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We developed a method for passively controlling microdroplet rotation, including interior rotation, using a parallel flow comprising silicone and sesame oils. This device has a simple 2D structure with a straight channel and T-junctions fabricated from polydimethylsiloxane. A microdroplet that forms upstream moves into the sesame oil. Then, the largest flow velocity at the interface of the two oil layers applies a rotational force to the microdroplet. A microdroplet in the lower oil rotates clockwise while that in the upper oil rotates anti-clockwise. The rotational direction was controlled by a simple combination of sesame and silicone oils. Droplet interior flow was visualized by tracking microbeads inside the microdroplets. This study will contribute to the efficient creation of chiral molecules for pharmaceutical and materials development by controlling rotational direction and speed.

For high throughput size controllable micro droplet generation, a three step half and half size/volume division of source droplet device using three stage cascade channel is introduced. Passive and active size/volume ratio controllable source droplet division devises are reported next. Finally, two types of microdroplet (daughter) generation devices using hydrodynamic droplet breakup of source droplet (mother) are described.

In recent years, research on the application of microdroplets in the fields of biotechnology and chemistry has made remarkable progress, but the technology for the stable generation of single-micrometer-scale microdroplets has not yet been established. In this paper, we developed an efficient and stable single-micrometer-scale droplet generation device based on the fragmentation of droplet tails, called “tail thread mode”, that appears under moderate flow conditions. This method can efficiently encapsulate microbeads that mimic cells and chemical products in passively generated single-micrometer-scale microdroplets. The device has a simple 2D structure, a T-junction is used for droplet generation, and in the downstream, multi-branch channels are designed for droplet deformation into the tail. Several 1–2 µm droplets were successfully produced by the tail’s fragmentation, this continuous splitting was induced by the branch channels. We examined a wide range of experimental conditions and found the optimal flow rate condition can be reduced to one-tenth compared to the conventional tip-streaming method. A mold was fabricated by simple soft lithography, and a polydimethylsiloxane (PDMS) device was fabricated using the mold. Based on the 15 patterns of experimental conditions and the results, the key factors for the generation of microdroplets in this device were examined. In the most efficient condition, 61.1% of the total droplets generated were smaller than 2 μm.

Bacterial persisters are phenotypic variants that survive the treatment of lethal doses of growth-targeting antibiotics without mutations. Although the mechanism underlying persister formation has been studied for decades, how the persister phenotype is switched on and protects itself from antibiotics has been elusive. In this study, we focused on the lactate dehydrogenase gene (ldhA) that was upregulated in an Escherichia coli persister-enriched population. A survival rate assay using an ldhA-overexpressing strain showed that ldhA expression induced persister formation. To identify ldhA-mediated persister formation at the single-cell level, time-lapse microscopy with a microfluidic device was used. Stochastic ldhA expression was found to induce dormancy and tolerance against high-dose ampicillin treatment (500 μg/ml). To better understand the underlying mechanism, we investigated the relationship between ldhA-mediated persister formation and previously reported persister formation through aerobic metabolism repression. As a result, ldhA expression enhanced the proton motive force (PMF) and ATP synthesis. These findings suggest that ldhA-mediated persister formation pathway is different from previously reported persister formation via repression of aerobic metabolism.

We fabricated a microfluidic device with a three-dimensional (3D) structure and verified its droplet-generation performance for the stable production of droplets of around 10 μm in size. We compared the performance of the 3D device with that of conventional simple T-junction and cross-junction structures. The continuous phase sheared the dispersed phase into droplets from eight directions in the 3D device, compared with only one direction in the T-junction device and two in the cross-junction device. Droplets were produced efficiently over a wide range of fluid properties and flow conditions with the 3D device, unlike with the two conventional planar devices. Fluidic experiments were conducted using mineral oil with a surfactant as the continuous phase, deionized (DI) water as the dispersed phase, and DI water with glycerin to change the viscosity of the dispersed phase. The minimum droplet length was 47.2 μm in the T-junction device, 39.0 μm in the cross-junction device, and 22.4 μm in the 3D device when using a water and glycerin mixture with a viscosity of 9.0 mPa·s. Compared with the conventional devices, smaller droplets were produced using our 3D device, indicating that it has excellent droplet-generation performance.

