Rapid ELISA Using a Film-Stack Reaction Field with Micropillar Arrays.

A film-stack reaction field with a micropillar array using a motor stirrer was developed for the high sensitivity and rapid enzyme-linked immunosorbent assay (ELISA) reaction. The effects of the incubation time of a protein (30 s, 5 min, and 10 min) on the fluorescence intensity in ELISAs were investigated using a reaction field with different micropillar array dimensions (5-µm, 10-µm and 50-µm gaps between the micropillars). The difference in fluorescence intensity between the well with the reaction field of 50-µm gap for the incubation time of 30 s and the well without the reaction field with for incubation time of 10 min was 6%. The trend of the fluorescence intensity in the gap between the micro pillars in the film-stack reaction field was different between the short incubation time and the long incubation time. The theoretical analysis of the physical parameters related with the biomolecule transport indicated that the reaction efficiency defined in this study was the dominant factor determining the fluorescence intensity for the short incubation time, whereas the volumetric rate of the circulating flow through the space between films and the specific surface area were the dominant factors for the long incubation time.

Enhancement of Convection and Molecular Transport into Film Stacked Structures by Introduction of Notch Shape for Micro-Immunoassay.

A 3D-stack microfluidic device that can be used in combination with 96-well plates for micro-immunoassay was developed by the authors. ELISA for detecting IgA by the 3D-stack can be performed in one-ninth of the time of the conventional method by using only 96-well plates. In this study, a notched-shape film was designed and utilized for the 3D-stack to promote circulation by enhancing and utilizing the axial flow and circumferential flow in order to further reduce the reaction time. A finite element analysis was performed to evaluate the axial flow and circumferential flow while using the 3D-stack in a well and design the optimal shape. The 3D-stack with the notched-shape film was fabricated and utilized for the binding rate test of the antibody and antigen and ELISA. As a result, by promoting circulation using 3D-stack with notched-shape film, the reaction time for each process of ELISA was reduced to 1 min, which is 1/60 for 96 wells at low concentrations.

Enhancement of Molecular Transport into Film Stacked Structures for Micro-Immunoassay by Unsteady Rotation.

A film-stacked structure consisting of polyethylene terephthalate (PET) films stacked in a gap of 20 µm that can be combined with 96-well microplates used in biochemical analysis has been developed by the authors. When this structure is inserted into a well and rotated, convection flow is generated in the narrow gaps between the films to enhance the chemical/bio reaction between the molecules. However, since the main component of the flow is a swirling flow, only a part of the solution circulates into the gaps, and reaction efficiency is not achieved as designed. In this study, an unsteady rotation is applied to promote the analyte transport into the gaps using the secondary flow generated on the surface of the rotating disk. Finite element analysis is used to evaluate the changes in flow and concentration distribution for each rotation operation and to optimize the rotation conditions. In addition, the molecular binding ratio for each rotation condition is evaluated. It is shown that the unsteady rotation accelerates the binding reaction of proteins in an ELISA (Enzyme Linked Immunosorbent Assay), a type of immunoassay.

Constitutive Model of the Surface Roughening Behavior of Austenitic Stainless Steel.

The martensitic phase transformation (MPT) is one of the most important factors that enhances the surface roughening of stainless-steel thin metal foils (TMF), such as SUS 304, compared to others without MPT, even in the same plastic strain. However, the conventional roughening model does not take into account the influence of MPT. In this study, the authors proposed a new constitutive model to express the surface roughening by taking the influence of MPT into account. The volume fractions of MPT for TMF of SUS304 in various grain sizes are accounted for quantitatively after the tensile test at room temperature and an elevated temperature, and the effect of MPT on the surface roughening is evaluated in comparison to using TMF of SUS316, in which MPT does not occur during plastic deformation. Then, a constitutive model of the surface roughening based on the experimental results is successfully built.

Effect of Peak Current Density on Tensile Properties of AZ31B Magnesium Alloy.

An investigation into the effects, including the athermal effect, of a pulsed current on AZ31B magnesium alloy was carried out. Different peak current densities were applied at the same temperature under uniaxial tensile testing. The results indicate that the stress reduction caused by the increasing peak current density is independent of temperature. The strain hardening coefficient also shows a similar trend. The fracture strain shows the optimum value due to the current crowding effect.

Evolution of acoustic softening effect on ultrasonic-assisted micro/meso-compression behavior and microstructure.

