University of California

Lawrence Livermore National Security, LLC

Pioneer Hi-Bred International, Inc.

Other

Abraham Phillip Lee

Peter A Krulevitch

William J Benett

M Allen Northrup

Xuan Li

Dennis L Matthews

Joseph P Fitch

Daniel L Schumann

Andrew C Hatch

Duncan J Maitland

Luiz B Da Silva

Yue Yun

Jeffrey S Fisher

Neil Reginald Beer

Peter M Celliers

Hemantha Kumar Wickramasinghe

Dino R Ciarlo

Steven C George

Robert Lin

Charles F McConaghy

Xiaolin Wang

Christopher C W Hughes

Asuncion V Lemoff

Mohammad Aghaamoo

Neha Garg

Do-Hyun Lee

Yinglei Tao

Milton A Northrup

Joe H Satcher, Jr

Stephen M Lane

James Allen Folta

Derek E Decker

Elizabeth K Wheeler

Armando R Tovar

Richard A London

David S Clague

Christopher B Darrow

Michelle A Digman

Amy W Wang

Karla G Hagans

Michael E Glinsky

Robert E Clough

Yu-Hsi Chen

Duc Phan

Dohyun Lee

Gopakumar Kamalakshakurup

Melinda G Simon

Javier Lopez-Prieto

Maulik V Patel

Charles A Jungreis

Peter Krulevich

Jimmy C Trevino

Philip L Felgner

Mark Bachman

Gary E Sommargren

Robert J Deri

Robin R Miles

Michael D Pocha

John Collins

Luiz B DaSilva

Roger Shih

Yu-Hsiang Hsu

Ning Ma

Edwin S Monuki

Naiqing Zhang

Derek Vellejo

Crystal Rapier

Arlene Doria

Nicholas E Martin

Yuka Okabe

Uland Liao

Javier L Prieto

Monica Moya

Jente Lu

Lisa A Flanagan

Mindy Simon

Nick Martin

Lisen Wang

Lisa Flanagan Monuki

Svyatoslav A Marchenko

Yung-Cheih Tan

Albert Tsung-Hsi Hsieh

Yung-Chieh Tan

Tao Yue

Da Zhao

Christopher C Hughes

Gisela Lin

Xuhao Luo

Marzieh Ataei

Francesco Palomba

Wei-Feng Fang

Jonathon N Simon

Paul E Ahre

Peter C Dupuy

The present invention features the use of cavity acoustic transducers (CATs) to apply mechanical stimuli on cells. CATs utilize the generated acoustic microstreaming vortices to trap cells and apply tunable shear on them. The present invention may use such a portable, automated, and high throughput device for cell transfection.

Cavity acoustic transducer (CAT) for shear-induced cell transfection

A microfluidic device having an array for cell trapping is used to analyze cell-cell interaction at single-cell level. The microfluidic trapping array efficiently pairs single cells in isolated compartments in an easy-to-operate manner. A first cell is squeezed through an opening of a first cavity by a strong forward flow. Subsequently, a second cell is pushed into a second cavity by a low reverse flow. The trapped cell pairs are sealed by an oil phase or hydrogel into isolated compartments, thereby eliminating interference from other cell pairs or the surrounding media.

Microfluidic single-cell pairing array for studying cell-cell interactions in isolated compartments

A pressure regulator module for a chip-based microfluidic platform is provided. The module includes a microfluidic channel for passing flowable material from the inlet region through the outlet region and into a downstream compartment; one or more microvalves fluidly connected to the microfluidic channel and upstream of the outlet region; and one or more reservoirs fluidly connected to the microvalves, for receiving flowable material diverted by the microvalves, where a flow of flowable material passing from the inlet region toward the downstream compartment is at least partially diverted by the microvalves into the reservoirs as a result of a pressure increase in the microfluidic channel. In some versions, the microvalves are capillary burst valves. A microfluidic chip containing the module and a method of using the module are provided.

Microfluidic pressure regulator for robust hydrogel loading without bursting

Methods and devices for single cell analysis using fluorescence lifetime imaging microscopy (FLIM) are disclosed. The methods utilize microfluidic devices which use traps to immobilize cells for FLIM analysis. The analysed cells may be sorted before or after imaging and may be plant, animal, or bacterial cells. Analysis of the FLIM data may use a phasor plot and may be used to identify a metabolic pattern of the single cells.

Microfluidic label-free isolation and identification of cells using fluorescence lifetime imaging (FLIM)

Systems and methods for transfection using a microfluidic device are disclosed. Microdroplets encapsulate cells, transfection molecules, and cationic lipid transfection reagent. Droplet chaotic advection in a rendering channel of the system results in a uniform lipid-DNA complex (lipoplex) formation, which can improve gene delivery efficacy. The shear stress exerted on cell membranes during the chaotic mixing increases membrane permeability, which when combined with the co-confinement of cell and lipoplex, improves transfection efficiency of the cell. The systems and methods can be used for a variety of applications such as gene therapy, in vitro fertilization, regenerative medicine, cancer treatment, and vaccines.

Microfluidic systems and methods for lipoplex-mediated cell transfection

A method of encapsulating a solid sample in a droplet, the method including flowing a continuous phase through a first fluid channel at a first flow rate; flowing a dispersed phase through a second fluid channel at a second flow rate, the dispersed phase including a plurality of particles, cells or beads; trapping the plurality of particles, cells or beads in a mixing region that receives the dispersed phase and the continuous phase; and reducing the first flow rate to encapsulate the trapped particles, cells or beads in droplets of the dispersed phase generated when the dispersed phase and the continuous phase exit the mixing region through an orifice.

