The present invention relates to droplet-based surface modification and washing. According to one embodiment, a method of splitting a droplet is provided, the method including providing a droplet microactuator including a droplet including one or more beads and immobilizing at least one of the one or more beads. The method further includes conducting one or more droplet operations to divide the droplet to yield a set of droplets including a droplet including the one or more immobilized beads and a droplet substantially lacking the one or more immobilized beads.
G01N 27/26 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variablesInvestigating or analysing materials by the use of electric, electrochemical, or magnetic means by using electrolysis or electrophoresis
B01F 33/302 - Micromixers the materials to be mixed flowing in the form of droplets
B01F 33/3031 - Micromixers using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
A method comprising effecting a change in a shape of a droplet, wherein the droplet is disposed over a substrate in sensing proximity to a sensor and the droplet has a starting surface area exposed to the sensor; and producing an expanded surface area of the droplet in the sensing proximity exposed to the sensor, wherein the expanded surface area exposed to the sensor is greater than the starting surface area exposed to the sensor.
Methods are provided for separating magnetically responsive beads from a droplet in a droplet actuator. Droplet operations electrodes and a magnet are arranged in a droplet actuator to manipulate a bead-containing droplet and position it relative to a magnetic field region that attracts the magnetically responsive beads. The droplet operations electrodes are operated to control the droplet shape and transport it away from the magnetic field region to form a concentration of beads in the droplet. The continued transport of the droplet away from the magnetic field causes the concentration of beads to break away from the droplet to yield a small, concentrated bead-containing droplet immobilized by the magnet.
B01F 31/65 - Mixers with shaking, oscillating, or vibrating mechanisms the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
B01F 33/302 - Micromixers the materials to be mixed flowing in the form of droplets
B01F 33/3031 - Micromixers using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
4.
Methods of improving accuracy and precision of droplet metering using an on-actuator reservoir as the fluid input
Methods of improving accuracy of droplet metering using at least one on-actuator reservoir as the fluid input. In some embodiments, the on-actuator reservoir that is used for metering droplets includes a loading port, a liquid storage zone, a droplet metering zone, and a droplet dispensing zone. The on-actuator reservoirs are designed to prevent liquid flow-back into the loading port and to prevent liquid from flooding into the droplet operations gap in the dispensing zone.
B05B 5/025 - Discharge apparatus, e.g. electrostatic spray guns
G01F 23/22 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water
G01F 23/24 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of resistance of resistors due to contact with conductor fluid
G01F 23/26 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of capacity or inductance of capacitors or inductors arising from the presence of liquid or fluent solid material in the electric or electromagnetic fields
B05B 12/08 - Arrangements for controlling deliveryArrangements for controlling the spray area responsive to condition of liquid or other fluent material discharged, of ambient medium or of target
The present invention relates to droplet-based surface modification and washing. According to one embodiment, a method of splitting a droplet is provided, the method including providing a droplet microactuator including a droplet including one or more beads and immobilizing at least one of the one or more beads. The method further includes conducting one or more droplet operations to divide the droplet to yield a set of droplets including a droplet including the one or more immobilized beads and a droplet substantially lacking the one or more immobilized beads.
G01N 27/26 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variablesInvestigating or analysing materials by the use of electric, electrochemical, or magnetic means by using electrolysis or electrophoresis
B01F 33/302 - Micromixers the materials to be mixed flowing in the form of droplets
B01F 33/3031 - Micromixers using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
B01L 7/00 - Heating or cooling apparatusHeat insulating devices
The present invention is directed to methods of improving accuracy of droplet metering using at least one on-actuator reservoir as the fluid input. In some embodiments, the on-actuator reservoir that is used for metering droplets includes a loading port, a liquid storage zone, a droplet metering zone, and a droplet dispensing zone. The on-actuator reservoirs are designed to prevent liquid flow-back into the loading port and to prevent liquid from flooding into the droplet operations gap in the dispensing zone.
B05B 5/025 - Discharge apparatus, e.g. electrostatic spray guns
G01F 23/22 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water
G01F 23/24 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of resistance of resistors due to contact with conductor fluid
G01F 23/26 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of capacity or inductance of capacitors or inductors arising from the presence of liquid or fluent solid material in the electric or electromagnetic fields
B05B 12/08 - Arrangements for controlling deliveryArrangements for controlling the spray area responsive to condition of liquid or other fluent material discharged, of ambient medium or of target
7.
Manipulation of beads in droplets and methods for manipulating droplets
Provided herein are methods of splitting droplets containing magnetically responsive beads in a droplet actuator. A droplet actuator having a plurality of droplet operations electrodes configured to transport the droplet, and a magnetic field present at the droplet operations electrodes, is provided. The magnetically responsive beads in the droplet are immobilized using the magnetic field and the plurality of droplet operations electrodes are used to split the droplet into first and second droplets while the magnetically responsive beads remain substantially immobilized.
G01N 33/543 - ImmunoassayBiospecific binding assayMaterials therefor with an insoluble carrier for immobilising immunochemicals
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
B01F 31/65 - Mixers with shaking, oscillating, or vibrating mechanisms the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
B01F 33/451 - Magnetic mixersMixers with magnetically driven stirrers wherein the mixture is directly exposed to an electromagnetic field without use of a stirrer, e.g. for material comprising ferromagnetic particles or for molten metal
B01F 33/302 - Micromixers the materials to be mixed flowing in the form of droplets
B01F 33/3031 - Micromixers using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
G01N 1/38 - Diluting, dispersing or mixing samples
B01L 7/00 - Heating or cooling apparatusHeat insulating devices
G01N 35/10 - Devices for transferring samples to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
Methods are provided for separating magnetically responsive beads from a droplet in a droplet actuator. Droplet operations electrodes and a magnet are arranged in a droplet actuator to manipulate a bead-containing droplet and position it relative to a magnetic field region that attracts the magnetically responsive beads. The droplet operations electrodes are operated to control the droplet shape and transport it away from the magnetic field region to form a concentration of beads in the droplet. The continued transport of the droplet away from the magnetic field causes the concentration of beads to break away from the droplet to yield a small, concentrated bead-containing droplet immobilized by the magnet.
B01F 31/65 - Mixers with shaking, oscillating, or vibrating mechanisms the materials to be mixed being directly submitted to a pulsating movement, e.g. by means of an oscillating piston or air column
B01F 33/302 - Micromixers the materials to be mixed flowing in the form of droplets
B01F 33/3031 - Micromixers using electro-hydrodynamic [EHD] or electro-kinetic [EKI] phenomena to mix or move the fluids
B01L 7/00 - Heating or cooling apparatusHeat insulating devices
G01N 35/10 - Devices for transferring samples to, in, or from, the analysis apparatus, e.g. suction devices, injection devices
The present invention relates to droplet-based surface modification and washing. According to one embodiment, a method of splitting a droplet is provided, the method including providing a droplet microactuator including a droplet including one or more beads and immobilizing at least one of the one or more beads. The method further includes conducting one or more droplet operations to divide the droplet to yield a set of droplets including a droplet including the one or more immobilized beads and a droplet substantially lacking the one or more immobilized beads.
