Stokes Bio Limited

Forfas

Forf&agrave;s

Hooke Bio Limited

Mark Davies

Tara Dalton

Kieran Curran

Damian Michael Curtin

Brian T Chawke

Julie Garvey

Michael Sayers

Conor McCarthy

David McGuire

Brian Barrett

Noel Sirr

Ryan J Talbot

Mauro Aguanno

David Kinahan

Jeffrey Punch

Gerard O'Regan

Damien King

Philip Samways

Martin J Leahy

Catherine Byrne

Stephen Clothier

Mark Korenke

Xiaona Hou

John Daly

David Mcguire

Michael Galvin

A system for processing a biological sample can include a droplet generation assembly comprising a plurality of first reservoirs configured to contain an aqueous sample and a plurality of second reservoirs configured to contain a carrier fluid immiscible with the aqueous sample. The plurality of first reservoirs and the plurality of second reservoirs can be arranged to be in respective flow communication in pairs of reservoirs comprising a first reservoir of the plurality of first reservoirs and a second reservoir of the plurality of second reservoirs constituting a plurality of pairs of reservoirs. The droplet generation assembly can further include a flow control system configured to control a pressure in the plurality of pairs of reservoirs so as to generate a flow of a series of volumes of the aqueous sample separated by the carrier fluid. The system can further include a thermocycling system.

Liquid bridge and system

A system for analyzing a biological sample may include at least one sample acquisition stage comprising a sample acquisition device for acquiring the biological sample from a sample source; a droplet generator device for forming a droplet wrapped in an immiscible carrier fluid, wherein the wrapped droplet comprises at least the biological sample and a reagent, the droplet generator configured to receive the biological sample transferred from the sample acquisition device; a collection vessel for collecting the wrapped sample droplet from the droplet generator, the vessel configured to contain a carrier fluid for receiving and protecting the sample droplet; and an analysis system for analyzing the wrapped sample droplet and detecting products of a polymerase chain reaction.

Methods of releasing and analyzing cellular components

A microfluidic analysis system () performs polymerase chain reaction (PCR) analysis on a bio sample. In a centrifuge () the sample is separated into DNA and RNA constituents. The vortex is created by opposing flow of a silicon oil primary carrier fluid effecting circulation by viscous drag. The bio sample exits the centrifuge enveloped in the primary carrier fluid. This is pumped by a flow controller () to a thermal stage (). The thermal stage () has a number of microfluidic devices () each having thermal zones () in which the bio sample is heated or cooled by heat conduction to/from a thermal carrier fluid and the primary carrier fluid. Thus, the carrier fluids envelope the sample, control its flowrate, and control its temperature without need for moving parts at the micro scale.

Microfluidic analysis system

A system for processing a biological sample includes a substrate comprising a plurality of wells and a plurality of flow channels. The system further includes a flow control system comprising a manifold having a plurality of ports configured to fluidically couple to the plurality of wells, and one or more containment structures configured to contain carrier fluid and fluidically couple to the ports. The flow control system further includes a cradle configured to removably receive the substrate. The flow control system is configured to transmit pressure differential, via the manifold, to the plurality of wells so as to cause a plurality of sample volumes held by at least some wells of the plurality of wells to flow through respective flow channels and cause the carrier fluid to flow through the flow channels and form a plurality of droplets of the plurality of sample volumes separated by the carrier fluid.

System for processing biological sample

A biological sample analysis system including a sample preparation system forming droplets of segmented sample separated by a carrier fluid immiscible with the sample. The droplets include reaction mixtures for amplification of at least one target nucleic acid. A thermal cycling device having a sample block having a plurality of controlled thermal zones, and a containment structure in thermal communication with the plurality of controlled thermal zones. The containment structure receives and contains the droplets of segmented sample separated by the immiscible carrier fluid from the sample preparation system. A controller for controlling a temperature in each thermal zone of the sample block. A detection system detects electromagnetic radiation emitted from each of the droplets individually from the queue of droplets as they flow past the detection system. A positioning system to facilitate moving a queue of the droplets in the thermal cycling device relative to the detection system.

