with liquid samples. This chapter contains a brief introduction to RSoXS and current beamline capabilities, and provides methods to prepare, store, and mount biological samples for RSoXS characterization. Furthermore, key details during RSoXS and X-ray absorption data acquisition are highlighted and some future opportunities in RSoXS instrumentation for biological systems are discussed.
生物组装体的复杂结构对于功能至关重要,但由于组成成分之间的化学相似性,难以辨别。例如,硬 X 射线技术依赖于导致适度散射强度的域之间的小密度差异。共振软 X 射线散射(RSoXS)使用低于 2keV 的 X 射线来获取低 Z 元素的吸收边。通过这种方式,RSoXS 可以增强不同化学成分或键合模式的域之间的散射对比度,从而提供关于特定化学基序的结构信息。RSoXS 作为一种适用于生物系统的技术正在兴起,已被用于表征溶液中的蛋白质结构和植物细胞壁中的多糖组织。然而,在当前的技术水平下,样品环境仪器设备具有挑战性,特别是对于液体样品。本章简要介绍了 RSoXS 和当前的光束线功能,并提供了用于 RSoXS 特征描述的准备、存储和安装生物样品的方法。此外,突出了在 RSoXS 和 X 射线吸收数据采集过程中的关键细节,并讨论了生物系统中 RSoXS 仪器设备的一些未来机会。