G01N 27/26 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variablesInvestigating or analysing materials by the use of electric, electrochemical, or magnetic means by using electrolysis or electrophoresis
B01F 13/00 - Other mixers; Mixing plant, including combinations of dissimilar mixers
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
B01L 7/00 - Heating or cooling apparatusHeat insulating devices
G01N 35/00 - Automatic analysis not limited to methods or materials provided for in any single one of groups Handling materials therefor
A method comprising effecting a change in a shape of a droplet, wherein the droplet is disposed over a substrate in sensing proximity to a sensor and the droplet has a starting surface area exposed to the sensor; and producing an expanded surface area of the droplet in the sensing proximity exposed to the sensor, wherein the expanded surface area exposed to the sensor is greater than the starting surface area exposed to the sensor.
Provided herein are methods of splitting droplets containing magnetically responsive beads in a droplet actuator. A droplet actuator having a plurality of droplet operations electrodes configured to transport the droplet, and a magnetic field present at the droplet operations electrodes, is provided. The magnetically responsive beads in the droplet are immobilized using the magnetic field and the plurality of droplet operations electrodes are used to split the droplet into first and second droplets while the magnetically responsive beads remain substantially immobilized.
The present invention is directed to methods of improving accuracy of droplet metering using at least one on-actuator reservoir as the fluid input. In some embodiments, the on-actuator reservoir that is used for metering droplets includes a loading port, a liquid storage zone, a droplet metering zone, and a droplet dispensing zone. The on-actuator reservoirs are designed to prevent liquid flow-back into the loading port and to prevent liquid from flooding into the droplet operations gap in the dispensing zone.
B05B 5/025 - Discharge apparatus, e.g. electrostatic spray guns
G01F 23/22 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water
G01F 23/24 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of resistance of resistors due to contact with conductor fluid
G01F 23/26 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of capacity or inductance of capacitors or inductors arising from the presence of liquid or fluent solid material in the electric or electromagnetic fields
B05B 12/08 - Arrangements for controlling deliveryArrangements for controlling the spray area responsive to condition of liquid or other fluent material discharged, of ambient medium or of target
Methods are provided for separating magnetically responsive beads from a droplet in a droplet actuator. Droplet operations electrodes and a magnet are arranged in a droplet actuator to manipulate a bead-containing droplet and position it relative to a magnetic field region that attracts the magnetically responsive beads. The droplet operations electrodes are operated to control the droplet shape and transport it away from the magnetic field region to form a concentration of beads in the droplet. The continued transport of the droplet away from the magnetic field causes the concentration of beads to break away from the droplet to yield a small, concentrated bead-containing droplet immobilized by the magnet.
The invention provides a method of circulating magnetically responsive beads within a droplet in a droplet actuator. The invention also provides methods for splitting droplets. The invention, in one embodiment, makes use of a droplet actuator with top and bottom substrates, a plurality of magnetic fields respectively present proximate the top and bottom substrates, wherein at least one of the magnet fields is selectively alterable, and a plurality of droplet operations electrodes positioned along at least one of the top and bottom surfaces. A droplet is positioned between the top and bottom surfaces and at least one of the magnetic fields is selectively altered.
The invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay.
The invention relates to a droplet actuator device and methods for integrated sample preparation and analysis of a biological sample. A droplet actuator device is provided for conducting droplet operations. The droplet actuator device may include a bottom substrate and a top substrate separated from each other to form a gap therebetween; an arrangement of droplet operations electrodes arranged on one or both of the bottom and/or top substrates for conducting droplet operations thereon; a reagent storage layer comprising one or more compartments bound to the top substrate; and one or more openings arranged to provide a fluidic path from the one or more compartments into the gap, upon breach of a breachable seal separating the one or more compartments and openings.
The present invention is directed to methods of improving accuracy of droplet metering using at least one on-actuator reservoir as the fluid input. In some embodiments, the on-actuator reservoir that is used for metering droplets includes a loading port, a liquid storage zone, a droplet metering zone, and a droplet dispensing zone. The on-actuator reservoirs are designed to prevent liquid flow-back into the loading port and to prevent liquid from flooding into the droplet operations gap in the dispensing zone.
B05B 1/08 - Nozzles, spray heads or other outlets, with or without auxiliary devices such as valves, heating means designed to produce a jet, spray, or other discharge of particular shape or nature, e.g. in single drops of pulsating nature, e.g. delivering liquid in successive separate quantities
B05B 5/025 - Discharge apparatus, e.g. electrostatic spray guns
G01F 23/22 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water
G01F 23/24 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of resistance of resistors due to contact with conductor fluid
G01F 23/26 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of capacity or inductance of capacitors or inductors arising from the presence of liquid or fluent solid material in the electric or electromagnetic fields
B05B 12/08 - Arrangements for controlling deliveryArrangements for controlling the spray area responsive to condition of liquid or other fluent material discharged, of ambient medium or of target
19.
INSTRUMENT AND CARTRIDGE FOR PERFORMING ASSAYS IN A CLOSED SAMPLE PREPARATION AND REACTION SYSTEM EMPLOYING ELECTROWETTING FLUID MANIPULATION
In one embodiment, a multiplex fluid processing cartridge includes a sample well, a deformable fluid chamber, a mixing well with a mixer disposed therein, a lysis chamber including a lysis mixer, an electrowetting grid for microdroplet manipulation, and electrosensor arrays configured to detect analytes of interest. An instrument for processing the cartridge is configured to receive the cartridge and to selectively apply thermal energy, magnetic force, and electrical connections to one or more discrete locations on the cartridge and is further configured to compress the deformable chamber(s) in a specified sequence.
The invention provides a method of circulating magnetically responsive beads within a droplet in a droplet actuator. The invention also provides methods for splitting droplets. The invention, in one embodiment, makes use of a droplet actuator with top and bottom substrates, a plurality of magnetic fields respectively present proximate the top and bottom substrates, wherein at least one of the magnet fields is selectively alterable, and a plurality of droplet operations electrodes positioned along at least one of the top and bottom surfaces. A droplet is positioned between the top and bottom surfaces and at least one of the magnetic fields is selectively altered.
A method, circuit and apparatus for detecting capacitance on a droplet actuator, inter alia, for determining the presence, partial presence or absence of a droplet at an electrode on a droplet actuator by: (a) providing a droplet actuator comprising: (i) a substrate comprising electrodes arranged on the substrate for conducting droplet operations on a surface of the substrate; (ii) a capacitance detection circuit for detecting capacitance at the droplet operations surface at one or more of the electrodes; (b) detecting capacitance at the droplet operations surface at one or more of the electrodes; and (c) determining from the capacitance the presence, partial presence or absence of a droplet at the droplet operations surface at the electrode.
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
G01N 27/22 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance
G01V 3/02 - Electric or magnetic prospecting or detectingMeasuring magnetic field characteristics of the earth, e.g. declination or deviation operating with propagation of electric current
Methods comprising measuring the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode are disclosed. Computer readable mediums storing processor executable instructions for performing the method, and systems are also disclosed. The systems comprise a processor, memory and code stored in the memory that when executed cause the processor at least to: receive an output voltage signal, superimpose an excitation signal onto the output voltage signal to produce a superimposed signal, connect the superimposed signal to an electrode in a droplet actuator, suppress the output voltage signal, when detecting an impedance of the electrode, and measure the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode.
