Characterization and use of the spent beam for serial operation of LCLS

Boutet, Sébastien; Foucar, L.; Barends, Thomas R. M.; Botha, Sabine; Doak, R. Bruce; Koglin, Jason E.; Messerschmidt, Marc; Nass, Karol; Schlichting, Ilme; Seibert, M.; Shoeman, Robert L.; Williams, Garth J.

doi:10.1107/s1600577515004002

Cited by 17 publications

(14 citation statements)

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“…The experiments were performed using a serial operation of the serial crystallography set-up at the Coherent X-ray Imaging instrument (CXI) of LCLS in a manner similar to a previous report21: the unused beam from an upstream experiment was refocused for a downstream experiment, allowing simultaneous data collection from two separate sample chambers. Individual X-ray pulses of nominally 12.8 keV photon energy and 0.93 mJ average pulse energy with pulse duration between 35 and 40 fs were focused into a spot size of approximately 3 × 3 μm in both the upstream and downstream interaction regions using beryllium compound refractive lenses.…”

Section: Resultsmentioning

confidence: 99%

“…Individual X-ray pulses of nominally 12.8 keV photon energy and 0.93 mJ average pulse energy with pulse duration between 35 and 40 fs were focused into a spot size of approximately 3 × 3 μm in both the upstream and downstream interaction regions using beryllium compound refractive lenses. Microcrystals of the Se-B SA complex were delivered to the X-ray interaction region using a concentric microfluidic electrokinetic sample holder (coMESH)22 operating at nominally 1 μl min −1 in each chamber; two identical, independent coMESH systems were simultaneously operated during data collection in a similar experimental set-up to that described previously21. Data for the two concurrent experiments were recorded on separate, 2.3 Megapixel Cornell-SLAC Pixel Array Detectors23 at 120 Hz operation each.…”

Section: Resultsmentioning

confidence: 99%

“…The experiments were performed using a serial operation of the serial crystallography set-up21, in which case the unused X-rays from a primary SFX experiment are refocused by Be compound refractive lenses to a second SFX experiment. Individual X-ray pulses containing ∼2 × 10 11 photons at 100% transmission with pulse duration between 35 and 40 fs were focused into a spot size of ∼3 × 3 μm in both the upstream and downstream experiments.…”

Section: Methodsmentioning

confidence: 99%

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Selenium single-wavelength anomalous diffraction de novo phasing using an X-ray-free electron laser

et al. 2016

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Structural information about biological macromolecules near the atomic scale provides important insight into the functions of these molecules. To date, X-ray crystallography has been the predominant method used for macromolecular structure determination. However, challenges exist when solving structures with X-rays, including the phase problem and radiation damage. X-ray-free electron lasers (X-ray FELs) have enabled collection of diffraction information before the onset of radiation damage, yet the majority of structures solved at X-ray FELs have been phased using external information via molecular replacement. De novo phasing at X-ray FELs has proven challenging due in part to per-pulse variations in intensity and wavelength. Here we report the solution of a selenobiotinyl-streptavidin structure using phases obtained by the anomalous diffraction of selenium measured at a single wavelength (Se-SAD) at the Linac Coherent Light Source. Our results demonstrate Se-SAD, routinely employed at synchrotrons for novel structure determination, is now possible at X-ray FELs.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Selenium single-wavelength anomalous diffraction de novo phasing using an X-ray-free electron laser

et al. 2016

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“…The 64 tiles are arranged in quadrants, positioned in such a way that the strong direct beam can pass through the hole in the center. The unused beam can then be refocused downstream, allowing for several experiments to be performed simultaneously [41]. …”

Section: Detectors For Xfelsmentioning

confidence: 99%

A Bright Future for Serial Femtosecond Crystallography with XFELs

Johansson

Stauch

Ishchenko

et al. 2017

Trends in Biochemical Sciences

131

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X-ray free electron lasers (XFELs) have potential to revolutionize macromolecular structural biology due to the unique combination of spatial coherence, extreme peak brilliance and short duration of X-ray pulses. A recently emerged serial femtosecond crystallography (SFX) approach using XFEL radiation overcomes some of the biggest hurdles of traditional crystallography related to radiation damage through the diffraction-before-destruction principle. Intense femtosecond XFEL pulses enable high-resolution room temperature structure determination of difficult to crystallize biological macromolecules, while simultaneously opening up a new era of time-resolved structural studies. Here, we review the latest developments in instrumentation, sample delivery, data analysis, crystallization methods and applications of SFX to important biological questions, and conclude with brief insights into the bright future of structural biology using XFELs.

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“…Current facilities are acutely aware of this and are doing their utmost to increase the capabilities and availability of instruments, such as offering 6-hour slots for crystal screening. The LCLS is currently constructing a beamline for in-air measurements and has also shown the viability of refocusing the spent beam in a SFX experiment (passing through the hole in the detector) to run a second SFX measurement in parallel [48]. Availability will soon increase with new facilities to be opened, such as the European XFEL, SwissFEL, and PAL in Korea, all of which have plans for crystallography stations.…”

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confidence: 99%

Serial Femtosecond Crystallography

Chapman

2015

Synchrotron Radiation News

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X-ray free-electron lasers produce brief flashes of X-rays that are of about a billion times higher peak brightness than achievable from storage ring sources. Such a tremendous jump in X-ray source capabilities, that came in 2009 when the Linac Coherent Light Source begun operations, was unprecedented in the history of X-ray science. Protein structure determination through the method of macromolecular crystallography has consistently benefited from the many increases in source performance from rotating anodes to all generations of synchrotron facilities. But when confronted with the prospects of such bright beams for structural biology, enthusiastic proposals were tempered by a trepidation of the effects of such beams on samples and challenges to record data [1]. A decade after these discussions (and others in USA) on the applications of X-ray FELs for biology, the first experiments took place at LCLS giving results that fulfilled many of the dreams of the early visionaries. In particular, the concept that diffraction representing the pristine object could be recorded before the Xray pulse completely vaporises the object was validated [2], confirming predictions [3] that established dose limits could be vastly exceeded using femtosecond-duration pulses. The first experiments illuminated a path to achieve room-temperature structures free of radiation damage, from samples too small to provide useful data at synchrotron facilities, as well as providing the means to carry out time-resolved crystallography at femtoseconds to milliseconds. In the five years since, progress has been substantial and rapid, invigorating the field of macromolecular crystallography [4,5]. This phase of development is far from over, but with both the LCLS and the Spring-8 Ångström Compact Free-electron Laser (SACLA) providing facilities for measurements, the benefits of X-ray FELs are already being translated into new biological insights.One of the main bottlenecks in protein structure determination is the need for large welldiffracting crystals of the protein of interest, which can take months or years of effort to achieve, if at all. Smaller crystals should be easier to obtain, given that showers of small crystals are often observed in crystallisation trials. For a fixed defect density, the likelihood to obtain a portion of well ordered crystals should increase as their volume decreases. The strength of a diffraction pattern of such an ideal crystal is proportional to the number of unit cells, and thus the volume, of the crystal. It is also proportional to the

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Characterization and use of the spent beam for serial operation of LCLS

Abstract: Serial operation of FEL experiments was performed using the spent beam typically discarded after passing through a hole in a detector. The beam profile was characterized and results for two simultaneous experiments are reported.

Cited by 17 publications

References 50 publications

Selenium single-wavelength anomalous diffraction de novo phasing using an X-ray-free electron laser

Selenium single-wavelength anomalous diffraction de novo phasing using an X-ray-free electron laser

A Bright Future for Serial Femtosecond Crystallography with XFELs

Serial Femtosecond Crystallography

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