We combine nanofluidics and nanoplasmonics for surface-plasmon resonance (SPR) sensing using flow-through nanohole arrays. The role of surface plasmons on resonant transmission motivates the application of nanohole arrays as surface-based biosensors. Research to date, however, has focused on dead-ended holes, and therefore failed to harness the benefits of nanoconfined transport combined with SPR sensing. The flow-through format enables rapid transport of reactants to the active surface inside the nanoholes, with potential for significantly improved time of analysis and biomarker yield through nanohole sieving. We apply the flow-through method to monitor the formation of a monolayer and the immobilization of an ovarian cancer biomarker specific antibody on the sensing surface in real-time. The flow-through method resulted in a 6-fold improvement in response time as compared to the established flow-over method.
Plasmonics is a fast developing research area with a great potential for practical applications. However, the implementation of plasmonic devices requires low cost methodologies for the fabrication of organized metallic nanostructures that covers a relative large area (∼1 cm2). Here the patterning of periodic arrays of nanoholes (PANHs) in gold films by using a combination of interference lithography, metal deposition, and lift off is reported. The setup allows the fabrication of periodic nanostructures with hole diameters ranging from 110 to 1000 nm, for 450 and 1800 nm of periodicity, respectively. The large areas plasmonic substrates consist of 2 cm × 2 cm gold films homogeneously covered by nanoholes and gold films patterned with a regular microarray of 200 μm diameter circular patches of PANHs. The microarray format is used for surface plasmon resonance (SPR) imaging and its potential for applications in multiplex biosensing is demonstrated. The gold films homogeneously covered by nanoholes are useful as electrodes in a thin layer organic photovoltaic. This is first example of a large area plasmonic solar cell with organized nanostructures. The fabrication approach reported here is a good candidate for the industrial‐scale production of metallic substrates for plasmonic applications in photovoltaics and biosensing.
An in-hole nanohole surface plasmon resonance sensing scheme is demonstrated. Arrays of periodic nanoholes milled through thin layers of SiO(x) and gold were used to detect the binding of organic and biological molecules inside the nanoholes, while blocking the gold surfaces outside the holes. This new approach is more efficient than the previous nanohole array method, where the response was related to binding events taking place inside of the holes and on the top gold surface. The improved sensitivity to binding events and lower detection limit are related to resonant surface plasmon enhanced transmission through the arrays of nanoholes. The sensitivity was found to be 650 nm/RIU and the detection of three attomoles of proteins was estimated from this scheme.
The need for early detection of various diseases, including breast cancer, has motivated research into nanomaterials that can be assembled in organized films which serve as biosensors. Owing to the variety of possible materials and film architectures, procedures are required to design optimized biosensors. In this study, we combine surface-specific methods to monitor the assembly of antibodies on nanostructured films with two distinct architectures. In the first, a layer of the antibody type mouse anti-HER2 (clone tab250) was immobilized on a self-assembled monolayer (SAM) of 11-mercaptoundecanoic acid modified with N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide (EDC). In the second approach, a SAM of cysteamine was coated with a biotin/spreptavidin bilayer on which a layer of biotinylated antibody type MSx2HUp185/her biotin was adsorbed. The rougher, less passivating coating with cysteamine determined from cyclic voltammetry and scanning electron microscopy led to biosensors that are more sensitive to detect the breast cancer ERBB2 (HER2) biomarker in impedance spectroscopy measurements. This higher distinguishing ability of the cysteamine-containing film architecture was proven with information visualization methods to treat the impedance data. Polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) confirmed that biosensing resulted from the antibody-ERBB2 antigen affinity.
Nanohole array-based biosensors integrated with a microfluidic concentration gradient generator were used for imaging detection and quantification of ovarian cancer markers. Calibration curves based on controlled concentrations of the analyte were created using a microfluidic stepped diffusive mixing scheme. Quantification of samples with unknown concentration of analyte was achieved by image-intensity comparison with the calibration curves. The biosensors were first used to detect the immobilization of ovarian cancer marker antibodies, and subsequently applied for the quantification of the ovarian cancer marker r-PAX8 (with a limit of detection of about 5 nM and a dynamic range from 0.25 to 9.0 μg.mL(-1)). The proposed biosensor demonstrated the ability of self-generating calibration curves on-chip in an integrated microfluidic platform, representing a further step towards the development of comprehensive lab-on-chip biomedical diagnostics based on nanohole array technology.
Acquired immunodeficiency syndrome HIV/(AIDS) infection has become a chronic disease. This change led to a growing interest in outcomes focusing on the individual, such as health-related quality-of-life (QoL), to assess the impact of treatments. Our study evaluated the impact of the use of antiretroviral drugs, factors related to the infection and socioeconomic factors on the HQoL of HIV-infected patients. A total of 367 patients answered the questionnaire based on the World Health Organization QoL for HIV brief version. According to the modified-Poisson regression approach, the use of antiretroviral drugs was associated with worse QoL in the domain of the level of independence (P = 0.016). Being unemployed was associated with a worse QoL (P < 0.05) in five out of six domains; the only exception was the domain of spirituality (P > 0.05). Among the factors assessed, the socioeconomic aspects were most significant in determining the HQoL of the patients studied.
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