Improvements in devices and methods for dispensing droplets, or related improvements.

Abstract

The present invention provides a device, method, and apparatus for dispensing one or more microdroplets. [Solution] The device comprises a microfluidic chip having an oEWOD structure configured to generate an optically mediated electrowetting (oEWOD) force. The microfluidic chip includes a first region and a second region. The first and second regions are separated by a constriction. The first region is adapted to receive and manipulate one or more microdroplets dispersed in a carrier fluid at a first flow rate. The second region is configured to receive the microdroplets from the first region via the constriction and move the microdroplets to the outlet port of the microfluidic chip at a second flow rate. The second region is configured to receive the microdroplets from the first region via the constriction by the application of an optically mediated electrowetting (oEWOD) force.

Description

[Technical Field]

[0001] The present invention relates to a device and method for dispensing microdroplets, particularly one or more microdroplets The present invention relates to a device comprising a microfluidic tip for dispensing a microfluidic. The present invention also relates to one or more microfluidic tips. This concerns a method for dispensing small droplets. [Background technology]

[0002] Devices for manipulating droplets or magnetic beads are known in the art. One technique for the operation is, for example, to move a droplet in the presence of an immiscible carrier fluid. The microfluidic space defined by two opposing walls of a ridge or microfluidic tube This includes moving through the wall. Microelectrodes embedded in one or both walls induce Covered by an electric layer, each dielectric layer is rapidly swept at intervals to correct the electric field characteristics of the layer. It connects to an A / C bias circuit that can be switched on and off. The vicinity of a microelectrode may be used to steer a droplet along one or more predetermined paths. This generates localized, directional capillary force in the following context. Hereinafter, in relation to the present invention, "real" refers to the real )Such devices that use what are called electrowetting electrodes are abbreviated The EWOD (Electrowetting on Dielectric) device is known in the art. A variation of this approach in which electrowetting forces are optically mediated is the technology. In the field, this is known as optoelectrowetting, and Below, we will refer to them as oEWOD, using the corresponding acronyms.

[0003] Microfluidic devices using oEWOD are defined by the first and second walls. It may include a small fluid cavity. The first wall is a composite design, consisting of a substrate, a photoconductive layer and an insulating layer. It is composed of a dielectric layer. Between the photoconductive layer and the insulating layer, there is an electrical insulation between them, and light An array of conductive cells bonded to the active layer is arranged. Its function corresponds to that of the insulating layer. The objective is to generate the positions of the electrowetting electrodes. At these positions, the electro The surface tension characteristics of the droplets can be changed using a trowetting field. These conductive cells can be temporarily switched on by light hitting the photoconductive layer. Yes. This approach has the advantage of making switching easier and faster. Its usefulness is still somewhat limited by the arrangement of the electrodes. Furthermore, transferring the droplet There are limitations regarding the movable speed and the range over which the actual droplet path can be varied.

[0004] Droplet manipulation using EWOD or oEWOD in microfluidic chips as described above During and / or after, many of the expected workflows on microfluidic systems involve cells, Materials such as beads or genetic material are extracted from the microfluidic chip and placed in a 384-well plate. It is necessary to collect the microfluidic fluid in a conventional liquid processing container such as a tub or microtube. The droplets dispensed from the tip can be further assayed. These assays include These generally include PCR amplification, DNA sequencing, RNA sequencing, and cell expansion. In particular, droplet recovery is often necessary for gene assays because microstreams When performed in a body chip, such assays can be performed on any cells held on the chip. This is because extreme temperature cycles that would kill it are typically involved.

[0005] The recovery of sub-nanoliter droplets from microfluidic systems has been an engineering challenge in microfluidics for many years. Generally, the required volume displacement mechanically constrains the actuator used to displace the fluid. Therefore, it has been difficult or impossible to recover droplets one by one through conventional mechanical operations. Existing systems, which are known for continuous-flow fluid engineering, include drop-on-demand microactuators and precisely designed dispensing nozzles. Basically, each requires a nanoliter fluid displacement step. Existing systems that are known for continuous-flow fluid engineering include drop-on-demand microactuators and precisely designed dispensing nozzles. Basically, each requires a nanoliter fluid displacement step.

