University of California

The Penn State Research Foundation

Biotechnology Research and Development Corporation

Other

Harvard College

The Brigham and Women's Hospital, Inc.

Nanyang Technological University

University of South Carolina

Penn State Research Foundation and Biotechnology Research

The Penn State Research Foundation, Penn State Rsrch Foundtn &

Paul S Weiss

Stephan J Stranick

Steven J Jonas

Jason N Belling

Anne M Andrews

Xiaobin Xu

Qing Yang

Natcha Wattanatorn

James Mitchell Tour

Richard Lawrence Baer

Joanna Aizenberg

Mark A Reed

Stanley Joel Osher

Hsian-Rong Tseng

Nam-Joon Cho

Barry G Willis

David Allara

Alireza Khademhosseini

Andrea L Bertozzi

Gregory S McCarty

Alireza Moshaverinia

Jorge M Seminario

Mohammad Mahdi Hasani-Sadrabadi

Tara L Aghaloo

James Nathan Hohman

Xu Hou

Joshua A A Jackman

Gail Vinnacombe

Leonardo Scarabelli

Chuanzhen Zhao

Yao Gong

Liv Heidenreich

Logan A Stewart

Naihao Chiang

Kevin M Cheung

Anat Hatzor

Ray L Funk

Adam Z Stieg

Dan Wilkinson

Lloyd A Bumm

A microfluidic device is disclosed that is used to process cells for the intracellular delivery of molecules or other cargo. The device includes one or more microchannels disposed in a substrate or chip and is fluidically coupled to an inlet configured to receive a solution containing the cells and the molecules or other cargo to be delivered intracellularly to the cells. Each of the one or more microchannels has one or more constriction regions formed therein, wherein the inner surface(s) of the microchannels and the one or more constriction regions have a lipid bilayer disposed thereon. In some embodiments, multiple microfluidic devices operating in parallel are used to process large numbers of cells. The device and method have particularly applicability to delivering gene-editing molecules intracellularly to cells.

High-throughput system and method for the temporary permeabilization of cells using lipid bilayers

A method of transporting biomolecular cargo intracellularly into cells includes the operations of providing magnetic nanostructures (e.g., nanospears, nanostars, nanorods, and other nanometer-sized structures) carrying the biomolecular cargo thereon and applying an external magnetic field to move the magnetic nanostructures into physical contact with at least some of the cells (or the cells into the magnetic nanostructures). The magnetic nanostructures move into physical contact with a single cell, a subset of cells, or all cells. The external magnetic field may be applied by a moving permanent magnet although an electromagnetic may also be used. The biomolecular cargo may include a molecule, a plurality of molecules, or higher order biological constructs. For example, the biological construct may include DNA plasmids, small interfering RNA, proteins, or targeted nuclease gene-editing cargo such as zinc-finger nucleases, transcription activator-like effector nucleases, Cas9 protein, Cas9 mRNA, and associated guide RNA sequences.

Guided magnetic nanostructures for targeted and high-throughput intracellular delivery

A method of detecting and/or discriminating mismatched or complementary nucleic acids using a field-effect transistor (FET). The FET comprises source and drain electrodes formed on substrate and separated by a channel that includes a thin semiconducting film. One or more nucleic acid molecules are immobilized to thin semiconducting film. The FET includes a gate electrode in contact with solution containing the sample (or located on the surface of the device). Samples possibly containing target nucleic acid are exposed to the FET of the biosensor device and current response is measured after samples containing target nucleic acid have been exposed to the FET of the biosensor device. Measured current response in the FET is used to detect and/or discriminate whether target nucleic acid is present as well as complementary or mismatched. Measured current response may also be used to differentiate among different mismatches.

Devices and methods for detecting/discriminating complementary and mismatched nucleic acids using ultrathin film field-effect transistors

Systems and methods are disclosed that utilize metal nanostructures that are synthesized in situ along the internal surfaces of a microfluidic device. The nanostructures are formed by initial deposition of metallic seeds followed by flowing growth and reducing agent solutions into the capillaries/microfluidic channels to grow the nanostars. The nanostructures may optionally be functionalized with a capture ligand. The capture ligand may be used to selectively bind to certain cells (e.g., circulating tumor cells). The cells may be removed by a beam of light (e.g., laser beam) that induces localized heating at the surface location(s) containing the nanostructures. The plasmonic nature of the nanostructures can be used to heat the nanostructure(s) locally for the selective removal of one or certain cells. The nanostructures may be used to acquire Raman spectra of molecules or other small objects that are bound thereto for identification and quantification.

Systems and methods of using anisotropic nanostructures in microfluidic devices for binding and optional release of molecules and cells

A microfluidic device for processing cells for the intracellular delivery of molecules or other cargo includes a plurality of microchannels disposed in a substrate or chip and fluidically coupled to an inlet configured to receive a solution containing the cells and the molecules or other cargo to be delivered intracellularly to the cells. Each of the plurality of microchannels has one or more constriction regions therein, wherein the constriction regions comprise an omniphobic, superhydrophilic, or superhydrophobic surface. In some embodiments, multiple microfluidic devices operating in parallel are used to process large numbers of cells. The device and method has particularly applicability to delivering gene-editing molecules intracellularly to cells.

High-throughput system and method for the temporary permeabilization of cells

A microfluidic-based device and system is disclosed for the high-throughput intracellular delivery of biomolecular cargo to cells (eukaryotic or prokaryotic) or enveloped viruses. Cargo integration occurs due to transient membrane permeabilization by exposure to bulk acoustic waves (BAWs) transduced from surface acoustic waves (SAWs) generated by a rapidly oscillating piezoelectric substrate. In this approach, temporary pores are established across the cellular membrane as cells are partially deformed and squeezed or subject to shearing forces as they travel through the vibrational modes created within the microfludic channel(s) of the device.