G01R 27/16 - Measuring impedance of element or network through which a current is passing from another source, e.g. cable, power line
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
G01N 27/04 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating resistance
The present invention relates to droplet-based particle sorting. According to one embodiment, a method of providing a droplet comprising a single cell is provided, wherein the method includes providing a droplet comprising a suspension of cells on an electrowetting droplet actuator surrounded by an oil medium, dispensing from the droplet to provide a dispensed droplet, determining whether the dispensed droplet on the electrowetting droplet actuator comprises a single cell, and sorting the dispensed droplet on the electrowetting droplet actuator based on the results of the determining step using electrowetting-mediated droplet operations.
The invention is directed to a droplet actuator device and methods for integrating gel electrophoresis analysis with pre or post-analytical sample handling as well as other molecular analysis processes. Using digital microfluidics technology, the droplet actuator device and methods of the invention provide the ability to perform gel electrophoresis and liquid handling operations on a single integrated device. The integrated liquid handling operations may be used to prepare and deliver samples to the electrophoresis gel, capture and subsequently process products of the electrophoresis gel or perform additional assays on the same sample materials which are analyzed by gel electrophoresis. In one embodiment, one or more molecular assays, such as nucleic acid (e.g., DNA) quantification by real-time PCR, and one or more sample processing operations such as sample dilution is performed on a droplet actuator integrated with an electrophoresis gel. In one embodiment, an electrophoresis gel may be integrated on the top substrate of the droplet actuator.
A method of providing a droplet in contact with a magnetically responsive bead and having a reduced quantity of a substance. The method generally includes the steps of (a) providing a droplet actuator comprising: (i) a substrate comprising electrodes arranged for conducting droplet operations on a surface; (ii) a starting droplet comprising: (1) one or more magnetically responsive beads; (2) a starting quantity of the substance; and (3) a starting volume; (b) magnetically immobilizing the one or more magnetically responsive beads at a location which is at a distance from a target droplet splitting zone; (c) conducting one or more droplet operations comprising droplet dividing operations selected to divide the combined droplet to yield a set of droplets comprising: (i) a droplet comprising substantially all of the one or more magnetically responsive beads and having a decreased quantity of the substance relative to the starting concentration; and (ii) a droplet substantially lacking the magnetically responsive beads.
The invention provides a method of circulating magnetically responsive beads within a droplet in a droplet actuator. The invention also provides methods for splitting droplets. The invention, in one embodiment, makes use of a droplet actuator with top and bottom substrates, a plurality of magnetic fields respectively present proximate the top and bottom substrates, wherein at least one of the magnet fields is selectively alterable, and a plurality of droplet operations electrodes positioned along at least one of the top and bottom surfaces. A droplet is positioned between the top and bottom surfaces and at least one of the magnetic fields is selectively altered.
The invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay.
A liquid handling system for supplying liquids to a flow cell (FC). The system may include a droplet actuator cartridge, wherein the droplet actuator and a flow cell are fluidly coupled to, or situated within, a droplet operations gap of the droplet actuator.
The invention is directed to droplet actuator devices and assay methods. The invention includes assay methods of conducting an assay comprising combining a sample with an umbelliferyl derivative, wherein the sample potentially comprises an enzyme capable of cleaving the umbelliferyl derivative and where the umbelliferyl derivative comprises an umbelliferyl core modified with one or more modifying moieties.
C12Q 1/54 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving glucose or galactose
C12Q 1/34 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving hydrolase
C07D 311/18 - Benzo [b] pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted otherwise than in position 3 or 7
C07D 311/20 - Benzo [b] pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 hydrogenated in the hetero ring
C07H 15/203 - Monocyclic carbocyclic rings other than cyclohexane ringsBicyclic carbocyclic ring systems
C12Q 1/25 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving enzymes not classifiable in groups
G01N 33/543 - ImmunoassayBiospecific binding assayMaterials therefor with an insoluble carrier for immobilising immunochemicals
C07D 311/16 - Benzo [b] pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted in position 7
C07D 405/12 - Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
C07D 413/12 - Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
During droplet operations in a droplet actuator, bubbles often form in the filler fluid in the droplet operations gap and interrupt droplet operations. The present invention provides methods and systems for performing droplet operations on a droplet in a droplet actuator comprising maintaining substantially consistent contact between the droplet and an electrical ground while conducting multiple droplet operations on the droplet in the droplet operations gap and/or reducing the accumulation of electrical charges in the droplet operations gap during multiple droplet operations. The methods and systems reduce or eliminate bubble formation in the filler fluid of the droplet operations gap, thereby permitting completion of multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.
The present invention is directed to methods of improving accuracy of droplet metering using at least one on-actuator reservoir as the fluid input. In some embodiments, the on-actuator reservoir that is used for metering droplets includes a loading port, a liquid storage zone, a droplet metering zone, and a droplet dispensing zone. The on-actuator reservoirs are designed to prevent liquid flow-back into the loading port and to prevent liquid from flooding into the droplet operations gap in the dispensing zone.
G01F 23/22 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water
G01F 23/26 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of capacity or inductance of capacitors or inductors arising from the presence of liquid or fluent solid material in the electric or electromagnetic fields
G01F 23/24 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of resistance of resistors due to contact with conductor fluid
34.
DROPLET ACTUATOR TEST CARTRIDGE FOR A MICROFLUIDICS SYSTEM
The present invention is directed to a droplet actuator test cartridge for verifying the operability of a microfluidics system, wherein the test cartridge provides a mechanism for performing diagnostics on a microfluidics system in an automated fashion. In particular, the test cartridge substantially mimics the functions of a droplet actuator and, in so doing, the driving and sensing/receiving functions of a microfluidics system with respect to a droplet actuator can be verified under program control.
G01F 23/24 - Indicating or measuring liquid level or level of fluent solid material, e.g. indicating in terms of volume or indicating by means of an alarm by measuring physical variables, other than linear dimensions, pressure or weight, dependent on the level to be measured, e.g. by difference of heat transfer of steam or water by measuring variations of resistance of resistors due to contact with conductor fluid
The present invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay.
The invention provides droplet actuators and droplet actuator cassettes including reagent storage capabilities, as well as methods of making and using the droplet actuators and cassettes. The invention also provides continuous flow channel elements and techniques for using electrodes to manipulate droplets in flowing streams. The invention also discloses methods of separating compounds on a droplet actuator. Various other aspects of the invention are also disclosed.
The present invention provides methods for on-actuator temperature measurement and temperature control, including where one or more of the temperature sensors are combined with one or more heaters that are formed of wiring traces and/or providing heaters designed for one-to- one correspondence to the temperature sensors to form temperature sensor-heater pairs. The present invention also provides methods for on-actuator temperature measurement and temperature control in which the temperature sensors comprise a connection comprising a plurality of terminals by which an amount of current can be applied and then a voltage measured, wherein the voltage that is measured across the temperature sensors can be accurately correlated to a temperature.
