Described herein are microparticles each comprising a plurality of bound biological molecules. Further described herein is a plurality of microdroplets each comprising one or more primer vehicles. Methods of making and using these microdroplets are also reported. An exemplary microparticle is of Formula (I).
The invention includes methods for determining the presence of a latent viral population by analyzing an RNA population from the virus with digital techniques, such as digital PCR or by sequencing cDNA produced from the RNA. The invention additional includes methods for determining the presence of latent viral populations by detecting and/or quantifying enzymes that are uniquely associated with the virus, e.g., reverse transcriptases.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C12Q 1/70 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving virus or bacteriophage
C12P 19/34 - Polynucleotides, e.g. nucleic acids, oligoribonucleotides
3.
Manipulation of fluids and reactions in microfluidic systems
Microfluidic structures and methods for manipulating fluids and reactions are provided. Such structures and methods may involve positioning fluid samples, e.g., in the form of droplets, in a carrier fluid (e.g., an oil, which may be immiscible with the fluid sample) in predetermined regions in a microfluidic network. In some embodiments, positioning of the droplets can take place in the order in which they are introduced into the microfluidic network (e.g., sequentially) without significant physical contact between the droplets. Because of the little or no contact between the droplets, there may be little or no coalescence between the droplets. Accordingly, in some such embodiments, surfactants are not required in either the fluid sample or the carrier fluid to prevent coalescence of the droplets. Structures and methods described herein also enable droplets to be removed sequentially from the predetermined regions.
C40B 50/08 - Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creationParticular methods of cleavage from the liquid support
The invention generally relates to droplet based digital PCR and methods for analyzing a target nucleic acid using the same. In certain embodiments, methods of the invention involve forming sample droplets containing, on average, a single target nucleic acid, amplifying the target in the droplets, excluding droplets containing amplicon from the target and amplicon from a variant of the target, and analyzing target amplicons.
Assemblies for displacing droplets from a vessel that facilitate the collection and transfer of the droplets while minimizing sample loss. The assembly includes at least one droplet formation module, in which the module is configured to form droplets surrounded by an immiscible fluid. The assembly includes at least one chamber including an outlet, in which the chamber is configured to receive droplets and an immiscible fluid, and in which the outlet is configured to receive substantially only droplets. The assembly further includes a channel, configured such that the droplet formation module and the chamber are in fluid communication with each other via the channel. The assembly includes a plurality of hollow members, in which the hollow members are channels and in which the members are configured to interact with a vessel. The assembly includes a main channel, wherein the second member is in fluid communication with the main channel.
B01F 5/06 - Mixers in which the components are pressed together through slits, orifices, or screens
B01J 2/02 - Processes or devices for granulating materials, in generalRendering particulate materials free flowing in general, e.g. making them hydrophobic by dividing the liquid material into drops, e.g. by spraying, and solidifying the drops
B81B 1/00 - Devices without movable or flexible elements, e.g. microcapillary devices
G01N 35/08 - Automatic analysis not limited to methods or materials provided for in any single one of groups Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions
8.
Manipulation of fluids and reactions in microfluidic systems
Microfluidic structures and methods for manipulating fluids and reactions are provided. Such structures and methods may involve positioning fluid samples, e.g., in the form of droplets, in a carrier fluid (e.g., an oil, which may be immiscible with the fluid sample) in predetermined regions in a microfluidic network. In some embodiments, positioning of the droplets can take place in the order in which they are introduced into the microfluidic network (e.g., sequentially) without significant physical contact between the droplets. Because of the little or no contact between the droplets, there may be little or no coalescence between the droplets. Accordingly, in some such embodiments, surfactants are not required in either the fluid sample or the carrier fluid to prevent coalescence of the droplets. Structures and methods described herein also enable droplets to be removed sequentially from the predetermined regions.
The invention generally relates to droplet based digital PCR and methods for analyzing a target nucleic acid using the same. In certain embodiments, a method for determining the nucleic acid make-up of a sample is provided. In one aspect, the invention provides a droplet that contains a single nucleic acid template and a plurality of primer pairs specific for multiple target sites on the template. The single nucleic acid template can be DNA or RNA. The template is amplified in the droplet for detection; and may preferably be amplified using a plurality of primer pairs as described herein. The ability to amplify and detect single nucleic acids in droplets enables digital PCR, detection, counting, and differentiation among nucleic acids, especially those present in heterogeneous samples. Thus, the invention applies to digital amplification techniques and, in specific embodiments enables multiplex PCR in droplets.
09 - Scientific and electric apparatus and instruments
Goods & Services
Scientific apparatus, instruments and equipment for scientific research, non-medical diagnostic analysis, chemical analysis, clinical analysis and industrial use, namely, DNA and RNA testing apparatus and genetic analyzers.
