1 kHz fixed-target serial crystallography using a multilayer monochromator and an integrating pixel detector

Tolstikova, A.; Levantino, Matteo; Yefanov, Oleksandr; Hennicke, V.; Fischer, P.; Meyer, Jan; Mozzanica, A.; Redford, S.; Crosas, Eva; Opara, Nadia; Barthelmeß, Miriam; Lieske, J.; Oberthüer, D.; Wator, E.; Mohácsi, István; Wulff, Michaël; Schmitt, B.; Chapman, Henry N.; Meents, A.

doi:10.1107/s205225251900914x

Cited by 36 publications

(28 citation statements)

References 46 publications

(33 reference statements)

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“…Even though our approach is far Data and refinement statistics for each deposited structural model: glucose isomerase (Glucy), lysozyme (Lzm) and thaumatin (Thau) obtained in Kapton (K) or Mylar (M) chips. from being considered as a serial crystallography methodology (Tolstikova et al, 2019), the number of crystals obtained per chip allowed us to obtain the best from the multiple datacollection merging protocols, such as those used at XFELs, while keeping the highest attainable resolution. The final statistics for the three model proteins refined with the Phenix suite (Liebschner et al, 2019) are shown in Table 2.…”

Section: Resultsmentioning

confidence: 99%

“…With the advent of ultrabright microfocus X-ray synchrotron sources (Flot et al, 2010) and femtosecond X-ray free-electron lasers (XFELs), new technologies and methodologies are in use to extract the most from these facilities and the capabilities offered to users (Spence, , 2020Tolstikova et al, 2019). One of their main advantages is that diffraction data can (again) be collected from samples at room temperature (RT), versus the standard diffraction data collection at cryogenic temperatures.…”

Section: Introductionmentioning

confidence: 99%

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Attaining atomic resolution from in situ data collection at room temperature using counter-diffusion-based low-cost microchips

Gavira

Rodríguez-Ruiz

Martínez‐Rodríguez

et al. 2020

Acta Cryst Sect D Struct Biol

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Sample handling and manipulation for cryoprotection currently remain critical factors in X-ray structural determination. While several microchips for macromolecular crystallization have been proposed during the last two decades to partially overcome crystal-manipulation issues, increased background noise originating from the scattering of chip-fabrication materials has so far limited the attainable resolution of diffraction data. Here, the conception and use of low-cost, X-ray-transparent microchips for in situ crystallization and direct data collection, and structure determination at atomic resolution close to 1.0 Å, is presented. The chips are fabricated by a combination of either OSTEMER and Kapton or OSTEMER and Mylar materials for the implementation of counter-diffusion crystallization experiments. Both materials produce a sufficiently low scattering background to permit atomic resolution diffraction data collection at room temperature and the generation of 3D structural models of the tested model proteins lysozyme, thaumatin and glucose isomerase. Although the high symmetry of the three model protein crystals produced almost complete data sets at high resolution, the potential of in-line data merging and scaling of the multiple crystals grown along the microfluidic channels is also presented and discussed.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Attaining atomic resolution from in situ data collection at room temperature using counter-diffusion-based low-cost microchips

Gavira

Rodríguez-Ruiz

Martínez‐Rodríguez

et al. 2020

Acta Cryst Sect D Struct Biol

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“…By additionally avoiding any sealing material, this results in much lower background scattering levels and makes the crystals directly accessible to ligand and substrate solutions for ligand-binding studies and time-resolved micro-diffusion experiments. The crystallization chips are furthermore compatible with the fastscanning Roadrunner goniometer, making the method ideally suited to time-resolved experiments at XFELs (Tolstikova et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies

Lieske

Cerv

Kreida

et al. 2019

Int Union Crystallogr J

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Efficient and reliable sample delivery has remained one of the bottlenecks for serial crystallography experiments. Compared with other methods, fixed-target sample delivery offers the advantage of significantly reduced sample consumption and shorter data collection times owing to higher hit rates. Here, a new method of on-chip crystallization is reported which allows the efficient and reproducible growth of large numbers of protein crystals directly on micro-patterned silicon chips for in-situ serial crystallography experiments. Crystals are grown by sitting-drop vapor diffusion and previously established crystallization conditions can be directly applied. By reducing the number of crystal-handling steps, the method is particularly well suited for sensitive crystal systems. Excessive mother liquor can be efficiently removed from the crystals by blotting, and no sealing of the fixed-target sample holders is required to prevent the crystals from dehydrating. As a consequence, `naked' crystals are obtained on the chip, resulting in very low background scattering levels and making the crystals highly accessible for external manipulation such as the application of ligand solutions. Serial diffraction experiments carried out at cryogenic temperatures at a synchrotron and at room temperature at an X-ray free-electron laser yielded high-quality X-ray structures of the human membrane protein aquaporin 2 and two new ligand-bound structures of thermolysin and the human kinase DRAK2. The results highlight the applicability of the method for future high-throughput on-chip screening of pharmaceutical compounds.

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“…In Laue crystallography, the bandwidth is large enough that many reflections sit entirely within the Ewald sphere volume in reciprocal space and are therefore fully recorded. A serial polychromatic crystallography approach has been demonstrated with "pink"-beam undulator radiation and a bandwidth of 2.5% at synchrotrons [28,29]. At XFELs, the use of increased bandwidth for serial crystallography could improve the partiality determination and postrefinement results, though structure factor amplitude noise from the spectral fluctuation intrinsic to SASE operation remains present.…”

Section: Introductionmentioning

confidence: 99%

Applications and Limits of Time-to-Energy Mapping of Protein Crystal Diffraction Using Energy-Chirped Polychromatic XFEL Pulses

et al. 2020

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A broadband energy-chirped hard X-ray pulse has been demonstrated at the SwissFEL (free electron laser) with up to 4% bandwidth. We consider the characteristic parameters for analyzing the time dependence of stationary protein diffraction with energy-chirped pulses. Depending on crystal mosaic spread, convergence, and recordable resolution, individual reflections are expected to spend at least ≈ 50 attoseconds and up to ≈ 8 femtoseconds in reflecting condition. Using parameters for a chirped XFEL pulse obtained from simulations of 4% bandwidth conditions, ray-tracing simulations have been carried out to demonstrate the temporal streaking across individual reflections and resolution ranges for protein crystal diffraction. Simulations performed at a higher chirp (10%) emphasize the importance of chirp magnitude that would allow increased observation statistics for the temporal separation of individual reflections for merging and structure determination. Finally, we consider the fundamental limitation for obtaining time-dependent observations using chirped pulse diffraction. We consider the maximum theoretical time resolution achievable to be on the order of 50–200 as from the instantaneous bandwidth of the chirped SASE pulse. We then assess the ability to propagate ultrafast optical pulses for pump-probe cross-correlation under characteristic conditions of material dispersion; in this regard, the limiting factors for time resolution scale with crystal thickness. Crystals that are below a few microns in size will be necessary for subfemtosecond time resolution.

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1 kHz fixed-target serial crystallography using a multilayer monochromator and an integrating pixel detector

Cited by 36 publications

References 46 publications

Attaining atomic resolution from in situ data collection at room temperature using counter-diffusion-based low-cost microchips

Attaining atomic resolution from in situ data collection at room temperature using counter-diffusion-based low-cost microchips

On-chip crystallization for serial crystallography experiments and on-chip ligand-binding studies

Applications and Limits of Time-to-Energy Mapping of Protein Crystal Diffraction Using Energy-Chirped Polychromatic XFEL Pulses

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