This study describes the synthesis of an azo-Mn(II) complex requiring an accurate pH control. The reaction conditions for the delicate azo-Mn(II) complex could be precisely controlled using a microfluidic device. The microfluidic method showed the following advantages over the conventional method: the concentration of the pH-control reagent was reduced (1/15), the reaction time was remarkably decreased from 4 h to less than 1 s, and the reaction temperature was lowered from 40 to 23 °C. Moreover, the microfluidic device suppressed the oxidation of the compound and did not require cooling to remove the heat of reaction. In addition, the conventional method uses harsh pH control, whereas the microfluidic device permits mild pH control.

This paper presents the formation of complex cross-sectional microfibers using three-dimensional microfluidic devices. The compartments and shapes of core and shell layers in the microfibers were independently controlled via three-dimensional fluidic channels fabricated by the combination of sheath units. The number of layers is easily expanded by the stacking of these units. Therefore, the highly heterogeneous microfibers of alginate hydrogel are obtained in polydimethylsiloxane structures. This widely expandable method has great potential for the development of functional and complex fiber-shaped materials.

A fully passive volume-dependent droplet sorter is presented in this research. Repeated and multiple on-rail transfer of microdroplets in a cascade channel resulted in high resolution sorting of droplet-phase samples. Microdroplets with volumes of 0.89–3.26 nl were successfully sorted in eight steps, and a maximum sorting resolution of about 0.21 nl was obtained over three transfer stages. Line rails and dot rails employed in the cascading channels allowed volume-dependent droplet transfer, and shape recovery of the transferred droplet for the repeatable transfer, respectively. This simple device and method can be applied to droplet-based chemical or biological research.

This paper presents a method for utilizing three-dimensional microfluidic channels fully to realize multiple functions in a single device. The final device structure was achieved by combining three independent modules that consisted of horizontal and vertical channels. The device allowed for the one-step generation of water-in-oil-in-water droplets without the need for partial treatment of the polydimethylsiloxane channel surface using separate modules for generating water-in-oil droplets on the horizontal plane and oil-in-water droplets on the vertical plane. The second vertically structured module provided an efficient flow for the generation of highly wettable liquid droplets, and tuning of the first horizontally structured module enabled different modes of inner-core encapsulation within the oil shell. The successful integration of the vertical and horizontal channels for core-shell droplet generation and the chemical synthesis of a metal complex within the droplets were evaluated. The proposed approach of integrating independent modules will expand and enhance the functions of microfluidic platforms.

In a microfluidic device developed using a Si micro-pillar array directly coated with an iron catalyst, 4,8-dibromo-1-hydroxy-2-naphthaldehyde was catalytically synthesized with high efficiency. This represents the first dibromination of the organic compound in a microfluidic device. Typical beaker synthesis of the compound takes one day at 50 °C, but the method proposed here was performed at uncontrolled room temperature (23 °C), while reducing the reaction time to about 1 sec.

A simple printing process to design a novel piezoelectric energy-harvesting device made from vinylidene fluoride/trifluoroethylene copolymers is presented. Fabrication using a metal nanoink and a household printer dramatically reduced the fabrication complexity. In addition, this process employed low-temperature steps, and thus the structural damage to the polymers resulting from high-temperature annealing was avoided. By stacking the devices and connecting them in parallel, the generated energy was increased and electric power of ∼1.12 μJ was obtained.

This study reports a fast and quantitative determination method for phenylalanine (Phe) and tyrosine (Tyr) in human plasma using on-chip pressure-driven liquid chromatography. A pillar array column with low-dispersion turns and a gradient elution system was used. The separation of fluorescent derivatives of Phe, Tyr, and other hydrophobic amino acids was successfully performed within 140 s. Under the optimized conditions, Phe and Tyr in human plasma were quantified. The developed method is promising for rapid diagnosis in the clinical field.

Efficient microfluidic synthesis of chiral salen Mn(II) and Co(II) complexes containing lysozyme was achieved. The reaction conditions for the delicate protein were controlled precisely with a separation barrier in the microfluidic device. The microfluidic method has the following advantages over the conventional method: a remarkable reduction in the reaction time from 4.5 h to less than 1 s for the synthesis of Mn(II) and Co(II) complexes, a reduction in the reaction temperature from 40 to 23 °C, and no need for an N2 atmosphere because all the reagents are isolated from the air. A three-fold yield improvement was achieved for Mn(II) and Co(II) complexes containing lysozyme compared with conventional synthesis.

