Nanoscale-tipped wire array injections transfer DNA directly into brain cells ex vivo and in vivo.

Genetic modification to restore cell functions in the brain can be performed through the delivery of biomolecules in a minimally invasive manner into live neuronal cells within brain tissues. However, conventional nanoscale needles are too short (lengths of ~10 µm) to target neuronal cells in ~1-mm-thick brain tissues because the neuronal cells are located deep within the tissue. Here, we report the use of nanoscale-tipped wire (NTW) arrays with diameters < 100 nm and wire lengths of ~200 µm to address biomolecule delivery issues. The NTW arrays were manufactured by growth of silicon microwire arrays and nanotip formation. This technique uses pinpoint, multiple-cell DNA injections in deep areas of brain tissues, enabling target cells to be marked by fluorescent protein (FP) expression vectors. This technique has potential for use for electrophysiological recordings and biological transfection into neuronal cells. Herein, simply pressing an NTW array delivers and expresses plasmid DNA in multiple-cultured cells and multiple-neuronal cells within a brain slice with reduced cell damage. Additionally, DNA transfection is demonstrated using brain cells ex vivo and in vivo. Moreover, knockdown of a critical clock gene after injecting a short hairpin RNA (shRNA) and a genome-editing vector demonstrates the potential to genetically alter the function of living brain cells, for example, pacemaker cells of the mammalian circadian rhythms. Overall, our NTW array injection technique enables genetic and functional modification of living cells in deep brain tissue areas, both ex vivo and in vivo.

Nanoscale-Tipped High-Aspect-Ratio Vertical Microneedle Electrodes for Intracellular Recordings.

Intracellular recording nanoscale electrode devices provide the advantages of a high spatial resolution and high sensitivity. However, the length of nanowire/nanotube-based nanoelectrodes is currently limited to <10 µm long due to fabrication issues for high-aspect-ratio nanoelectrodes. The concept reported here can address the technological limitations by fabricating >100 µm long nanoscale-tipped electrodes, which show intracellular recording capability.

Nanoscale tipped microwire arrays enhance electrical trap and depth injection of nanoparticles.

Nanoscale devices have the potential to measure biological tissues as well as individual cells/neurons. However, three-dimensional (3D) multi-site probing remains problematic because only planar-type device designs are applicable to sample surfaces. Herein we report 3D nanoscale electrode tipped microwire arrays with high aspect ratios. A nanoscale tipped wire is formed by isotropic silicon etching to the tip of a vapor-liquid-solid grown silicon microwire. After coating the wire with a metal (e.g., Pt and Au), only the nanotip section can be exposed from the surrounding outer shell (e.g., SiO(2) and parylene) by photoresist spray coating and subsequent cycled photoresist etchings. As a promising device application, we demonstrate the trapping of polystyrene nanoparticles in a solution using a fabricated Au-nanotip wire array. The sharpened nanotip has a 150 nm curvature radius and a 4.2 µm(2) electrode area. The nanotip wires exhibit a locally enhanced trapping performance with a low trapping voltage of 20 mV. Moreover, these trapped nanoparticles can be injected into a soft material (gelatin), demonstrating a multi-site wide-area batch depth injection and an assembly of nanoparticles. Such nanotip wire arrays should be applicable to trap numerous particles, including DNA/molecules attached to Au particles, and may realize injection into biological tissues and individual cells/neurons.

Nanoscale sharpening tips of vapor-liquid-solid grown silicon microwire arrays.

We developed out-of-plane, high aspect ratio, nanoscale tip silicon microwire arrays for application to penetrating, multisite, nanoscale biological sensors. Silicon microwire arrays selectively grown by gold-catalyzed vapor-liquid-solid growth of silicon can be formed to create sharpened nanotips with a tip diameter of less than 100 nm by utilizing batch-processed silicon chemical etching for only 1-3 min. The tip angles achieved ranged from 11 degrees to 38 degrees. The nanotip silicon microwires can perform gelatin penetration without wire breakdown, indicating their potential penetrating capability for measurements inside biological tissues.