B05B 15/00 - Details of spraying plant or spraying apparatus not otherwise provided forAccessories
B05B 1/24 - Nozzles, spray heads or other outlets, with or without auxiliary devices such as valves, heating means incorporating means for heating the liquid or other fluent material, e.g. electrically
G01K 13/00 - Thermometers specially adapted for specific purposes
G01K 7/00 - Measuring temperature based on the use of electric or magnetic elements directly sensitive to heat
The invention provides a method of redistributing magnetically responsive beads in a droplet. The method may include conducting on a droplet operations surface one or more droplet operations using the droplet without removing the magnetically responsive beads from the region of the magnetic field. The droplet operations may in some cases be electrode-mediated. The droplet operations may redistribute and/or circulate the magnetically responsive beads within the droplet. In some cases, the droplet may include a sample droplet may include a target analyte. The redistributing of the magnetically responsive beads may cause target analyte to bind to the magnetically responsive beads. In some cases, the droplet may include unbound substances in a wash buffer. The redistributing of the magnetically responsive beads causes unbound substances to be freed from interstices of an aggregated set or subset of the magnetically responsive beads.
The invention provides a droplet actuator designed to provide reliable electrical connections to droplets to reduce or eliminate gas bubble formation during droplet operations. The invention also provides methods and systems for reducing or eliminating the formation of bubbles in a droplet during droplet operations.
The invention provides a droplet actuator designed for performing electroporation on cells in droplets. The invention also provides method and systems for performing electroporation on cells in droplets on a droplet actuator.
A method, circuit and apparatus for detecting capacitance on a droplet actuator, inter alia, for determining the presence, partial presence or absence of a droplet at an electrode on a droplet actuator by: (a) providing a droplet actuator comprising: (i) a substrate comprising electrodes arranged on the substrate for conducting droplet operations on a surface of the substrate; (ii) a capacitance detection circuit for detecting capacitance at the droplet operations surface at one or more of the electrodes; (b) detecting capacitance at the droplet operations surface at one or more of the electrodes; and (c) determining from the capacitance the presence, partial presence or absence of a droplet at the droplet operations surface at the electrode.
G01R 27/26 - Measuring inductance or capacitanceMeasuring quality factor, e.g. by using the resonance methodMeasuring loss factorMeasuring dielectric constants
G01N 25/08 - Investigating or analysing materials by the use of thermal means by investigating changes of state or changes of phaseInvestigating or analysing materials by the use of thermal means by investigating sintering of boiling point
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
G01N 27/22 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance
Aspects of embodiments may include methods for automated enzymatic detection of glucose-6-phosphate dehydrogenase (G6PD) activity. Aspects of embodiments may include methods for enzymatic detection of G6PD activity in droplets in oil. Aspects of embodiments may include a system including a droplet actuator. Aspects of embodiments may include a treatment method.
The invention provides pyrosequencing-based methods of analyzing and synthesizing DNA, including methods of DNA error correction, determining DNA size distribution, screening for nucleotide repeat disorders such as fragile X syndrome, determining size distribution and bias in a DNA library, and determining pyrosequencing read length. The methods include on-bench protocols as well as droplet-based protocols that may be conducted on a droplet actuator.
The invention provides a method of dispersing or circulating magnetically responsive beads within a droplet in a droplet actuator. The invention, in one embodiment, makes use of a droplet actuator with a plurality of droplet operations electrodes configured to transport the droplet, and a magnetic field present at a portion of the plurality of droplet operations electrodes. A bead-containing droplet is provided on the droplet actuator in the presence of the uniform magnetic field. Beads are circulated in the droplet during incubation by conducting droplet operations on the droplet within a uniform region of the magnate field. Other embodiments are also provided.
Droplet actuators and techniques for droplet-based assays are provided. A method of conducting an assay comprises: incubating a droplet in oil, the droplet comprising an umbelliferone substrate, a sample potentially comprising an enzyme which cleaves the umbelliferone substrate, and a zwitterionic surfactant; and detecting a signal emitted from the droplet.
The invention provides a droplet actuator. The droplet actuator may include a base substrate and a top substrate separated to form a gap. The base substrate may include electrodes configured for conducting droplet operations in the gap; and the top substrate may include a glass substrate portion coupled to a non-glass portion, where the non-glass portion may include one or more openings establishing a fluid path extending from an exterior of the droplet actuator and into the gap. The invention also provides related methods of manufacturing the droplet actuator, methods of using the droplet actuator, and methods of loading the droplet actuator.
Methods comprising measuring the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode are disclosed. Computer readable mediums storing processor executable instructions for performing the method, and systems are also disclosed. The systems comprise a processor, memory and code stored in the memory that when executed cause the processor at least to: receive an output voltage signal, superimpose an excitation signal onto the output voltage signal to produce a superimposed signal, connect the superimposed signal to an electrode in a droplet actuator, suppress the output voltage signal, when detecting an impedance of the electrode, and measure the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode.
The invention provides methods for gene synthesis and error correction on a droplet actuator. In one embodiment, the invention provides a method for performing polymerase-mediated gene synthesis on a droplet actuator. The polymerase-mediated gene synthesis method uses oligonucleotide hybridization and PCR cycling to generate a pool of synthesized DNA strands, which may be subsequently enriched for the correct gene sequence.
The present invention is directed to mechanisms for and methods of loading a droplet actuator with filler fluid. The mechanisms and methods are useful for loading filler fluid in a droplet actuator in a manner that is rapid enough for practical use while at the same time sufficiently slow to reduce, preferably entirely avoid, the trapping of air in the droplet operations gap during the loading process.
The invention is directed to an assay for acid β-galactosidase activity. The invention may include combining in oil a sample droplet with a 4-methylumbelliferyl-B-galactose to yield a reaction droplet; splitting the reaction droplet to yield a first daughter droplet and a second daughter droplet; combining the first daughter droplet with a stop buffer droplet to yield a first stopped reaction droplet; incubating the second daughter droplet; combining the second daughter droplet with a stop buffer droplet to yield a second stopped reaction droplet; and measuring 4-methylumbelliferone released in the first and second stopped reaction droplets.
C12Q 1/54 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving glucose or galactose
C07D 311/18 - Benzo [b] pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted otherwise than in position 3 or 7
C07D 311/20 - Benzo [b] pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 hydrogenated in the hetero ring
C07H 15/203 - Monocyclic carbocyclic rings other than cyclohexane ringsBicyclic carbocyclic ring systems
C12Q 1/25 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving enzymes not classifiable in groups
C12Q 1/34 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving hydrolase
A method for conducting medium-chain acyl-CoA dehydrogenase (MCAD) and very- long-chain acyl-CoA dehydrogenase (VCAD) enzymatic activity assays is provided. The method may include, but is not limited to, preparing a sample; preparing an enzyme- specific substrate/reagent mixture; mixing an aliquot of the prepared sample with an aliquot of the enzyme-specific substrate/reagent mixture; reading absorbance in the range of about 600 nm; incubating the prepared sample and enzyme-specific substrate/reagent mixture; and reading absorbance in the range of about 600 nm at various time intervals.
C12Q 1/32 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving oxidoreductase involving dehydrogenase
C12Q 1/26 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving oxidoreductase
G01N 33/48 - Biological material, e.g. blood, urineHaemocytometers
52.
DIGITAL MICROFLUIDICS CARTRIDGE AND SYSTEM FOR OPERATING A FLOW CELL
A liquid handling system for supplying liquids to a flow cell (FC). The system may include a droplet actuator cartridge, wherein the droplet actuator and a flow cell are fluidly coupled to, or situated within, a droplet operations gap of the droplet actuator.