[0006] An alternative approach to single-droplet recovery is the use of barcoding chemicals such as DNA barcodes. In this class of schemes, droplets are introduced into the droplet fluid system and loaded with unique DNA barcodes before being assayed. DNA sequencing often requires costly and complex instrumentation. Next, droplets of interest in the on-chip assay are recovered in a pooled format. The barcodes are read to recover the identity of the input cells. Such a scheme avoids the need for droplet-by-droplet recovery. However, they impose constraints on the nature of the on-chip assay and add costly and complex preparation and analysis steps.

[0007] Therefore, there is a need to provide a droplet recovery system for users that can be easily implemented in combination with a microfluidic chip. In addition, materials are recovered from the chip to To perform assays related to this, we move microdroplets from a microfluidic chip, cost-effectively. There is also a need to provide highly efficient dispensing systems and methods. It has the flexibility to collect Toll droplets one by one, while also having pooled for when necessary. There is also a need for a system that can dispense droplets onto a mat. [Overview of the Initiative] [Problems that the invention aims to solve]

[0008] This invention arose in response to the above-mentioned background. [Means for solving the problem]

[0009] According to one aspect of the present invention, optically mediated electrowetting (oEWOD) ) comprising a microfluidic chip having an oEWOD structure configured to generate force, A device for dispensing one or more microdroplets is provided. The microfluidic chip is in the first region and a second region, the first and second regions being separated by a constriction, The first region receives one or more microdroplets dispersed in the carrier fluid at a first flow rate. Adapted to be taken and operated, The second region receives minute droplets from the first region through the constricted portion, and the second region flows the minute droplets through the second flow. It is configured to move a certain amount of fluid to the exit port of the microfluidic chip. The second region is the application of optically mediated electrowetting (oEWOD) forces. Therefore, it is configured to receive the minute droplet from the first region through the constricted portion. The second flow rate in the second region is higher than the first flow rate in the first region.

[0010] In some embodiments, one or more microdroplets comprising a microfluidic tip are separated. A device for note-taking may be provided. The microfluidic chip includes a first region and a second region. The first and second regions are separated by a constricting means. The first region is a low-carrier fluid in which one or more microdroplets dispersed in a carrier fluid are placed. It is adapted to receive and operate by flow rate. The second region receives the microdroplets from the first region via the constriction means, and the microdroplets Configured to move the carrier fluid to the outlet port of the microfluidic chip at a higher carrier fluid flow rate. In the device, The second region is the application of optically mediated electrowetting (oEWOD) forces. Therefore, it is configured to receive the minute droplets from the first region via the constricting means. It is characterized by.

[0011] The devices and methods disclosed in the present invention, as described in the present invention, are for microdroplets and In some cases, the recovery of sub-nanol liter droplets from microfluidic systems is simplified. This is advantageous for enabling cost-effective systems. The recovery system allows users to efficiently remove droplets from microfluidic devices and achieve their desired results. The droplets can be collected onto a receptacle such as a multiwell plate, or dispensed. This is possible. This allows users to perform droplet microscopy, which is not easily done on microfluidic chips. Further assays can be performed to determine the cell content or bead content. The assay involves PCR amplification, DNA sequencing, RNA sequencing, and / or cell expansion. This may include, but is not limited to, individual droplets from microfluidic devices. These can be selected for recovery and then deposited onto a multi-well plate.

[0012] Furthermore, after dispensing individual droplets using the disclosed device and method, a preliminary screen is used. Leaning can be performed to select only the desired microdroplets. This allows for subsequent selection. Analysis of irrelevant droplets is prevented, and only the relevant subsections of the on-chip droplet are selected. This will become possible.

[0013] In addition, a subgroup of droplets contained within the microfluidic device can be selected for dispensing. This is possible. On the other hand, the remaining droplets do not necessarily affect those environmental conditions. It is held within the chip.

[0014] In some embodiments, the microdroplets may be dispensed individually from the apparatus. In one embodiment, multiple microdroplets may be dispensed simultaneously from the device. In this embodiment, the microdroplets may be grouped or pooled by activity. Multiple selected microdroplets may be dispensed from the device as needed. In this embodiment, the activity of the microdroplets may be addressed, for example, by fluorescence intensity.

