Abstract: This invention generally relates to methods and systems for dispensing a droplet, in particular a picolitre volume droplet containing one or more entities.

Abstract: The present invention provides a surfactant having a formula selected from the group consisting of: B—((X)x—(CH2)a-A)n(VI), (A-(CH2)a—(X)x—B—(X)x—(CH2)a-A)n (IV), (A-(CH2)a—(X)x—B)n (V), and (B)n—(X)x—(CH2)a-A (VII), wherein A is a perfluoropolyether; a is a positive integer; X is either a covalent bond or a linking group; x is a positive integer; B is a polyalkylene oxide unit; n is a positive integer greater than 1 and, in compounds comprising more than one A, B, X, a and x, each may be the same or different. The present invention also relates to methods of making such surfactants, uses of such surfactants and emulsions comprising such surfactants.

Abstract: A method for aligning sequences of droplets in streams of an emulsion comprising target droplets and tag droplets, a tag droplet comprising first and second tags. A target droplet is split into first and second target droplets and a tag droplet is split into first and second tag droplets. Each of the first and second tag droplets comprise the first and second tags. The first target droplet and first tag droplet are in a first stream of droplets, and the second target droplet and second tag droplet are in a second stream of droplets. The method detects the first tag droplets and first target droplets in the first stream and the second tag droplets and second target droplets in the second stream, determines a first sequence of droplets in the first stream and a second sequence of droplets in the second stream, and compares these to align the sequences.

Abstract: A method for aligning sequences of droplets in streams of an emulsion comprising target droplets and tag droplets, a tag droplet comprising first and second tags. A target droplet is split into first and second target droplets and a tag droplet is split into first and second tag droplets. Each of the first and second tag droplets comprise the first and second tags. The first target droplet and first tag droplet are in a first stream of droplets, and the second target droplet and second tag droplet are in a second stream of droplets. The method detects the first tag droplets and first target droplets in the first stream and the second tag droplets and second target droplets in the second stream, determines a first sequence of droplets in the first stream and a second sequence of droplets in the second stream, and compares these to align the sequences.

Abstract: We describe instruments, and cartridges therefor, for processing droplets of water in oil-in-water emulsions, the droplets containing biological entities such as cells. In one embodiment a method of such processing comprises: providing a plurality of the entities in a fluid: preparing a droplet from said fluid: determining whether said droplet contains one or more entities of said plurality of entities, or whether said droplet does not contain a said entity; sorting said droplet dependent on an outcome of said determination; and dispensing said sorted droplet into a reservoir, wherein said dispensing comprises: identifying said sorted droplet for dispensing; extracting said sorted droplet from a first fluidic flow path of said fluid by transferring said sorted droplet from said first fluidic flow path into a second fluidic flow path; and ejecting said sorted droplet from said second fluidic flow path into said reservoir by applying pressure to said second fluidic flow path.

Abstract: A method of processing droplets in a microfluidic system. The method may comprise capturing a time sequence of images of a droplet as it passes through a channel in a microfluidic system. The method may further comprise processing each image of the sequence of images using a convolutional neural network to count a number of cells or other entities visible in each image the droplet. The method may further comprise processing the count of the number of cells or other entities visible in each image of the droplet to determine an estimated number of cells or other entities in the droplet. The method/system may further comprise controlling a microfluidic process performed on the droplet responsive to the estimated number of cells or other entities in the droplet. Implementations of the method use the changing orientation and disposition of droplet contents in combination with machine learning to improve monoclonality assurance.

Abstract: Instruments and cartridges for processing droplets in emulsions containing biological entities such as cells. A method of such processing comprises providing a plurality of the entities in a fluid; preparing a droplet from the fluid; determining whether the droplet contains one or more entities of said plurality of entities, or whether said droplet does not contain a said entity; sorting said droplet dependent on an outcome of the determination; and dispensing the sorted droplet into a reservoir. The dispensing may comprise identifying and extracting the sorted droplet from a first fluidic flow path of said fluid by transferring the sorted droplet from into a second fluidic flow path and then ejecting the sorted droplet into a reservoir by applying pressure to the second fluidic flow path. The droplet contents may be tracked so that the contents of an individual droplet can be sorted, selectively dispensed, and retrieved.

Abstract: A method of performing a biological reaction in a microfluidic droplet, and in particular to a method of performing genome editing in a microfluidic droplet. The invention also relates to microfluidic systems and instrumentations or products for performing these reactions.

Abstract: This invention generally relates to methods and systems for dispensing a droplet, in particular a picolitre volume droplet containing one or more entities.

Abstract: A method for aligning sequences of droplets in streams of an emulsion comprising target droplets and tag droplets, a tag droplet comprising first and second tags. A target droplet is split into first and second target droplets and a tag droplet is split into first and second tag droplets. Each of the first and second tag droplets comprise the first and second tags. The first target droplet and first tag droplet are in a first stream of droplets, and the second target droplet and second tag droplet are in a second stream of droplets. The method detects the first tag droplets and first target droplets in the first stream and the second tag droplets and second target droplets in the second stream, determines a first sequence of droplets in the first stream and a second sequence of droplets in the second stream, and compares these to align the sequences.