C12M 1/00 - Apparatus for enzymology or microbiology
C12M 1/34 - Measuring or testing with condition measuring or sensing means, e.g. colony counters
C12M 1/36 - Apparatus for enzymology or microbiology including condition or time responsive control, e.g. automatically controlled fermentors
G01N 33/48 - Biological material, e.g. blood, urineHaemocytometers
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
The present invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay.
The invention provides droplet actuators and droplet actuator techniques. Among other things, the droplet actuators and methods are useful for manipulating beads on a droplet actuator, such as conducting droplet operations using bead-containing droplets on a droplet actuator. For example, beads may be manipulated on a droplet actuator in the context of executing a sample preparation protocol and/or an assay protocol. An output of the methods of the invention may be beads prepared for execution of an assay protocol. Another output of the methods of the invention may be results of an assay protocol executed using beads. Among the methods described herein are methods of concentrating beads in droplets, methods of washing beads, methods of suspending beads, methods of separating beads, methods of localizing beads within a droplet, methods of forming emulsions in which droplets include beads, methods of loading beads into a droplet operations gap of a droplet actuator, methods of organizing beads in a monolayer, and methods of capturing, trapping or restraining beads.
During droplet operations in a droplet actuator, bubbles often form in the filler fluid in the droplet operations gap and interrupt droplet operations. The present invention provides methods and systems for performing droplet operations on a droplet in a droplet actuator comprising maintaining substantially consistent contact between the droplet and an electrical ground while conducting multiple droplet operations on the droplet in the droplet operations gap and/or reducing the accumulation of electrical charges in the droplet operations gap during multiple droplet operations. The methods and systems reduce or eliminate bubble formation in the filler fluid of the droplet operations gap, thereby permitting completion of multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.
During droplet operations in a droplet actuator, bubbles often form in the filler fluid in the droplet operations gap and interrupt droplet operations. The present invention provides methods and systems for performing droplet operations on a droplet in a droplet actuator comprising maintaining substantially consistent contact between the droplet and an electrical ground while conducting multiple droplet operations on the droplet in the droplet operations gap and/or reducing the accumulation of electrical charges in the droplet operations gap during multiple droplet operations. The methods and systems reduce or eliminate bubble formation in the filler fluid of the droplet operations gap, thereby permitting completion of multiple droplet operations without interruption by bubble formation in the filler fluid in the droplet operations gap.
Droplet actuators that include molecular barrier coatings are provided. The molecular barrier coating may be provided atop the conductive layer of the top substrate, atop the droplet operations electrodes of the bottom substrate, or both. Where the conductive layer of the top substrate and/or the droplet operations electrodes of the bottom substrate are formed of an electrically conductive organic polymer, such as poly(3,4-ethylenedioxythiophene) poly(styrenesulfonate) (PEDOT:PSS), the molecular barrier coating helps to immobilize the contents of the PEDOT:PSS layer. Further, the molecular barrier coating reduces, preferably entirely eliminates, moisture from seeping into the electrically conducting organic polymer. Methods of conducting droplet operations using the disclosed droplet actuators are also provided.
The invention relates to methods for enzymatic detection of biotinidase activity. In certain aspects, the invention provides methods for bench-based enzymatic detection of biotinidase activity. In other aspects, the invention provides methods for bench-based enzymatic detection in droplets in oil. The droplet-based methods may be performed in an automated manner using digital microfluidics technology on a droplet actuator device. The enzymatic assays for biotinidase activity may be used for newborn testing for biotinidase deficiency, and may be combined with other droplet-based enzymatic assays in a panel of tests for newborn testing.
C12Q 1/25 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving enzymes not classifiable in groups
A method of detecting a target analyte in a sample, including executing electrowetting-mediated droplet operations and thereby: combining one or more immunoassay reagent droplets comprising magnetically-responsive beads having affinity for the target analyte with one or more sample droplets potentially comprising the target analyte to yield a first combined droplet; beginning with the combined droplet, effecting a droplet-based washing protocol to wash the magnetically-responsive beads to yield a first washed droplet comprising the washed magnetically responsive beads; and combining the droplet comprising the washed magnetically responsive beads with a droplet comprising a reporter molecule having affinity for target analyte captured on the magnetically-responsive beads to yield a second combined droplet; beginning with the second combined droplet, effecting a droplet-based washing protocol to wash the magnetically-responsive beads to yield a second washed droplet comprising the washed magnetically responsive beads; detecting a signal from the second washed droplet which corresponds to the presence, absence and/or quantity of the analyte in the sample.
G01N 33/543 - ImmunoassayBiospecific binding assayMaterials therefor with an insoluble carrier for immobilising immunochemicals
G01N 27/72 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating magnetic variables
G01N 27/74 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating magnetic variables of fluids
60.
Method of conducting a droplet based enzymatic assay
A method is provided for conducting a droplet-based enzymatic assay, e.g., for diagnostic purposes. On a droplet actuator, a droplet comprising an enzyme of interest is provided along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. Droplet operations are executed to combine the enzyme and substrate droplets on the droplet actuator, thereby yielding an assay droplet on the droplet actuator. Detecting modification of the substrate by the enzyme in the assay droplet occurs on the droplet actuator. Modified substrate preparations for conducting such enzymatic assays are also provided.
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions
The invention relates to a droplet actuator device and methods for integrated sample preparation and analysis of a biological sample. A droplet actuator device is provided for conducting droplet operations. The droplet actuator device may include a bottom substrate and a top substrate separated from each other to form a gap therebetween; an arrangement of droplet operations electrodes arranged on one or both of the bottom and/or top substrates for conducting droplet operations thereon; a reagent storage layer comprising one or more compartments bound to the top substrate; and one or more openings arranged to provide a fluidic path from the one or more compartments into the gap, upon breach of a breachable seal separating the one or more compartments and openings.
The invention provides droplet actuator assemblies and systems and methods of manufacturing the droplet actuator assemblies. In certain embodiments, two-piece enclosures are used to form a droplet actuator assembly that houses a droplet operations substrate. In certain other embodiments, one-piece enclosures are used to form a droplet actuator assembly that houses a droplet operations substrate. In the plastic injection molding process for forming substrates of the droplet actuator assemblies of the present invention may utilize insert molding (or overmolding) processes for forming a gasket in at least one substrate, thereby avoiding the need for providing and installing a separate gasket component. Further, the droplet actuator assemblies may include features that allow ultrasonic welding processes to be used for bonding substrates together. The manufacturing systems of the present invention for fabricating the droplet actuator assemblies may utilize continuous flow reel-to-reel manufacturing processes.
The present invention relates to amplification oligonucleotides (primers) and detection probes for the detection of human immunodeficiency virus type-1 (HIV-1) in a test sample. The primers and probes may be used in an assay for the detection and/or quantitation of HIV-1 nucleic acid. In various embodiments, the primers and detection probes may be used for real-time reverse transcription polymerase chain reaction (RT-PCR) assays for detection of HIV-1 nucleic acid. The detection probes may, for example, be fluorescently-labeled TaqMan® probes. In one embodiment, amplification primers and probes may be specific for the pol region of the HIV-1 genome. In another embodiment, the amplification primers and probes may be specific for the env region of the HIV-1 genome. "Cocktails" of multiple amplification primers and/or probes may be used to achieve maximal coverage of different HIV-1 subtypes.