[0015] In some embodiments, the carrier fluid in the first region is low flow or zero flow. It is a quantity. The first region can be used to hold or store microdroplets. Droplet manipulation in this context involves, for example, sorting, combining, dividing, or arranging droplets in an array. This may include, but is not limited to, oEWOD operations for the purpose of performing the following actions.

[0016] In some cases, microdroplets can be manipulated in the first region of the tip. In some embodiments, the low flow rate may be in the range of 0 to 20 μL / min. In one embodiment, the first flow rate may be in the range of 0 to 20 μL / min.

[0017] In some embodiments, the carrier fluid in the second region has a high flow rate. By supplying a high flow rate to two regions, the droplets are directed toward the outlet port of the microfluidic device. It can move in that way. For example, high flow rates are in the range of 10 to 100 μL / min. For example, the second flow rate is in the range of 10 to 100 μL / min.

[0018] In some embodiments, the rate is low / zero while receiving droplets. The rate can change at a higher rate while droplets or multiple microdroplets are being discharged. The flow rate in two regions can be dynamically controlled. In a further embodiment, droplets Unless otherwise noted, the flow rate in the second region may be 0-20 μL / min. In the embodiment, the flow rate in the second region is 10 to 100 μL / min during the dispensing procedure. That's fine.

[0019] In some embodiments, the second region receives multiple microdroplets from the first region. The flow rate in the region may be 0.02 to 2.00 μL / min. Alternatively, in the second region The flow rates are 0.02, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, and 0.6. It may exceed 0.7, 0.8, 0.9, 1, 1.5, or 2 μL / min. In the application configuration, the flow rates in the second region were 2, 1.5, 1, 0.9, 0.8, 0.7. Less than 0.6, 0.5, 0.4, 0.3, 0.2, 0.1, or 0.05 μL / min This is also acceptable. When the flow rate in the second region exceeds 0 μL / min, it is possible to prevent the micro-droplets from blocking the constriction portion. Subsequently, when the second region receives a plurality of micro-droplets at once, the flow rate can be increased to efficiently dispense the micro-droplets from the fluid device. In some embodiments, before the flow rate in the second region is increased, the second region may receive 1 to 10,000 micro-droplets. In some examples, the second region may receive more than 1, 50, 100, 200, 500, 700, 1000, 1250, 1500, 1750, 2000, 2500, 3000, 3500, 4500, 5000, 5500, 6000, 6500, 7500, 8000, 8500, 9000, 9500 or 10,000 micro-droplets. In some embodiments, the flow rate in the second region can be increased to exceed 2, 5, 10, 15 or 20 μL / min. This can be prevented. Subsequently, when the second region receives a plurality of micro-droplets at once, the flow rate can be increased to efficiently dispense the micro-droplets from the fluid device. This can be efficiently dispensed from the fluid device by increasing the flow rate. In some embodiments, before the flow rate in the second region is increased, the second region may receive 1 to 10,000 micro-droplets. In some examples, the second region may receive more than 1, 50, 100, 200, 500, 700, 1000, 1250, 1500, 1750, 2000, 2500, 3000, 3500, 4500, 5000, 5500, 6000, 6500, 7500, 8000, 8500, 9000, 9500 or 10,000 micro-droplets. This may exceed micro-droplets at 100, 200, etc. This may exceed micro-droplets at 2000, 2500, etc. This may exceed micro-droplets at 6500, 7500, etc. In some embodiments, the flow rate in the second region can be increased to exceed 2, 5, 10, 15 or 20 μL / min. This can be increased to exceed...

