Process for manufacturing an array with microchannels

The invention relates to a process for manufacturing a microfluidic chip comprising a solid material obtained from a sol-gel solution, the process comprising successively: a) casting a sol-gel solution made with tetraethyl orthosilicate onto a mold presenting a relief pattern and having a different thickness over the whole of the mold; b) gelling the sol-gel solution; c) unmolding and drying the gel obtained in b), so as to obtain a solid glass; and d) bonding said solid glass to a support, so as to obtain the microfluidic chip.

SYSTEMS AND METHODS FOR PARTICULATE ENCAPSULATION IN MICRODROPLETS

The present invention generally relates to microfluidic droplets and, in particular, to multiple emulsion microfluidic droplets. Provided are methods and a device of ordering, sorting and/or focusing particles, the method comprising leading the particles through a microfluidic channel comprising a channel height (H) in the range of (1.8) D to (1.2) D and a channel width (W) in the range of (1.33) D to 1 D, wherein D is the particle diameter. 1. A method of ordering , sorting and/or focusing particles , the method comprising leading the particles through a microfluidic channel comprising a channel height (H) in the range of 1.8 D to 1.2 D , wherein D is the particle diameter.2. The method of claim 1 , wherein the microfluidic channel comprises a channel width (W) in the range of 1.33 D to 1 D.3. The method of claim 1 , wherein the microfluidic channel comprises a chamber for a particle reservoir claim 1 , wherein the chamber height requires 1.2 to 1.8 times the particle diameter and the chamber width is at least greater than twice the particle diameter.4. The method of claim 1 , wherein the particles are packed in the chamber before entering the microfluidic channel.5. The method of claim 3 , wherein the chamber comprises tapered lines leading to the microchannel.6. The method of claim 1 , wherein the microfluidic channel height is decreased at the exit.7. The method of claim 1 , wherein the inner wall of the microfluidic channel is hydrophobic.8. The method of claim 1 , wherein the particles are composed of a polymer material with an elastic modulus.9. The method of claim 1 , wherein the particles are hydrogel beads.10. The method of claim 1 , wherein the particles comprise capture molecules.11. The method of wherein the capture molecules may be selected from the group comprising claim 10 , an antigen claim 10 , an antibody or fragments thereof claim 10 , nucleic acids claim 10 , magnetic particles claim 10 , colloidal particles claim 10 , nanoparticles claim 10 , ...

SINGLE CELL ANALYSIS

The present invention concerns processes for barcoding nucleic acids from single cells and processes for genotyping single cells having a phenotype of interest. 1. A process for genotyping single cells having a phenotype of interest comprising: a) for a plurality of reservoirs, providing, for each reservoir, at least one barcode sequence and at least one dye, wherein the at least one barcode sequence is associated with the color and the concentration of said at least one dye, and taking an image of the array thereby mapping the color and the intensity of the at least one dye for each reservoir; and', 'b) for a plurality of reservoirs, providing, for each reservoir, at least one single cell and performing for each reservoir a phenotypic assay on said at least one single cell, and taking an image thereby mapping the phenotype of said at least one single cell for each reservoir;, 'providing a plurality of reservoirs, and'} 'c) for a plurality of reservoirs, linking, for each reservoir, the phenotype of a single cell obtained in step b) to the color and the intensity of the at least one dye obtained in step a); and', 'wherein step a) is performed before step b) or step b) is performed before step a); then'}for a plurality of reservoirs, reverse transcribing, for each reservoir, nucleic acids of said at least one single cell, to obtain single cell cDNA barcoded with said at least one barcode sequence which is associated with the color and concentration of the at least one dye; andlinking the genotype with the phenotype of said at least one single cell.2. The microfluidic process according to claim 1 , wherein said method further comprises at least one step selected from the group consisting of:at least one purging step,for a plurality of reservoirs, providing, for each reservoir, at least one oligonucleotide, wherein said at least one oligonucleotide comprises a primer sequence,for a plurality of reservoirs, attaching, for each reservoir, the at least one barcode ...

