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Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers
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Chemistry
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JoVE Journal Chemistry
Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers

Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers

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8,871 Views

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07:26 min

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February 28, 2019

DOI:

10.3791/59087-v

DOI:
10.3791/59087-v

•

07:26 min
February 28, 2019

•

2 Views
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1Division of Biology and Chemistry – Laboratory for Biomolecular Research,Paul Scherrer Institut, 2Photon Science Division – SwissFEL,Paul Scherrer Institut, 3RIKEN SPring-8 Center, 4Department of Cell Biology, Graduate School of Medicine,Kyoto University, 5Department of Biology,ETH Zürich

Summary

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The success of a time-resolved serial femtosecond crystallography experiment is dependent on efficient sample delivery. Here, we describe protocols to optimize the extrusion of bacteriorhodopsin microcrystals from a high viscosity micro-extrusion injector. The methodology relies on sample homogenization with a novel three-way coupler and visualization with a high-speed camera.

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James, D., Weinert, T., Skopintsev, P., Furrer, A., Gashi, D., Tanaka, T., Nango, E., Nogly, P., Standfuss, J. Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers. J. Vis. Exp. (144), e59087, doi:10.3791/59087 (2019).

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