An attempt was made to arrange polystyrene latex particles (2, 5, and 10 microm in diameter) dispersed in aqueous media making use of their dielectrophoresis and electrophoresis with a hyperbolic quadruple electrode system. Application of a high-frequency ac field enabled the particles to arrange themselves between the electrodes forming a particle monolayer due to the negative dielectrophoretic force. Simultaneous application of high-frequency ac and dc fields caused the particles to gather in the region surrounded by the electrodes to form particle multilayers. Appropriate choice of the way of applying an electric field thus allowed the reversible control of particle arrangements (monolayer, multilayer, dispersion). Reapplication of an ac field to the particle layers produced highly dense particle multilayers.

Herein, we demonstrate the potential of droplet-based microfluidics for controlling protein crystallization and generating single-protein crystals. We estimated the critical droplet size for obtaining a single crystal within a microdroplet and investigated the crystallization of four model proteins to confirm the effect of protein molecular diffusion on crystallization. A single crystal was obtained in microdroplets smaller than the critical size by using droplet-based microfluidics. In the case of thaumatin crystallization, a single thaumatin crystal was obtained in a 200 μm droplet even with high supersaturation. In the case of ferritin crystallization, the nucleation profile of ferritin crystals had a wider distribution than the nucleation profiles of lysozyme, thaumatin, and glucose isomerase crystallization. We found that the droplet-based microfluidic approach was able to control the nucleation of a protein by providing control over the crystallization conditions and the droplet size, and that the diffusion of protein molecules is a significant factor in controlling the nucleation of protein crystals in droplet-based microfluidics.

We fabricated a simple microfluidic device for separation of bovine oocytes based on the oocyte quality to improve the conception rate of in vitro fertilization (IVF) by using good quality oocytes. The microfluidic device separates oocytes based on sedimentation rate differences in a sucrose buffer, which is dependent on oocyte quality. The microfluidic device has a 700 µm width, 1 mm height, and 10 mm long separation channel. Oocytes were injected from the upper half of the separation channel, and they flowed while sinking. The outlets of the separation channel were divided into upper and lower chambers. Good quality oocytes settled faster than poor quality oocytes in sucrose buffer; therefore, good quality oocytes were collected from the lower outlet. We performed IVF after the microfluidic separation of oocytes. The developmental rate to blastocysts of oocytes collected from the lower outlet was significantly higher than those collected from the upper outlet (36.0% vs. 14.1%). This result was comparable to that in the BCB staining method performed as a comparison method (BCB+ : 35.7%, BCB−: 15.4%). These findings indicate that our microfluidic device could be applied to oocyte separation and contribute to improvement of in vitro embryo production system.

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We investigate the behaviour of a simple microfluidic device designed to separate particles based on density. The device consists of a separation-channel with three inlet and two outlet channels. The particle samples were loaded in the middle of the main channel. The results of separation experiments using model particles provide clear evidence that this approach can be used to achieve good separation of particles of close density populations. Particle separation was found more favourable with low flow rates. Forces exerted on particles were modelled by Stokes' law and found to be consistent with experimental results. One advantage of this microfluidic system is low exposure of the sample to hydrodynamic shear stresses compared with conventional methods. This device may be adopted to precisely handle single cells and easily interface with other tools and separation techniques.

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We developed four microsatellite DNA loci to test for multiple paternity of black rockfish, Sebastes inermis, from the Seto Inland Sea of Japan. All loci showed a high degree of polymorphism (number of alleles per locus = 10-14, expected heterozygosity = 0.80) and discriminating power (probability of identity index = 3.71 9 10 -6 , exclusion probability = 0.999) in unrelated wild specimens (n = 32). Genotypic assignment of five dams (109-220 mm in total length) and 50 embryos from each dam (n = 50) indicated that four dams were mated with a single sire. Only for one dam and three of her embryos we could not exclude multiple paternity.

… could be formed in a controlled manner by using droplet-based microfluidics. In the Full Paper on page 1049 ff., M. Miyazaki and co-workers show that this method was able to direct the nucleation profile of a protein by providing control over the crystallization conditions and the droplet size. The diffusion of protein molecules played an important role in the nucleation of protein crystals.