scholarly journals Versatile microporous polymer-based supports for serial macromolecular crystallography

2021 ◽  
Vol 77 (9) ◽  
pp. 1153-1167
Author(s):  
Isabelle Martiel ◽  
John H. Beale ◽  
Agnieszka Karpik ◽  
Chia-Ying Huang ◽  
Laura Vera ◽  
...  

Serial data collection has emerged as a major tool for data collection at state-of-the-art light sources, such as microfocus beamlines at synchrotrons and X-ray free-electron lasers. Challenging targets, characterized by small crystal sizes, weak diffraction and stringent dose limits, benefit most from these methods. Here, the use of a thin support made of a polymer-based membrane for performing serial data collection or screening experiments is demonstrated. It is shown that these supports are suitable for a wide range of protein crystals suspended in liquids. The supports have also proved to be applicable to challenging cases such as membrane proteins growing in the sponge phase. The sample-deposition method is simple and robust, as well as flexible and adaptable to a variety of cases. It results in an optimally thin specimen providing low background while maintaining minute amounts of mother liquor around the crystals. The 2 × 2 mm area enables the deposition of up to several microlitres of liquid. Imaging and visualization of the crystals are straightforward on the highly transparent membrane. Thanks to their affordable fabrication, these supports have the potential to become an attractive option for serial experiments at synchrotrons and free-electron lasers.

2019 ◽  
Vol 75 (2) ◽  
pp. 178-191 ◽  
Author(s):  
Marie Luise Grünbein ◽  
Gabriela Nass Kovacs

The high peak brilliance and femtosecond pulse duration of X-ray free-electron lasers (XFELs) provide new scientific opportunities for experiments in physics, chemistry and biology. In structural biology, one of the major applications is serial femtosecond crystallography. The intense XFEL pulse results in the destruction of any exposed microcrystal, making serial data collection mandatory. This requires a high-throughput serial approach to sample delivery. To this end, a number of such sample-delivery techniques have been developed, some of which have been ported to synchrotron sources, where they allow convenient low-dose data collection at room temperature. Here, the current sample-delivery techniques used at XFEL and synchrotron sources are reviewed, with an emphasis on liquid injection and high-viscosity extrusion, including their application for time-resolved experiments. The challenges associated with sample delivery at megahertz repetition-rate XFELs are also outlined.


2014 ◽  
Vol 369 (1647) ◽  
pp. 20130590 ◽  
Author(s):  
Jan Kern ◽  
Johan Hattne ◽  
Rosalie Tran ◽  
Roberto Alonso-Mori ◽  
Hartawan Laksmono ◽  
...  

X-ray free-electron lasers (XFELs) open up new possibilities for X-ray crystallographic and spectroscopic studies of radiation-sensitive biological samples under close to physiological conditions. To facilitate these new X-ray sources, tailored experimental methods and data-processing protocols have to be developed. The highly radiation-sensitive photosystem II (PSII) protein complex is a prime target for XFEL experiments aiming to study the mechanism of light-induced water oxidation taking place at a Mn cluster in this complex. We developed a set of tools for the study of PSII at XFELs, including a new liquid jet based on electrofocusing, an energy dispersive von Hamos X-ray emission spectrometer for the hard X-ray range and a high-throughput soft X-ray spectrometer based on a reflection zone plate. While our immediate focus is on PSII, the methods we describe here are applicable to a wide range of metalloenzymes. These experimental developments were complemented by a new software suite, cctbx.xfel . This software suite allows for near-real-time monitoring of the experimental parameters and detector signals and the detailed analysis of the diffraction and spectroscopy data collected by us at the Linac Coherent Light Source, taking into account the specific characteristics of data measured at an XFEL.


Author(s):  
Tetsuya Ishikawa

The evolution of synchrotron radiation (SR) sources and related sciences is discussed to explain the ‘generation’ of the SR sources. Most of the contemporary SR sources belong to the third generation, where the storage rings are optimized for the use of undulator radiation. The undulator development allowed to reduction of the electron energy of the storage ring necessary for delivering 10 keV X-rays from the initial 6–8 GeV to the current 3 Gev. Now is the transitional period from the double-bend-achromat lattice-based storage ring to the multi-bend-achromat lattice to achieve much smaller electron beam emittance. Free electron lasers are the other important accelerator-based light sources which recently reached hard X-ray regime by using self-amplified spontaneous emission scheme. Future accelerator-based X-ray sources should be continuous wave X-ray free electron lasers and pulsed X-ray free electron lasers. Some pathways to reach the future case are discussed. This article is part of the theme issue ‘Fifty years of synchrotron science: achievements and opportunities’.


2010 ◽  
Vol 03 (01) ◽  
pp. 93-120 ◽  
Author(s):  
Siegfried Schreiber

In the last couple of years, free electron lasers (FELs) have been a remarkable success as fourth generation light sources all over the world. Operating in the SASE mode, they produce laser-like radiation in a broad wavelength range. Especially in the soft and hard X-ray ranges, these light sources open unique and completely new fields in physics and allow a vast range of applications in most scientific fields. This article gives an overview of the principles of FELs and the SASE process, discusses technological challenges and solutions, and presents an outlook for future developments.


2010 ◽  
Vol 03 (01) ◽  
pp. 13-37 ◽  
Author(s):  
Jochen R. Schneider

Accelerator-based light sources stimulated progress in photon science in a truly extraordinary manner. The spectral brightness of storage-ring-based facilities increased by three orders of magnitude every 10 years since the 1960s. The extreme peak brightness at single-pass free electron X-ray lasers with pulse durations variable between about 1 and 300 femtoseconds will allow transformative experiments in many areas of science. This article is an attempt to show how progress in accelerator science and technology stimulated advancement in photon science, by discussing a limited number of examples of work at third generation storage ring facilities and free electron lasers. Hopes for further improvements in specific beam properties are expressed.


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