The present invention relates to methods of sample preparation on a droplet actuator. The invention further relates to methods of plasma sample preparation on a droplet actuator by immunocapture. The invention also relates to methods of preparing plasma that contains HIV from whole blood without the need for filtration or centrifugation. Additionally, the invention relates to methods of sample preparation that use hydrophobic chromatography for purifying or removing unwanted materials from the sample. The invention further relates to a droplet actuator and methods of loading solutions of large volume into a droplet actuator under controlled conditions. The invention also relates to methods of processing cells on a droplet actuator.
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 1/10 - Devices for withdrawing samples in the liquid or fluent state
Aspects of embodiments may include methods for automated enzymatic detection of glucose-6-phosphate dehydrogenase (G6PD) activity. Aspects of embodiments may include methods for enzymatic detection of G6PD activity in droplets in oil. Aspects of embodiments may include a system including a droplet actuator. Aspects of embodiments may include a treatment method.
C12Q 1/26 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving oxidoreductase
G01N 33/68 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing involving proteins, peptides or amino acids
G01N 33/52 - Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper
The invention is directed to droplet actuator devices and assay methods. The invention includes assay methods of conducting an assay comprising combining a sample with an umbelliferyl derivative, wherein the sample potentially comprises an enzyme capable of cleaving the umbelliferyl derivative and where the umbelliferyl derivative comprises an umbelliferyl core modified with one or more modifying moieties.
Aspects of embodiments of the invention relate to a simulator including a visual display capable of outputting to a user a display one or more effects of a command series selected and a system including a droplet microactuator electronically coupled to and controlled by a processor capable of executing instructions, the droplet microactuator including a substrate comprising electrodes for conducting droplet operations. Further aspects of embodiments of the invention relate to a droplet operations troubleshooting apparatus. Other aspects of embodiments of the invention relate to a computer implemented method of displaying simulated microactuator droplets.
The present invention relates to apparatuses for delivering one or more volumes of fluids to one or more microfluidics devices. The invention further relates to methods for delivering one or more volumes of fluids to one or more microfluidics devices. Additionally, the invention relates to systems that include apparatuses for delivering one or more volumes of fluids to one or more microfluidics devices.
G01N 35/10 - Devices for transferring samples to, in, or from, the analysis apparatus, e.g. suction devices, injection 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
69.
DROPLET ACTUATOR WITH IMPROVED WASTE DISPOSAL CAPABILITY
Methods, devices and systems for transporting a droplet along a droplet operations gap of a droplet actuator into contact with absorbent material to dispose of the droplet from the gap or from a region of the gap.
G01N 27/26 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variablesInvestigating or analysing materials by the use of electric, electrochemical, or magnetic means by using electrolysis or electrophoresis
G01N 15/02 - Investigating particle size or size distribution
G01N 27/30 - Electrodes, e.g. test electrodesHalf-cells
Droplet actuator apparatus and system are provided. An apparatus comprises: a microfluidics system having an insertion slot for insertion of a droplet actuator; a bottom plate against which the droplet actuator would slide when inserting the droplet actuator into the insertion slot; and means for forcing a substrate in the droplet actuator parallel to the bottom plate, wherein warpage in the substrate is corrected.
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 1/10 - Devices for withdrawing samples in the liquid or fluent state
G01N 27/02 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance
71.
DROPLET ACTUATORS AND TECHNIQUES FOR DROPLET-BASED ASSAYS
Droplet actuators and techniques for droplet-based assays are provided. A method of conducting an assay comprises: incubating a droplet in oil, the droplet comprising an umbelliferone substrate, a sample potentially comprising an enzyme which cleaves the umbelliferone substrate, and a zwitterionic surfactant; and detecting a signal emitted from the droplet.
A method of providing a droplet comprising one or more reagents, the method comprising, depositing a first aqueous droplet comprising the one or more reagents on a surface; drying the droplet to yield a dried composition on the surface comprising the one or more reagents; covering the dried composition with oil; and causing a second aqueous droplet in the oil to contact the dried composition and thereby resuspend one or more reagents.
Methods comprising measuring the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode are disclosed. Computer readable mediums storing processor executable instructions for performing the method, and systems are also disclosed. The systems comprise a processor, memory and code stored in the memory that when executed cause the processor at least to: receive an output voltage signal, superimpose an excitation signal onto the output voltage signal to produce a superimposed signal, connect the superimposed signal to an electrode in a droplet actuator, suppress the output voltage signal, when detecting an impedance of the electrode, and measure the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode.
A method of preparing a sample for conducting an assay, the method including: providing an input sample including glycoproteins; capturing glycoproteins from the input sample on a solid support; and washing the sample support to remove unbound portions of the input sample.
Methods comprising measuring the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode are disclosed. Computer readable mediums storing processor executable instructions for performing the method, and systems are also disclosed. The systems comprise a processor, memory and code stored in the memory that when executed cause the processor at least to: receive an output voltage signal, superimpose an excitation signal onto the output voltage signal to produce a superimposed signal, connect the superimposed signal to an electrode in a droplet actuator, suppress the output voltage signal, when detecting an impedance of the electrode, and measure the impedance of the electrode produced by the excitation signal, wherein the impedance indicates presence of liquid at the electrode.
A method of mixing a droplet, the method comprising providing a droplet on a surface, forming the droplet into a first "U" shape having a bottom region and two terminal ends, and simultaneously merging the terminal ends and splitting the droplet at the bottom region to form a second "U" shape which is substantially opposite the first "U" shape.
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
G01N 21/00 - Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
B01F 3/08 - Mixing, e.g. dispersing, emulsifying, according to the phases to be mixed liquids with liquids; Emulsifying
The invention is directed to droplet actuator devices and assay methods. The method may include immobilization of the enzymatic substrate including forming an inclusion complex with the substrate within an aqueous environment in contact with an oil.
The embodiments described herein provide methods of measuring capacitance, detecting a droplet at a position, determining a thickness of an oil film and determining temperature in a droplet actuator. Specifically, the capacitance detection method may be used as a real-time verification tool in order to detect the absence, presence, and/or partial presence of a droplet at an electrode, analyze droplet properties, measure droplet size or volume, optimize the speed of droplet operation and detect air bubbles.
G01R 27/26 - Measuring inductance or capacitanceMeasuring quality factor, e.g. by using the resonance methodMeasuring loss factorMeasuring dielectric constants
The present invention is directed to droplet actuator systems, devices, and methods. In one embodiment, a microfluidic article of manufacture is provided. The microfluidic article of manufacture includes a first substrate; a second substrate separated from the first substrate to form a droplet operations gap; gap height setting spacers associated with the first and/or second substrate or situated between the first and second substrates; a spring forcing the second substrate against the gap height setting spacers, thereby establishing a substantially uniform gap height between the first and second substrates; and electrodes associated with the first and/or second substrate and configured to conduct droplet operations in the droplet operations gap.