The application of ultrasonic vibration is an effective method to overcome the processing problems in micro/meso-forming. Previously it was observed that ultrasonic vibration could reduce flowing stress in the forming process, called ultrasonic volume effect. The volume effect contains multi-mechanisms such as stress superposition leading to apparent average stress reduction, acoustic softening and ultrasonic impact leading to real stress reductin. However, the evolutional characteristics and the mechanism of acoustic softening on material deformation is still not clear. And in most previous studies only the average stress but not the oscillatory stress was measured due to the convenience of dynamic force sensing system, which confused the different ultrasonic volume effects, acoustic softening, stress superposition and ultrasonic impact. The purpose of this study is to investigate the effects of acoustic softening on micro/meso-compression behavior and microstructure evolution. An ultrasonic-assisted compression test system with dynamic force sensing technology was developed. And a series of ultrasonic-assisted micro/meso-compression tests at different amplitudes were carried out on pure copper C1100O combining the microstructure analysis by EBSD technique. By analyzing the waveform of the oscillatory stress in the process, acoustic softening was successfully separated from the stress superposition and it was found that the deformation strain plays an important role on the effect of acoustic softening. The stress reduction by acoustic softening increases with the flowing strain or ultrasonic amplitude increasing. Besides, there is an evolutionary transition of acoustic softening ratio between small strain and large strain. When acoustic softening occurs, the low-angle grain boundaries distribute randomly in grains, compared to the piled distribution without ultrasonic assistance, implying motions of the low-angle grain boundaries or dislocation is improved by acoustic softening, resulting in the real stress reduction. In addition, with small deformation strain, the elongated grain becomes equiaxed and dislocation density is significantly reduced, which may be the result of the increased dislocation annihilation due to ultrasonic-induced dynamic recovery. However, with the deformation strain increasing to some extent, acoustic hardening gradually becomes significant, leading to much less effectiveness of acoustic softening on dislocation density reduction. The findings of this study provide an instructive understanding of the underlying mechanisms of acoustic softening in ultrasonic-assisted micro/meso-forming.

Investigation on ultrasonic volume effects: Stress superposition, acoustic softening and dynamic impact.

Conventional high power ultrasonic vibration has been widely used to improve manufacturing processes like surface treatment and metal forming. Ultrasonic vibration affects material properties, leading to a flow stress reduction, which is called ultrasonic volume effect. The volume effect contains multi-mechanisms such as stress superposition due to oscillatory stress, acoustic softening by easier dislocation motion and dynamic impact leading to extra surface plastic deformation. However, most researches ignored the stress superposition for the convenience of measurement, and few studies considered ultrasonic dynamic impact since the relatively low ultrasonic energy in macro scale. The purpose of this study is to investigate the characteristics and mechanisms of different ultrasonic volume effects in micro-forming. A 60 kHz longitudinal ultrasonic-assisted compression test system was developed and a series of ultrasonic-assisted compression tests at different amplitudes on commercially pure aluminum A1100 in micro-scale were carried out combining the surface analysis by SEM, EDX and micro-hardness test. Three different ultrasonic volume effects, stress superposition, acoustic softening and dynamic impact, were confirmed in the ultrasonic-assisted compression tests. In order to quantitatively predict stress superposition, a hybrid model for stress superposition is developed considering the elastic deformation of experimental apparatus in practice, the evolution of the modeling results fitted well with the experimental results. With low ultrasonic amplitude, stress superposition and acoustic softening occurred because vibrated punch contacted with the specimen all the time during compression. However, with higher amplitude, due to the extra surface plastic deformation by larger ultrasonic energy, forming stress was further reduced by the ultrasonic dynamic impact. A possible method to distinguish the effects of dynamic impact and acoustic softening is to analyze the waveform of the oscillatory stress in the process. In the case of ultrasonic dynamic impact effect, a higher amount of oxidation was observed on the specimen surface, which could be the result of local heating by surface plastic deformation and surface friction when the vibrated punch detached from the specimen. The findings of this study provide an instructive understanding of the underlying mechanisms of volume effects in ultrasonic-assisted micro-forming.

Computational simulation of biomolecules transport with multi-physics near microchannel surface for development of biomolecules-detection devices.

BACKGROUND: The quantitative evaluation of the biomolecules transport with multi-physics in nano/micro scale is demanded in order to optimize the design of microfluidics device for the biomolecules detection with high detection sensitivity and rapid diagnosis. OBJECTIVE: This paper aimed to investigate the effectivity of the computational simulation using the numerical model of the biomolecules transport with multi-physics near a microchannel surface on the development of biomolecules-detection devices. METHODS: The biomolecules transport with fluid drag force, electric double layer (EDL) force, and van der Waals force was modeled by Newtonian Equation of motion. The model validity was verified in the influence of ion strength and flow velocity on biomolecules distribution near the surface compared with experimental results of previous studies. The influence of acting forces on its distribution near the surface was investigated by the simulation. RESULTS: The trend of its distribution to ion strength and flow velocity was agreement with the experimental result by the combination of all acting forces. Furthermore, EDL force dominantly influenced its distribution near its surface compared with fluid drag force except for the case of high velocity and low ion strength. CONCLUSIONS: The knowledges from the simulation might be useful for the design of biomolecules-detection devices and the simulation can be expected to be applied on its development as the design tool for high detection sensitivity and rapid diagnosis in the future.