A Double Emulsion-Based, Plastic-Glass Hybrid Microfluidic Platform for Protein Crystallization

Published on Oct 28, 2015in Micromachines2.43
· DOI :10.3390/mi6111446
Deyong Zhu2
Estimated H-index: 2
Xiaohu Zhou1
Estimated H-index: 1
Bo Zheng1
Estimated H-index: 1
This paper reports the design and construction of a plastic-glass hybrid microfluidic platform for performing protein crystallization trials in nanoliter double emulsions. The double emulsion-based protein crystallization trials were implemented with both the vapor-diffusion method and microbatch method by controlling the diffusion of water between the inner and outer phases and by eliminating water evaporation. Double emulsions, whose inner and outer environments can be easily adjusted, can provide ideal conditions to explore protein crystallization with the advantages of a convection-free environment and a homogeneous interface. The property of the water-oil interface was demonstrated to be a critical factor for nucleation, and appropriate surfactants should be chosen to prevent protein adsorption at the interface. The results from the volume effect study showed a trend of fewer crystals and longer incubation time when the protein solution volume became smaller, suggesting that the nucleation in protein crystallization process can be controlled by changing the volume of protein solutions. Finally, sparse matrix screening was achieved using the double emulsion-based microbatch method. The double emulsion-based approach for protein crystallization is a promising tool for enhancing the crystal quality by controlling the nucleation process.
  • References (66)
  • Citations (3)
Chia-Ying Huang5
Estimated H-index: 5
(Trinity College, Dublin),
Vincent Olieric14
Estimated H-index: 14
(Paul Scherrer Institute)
+ 4 AuthorsMartin Caffrey54
Estimated H-index: 54
(Trinity College, Dublin)
The lipid cubic phase (LCP) continues to grow in popularity as a medium in which to generate crystals of membrane (and soluble) proteins for high-resolution X-ray crystallographic structure determination. To date, the PDB includes 227 records attributed to the LCP or in meso method. Among the listings are some of the highest profile membrane proteins, including the β2-adrenoreceptor–Gs protein complex that figured in the award of the 2012 Nobel Prize in Chemistry to Lefkowitz and Kobilka. The mo...
Published on May 20, 2015in Journal of the American Chemical Society14.70
Dongtang Zhang5
Estimated H-index: 5
Fuxiang Wu2
Estimated H-index: 2
+ 3 AuthorsGuangsheng Guo16
Estimated H-index: 16
Ordered intermetallic nanomaterials are of considerable interest for fuel cell applications because of their unique electronic and structural properties. The synthesis of intermetallic compounds generally requires the use of high temperatures and multiple-step processes. The development of techniques for rapid phase- and size-controlled synthesis remains a formidable challenge. The intermetallic compound Pt1Bi2 is a promising candidate catalyst for direct methanol fuel cells because of its high ...
Published on May 19, 2015in Langmuir3.68
Jana Bahtz2
Estimated H-index: 2
Deniz Z. Gunes10
Estimated H-index: 10
+ 5 AuthorsErich J. Windhab31
Estimated H-index: 31
This contribution reports on the mass transport kinetics of osmotically imbalanced water-in-oil-in-water (W1/O/W2) emulsions. Although frequently studied, the control of mass transport in W1/O/W2 emulsions is still challenging. We describe a microfluidics-based method to systematically investigate the impact of various parameters, such as osmotic pressure gradient, oil phase viscosity, and temperature, on the mass transport. Combined with optical microscopy analyses, we are able to identify and ...
Published on May 1, 2015in Scientific Reports4.01
Ying Zhu14
Estimated H-index: 14
Li-Na Zhu2
Estimated H-index: 2
+ 3 AuthorsQun Fang26
Estimated H-index: 26
Large-scale screening of hundreds or even thousands of crystallization conditions while with low sample consumption is in urgent need, in current structural biology research. Here we describe a fully-automated droplet robot for nanoliter-scale crystallization screening that combines the advantages of both automated robotics technique for protein crystallization screening and the droplet-based microfluidic technique. A semi-contact dispensing method was developed to achieve flexible, programmable...
Published on Mar 1, 2015in Plant Cell and Environment5.62
Zhengjiang Qian2
Estimated H-index: 2
(CAS: Chinese Academy of Sciences),
Juan‐Juan Song2
Estimated H-index: 2
(CAS: Chinese Academy of Sciences)
+ 1 AuthorsQing Ye15
Estimated H-index: 15
(CAS: Chinese Academy of Sciences)
It has long been recognized that inhibition of plant water transport by either osmotic stress or salinity is mediated by aquaporins (AQPs), but the function and regulation of AQPs are highly variable among distinct isoforms and across different species. In this study, cucumber seedlings were subjected to PEG or NaCl stress for duration of 2h or 24h. The 2h treatment with PEG or NaCl had non-significant effect on the expression of plasma membrane AQP (CsPIPs) in roots, indicating the decrease in ...
