Reduced graphene oxide (RGO) has recently gained considerable attention for use in electrochemical biosensing applications due to its outstanding conducting properties and large surface area. This report presents a novel microfluidic chip integrated with an RGO-based electrochemical immunosensor for label-free detection of an influenza virus, H1N1. Three microelectrodes were fabricated on a glass substrate using the photolithographic technique, and the working electrode was functionalized using RGO and monoclonal antibodies specific to the virus. These chips were integrated with polydimethylsiloxane microchannels. Structural and morphological characterizations were performed using X-ray photoelectron spectroscopy and scanning electron microscopy. Electrochemical studies revealed good selectivity and an enhanced detection limit of 0.5 PFU mL−1, where the chronoamperometric current increased linearly with H1N1 virus concentration within the range of 1 to 104 PFU mL−1 (R2 = 0.99). This microfluidic immunosensor can provide a promising platform for effective detection of biomolecules using minute samples.
The size of a cell is a fundamental physiological property and is closely regulated by various environmental and genetic factors. Optical or confocal microscopy can be used to measure the dimensions of adherent cells, and Coulter counter or flow cytometry (forward scattering light intensity) can be used to estimate the volume of single cells in a flow. Although these methods could be used to obtain the mass of single live cells, no method suitable for directly measuring the mass of single adherent cells without detaching them from the surface is currently available. We report the design, fabrication, and testing of 'living cantilever arrays', an approach to measure the mass of single adherent live cells in fluid using silicon cantilever mass sensor. HeLa cells were injected into microfluidic channels with a linear array of functionalized silicon cantilevers and the cells were subsequently captured on the cantilevers with positive dielectrophoresis. The captured cells were then cultured on the cantilevers in a microfluidic environment and the resonant frequencies of the cantilevers were measured. The mass of a single HeLa cell was extracted from the resonance frequency shift of the cantilever and was found to be close to the mass value calculated from the cell density from the literature and the cell volume obtained from confocal microscopy. This approach can provide a new method for mass measurement of a single adherent cell in its physiological condition in a non-invasive manner, as well as optical observations of the same cell. We believe this technology would be very valuable for single cell time-course studies of adherent live cells.
Measurements of airborne viruses via sampling have been critical issues. Most electrostatic samplers have been assessed for bacterial aerosols or micrometer-sized viral particles; however, sampling of submicrometer-sized airborne viruses is necessary, especially because of the high probability of their staying airborne and their deposition in the lower respiratory tract. Here, we present a novel personal electrostatic particle concentrator (EPC) for gentle sampling of submicrometer airborne virus particles. Owing to the enhanced electric field designed in this EPC, the collection efficiencies reached values as high as 99.3-99.8% for 0.05-2 μm diameter polystyrene particles at a flow rate of 1.2 L/min. Submicrometer-sized MS2 and T3 virus particles were also collected in the EPC, and the concentrations relative to their respective initial suspensions were more than 10 times higher than those in the SKC BioSampler. Moreover, the recovery rate of T3 was 982 times higher in the EPC (-2 kV) than in the BioSampler at 12.5 L/min because of the gentle sampling of the EPC. Gentle sampling is desirable because many bioaerosols suffer from significant viability losses during sampling. The influence of ozone generated, applied electrostatic field, and the flow rate on the viability of the viruses will also be discussed.
Many ultraviolet (UV)-based disinfection methods have been developed; however, these methods usually use the recirculating mode or need long irradiation periods due to its low photon energy. Vacuum UV (VUV) was recently found to be a promising light source, despite its ozone generation. In this study, we investigated photocatalysis reactions by VUV with short irradiation times (0.004-0.125s) for simultaneously inactivating airborne MS2 viruses and degrading the generated ozone toward a flow-through air disinfection system with high flow-rates. We developed three effective shapes for the catalyst frame: 2mm and 5mm pleated, and spiral-type Pd-TiO 2 catalysts. The 2mm pleated Pd-TiO 2 /VUV photocatalyst exhibited the highest activity ACCEPTED MANUSCRIPT ACCEPTED MANUSCRIPT 2 for simultaneous MS2 inactivation and ozone degradation, and the catalytic activity was effective regardless of relative humidity. Considering the gas phase and catalyst surface effects, and the natural inactivation of VUV-irradiated but live MS2 viruses, the 2mm pleated PdTiO 2 /VUV and succeeding UV photocatalysis showed more than 90% in the overall inactivation efficiency with residual ozone of 35ppb at an irradiation time of 0.009s (flow-rate: 33 l/min). In contrast, most UV-based purifiers take longer times for disinfection. This system has the potential for an alternative to conventional UV-based air purifiers.
The influenza virus has received extensive attention due to the recent H1N1 pandemics originating from swine. This study reports a label-free, highly sensitive, and selective electrical immunosensor for the detection of influenza virus H1N1 based on dielectrophoretically deposited single-walled carbon nanotubes (SWCNTs). COOH-functionalized SWCNTs were deposited on a self-assembled monolayer of polyelectrolyte polydiallyldimethyl-ammonium chloride (PDDA) between two gold electrodes by dielectrophoretic and electrostatic forces, which resulted in reproducible, uniform, aligned, and aggregation-free SWCNT channels (2-10 μm in length). Avidin was immobilized onto the PDDA-SWCNT channels, and viral antibodies were immobilized using biotin-avidin coupling. The resistance of the channels increased with the binding of the influenza viruses to the antibodies. These immunosensors showed linear behavior as the virus concentration was varied from 1 to 10(4) PFU ml(-1) along with a detection time of 30 min. The immunosensors with a 2 μm channel length detected 1 PFU ml(-1) of the influenza virus accurately (R(2) = 0.99) and selectively from MS2 bacteriophages. These immunosensors have the potential to become an important component of a point-of-care test kit that will enable a rapid clinical diagnosis.
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