MICROFLUIDIC DEVICE

The invention relates to a microfluidic device including a chamber having a fluid inlet, a fluid outlet and a sealable port. In some embodiments, the fluid inlet and the fluid outlet may be positioned to direct fluid flowing from the fluid inlet to the fluid outlet through the chamber. Various embodiments may include a sealable port which may be aligned with the chamber to allow material to be placed directly into, or removed from, the chamber from the exterior of the device when the sealable port is open, and to inhibit and/or prevent fluid escaping through the sealable port when the port is sealed. 1. A microfluidic device comprising a chamber having a fluid inlet , a fluid outlet and a sealable port , wherein the fluid inlet and the fluid outlet are positioned to direct fluid flowing from the fluid inlet to the fluid outlet through the chamber , and wherein the sealable port is aligned with the chamber to allow material to be placed directly into , or removed from , the chamber from the exterior of the device when the sealable port is open , and to prevent fluid escaping through the sealable port when the port is sealed.2. The device of claim 1 , further comprising an interconnect system which comprises:a first component having a conduit to carry fluid to the fluid inlet or away from the fluid outlet, wherein the first component is formed of a deformable material, anda second component having a projecting portion, wherein a conduit passes through the projecting portion and the second component;wherein the conduit of the first component is aligned with the conduit of the second component, wherein the projecting portion of the second component deforms an area of the first component surrounding the conduit therein so as to create a seal around the contiguous conduits of the first and second components, thus preventing any fluid from escaping as it flows from one conduit to the other conduit, and wherein the second component is for connecting the conduit therein to ...

MICROFLUIDIC DEVICE

A multifunctional dual mode microfluidic device including a first operating configuration and a second operating configuration wherein in the first operating configuration, the device is configured to perform static cell seeding or produce cells. In the second operating configuration, the device is configured to perform perfusion via microfluidics of the device, and the device is configured to switch between the first operating configuration and the second operating configuration. The first operating configuration and the second operating configuration are selectively and independently accessible, wherein the first operating configuration or the second operating configuration does not alter any other configuration of the device. The device is fully operable in either one of the first operating configuration or the second operating configuration. 1. A microfluidic device comprising:a chamber having a fluid inlet, and a fluid outlet and a sealable port,wherein the fluid inlet and the fluid outlet are positioned to direct fluid flowing from the fluid inlet to the fluid outlet through the chamber, and wherein the sealable port is aligned with the chamber to allow material to be placed directly into, or removed from, the chamber from the exterior of the device when the sealable port is open, and to prevent fluid escaping through the sealable port when the sealable port is sealed, anda lid configured to seal the sealable port in a sealed position when the lid fills part of a volume of the chamber in a closed position, wherein the chamber has a smaller volume in the sealed position,wherein the lid is configured to unseal the sealable port in an unsealed position when the lid is removed from the part of a volume of the chamber in an open position, wherein the chamber has a larger volume in the open position,wherein the lid is in contact with the side walls of the chamber when the sealable port is closed,wherein the device is operable when the sealable port is open or closed ...

PARTICLE SORTING IN A MICROFLUIDIC SYSTEM

The invention relates to a method for ordering, sorting and/or focusing particles in a first microfluidic channel system, the method comprising the steps of i) providing for a first microfluidic channel comprising at least a first and a second inlet and a first outlet, ii) injecting a first fluid into the channel through said first inlet, iii) injecting a second fluid into the channel through said second inlet, wherein the viscosity of the first fluid is higher than the viscosity of the second fluid, such that the two fluids flow in a laminar fashion unmixed side by side, and one of the two fluids comprises the particles to be ordered, sorted and/or focused. The invention also relates to a microfluidic channel system for sorting different particles into one droplet. 1. A method for ordering , sorting and/or focusing particles in a first microfluidic channel system , the method comprising the steps ofi. providing for a first microfluidic channel comprising at least a first and a second inlet and a first outlet,ii. injecting a first fluid into the channel through said first inlet,iii. injecting a second fluid into the channel through said second inlet, wherein the viscosity of the first fluid is higher than the viscosity of the second fluid, such that the two fluids flow in a laminar fashion unmixed side by side, and one of the two fluids comprises the particles to be ordered, sorted and/or focused.2. A method according to claim 1 , wherein the particles are comprised preferably in the second fluid and the viscosity of the first fluid is selected such claim 1 , that the particles in the second fluid are confined by the first fluid to the space occupied by the second fluid.3. A method according to or claim 1 , wherein the height of the first microfluidic channel is selected from the group of between 2 μm and 60 μm claim 1 , 5 μm and 50 μm claim 1 , 10 μm and 45 μm claim 1 , 15 μm and 40 μm claim 1 , 25 μm and 35 μm.4. A method according to any of the to claim 1 , ...

