UV-nanoimprint lithography as a tool to develop flexible microfluidic devices for electrochemical detection

Research in microfluidic biosensors has led to dramatic improvements in sensitivities. Very few examples of these devices have been commercially successful, keeping this methodology out of the hands of potential users. In this study, we developed a method to fabricate a flexible microfluidic device containing electrowetting valves and electrochemical transduction. The device was designed to be amenable to a roll-to-roll manufacturing system, allowing a low manufacturing cost. Microchannels with high fidelity were structured on a PET film using UV-NanoImprint Lithography (UV-NIL). The electrodes were inkjet-printed and photonically sintered on second flexible PET film. The film containing electrodes was bonded directly to the channel-containing layer to form sealed fluidic device. Actuation of the multivalve system with food dye in PBS buffer was performed to demonstrate automated fluid delivery. The device was then used to detect Salmonella in a liquid sample

Detection of Escherichia coli in Drinking Water Using T7 Bacteriophage-Conjugated Magnetic Probe

In this study, we demonstrate a bacteriophage (phage)-based magnetic separation scheme for the rapid detection of Escherichia colt (E. coli) in drinking water. T7 phage is a lytic phage with a broad host range specificity for E. coli. Our scheme was as follows: (1) T7 bacteriophage-conjugated magnetic beads were used to capture and separate E. coli BL21 from drinking water; (2) subsequent phage-mediated lysis was used to release endemic beta-galactosidase (beta-gal) from the bound bacterial cells; (3) the release of beta-gal was detected using chlorophenol red-beta-D-galactopyranoside (CRPG), a colorimetric substrate which changes from yellow to red in the presence of beta-gal. Using this strategy, we were able to detect E. coli at a concentration of 1 x 10(4) CFU.mL(-1) within 2.5 h. The specificity of the proposed magnetic probes toward E. coli was demonstrated against a background of competing bacteria. By incorporating a pre-enrichment step in Luria-Bertani (LB) broth supplemented with isopropyl beta-D-thiogalactopyranoside (IPTG), we were able to detect 10 CFU.mL(-1) in drinking water after 6 h of pre-enrichment. The colorimetric change can be determined either by visual observation or with a reader, allowing for a simple, rapid quantification of E. coli in resource-limited settings

Surfaces for Competitive Selective Bacterial Capture from Protein Solutions

Active surfaces that form the basis for bacterial sensors for threat detection, food safety, or certain diagnostic applications rely on bacterial adhesion. However, bacteria capture from complex fluids on the active surfaces can be reduced by the competing adsorption of proteins and other large molecules. Such adsorption can also interfere with device performance. As a result, multiple upstream processing steps are frequently employed to separate macromolecules from any cells, which remain in the buffer. Here, we present an economical approach to capture bacteria, without competitive adsorption by proteins, on engineered surfaces that do not employ biomolecular recognition, antibodies, or other molecules with engineered sequences. The surfaces are based on polyethylene glycol (PEG) brushes that, on their own, repel both proteins and bacteria. These PEG brushes backfill the surface around sparsely adsorbed cationic polymer coils (here, poly-r.-lysine (PLL)). The PLL coils are effectively embedded within the brush and produce locally cationic nanoscale regions that attract negatively charged regions of proteins or cells against the steric background repulsion from the PEG brush. By carefully designing the surfaces to include just enough PLL to capture bacteria, but not enough to capture proteins, we achieve sharp selectivity where S. aureus is captured from albumin- or fibrinogen-containing solutions, but free albumin or fibrinogen molecules are rejected from the surface. Bacterial adhesion on these surfaces is not reduced by competitive protein adsorption, in contrast to performance of more uniformly cationic surfaces. Also, protein adsorption to the bacteria does not interfere with capture, at least for the case of S. aureus, to which fibrinogen binds through a specific receptor

Centimeter-long electron transport in marine sediments via conductive minerals

Centimeter-long electron conduction through marine sediments, in which electrons derived from sulfide in anoxic sediments are transported to oxygen in surficial sediments, may have an important influence on sediment geochemistry. Filamentous bacteria have been proposed to mediate the electron transport, but the filament conductivity could not be verified and other mechanisms are possible. Surprisingly, previous investigations have never actually measured the sediment conductivity or its basic physical properties. Here we report direct measurements that demonstrate centimeter-long electron flow through marine sediments, with conductivities sufficient to account for previously estimated electron fluxes. Conductivity was lost for oxidized sediments, which contrasts with the previously described increase in the conductivity of microbial biofilms upon oxidation. Adding pyrite to the sediments significantly enhanced the conductivity. These results suggest that the role of conductive minerals, which are more commonly found in sediments than centimeter-long microbial filaments, need to be considered when modeling marine sediment biogeochemistry

A facile approach to hydrophilic, reverse zwitterionic, choline phosphate polymers

2-{2-(Methacryloyloxy) ethyldimethylammonium}ethyl n-butyl phosphate (MBP) was synthesized and polymerized by controlled free radical methods to afford a new set of ``reverse zwitterionic'' choline phosphate (CP) substituted polymers. The synthesis overcomes prior limitations associated with related CP-methacrylates, affording facile access to MBP homopolymers and MBP-containing diblock copolymers. Dynamic light scattering revealed the size of polyMBP in water, while cell culture assays showed polyMBP to have excellent biocompatibility and non-immunogenicity. The facile synthesis of MBP monomer, and the corresponding polymers, opens a new platform towards a diverse set of functional, hydrophilic biocompatible polymers

Directly Measuring the Complete Stress-Strain Response of Ultrathin Polymer Films

The inherently fragile nature of ultrathin polymer films presents difficulties to the measurement of their mechanical properties, which are of interest in packaging, electronics, separations, and other manufacturing fields. More fundamentally, the direct measurement of ultrathin film mechanical properties is necessary for understanding changes in intrinsic material properties at reduced size scales, for example, when the film thickness alters the equilibrium configuration of the polymer chains. We introduce a method for ultrathin film tensile testing that stretches a two-dimensionally macroscopic, yet nanoscopically thin, polymer film on the surface of water. For polystyrene films, we observe a precipitous decrease in mechanical properties (Young's modulus, strain at failure, and nominal stress at failure) for film thicknesses down to 15 nm, less than the characteristic size of an individual polymer chain, yielding new insights into the changes in polymer chain entanglements in confined states

Low-Temperature Fabrication of Mesoporous Titanium Dioxide Thin Films with Tunable Refractive Indices for One-Dimensional Photonic Crystals and Sensors on Rigid and Flexible Substrates

Highly transparent mesoporous titanium dioxide (TiO2; anatase), thin films were prepared at room temperature via ultraviolet (UV) irradiation Of hybrid polymer-TiO2 nanoparticle thin films: This approach utilized a UV-curable polymer in conjunction with the photocatalytic activity of TiO2 to form and degrade the organic component of the composite films in one step, producing films with well-controlled porosity and refractive index. By adjustment of the loading of TiO2 nanoparticles in the host polymer, the refractive index was tuned between 1.53 and 1.73. Facile control of these properties and mild processing conditions was leveraged to fabricate robust one-dimensional photonic crystals (Bragg mirrors) consisting entirely of TiO2 on silicon and flexible poly(ethylene terephthalate) substrates. The mesoporous Bragg mirrors were shown to be effective chemical vapor sensors with strong optical responses