Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array

Lyubimov, A.Y.; Murray, Thomas D.; Koehl, Antoine; Araci, Ismail Emre; Uervirojnangkoorn, Monarin; Zeldin, Oliver B.; Cohen, Aina E.; Soltis, S. Michael; Baxter, E.L.; Brewster, Aaron S.; Sauter, Nicholas K.; Brunger, Axel T.; Berger, James M.

doi:10.1107/s1399004715002308

Cited by 68 publications

(73 citation statements)

References 65 publications

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“…A fixed target method is now capable of delivering crystals at the repetition rates of the existing XFEL sources [50] . Microfluidic flow cells would be useful for precious protein samples [51,52] . Sample injection with a coaxial sheath liquid can help to suppress a sample amount even in the case of continuous-flow liquid jet [34] .…”

Section: Discussionmentioning

confidence: 99%

Fluid sample injectors for x-ray free electron laser at SACLA

Tono

2017

High Pow Laser Sci Eng

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This paper provides a review on sample injectors which are provided at SPring-8 Angstrom Compact free electron LAser (SACLA) for conducting serial measurement in a 'diffract-before-destroy' scheme using an x-ray free electron laser (XFEL). Versatile experimental platforms at SACLA are able to accept various types of injectors, among which liquidjet, droplet and viscous carrier injectors are frequently utilized. These injectors produce different forms of fluid targets such as a liquid filament with a diameter in the order of micrometer, micro-droplet synchronized to XFEL pulses, and slowly flowing column of highly viscous fluid with a rate below 1 µL min −1 . Characteristics and applications of the injectors are described.

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Section: Discussionmentioning

confidence: 99%

Fluid sample injectors for x-ray free electron laser at SACLA

Tono

2017

High Pow Laser Sci Eng

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“…25,31,33,34,37,143,185,[191][192][193][194][195] The simplest mounting strategy involves the deposition of a crystal slurry between two thin silicon nitride wafers. 143 However, most approaches have utilized microfabricated mesh-type structures.…”

Section: Sample Arraysmentioning

confidence: 99%

“…Brunger et al reported a microfluidic trap array which utilized hydrodynamic forces to arrange microcrystals grown off-chip into an ordered array ( Figure 9). 33 In contrast, Fraden et al reported that kinetic optimization of droplet-microfluidics was used to ensure the growth of only a single in each droplet. 15 The droplets were then arranged into a microfluidic array for subsequent analysis (Figure 8(b)).…”

Section: Sample Arraysmentioning

confidence: 99%

“…[19][20][21][22][23][24][25][26][27][28][29][30] However, these large-scale serial methods suffer from the need to manipulate crystals and/ or from inefficient sample utilization. 3,[31][32][33][34][35][36] The material-intensive nature of these experiments extends the long-standing challenge of growing a well-diffracting crystal to reproducibly generating a large number of high quality, isomorphous crystals. These issues are then further compounded by the need to deliver those samples efficiently to the X-ray beam.…”

Section: Introductionmentioning

confidence: 99%

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Microfluidics: From crystallization to serial time-resolved crystallography

Sui

Perry

2017

Structural Dynamics

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Capturing protein structural dynamics in real-time has tremendous potential in elucidating biological functions and providing information for structure-based drug design. While time-resolved structure determination has long been considered inaccessible for a vast majority of protein targets, serial methods for crystallography have remarkable potential in facilitating such analyses. Here, we review the impact of microfluidic technologies on protein crystal growth and X-ray diffraction analysis. In particular, we focus on applications of microfluidics for use in serial crystallography experiments for the time-resolved determination of protein structural dynamics.

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“…Such platforms have been increasingly harnessed to facilitate the diffraction analysis of challenging targets for both static and dynamic structure determination. Various platforms have been developed to improve the growth and subsequent mounting of tiny and fragile crystals for X-ray diffraction analysis [24][25][26][27][28], including dense array-style devices [29][30][31][32][33][34][35][36][37][38][39][40][41], platforms for the lipidic cubic phase crystallization of membrane proteins [42,43], and thin-film sandwich devices [44]. In the meantime, the challenges of such platforms lie in the need to maintain a protected sample environment, as well as minimize the interference of device materials with the subsequent X-ray analysis.…”

Section: Introductionmentioning

confidence: 99%

A Graphene-Based Microfluidic Platform for Electrocrystallization and In Situ X-ray Diffraction

et al. 2018

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Here, we describe a novel microfluidic platform for use in electrocrystallization experiments. The device incorporates ultra-thin graphene-based films as electrodes and as X-ray transparent windows to enable in situ X-ray diffraction analysis. Furthermore, large-area graphene films serve as a gas barrier, creating a stable sample environment over time. We characterize different methods for fabricating graphene electrodes, and validate the electrical capabilities of our device through the use of methyl viologen, a redox-sensitive dye. Proof-of-concept electrocrystallization experiments using an internal electric field at constant potential were performed using hen egg-white lysozyme (HEWL) as a model system. We observed faster nucleation and crystal growth, as well as a higher signal-to-noise for diffraction data obtained from crystals prepared in the presence of an applied electric field. Although this work is focused on the electrocrystallization of proteins for structural biology, we anticipate that this technology should also find utility in a broad range of both X-ray technologies and other applications of microfluidic technology.

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Capture and X-ray diffraction studies of protein microcrystals in a microfluidic trap array

Abstract: A microfluidic platform has been developed for the capture and X-ray analysis of protein microcrystals, affording a means to improve the efficiency of XFEL and synchrotron experiments.

Cited by 68 publications

References 65 publications

Fluid sample injectors for x-ray free electron laser at SACLA

Fluid sample injectors for x-ray free electron laser at SACLA

Microfluidics: From crystallization to serial time-resolved crystallography

A Graphene-Based Microfluidic Platform for Electrocrystallization and In Situ X-ray Diffraction

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