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Improved radiation dose efficiency in solution SAXS using a sheath flow sample environment.


ABSTRACT: 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.

SUBMITTER: Kirby N 

PROVIDER: S-EPMC5137223 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature

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Improved radiation dose efficiency in solution SAXS using a sheath flow sample environment.

Kirby Nigel N   Cowieson Nathan N   Hawley Adrian M AM   Mudie Stephen T ST   McGillivray Duncan J DJ   Kusel Michael M   Samardzic-Boban Vesna V   Ryan Timothy M TM  

Acta crystallographica. Section D, Structural biology 20161129 Pt 12


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  ...[more]

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