Henrik Persson

Fibroblasts Cultured on Nanowires Exhibit Low Motility, Impaired Cell Division, and DNA Damage

Nanowires are commonly used as tools for interfacing living cells, acting as biomolecule-delivery vectors or electrodes. It is generally assumed that the small size of the nanowires ensures a minimal cellular perturbation, yet the effects of nanowires on cell migration and proliferation remain largely unknown. Fibroblast behaviour on vertical nanowire arrays is investigated, and it is shown that cell motility and proliferation rate are reduced on nanowires. Fibroblasts cultured on long nanowires exhibit failed cell division, DNA damage, increased ROS content and respiration. Using focused ion beam milling and scanning electron microscopy, highly curved but intact nuclear membranes are observed, showing no direct contact between the nanowires and the DNA. The nanowires possibly induce cellular stress and high respiration rates, which trigger the formation of ROS, which in turn results in DNA damage. These results are important guidelines to the design and interpretation of experiments involving nanowire-based transfection and electrical characterization of living cells.

Fluorescent Nanowire Heterostructures as a Versatile Tool for Biology Applications

Nanowires are increasingly used in biology, as sensors, as injection devices, and as model systems for toxicity studies. Currently, in situ visualization of nanowires in biological media is done using organic dyes, which are prone to photobleaching, or using microscopy methods which either yield poor resolution or require a sophisticated setup. Here we show that inherently fluorescent nanowire axial heterostructures can be used to localize and identify nanowires in cells and tissue. By synthesizing GaP-GaInP nanowire heterostructures, with nonfluorescent GaP segments and fluorescent GaInP segments, we created a barcode labeling system enabling the distinction of the nanowire morphological and chemical properties using fluorescence microscopy. The GaInP photoluminescence stability, combined with the fact that the nanowires can be coated with different materials while retaining their fluorescence, make these nanowires promising tools for biological and nanotoxicological studies.

Vertical oxide nanotubes connected by subsurface microchannels

AbstractWe describe the fabrication of arrays of oxide nanotubes using a combination of bottom up and top down nanofabrication. The nanotubes are made from epitaxially grown semiconductor nanowires that are covered with an oxide layer using atomic layer deposition. The tips of the oxide-covered nanowires are removed by argon sputtering and the exposed semiconductor core is then selectively etched, leaving a hollow oxide tube. We show that it is possible to create fluidic connections to the nanotubes by a combination of electron beam lithography to precisely define the nanotube positions and controlled wet under-etching. DNA transport is demonstrated in the microchannel. Cells were successfully cultured on the nanotube arrays, demonstrating compatibility with cell-biological applications. Our device opens up the possibility of injecting molecules into cells with both spatial and temporal control.

From immobilized cells to motile cells on a bed-of-nails: effects of vertical nanowire array density on cell behaviour.

The field of vertical nanowire array-based applications in cell biology is growing rapidly and an increasing number of applications are being explored. These applications almost invariably rely on the physical properties of the nanowire arrays, creating a need for a better understanding of how their physical properties affect cell behaviour. Here, we investigate the effects of nanowire density on cell migration, division and morphology for murine fibroblasts. Our results show that few nanowires are sufficient to immobilize cells, while a high nanowire spatial density enables a ”bed-of-nails” regime, where cells reside on top of the nanowires and are fully motile. The presence of nanowires decreases the cell proliferation rate, even in the “bed-of-nails” regime. We show that the cell morphology strongly depends on the nanowire density. Cells cultured on low (0.1 μm−2) and medium (1 μm−2) density substrates exhibit an increased number of multi-nucleated cells and micronuclei. These were not observed in cells cultured on high nanowire density substrates (4 μm−2). The results offer important guidelines to minimize cell-function perturbations on nanowire arrays. Moreover, these findings offer the possibility to tune cell proliferation and migration independently by adjusting the nanowire density, which may have applications in drug testing.

Fluid and highly curved model membranes on vertical nanowire arrays.

Sensing and manipulating living cells using vertical nanowire devices requires a complete understanding of cell behavior on these substrates. Changes in cell function and phenotype are often triggered by events taking place at the plasma membrane, the properties of which are influenced by local curvature. The nanowire topography can therefore be expected to greatly affect the cell membrane, emphasizing the importance of studying membranes on vertical nanowire arrays. Here, we used supported phospholipid bilayers as a model for biomembranes. We demonstrate the formation of fluid supported bilayers on vertical nanowire forests using self-assembly from vesicles in solution. The bilayers were found to follow the contours of the nanowires to form continuous and locally highly curved model membranes. Distinct from standard flat supported lipid bilayers, the high aspect ratio of the nanowires results in a large bilayer surface available for the immobilization and study of biomolecules. We used these bilayers to bind a membrane-anchored protein as well as tethered vesicles on the nanowire substrate. The nanowire-bilayer platform shown here can be expanded from fundamental studies of lipid membranes on controlled curvature substrates to the development of innovative membrane-based nanosensors.

