SAXS/WAXS, Australian Synchrotron, 800 Blackburn Road, Clayton, Victoria 3168, Australia.
B21 SAXS, Diamond Light Source Ltd, Hartwell Science and Innovation Campus, Didcot OX11 0DE, England.
Acta Crystallogr D Struct Biol. 2016 Dec 1;72(Pt 12):1254-1266. doi: 10.1107/S2059798316017174. Epub 2016 Nov 29.
Radiation damage is a major limitation to synchrotron small-angle X-ray scattering analysis of biomacromolecules. Flowing the sample during exposure helps to reduce the problem, but its effectiveness in the laminar-flow regime is limited by slow flow velocity at the walls of sample cells. To overcome this limitation, the coflow method was developed, where the sample flows through the centre of its cell surrounded by a flow of matched buffer. The method permits an order-of-magnitude increase of X-ray incident flux before sample damage, improves measurement statistics and maintains low sample concentration limits. The method also efficiently handles sample volumes of a few microlitres, can increase sample throughput, is intrinsically resistant to capillary fouling by sample and is suited to static samples and size-exclusion chromatography applications. The method unlocks further potential of third-generation synchrotron beamlines to facilitate new and challenging applications in solution scattering.
辐射损伤是限制利用同步加速器小角 X 射线散射分析生物大分子的主要因素。在暴露过程中让样品流动有助于减少这个问题,但在层流状态下,其效果受到样品池壁处缓慢的流速限制。为了克服这个限制,发展出了共流法,其中样品在其被匹配缓冲液围绕的中心流过。该方法可以在样品损伤之前将 X 射线入射通量提高一个数量级,提高测量的统计数据,并保持低的样品浓度限制。该方法还能有效地处理几微升的样品体积,可以增加样品通量,本质上可以防止样品堵塞毛细作用,适用于静态样品和排阻色谱应用。该方法进一步挖掘第三代同步加速器光束线的潜力,以促进在溶液散射方面的新的和具有挑战性的应用。