The waveguide-coupled bimetallic (WcBiM) surface plasmon resonance (SPR) chip had been utilized in the intensity interrogation detection mode to detect amyloid-β42 (Aβ42), a biomarker of the Alzheimer disease. The SPR reflectance curve of the WcBiM chip has the narrower full-width-at-half-maximum (FWHM) compared with the SPR reflectance curve of the conventional gold (Au) chip, resulting in the steeper gradient. For the enhancement of resolution, the light source was fixed at an angle where the slope of the reflectance curve is the steepest, and the change in the reflectance was monitored. For the detection of Aβ42, the antibody of Aβ42 (anti-Aβ42) was immobilized on the WcBiM SPR chip using the self-assembled monolayer. The SPR responses, the average changes in the reflectance to the Aβ42 at the concentrations of 100 pg/ml, 250 pg/ml, 500 pg/ml, 750 pg/ml, 1,000 pg/ml, and 2,000 pg/ml were 0.0111%, 0.0305%, 0.0867%, 0.1712%, 0.3021%, and 0.5577%, respectively, for the three replicates. From linear regression analysis, the calibration curve indicated that the SPR response had a linear relation with Aβ42 with the concentration in the range of 100 pg/ml to 2,000 pg/ml. A control experiment showed the anti-Aβ42-modified surface of the WcBiM chip had a high specificity to Aβ42. Thus, the enhanced resolution by utilizing the WcBiM SPR chip in the intensity interrogation detection mode aids the diagnosis of the Alzheimer disease by detecting the Aβ42 around the criteria concentration (500 pg/ml) without any labeling.

Quantitative understanding of the mechanical behavior of biological liquid crystals such as proteins is essential for gaining insight into their biological functions, since some proteins perform notable mechanical functions. Recently, single-molecule experiments have allowed not only the quantitative characterization of the mechanical behavior of proteins such as protein unfolding mechanics, but also the exploration of the free energy landscape for protein folding. In this work, we have reviewed the current state-of-art in single-molecule bioassays that enable quantitative studies on protein unfolding mechanics and/or various molecular interactions. Specifically, single-molecule pulling experiments based on atomic force microscopy (AFM) have been overviewed. In addition, the computational simulations on single-molecule pulling experiments have been reviewed. We have also reviewed the AFM cantilever-based bioassay that provides insight into various molecular interactions. Our review highlights the AFM-based single-molecule bioassay for quantitative characterization of biological liquid crystals such as proteins.

The characteristics of a waveguide-coupled bimetallic (WcBiM) chip in a miniaturized surface plasmon resonance (SPR) sensor and its detection capability for a low molecular weight biomolecule were investigated. The configuration of the WcBiM chip was gold (Au)/waveguide (ZnS-SiO2)/silver (Ag). In the intensity measurement mode, the sensitivity could be improved by reducing the full width at half maximum (FWHM) of the reflectance curve. The FWHM of the WcBiM chip is narrower than that of the Au chip, which suggests that the slope of the reflectance curve for the WcBiM chip is steeper. In order to generate enhanced resolution, the reflectance should be monitored at the specific angle where the slope is the steepest in the reflectance curve. For the detection of biotin that is a low molecular weight biomolecule, streptavidin was formed on the SPR sensor chip surface. The response of the SPR to biotin at various concentrations was then acquired. The sensitivities of the WcBiM chip and the Au chip were 0.0052%/(ng/ml) and 0.0021%/(ng/ml), respectively. The limit of detection of the biotin concentration for both the WcBiM and Au chips was calculated. The values were 2.87 ng/ml for the WcBiM chip and 16.63 ng/ml for the Au chip. Enhancement of the sensitivity in the intensity detection mode was achieved using the WcBiM chip compared with the Au chip. Therefore, sufficient sensitivity for the detection of a disease-related biomarker is attainable with the WcBiM chip in the intensity measurement mode using a miniaturized SPR sensor.

In this study, parylene-C films were deposited using vertically aligned-chemical vapor condensation (CVC) method and effects of post-annealing were investigated. The comparison results for as-deposited and post-annealed samples showed that the structural properties changed gradually with varying annealing temperature, while optical properties and surface morphology of the films seldom changed. The crystallinity, crystallite size, and d-spacing were changed from 60 to 87.2%, from 6.83 to 15.6 nm, and from 6.41 to 6.26 Å, respectively. The value of oxygen transmission rate (OTR) for the films was changed gradually from 3.22 to 1.53 cc/m²•day after annealing process. However, the values of transmittance and surface roughness were 90% in the visible-near IR region and 5 nm without reference to annealing temperature. Consequently, the crystallization of parylene-C films was one of major effects without the change of optical properties and surface morphology after annealing process, and we believed that the enhancement of crystallinity caused to improve the property of OTR for parylene-C films.