573 research outputs found
Characterization of the Coupling between OutāofāPlane Graphene and Electrogenic Cells
AbstractThe cellāchip coupling is in general regulated by the interplay between cells and the material surface at the interface. Electroactive planar materials have shown limited crosstalk with cells, whereas pseudo 3D patterned materials promote a more intimate contact with the biological system. Here, unprecedented physical properties of a carbonābased material, i.e., graphene, to engineer outāofāplane morphologies are exploited: 1) 3D singleā to fewālayer fuzzy graphene morphology (3DFG), 2) 3DFG on a collapsed Si nanowire mesh template, and 3) 3DFG on a noncollapsed Si nanowire mesh template. These materials are synthesized and interfaced with cardiomyocyteālike cells focusing on the characterization of the cytoskeletal arrangement as well as membrane wrapping processes yet regulated by endocytic proteins. Finally, some major conditions to promote tight coupling to the device and eventually spontaneous intracellular penetration are found
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Synthetically Encoded Ultrashort-Channel Nanowire Transistors for Fast, Pointlike Cellular Signal Detection
Nanostructures, which have sizes comparable to biological functional units involved in cellular communication, offer the potential for enhanced sensitivity and spatial resolution compared to planar metal and semiconductor structures. Silicon nanowire (SiNW) field-effect transistors (FETs) have been used as a platform for biomolecular sensors, which maintain excellent signal-to-noise ratios while operating on lengths scales that enable efficient extra- and intracellular integration with living cells. Although the NWs are tens of nanometers in diameter, the active region of the NW FET devices typically spans micrometers, limiting both the length and time scales of detection achievable with these nanodevices. Here, we report a new synthetic method that combines gold-nanocluster-catalyzed vaporāliquidāsolid (VLS) and vaporāsolidāsolid (VSS) NW growth modes to produce synthetically encoded NW devices with ultrasharp (<5 nm) n-type highly doped to lightly doped (n) transitions along the NW growth direction, where regions serve as source/drain (S/D) electrodes and the n-region functions as an active FET channel. Using this method, we synthesized short-channel SiNW FET devices with independently controllable diameters and channel lengths. SiNW devices with channel lengths of 50, 80, and 150 nm interfaced with spontaneously beating cardiomyocytes exhibited well-defined extracellular field potential signals with signal-to-noise values of ca. 4 independent of device size. Significantly, these āpointlikeā devices yield peak widths of , which is comparable to the reported time constant for individual sodium ion channels. Multiple FET devices with device separations smaller than were also encoded on single SiNWs, thus enabling multiplexed recording from single cells and cell networks with device-to-device time resolution on the order of a few microseconds. These short-channel SiNW FET devices provide a new opportunity to create nanoscale biomolecular sensors that operate on the length and time scales previously inaccessible by other techniques but necessary to investigate fundamental, subcellular biological processes.Chemistry and Chemical BiologyEngineering and Applied Science
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Outside Looking In: Nanotube Transistor Intracellular Sensors
Nanowire-based field-effect transistors, including devices with planar and three-dimensional configurations, are being actively explored as detectors for extra- and intracellular recording due to their small size and high sensitivities. Here we report the synthesis, fabrication, and characterization of a new needle-shaped nanoprobe based on an active silicon nanotube transistor, ANTT, that enables high-resolution intracellular recording. In the ANTT probe, the source/drain contacts to the silicon nanotube are fabricated on one end, passivated from external solution, and then time-dependent changes in potential can be recorded from the opposite nanotube end via the solution filling the tube. Measurements of conductance versus water-gate potential in aqueous solution show that the ANTT probe is selectively gated by potential changes within the nanotube, thus demonstrating the basic operating principle of the ANTT device. Studies interfacing the ANTT probe with spontaneously beating cardiomyocytes yielded stable intracellular action potentials similar to those reported by other electrophysiological techniques. In addition, the straightforward fabrication of ANTT devices was exploited to prepare multiple ANTT structures at the end of single probes, which enabled multiplexed recording of intracellular action potentials from single cells and multiplexed arrays of single ANTT device probes. These studies open up unique opportunities for multisite recordings from individual cells through cellular networks.Chemistry and Chemical Biolog
Intracellular Recordings of Action Potentials by an Extracellular Nanoscale Field-Effect Transistor
