Microfluidic chips for the crystallization of biomacromolecules by counter-diffusion and on-chip crystal X-ray analysis

Dhouib, Kaouthar; Malek, Chantal Khan; Pfleging, Wilhelm; Gauthier‐Manuel, Bernard; Duffait, Roland; Thuillier, G.; Ferrigno, Rosaria; Jacquamet, Lilian; Ohana, Jérémy; Ferrer, Jean‐Luc; Théobald-Dietrich, Anne; Giegé, Richard; Lorber, Bernard; Sauter, Claude

doi:10.1039/b819362b

Cited by 108 publications

(124 citation statements)

References 40 publications

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“…Key to this approach is the coupling of highly automated instrumentation with specialized sample containers and customized software for efficient data collection with minimal sample consumption. High-density sample containers, such as microfluidic chips or microcrystal traps (25) for room temperature studies or grids for experiments at cryogenic temperatures, hold samples in known locations. These sample holders enable very rapid and precise positioning of crystals into the X-ray interaction region for consistent production of diffraction patterns.…”

Section: Significancementioning

confidence: 99%

Goniometer-based femtosecond crystallography with X-ray free electron lasers

Cohen

Soltis

González

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

127

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The emerging method of femtosecond crystallography (FX) may extend the diffraction resolution accessible from small radiationsensitive crystals and provides a means to determine catalytically accurate structures of acutely radiation-sensitive metalloenzymes. Automated goniometer-based instrumentation developed for use at the Linac Coherent Light Source enabled efficient and flexible FX experiments to be performed on a variety of sample types. In the case of rod-shaped Cpl hydrogenase crystals, only five crystals and about 30 min of beam time were used to obtain the 125 still diffraction patterns used to produce a 1.6-Å resolution electron density map. For smaller crystals, high-density grids were used to increase sample throughput; 930 myoglobin crystals mounted at random orientation inside 32 grids were exposed, demonstrating the utility of this approach. Screening results from cryocooled crystals of β 2 -adrenoreceptor and an RNA polymerase II complex indicate the potential to extend the diffraction resolution obtainable from very radiation-sensitive samples beyond that possible with undulator-based synchrotron sources.femtosecond diffraction | crystallography | XFEL | structural biology U sing extremely bright, short-timescale X-ray pulses produced by X-ray free-electron lasers (XFELs), femtosecond crystallography (FX) is an emerging method that expands the structural information accessible from very small or very radiation-sensitive macromolecular crystals. Central to this method is the "diffraction before destruction" (1) process in which a still diffraction image is produced by a single X-ray pulse before significant radiation-induced electronic and atomic rearrangements occur within the crystal (1-3). At the Linac Coherent Light Source (LCLS) at SLAC, a single ∼50-fs-long X-ray pulse can expose a crystal to as many X-ray photons as a typical synchrotron beam line produces in about a second. Exposing small crystals to these intense ultrashort pulses circumvents the dose limitations of conventional X-ray diffraction experiments (4) and may produce useful data to resolutions beyond what is achievable at synchrotrons (5). This innovation provides a pathway to obtain atomic information from proteins that only form micrometer-to nanometer-sized crystals, such as many membrane proteins and large multiprotein complexes. Moreover, XFELs enable "diffraction before reduction" data collection to address another major challenge in structural enzymology by providing a means to determine catalytically accurate structures of acutely radiation-sensitive metalloenzyme active sites (6), such as high-valency reaction intermediates that may be significantly photoreduced during a single X-ray exposure at a synchrotron, even at very small doses (7-11). Furthermore, the use of short (tens of femtoseconds) X-ray pulses further complements the structural characterization of biochemical reaction processes by providing access to a time domain two to three orders of magnitude faster (12, 13) than currently accessible using synchrotro...

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Section: Significancementioning

confidence: 99%

Goniometer-based femtosecond crystallography with X-ray free electron lasers

Cohen

Soltis

González

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

127

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“…To minimize manual handling, an alternative is in situ X-ray data collection. One method involves using X-ray-transparent microfluidic devices (Hansen et al, 2006, Dhouib et al, 2009, Stojanoff et al, 2011, Guha et al, 2012, Pinker et al, 2013, Khvostichenko et al, 2014, Horstman et al, 2015, Heymann et al, 2015, Maeki et al, 2015. Another solution following Yadav's pioneering work (Yadav et al, 2005), is to collect X-ray data directly in micro capillary (Li et al, 2006, Maeki et al, 2012.…”

Section: Introductionmentioning

confidence: 99%

Crystallization via tubing microfluidics permits both in situ and ex situ X-ray diffraction

Gerard¹,

Ferry²,

Vuillard³

et al. 2017

Acta Cryst Sect F

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“…Micro injection moulding (µ-IM) processes are highly suited for high volume manufacture of these devices which are inexpensive and light weight, and can be considered as disposable alternatives to ceramic platforms. Cyclic olefin copolymer (COC) is one of the polymers that are suitable for producing microfluidics parts because of its high glass transition temperatures (T g ), low moisture uptake, high chemical resistance, excellent optical properties, bio-compatibility, sterilization possibility and thus suitability for medical approval [4,5,6].…”

Section: Introductionmentioning

confidence: 99%

Characterisation of demoulding parameters in micro-injection moulding

et al. 2014

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Condition monitoring of micro injection moulding is an effective way of understanding the processing effects of variable parameter settings. This paper reports an experimental study that investigates the characteristics of the demoulding behaviour in micro injection moulding (µ-IM) with a focus on the process factors that affect parts' quality. Using a Cyclic Olefin Copolyme (COC) microfluidics demonstrator, the demoulding performance was studied as a function of four process parameters (melt temperature, mould temperature, holding pressure and injection speed), employing the design of experiment approach. The results provide empirical evidences on the effect that processing parameters have on demoulding conditions in µ-IM, and identifies combinations of parameters that can be used to achieve the optimal processing conditions in regards to demoulding behaviour of micro parts. It was concluded that there was a direct correlation between the applied pressure during part filling, holding phases and the demoulding characteristic factors of the µ-IM cycle such as ejection force, integral and time.2

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Microfluidic chips for the crystallization of biomacromolecules by counter-diffusion and on-chip crystal X-ray analysis

Cited by 108 publications

References 40 publications

Goniometer-based femtosecond crystallography with X-ray free electron lasers

Goniometer-based femtosecond crystallography with X-ray free electron lasers

Crystallization via tubing microfluidics permits both in situ and ex situ X-ray diffraction

Characterisation of demoulding parameters in micro-injection moulding

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