09 - Scientific and electric apparatus and instruments
Goods & Services
Scientific apparatus, instruments and equipment for scientific research, non-medical diagnostic analysis, chemical analysis, clinical analysis and industrial use, namely, DNA and RNA testing apparatus, namely, genetic analyzers, and polymerase chain reaction (PCR) cyclers and samplers.
The invention generally relates to method for screening for a condition in a subject. In certain embodiments, methods of the invention involve obtaining a pool of nucleic acids from a sample, incubating the nucleic acids with first and second sets of binders, in which the first set binds uniquely to different regions of a target nucleic acid in the pool, the second set binds uniquely to different regions of a reference nucleic acid in the pool, and the first and second sets include different detectable labels, removing unbound binders, detecting the labels, and screening for a condition based upon results of the detecting step.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
C07H 21/00 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
The invention generally relates to methods for quantifying an amount of enzyme molecules. Systems and methods of the invention are provided for measuring an amount of target by forming a plurality of fluid partitions, a subset of which include the target, performing an enzyme-catalyzed reaction in the subset, and detecting the number of partitions in the subset. The amount of target can be determined based on the detected number.
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions
The invention provides barcode libraries and methods of making and using them including obtaining a plurality of nucleic acid constructs in which each construct comprises a unique N-mer and a functional N-mer and segregating the constructs into a fluid compartments such that each compartment contains one or more copies of a unique construct. The invention further provides methods for digital PCR and for use of barcode libraries in digital PCR.
C07H 21/00 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
C40B 40/06 - Libraries containing nucleotides or polynucleotides, or derivatives thereof
Methods involve forming a droplet, and contacting the droplet with a fluid stream, wherein a portion of the fluid stream integrates with the droplet to form a mixed droplet. Methods for merging two liquid phases in which only one phase is in the form of a droplet at least at the point of merging A second phase is injected into the drops directly from a continuous stream. Methods of the invention provide a simple and reliable approach to sample fluid mixing because only one of the two phases is dispersed as a droplet prior to its merge with the other phase.
The present invention provides thermocycling devices useful for amplification of nucleic acids in droplets. The thermocycling device utilizes the flow of one or more fluids through a main compartment at temperatures sufficient to conduct a polymerase chain reaction. Methods of amplifying nucleic acids in droplets are also provided.
Methods of performing sandwich assays in droplets are disclosed. Further disclosed are methods for detecting a target analyte that involve forming a compartmentalized portion of fluid including a portion of a sample suspected of containing a target analyte and a sample identifier, a first binding agent having a target identifier, and a second binding agent specific to the target analyte under conditions that produce a complex of the first and second binding agents with the target analyte; separating the complexes, and detecting the complexes, thereby detecting the target analyte.
The invention generally relates to droplet based digital PCR and methods for analyzing a target nucleic acid using the same. In certain embodiments, methods of the invention involve forming sample droplets containing, on average, a single target nucleic acid, amplifying the target in the droplets, excluding droplets containing amplicon from the target and amplicon from a variant of the target, and analyzing target amplicons.
The present invention generally relates to systems and methods to create stable emulsions with low rates of exchange of molecules between microdroplets.
G01N 35/08 - Automatic analysis not limited to methods or materials provided for in any single one of groups Handling materials therefor using a stream of discrete samples flowing along a tube system, e.g. flow injection analysis
G01N 33/48 - Biological material, e.g. blood, urineHaemocytometers
21.
LABELLED SILICA-BASED NANOMATERIAL WITH ENHANCED PROPERTIES AND USES THEREOF
The invention provides methods for assessing one or more predetermined characteristics or properties of a microfluidic droplet within a microfluidic channel, and regulating one or more fluid flow rates within that channel to selectively alter the predetermined microdroplet characteristic or property using a feedback control. The assessment of the characteristics can be made using an image sensor.
01 - Chemical and biological materials for industrial, scientific and agricultural use
09 - Scientific and electric apparatus and instruments
44 - Medical, veterinary, hygienic and cosmetic services; agriculture, horticulture and forestry services
Goods & Services
Reagents being part of a droplet based high throughput chemical compound screening system, namely a compound screening system and in vitro testing for research purposes. Computer hardware, software, microfluidic chip, being part of a droplet based high throughput chemical compound screening system. Droplet based high throughput drug screening and in vitro testing services.
The present invention generally relates to droplet libraries and to systems and methods for the formation of libraries of droplets. The present invention also relates to methods utilizing these droplet libraries in various biological, chemical, or diagnostic assays.
