Bio-rad Laboratories, Inc.

10x Genomics, Inc.

Lawrence Livermore National Security, LLC

University of California

Donald Masquelier

Kevin D Ness

Benjamin J Hindson

Billy Wayne Colston, Jr

Serge Saxonov

Christopher Hindson

Rajiv Bharadwaj

Anthony Makarewicz

Fred Paul Milanovich

Debkishore Mitra

Michael Schnall-Levin

Mirna Jarosz

Paul Hardenbol

Christopher F Kelly

Anthony Joseph Makarewicz, Jr

Phillip Belgrader

Amy L Hiddessen

Christopher G Bailey

Erin Rae Chia

Allen T Christian

Shanavaz L Nasarabadi

Perry M Bell

Ladona Willis

Douglas N Modlin

Samuel Burd

Vincent Riot

Billy W Colston

Mark Alan Arbore

Andrew D Price

David L Klein

Tobias Daniel Wheeler

Anthony J Ruggiero

Klaus Willeke

Yuan Li

Jerry E Hurst

Jorge Bernate

Jeffery B Cooke

Jeffery S Kallman

Landon Merrill

A microfluidic system for manipulating particles in a fluid is provided. The system includes a microfluidic chip having at least one channel and at least one filter feature, wherein the filter feature includes a field of physical obstacles configured and arranged to remove physical contaminants associated with the particles. The filter features are particularly suited to remove contaminants associated with gel beads manipulated by microfluidic handling.

Microfluidic on-chip filters

Method of analysis. In the method, a microfluidic device defining a flow path extending from an inlet to an outlet may be selected. A sample-containing fluid may be introduced into the flow path via the inlet. Volumes of the sample-containing fluid may be isolated from one another on the flow path. A two-dimensional monolayer of the volumes may be imaged. The two-dimensional monolayer may be formed along the flow path between the inlet and the outlet.

Method of analysis

Methods of partition-based analysis. In an exemplary method, a device having a port fluidically connected to a chamber may be selected. A sample-containing fluid may be placed into the port. The sample-containing fluid may be moved from the port to the chamber. Partitions of the sample-containing fluid may be formed. A monolayer of the partitions in the chamber may be created. At least a portion of the monolayer may be imaged.

Partition-based method of analysis

A system, including method and apparatus, for generating droplets suitable for droplet-based assays. The disclosed systems may include either one-piece or multi-piece droplet generation components configured to form sample-containing droplets by merging aqueous, sample-containing fluid with a background emulsion fluid such as oil, to form an emulsion of sample-containing droplets suspended in the background fluid. In some cases, the disclosed systems may include channels or other suitable mechanisms configured to transport the sample-containing droplets to an outlet region, so that subsequent assay steps may be performed.

Method of emulsion formation and modification

The disclosure provides devices, systems and methods for the generation of encapsulated reagents and the partitioning of encapsulated reagents for use in subsequent analyses and/or processing, such as in the field of biological analyses and characterization.

Fluidic devices, systems, and methods for encapsulating and partitioning reagents, and applications of same

System for performing a flow-based assay. The system may comprise a droplet generator to produce an emulsion including droplets in a carrier fluid. The system also may comprise a thermocycler including two or more temperature-controlled zones and also including a channel connected to the droplet generator for receiving the emulsion. The channel may form a single-pass continuous fluid route traversing the temperature-controlled zones multiple times, such that droplets passing through the channel are thermally cycled. The system further may comprise a detection station downstream from the thermocycler and configured to detect a signal from the droplets after such droplets have been thermally cycled by passing through the channel.

Droplet-based assay system

Method of analysis. In the method, a first emulsion and a second emulsion substantially separated from one another by a spacer fluid may be formed. The first emulsion, the spacer fluid, and the second emulsion may be flowed in a channel from a fluid inlet to a fluid outlet of a heating and cooling station having two or more temperature-controlled zones, such that each emulsion is thermally cycled to promote amplification of a nucleic acid target in droplets of the emulsion. Amplification data may be collected from individual droplets of each emulsion downstream of the heating and cooling station. A level of the nucleic acid target present in each emulsion may be determined based on the amplification data collected from the individual droplets of the emulsion.

Method of performing droplet-based assays

The disclosure provides methods for separating and/or purifying one or more molecules released from one or more fluid compartments or partitions, such as one or more droplets. Molecules can be released from a fluid compartment(s) and bound to supports that can be isolated via any suitable method, including example methods described herein. The disclosure also provides devices that can aid in isolating supports bound to molecules.

Partition processing methods and systems

This disclosure provides methods and compositions for sample processing, particularly for sequencing applications. Included within this disclosure are bead compositions, such as diverse libraries of beads attached to large numbers of oligonucleotides containing barcodes. Often, the beads provides herein are degradable. For example, they may contain disulfide bonds that are susceptible to reducing agents. The methods provided herein include methods of making libraries of barcoded beads as well as methods of combining the beads with a sample, such as by using a microfluidic device.

Compositions and methods for sample processing

The present disclosure provides compositions, methods, systems, and devices for polynucleotide processing. Such polynucleotide processing may be useful for a variety of applications, including polynucleotide sequencing.

Partitioning and processing of analytes and other species

An integrated system for processing and preparing samples for analysis may include a microfluidic device including a plurality of parallel channel networks for partitioning the samples including various fluids, and connected to a plurality of inlet and outlet reservoirs, at least a portion of the fluids comprising reagents, a holder including a closeable lid hingedly coupled thereto, in which in a closed configuration, the lid secures the microfluidic device in the holder, and in an open configuration, the lid is a stand orienting the microfluidic device at a desired angle to facilitate recovery of partitions or droplets from the partitioned samples generated within the microfluidic device, and an instrument configured to receive the holder and apply a pressure differential between the plurality of inlet and outlet reservoirs to drive fluid movement within the channel networks.

Instrument systems for integrated sample processing

Method of detecting a target nucleic acid. In an exemplary method, at least two thermal zones of different temperature may be created using a heating assembly. A first emulsion and a second emulsion may be formed. The first and second emulsions may be thermally cycled by passing them through tubing in a spaced relation to one another, with the tubing being wound around a central axis of the heating assembly and extending through each thermal zone multiple times. Thermally cycling may promote amplification of the target nucleic acid in droplets of each emulsion. Droplets of each emulsion may be passed through a detection channel located downstream of the tubing. Fluorescence may be detected from the droplets being passed through the detection channel.

Detection method for a target nucleic acid

Method of performing a droplet-based assay. The method may include obtaining droplets encapsulated by an immiscible liquid and packed closely together in a monolayer, performing a reaction in the droplets while packed closely together in the monolayer; and collecting data related to an analyte from a plurality of the droplets while the droplets remain closely packed together in the monolayer.

Device for generating droplets

Growing community of inventors

Tracy, CA, United States of America

Donald Masquelier