B01J 19/00 - Chemical, physical or physico-chemical processes in generalTheir relevant apparatus
G01N 27/26 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variablesInvestigating or analysing materials by the use of electric, electrochemical, or magnetic means by using electrolysis or electrophoresis
The invention provides droplet actuator assemblies and systems and methods of manufacturing the droplet actuator assemblies. In certain embodiments, two-piece enclosures are used to form a droplet actuator assembly that houses a droplet operations substrate. In certain other embodiments, one-piece enclosures are used to form a droplet actuator assembly that houses a droplet operations substrate. In the plastic injection molding process for forming substrates of the droplet actuator assemblies of the present invention may utilize insert molding (or overmolding) processes for forming a gasket in at least one substrate, thereby avoiding the need for providing and installing a separate gasket component. Further, the droplet actuator assemblies may include features that allow ultrasonic welding processes to be used for bonding substrates together. The manufacturing systems of the present invention for fabricating the droplet actuator assemblies may utilize continuous flow reel-to-reel manufacturing processes.
The invention relates to a droplet actuator for conducting droplet operations. The actuator includes a bottom substrate and a top substrate separated from the bottom substrate to form a gap. An arrangement of droplet operations electrodes may be located on a surface of the bottom substrate and/or top substrate. Optionally, a sample reservoir may hold a quantity of a sample fluid containing cells. A disruption device which can take various forms is used to lyse the cells in the sample or in a sample droplet to thereby conduct operations on samples having lysed cells therein.
The invention relates to certain novel approaches to reducing or eliminating the movement of contaminants from one droplet to another on a droplet actuator via liquid filler fluid. In one application, droplet actuators are used to conduct genetic analysis using polymerase chain reaction (PCR) techniques. The invention addresses the need for improved methods of performing PCR on a droplet actuator that provide for optimum amplification and detection of a sample target.
The invention relates to a droplet actuator device and methods for integrated sample preparation and analysis of a biological sample. A droplet actuator device is provided for conducting droplet operations. The droplet actuator device may include a bottom substrate and a top substrate separated from each other to form a gap therebetween; an arrangement of droplet operations electrodes arranged on one or both of the bottom and/or top substrates for conducting droplet operations thereon; a reagent storage layer comprising one or more compartments bound to the top substrate; and one or more openings arranged to provide a fluidic path from the one or more compartments into the gap, upon breach of a breachable seal separating the one or more compartments and openings.
A method of preparing a nucleic acid library in droplets in contact with oil, including: (a) blunt-ending nucleic acid fragments in a droplet in the oil to yield blunt-ended nucleic acid fragments; (b) phosphorylating the blunt-ended nucleic acid fragments in a droplet in the oil to yield phosphorylated nucleic acid fragments; coupling A-tails to the phosphorylated nucleic acid fragments in a droplet in the oil to yield A-tailed nucleic acid fragments; and (d) coupling nucleic acid adapters to the A-tailed nucleic acid fragments in a droplet in the oil to yield the nucleic acid library comprising adapter-ligated nucleic acid fragments.
C12N 15/10 - Processes for the isolation, preparation or purification of DNA or RNA
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
C40B 40/06 - Libraries containing nucleotides or polynucleotides, or derivatives thereof
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
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
86.
Methods, systems, and products for conducting droplet operations
The present invention provides modified droplet actuator systems, software, and software-executed methods for use in droplet actuator operation and droplet actuator systems that are configured and programmed to execute such software. An aspect of the software components of the invention is an interface description file for each hardware component of a microfluidics system that allows hardware components to be changed without modifying the program for performing droplet operations protocols. Another aspect of the software components of the invention is the establishment of electrode-to-electrode relationships and other aspects of droplet actuator configurations, which may be used when programming droplet operations protocols. Another aspect of the software components of the invention is a physical design library of predefined electrode elements that may be used by a droplet actuator designer when constructing a layout of electrodes. Another aspect of the software components of the invention is a droplet actuator description file that contains the physical and electrical description of the droplet actuator. Another aspect of the software components of the invention is a router component for determining routes of droplet operations in a droplet actuator. Another aspect of the software components of the invention is the use of tri-state vectors for programming sequences in a droplet actuator. Still other aspects are provided.
The present invention relates to bead incubating and washing on a droplet actuator. Methods for incubating magnetically responsive beads that are labeled with primary antibody, a sample (i.e., analyte), and secondary reporter antibodies on a magnet, on and off a magnet, and completely off a magnet are provided. Also provided are methods for washing magnetically responsive beads using shape-assisted merging of droplets. Also provided are methods for shape-mediated splitting, transporting, and dispensing of a sample droplet that contains magnetically responsive beads. The apparatuses and methods of the invention provide for rapid time to result and optimum detection of an analyte in an immunoassay.
The invention is directed to droplet actuator devices and assay methods for multiplexed newborn testing for metabolic disorders. The methods include, among other things, droplet-based enzymatic assays and immunoassays for testing for metabolic disorders. The invention includes methods and devices for conducting multiple assays for different metabolic disorders on a single droplet actuator, as well as multiple assays for the same metabolic disorder using samples from different subjects and/or multiple samples from the same subject on a single droplet actuator. In various embodiments, the invention includes methods for conducting enzymatic activity assays and/or immunoassays in a single fresh blood and/or plasma samples and dried blood and/or plasma samples.
C12Q 1/34 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving hydrolase
C12Q 1/25 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving enzymes not classifiable in groups
C12M 1/34 - Measuring or testing with condition measuring or sensing means, e.g. colony counters
G01N 33/52 - Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper
C07D 311/06 - Benzo [b] pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2
C07D 311/18 - Benzo [b] pyrans, not hydrogenated in the carbocyclic ring with oxygen or sulfur atoms directly attached in position 2 not hydrogenated in the hetero ring substituted otherwise than in position 3 or 7
89.
Enzymatic assays using umbelliferone substrates with cyclodextrins in droplets in oil
The invention is directed to droplet actuator devices and assay methods. The method may include, among other things, a method of conducting enzymatic assays using umbelliferone substrates with cyclodextrins in droplets in oil, the method including incubating a droplet in oil, the droplet preferably comprising an umbelliferone substrate, a sample, and a cyclodextrin compound. The methods may further include a method of substantially eliminating cross-contamination between droplets in enzymatic containing substrate-based bioassays. The method may include immobilization of the enzymatic substrate including forming an inclusion complex with the substrate for stabilizing the substrate within an aqueous environment, wherein the inclusion complex may be formed using cyclodextrins. In yet another embodiment, the invention may provide a method of enhancing hydrolysis of enzymatic substrates. The methods may further include a method of forming an inclusion complex with the substrate for stabilizing the substrate within an aqueous environment, wherein the substrate may be a 4-MU- or HMU containing substrate. The inclusion complex may be formed using cyclodextrin.
The invention is directed to a droplet actuator device and methods for integrating gel electrophoresis analysis with pre or post-analytical sample handling as well as other molecular analysis processes. Using digital microfluidics technology, the droplet actuator device and methods of the invention provide the ability to perform gel electrophoresis and liquid handling operations on a single integrated device. The integrated liquid handling operations may be used to prepare and deliver samples to the electrophoresis gel, capture and subsequently process products of the electrophoresis gel or perform additional assays on the same sample materials which are analyzed by gel electrophoresis. In one embodiment, one or more molecular assays, such as nucleic acid (e.g., DNA) quantification by real-time PCR, and one or more sample processing operations such as sample dilution is performed on a droplet actuator integrated with an electrophoresis gel. In one embodiment, an electrophoresis gel may be integrated on the top substrate of the droplet actuator.