C40B 40/04 - Libraries containing only organic compounds
C40B 40/06 - Libraries containing nucleotides or polynucleotides, or derivatives thereof
C40B 50/08 - Liquid phase synthesis, i.e. wherein all library building blocks are in liquid phase or in solution during library creationParticular methods of cleavage from the liquid support
09 - Scientific and electric apparatus and instruments
Goods & Services
Droplet based microfluidics system comprised of computer hardware, software, reagents and a microfluidic chip for conducting biomedical research and molecular diagnostics that includes genomic and proteomic analysis, pharmaceutical compound screening, cellular analysis and sorting and biomarker detection and parts and attachments therefore.
The present invention provides novel microfluidic substrates and methods that are useful for performing biological, chemical and diagnostic assays. The substrates can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged such that a continuous channel is provided for flow of immiscible fluids.
42 - Scientific, technological and industrial services, research and design
Goods & Services
Commercialization of platforms for analyzing and manipulating fluids for life science research, cancer diagnosis and the detection of residual disease. Research and development of platforms for analyzing and manipulating fluids for life science research, cancer diagnosis and the detection of residual disease.
LE CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE (France)
RAINDANCE TECHNOLOGIES, INC. (USA)
Inventor
Holtze, Christian
Guerra, Rodrigo, E.
Agresti, Jeremy
Weitz, David, A.
Ahn, Kuenho
Hutchison, John, Brian
Griffiths, Andrew
El Harrak, Abdeslam
Miller, Oliver, Jon
Baret, Jean-Christophe
Taly, Valérie
Ryckelynck, Michaël
Merten, Christoph
Abstract
Surfactants (e.g., fluorosurfactants) for stabilizing aqueous or hydrocarbon droplets in a fluorophilic continuous phase are presented. In some embodiments, fluorosurfactants include a fluorophilic tail soluble in a fluorophilic (e.g., fluorocarbon) continuous phase, and a headgroup soluble in either an aqueous phase or a lipophilic (e.g., hydrocarbon) phase. The combination of a fluorophilic tail and a headgroup may be chosen so as to create a surfactant with a suitable geometry for forming stabilized reverse emulsion droplets having a disperse aqueous or lipophilic phase in a continuous, fluorophilic phase. In some embodiments, the headgroup is preferably non-ionic and can prevent or limit the adsorption of molecules at the interface between the surfactant and the discontinuous phase. This configuration can allow the droplet to serve, for example, as a reaction site for certain chemical and/or biological reactions. In another embodiment, aqueous droplets are stabilized in a fluorocarbon phase at least in part by the electrostatic attraction of two oppositely charged or polar components, one of which is at least partially soluble in the dispersed phase, the other at least partially soluble in the continuous phase. One component may provide colloidal stability of the emulsion, and the other may prevent the adsorption of biomolecules at the interface between a component and the discontinuous phase. Advantageously, surfactants and surfactant combinations of the invention may provide sufficient stabilization against coalescence of droplets, without interfering with processes that can be carried out inside the droplets.
The present invention provides novel microfluidic substrates and methods that are useful for performing biological, chemical and diagnostic assays. The substrates can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged such that a continuous channel is provided for flow of immiscible fluids.
The present invention provides novel microfluidic devices and methods that are useful for performing high-throughput screening assays and combinatorial chemistry. Such methods can include labeling a library of compounds by emulsifying aqueous solutions of the compounds and aqueous solutions of unique liquid labels on a microfluidic device, which includes a plurality of electrically addressable, channel bearing fluidic modules integrally arranged on a microfabricated substrate such that a continuous channel is provided for flow of immiscible fluids, whereby each compound is labeled with a unique liquid label, pooling the labeled emulsions, coalescing the labeled emulsions with emulsions containing a specific cell or enzyme, thereby forming a nanoreactor, screening the nanoreactors for a desirable reaction between the contents of the nanoreactor, and decoding the liquid label, thereby identifying a single compound from a library of compounds.
The present invention provides novel microfluidic devices and methods that are useful for performing high-throughput screening assays and combinatorial chemistry. The device can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged on a microfabricated substrate such that a continuous channel is provided for flow of immiscible fluids.
The present invention provides novel microfluidic devices, kits, and methods that are useful for performing high-throughput screening assays and diagnostics. Such diagnostic methods can include emulsifying an aqueous library of compounds with a set of uniquely dyecoded-labeled droplets in an inert fluorocarbon medium, thereby forming an interactor library, emulsifying an aqueous sample from a subject in an inert fluorocarbon medium, wherein said sample contains a compound that will react with at least one interactor molecule from the interactor library, coalescing the emulsions to form a nanoreactor, and screening the nanoreactors for a desirable reaction between the contents of the nanoreactor, herein one or more steps are performed on a microfluidic device, which can include a plurality of electrically addressable, channel bearing fluidic modules integrally arranged on a microfabricated substrate such that a continuous channel is provided for flow of immiscible fluids.
