Sonia Grego

Highly flexible transparent electrodes for organic light-emitting diode-based displays

Multilayer indium-tin-oxide (ITO)–Ag–ITO stacks were evaluated as transparent conductors for flexible organic light-emitting diode (OLED) displays. The ITO–metal–ITO (IMI) samples exhibited significantly reduced sheet resistance over ITO and greater than 80% optical transmission. The IMI films deposited on plastic substrates showed dramatically improved mechanical properties when subjected to bending both as a function of radius of curvature as well as number of cycles to a fixed radius. OLEDs were fabricated on both ITO and IMI anodes, and the devices with IMI anodes showed improved performance at current densities greater than 1mA∕cm2 due to the improved conductivity of the anode.

Reverse genetics with a full-length infectious cDNA of the Middle East respiratory syndrome coronavirus

Significance The identification of a novel, emerging human coronavirus with ∼50% mortality, designated Middle East respiratory syndrome coronavirus (MERS-CoV), emphasizes the importance of the rapid development of reagents that can be used to (i) characterize the replication and pathogenesis of emerging pathogens and (ii) develop therapeutics for treatment. In this report, we describe the development of a cassette-based infectious cDNA clone of MERS-CoV and verify that it functions similarly to the wild-type isolate in terms of replication, protein and RNA expression, and spike attachment protein processing. We also show that the virus replicates preferentially in differentiated primary lung cells. Severe acute respiratory syndrome with high mortality rates (∼50%) is associated with a novel group 2c betacoronavirus designated Middle East respiratory syndrome coronavirus (MERS-CoV). We synthesized a panel of contiguous cDNAs that spanned the entire genome. Following contig assembly into genome-length cDNA, transfected full-length transcripts recovered several recombinant viruses (rMERS-CoV) that contained the expected marker mutations inserted into the component clones. Because the wild-type MERS-CoV contains a tissue culture-adapted T1015N mutation in the S glycoprotein, rMERS-CoV replicated ∼0.5 log less efficiently than wild-type virus. In addition, we ablated expression of the accessory protein ORF5 (rMERS•ORF5) and replaced it with tomato red fluorescent protein (rMERS-RFP) or deleted the entire ORF3, 4, and 5 accessory cluster (rMERS-ΔORF3–5). Recombinant rMERS-CoV, rMERS-CoV•ORF5, and MERS-CoV-RFP replicated to high titers, whereas MERS-ΔORF3–5 showed 1–1.5 logs reduced titer compared with rMERS-CoV. Northern blot analyses confirmed the associated molecular changes in the recombinant viruses, and sequence analysis demonstrated that RFP was expressed from the appropriate consensus sequence AACGAA. We further show dipeptidyl peptidase 4 expression, MERS-CoV replication, and RNA and protein synthesis in human airway epithelial cell cultures, primary lung fibroblasts, primary lung microvascular endothelial cells, and primary alveolar type II pneumocytes, demonstrating a much broader tissue tropism than severe acute respiratory syndrome coronavirus. The availability of a MERS-CoV molecular clone, as well as recombinant viruses expressing indicator proteins, will allow for high-throughput testing of therapeutic compounds and provide a genetic platform for studying gene function and the rational design of live virus vaccines.

A Perspective on Nanowire Photodetectors: Current Status, Future Challenges, and Opportunities

One-dimensional semiconductor nanostructures (nanowires (NWs), nanotubes, nanopillars, nanorods, etc.) based photodetectors (PDs) have been gaining traction in the research community due to their ease of synthesis and unique optical, mechanical, electrical, and thermal properties. Specifically, the physics and technology of NW PDs offer numerous insights and opportunities for nanoscale optoelectronics, photovoltaics, plasmonics, and emerging negative index metamaterials devices. The successful integration of these NW PDs on CMOS-compatible substrates and various low-cost substrates via direct growth and transfer-printing techniques would further enhance and facilitate the adaptation of this technology module in the semiconductor foundries. In this paper, we review the unique advantages of NW-based PDs, current device integration schemes and practical strategies, recent device demonstrations in lateral and vertical process integration with methods to incorporate NWs in PDs via direct growth (nanoepitaxy) methods and transfer-printing methods, and discuss the numerous technical design challenges. In particular, we present an ultrafast surface-illuminated PD with 11.4-ps full-width at half-maximum (FWHM), edge-illuminated novel waveguide PDs, and some novel concepts of light trapping to provide a full-length discussion on the topics of: 1) low-resistance contact and interfaces for NW integration; 2) high-speed design and impedance matching; and 3) CMOS-compatible mass-manufacturable device fabrication. Finally, we offer a brief outlook into the future opportunities of NW PDs for consumer and military application.