Systems And Methods For Microfluidic Interfaces

The present invention is directed to a fluidic interface for delivering a fluid into a microfluidic device, the interface comprising a chamber comprising a first accepting/delivering fluid opening and a second accepting/delivering fluid opening, each opening being positioned opposite to the other, wherein the diameter of the first accepting/delivering fluid opening is smaller than the internal diameter of the chamber and a tubing system connecting the first accepting/delivering fluid opening to a pressure source capable of generating positive or negative pressure. The interface is characterized in that the second accepting/delivering fluid opening is designed to mechanically limit its insertion into a receptacle of a microfluidic device to ensure a predetermined gap (H) between the second accepting/delivering fluid opening and a lower side of the receptacle of a microfluidic device. 1. A fluidic interface for delivering a fluid into a microfluidic device , the interface comprising:a chamber comprising a first accepting/delivering fluid opening and a second accepting/delivering fluid opening, each opening being positioned opposite to the other, wherein the diameter of the first accepting/delivering fluid opening is smaller than the internal diameter of the chamber, anda tubing system connecting the first accepting/delivering fluid opening to a pressure source capable of generating positive or negative pressure,the interface being characterized in that the second accepting/delivering fluid opening is designed to mechanically limit its insertion into a receptacle of a microfluidic device to ensure a predetermined gap (H) between the second accepting/delivering fluid opening and a lower side of the receptacle of a microfluidic device.2. The fluidic interface according to the claim 1 , wherein said chamber and tubing system comprise a first type of fluid.3. The fluidic interface according to the claim 2 , wherein said first type of fluid is a hydraulic fluid.4. The ...

METHOD FOR TRANSCRIPTOME ANALYSIS OF SINGLE CELLS

The present invention concerns a method for capturing and barcoding nucleic acid from single cells, a plurality of microfluidic droplets and a method for preparing said plurality of microfluidic droplets. 1. A method for capturing and barcoding single cell nucleic acid comprising:a) providing a plurality of cells contained within a plurality of compartments, at least some of the compartments comprising a single cell, a reverse transcriptase and at least one type of an oligonucleotide, wherein the at least one type of oligonucleotide comprises a barcode sequence and a primer sequence, wherein each different primer sequence defines a different oligonucleotide type, and wherein the compartments of the plurality of the compartments contain one or more barcode sequences distinguishable from barcode sequences contained in other compartments of the plurality of compartments;b) lysing at least some of the cells within the compartments to release nucleic acids from the cells;c) hybridizing at least some of the released nucleic acids to said oligonucleotide in at least some of the compartments;d) reverse transcribing the released nucleic acids hybridized to said oligonucleotide using the primer sequence in at least some of the compartments;wherein the plurality of compartments has a volume of less than 3 nl_.2. The method according to claim 1 , wherein the concentration of each type of oligonucleotide in the compartments is at least 100 nM.3. The method according to claim 1 , wherein step a) further contains providing a plurality of particles contained within said plurality of compartments claim 1 , and wherein at least some of the compartments further comprise a particle.4. The method according to claim 3 , wherein the at least one type of an oligonucleotide is bonded to said particle.5. The method according to claim 1 , wherein the plurality of compartments has a volume equal to or less than 1 nl_.6. The method according to claim 1 , which further comprises recovering ...