Device and method for intracellular delivery of biomolecular cargo via acoustic wave exposure

A robust and general fabrication/manufacturing method is described herein for the fabrication of periodic three-dimensional (3D) hierarchical nanostructures in a highly scalable and tunable manner. This nanofabrication technique exploits the selected and repeated etching of spherical particles that serve as resist material and that can be shaped in parallel for each processing step. The method enables the fabrication of periodic, vertically aligned nanotubes at the wafer scale with nanometer-scale control in three dimensions including outer/inner diameters, heights/hole-depths, and pitches. The method was utilized to construct 3D periodic hierarchical hybrid silicon and hybrid nanostructures such as multi-level solid/hollow nanotowers where the height and diameter of each level of each structure can be configured precisely as well as 3D concentric plasmonic supported metal nanodisk/nanorings with tunable optical properties on a variety of substrates.

Methods for multiple-patterning nanosphere lithography for fabrication of periodic three-dimensional hierarchical nanostructures

The present invention relates to biomimetic membrane compositions and methods for making and using them.

Biomimetic membranes, methods of manufacture and uses thereof

Systems and methods of fabricating and functionality patterned nanowire probes that are stable under fluid reservoir conditions and have imageable contrast are provided. Optical imaging and deconstruction methods and systems are also provided that are capable of determining the distribution of nanowires of a particular pattern to determine the mixing between or leakage from fluid reservoirs.

Determining fluid reservoir connectivity using nanowire probes

Methods and assemblies for the construction of liquid-phase alloy nanoparticles are presented. Particle formation is directed by molecular self-assembly and assisted by sonication. In some embodiments, eutectic gallium-indium (EGaIn) nanoparticles are formed. In these embodiments, the bulk liquid alloy is ultrasonically dispersed, fast thiolate self-assembly at the EGaIn interface protects the material against oxidation. The assembly shell has been designed to include intermolecular hydrogen bonds, which induce surface strain, assisting in cleavage of the alloy particles to the nanoscale. X-ray diffraction and TEM analyses reveal that the nanoscale particles are in an amorphous or liquid phase, with no observed faceting.

Metal alloy nanoparticle synthesis via self-assembled monolayer formation and ultrasound

The present invention is a method and apparatus relating to manufacturing nanostructure patterns and components using molecular science. The method includes overlaying a multilayer organic molecule resist on at least a portion of a parent structure selectively deposited on a substrate, depositing a layer over the parent structure and in contact with at least a portion of the multilayer organic resist, and removing the multilayer organic molecule resist to leave a residual structure.

Molecular ruler for scaling down nanostructures

The present invention discloses devices and methods relating to patterning substrates using chemical functionalization. The methods include covering the surface of a substrate with a first plurality of molecules, selecting at least one internal bond from the plurality of molecules, and reacting the at least one internal bond to form at least one second functional group. Either or both of the functional groups can then be further reacted.

Chemical functionalization nanolithography

A tunable high frequency AC scanning tunneling microscope (ACSTM) has been utilized to image and to record spectra for semiconductor characterization. A difference frequency mixing technique sensitive to dopant type and concentration is applied both to uniformly doped and to patterned semiconductor substrates. Uniformly doped silicon substrates were used to characterize the difference frequency spectral signature for both p- and n-type Si. Comparison of the measured difference frequency to such signature can be used for distinguishing between the two types of dopants in samples with unknown dopant type. Patterned substrates were then fabricated, and a spectroscopic imaging mode was used to map out dopant density at ultrahigh resolution, and to distinguish between areas of different concentration and different dopant type. By measuring samples of known dopant dosages to form a reference database, the unknown dosage of the same dopant in a portion of a sample may be found by comparing the difference signal measured from the sample to the reference database.

Difference frequency imaging and spectroscopy to measure dopants using an alternating current scanning tunneling microscope

A molecular computer is formed by establishing arrays of spaced-apart input and output pins on opposing sides of a containment, injecting moleware in solution into the containment and then allowing the moleware to bridge the input and output pins. Moleware includes molecular alligator clip-bearing 2-, 3-, and molecular 4-, or multi-terminal wires, carbon nanotube wires, molecular resonant tunneling diodes, molecular switches, molecular controllers that can be modulated via external electrical or magnetic fields, massive interconnect stations based on single nanometer-sized particles, and dynamic and static random access memory (DRAM and SRAM) components composed of molecular controller/nanoparticle or fullerene hybrids. The current-voltage characteristics that result from the bridging between input and output arrays can be ascertained using another computer to identify the bundles of inputs and corresponding outputs that provide a truth table for the specific functions of the computer.

Molecular computer

A transducer for detecting biomolecules comprises a bottom and a top unit. The bottom unit has a set of parallel electrically conductive transmission lines thereon and probes attached to selected locations along each transmission line for binding with the target molecule. The top unit has thereon a second set of parallel transmission lines transverse to those on the bottom unit. By applying AC signals sequentially to the two sets of transmission lines, each of the probe locations at the intersection of the two transmission lines can be addressed and the response of the probe in an unfilled detector location or a complex formed by the probe and the target in a filled detector location can be measured. If the target has been labelled, than the label at each of the locations may also be detected. The device can also be used for measuring binding constants between the probe and the target and concentrations of target solutions.

Apparatus and method of detection employing an AC frequency sensor array

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Los Angeles, CA, United States of America

Paul S Weiss