Piezoelectric scanning mirrors for endoscopic optical coherence tomography

A novel piezoelectric scanning mirror design for endoscopic optical coherence tomography (OCT) is presented. OCT is an interferometric technique providing microscopic tomographic sectioning of biological samples with mm-range penetration capability in tissue and is suited for integration with endoscopes using fiber-based light delivery to the sample. The piezoelectric scanning mirror was designed to provide wide-range rapid forwarding-looking scanning of the optical beam at the distal end of a compact catheter. The optical scanner provides a large ratio of mirror aperture to device size with rectangular mirror sizes ranging from 600 µm × 840 µm to 840 µm × 1600 µm. Static angular displacements up to ±7° (mechanical angle) were demonstrated and resonance frequencies of hundreds of Hz (and up to 1–2 kHz) were measured, depending on the mirror size. The imaging capability of the piezoelectric scanner was demonstrated using a bench-top spectrometer-based Fourier-domain OCT system.

Development and evaluation of bend‐testing techniques for flexible‐display applications

Abstract— Two different approaches to automated bend testing of flexible substrates for display applications were implemented and characterized: a conventional collapsing radius geometry and a novel technique called the “X-Y-θ” geometry. Indium tin oxide (ITO) coated polymer substrates were used to compare the performance of the two automated systems by in-situ electrical-resistance measurements. Manual bending on fixed-diameter mandrels was used to help interpret the results. The advantages and drawbacks of the two systems for providing information of practical use to flexible display R&D are discussed.

Microtubule Treadmilling in Vitro Investigated by Fluorescence Speckle and Confocal Microscopy

Whether polarized treadmilling is an intrinsic property of microtubules assembled from pure tubulin has been controversial. We have tested this possibility by imaging the polymerization dynamics of individual microtubules in samples assembled to steady-state in vitro from porcine brain tubulin, using a 2% glycerol buffer to reduce dynamic instability. Fluorescence speckled microtubules were bound to the cover-glass surface by kinesin motors, and the assembly dynamics of plus and minus ends were recorded with a spinning-disk confocal fluorescence microscopy system. At steady-state assembly, 19% of the observed microtubules (n = 89) achieved treadmilling in a plus-to-minus direction, 34% in a minus-to-plus direction, 37% grew at both ends, and 10% just shortened. For the population of measured microtubules, the distribution of lengths remained unchanged while a 20% loss of original and 27% gain of new polymer occurred over the 20-min period of observation. The lack of polarity in the observed treadmilling indicates that stochastic differences in dynamic instability between plus and minus ends are responsible for polymer turnover at steady-state assembly, not unidirectional treadmilling. A Monte Carlo simulation of plus and minus end dynamics using measured dynamic instability parameters reproduces our experimental results and the amount of steady-state polymer turnover reported by previous biochemical assays.

High-throughput cardiac safety evaluation and multi-parameter arrhythmia profiling of cardiomyocytes using microelectrode arrays.