METHOD FOR ANALYZING AND SELECTING A SPECIFIC DROPLET AMONG A PLURALITY OF DROPLETS AND ASSOCIATED APPARATUS

The present invention relates to a method for analyzing and selecting a specific droplet among a plurality of droplets (), comprising the following steps: —providing a plurality of droplets (), —for a droplet () among the plurality of droplets, measuring at least two optical signals, each optical signal being representative of a light intensity spatial distribution in the droplet for an associated wavelength channel, —calculating a plurality of parameters from the optical signals, —determining a sorting class for a droplet according to calculated parameters, —sorting said droplet according to its sorting class, wherein the plurality of parameters comprises the coordinates of a maximum for each optical signal and a co-localization parameter and the at least two calculated parameters used for the determining step comprises the co-localization parameter. 2. A method according to claim 1 , wherein a co-localization parameter (Δ) is calculated by comparing the position corresponding to the maximum intensity of a first optical signal among the at least two optical signals to the position corresponding to the maximum intensity of a second optical signal among the at least two optical signals.3. A method according to any of the and claim 1 , wherein the co-localization parameter (Δ) is selected by having a confidence interval ranging from 90% to 100%.4. A method according to any of the to claim 1 , wherein the plurality of parameters comprises at least one of the following parameters:{'b': '4', 'a droplet () width,'}an integration of an optical signal,a ratio between a maximum value of an optical signal and an integration value of said optical signal,the coordinates of a local maximum for an optical signal,the number of local maxima in a droplet for an optical signal,the calculation of the derivative of an optical signal andthe calculation of the second derivative for an optical signal.5. A method according to any of the preceding claims claim 1 , wherein a first optical ...

Particle sorting in a microfluidic system

The invention relates to a method for ordering, sorting and/or focusing particles in a first microfluidic channel system, the method comprising the steps of i) providing for a first microfluidic channel comprising at least a first and a second inlet and a first outlet, ii) injecting a first fluid into the channel through said first inlet, iii) injecting a second fluid into the channel through said second inlet, wherein the viscosity of the first fluid is higher than the viscosity of the second fluid, such that the two fluids flow in a laminar fashion unmixed side by side, and one of the two fluids comprises the particles to be ordered, sorted and/or focused. The invention also relates to a microfluidic channel system comprising i) a first microfluidic channel comprising at least a first and a second inlet and a first outlet, ii) a first fluid, iii) a second fluid, wherein the viscosity of the first fluid is higher than the viscosity of the second fluid, such that the two fluids can flow in a laminar fashion unmixed side by side, and one of the two fluids comprises particles to be ordered, sorted and/or focused.

Single cell analysis

The present invention concerns processes for barcoding nucleic acids from single cells and processes for genotyping single cells having a phenotype of interest.

Systems and methods for microfluidic interfaces

The present invention is directed to a fluidic interface for delivering a fluid into a microfluidic device, the interface comprising a chamber comprising a first accepting/delivering fluid opening and a second accepting/delivering fluid opening, each opening being positioned opposite to the other, wherein the diameter of the first accepting/delivering fluid opening is smaller than the internal diameter of the chamber and a tubing system connecting the first accepting/delivering fluid opening to a pressure source capable of generating positive or negative pressure. The interface is characterized in that the second accepting/delivering fluid opening is designed to mechanically limit its insertion into a receptacle of a microfluidic device to ensure a predetermined gap (H) between the second accepting/delivering fluid opening and a lower side of the receptacle of a microfluidic device.

Method for transcriptome analysis of single cells

The present invention concerns a method for capturing and barcoding nucleic acid from single cells, a plurality of microfluidic droplets and a method for preparing said plurality of microfluidic droplets.

Single cell analysis

The present invention concerns processes for barcoding nucleic acids from single cells and processes for genotyping single cells having a phenotype of interest.

