Droplet interface bilayers (DIBs) are physical lipid bilayers that mimic real membranes in living cells, and they are formed quickly using droplets of water and lipids in oil (Fig. 1A). DIBs allow biomolecular sensing and direct detection of transmembrane proteins or peptides and small molecules such as drugs, anesthetics, or even ions. Cell-free expression systems allow in vitro protein synthesis using actual natural machinery extracted from organisms (Fig. 1B). Previous attempts to combine DIBs with cell-free extracts (CFE) encountered bilayer destabilization due to components in the expression system. This study evaluates incorporation of Promega’s T7 S30 High Yield (HY) Expression system with DIBs to pave the way for future in situ expression of light-activated bacteriorhodopsin (BR) and other complex transmembrane proteins in DIBs. A secondary output includes establishing a method for real-time monitoring and modeling of CF expression reactions using minimal volume. The ability to quantify CF output in such small volumes reduces cost per reaction from $20 to around $0.40, and synthesized protein levels reach tens to hundreds of micrograms per milliliter in less than 1 hour at 37°C.

Zinc oxide surface states can be utilized for ultra-specific detection of biomolecules. The major challenges in using ZnO for bio-sensing are attaining enhanced sensitivity and specificity. In this study, we explore the functionalization of zinc in ZnO through utilizing the thiol bond. The purpose of this study is to demonstrate that the ZnO based sensor is capable of achieving high specificity in presence of competitive surface binding through the thiol bond. The final goal is to design an ultra-specific biosensor to detect low occurring biomolecules. In this study, we have selected cortisol as a stress marker to demonstrate quantification and detection from synthetic sweat. In order to demonstrate ultra-specificity, we have used two competitive thiol based molecules binding to zinc, a linker Dithiobis succinimidyl propionate (DSP) and reducing agent of DSP, Dithiothreitol (DTT). Electrochemical impedance spectroscopy (EIS) is used to quantify the signal obtained through various ratiometric concentrations of DSP and DTT. To validate the EIS study results, inherent fluorescence studies are done by mapping changes in green emission spectrum of ZnO before and after linker functionalization. The optimal combination in terms of highest signal is identified to be of 25mM DTT and 50mM DSP. This is implemented in the experiments performed to calibrate the cortisol concentration in synthetic sweat. This study demonstrates the detection of cortisol antigen in synthetic sweat present within the physiological levels of 8 ng/mL to 140 ng/mL.

Fe-Co-B was identified as a potential candidate for the development of high-frequency sensors and high-frequency actuators. To fabricate high-frequency magnetostrictive resonators, Fe-Co-B magetostrictive thin films were prepared by combining electrochemical deposition and microfabrication processes. It is crucial to obtain thin films with proper microstructure and composition. Results showed that Fe-rich Fe-Co-B thin films exhibited better resonance behavior than those with Co-rich and equiatomic Fe and Co compositions. It is also found that the deposition condition plays an important role on the performance of the films. Fe55Co28B17 thin films were fabricated under the same current density for different times. The films exhibited nanocrystalline structure, circular nodules as surface morphology and good resonance behavior. Fe/Co ratio on surface and cross section slightly decreased with increasing the deposition time. The resonance frequency slightly increased and the Q value was found to decrease with increasing deposition time.

The protein survivin (Sur) has been considered as a potential cancer biomarker due to its involvement in disrupting normal cell cycle by stimulating proliferation and inhibiting cell apoptosis. In this work, we have focused on exploring novel platforms for sensitive monitoring of Sur expression, based on molecular beacons and protein modulation of plasmon-controlled fluorescence. In this framework, we show that Sur can be employed in gating the resonance energy transfer (gRET) between fluorescein isothiocyanate probe dye (FITC) and plasmonic citrate-capped gold nanoparticles (AuNP@Cit). Furthermore, we have designed fluorescent dye-bearing molecular beacons (MBs) targeting nucleotides of the survivin mRNA. The antisense oligonucleotide complementary to the target sequence, inserted in the loop area of the hairpin MB structure, has enabled the fluorescence turn-ON MB switching in the presence of the target, thus signaling the high Sur mRNA levels and enhanced Sur protein expression.

We demonstrate fluidically tuned fluorescence enhancement on the colorimetric substrate with the plasmonic effects induced by the periodic gold nanocup arrays. The fluorescence enhancement by the plasmonic effect has been studied extensively by varying the geometries of nanostructures or the morphology of nanoparticles. In this study, however, the fluorescence enhancement without changing these parameters but simply by varying surrounding media on the colorimetric plasmonic surface is accomplished. The dynamic responses of fluorescence from self-assembled monolayer of dyes on the surface were monitored by flowing various fluids with different refractive indices. The dependence of the radiative decay rate as well as the scattering cross-section on the surrounding dielectric properties results into the selective enhancement of the fluorescence intensity, having a maximum at different surrounding refractive index for different fluorophores with different emission band centers.