[0020] In some embodiments, the cross-sectional area of the first region may be 1×10 9 , 10 , 8 , 2 , 8 , , 10 , 9 , , 9 , , 8 , 8 , 9 , 2 , 10 , 10 , , 9 ~1.3×10 10 [[ID=X]]μm 2 In some embodiments, the area of the first region may be 1×10 <XXXXX>、2.5 ×10 <XXXXX>、5×10 8 、7.5×10 <00000XX>、1×10 / / Should be 9 <00XXXXX>、2.5×10 9 、5×10 9 、7.5× 10 9 、1×10 10 または1.25×10 10 μm 2 This may exceed... In some embodiments, the area of the first region may be 1.3×10 10 、1×10 10 、7.5×10 9 It seems there are some incorrect or incomplete tags in the original text which might affect the accuracy of the translation. Please check and correct the original text for a more precise translation. The above translation is based on the best understanding of the provided content., 5×1 0 9 , 2.5×10 9 , 1 x 10 9 , 7.5×10 8 , 5×10 8 or 2.5 × 10 8 μm 2 It is acceptable to be less than [a certain value].

[0021] In some embodiments, the area of ​​the first region may be larger than the area of ​​the second region. i. It is advantageous that the first region has a large cross-sectional area, allowing for efficient manipulation of a large number of droplets. This makes high-throughput devices easier to implement. The first area may accommodate multiple devices simultaneously. The number of droplets depends on the size of the droplets, in addition to the area of ​​the first region. For example, if the area is 1. 245×10 10 μm 2 The first region consists of approximately 220,000 liquids with an average droplet diameter of 100 μm. It may contain droplets and 110,000 cells. Area: 1.245 × 10⁻⁶ 10 μm 2 First territory The region contains approximately 432,000 droplets and 216,000 cells, with an average droplet diameter of 80 μm. It is permissible. The area is 1.245 × 10 10 μm 2 The first region has an average droplet diameter of 50 μm. There is a certain approximately 1.2 × 10 6 It may contain droplets and 600,000 cells.

[0022] In some embodiments, the microfluidic chip of the present invention generates oEWOD force. It has an oEWOD structure configured in such a way that it generates an oEWOD force. It may be any structure that can be used.

[0023] In some embodiments, the microfluidic chip of the present invention comprises the following oEWOD structure It includes the structure. That is, the oEWOD structure is A first composite wall comprising a first substrate and a substrate having a thickness in the range of 70 to 250 nm. 1. A transparent conductive layer and a conductor activated by electromagnetic radiation in the wavelength range of 400-1000 nm. A photoactive layer on a layer, the photoactive layer having a thickness in the range of 300 to 1500 nm, and 30 A first composite wall comprising a first dielectric layer on a photoactive layer having a thickness in the range of ~160 nm, , A second composite wall comprising a second substrate and a substrate having a thickness in the range of 70 to 250 nm. A second conductive layer, and optionally, a layer on the second conductive layer with a wavelength of 30-160 nm or 120-16 A second composite wall comprising a second dielectric layer having a thickness in the range of 0 nm, The exposed surfaces of the first and second dielectric layers are spaced less than 180 μm apart, forming microdroplets. A first composite wall and a second composite wall define a microfluidic space adapted to include, A voltage is applied between the ends of the first composite wall and the second composite wall that connect the first and second conductor layers. The A / C source to be supplied, To induce the corresponding virtual electrowetting position on the surface of the first dielectric layer, photoactivation It is adapted to collide with the photoactive layer and has an energy higher than the band gap of the photoactive layer. so, at least one electromagnetic radiation source, By changing the arrangement of the virtual electrowetting positions, the microdroplets are directed to it. We'll create at least one electrowetting path that can be moved along it. The system also includes means for manipulating collision points of electromagnetic radiation on a photoactive layer.

[0024] In some embodiments, the first dielectric layer and the second dielectric layer are composed of a single dielectric material. It may be made of Al2, or a composite of two or more dielectric materials. It can be made from, but is not limited to, O3 and SiO2.

[0025] In some embodiments, the structure may be provided between the first dielectric layer and the second dielectric layer. Good. The structure between the first dielectric layer and the second dielectric layer may be epoxy, polymer, silicon or It may be made of glass, or a mixture or composite thereof, but is not limited to these. Rather, it has walls / surfaces that are straight, angled, curved, or have microstructures. The structure between the dielectric layer and the second dielectric layer is connected to the upper and lower composite walls, sealing the microfluidic deposits. A vise may be constructed to define channels and regions within the device. This structure consists of two parts. It may occupy the gaps between the composite walls.

