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限制高亮度生物溶液散射的辐射损伤:欧洲分子生物学实验室P12光束线PETRAIII的实践经验。

Limiting radiation damage for high-brilliance biological solution scattering: practical experience at the EMBL P12 beamline PETRAIII.

作者信息

Jeffries Cy M, Graewert Melissa A, Svergun Dmitri I, Blanchet Clément E

机构信息

European Molecular Biology Laboratory Hamburg Outstation, c/o Deutsches Elektronen-Synchrotron, Notkestrasse 85, Hamburg 22603, Germany.

出版信息

J Synchrotron Radiat. 2015 Mar;22(2):273-9. doi: 10.1107/S1600577515000375. Epub 2015 Feb 4.

Abstract

Radiation damage is the general curse of structural biologists who use synchrotron small-angle X-ray scattering (SAXS) to investigate biological macromolecules in solution. The EMBL-P12 biological SAXS beamline located at the PETRAIII storage ring (DESY, Hamburg, Germany) caters to an extensive user community who integrate SAXS into their diverse structural biology programs. The high brilliance of the beamline [5.1 × 10(12) photons s(-1), 10 keV, 500 (H) µm × 250 (V) µm beam size at the sample position], combined with automated sample handling and data acquisition protocols, enable the high-throughput structural characterization of macromolecules in solution. However, considering the often-significant resources users invest to prepare samples, it is crucial that simple and effective protocols are in place to limit the effects of radiation damage once it has been detected. Here various practical approaches are evaluated that users can implement to limit radiation damage at the P12 beamline to maximize the chances of collecting quality data from radiation sensitive samples.

摘要

辐射损伤是使用同步加速器小角X射线散射(SAXS)来研究溶液中生物大分子的结构生物学家面临的普遍难题。位于德国汉堡DESY的PETRAIII储存环上的EMBL-P12生物SAXS光束线,服务于广泛的用户群体,他们将SAXS纳入各自不同的结构生物学研究项目中。该光束线具有高亮度[5.1×10(12) 个光子·秒(-1),10 keV,样品位置处的光束尺寸为500(水平)µm×250(垂直)µm],再加上自动化的样品处理和数据采集协议,能够对溶液中的大分子进行高通量结构表征。然而,考虑到用户通常投入大量资源来制备样品,一旦检测到辐射损伤,制定简单有效的方案以限制其影响至关重要。本文评估了各种实用方法,用户可以采用这些方法在P12光束线上限制辐射损伤,从而最大程度地提高从辐射敏感样品中收集高质量数据的机会。

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