We review an optofluidic waveguiding lab-on-a-chip used to sense bioparticles. The sensor uses a liquid filled Anti-Resonant Reflecting Optical Waveguide (ARROW) that is interfaced with standard ridge waveguides. The ridge waveguides are coupled to off-chip lasers and detectors via optical fiber. A perpendicular intersection between the ARROW and a ridge waveguide is especially useful for detecting fluorescently tagged particles. Light coupled into the ridge waveguide can fluorescently excite these particles within a very small volume. Fluorescent signal can then be guided within the ARROW and subsequently off chip to a detector.

We also discuss how such devices are fabricated. Both the ARROW and ridge waveguides are made using alternating thin films of tantalum oxide and silicon dioxide on silicon substrates. These thin films are deposited by either sputtering or plasma enhanced chemical vapor deposition (PECVD). The waveguides are patterned using a combination of standard photolithographic processes, reactive ion etching, and sacrificial etching. Low-loss optical guiding is very dependent on both the waveguide structure and the materials used. The latest processes for maximizing detection sensitivity are reviewed.

We also present results using the optofluidic waveguiding sensor for detecting a variety of different types of particles such as fluorescently labeled nanobeads, viruses, ribosomes, and RNA.

Here we present a novel silicon nanopore planar patch clamp chip for single ion channel screening. We fabricate our devices using a combination of KOH and metal-assisted etching. Electrical characterization shows that the shunt capacitance and access resistance are within the accepted ranges for single channel recordings. In order to test our devices, we cultured and differentiated human neuroblastoma SH-SY5Y cells on chip. We reliably obtained a high resistance seal to the cell membrane and report single ion channel activity recordings.

Synthetic nanoparticles have made significant impact across a broad range of technological applications including optical nanoantennas, ultra-sensitive imaging and sensing, and diagnostics and therapeutics. Natural nanoparticles such as viruses and pollutants are major concerns for human health. High-throughput characterization of nanoparticles in terms of their size and shape is crucial for practical applications of synthetic nanoparticles and highly sensitive pathogen identification. Recently, we have demonstrated Interferometric Reflectance Imaging Sensor (IRIS) with the ability to detect single nanoscale particles [1,2].

In single-particle modality of IRIS (SP-IRIS), the interference of light reflected from the sensor surface is modified by the presence of particles producing a distinct signal that reveals the size of the particle. In our approach, the dielectric layered structure acts as an optical antenna optimizing the elastic scattering characteristics of nanoparticles for sensitive detection and analysis. We have demonstrated identification of virus articles in complex samples for various viruses in multiplexed format. Size discrimination of the imaged nanoparticles (virions) allows differentiation between modified viruses having different genome lengths and facilitates a reduction in the counting of non-specifically bound particles to achieve a limit-of-detection (LOD) of 5x103 pfu/mL for the Ebola and Marburg VSV pseudotypes. We have demonstrated the simultaneous detection of multiple viruses in serum or whole blood as well as in samples contaminated with high levels of bacteria [3]. Single nanoparticle detection with IRIS has shown promising results for protein [4] and DNA arrays with attomolar detection sensitivity.

Rare earth (e.g., Eu, Er, Yb, Tm) doped Y2O3 nanocrystals are promising fluorescent bioimaging agents which can overcome well known problems of currently used organic dyes like photobleaching, phototoxicity, and light scattering. Furthermore, the alternative quantum dots (QDs) composed of heavy metals (e.g., CdSe) possess inherently low biocompatibility due to the heavy metal content. In the present work, monodisperse spherical Y2O3:Eu3+ nanocrystals were successfully synthesized by microwave assisted urea precipitation method followed by thermochemical treatment. This is a green, fast and reproducible synthesis method, which is surfactant and hazardous precursors free. The as prepared particles were non-aggregated, spherical particles with a narrow size distribution. The calcined particles have a polycrystalline structure preserving the monodispersity and the spherical morphology of the as prepared particles. After calcination of Y(OH)CO3:Eu3+ precursors at 900°C for 2 hours, a highly crystalline cubic Y2O3 structure was obtained. The Y2O3:Eu3+ spherical particles showed a strong red emission peak at 613nm due to the 5D0–7F2 forced electric dipole transition of Eu3+ ions under UV excitation (235 nm) as revealed by the photoluminescence analysis (PL). The effect of reaction time on size and photoluminescence properties of calcined particles and also the effect of reaction temperature and pressure on the size and the yield of the precipitation process have been studied. The intense red fluorescent emission, excellent stability and potential low toxicity make these QDs promising for applications in bio-related areas such as fluorescence cell imaging or fluorescence bio labels.

A solid-state nanopore was integrated into an optofluidic sensor chip, liquid-core anti-resonant reflecting optical waveguide (ARROW). The solid-state nanopore worked as a smart gate, which simultaneously provided characteristic electrical signals and controlled the entry of single nanoparticles into the liquid-core channel. The subsequent fluorescence detection further identified the nanoparticles by providing optical signals within a specific wavelength range. In this work, correlated electrical and optical detection of single nanoparticles, H1N1 viruses, and λ-DNA molecules was demonstrated. Different types of particles in a mixture were successfully discriminated. Moreover, the flow velocity in the liquid-core channel was extracted with the help of combined analysis of electrical and optical signals. Enhanced electrical sensitivity using a solid-state nanopore with a thin limiting aperture sculpted by SiO2 deposition was also shown.