Published on Sep 3, 2014in Crystal Growth & Design4.15
Sathish V. Akella4
Estimated H-index: 4
Aaron Mowitz1
Estimated H-index: 1
+ 1 AuthorsSeth Fraden36
Estimated H-index: 36
We measured the nucleation rates of lysozyme protein crystals using microfluidically produced emulsion drops containing supersaturated protein solution. The technique involves quenching several thousand independent nanoliter drops by rapidly lowering the temperature and then counting the number of drops that have not nucleated as a function of time at constant temperature. We fit the number distribution to a theoretical model developed by Pound and La Mer (J. Am. Chem. Soc. 1952, 74, 2323–2332) ...
Published on Sep 1, 2014in IUCrJ4.76
Michael Heymann14
Estimated H-index: 14
(Brandeis University),
Achini Opthalage1
Estimated H-index: 1
(Brandeis University)
+ 4 AuthorsSeth Fraden36
Estimated H-index: 36
(Brandeis University)
An emulsion-based serial crystallographic technology has been developed, in which nanolitre-sized droplets of protein solution are encapsulated in oil and stabilized by surfactant. Once the first crystal in a drop is nucleated, the small volume generates a negative feedback mechanism that lowers the supersaturation. This mechanism is exploited to produce one crystal per drop. Diffraction data are measured, one crystal at a time, from a series of room-temperature crystals stored on an X-ray semi-...
Published on Jul 9, 2014in Nano Letters12.28
Lei Zhang18
Estimated H-index: 18
Yi Wang3
Estimated H-index: 3
+ 1 AuthorsYounan Xia174
Estimated H-index: 174
Droplet reactors have received considerable attention in recent years as an alternative route to the synthesis and potentially high-volume production of colloidal metal nanocrystals. Interfacial adsorption will immediately become an important issue to address when one seeks to translate a nanocrystal synthesis from batch reactors to droplet reactors due to the involvement of higher surface-to-volume ratios for the droplets and the fact that nanocrystals tend to be concentrated at the water–oil i...
Published on Mar 28, 2014in Science41.04
Werner Kühlbrandt61
Estimated H-index: 61
(MPG: Max Planck Society)
Precise knowledge of the structure of macromolecules in the cell is essential for understanding how they function. Structures of large macromolecules can now be obtained at near-atomic resolution by averaging thousands of electron microscope images recorded before radiation damage accumulates. This is what Amunts et al. have done in their research article on page 1485 of this issue ( 1 ), reporting the structure of the large subunit of the mitochondrial ribosome at 3.2 A resolution by electron c...
Published on Mar 1, 2014in IUCrJ4.76
Cornelius Gati17
Estimated H-index: 17
G. Bourenkov13
Estimated H-index: 13
(EMBL-EBI: European Bioinformatics Institute)
+ 11 AuthorsMichael Duszenko32
Estimated H-index: 32
(University of Tübingen)
Crystal structure determinations of biological macromolecules are limited by the availability of sufficiently sized crystals and by the fact that crystal quality deteriorates during data collection owing to radiation damage. Exploiting a micrometre-sized X-ray beam, high-precision diffractometry and shutterless data acquisition with a pixel-array detector, a strategy for collecting data from many micrometre-sized crystals presented to an X-ray beam in a vitrified suspension is demonstrated. By c...
Cited By3
Published on Jun 1, 2017in Journal of Drug Delivery Science and Technology2.61
Zexia Luo1
Estimated H-index: 1
(USTC: University of Science and Technology of China),
Gang Zhao12
Estimated H-index: 12
(USTC: University of Science and Technology of China)
+ 2 AuthorsZhiquan Shu1
Estimated H-index: 1
(WSU: Washington State University)
Abstract Microfluidics is an emerging technology for synthesis of drug-loaded micro- and nano capsules for targeted delivery and controlled release, which has extensive applications in tumor therapy and pharmacological study in healthcare. A number of polydimethylsiloxane (PDMS) microfluidic devices have been developed and widely used, while PDMS based microstructure device does not support long-term stability and repeated use, moreover it requires special facilities for fabrication and plasma b...
Published on Feb 10, 2017in Micromachines2.43
Chang Nong Lim1
Estimated H-index: 1
(University of Nottingham Malaysia Campus),
Kai Seng Koh4
Estimated H-index: 4
+ 3 AuthorsYuying Yan27
Estimated H-index: 27
(UNNC: The University of Nottingham Ningbo China)
This paper demonstrates a low cost fabrication approach for microscale droplet fission and encapsulation. Using a modified xurography method, rapid yet reliable microfluidic devices with flexible designs (single layer and double layer) are developed to enable spatial control of droplet manipulation. In this paper, two different designs are demonstrated, i.e., droplet fission (single layer) and droplet encapsulation (double layer). In addition, the current fabrication approach reduces the overall...
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