Enhanced cell/bead encapsulation methods and apparatuses

A passive, hydrodynamic technique implemented using a microfluidic device to perform co-encapsulation of samples in droplets and sorting of said droplets is described herein. The hydrodynamic technique utilizes laminar flows and high shear liquid-liquid interfaces at a microfluidic junction to encapsulate samples in the droplets. A sorting mechanism is implemented to separate sample droplets from empty droplets. This technique can achieve a one-one-one encapsulation efficiency of about 80% and can significantly improve the droplet sequencing and related applications in single cell genomics and proteomics.

High-efficiency particle encapsulation in droplets with particle spacing and downstream droplet sorting

A multi-layered microfluidic system featuring tissue chambers for cells in a first layer and a plurality of medium channels for culture medium in a second layer. The tissue chambers fluidly connect to the medium channels such that media flows from the medium channels to the tissue chambers, forming large-scale perfused capillary networks. The capillary networks can undergo angiogenesis and vertical anastomosis. The multi-layered configuration of the system of the present invention allows for flexibility in design.

Multi-layered microfluidic systems for in vitro large-scale perfused capillary networks

A method of trapping constituents of interest in a fluid sample flowing through a microfluidic channel by vibrating an interface of the fluid sample and a gas occupying a lateral channel adjacent the microfluidic channel is described. A marker is flowed into the microfluidic channel such that the marker bonds with constituents of interest. The constituents of interest bonded to the marker can help identification of the constituents of interest.

Microfluidic in situ labelling on stable interfaces

Microfluidic systems and methods to generate and analyze microcapsules comprising biological sample, such as for example, single cells, cellular contents, microspore, protoplast, are disclosed. The microcapsules comprising the biological sample can be preserved by a polymerization process that forms a hydrogel around the biological sample. The hydrogel microcapsules can be trapped in a trapping array or collected in an output reservoir and subject to one or more assays. The trapping array or the output reservoir can be disposed over a porous layer that can filter the continuous phase (e.g., oil) in which the microcapsules are dispersed in the microfluidic device. The pores of the porous layer are configured to be smaller than the size of the microcapsules to prevent the flow of the microcapsules through the porous layer.

Systems and methods to encapsulate and preserve organic matter for analysis

The present invention features the use of lateral cavity acoustic transducers (LCATs) to apply mechanical stimuli on cells. LCATs utilize the generated acoustic microstreaming vortices to trap cells and apply tunable shear-induced cell deformation on them. The present invention may use such a portable, automated, and high throughput device for shear-induced cell transfection.

Lateral cavity acoustic transducer (LCAT) for shear-induced cell transfection

Disclosed is a water-in-oil-in-water (W/O/W) double emulsion including a first water phase, an oil phase and a second water phase, wherein the W/O/W double emulsion is disposed in an isotonic solution, and related methods of making the W/O/W double emulsion. Also disclosed is a method of making an artificial antigen presenting cell including: providing a W/O/W double emulsion that is stored in an isotonic solution, wherein the W/O/W double emulsion includes a peptide associated with a Major Histocompatibility (pMHC) complex or a glycolipid antigen associated with a CD1d molecule, and a costimulatory molecule; and transferring the W/O/W double emulsion to an electrolyte solution, wherein the double emulsion undergoes a morphological transformation to become the artificial antigen presenting cell. Also disclosed is a method of drug delivery including administering to a subject a unilamellar vesicle containing the drug. Other methods relate to storing a protein including making a water-in-oil-in-water (W/O/W) double emulsion, wherein the W/O/W double emulsion includes the protein, and wherein the W/O/W double emulsion is configured to be stably stored.

On demand vesicle formation from vesicle precursors suitable for long-term storage

An interfacial technique utilizes hydrodynamic micro-vortices to perform (i) high efficiency single cell encapsulation and (ii) size-selective capturing of cells based on their sizes in a single microfluidic device. A notable feature of this technique is that it can perform high efficiency single cell encapsulation at low cell concentrations, and this technique is all passive, controlled only by the flow rates of the two phases and does not require complex structures or on-chip active devices. Single bead/cell encapsulation was demonstrated at 50% efficiency, which is at least 10 times greater than the random encapsulations at the introduced cell concentrations. Also demonstrated is the selective trapping of cells based on their sizes. This present technique expands the capabilities of droplet microfluidics for applications ranging from single cell genomics, proteomic assays to sample preparation.

High-efficiency encapsulation in droplets based on hydrodynamic vortices control

Novel systems and methods are provided that rapidly separate particles from a liquid. In an embodiment, a small volume of liquid (such as a blood sample, or any other solution with a concentration of particles) is input into a flow device implemented as a unilateral channel. When activated by an acoustic energy source (such as an ultrasound pulse), gas-liquid interfaces naturally occurring between the liquid in the flow device and a plurality of gas-filled cavities that line the channel will oscillate and create stable cavitation streaming within a localized region of the surrounding liquid. These oscillations create micro-vortices that gently remove and trap particles and debris from the liquid and adjacent surfaces. Fluid and particle manipulation can thus be accomplished on a passive, disposable chip that is placed on top of an external acoustic transducer with a coupling medium.

Microfluidic systems for particle trapping and separation using cavity acoustic transducers

Growing community of inventors

Irvine, CA, United States of America

Abraham Phillip Lee