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

DOI:
10.3791/59087-v

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07:26 min

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February 28, 2019

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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

Chapters

  • 00:04Title
  • 00:25Protein Crystal Sample Preparation
  • 03:00Testing Sample Extrusion Using a High-Speed Camera Setup
  • 05:43Results: Extrusion of Protein Microcrystals from a High Viscosity Micro-extrusion Injector
  • 07:03Conclusion

Summary

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Automatic Translation

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.

Tags

Keywords: High Viscosity ExtrusionMicrocrystalsTime-resolved Serial Femtosecond CrystallographyX-ray LasersLCPMonooleinMAG 7.9Liquid ParaffinSyringeCubic PhaseExtrusion TestingHPLC Pump

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