Microfluidic methods and systems

The invention relates to a microfluidic system comprising:a) a solid support comprising at least a first group of oligonucleotides,i. wherein each oligonucleotide in said group comprises a nucleic acid sequence of a first type, of a second type and/or a further type,ii. wherein said nucleic acid sequence of a first type is a barcode sequence,iii. and oligonucleotides comprising the same barcode sequence are grouped together in a group of oligonucleotides on said solid support,iv. wherein the first and further oligonucleotide groups are spatially separated on said solid support,b) wherein said one or more groups of oligonucleotide groups on said solid support are within separate reservoirs of the microfluidics system,c) wherein the one or more reservoirs are accessible to fluids, cells, chemicals and/or microdroplet by means of channels, andd) wherein each reservoir comprises comprising a group of oligonucleotides on said solid support is also trap for a microfluidic droplet.

Method for analyzing and selecting a specific droplet among a plurality of droplets and associated apparatus

The present invention relates to a method for analyzing and selecting a specific droplet among a plurality of droplets (4), comprising the following steps: - providing a plurality of droplets (4), - for a droplet (4) among the plurality of droplets, measuring at least two optical signals, each optical signal being representative of a light intensity spatial distribution in the droplet for an associated wavelength channel, - calculating a plurality of parameters from the optical signals, - determining a sorting class for a droplet according to calculated parameters, - sorting said droplet according to its sorting class, wherein the plurality of parameters comprises the coordinates of a maximum for each optical signal and a co-localization parameter and the at least two calculated parameters used for the determining step comprises the co-localization parameter.

Method for transcriptome analysis of single cells

The present invention concerns a method for capturing and barcoding nucleic acid from single cells, a plurality of microfluidic droplets and a method for preparing said plurality of microfluidic droplets.

Single cell analysis

Systems and methods for microfluidic interfaces

The present invention is directed to a fluidic interface for delivering a fluid into a microfluidic device, the interface comprising a chamber comprising a first accepting/delivering fluid opening and a second accepting/delivering fluid opening, each opening being positioned opposite to the other, wherein the diameter of the first accepting/delivering fluid opening is smaller than the internal diameter of the chamber and a tubing system connecting the first accepting/delivering fluid opening to a pressure source capable of generating positive or negative pressure. The interface is characterized in that the second accepting/delivering fluid opening is designed to mechanically limit its insertion into a receptacle of a microfluidic device to ensure a predetermined gap (H) between the second accepting/delivering fluid opening and a lower side of the receptacle of a microfluidic device.

Particle sorting in a microfluidic system

The invention relates to a method for ordering, sorting and/or focusing particles in a first microfluidic channel system, the method comprising the steps of i) providing for a first microfluidic channel comprising at least a first and a second inlet and a first outlet, ii) injecting a first fluid into the channel through said first inlet, iii) injecting a second fluid into the channel through said second inlet, wherein the viscosity of the first fluid is higher than the viscosity of the second fluid, such that the two fluids flow in a laminar fashion unmixed side by side, and one of the two fluids comprises the particles to be ordered, sorted and/or focused. The invention also relates to a microfluidic channel system comprising i) a first microfluidic channel comprising at least a first and a second inlet and a first outlet, ii) a first fluid, iii) a second fluid, wherein the viscosity of the first fluid is higher than the viscosity of the second fluid, such that the two fluids can flow in a laminar fashion unmixed side by side, and one of the two fluids comprises particles to be ordered, sorted and/or focused.

Microfluidic methods and systems

The invention relates to a microfluidic system comprising:a) a solid support comprising at least a first group of oligonucleotides,i. wherein each oligonucleotide in said group comprises a nucleic acid sequence of a first type, of a second type and/or a further type,ii. wherein said nucleic acid sequence of a first type is a barcode sequence,iii. and oligonucleotides comprising the same barcode sequence are grouped together in a group of oligonucleotides on said solid support,iv. wherein the first and further oligonucleotide groups are spatially separated on said solid support,b) wherein said one or more groups of oligonucleotide groups on said solid support are within separate reservoirs of the microfluidics system,c) wherein the one or more reservoirs are accessible to fluids, cells, chemicals and/or microdroplet by means of channels, andd) wherein each reservoir comprises comprising a group of oligonucleotides on said solid support is also trap for a microfluidic droplet.

