Assembling Vertical Nanogap Arrays with Nanoentities for Highly Sensitive Electrical Biosensing.

Nanogap biosensors have emerged as promising platforms for detecting and measuring biochemical substances at low concentrations. Although the nanogap biosensors provide high sensitivity, low limit of detection (LOD), and enhanced signal strength, it requires arduous fabrication processes and costly equipment to obtain micro/nanoelectrodes with extremely narrow gaps in a controlled manner. In this work, we report the novel design and fabrication processes of vertical nanogap structures that can electrically detect and quantify low-concentration biochemical substances. Approximately 40 nm gaps are facilely created by magnetically assembling antibody-coated nanowires onto a nanodisk patterned between a pair of microelectrodes. Analyte molecules tagged with conductive nanoparticles are captured and bound to nanowires and bridge over the nanogaps, which consequently causes an abrupt change in the electrical conductivity between the microelectrodes. Using biotin and streptavidin as model antibodies and analytes, we demonstrated that our nanogap biosensors can effectively measure the protein analytes with the LOD of ∼18 pM. The outcome of this research could inspire the design and fabrication of nanogap devices and nanobiosensors, and it would have a broad impact on the development of microfluidics, biochips, and lab-on-a-chip architectures.

An angle-compensating colorimetric strain sensor with wide working range and its fabrication method.

The visual response is one of the most intuitive principles of sensors. Therefore, emission and change of the colors are widely studied for development of chemical, thermal and mechanical sensors. And it is still a challenging issue to fabricate them with a simple working mechanism, high sensitivity, good reliability, and a cost-effective fabrication process. In this study, we propose a mechanical strain sensor, which has 2D photonic crystal structures in nanoscale on stretchable polydimethylsiloxane (PDMS) substrate. Due to the periodic nanostructures, the surface of the sensor produces structural colors. And when it is stretched, the periodicity of the nanostructures changes, which results in the shift of the colors. Multiple nanostructures with different periodicities are integrated on the sensor in order to extend the working range up to 150% with high sensitivity. In addition, reusable and robust molds, which are fabricated by self-assembly of nanoparticles, are used for multiple replications of sensor substrates. Thus, the fabrication process of this study is believed to be potential for possible industrial manufacturing. This study is expected to contribute to strain sensors in the future for the applications of health care, infrastructure monitoring, soft robotics, and wearable devices.

Influence of growth temperature on dielectric strength of Al2O3 thin films prepared via atomic layer deposition at low temperature.

Thin films grown via atomic layer deposition (ALD) suffer from insufficient growth rate and unreliability for temperature-sensitive electronic substrates. This study aimed to examine the growth characteristics and dielectric strength of ALD Al2O3 films grown at low temperatures (≤ 150 °C) for potential application in flexible electronic devices. The growth rate of the Al2O3 films increased from 0.9 to 1.1 Å/cycle with increasing temperature and saturated at growth temperatures ≥ 150 °C, which is the critical temperature at which a complete oxidation reaction occurred. The dielectric strength was also improved with increasing growth temperature, and the films grown at 150 °C showed a high breakdown field strength (~ 8.3 MV/cm), attributable to the decrease in the carbon impurities and oxygen defects, as confirmed by X-ray photoelectron spectroscopy. Even at low growth temperatures (≤ 150 °C), ALD Al2O3 films showed an overall amorphous structure and extremely smooth surfaces regardless of the growth temperature.

A rapid assay provides on-site quantification of tetrahydrocannabinol in oral fluid.

Tetrahydrocannabinol (THC), the primary psychoactive ingredient of cannabis, impairs cognitive and motor function in a concentration-dependent fashion. Drug testing is commonly performed for employment and law enforcement purposes; however, available tests produce low-sensitive binary results (lateral flow assays) or have long turnaround (gas chromatography­mass spectrometry). To enable on-site THC quantification in minutes, we developed a rapid assay for oral THC analysis called EPOCH (express probe for on-site cannabis inhalation). EPOCH features distinctive sensor design such as a radial membrane and transmission optics, all contained in a compact cartridge. This integrated approach permitted assay completion within 5 min with a detection limit of 0.17 ng/ml THC, which is below the regulatory guideline (1 ng/ml). As a proof of concept for field testing, we applied EPOCH to assess oral fluid samples from cannabis users (n = 43) and controls (n = 43). EPOCH detected oral THC in all specimens from cannabis smokers (median concentration, 478 ng/ml) and THC-infused food consumers. Longitudinal monitoring showed a fast drop in THC concentrations within the first 6 hours of cannabis smoking (half-life, 1.4 hours).

Fabrication of robust and reusable mold with nanostructures and its application to anti-counterfeiting surfaces based on structural colors.

