European Synchrotron Radiation Facility, B.P. 220, F-38043 Grenoble Cedex, France.
Anal Chem. 2011 Jun 15;83(12):4863-70. doi: 10.1021/ac200515x. Epub 2011 May 18.
Investigations of single fragile objects manipulated by optical forces with high brilliance X-ray beams may initiate the development of new research fields such as protein crystallography in an aqueous environment. We have developed a dedicated optical tweezers setup with a compact, portable, and versatile geometry for the customary manipulation of objects for synchrotron radiation applications. Objects of a few micrometers up to a few tens of micrometers size can be trapped for extended periods of time. The selection and positioning of single objects out of a batch of many can be performed semi-automatically by software routines. The performance of the setup has been tested by wide-angle and small-angle X-ray scattering experiments on single optically trapped starch granules, using a synchrotron radiation microbeam. We demonstrate here for the first time the feasibility of microdiffraction on optically trapped protein crystals. Starch granules and insulin crystals were repeatedly raster-scanned at about 50 ms exposure/raster-point up to the complete loss of the structural order. Radiation damage in starch granules results in the appearance of low-angle scattering due to the breakdown of the polysaccharide matrix. For insulin crystals, order along the densely packed [110] direction is preferentially maintained until complete loss of long-range order.
用高亮度 X 射线束研究单个易碎物体的光学操纵可能会开创蛋白质晶体在水相环境中的新研究领域。我们开发了一种专用的光学镊子装置,其结构紧凑、便携且用途广泛,可用于在同步加速器辐射应用中对物体进行常规操纵。几个微米到几十微米大小的物体可以被长时间捕获。通过软件例程,可以半自动地从一批物体中选择和定位单个物体。该装置的性能已通过在同步加速器微束上对单个光学捕获的淀粉颗粒进行广角和小角 X 射线散射实验进行了测试。我们首次展示了在光学捕获的蛋白质晶体上进行微衍射的可行性。淀粉颗粒和胰岛素晶体以约 50 ms 的曝光/光栅点重复进行光栅扫描,直到结构有序完全丧失。淀粉颗粒中的辐射损伤导致低角度散射的出现,这是由于多糖基质的破坏。对于胰岛素晶体,沿密集排列的[110]方向的有序性优先保持,直到长程有序完全丧失。