Microfluidic methods and systems

The invention relates to a microfluidic system comprising:a) a solid support comprising at least a first group of oligonucleotides,i. wherein each oligonucleotide in said group comprises a nucleic acid sequence of a first type, of a second type and/or a further type,ii. wherein said nucleic acid sequence of a first type is a barcode sequence,iii. and oligonucleotides comprising the same barcode sequence are grouped together in a group of oligonucleotides on said solid support,iv. wherein the first and further oligonucleotide groups are spatially separated on said solid support,b) wherein said one or more groups of oligonucleotide groups on said solid support are within separate reservoirs of the microfluidics system,c) wherein the one or more reservoirs are accessible to fluids, cells, chemicals and/or microdroplet by means of channels, andd) wherein each reservoir comprises comprising a group of oligonucleotides on said solid support is also trap for a microfluidic droplet.

Process for manufacturing an array with microchannels

The invention relates to a process for manufacturing a microfluidic chip comprising a solid material obtained from a sol-gel solution, the process comprising successively: a) casting a sol-gel solution made with tetraethyl orthosilicate onto a mold presenting a relief pattern and having a different thickness over the whole of the mold; b) gelling the sol-gel solution; c) unmolding and drying the gel obtained in b), so as to obtain a solid glass; and d) bonding said solid glass to a support, so as to obtain the microfluidic chip.

Process for manufacturing an array with microchannels

The invention relates to a process for manufacturing a microfluidic chip comprising a solid material obtained from a sol-gel solution, the process comprising successively: a) casting a sol-gel solution made with tetraethyl orthosilicate onto a mold presenting a relief pattern and having a different thickness over the whole of the mold; b) gelling the sol-gel solution; c) unmolding and drying the gel obtained in b), so as to obtain a solid glass; and d) bonding said solid glass to a support, so as to obtain the microfluidic chip.

Method for analyzing and selecting a specific droplet among a plurality of droplets and associated apparatus

The present invention relates to a method for analyzing and selecting a specific droplet among a plurality of droplets (4), comprising the following steps: - providing a plurality of droplets (4), - for a droplet (4) among the plurality of droplets, measuring at least two optical signals, each optical signal being representative of a light intensity spatial distribution in the droplet for an associated wavelength channel, - calculating a plurality of parameters from the optical signals, - determining a sorting class for a droplet according to calculated parameters, - sorting said droplet according to its sorting class, wherein the plurality of parameters comprises the coordinates of a maximum for each optical signal and a co-localization parameter and the at least two calculated parameters used for the determining step comprises the co-localization parameter.

Microfluidic methods and systems

The invention relates to a microfluidic system comprising: a) a solid support comprising at least a first group of oligonucleotides, i. wherein each oligonucleotide in said group comprises a nucleic acid sequence of a first type, of a second type and/or a further type, ii. wherein said nucleic acid sequence of a first type is a barcode sequence, iii. and oligonucleotides comprising the same barcode sequence are grouped together in a group of oligonucleotides on said solid support, iv. wherein the first and further oligonucleotide groups are spatially separated on said solid support, b) wherein said one or more groups of oligonucleotide groups on said solid support are within separate reservoirs of the microfluidics system, c) wherein the one or more reservoirs are accessible to fluids, cells, chemicals and/or microdroplet by means of channels, and d) wherein each reservoir comprises comprising a group of oligonucleotides on said solid support is also trap for a microfluidic droplet.

Single cell analysis

The present invention concerns processes for barcoding nucleic acids from single cells and processes for genotyping single cells having a phenotype of interest.

Single cell analysis

The present invention concerns processes for barcoding nucleic acids from single cells and processes for genotyping single cells having a phenotype of interest.