[0026] In some embodiments, the microfluidic chip of the present invention comprises the following oEWOD structure It includes the structure. That is, the oEWOD structure is A first composite wall comprising a first substrate and a substrate having a thickness in the range of 70 to 250 nm. 1. A transparent conductive layer and a conductive layer activated by electromagnetic radiation in the wavelength range of 400-850 nm. The upper photoactive layer, having a thickness in the range of 300 to 1500 nm, and 20 A first composite wall comprising a first dielectric layer on a photoactive layer having a thickness in the range of 160 nm, A second composite wall comprising a second substrate and a substrate having a thickness in the range of 70 to 250 nm. A second conductive layer, and optionally, a layer on the second conductive layer having a thickness in the range of 20 to 160 nm. A second dielectric layer and a second composite wall comprising, The exposed surfaces of the first and second dielectric layers are spaced 20-180 μm apart, and a microliquid A first composite wall and a second composite wall define a microfluidic space adapted to contain droplets, A voltage is applied between the ends of the first composite wall and the second composite wall that connect the first and second conductor layers. The A / C source to be supplied, To induce the corresponding virtual electrowetting position on the surface of the first dielectric layer, photoactivation It is adapted to collide with the photoactive layer and has an energy higher than the band gap of the photoactive layer. The first electromagnetic radiation source and the second electromagnetic radiation source, By changing the arrangement of the virtual electrowetting positions, the microdroplets are directed to it. We'll create at least one electrowetting path that can be moved along it. The system also includes means for manipulating collision points of electromagnetic radiation on a photoactive layer.

[0027] In some embodiments, the microfluidic chip of the present invention comprises a first composite wall and a second composite It comprises an oEWOD structure including walls. The first composite wall and the second composite wall each consist of a substrate and a conductor. It comprises a layer and a dielectric layer. In addition, the first composite wall has a photoactive layer.

[0028] Each conductive layer has a thickness in the range of 70 to 250 nm and may be transparent. Dielectric layer The photoactive layer may have a thickness in the range of 20 to 160 nm. It is activated by electromagnetic radiation in the wavelength range. The thickness of the photoactive layer is 300-1500 nm. Yes. Furthermore, during use, the exposed surface of the first and second dielectric layers is 20-180 μm. They are positioned at a distance and define the microfluidic space containing minute droplets.

[0029] The chip also has a first composite wall and a second composite wall connected to the first conductor layer and the second conductor layer. The chip also includes an A / C source that supplies voltage between its ends. This also includes the first and second electromagnetic radiation sources, which have even higher energy. The emission source induces the corresponding virtual electrowetting position on the surface of the first dielectric. The chip is adapted to collide with the photoactive layer. Includes a digital micromirror device (DMD). The system changes the arrangement of the virtual electrowetting positions during use, thereby slightly This creates at least one electrowetting pathway along which the small droplets can move. This manipulates the collision points of electromagnetic radiation on the photoactive layer.

[0030] The first and second walls of these structures are transparent, with a microfluidic space sandwiched between them. .

[0031] Appropriately, the first and second substrates are made of mechanically strong materials, such as glass, metal, or e It is made of engineering plastic. In some embodiments, the substrate is to some extent It may have a degree of flexibility. In yet another embodiment, the first substrate and the second substrate are It has a thickness in the range of 100 to 1000 μm. In some embodiments, the first substrate is It is composed of one of silicon, fused silica, and glass. Several embodiments In this configuration, the second substrate is composed of one of fused silica and glass.

[0032] The first conductor layer and the second conductor layer are located on one surface of the first substrate and the second substrate, respectively. In terms of type, the thickness is in the range of 70 to 250 nm, preferably 70 to 150 nm. At least one of the layers is a conductive metal such as indium tin oxide (ITO) or silver. A very thin film, or a transparent conductive material such as a conductive polymer like PEDOT. These layers are made up of a series of discrete structures, such as continuous sheets or wires. Alternatively, the conductive layer may be formed such that electromagnetic radiation is directed between the gaps in the mesh. A mesh made of conductive material may also be used.