In this study, we report a method to fabricate molds and flexible stamps with 2D photonic crystal structures. This includes self-assembly of polystyrene particles into monolayer, oxygen reactive ion etching, thin film (chromium (Cr)) deposition, and polydimethylsiloxane replication. By tuning the thickness of Cr layer, reusable master molds with nano bumps or nano concaves could be prepared selectively. We showed that the replicated flexible stamps out of these molds exhibited structural colors. Characteristics of the colors depended on viewing angle, brightness of background and light source. And the colors even faded out when the background is white or when the stamp was bent. By using this feature, possible strategies for anti-counterfeiting applications have been suggested in this study. Since the molds are reusable and the fabrication method is simple and cost-effective, this study is expected to contribute to nano devices for industries in future.

Rapid and simple single-chamber nucleic acid detection system prepared through nature-inspired surface engineering.

Background: Nucleic acid (NA)-based diagnostics enable a rapid response to various diseases, but current techniques often require multiple labor-intensive steps, which is a major obstacle to successful translation to a clinical setting. Methods: We report on a surface-engineered single-chamber device for NA extraction and in situ amplification without sample transfer. Our system has two reaction sites: a NA extraction chamber whose surface is patterned with micropillars and a reaction chamber filled with reagents for in situ polymerase-based NA amplification. These two sites are integrated in a single microfluidic device; we applied plastic injection molding for cost-effective, mass-production of the designed device. The micropillars were chemically activated via a nature-inspired silica coating to possess a specific affinity to NA. Results: As a proof-of-concept, a colorimetric pH indicator was coupled to the on-chip analysis of NA for the rapid and convenient detection of pathogens. The NA enrichment efficiency was dependent on the lysate incubation time, as diffusion controls the NA contact with the engineered surface. We could detect down to 1×103 CFU by the naked eye within one hour of the total assay time. Conclusion: We anticipate that the surface engineering technique for NA enrichment could be easily integrated as a part of various types of microfluidic chips for rapid and convenient nucleic acid-based diagnostics.

High-Throughput Synthesis of Liposome Using an Injection-Molded Plastic Micro-Fluidic Device.

For mass production of liposomes, we designed a plastic micro-channel device on the basis of 5 µm of micro-nozzle array forming T-junction with 100 µm depth of micro-channel. A micro-channel unit for synthesizing liposomes consisted of two micro-nozzle arrays for mixing two solutions as well as delivery and recovery channels for supplying solutions and collecting liposome suspension. The number of micro-nozzles was approximately 2400 for a micro-channel unit, and seven units were applied independently on a micro-channel plate. The plastic micro-channel plate was injection-molded for mass production using a micro-channel stamper previously fabricated by UV lithography and nickel electroforming process. A plastic cover plate with seven pairs of inlet and outlet ports was machined by mechanical milling and drilling and was assembled with a micro-channel plate using a holder to form a liposome synthesizing device. Flow and mixing of solutions in the micro-channels were tested using colored water to check the micro-fluidic characteristics of the device. Finally, a L-α-phosphatidylcholine (SOY PC) liposome was synthesized using EtOH solution of SOY PC (95%) and saline (0.85% NaOH solution) to find that the liposomes were around 230 and 260 nm in diameter, depending on the flow rate of the lipid solution.

Analysis of the Binding of Analyte-Receptor in a Micro-Fluidic Channel for a Biosensor based on Brownian Motion.

This study experimentally analyses the binding characteristics of analytes mixed in liquid samples flowing along a micro-channel to the receptor fixed on the wall of the micro-channel to provide design tools and data for a microfluidic-based biosensor. The binding or detection characteristics are analyzed experimentally by counting the number of analytes bound to the receptor, with sample analyte concentration, sample flow rate, and the position of the receptor along the micro-channel length as the main variables. A mathematical model is also proposed to predict the number of analytes transported and bound to the receptor based on a probability density function for Brownian motion. The coefficient in the mathematical model is obtained by using a dimensionless mathematical model and the experimental results. The coefficient remains valid for all different conditions of the sample analyte concentration, flow rate, and the position of the receptor, which implies the possibility of deriving a generalized model. Based on the mathematical model derived from mathematical and experimental analysis on the detection characteristics of the microfluidic-based biosensor depending on previously mentioned variables and the height of the micro-channel, this study suggests a design for a microfluidic-based biosensor by predicting the binding efficiency according to the channel height. The results show the binding efficiency increases as the flow rate decreases and as the receptor is placed closer to the sample-injecting inlet, but is unaffected by sample concentration.

Nano sand filter with functionalized nanoparticles embedded in anodic aluminum oxide templates.

Since the ancient Egyptians had used sand as filter media for water purification, its principle has been inherited through generations and it is still being used now in industries. The sand filter consists of sand literally, and the voids within the sand bed are the pores for filtration. Here we present a filtration principle using nanoparticles, so that the voids between the nanoparticles can be considered as effective pores in nanoscale dimension. Anodic aluminum oxide (AAO) membrane has been used as the working template, and the nanoparticles have been injected and embedded within the pores of the AAO template. Nanoparticles with multiple sizes have been used in order to obtain smaller voids. Moreover, the nanoparticles have been functionalized, or electrically charged, with arginine/phenylalanine (RF) peptide group. In this way, filtration performance for charged particles or molecules, such as methylene blue, has been enhanced. Consequently, this study is expected to provide a new principle for fabrication of nano voids, or nano pores, and for filtration in nanoscale dimension.

