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Mixing in microscale flows, where turbulence is inherently difficult to generate, has been a challenging issue owing to its laminar flow characteristics. Either the diffusion-based mixing process, or the convective mixing based on the cross-stream secondary flow, has been exploited as a passive mixing scheme that does not require any external force field. However, these techniques suffer from insufficient mixing or complicated channel design step. In this study, we propose an efficient mixing scheme by combining inertio-elastic flow instability in a viscoelastic dilute polymer solution and a modified serpentine channel, termed a gear-shape channel, which has side wells along the serpentine channel. We achieved highly efficient mixing in the gear-shaped channel for a significantly wider range of flow rates than in a conventional serpentine channel. Further, we applied our novel mixing scheme to the continuous synthesis of silica nanoparticles, which demonstrated the synthesis of nanoparticles with more uniform size distribution and regular shape, than those in a Newtonian fluid. In addition, the adsorption of inorganic materials on the channel walls was significantly suppressed by the flow instability of the viscoelastic dilute polymer solution in the gear-shaped channel.

Although cellular secretion is important in industrial biotechnology, its assessment is difficult due to the lack of efficient analytical methods. This study describes a synthetic cellular communication-based microfluidic platform for screening strains with the improved secretion of 3-hydroxypropionic acid (3-HP), an industry-relevant platform chemical. 3-HP-secreting cells were compartmentalized in droplets, with receiving cells equipped with a genetic circuit that converts the 3-HP secretion level into an easily detectable signal. This platform was applied to identify Escherichia coli genes that enhance the secretion of 3-HP. As a result, two genes (setA, encoding a sugar exporter, and yjcO, encoding a Sel1 repeat-containing protein) found by this platform enhance the secretion of 3-HP and its production. Given the increasing design capability for chemical-detecting cells, this platform has considerable potential in identifying efflux pumps for not only 3-HP but also many important chemicals.

We present a programmable microfluidic static droplet array (SDA) device that can perform user-defined multistep combinatorial protocols. It combines the passive storage of aqueous droplets without any external control with integrated microvalves for discrete sample dispensing and dispersion-free unit operation. The addressable picoliter-volume reaction is systematically achieved by consecutively merging programmable sequences of reagent droplets. The SDA device is remarkably reusable and able to perform identical enzyme kinetic experiments at least 30 times via automated cross-contamination-free removal of droplets from individual hydrodynamic traps. Taking all these features together, this programmable and reusable universal SDA device will be a general microfluidic platform that can be reprogrammed for multiple applications.

Economic production of chemicals from microbes necessitates development of high-producing strains and an efficient screening technology is crucial to maximize the effect of the most popular strain improvement method, the combinatorial approach. However, high-throughput screening has been limited for assessment of diverse intracellular metabolites at the single-cell level. Herein, we established a screening platform that couples a microfluidic static droplet array (SDA) and an artificial riboswitch to analyse intracellular metabolite concentration from single microbial cells. Using this system, we entrapped single Escherichia coli cells in SDA to measure intracellular l-tryptophan concentrations. It was validated that intracellular l-tryptophan concentration can be evaluated by the fluorescence from the riboswitch. Moreover, high-producing strains were successfully screened from a mutagenized library, exhibiting up to 145% productivity compared to its parental strain. This platform will be widely applicable to strain improvement for diverse metabolites by developing new artificial riboswitches.

Droplet-based microfluidics enabling exquisite liquid-handling has been developed for diagnosis, drug discovery and quantitative biology. Compartmentalization of samples into a large number of tiny droplets is a great approach to perform multiplex assays and to improve reliability and accuracy using a limited volume of samples. Despite significant advances in microfluidic technology, individual droplet handling in pico-volume resolution is still a challenge in obtaining more efficient and varying multiplex assays. We present a highly addressable static droplet array (SDA) enabling individual digital manipulation of a single droplet using a microvalve system. In a conventional single-layer microvalve system, the number of microvalves required is dictated by the number of operation objects; thus, individual trap-and-release on a large-scale 2D array format is highly challenging. By integrating double-layer microvalves, we achieve a “balloon” valve that preserves the pressure-on state under released pressure; this valve can allow the selective releasing and trapping of 7200 multiplexed pico-droplets using only 1 μL of sample without volume loss. This selectivity and addressability completely arranged only single-cell encapsulated droplets from a mixture of droplet compositions via repetitive selective trapping and releasing. Thus, it will be useful for efficient handling of miniscule volumes of rare or clinical samples in multiplex or combinatory assays, and the selective collection of samples.

This study demonstrates a simple approach for fabricating a 3D-μPAD from a single sheet of paper by double-sided printing and lamination. First, a wax printer prints vertically symmetrical and asymmetrical wax patterns onto a double-sided paper surface. Then, a laminator melts the printed wax patterns to form microfluidic channels in the paper sheet. The vertically symmetrical wax patterns form vertical channels when the melted wax patterns make contact with each other. The asymmetrical wax patterns form lateral and vertical channels at the cross section of the paper when the printed wax patterns are melted to a lower height than the thickness of the single sheet of paper. Finally, the two types of wax patterns form a 3D microfluidic network to move fluid laterally and vertically in the single sheet of paper. This method eliminates major technical hurdles related to the complicated and tedious alignment, assembly, bonding, and punching process. This 3D-μPAD can be used in a multiplex digital assay to measure the concentration of a target analyte in a sample solution simply by counting the number of colored bars at a fixed time. It does not require any external instruments to perform digital measurements. Therefore, we expect that this approach could be an instrument-free assay format for use in developing countries.

