Current advances in synchrotron radiation instrumentation for MX experiments

Abstract

Following pioneering work 40 years ago, synchrotron beamlines dedicated to macromolecular crystallography (MX) have improved in almost every aspect as instrumentation has evolved. Beam sizes and crystal dimensions are now on the single micron scale while data can be collected from proteins with molecular weights over 10 MDa and from crystals with unit cell dimensions over 1000 Å. Furthermore it is possible to collect a complete data set in seconds, and obtain the resulting structure in minutes. The impact of MX synchrotron beamlines and their evolution is reflected in their scientific output, and MX is now the method of choice for a variety of aims from ligand binding to structure determination of membrane proteins, viruses and ribosomes, resulting in a much deeper understanding of the machinery of life. A main driving force of beamline evolution have been advances in almost every aspect of the instrumentation comprising a synchrotron beamline. In this review we aim to provide an overview of the current status of instrumentation at modern MX experiments. The most critical optical components are discussed, as are aspects of endstation design, sample delivery, visualization and positioning, the sample environment, beam shaping, detectors and data acquisition and processing.

Application of Synchrotron Radiation‐based Methods for Environmental Biogeochemistry: Introduction to the Special Section

“…Today, there are ∼50 synchrotron light sources worldwide and their beamline capabilities continue to evolve (Lightsources, 2016). This evolution will facilitate scientific discoveries (Owen et al, 2016) in many fields. A famous quote by Sydney Brenner in 1980, cited by many times before (Robertson, 1980), states that “progress in science depends on new techniques, new discoveries, and new ideas, probably in that order.” It is certainly well understood and accepted by environmental biogeochemists that there are significant challenges and limitations for any technique when diluted analytes are investigated in complex matrices, and that ongoing improvements in synchrotron techniques achieved through the availability of brighter light sources, better monochromators, and more sensitive detectors will help unlock new research possibilities and discoveries.…”

“…Today, there are ~50 synchrotron light sources worldwide and their beamline capabilities continue to evolve (Lightsources, 2016). This evolution will facilitate scientific discoveries (Owen et al, 2016) in many fields. A famous quote by Sydney Brenner in 1980, cited by many times before (Robertson, 1980), states that "progress in science depends on new techniques, new discoveries, and new ideas, probably in that order."…”

KAMO: towards automated data processing for microcrystals

“…The major difficulty in solving protein structures by X-ray crystallography is obtaining crystals with suitable size and sufficient diffracting power. Recently, X-ray microbeams have become available at synchrotron light sources and have enabled data collection from microcrystals at high signal-tonoise ratios (Smith et al, 2012;Owen et al, 2016;Yamamoto et al, 2017). The preparation of microcrystals is often easier than that of larger crystals, and smaller crystals may sometimes be better ordered and thus lead to higher quality data sets (Cusack et al, 1998;Evans et al, 2011).…”

A simple and versatile microfluidic device for efficient biomacromolecule crystallization and structural analysis by serial crystallography

“…Crystallography plays a central role in contemporary biology because it enables the visualization of the 3D architecture of biological macromolecules, which provides insights into their cellular functions and partnerships on the atomic scale (Giegé & Sauter, 2010;Jaskolski et al, 2014). Over the past two decades, the advent of structural genomics and associated high-throughput (HTP) technologies (Vincentelli et al, 2003;Pusey et al, 2005), together with dramatic improvements in experimental setups and the computational environment at synchrotron facilities (Terwilliger et al, 2009;Owen et al, 2016), have revolutionized the field and led to a torrent of new crystal structures. This productivity boost is clear from the number of structures deposited in the Protein Data Bank (PDB), which recently exceeded 150 000 entries.…”