A bridge () comprises a first inlet port () at the end of a capillary, a narrower second inlet port () which is an end of a capillary, an outlet port () which is an end of a capillary, and a chamber () for silicone oil. The oil is density-matched with the reactor droplets such that a neutrally buoyant environment is created within the chamber (). The oil within the chamber is continuously replenished by the oil separating the reactor droplets. This causes the droplets to assume a stable capillary-suspended spherical form upon entering the chamber (). The spherical shape grows until large enough to span the gap between the ports, forming an axisymmetric liquid bridge. The introduction of a second droplet from the second inlet port () causes the formation of an unstable funicular bridge that quickly ruptures from the, finer, second inlet port (), and the droplets combine at the liquid bridge (). In another embodiment, a droplet () segments into smaller droplets which bridge the gap between the inlet and outlet ports.

Methods of conducting a nucleic acid reaction, including methods for performing digital PCR using a 'droplet-in-oil' technology, may include at least partially segmenting a starting sampled into a set of sample droplets each containing on average about one or fewer copies of a target nucleic acid. The droplets are passed in a continuous flow of immiscible carrier fluid through a channel that passes through a thermal cycler, whereby the target is amplified. In one implementation, the droplets are about 350 nl each and the number of positively amplified droplets is counted at the near-saturation point.

Methods and devices for digital PCR

Provided herein is a biological detection system and method of use wherein the biological detection system comprises at least one mixer or liquid bridge for combining at least two liquid droplets and an error correction system for detecting whether or not proper mixing or combining of the two component droplets have occurred.

Biological detection system and method of use

The invention provides methods of conducting a nucleic acid reaction, including methods for performing digital PCR using a 'droplet-in-oil' technology. In the methods, the starting sampled is segmented at least partially into a set of sample droplets each containing on average about one or fewer copies of a target nucleic acid. The droplets are passed in a continuous flow of immiscible carrier fluid through a channel that passes through a thermal cycler, whereby the target is amplified. In one implementation, the droplets are about 350 nl each and the number of positively amplified droplets is counted at the near-saturation point.

Provided herein is a method for analyzing the affect of at least one agent on a cell or cellular component using an analysis system. The method includes wrapping a droplet in an immiscible carrier fluid using a droplet generator wherein the wrapped droplet includes at least one cell or cellular component and at least one agent. The wrapped droplet then flows through the analysis system using a siphoning effect. The droplet is then analyzed to determine effect of the agent on the cell or cellular component.

Methods for culturing and analyzing cells

The present invention generally relates to methods of constructing liquid bridges and methods of forming predetermined combinations of samples using liquid bridges.

Forming sample combinations using liquid bridge systems

The present invention generally relates to devices, systems, and methods for acquiring and/or dispensing a sample without introducing a gas into a microfluidic system, such as a liquid bridge system. An exemplary embodiment provides a sampling device including an outer sheath; a plurality of tubes within the sheath, in which at least one of the tubes acquires a sample, and at least one of the tubes expels a fluid that is immiscible with the sample, in which the at least one tube that acquires the sample is extendable beyond a distal end of the sheath and retractable to within the sheath; and a valve connected to a distal portion of the sheath, in which the valve opens when the tube extends beyond the distal end and closes when the tube retracts to within the sheath.

Sampling device

A bridge comprises a first inlet port, a second inlet port, an outlet port, and a chamber for silicone oil. The oil is density-matched with the reactor droplets such that a neutrally buoyant environment is created within the chamber. The oil within the chamber is continuously replenished by the oil separating the reactor droplets. This causes the droplets to assume a stable capillary-suspended spherical form upon entering the chamber. The spherical shape grows until large enough to span the gap between the ports, forming an axisymmetric liquid bridge. The introduction of a second droplet from the second inlet port causes the formation of an unstable funicular bridge that quickly ruptures from the, finer, second inlet port, and the droplets combine at the liquid bridge. In another embodiment, a droplet segments into smaller droplets which bridge the gap between the inlet and outlet ports.

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Limerick, Ireland

Mark Davies