[0033] The photoactive layer generates localized charge regions in response to stimulation from the second electromagnetic radiation source. It is appropriately constructed from available semiconductor materials. For example, a thickness in the range of 300 to 1500 nm. Examples include a hydrogenated amorphous silicon layer having a light The active layer is activated by the use of visible light. Photoactive layer in the case of the first wall and the second wall The optional conductive layer in this case is coated with a dielectric layer that is typically 20-160 nm thick. The dielectric properties of this layer preferably have a high dielectric strength of >10^7 V / m and > It contains a dielectric constant of 3. Preferably, it is as thin as possible in accordance with avoiding dielectric breakdown. In some embodiments, the dielectric layer is alumina, silica, hafnia, or a thin non-conductive layer. Selected from electrolytic polymer films.

[0034] In another embodiment of these structures, at least the first dielectric layer, preferably both, is used to prevent fouling. By coating in layers, the desired microdroplet / carrier fluid / surface contact angle can be adjusted to various virtual elements. It helps to establish the electrode position with a microdroplet, and additionally, the microdroplet on the tip As it moves along, the contents of the tiny droplets adhere to the surface, preventing them from decreasing. If the second wall does not contain the second dielectric layer, the second antifouling layer is applied directly onto the second conductor layer. That's good too.

[0035] For optimal performance, the antifouling layer was measured at 250°C as an air-liquid-surface three-point interface. In this case, establish the microdroplet / carrier fluid / surface contact angle, which should be in the range of 50-180°C. This should help. In some embodiments, these layers are less than 10 nm thick. It has a monolayer, and is typically a single layer. In other cases, these layers are made of methyl methacrylate. Acrylates polymers, or derivatives thereof substituted with hydrophilic groups, for example, It consists of lucoxysilyl. Either or both of the antifouling layers are used to ensure optimal performance. It is hydrophobic. In some embodiments, in order to provide chemically compatible crosslinking, 2 A silica interstitial layer with a thickness of less than 0 nm is interposed between the antifouling coating and the dielectric layer. It is possible.

[0036] The first dielectric layer and the second dielectric layer, and therefore the first wall and the second wall, define the microfluidic space. The microfluidic space has a width of at least 10 μm, preferably in the range of 20 to 180 μm. And, within it, tiny droplets are contained. Preferably, before they are contained, the tiny droplets themselves are It has an intrinsic diameter that is more than 10%, and preferably more than 20%, than the width of the microdroplet space. Therefore, upon entering the chip, the microdroplets are compressed, resulting in, for example, better microdroplet aggregation ability. Through this, electrowetting performance is improved. In some embodiments, the The first dielectric layer and the second dielectric layer are coated with a hydrophobic coating such as fluorosilane.

[0037] In another embodiment, the microfluidic space is maintained with the first and second walls separated by a predetermined amount. Includes one or more spacers for light patterning. Therefore, the resulting intermediate resist layer contains beads, pillars, or bumps. Alternatively, spacers can be made using deposited materials such as silicon oxide or silicon nitride. Alternatively, a flexible plastic film may be used with or without an adhesive coating. A spacer layer can be formed using a film layer containing a film. Using a S-shape, narrow channels, tapering channels, defined by pillar lines. Alternatively, it can form a partially enclosed channel. With careful design, this can be achieved. These spacers can be used to help deform the microdroplets, followed by the splitting of the microdroplets. And effects can be performed on deformed microdroplets. Similarly, these Spacers are used to physically separate zones on the tip and prevent cross-contamination between droplet clusters. This can promote the flow of droplets in the correct direction when a load is applied to the tip under hydraulic pressure. ru.

[0038] The first and second walls are biased using an A / C power source attached to the conductive layer. This provides a voltage potential difference between them. Ideally, this is in the range of 10 to 50 volts. Our oEWOD structure is typically used in conjunction with a second electromagnetic radiation source. The radiation source has a wavelength in the range of 400-850 nm, preferably 660 nm, and the photoactive layer is van It has energy exceeding the gap. Appropriately, the photoactive layer is used with respect to the incident intensity of the irradiation. The degree is 0.01~0.2Wcm -2 Activated at the virtual electrowetting electrode position .

