Abstract: System, including methods, apparatus, compositions, and kits, for the mixing of small volumes of fluid by coalescence of multiple emulsions.

Abstract: The invention provides a system for high-throughput multiplex analysis of target samples. A sample and reagent delivery unit is operatively connected to a thermal cycler for amplification of target nucleic acids. Microspheres are hybridized to the resulting amplicons in the thermal cycler. A flow cytometer is operatively connected to the thermal cycler or optionally a bead trap for washing the microspheres.

Abstract: Method of performing a multiplexed digital assay. The method may include (1) partitioning a sample containing at least two distinct targets into droplets; (2) amplifying the at least two distinct targets within the droplets; (3) detecting light from the droplets indicative of the presence or absence of each distinct target in each droplet; and (4) calculating a level of each distinct target based on the detected light.

Abstract: A system, including method and apparatus, for forming an array of emulsions. The system may include a plate providing an array of emulsion production units each configured to produce a separate emulsion and each including a set of wells interconnected by channels that intersect to form a site of droplet generation. Each set of wells, in turn, may include (1) at least one first input well to receive a continuous phase, (2) a second input well to receive a dispersed phase, and (3) an output well configured to receive from the site of droplet generation an emulsion of droplets of the dispersed phase disposed in the continuous phase.

Abstract: System, including methods, apparatus, and kits, for forming emulsions. The system may include an instrument and a microfluidic chip received by the instrument. The instrument may apply pressure to prospective emulsion phases held by the chip, to drive formation and collection of emulsions in the chip. In some embodiments, the instrument may stop applying pressure to the chip when a change in pressure meeting a predefined condition is detected by the instrument. The change may indicate that an endpoint of droplet generation has been reached.

Abstract: System, including methods and compositions, for making and using emulsions that include a silicone oil and a silicone surfactant. The emulsions may include aqueous droplets disposed in a continuous phase that includes a silicone oil and a silicone surfactant. The aqueous droplets may contain an analyte, optionally at partial occupancy, and/or a luminescent (e.g., photoluminescent) reporter. An assay of the analyte may be performed with the droplets. In some cases, signals may be detected from the droplets, and a characteristic of the analyte, such as an analyte level or activity, may be determined based on the signals.

Abstract: System, including methods and apparatus, for detection of spaced droplets.

Abstract: Methods for determination of threshold values of signatures comprised in an assay are described. Each signature enables detection of a target. The methods determine a probability density function of negative samples and a corresponding false positive rate curve. A false positive criterion is established and a threshold for that signature is determined as a point at which the false positive rate curve intersects the false positive criterion. A method for quantitative analysis and interpretation of assay results together with a method for determination of a desired limit of detection of a signature in an assay are also described.

Abstract: System, including methods, apparatus, compositions, and kits, for making and using compound droplets of a multiple emulsion to supply an amplification reagent, such as a heat-stable DNA polymerase or DNA ligase, to an aqueous phase in which the compound droplets are disposed. The compound droplets may be induced to supply the amplification reagent by heating the multiple emulsion, to achieve hot-start amplification.

Abstract: Methods of characterizing a nucleic acid library by digital assay.

Abstract: Thermocycling system, including methods and apparatus, for performing a flow-based reaction on a sample in fluid. The system may include a plurality of segments defining at least two temperature regions, and also may include a plurality of heating elements configured to maintain each temperature region at a different desired temperature. At least one of the heating elements may be a thermoelectric cooler operatively disposed to transfer heat to and/or from a temperature region The system further may include a fluid channel extending along a helical path that passes through the temperature regions multiple times such that fluid flowing in the channel is heated and cooled cyclically.

Abstract: System, including methods and apparatus, for performing droplet-based assays that are controlled and/or calibrated using signals detected from droplets.

Abstract: Methods of processing an emulsion of aqueous droplets containing nucleic acid. The methods may include breakage of the emulsion with a destabilizing fluid including a halogen-substituted hydrocarbon.

Abstract: Methods to improve the performance of an array assay are described. A correlation between fluorescence intensity-related parameters and negative control values of the assay is determined. The parameters are then adjusted as a function of the correlation. As a result, sensitivity of the assay is improved without changes in its specificity.

Abstract: System, including methods and apparatus, for performing droplet-based assays that are controlled and/or calibrated using signals detected from droplets.

Abstract: System, including methods, apparatus, compositions, and kits, for assays with an emulsion including capsules. A method of performing an assay is provided. In the method, an aqueous phase may be provided. The aqueous phase may include a sample and an effective concentration of one or more skin-forming proteins. An emulsion may be formed. The emulsion may include droplets of the aqueous phase disposed in a nonaqueous continuous phase. The emulsion may be heated to create an interfacial skin between each droplet and the continuous phase, to transform the droplets into capsules. Assay data related to the sample may be collected from the capsules.

Abstract: Methods of characterizing a nucleic acid library by digital assay.

Abstract: System, including methods, apparatus, and compositions, for performing a multiplexed digital assay on a greater number of targets through combinatorial use of signals.

Abstract: A system for identifying all of the known and unknown pathogenic or non-pathogenic organisms in a sample. A droplet generator creates droplets from the sample. The droplets constitute sub-nanoliter volume reactors containing the organism sized particles. A lysis device performs lysis of the organisms to release the nucleic acids. An amplifier amplifies the nucleic acids. A fractionater releases the nucleic acids from the droplets. A parallel analyzer identifies all of the known and unknown pathogenic or non-pathogenic organisms in the sample.

Abstract: System, including methods and apparatus, for transporting droplets from a tip to an examination site for detection.