Cell membrane conformation at vertical nanowire array interface revealed by fluorescence imaging.

The perspectives offered by vertical arrays of nanowires for biosensing applications in living cells depend on the access of individual nanowires to the cell interior. Recent results on electrical access and molecular delivery suggest that direct access is not always obtained. Here, we present a generic approach to directly visualize the membrane conformation of living cells interfaced with nanowire arrays, with single nanowire resolution. The method combines confocal z-stack imaging with an optimized cell membrane labelling strategy which was applied to HEK293 cells interfaced with 2-11 µm long and 3-7 µm spaced nanowires with various surface coatings (bare, aminosilane-coated or polyethyleneimine-coated indium arsenide). We demonstrate that, for all commonly used nanowire lengths, spacings and surface coatings, nanowires generally remain enclosed in a membrane compartment, and are thereby not in direct contact with the cell interior.

Intact mammalian cell function on semiconductor nanowire arrays: new perspectives for cell-based biosensing.

Nanowires (NWs) are attracting more and more interest due to their potential cellular applications, such as delivery of compounds or sensing platforms. Arrays of vertical indium-arsenide (InAs) NWs are interfaced with human embryonic kidney cells and rat embryonic dorsal root ganglion neurons. A selection of critical cell functions and pathways are shown not to be impaired, including cell adhesion, membrane integrity, intracellular enzyme activity, DNA uptake, cytosolic and membrane protein expression, and the neuronal maturation pathway. The results demonstrate the low invasiveness of InAs NW arrays, which, combined with the unique physical properties of InAs, open up their potential for cellular investigations.

Direct observation of acoustic oscillations in InAs nanowires.

Time-resolved X-ray diffraction and optical reflectivity are used to directly measure three different acoustic oscillations of InAs nanowires. The oscillations are excited by a femtosecond laser pulse and evolve at three different time scales. We measure the absolute scale of the initial radial expansion of the fundamental breathing eigenmode and determine the frequency by transient optical reflectivity. For the extensional eigenmode we measure the oscillations of the average radial and axial lattice constants and determine the amplitude of oscillations and the average extension. Finally we observe a bending motion of the nanowires. The frequencies of the eigenmodes are in good agreements with predictions made by continuum elasticity theory and we find no difference in the speed of sound between the wurtzite nanowires and cubic bulk crystals, but the measured strain is influenced by the interaction between different modes. The wurtzite crystal structure of the nanowires however has an anisotropic thermal expansion.

Stages in molecular beam epitaxy growth of GaAs nanowires studied by x-ray diffraction.

GaAs nanowires were grown by molecular beam epitaxy and studied by glancing-angle x-ray diffraction during five different stages of the growth process. An entire forest of randomly positioned epitaxial nanowires was sampled simultaneously and a large variation in the Au-Ga catalyst was found. Au, AuGa, AuGa(2) and the hexagonal beta phase were all identified in several orientations and in similar amounts. The nanowires are shown to consist of regular zinc blende crystal, its twin and the hexagonal wurtzite. The evolution of the various Au-Ga catalysts and the development in the twin to the wurtzite abundance ratio indicate that the Au catalyst is saturated upon initiation of growth leading to an increased amount of wurtzite structure in the wires. A specular x-ray scan identifies the various Au-Ga alloys, three Au lattice constants and a rough interface between nanowires and catalyst. Reciprocal space maps were obtained around Au Bragg points and show the development of the Au catalyst from a distribution largely oriented with respect to the lattice to a non-uniform distribution with several well-defined lattice constants.

Junctions in axial III-V heterostructure nanowires obtained via an interchange of group III elements.

We present an investigation of the morphology and composition of novel types of axial nanowire heterostructures where Ga(x)In(1-x)As is used as barrier material in InAs nanowires. Using aberration-corrected scanning transmission electron microscopy and energy dispersive X-ray analysis we demonstrate that it is possible to grow junctions by changing the group III elements, and we find that a substantial fraction of Ga can be incorporated in axial InAs/Ga(x)In(1-x)As/InAs, retaining straight nanowire configurations. We explain how the adatoms are transferred to the incorporation site at the growth interface via two different routes, (1) interface diffusion and (2) volume diffusion through the catalyst particle.

Molecular beam epitaxy growth of free-standing plane-parallel InAs nanoplates.

Free-standing nanostructures such as suspended carbon nanotubes, graphene layers, III-V nanorod photonic crystals and three-dimensional structures have recently attracted attention because they could form the basis of devices with unique electronic, optoelectronic and electromechanical characteristics. Here we report the growth by molecular beam epitaxy of free-standing nanoplates of InAs that are close to being atomically plane. The structural and transport properties of these semiconducting nanoplates have been examined with scanning electron microscopy, transmission electron microscopy, X-ray diffraction and low-temperature electron transport measurements. The carrier density of the nanoplates can be reduced to zero by applying a voltage to a nearby gate electrode, creating a new type of suspended quantum well that can be used to explore low-dimensional electron transport. The electronic and optical properties of such systems also make them potentially attractive for photovoltaic and sensing applications.

Upscaling and automation of electrophysiology: toward high throughput screening in ion channel drug discovery.

Effective screening of large compound libraries in ion channel drug discovery requires the development of new electrophysiological techniques with substantially increased throughputs compared to the conventional patch clamp technique. Sophion Bioscience is aiming to meet this challenge by developing two lines of automated patch clamp products, a traditional pipette-based system called Apatchi-1, and a silicon chip-based system QPatch. The degree of automation spans from semi-automation (Apatchi-1) where a trained technician interacts with the system in a limited way, to a complete automation (QPatch 96) where the system works continuously and unattended until screening of a full compound library is completed. The performance of the systems range from medium to high throughputs.

Characterization of potassium channel modulators with QPatch automated patch-clamp technology: system characteristics and performance.

Planar silicon chips with 1-2-microm etched holes (average resistance: 2.04 +/- 0.02 MOmega in physiological buffer, n = 274) have been developed for patch-clamp recordings of whole-cell currents from cells in suspension. An automated 16-channel parallel screening system, QPatch 16, has been developed using this technology. A single-channel prototype of the QPatch system was used for validation of the patch-clamp chip technology. We present here data on the quality of patch-clamp recordings and from actual drug screening studies of human potassium channels expressed in cultured cell lines. Using Chinese hamster ovary (CHO) and human embryonic kidney cells (HEK), gigaseals of 4.1 +/- 0.4 GOmega (n = 146) and high-quality whole-cell current recordings were obtained from hERG and KCNQ4 potassium channels. Success rates for gigaseal recordings varied from 40 to 95%, and 67% of the whole-cell configurations lasted for >20 min. Cells were maintained in suspension up to 4 h in a cell storage facility that is integrated in the QPatch 16. No decline in patchability was observed during this time course. A series of screens was conducted with known inhibitors of the hERG and KCNQ4 potassium channels. Dose-response relationship characterizations of verapamil and rBeKm-1 blockage of hERG currents provided IC(50) values similar to values reported in the literature.