The ability to make electrical measurements inside cells has led to many important advances in electrophysiology. The patch clamp technique, in which a glass micropipette filled with electrolyte is inserted into a cell, offers both high signal-to-noise ratio and temporal resolution. Ideally, the micropipette should be as small as possible to increase the spatial resolution and reduce the invasiveness of the measurement, but the overall performance of the technique depends on the impedance of the interface between the micropipette and the cell interior, which limits how small the micropipette can be. Techniques that involve inserting metal or carbon microelectrodes into cells are subject to similar constraints. Field-effect transistors (FETs) can also record electric potentials inside cells, and because their performance does not depend on impedance, they can be made much smaller than micropipettes and microelectrodes. Moreover, FET arrays are better suited for multiplexed measurements. Previously, we have demonstrated FET-based intracellular recording with kinked nanowire structures, but the kink configuration and device design places limits on the probe size and the potential for multiplexing. Here, we report a new approach in which a nanotube is synthetically integrated on top of a nanoscale FET. This nanotube penetrates the cell membrane, bringing the cell cytosol into contact with the FET, which is then able to record the intracellular transmembrane potential. Simulations show that the bandwidth of this branched intracellular nanotube FET (BIT-FET) is high enough for it to record fast action potentials even when the nanotube diameter is decreased to 3Ā nm, a length scale well below that accessible with other methods. Studies of cardiomyocyte cells demonstrate that when phospholipid-modified BIT-FETs are brought close to cells, the nanotubes can spontaneously penetrate the cell membrane to allow the full-amplitude intracellular action potential to be recorded, thus showing that a stable and tight seal forms between the nanotube and cell membrane. We also show that multiple BIT-FETs can record multiplexed intracellular signals from both single cells and networks of cells.Chemistry and Chemical BiologyEngineering and Applied SciencesPhysic
Comparative characterization of the PvuRts1I family of restriction enzymes and their application in mapping genomic 5-hydroxymethylcytosine
PvuRts1I is a modification-dependent restriction endonuclease that recognizes 5-hydroxymethylcytosine (5hmC) as well as 5-glucosylhydroxymethylcytosine (5ghmC) in double-stranded DNA. Using PvuRts1I as the founding member, we define a family of homologous proteins with similar DNA modification-dependent recognition properties. At the sequence level, these proteins share a few uniquely conserved features. We show that these enzymes introduce a double-stranded cleavage at the 3ā²-side away from the recognized modified cytosine. The distances between the cleavage sites and the modified cytosine are fixed within a narrow range, with the majority being 11ā13ānt away in the top strand and 9ā10ānt away in the bottom strand. The recognition sites of these enzymes generally require two cytosines on opposite strand around the cleavage sites, i.e. 5ā²-CN11ā13āN9ā10G-3ā²/3ā²-GN9ā10āN11ā13C-5ā², with at least one cytosine being modified for efficient cleavage. As one potential application for these enzymes is to provide useful tools for selectively mapping 5hmC sites, we have compared the relative selectivity of a few PvuRts1I family members towards different forms of modified cytosines. Our results show that the inherently different relative selectivity towards modified cytosines can have practical implications for their application. By using AbaSDFI, a PvuRts1I homolog with the highest relative selectivity towards 5ghmC, to analyze rat brain DNA, we show it is feasible to map genomic 5hmC sites close to base resolution. Our study offers unique tools for determining more accurate hydroxymethylomes in mammalian cells
Testing sleep consolidation in skill learning: a field study using an online game
Using an observational sample of players of a simple online game (n > 1.2 million), we are
able to trace the development of skill in that game. Information on playing time, and player
location, allows us to estimate time of day during which practice took place. We compare those
whose breaks in practice probably contained a nightās sleep and those whose breaks in practice
probably did not contain a nightās sleep. Our analysis confirms experimental evidence showing
a benefit of spacing for skill learning, but fails to find any additional benefit of sleeping during
a break from practice. We discuss reasons why the well established phenomenon of sleep
consolidation might not manifest in an observational study of skill development. We put the
spacing effect into the context of the other known influences on skill learning: improvement
with practice, and individual differences in initial performance. Analysis of performance data
from games allows experimental results to be demonstrated outside of the lab, and for experimental
phenomenon to be put in the context of the performance of the whole task
Trabecular Meshwork Engineering and Live Tracking in Perfused Porcine Anterior Segments
Purpose: To establish a trabecular meshwork ā¢ engineering model using porcine anterior segments of consistently high quality in a physiological, fixed perfusion system.\ud
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Discussion: Compared to previously used human donor eyes, this inexpensive porcine anterior segment perfusion model is of sufficient, repeatable high quality to develop strategies to modify genetically, ablate and repopulate the TM. Despite significant anatomic differences, effects of transduction and ablation in the porcine model presented here replicate key aspects of previously explored human, feline and rodent models
Interlayer Registry Determines the Sliding Potential of Layered Metal Dichalcogenides: The case of 2H-MoS2
We provide a simple and intuitive explanation for the interlayer sliding
energy landscape of metal dichalcogenides. Based on the recently introduced
registry index (RI) concept, we define a purely geometrical parameter which
quantifies the degree of interlayer commensurability in the layered phase of
molybdenum disulphide (2HMoS2). A direct relation between the sliding energy
landscape and the corresponding interlayer registry surface of 2H-MoS2 is
discovered thus marking the registry index as a computationally efficient means
for studying the tribology of complex nanoscale material interfaces in the
wearless friction regime.Comment: 13 pages, 7 figure
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