We demonstrate a practical design and integration of multidirectional tilted UV lithography and microinjection molding for microstructure-based microfluidic devices. The previously reported undercut (or T-profile) problem of photoresist causes the imperfect mirror image duplication of the microstructures to the Ni mold. This issue inevitably leads to the unstable molds in the production of microstructure-based microfluidic devices. This study presents a simple route for the successful fabrication of microstructure-based microfluidic devices by multidirectional tilted UV lithography. By changing slope angles of microstructures through tilting the chuck during the UV exposure, the slope angles (up to 30°) around the microstructures allow the effective prevention of the undercut problems of photoresist and facilitate easy releasing of the device with high durability. This technique can be easily used for the production of the microstructure-based microfluidic devices in microanalysis and lab-on-chip applications.

This study presents a simple microfluidic approach to the rapid fabrication of complex-shaped microfibers (e.g., single hollow, double hollow, and microbelt), with highly uniform structures, based on a combination of the spontaneous formation of polymeric jet streams and in situ photopolymerization. Two laminar flows of a photocurable fluid and a liquid template (nonpolymerizing fluid) spontaneously form jet streams in equilibrium states in microfluidic channels because of the minimization of the interfacial energy between the two fluids. The formation of the jet streams strongly depends on the spreading coefficients and the evolution time along the downstream of the microfluidic system. Thus, the simple control of the spreading coefficients can guide microfibers into various shapes. The sizes of the core and shell of the hollow fibers can also be readily manipulated by the flow rates of the polymerizing fluid and the liquid template phase. Asymmetric hollow fibers can also be produced in different evolutionary states in the microfluidic system. The microfluidic approach shown here represents a significant step toward the easy fabrication of microfibers with readily controllable structures and geometries. We anticipate that this novel fabrication approach and the prediction method based on spreading coefficients presented in this work can be applied to produce a wide variety of functional microfibrous materials.

Antibiotic resistance of biofilms is a growing public health concern due to overuse and improper use of antibiotics. Thus, determining an effective minimal concentration of antibiotics to eradicate bacterial biofilms is crucial. Here we present a simple, novel one-pot assay for the analysis of antibiotic susceptibility of bacterial biofilms using a microfluidics system where continuous concentration gradients of antibiotics are generated. The results of minimal biofilm eradication concentration (MBEC) clearly confirm that the concentration required to eradicate biofilm-grown Pseudomonas aeruginosa is higher than the minimal inhibitory concentration (MIC) that has been widely used to determine the lowest concentration of antibiotics against planktonically grown bacteria.

For cell-based biosensor applications, dissociated neurons have been cultured on planar microelectrode arrays (MEAs) to measure the network activity with substrate-embedded microelectrodes. There has been a need for a multi-well type platform to reduce the data collection time and increase the statistical power for data analysis. This study presents a novel method to convert a conventional MEA into a multi-well MEA with an array of micrometre-sized neuronal culture (‘neuronal micro-circuit array’). An MEA was coated first with cell-adhesive layer (poly-D-lysine) which was subsequently patterned with a cell-repulsive layer (agarose hydrogel) to both pattern the cell adhesive region and isolate neuronal micro-circuits from each other. For a few weeks, primary hippocampal neurons were cultured on the agarose microwell MEA and the development of spontaneous electrical activities were characterized with extracellular action potentials. Using neurotransmission modulators, the simultaneous monitoring of drug responses from neuronal micro-circuit arrays was also demonstrated. The proposed approach will be powerful for neurobiological functional assay studies or neuron-based biosensor fields which require repeated trials to obtain a single data point due to biological variations.

We present a simple synthetic approach for the preparation of cell attachable Janus polyurethane (PU) microfibers in a microfluidic system. The synthesis was performed by using laminar flows of multiple streams with spontaneous formation of carbon dioxide bubbles resulting in an asymmetrically porous PU microfiber. The fabricated asymmetric microfiber (Janus microfiber) provides two distinctive properties: one is a porous region to promote the cellular adhesion and the other is a nonporous region rendering the mechanical strength of the scaffold. The Janus microfibers show dramatic improvement of cell adhesion, proliferation, and viability over a culture period. Cells cultured on the fibers easily bridged gaps between microfibers by joining together to form a cell sheet. The maximum distance between fibers that fibroblasts bridged is approximately 200 microm over 15 days. The Janus microfiber can be used for not only an alternative 2D cell culture plate but also as a novel 3D scaffold for tissue engineering without any need for elegant surface modification for enhancing cell adhesions.

The surface of mammalian cells is densely coated with complex glycans, which are directly involved in cell-cell or cell-protein interactions that trigger various biological responses. Here, we present a novel glycomics approach that uses quantum dot (Qdot)-lectin nanoconjugates to interrogate the surface glycans of tissues and patterned cells. Our approach allows highly sensitive in situ monitoring of specific lectin-glycan interactions and quantitative information on surface glycans for each examined cell line and tissue. The results clearly show significant changes in glycosylation for each cell line and tissue sample. We expect that these results will be applicable in cancer diagnostics and promote the development of new analytical tools for glycomics.