Recently, chemical operations with microfluidic devices, especially droplet-based operations, have attracted considerable attention because they can provide an isolated small-volume reaction field. However, analysis of these operations has been limited mostly to aqueous-phase reactions in water droplets due to device material restrictions. In this study, we have successfully demonstrated droplet formation of five common organic solvents frequently used in chemical synthesis by using a simple silicon/glass-based microfluidic device. When an immiscible liquid with surfactant was used as the continuous phase, the organic solvent formed droplets similar to water-in-oil droplets in the device. In contrast to conventional microfluidic devices composed of resins, which are susceptible to swelling in organic solvents, the developed microfluidic device did not undergo swelling owing to the high chemical resistance of the constituent materials. Therefore, the device has potential applications for various chemical reactions involving organic solvents. Furthermore, this droplet generation device enabled control of droplet size by adjusting the liquid flow rate. The droplet generation method proposed in this work will contribute to the study of organic reactions in microdroplets and will be useful for evaluating scaling effects in various chemical reactions.

In this work, an axial-type vibration energy harvester with an impact-sliding structure that uses a magnetostrictive Permendur (Fe-Co-V) rod was produced and its power generation properties were measured. It was found that all the peak frequencies analyzed using the FFT method from the experimentally obtained voltage waveforms were consistent with the calculated inherent frequencies of the axial vibration of the rod. Therefore, the power generation by the proposed harvester likely resulted from the inherent axial vibration of the magnetostrictive rod. Furthermore, the maximum output energy obtained in this work was 1592 µJ, which was generated by a harvester with 3000 coil turns and two magnets under a load resistance of 3 kΩ. It was found that the maximum output energy was generated when the load resistance was close to the coil impedance at the inherent frequency of the axial vibration of the rod.

This paper presents a simple method of droplet formation using liquids that easily wet polydimethylsiloxane (PDMS) surfaces without any surface treatment. Using only structural features and uniform flow focusing, Oil-in-Water (O/W) and Water-in-Oil-in-Water (W/O/W) droplets were formed in the full PDMS structure. Extrusion channel and three-dimensional flow focusing resulted in effective fluidic conditions for droplet formation and the droplet size could be precisely controlled by controlling the flow rate of each phase. The proposed structure can be utilized as an important element for droplet based research, as well as a droplet generator.

This paper reports design optimization of vibration based energy harvest device fabricated by simple printing and laminating process using polymeric piezoelectric material (VDF/TrFE). Modified trapezoidal design increased the output voltage and energy dramatically by adhesion strength as well as improvement of internal stress distribution of the device. The process guaranteed simple (fast and low cost) fabrication of energy harvest device and the new design achieved 277 V and 1014 nJ outputs which is about 3 times higher than that of our previous device. Furthermore, by stacking the devices and connecting them in parallel, we increased the output energy into about $\mathbf{2.58}\ \mu \mathbf{J}$.

This paper presents methods for the formation of hollow microcapsules and microlenses using multiphase microdroplets. Microdroplets, which consist of a gas core and an organic phase shell, were generated at a single junction on a silicon device without surface treatment of the fluidic channels. Droplet, core and shell dimensions were controlled by varying the flow rates of each phase. When the organic solvent was released from the organic phase shell, the environmental conditions changed the shape of the solidified polymer shell to either a hollow capsule or a microlens. A uniform solvent release process produced polymeric capsules with nanoliter gas core volumes and a membrane thickness of approximately 3 μm. Alternatively physical rearrangement of the core and shell allowed for the formation of polymeric microlenses. On-demand formation of the polymer lenses in wells and through-holes polydimethylsiloxane (PDMS) structures was achieved. Optical properties of the lenses were controlled by changing the dimension of these structures.