Nanofluidics in hollow nanowires.

We present a novel scheme for producing nanotube membranes using free-standing hollow nanowires, with easily controllable dimensions. GaAs-AlInP core-shell nanowires were grown by metal-organic vapor phase epitaxy and were partially embedded in a polymer film. The GaAs core and substrate were etched selectively, leaving tubes with open access to both sides of the membrane. Electrophoretic transport of T4-phage DNA through the hollow nanowires was demonstrated using epifluorescence microscopy.

Enhanced laminin adsorption on nanowires compared to flat surfaces.

Semiconductor nanowires are widely used to interface living cells, and numerous nanowire-based devices have been developed to manipulate or sense cell behavior. We have, however, little knowledge on the nature of the cell-nanowire interface. Laminin is an extracellular matrix protein promoting cell attachment and growth. Here, we used a method based on fluorescence microscopy and measured the relative amount of laminin adsorbed on nanowires compared to flat surfaces. The amount of adsorbed laminin per surface area is up to 4 times higher on 55nm diameter gallium phosphide nanowires compared to the flat gallium phosphide surface between the nanowires. We show that this enhanced adsorption on nanowires cannot be attributed to electrostatic effects, nor to differences in surface chemistry, but possibly to pure geometrical effects, as increasing the nanowire diameter results in a decreased amount of adsorbed protein. The increased adsorption of laminin on nanowires may explain the exceptionally beneficial properties of nanowire substrates for cellular growth reported in the literature since laminin is often used as surface coating prior to cell cultures in order to promote cell growth, and also because primary cell suspensions contain endogenous laminin.

Effect of depth and substrate on use of stream pools by brown trout

Abstract We tested how depth and cobble clusters affected pool use by brown trout Salmo trutta in four enclosures in a seminatural, outdoor stream channel. Into each enclosure, which consisted of a shallow, cobble-filled habitat and a deeper pool, we stocked eight brown trout 9–19 cm long that had been tagged with passive integrated transponders (PIT). Receiver antennae for the PIT tags were placed between the two habitats to allow continuous monitoring of movements of individual fish. We found that trout used pools to a greater extent at night than during the day. Alterations of depth and substrate had no effect on pool use at night. During the day, increasing water depth either alone or together with adding cobbles increased pool use, whereas cobble addition alone had no effect on pool use. We suggest that evaluations of physical habitat modifications in streams should include measurements of population responses as well as information on how, when, and which fish use newly created habitat structures.

Maintaining QoS by utilizing hierarchical wireless systems

Future wireless networks, the systems beyond 3G, will be a combination of different wireless systems. With only one terminal a user should be able to connect to different type of operators, technologies and systems. The issue is also to make the terminal staying online connected, when these changes are made. Both intrasystem handovers and intersystem handovers should be supported by such terminals. The systems will form a hierarchical cell structure, that offer the users several technologies and different type of QoS-levels and bandwidths. To analyze this type of hierarchical cell structure a model has been created, and to verify the model calculations on a Markov-chain have been made. Using the transition probabilities we have calculated the state probabilities, to verify the results from the simulations made of our model. The analytical model and the simulation shows a very good agreement. The results show that the model could be used to show how the WLAN usage will behave as the number of WLANs and WLAN users increase.

Morphology of living cells cultured on nanowire arrays with varying nanowire densities and diameters

Vertical nanowire arrays are increasingly investigated for their applications in steering cell behavior. The geometry of the array is an important parameter, which influences the morphology and adhesion of cells. Here, we investigate the effects of array geometry on the morphology of MCF7 cancer cells and MCF10A normal-like epithelial cells. Different gallium phosphide nanowire array-geometries were produced by varying the nanowire density and diameter. Our results show that the cell size is smaller on nanowires compared to flat gallium phosphide. The cell area decreases with increasing the nanowire density on the substrate. We observed an effect of the nanowire diameter on MCF10A cells, with a decreased cell area on 40 nm diameter nanowires, compared to 60 and 80 nm diameter nanowires in high-density arrays. The focal adhesion morphology depends on the extent to which cells are contacting the substrate. For low nanowire densities and diameters, cells are lying on the substrate and we observed large focal adhesions at the cell edges. In contrast, for high nanowire densities and diameters, cells are lying on top of the nanowires and we observed point-like focal adhesions distributed over the whole cell. Our results constitute a step towards the ability to fine-tune cell behavior on nanowire arrays.