C12Q 1/68 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions involving nucleic acids
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
G01N 33/48 - Biological material, e.g. blood, urineHaemocytometers
G01N 35/00 - Automatic analysis not limited to methods or materials provided for in any single one of groups Handling materials therefor
Methods of concentrating beads in a droplet and/or loading beads on a fluidic device are provided, including among other things, a method of concentrating beads in a droplet, the method comprising: (a) providing a droplet actuator comprising: (i) an interior droplet operations volume; and (ii) a reservoir exterior to the interior volume; (iii) a droplet established in a liquid path extending from the reservoir into the interior volume; (b) providing magnetically responsive beads in the portion of the droplet which is in the reservoir; (c) magnetically attracting the magnetically responsive beads through the liquid path into the portion of the droplet which is in the interior volume; and (d) forming a droplet comprising one or more of the magnetically responsive beads in the interior volume.
A droplet actuator comprising: (a) a base substrate comprising electrodes configured for conducting droplet operations on a droplet operations surface thereof; (b) a droplet comprising one or more beads situated on the droplet operations surface; (c) a barrier arranged in relation to the droplet and the electrodes such that a droplet may be transported away from the beads using one or more droplet operations mediated by one or more of the electrodes while transport of the beads is restrained by a barrier. Related methods and kits are also provided.
The invention is directed to certain droplet actuated molecular techniques. In one embodiment, the invention provides droplet actuator methods for detection of single nucleotide polymorphisms (SNPs) in a DNA sequence using digital microfluidics, including droplet actuator-based sample preparation and SNP analysis. In another embodiment, the invention provides droplet actuator devices and methods for providing integrated sample preparation and multiplexed detection of an infectious agent, such as HIV. In yet another embodiment, the invention provides droplet actuator devices and techniques for PCR amplification and detection of specific nucleic acid sequences using digital microfluidics, including droplet actuator-based sample preparation and target nucleic acid analysis. In yet another embodiment the invention provides methods for performing hot-start PCR on a droplet actuator. In yet another embodiment, the method of the invention combines PCR amplification with pyrosequencing to investigate specific sequences.
The invention provides droplet actuator devices for facilitating certain droplet actuated molecular techniques. In one embodiment, the invention provides droplet actuators that facilitate droplet operations in three dimensions. In another embodiment, the invention provides for cost-effective production of droplet actuators. In another embodiment, the invention provides for replacing one or more components of a droplet actuator. In another embodiment, the invention provides droplet actuators using printed conductive inks to form electrodes and/or ground planes. In another embodiment, the invention provides a magnetic clamping fixture for assembling droplet actuators. In another embodiment, the invention provides simple, low cost power sources for use in combination with microfluidic systems. In another embodiment, the invention provides an immunoassay multiplexing platform that uses digital microfluidics and real-time imaging of flash-based chemiluminescent signals. In still another embodiment, the invention provides droplet actuator devices that are fabricated using a plasma treatment process to raise substrate surface energy.
A method of amplifying a nucleic acid, the method comprising cycling an amplification-ready droplet through a temperature gradient to locations within the temperature gradient suitable for effecting steps in an amplification reaction.
The invention provides a droplet actuator comprising a base substrate and a top substrate separated to form a gap, wherein: (a) the base substrate comprises electrodes configured for conducting droplet operations in the gap; and (b) the top substrate comprises a first portion coupled to second portion, where the second portion comprises one or more openings establishing a fluid path extending from an exterior of the droplet actuator and into the gap. The invention also provides related methods of manufacturing the droplet actuator, methods of using the droplet actuator, and methods of loading the droplet actuator.
G01N 27/26 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variablesInvestigating or analysing materials by the use of electric, electrochemical, or magnetic means by using electrolysis or electrophoresis
B01L 3/00 - Containers or dishes for laboratory use, e.g. laboratory glasswareDroppers
The invention provides a method of dispersing or circulating magnetically responsive beads within a droplet in a droplet actuator. The invention, in one embodiment, makes use of a droplet actuator with a plurality of droplet operations electrodes configured to transport the droplet, and a magnetic field present at a portion of the plurality of droplet operations electrodes. A bead-containing droplet is provided on the droplet actuator in the presence of the uniform magnetic field. Beads are circulated in the droplet during incubation by conducting droplet operations on the droplet within a uniform region of the magnetic field wherein droplet operations do not allow magnetically responsive beads to be introduced into a region of the magnetic field which is sufficiently non-uniform to cause bead clumping resulting in a more homogenous distribution of the beads in the droplet. Other embodiments are also provided.
A method, circuit and apparatus for detecting capacitance on a droplet actuator, inter alia, for determining the presence, partial presence or absence of a droplet at an electrode on a droplet actuator by: (a) providing a droplet actuator comprising: (i) a substrate comprising electrodes arranged on the substrate for conducting droplet operations on a surface of the substrate; (ii) a capacitance detection circuit for detecting capacitance at the droplet operations surface at one or more of the electrodes; (b) detecting capacitance at the droplet operations surface at one or more of the electrodes; and (c) determining from the capacitance the presence, partial presence or absence of a droplet at the droplet operations surface at the electrode.
G01R 27/26 - Measuring inductance or capacitanceMeasuring quality factor, e.g. by using the resonance methodMeasuring loss factorMeasuring dielectric constants
G01N 27/22 - Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating impedance by investigating capacitance
99.
NUCLEIC ACID AMPLIFICATION AND SEQUENCING ON A DROPLET ACTUATOR
The invention provides a droplet actuator device, as well as systems, methods and devices making use of the droplet actuator device. The droplet actuator device may include a substrate having electrodes arranged for conducting one or more droplet operations. The droplet actuator device may include a substrate having a reactor path with a wash region associated with a magnet for immobilizing beads during bead washing operations. The droplet actuator device may include nucleotide base reservoirs and dedicated nucleotide base electrode paths arranged for transporting nucleotide base droplets from nucleotide base reservoirs to the reactor path. The droplet actuator device may include one or more wash buffer reservoirs associated with electrode paths arranged for transporting wash buffer droplets from wash buffer reservoirs to the reactor path. The droplet actuator device may include one or more sample reservoirs and sample paths arranged for transporting sample droplets from the one or more sample reservoirs to the reactor path. The droplet actuator device may include one or more enzyme reservoirs and dedicated enzyme electrode paths arranged for transporting enzyme droplets from the one or more enzyme reservoirs to a detection electrode.
A method of conducting a droplet-based enzymatic assay is provided. The method generally makes use of a droplet actuator. A droplet comprising an enzyme of interest is provided on the droplet actuator along with a droplet comprising a substrate which is potentially modified in the presence of the enzyme. The method involves executing droplet operations on the droplet actuator to combine the droplets, thereby yielding an assay droplet, and detecting modification of the substrate by the enzyme in the assay droplet on the droplet actuator. The enzyme of interest may, for example, be a potentially mutated or improperly folded enzyme exhibiting altered enzyme activity as compared to a corresponding normal enzyme.
C12Q 1/00 - Measuring or testing processes involving enzymes, nucleic acids or microorganismsCompositions thereforProcesses of preparing such compositions
patents