This paper proposes a high-throughput, function-based screening approach of a metagenomic library for isolating novel microbial enzymes by droplet-based microfluidics. We used gel microdroplets (GMDs) dispersed in oil as picoliter-volume reaction vessels for lipolytic enzyme by encapsulating cells in individual GMDs. Using this approach, we monitored the growth of individual cells encapsulated in GMDs and assessed the enzyme reaction activities at the level of an individual GMD. We then applied this method to screen lipolytic enzyme genes from the metagenomic library constructed from soil collected from a quercus serrate forest of Mount Tsukuba, Ibaraki, Japan. In the workflow presented in this study, metagenomic library clones were encapsulated in 100-pL GMDs with a fluorogenic reporter substrate. A total of 67,000 metagenomic library clones can be screened in only 24 h with reduced consumption of reagents (i.e.,10 μL). As a result, we identified a novel lipolytic enzyme, EstT1, belonging to the EstD2 family of esterases and containing a putative signal peptide, which facilitates enzyme export and catalyzation of substrates in the periplasm. Our study demonstrates the potential of microfluidic GMDs as an efficient tool for metagenomic library screening of industrially relevant enzymes with the potential of significantly reducing the cost and time factors involved in successful practical application of microbial enzymes.

This paper presents a droplet sampling device driven by horizontal pneumatic actuators. A high aspect ratio and highly flexible PDMS (polydimethylsiloxane) structure was proposed for carrying out larger number of sampling than the number of actuators. Large deformation of the actuators caused domino-deformation of parallel walls, and the deformations allowed for selective collection of target droplets. The dimensions of the PDMS structure and the ratio of resin to curing agent were optimized for efficient sampling under the low pressure applied to the actuators. Five sampling modes were achieved in the simple one-layer structure consisting of one inlet, four walls, one drain channel, and two pneumatic actuators, formed by single-step soft lithography process.

We report a real time method to monitor the chemical reaction in microdroplets, which contain an organic dye, 5(6)carboxynaphthofluorescein and a CdSe/ZnS quantum dot using fluorescence spectra. Especially, the relationship between the droplet diameter and the reaction rate of the two reagents was investigated by changing an inlet flow rate. key words: Microfluidic Device, Fluorescence Spectroscopy, Real-Time Monitoring, Semiconductor Quantum Dot, Fluorescence Energy Transfer

In synthetic biology, the control of gene expression requires a multistep processing of biological signals. The key steps are sensing the environment, computing information and outputting products1. To achieve such functions, the laborious, combinational networking of enzymes and substrate-genes is required, and to resolve problems, sophisticated design automation tools have been introduced2. However, the complexity of genetic circuits remains low because it is difficult to completely avoid crosstalk between the circuits. Here, we have made an orthogonal self-contained device by integrating an actuator and sensors onto a DNA origami-based nanochip that contains an enzyme, T7 RNA polymerase (RNAP) and multiple target-gene substrates. This gene nanochip orthogonally transcribes its own genes, and the nano-layout ability of DNA origami allows us to rationally design gene expression levels by controlling the intermolecular distances between the enzyme and the target genes. We further integrated reprogrammable logic gates so that the nanochip responds to water-in-oil droplets and computes their small RNA (miRNA) profiles, which demonstrates that the nanochip can function as a gene logic-chip. Our approach to component integration on a nanochip may provide a basis for large-scale, integrated genetic circuits.

We report a novel real-time method to monitor the reaction of an organic dye, 5(6)-carboxynaphthofluorescein succinimidyl ester and a CdSe/ZnS quantum dot (QD) using fluorescence spectra of microdroplets formed in a microfluidic device. Dual fluorescence peaks (at λ = 605 and 670 nm) were observed via a fluorescence energy transfer (FRET) process from the CdSe/ZnS QD to the organic dye; the rate of the chemical reaction of the two reagents was estimated by the ratio of the two fluorescence peaks, defined as the fluorescence intensity ratio. In addition, well-controlled microdroplets were continuously generated (with a diameter of ∼150 μm) in the microfluidic device and the reaction time was determined as the length from the mixing point to the sampling point using fluorescence spectroscopy. The fluorescence intensity ratio increased with increasing reaction time after mixing of the two reagents indicating that the chemical reaction was observed directly in the microdroplet. Therefore, the real-time monitoring of the chemical reaction was successfully achieved by measuring the fluorescence intensity ratio as a function of the reaction time. When compared with a conventional batch process, a fast reaction time of

