Paul Scherrer Institut

3 ARTICLES PUBLISHED IN JoVE

Chemistry

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

Daniel James¹, Tobias Weinert¹, Petr Skopintsev¹, Antonia Furrer¹, Dardan Gashi^1,2, Tomoyuki Tanaka^3,4, Eriko Nango^3,4, Przemyslaw Nogly^1,5, Joerg Standfuss¹

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.

Biochemistry

Optimizing the Growth of Endothiapepsin Crystals for Serial Crystallography Experiments

John H. Beale¹, May E. Marsh¹

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.

Biochemistry

A Sample Preparation Pipeline for Microcrystals at the VMXm Beamline

Adam D. Crawshaw¹, Emma V. Beale^1,2, Anna J. Warren¹, Andrew Stallwood^1,3, Graham Duller¹, Jose Trincao¹, 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.