Rapid identification of health care-associated infections with an integrated fluorescence anisotropy system.

Health care-associated infections (HAIs) and drug-resistant pathogens have become a major health care issue with millions of reported cases every year. Advanced diagnostics would allow clinicians to more quickly determine the most effective treatment, reduce the nonspecific use of broad-spectrum antimicrobials, and facilitate enrollment in new antibiotic treatments. We present a new integrated system, polarization anisotropy diagnostics (PAD), for rapid detection of HAI pathogens. The PAD uses changes of fluorescence anisotropy when detection probes recognize target bacterial nucleic acids. The technology is inherently robust against environmental noise and economically scalable for parallel measurements. The assay is fast (2 hours) and performed on-site in a single-tube format. When applied to clinical samples obtained from interventional procedures, the PAD determined the overall bacterial burden, differentiated HAI bacterial species, and identified drug resistance and virulence status. The PAD system holds promise as a powerful tool for near-patient, rapid HAI testing.

Molecular Dynamics Simulation of the Effect of Angle Variation on Water Permeability through Hourglass-Shaped Nanopores.

Water transport through aquaporin water channels occurs extensively in cell membranes. Hourglass-shaped (biconical) pores resemble the geometry of these aquaporin channels and therefore attract much research attention. We assumed that hourglass-shaped nanopores are capable of high water permeation like biological aquaporins. In order to prove the assumption, we investigated nanoscale water transport through a model hourglass-shaped pore using molecular dynamics simulations while varying the angle of the conical entrance and the total nanopore length. The results show that a minimal departure from optimized cone angle (e.g., 9° for 30 Å case) significantly increases the osmotic permeability and that there is a non-linear relationship between permeability and the cone angle. The analysis of hydrodynamic resistance proves that the conical entrance helps to reduce the hydrodynamic entrance hindrance. Our numerical and analytical results thus confirm our initial assumption and suggest that fast water transport can be achieved by adjusting the cone angle and length of an hourglass-shaped nanopore.

Automated lipid membrane formation using a polydimethylsiloxane film for ion channel measurements.

A black lipid membrane (BLM) is a powerful platform for studying the electrophysiology of cell membranes as well as transmembrane proteins. However, BLMs have disadvantages in terms of stability, accessibility, and transportability, which preclude their industrial applications. To resolve these issues, frozen membrane precursor (MP) was devised to improve the transportability and storability of BLMs. As described previously, MP is a storable and transportable platform that can be delivered to the point-of-use, where BLMs are automatically formed upon thawing at room temperature. However, MP has an inconsistent thinning-out time, ranging from 30 min to 24 h, as well as a low success rate of BLM formation (~27%), which make it undesirable for practical use. In our study, polydimethylsiloxane (PDMS) was introduced as a replacement for conventionally used Teflon film to control thinning-out time. As such, we used a PDMS thin-film, a porous-structured hydrophobic polymer, and squalene, a high viscosity solvent, to facilitate membrane formation, whereas the absorption rates of solvents were controlled to achieve consistent BLM formation time. We successfully reduced thinning-out time down to <1 h as well as enhanced the success rate of BLM formation to greater than 80%. Moreover, we demonstrated the feasibility of our platform for use in drug screening using gramicidin A and guanidine.

Synthetic biomimetic membranes and their sensor applications.

Synthetic biomimetic membranes provide biological environments to membrane proteins. By exploiting the central roles of biological membranes, it is possible to devise biosensors, drug delivery systems, and nanocontainers using a biomimetic membrane system integrated with functional proteins. Biomimetic membranes can be created with synthetic lipids or block copolymers. These amphiphilic lipids and polymers self-assemble in an aqueous solution either into planar membranes or into vesicles. Using various techniques developed to date, both planar membranes and vesicles can provide versatile and robust platforms for a number of applications. In particular, biomimetic membranes with modified lipids or functional proteins are promising platforms for biosensors. We review recent technologies used to create synthetic biomimetic membranes and their engineered sensors applications.

Controlling mold releasing propensity--the role of surface energy and a multiple chain transfer agent.

As the desired feature size of mold-assisted lithography decreases rapidly efficient demolding process becomes more challenging due to strong adhesion between polymeric resists and fine-featured molds. We synthesized new macromolecular additives and investigated the effects of surface energy and contraction of resist materials on demolding propensity by monitoring the adhesion force between the resist and the applied mold. The resist's surface energy was controlled, as inferred from water contact angle measurements, by chemically modifying its hydroxyl functionality. The resist's degree of volume shrinkage during the photocuring procedure was also controlled by mixing in a newly developed chemical that has a multiple radical chain transfer capability. The adhesion force was proportionally reduced as the surface energy of the resist materials decreased and as the volume shrinkage was reduced. When the volume shrinkage control was applied in conjunction with the low surface energy resist material (LS-30UV), we obtained an optimized condition requiring a minimum force for releasing the mold from the cured resist layer.