Abstract: We describe a method comprising: providing a droplet comprising a plurality of constituents, splitting said droplet into a first droplet and a second droplet, wherein said first droplet comprises a first fraction of said plurality of constituents and said second droplet comprises a second fraction of said plurality of constituents, analysing said constituents of said first fraction of said plurality of constituents in said first droplet, and sorting said second droplet dependent on an outcome of said analysis.

Abstract: The present invention provides a surfactant having a formula selected from the group consisting of: B—((X)x—(CH2)a-Mn(VI), (A-(CH2)—(X)x—B—(X)x—(CH2)a-A)n (IV), (A-(CH2)a—(X)x—B)n (V), and (B)n—(X)x—(CH2)a—A (VII), wherein A is a perfluoropolyether; a is a positive integer; X is either a covalent bond or a linking group; x is a positive integer; B is a polyalkylene oxide unit; n is a positive integer greater than 1 and, in compounds comprising more than one A, B, X, a and x, each may be the same or different. The present invention also relates to methods of making such surfactants, uses of such surfactants and emulsions comprising such surfactants.

Abstract: A method of label-free cell or particle sorting in a microfluidic device includes providing a stream of aqueous droplets in oil in a channel of the microfluidic device, wherein at least some of the droplets include cells or particles and illuminating the stream from a first direction. Then, one detects scattered light from cells or particles within said aqueous droplets in a second direction. Next, one determines a number of the cells or particles in each droplet from the scattered light and sorts the aqueous droplets into differentiated streams. Notably, the refractive index of said oil is modified to closely match a refractive index of the aqueous droplets to reduce light scattered from boundaries of the droplets. A volume of the droplets can also controlled.

Abstract: Instruments and cartridges for processing droplets in emulsions containing biological entities such as cells. A method of such processing comprises providing a plurality of the entities in a fluid; preparing a droplet from the fluid; determining whether the droplet contains one or more entities of said plurality of entities, or whether said droplet does not contain a said entity; sorting said droplet dependent on an outcome of the determination; and dispensing the sorted droplet into a reservoir. The dispensing may comprise identifying and extracting the sorted droplet from a first fluidic flow path of said fluid by transferring the sorted droplet from into a second fluidic flow path and then ejecting the sorted droplet into a reservoir by applying pressure to the second fluidic flow path. The droplet contents may be tracked so that the contents of an individual droplet can be sorted, selectively dispensed, and retrieved.

Abstract: We describe a method comprising: providing a droplet comprising a plurality of constituents, splitting said droplet into a first droplet and a second droplet, wherein said first droplet comprises a first fraction of said plurality of constituents and said second droplet comprises a second fraction of said plurality of constituents, analysing said constituents of said first fraction of said plurality of constituents in said first droplet, and sorting said second droplet dependent on an outcome of said analysis.

Abstract: The present invention provides an emulsion for sorting droplets in a microfluidic device, the emulsion comprising: a discontinuous aqueous phase; and a continuous oil phase; wherein said aqueous phase comprises at least one analyte, said oil phase comprises a fluorous oil and a refractive index modifying compound comprising at least one aromatic ring, and the refractive index of said aqueous phase and the refractive index of said oil phase are substantially matched.

Abstract: We describe a method of label-free cell and/or particle sorting in a microfluidic device, the method comprising; providing a stream of aqueous droplets in oil in a channel of said microfluidic device, wherein at least some of said droplets include cells and/or particles illuminating said stream from a first direction; detecting, for example, scattered light from cells and/or particles within said aqueous droplets in a second direction; determining, for example, a number of said cells and/or particles in each droplet from the scattered light; and sorting said aqueous droplets into one of a plurality of differentiated streams responsive to said determined number. The method further comprises modifying a refractive index of said oil to more closely match a refractive index of said aqueous droplets to reduce light scattered from boundaries of said droplets. Optionally a volume of the droplets is also controlled. In embodiments the oil is fluorous oil.

Abstract: This invention relates to methods and systems for providing chemical and/or biological materials. Thus we describe a method of providing a chemical or biological material in a quantized form, the method comprising: preparing an emulsion comprising a plurality of microdroplets each holding a controlled amount of said material; storing said chemical or biological material in said microdroplets in said emulsion; transferring a portion of said emulsion microdroplets to a container for transportation; transporting said container to a user of said material, wherein said user receives said material in a quantized form in said microdroplets of said emulsion; and de-emulsifying said received microdroplets containing said material in quantized form to extract said material for subsequent use.

Abstract: The present invention provides an emulsion for sorting droplets in a microfluidic device, the emulsion comprising: a discontinuous aqueous phase; and a continuous oil phase; wherein said aqueous phase comprises at least one analyte, said oil phase comprises a fluorous oil and a refractive index modifying compound comprising at least one aromatic ring, and the refractive index of said aqueous phase and the refractive index of said oil phase are substantially matched.

Abstract: This invention relates to methods and systems for providing chemical and/or biological materials. Thus we describe a method of providing a chemical or biological material in a quantised form, the method comprising: preparing an emulsion comprising a plurality of microdroplets each holding a controlled amount of said material; storing said chemical or biological material in said microdroplets in said emulsion; transferring a portion of said emulsion microdroplets to a container for transportation; transporting said container to a user of said material, wherein said user receives said material in a quantised form in said microdroplets of said emulsion; and de-emulsifying said received microdroplets containing said material in quantised form to extract said material for subsequent use.