Microelectrode arrays (MEAs) recording extracellular field potentials of human-induced pluripotent stem cell-derived cardiomyocytes (hiPS-CM) provide a rich data set for functional assessment of drug response. The aim of this work is the development of a method for a systematic analysis of arrhythmia using MEAs, with emphasis on the development of six parameters accounting for different types of cardiomyocyte signal irregularities. We describe a software approach to carry out such analysis automatically including generation of a heat map that enables quick visualization of arrhythmic liability of compounds. We also implemented signal processing techniques for reliable extraction of the repolarization peak for field potential duration (FPD) measurement even from recordings with low signal to noise ratios. We measured hiPS-CMs on a 48 well MEA system with 5minute recordings at multiple time points (0.5, 1, 2 and 4h) after drug exposure. We evaluated concentration responses for seven compounds with a combination of hERG, QT and clinical proarrhythmia properties: Verapamil, Ranolazine, Flecainide, Amiodarone, Ouabain, Cisapride, and Terfenadine. The predictive utility of MEA parameters as surrogates of these clinical effects were examined. The beat rate and FPD results exhibited good correlations with previous MEA studies in stem cell derived cardiomyocytes and clinical data. The six-parameter arrhythmia assessment exhibited excellent predictive agreement with the known arrhythmogenic potential of the tested compounds, and holds promise as a new method to predict arrhythmic liability.

An optically transparent membrane supports shear stress studies in a three-dimensional microfluidic neurovascular unit model

We report a microfluidic blood-brain barrier model that enables both physiological shear stress and optical transparency throughout the device. Brain endothelial cells grown in an optically transparent membrane-integrated microfluidic device were able to withstand physiological fluid shear stress using a hydrophilized polytetrafluoroethylene nanoporous membrane instead of the more commonly used polyester membrane. A functional three-dimensional microfluidic co-culture model of the neurovascular unit is presented that incorporates astrocytes in a 3D hydrogel and enables physiological shear stress on the membrane-supported endothelial cell layer.

Three-dimensional culture conditions differentially affect astrocyte modulation of brain endothelial barrier function in response to transforming growth factor β1.

Blood-brain barrier (BBB) function is regulated by dynamic interactions among cell types within the neurovascular unit, including astrocytes and endothelial cells. Co-culture models of the BBB typically involve astrocytes seeded on two-dimensional (2D) surfaces, which recent studies indicate cause astrocytes to express a phenotype similar to that of reactive astrocytes in situ. We hypothesized that the culture conditions of astrocytes would differentially affect their ability to modulate BBB function in vitro. Brain endothelial cells were grown alone or in co-culture with astrocytes. Astrocytes were grown either as conventional (2D) monolayers, or in a collagen-based gel which allows them to grow in a three-dimensional (3D) construct. Astrocytes were viable in 3D conditions, and displayed a marked reduction in their expression of glial fibrillary acidic protein (GFAP), suggesting reduced activation. Stimulation of astrocytes with transforming growth factor (TGF)β1 decreased transendothelial electrical resistance (TEER) and reduced expression of claudin-5 in co-cultures, whereas treatment of endothelial cells in the absence of astrocytes was without effect. The effect of TGFβ1 on TEER was significantly more pronounced in endothelial cells cultured with 3D astrocytes compared to 2D astrocytes. These results demonstrate that astrocyte culture conditions differentially affect their ability to modulate brain endothelial barrier function, and suggest a direct relationship between reactive gliosis and BBB permeability. Moreover, these studies demonstrate the potential importance of physiologically relevant culture conditions to in vitro modeling of disease processes that affect the neurovascular unit.

High yield fabrication of multilayer polydimethylsiloxane devices with freestanding micropillar arrays

A versatile method to fabricate a multilayer polydimethylsiloxane (PDMS) device with micropillar arrays within the inner layer is reported. The method includes an inexpensive but repeatable approach for PDMS lamination at high compressive force to achieve high yield of pillar molding and transfer to a temporary carrier. The process also enables micropillar-containing thin films to be used as the inner layer of PDMS devices integrated with polymer membranes. A microfluidic cell culture device was demonstrated which included multiple vertically stacked flow channels and a pillar array serving as a cage for a collagen hydrogel. The functionality of the multilayer device was demonstrated by culturing collagen-embedded fibroblasts under interstitial flow through the three-dimensional scaffold. The fabrication methods described in this paper can find applications in a variety of devices, particularly for organ-on-chip applications.