VMXi: a fully automated, fully remote, high-flux <i>in situ</i> macromolecular crystallography beamline

Sánchez-Weatherby, Juan; Sandy, James; Mikolajek, Halina; Lobley, C.M.C.; Mazzorana, Marco; Kelly, Jon; Preece, G.; Littlewood, Rich; Sørensen, Thomas

doi:10.1107/s1600577518015114

Cited by 40 publications

(32 citation statements)

References 34 publications

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“…Globally, the data can guide the development of future facilities by demonstrating that a range of beam diameters are required, even if larger complexes will be increasingly top-sliced by cryo-EM, and that the theoretical limits should be strived for in an experimental station. New, highly intense, beamlines with submicrometre beams are being constructed across the world that will be excellent for microcrystals and timeresolved experiments (Cohen et al, 2014;Sanchez-Weatherby et al, 2019). For more standard experiments, the data presented here demonstrate that microfocus beamlines will still be needed and, crucially, beamlines with larger focal spots should not be neglected.…”

Section: Resultsmentioning

confidence: 84%

A comparative anatomy of protein crystals: lessons from the automatic processing of 56 000 samples

Svensson

Gilski

Nurizzo

et al. 2019

IUCrJ

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The fully automatic processing of crystals of macromolecules has presented a unique opportunity to gather information on the samples that is not usually recorded. This has proved invaluable in improving sample-location, characterization and data-collection algorithms. After operating for four years, MASSIF-1 has now processed over 56 000 samples, gathering information at each stage, from the volume of the crystal to the unit-cell dimensions, the space group, the quality of the data collected and the reasoning behind the decisions made in data collection. This provides an unprecedented opportunity to analyse these data together, providing a detailed landscape of macromolecular crystals, intimate details of their contents and, importantly, how the two are related. The data show that mosaic spread is unrelated to the size or shape of crystals and demonstrate experimentally that diffraction intensities scale in proportion to crystal volume and molecular weight. It is also shown that crystal volume scales inversely with molecular weight. The results set the scene for the development of X-ray crystallography in a changing environment for structural biology.

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Section: Resultsmentioning

confidence: 84%

A comparative anatomy of protein crystals: lessons from the automatic processing of 56 000 samples

Svensson

Gilski

Nurizzo

et al. 2019

IUCrJ

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show abstract

“…In the developers' own words (Jacquamet et al, 2004) this new system completely changes the design of the experimental setup by merging functions that were previously considered to be distinct. This radical innovation was hailed as a 'seminal experiment' by the Diamond group (Aller et al, 2015) who have since built a whole beamline (VMX-i) dedicated to in situ crystallography (Sanchez-Weatherby et al, 2019). It also triggered the development of numerous new crystallization plates designed specifically to minimize the X-ray background and increase the rotation angle achievable in such in situ data collection experiments.…”

Section: Achievements and Legacymentioning

confidence: 99%

Jean-Luc Ferrer (1964–2020): structural biologist, beamline instrumentation innovator and entrepreneur

Bricogne¹

2020

Acta Cryst Sect D Struct Biol

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“…However, these delivery systems often suffer from a deficient hit rate and diminishingly minute yield, two metrics defined differently to measure the economy of crystal use (Ren et al, 2018), which severely limits their applicability to many biological systems. To address this challenge, a variety of fixed-target solutions have been proposed for serial crystallography (Baxter et al, 2016;Dhouib et al, 2009;Kisselman et al, 2011;le Maire et al, 2011;Perry et al, 2013;Wierman et al, 2019) and in situ diffraction (Axford et al, 2012;Bingel-Erlenmeyer et al, 2011;Sanchez-Weatherby et al, 2019), among which, we first proposed in situ serial Laue diffraction (Perry et al, 2014). However, implementation of an automated and robust platform for high-throughput serial data acquisition remains a challenging task for large-scale operation.…”

Section: Introductionmentioning

confidence: 99%

An automated platform for in situ serial crystallography at room temperature

Ren¹,

Shin²,

Bandara³

et al. 2020

IUCrJ

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Direct observation of functional motions in protein structures is highly desirable for understanding how these nanomachineries of life operate at the molecular level. Because cryogenic temperatures are non-physiological and may prohibit or even alter protein structural dynamics, it is necessary to develop robust X-ray diffraction methods that enable routine data collection at room temperature. We recently reported a crystal-on-crystal device to facilitate in situ diffraction of protein crystals at room temperature devoid of any sample manipulation. Here an automated serial crystallography platform based on this crystal-on-crystal technology is presented. A hardware and software prototype has been implemented, and protocols have been established that allow users to image, recognize and rank hundreds to thousands of protein crystals grown on a chip in optical scanning mode prior to serial introduction of these crystals to an X-ray beam in a programmable and high-throughput manner. This platform has been tested extensively using fragile protein crystals. We demonstrate that with affordable sample consumption, this in situ serial crystallography technology could give rise to room-temperature protein structures of higher resolution and superior map quality for those protein crystals that encounter difficulties during freezing. This serial data collection platform is compatible with both monochromatic oscillation and Laue methods for X-ray diffraction and presents a widely applicable approach for static and dynamic crystallographic studies at room temperature.

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VMXi: a fully automated, fully remote, high-flux in situ macromolecular crystallography beamline

Abstract: VMXi, a new tunable beamline dedicated to fully automatic screening and data collection from crystals in situ, is reported.

Cited by 40 publications

References 34 publications

A comparative anatomy of protein crystals: lessons from the automatic processing of 56 000 samples

A comparative anatomy of protein crystals: lessons from the automatic processing of 56 000 samples

Jean-Luc Ferrer (1964–2020): structural biologist, beamline instrumentation innovator and entrepreneur

An automated platform for in situ serial crystallography at room temperature

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