We describe the fabrication of elliptical Au nanodisk arrays as a localized surface plasmon resonance (LSPR) sensing substrate for clinical immunoassay via thermal nanoimprint lithography (NIL) and enhancement in the sensitivity of the detection of the prostate-specific antigen (PSA) using the precipitation of 5-bromo-4-chloro-3-indolyl phosphate p-toluidine/nitro blue tetrazolium (BCIP/NBT), catalyzed by alkaline phosphatase. Au nanodisks were fabricated on glass through an unconventional tilted evaporation, which could preserve the thickness of imprinted resists and create an undercut beneficial to the subsequent lift-off process without any damage to pattern dimension and the glass while removing the residual polymers. To investigate the optically anisotropic property of the LSPR sensors, a probe light with linear polarization parallel to and perpendicular to the long axis of the elliptical nanodisk array was utilized, and their sensitivity to the bulk refractive index (RI) was measured as 327 and 167 nm/RIU, respectively. To our knowledge, this is the first application of enzyme-substrate reaction to sandwich immunoassay-based LSPR biosensors that previously suffered from a low sensitivity due to the short penetration depth of the plasmon field, especially when large-sized antibodies were used as bioreceptors. As a result, a large change in local refractive index because of the precipitation on the Au nanodisks amplified the wavelength shift of the LSPR peak in the vis-NIR spectrum, resulting in femtomolar detection limits, which was ∼10(5)-fold lower than the label-free detection without the enzyme precipitation. This method can be extended easily to the other clinical diagnostics with a high sensitivity.

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Nano-imprint lithography, which has the advantages of simplicity, low cost, high replication fidelity and relatively high throughput, requires surface contact between a template with nano patterns and a wafer that transfers the patterns. This article presents the wafer stage for single-step nano-imprint lithography. The wafer stage has a six degree-of-freedom compliant mechanism for complete contact between the surface of the template and the surface of the wafer. The compliant mechanism consists of an inner mechanism for in-plane motion and an outer mechanism for out-of-plane motion. The inner and outer mechanisms have symmetric flexures, which were machined monolithically, onto each plane to cope with thermal deformation. The wafer stage, which was designed to satisfy stiffness requirements, is analyzed with the aid of a dynamic model for a flexure mechanism and a finite element method. Experiments were conducted on the wafer stage in a nano-imprint machine, and nano patterns with linewidths of 100 and 86nm were transferred successfully. This result verifies that the proposed wafer stage can be used in nano-imprint lithography.

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Highly sensitive and reproducible surface enhanced Raman spectroscopy (SERS) requires not only a nanometer-level structural control, but also superb uniformity across the SERS substrate for practical imaging and sensing applications. However, in the past, increased reproducibility of the SERS signal was incompatible with increased SERS sensitivity. This work presents multiple silver nanocrystals inside periodically arrayed gold nanobowls (SGBs) via an electrochemical reaction at an overpotential of -3.0 V (vs. Ag/AgCl). The gaps between the silver nanocrystals serve as hot spots for SERS enhancement, and the evenly distributed gold nanobowls lead to a high device-to-device signal uniformity. The SGBs on the large sample surface exhibit an excellent SERS enhancement factor of up to 4.80 × 10, with excellent signal uniformity (RSD < 8.0 ± 2.5%). Furthermore, the SGBs can detect specific microRNA (miR-34a), which plays a widely acknowledged role as biomarkers in diagnosis and treatment of diseases. Although the small size and low abundance of miR-34a in total RNA samples hinder their detection, by utilizing the advantages of SGBs in SERS sensing, reliable and direct detection of human gastric cancer cells has been successfully accomplished.

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A method to directly collect negatively charged nucleic acids, such as DNA and RNA, in the biosamples simply by applying an electric field in between the sample and collection buffer separated by the nanofilter membrane is proposed. The nanofilter membrane was made of low-stress silicon nitride with a thickness of 100 nm, and multiple pores were perforated in a highly arranged pattern using nanoimprint technology with a pore size of 200 nm and a pore density of 7.22 × 10 8 /cm 2 . The electrophoretic transport of hsa-mir-93-5p across the membrane was confirmed in pure microRNA (miRNA) mimic solution using quantitative reverse transcription-polymerase chain reactions (qRT-PCR). Consistency of the collected miRNA quantity, stability of the system during the experiment, and yield and purity of the prepared sample were discussed in detail to validate the effectiveness of the electrical protocol. Finally, in order to check the applicability of this method to clinical samples, liquid biopsy process was demonstrated by evaluating the miRNA levels in sera of hepatocellular carcinoma patients and healthy controls. This efficient system proposed a simple, physical idea in preparation of nucleic acid from biosamples, and demonstrated its compatibility to biological downstream applications such as qRT-PCR as the conventional nucleic acid extraction protocols.

Experimental and numerical analyses on recovery of polymer deformation after demolding in the hot embossing process J.Thermal imprint lithography or hot embossing is a processing technique using molding to produce surface patterns in polymer resist at micro-and nanoscales. While fast molding is important to improve the yield of the process, the process step that determines the success of imprinting high aspect ratio structures is demolding, a process to separate the mold insert from the patterned resist after conformal molding. In this paper the authors studied the stress and deformation behavior in polymer resist during the cooling and demolding process of thermal imprint lithography via finite element method. A simple model structure of the Si stamp/poly͑methyl methacrylate͒ ͑PMMA͒ resist/Si substrate was used for the simulation, assuming that PMMA is viscoelastic. As demolding proceeds, Von Mises stress in the PMMA layer is highly localized in two locations, one at the transition corner zone between the residual layer and the replicated PMMA pattern and the other close to the contact region with the moving stamp edge, creating two maximums. The presence of the second maximum stress indicates that a structural failure may occur not only when demolding starts, but also immediately before demolding ends. The second maximum stress becomes significant as the angular offset from the ideal normal demolding to the substrate surface increases or for the structures located far away from the symmetric centerline. In addition, the authors will discuss the influence of other parameters, including demolding rate and stamp geometries.