[0039] When the electromagnetic radiation source is pixelated, it is illuminated by light from an LED or other lamp. Using reflective screens such as digital micromirror devices (DMDs), directly Alternatively, an electromagnetic radiation source is supplied appropriately. This allows for virtual electrowetting. The highly complex pattern of the electrode positions is rapidly created on the first dielectric layer and then destroyed. This makes it possible to use tightly controlled electrowetting forces. This makes it possible to precisely manipulate microdroplets along essentially any virtual path. Such electrowetting paths are virtual electrowetting on the first dielectric layer It can be considered as being constructed from a continuum of electrode positions.

[0040] The collision point of the electromagnetic radiation source with the photoactive layer can be any convenient shape, including conventional circular or annular shapes. This can be done. In some embodiments, the form of these points corresponds to the pic. Determined by the cellular structure, or alternatively, by the form of the microdroplets once they enter the microfluidic space. It corresponds to, completely or partially. In one embodiment, the collision point, and therefore, the electric The position of the trowetting electrode may be crescent-shaped, and the intended movement of the microdroplets may be They may be oriented in the direction. Appropriately, the electrowetting electrode position itself is on the first wall. Smaller than the surface of the tiny droplets adhering to the surface, the contact line formed between the droplet and the surface dielectric crosses the surface. This provides the maximum possible electric field strength gradient.

[0041] In some embodiments of the oEWOD structure, the second wall emits the same or different electromagnetic radiation. The source can also induce the position of a virtual electrowetting electrode on the second dielectric layer. It also includes a photoactive layer. The addition of the second dielectric layer causes microdroplets to flow from the top to the bottom of the structure. Edge transition and application of more electrowetting force to each microdroplet This makes it possible.

[0042] The first and second dielectric layers may be composed of a single dielectric material, or two or more dielectric materials. It may be a composite of electrical materials. The dielectric layer may be made from Al2O3 and SiO2. However, it is not limited to these.

[0043] The first and second dielectric layers minimize the adverse effects of pinhole defects. This makes it easy to simultaneously manipulate thousands of tiny droplets over a relatively large area. The dielectric layer always has sparse pinhole defects, and conductivity is present in small, isolated regions. Yes. Pinhole defects can trap droplets and render them immobile. The impact of this... This is more serious when using droplets of a conductive medium, such as a buffer solution. The first and second dielectric layers can operate below the dielectric breakdown voltage, and the conductive path By minimizing the possibility of forming any single pinhole defect, The effect of the rifle defect can be ruled out. This pinning is achieved by the presence of the second dielectric layer. Hall relaxation features are key to enabling the simultaneous manipulation of thousands of droplets over a relatively large area. Yes. In some embodiments, the device is 50 cm 2 Approximately 50 across a region exceeding [a certain area] It is possible to manipulate ,000 droplets simultaneously.

[0044] In some embodiments, optically mediated electrowetting is dielectric Applying a voltage below the dielectric breakdown voltage of the layer across the first and second dielectric layers. This can be achieved by optically mediated electro Lighting can be achieved using low power supplies such as LEDs. In some embodiments... In this case, the electrowetting mediated by optics is 0.01 W / cm². 2 Power This can be achieved with a lighting source that allows the device to operate below the dielectric breakdown voltage. This eliminates the adverse effects of dielectric pinholes. Low power allows for non-conductive In addition to microdroplets, this enables the manipulation and control of conductive droplets.

[0045] In some embodiments, the device is used to prevent damage from high current. Conductive microdroplets formed from an ion buffer containing biomolecules capable of handling and It is controllable. A low voltage applied across the two dielectric layers causes the destructive ionization of the conductive droplet. It prevents ionization and protects biomolecules from destruction.

