Paul Scherrer Institut

3 ARTICLES PUBLISHED IN JoVE

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Chemistry

Improving High Viscosity Extrusion of Microcrystals for Time-resolved Serial Femtosecond Crystallography at X-ray Lasers
Daniel James 1, Tobias Weinert 1, Petr Skopintsev 1, Antonia Furrer 1, Dardan Gashi 1,2, Tomoyuki Tanaka 3,4, Eriko Nango 3,4, Przemyslaw Nogly 1,5, Joerg Standfuss 1
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

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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Biochemistry

Optimizing the Growth of Endothiapepsin Crystals for Serial Crystallography Experiments
John H. Beale 1, May E. Marsh 1
1Swiss Light Source, Paul Scherrer Institut

The aim of this article is to give the viewer a solid understanding of how to transform their small-volume, vapor-diffusion protocol, for growing large, single protein crystals, into a large-volume batch micro-crystallization method for serial crystallography.

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Biochemistry

A Sample Preparation Pipeline for Microcrystals at the VMXm Beamline
Adam D. Crawshaw 1, Emma V. Beale 1,2, Anna J. Warren 1, Andrew Stallwood 1,3, Graham Duller 1, Jose Trincao 1, Gwyndaf Evans 1,4
1Diamond Light Source Ltd, Harwell Science and Innovation Campus, 2Paul Scherrer Institut, 3Central Laser Facility, Science and Technologies Facilities Council, Harwell Science and Innovation Campus, 4Rosalind Franklin Institute, Harwell Science and Innovation Campus

The signal-to-noise ratio of data is one of the most important considerations in performing X-ray diffraction measurements from microcrystals. The VMXm beamline provides a low-noise environment and microbeam for such experiments. Here, we describe sample preparation methods for mounting and cooling microcrystals for VMXm and other microfocus macromolecular crystallography beamlines.

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