A hydrodynamic droplet pass filter for droplet-phase sample sorting was developed in this study. Using only groove rails, without additional components or external controls, droplets were sorted based on their physical properties. This is the first report of a droplet pass filter used for effective sorting, and the sorting structure provides a novel fluidic component for fluidic circuits for many applications. Depending on the number of rails, we obtained high-pass, low-pass, band-pass, and multi band-pass filters for sorting droplet samples, and their filtration performance was controlled by varying the dimensions of the rail structures. We evaluated in detail the effect of the rail width on sorting, threshold size of droplets sorted into each rail, and capillary number-dependent sample sorting. Furthermore, band-pass droplet sorting, useful for sample quantification, was provided, and multi-step rail ways allowed multi band-pass droplet sorting that was independent of flow conditions. The proposed sorting method does not require any external systems or skilled operation, and thus, it is expected that the device can contribute to on-site sample treatment and analysis in various fields such as medical care or the military.

Depth profiles of absorbed hydrogen introduced by electrochemical charging and light elements were analyzed in Ni-Nb-Zr-H amorphous alloy ribbons using a glow discharge optical emission spectrometer. It was clarified that the absorbed hydrogen was comparatively well-distributed on the sample surface and that the content of the hydrogen decreased with increasing depth from the surface. That is, the amount of absorbed hydrogen on the surface was about 17 at %, while that inside the specimens decreased to several atomic percent. The depth profiles of the hydrogen which were close to the surface were slightly different between those on the roller side and those on the free side in the melt-spun ribbon. The difference is thought to originate from the existence of oxygen impurity on the surface and from the difference of the Zr content.

The effect of the annealing temperature on the magnetostrictive properties of Fe­Co alloy thin films on quartz glass was systematically investigated. The saturation magnetostriction was 56ppm for the Fe32Co68-sputtered thin film quenched from 673K, and it increased to 159ppm with increasing annealing temperature up to 1073K. The magnetostriction above 1093K significantly decreased with the emerging fcc phase in the Fe­Co alloy. SEM images showed that the crystal grains present in the fcc phase aggregated to form a discontinuous surface, which resulted in a decrease in the effective magnetostriction. Measurements of the magnetic properties by vibrating sample magnetometer and magnetic force microscopy revealed the enhancement of the magnetization at the crystal grain boundaries of Fe­Co alloy thin films with large magnetostriction. Thus, the heterogeneity of the magnetization can play a key role in inducing the large magnetostrictive effect. [doi:10.2320/matertrans.MBW201315]

In this study, a fast and quantitative determination method for branched-chain amino acids (BCAAs), namely leucine, isoleucine, and valine, was developed using a pillar array column. A pillar array column with low-dispersion turns was fabricated on a 20 × 20-mm(2) microchip using multistep ultraviolet photolithography and deep reactive ion etching. The BCAAs were fluorescently labeled with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), followed by reversed-phase separation on the pillar array column. The NBD derivatives of the three BCAAs and an internal standard (6-aminocaproic acid) were separated in 100 s. The calibration curves for the NBD-BCAAs had good linearity in the range of 0.4-20 μM, using an internal standard. The intra- and interday precisions were found to be in the ranges of 1.42-3.80 and 2.74-6.97%, respectively. The accuracies for the NBD-BCAA were from 90.2 to 99.1%. The method was used for the analysis of sports drink and human plasma samples. The concentrations of BCAAs determined by the developed method showed good agreements with those determined using a conventional high-performance liquid chromatography system. As BCAAs are important biomarkers of some diseases, these results showed that the developed method could be a potential diagnostic tool in clinical research.

This paper presents a microfluidic system for the active and precise control of microdroplet division in a micro device. Using two horizontal pneumatic valves formed at downstream of bifurcating microchannel, flow resistances of downstream channels were variably controlled. With the resistance control, volumetric ratio of downstream flows was changed and water-in-oil microdroplets were divided into two daughter droplets of different volume corresponding to the ratio. The microfluidic channels and pneumatic valves were fabricated by single-step soft lithography process of PDMS (polydimethylsiloxane) using SU-8 mold. A wide range control of the daughter droplets’ volume ratio was achieved by the simple channel structure. Volumetric ratio between large and small daughter droplets are ranged from 1 to 70, and the smallest droplet volume of 14 pL was obtained. The proposed microfluidic device is applicable for precise and high throughput droplet based digital synthesis.