[0046] The structure may be provided between the first dielectric layer and the second dielectric layer. The structure between the layers is epoxy, polymer, silicon, or glass, or a mixture thereof. They may be made of composite materials, but are not limited to these, and may be linear, angular, etc. The structure between the first dielectric layer and the second dielectric layer has curved or microstructured walls / surfaces. It connects to the upper and lower composite walls to create a sealed microfluidic device, and inside the device Channels and regions may be defined. This structure may occupy the gap between the two composite walls. Good. Alternatively or additionally, the conductor and dielectric can be deposited on a molded substrate that already has walls. You may do so.

[0047] Some aspects of the methods and apparatus of the present invention involve micro-electrophoresis or optical tweezers. Electrowetting devices such as devices configured to manipulate particles It is suitable for application to external optical activation devices. In such devices, cells Alternatively, the particles are manipulated and examined using functionally identical optical instruments, and a virtual optical induction is performed. Generates an electrophoretic gradient. Microparticles as defined herein include living cells, polystyrene, and Microbeads, hydrogels, and magnetic microbeads made from latex-containing materials Alternatively, it may refer to particles such as colloids. Dielectrophoresis and optical tweezers mechanisms are applicable to those skilled in the art. It is well known and can be easily implemented by those skilled in the art.

[0048] In some embodiments, the microdroplets contain biomaterial, one or more cells, and It may contain one or more beads. In some embodiments, the microdroplets are , biological cells, cell culture media, chemical compounds or compositions, drugs, enzymes, and their surface of any choice It may include beads or microspheres having materials bound together by . More specifically, the cells are It can be a mammal, bacterium, fungus, yeast, macrophage, hybridoma, C HO, Jurkat, CAMA, HeLa, B cells, T cells, MCF-7, MDAMB- 231, may be selected from, but is not limited to, Escherichia coli or Salmonella. The chemicals contained within the microdroplets include enzymes, assay reagents, antibodies, antigens, drugs, and antibiotics. It can be used as a dissolving reagent, surfactant, dye, or cell staining agent. Other biological or chemical substances that may be mixed include DNA oligos, nucleotides, loads. Or unloaded beads / microspheres, fluorescent reporters, nanoparticles, nanowires or magnets Contains porosity particles.

[0049] In some embodiments, the constriction may be a physical element such as a physical barrier. stomach.

[0050] In some embodiments, the constricting means may include an opening or gap. Microdroplets from one region may enter the second region, and vice versa, through the gap. The opening is The microdroplet must have a width sufficient to pass through the first region and enter the second region. In some embodiments, the width of the opening may be between 20 and 200 microns. i. In some embodiments, the width of the opening is 20, 40, 60, 80, 100, 1 It may be larger than 20, 140, 160, or 180 microns. Some embodiments In this case, the width of the opening is 200, 180, 160, 140, 120, 100, 80, 6 The opening may be less than 0, 40, or 30 microns. In some embodiments, The width may be between 20 and 400 microns. In some embodiments, the opening The width of the section is 20, 50, 100, 150, 200, 250, 300, or 350 microns. It may be larger than that. In some embodiments, the width of the opening is 400, 350, It may be less than 300, 250, 200, 150, 100, 50, or 30 microns. .

[0051] The term "constricting means" as disclosed in this invention, unless otherwise specified, is used herein. The term "constriction" refers to any part that makes it possible to separate the first and second regions. It means structure or arrangement. The constricted area is a wall or This may be a physical element such as a barrier. Alternatively or additionally, the constricted portion may be a sheath liquid flow. Alternatively, a semipermeable membrane may be used.

[0052] In some embodiments, the constricted portion may be a semipermeable membrane. The semipermeable membrane is molecular or It may be provided to enable selective diffusion of ions. In some embodiments The semipermeable membrane may be non-porous.

[0053] In some embodiments, the constriction may be a flow of sheath fluid. Disclosed in the present invention Unless otherwise specified, the terms “sheath fluid” or “sheath flow” refer to fluids that are not mixed. This refers to at least two fluids with sufficiently different densities or velocities.

[0054] In some embodiments, the geometry of the second region is substantially crescent-shaped. It may also be a crescent shape or a horseshoe shape configuration, with an inlet port and an outlet port in the second region. This allows the ports to be manufactured in extremely close proximity inside the device, which can be advantageous in some cases. This configuration maximizes the usable space inside the microfluidic chip. Furthermore, the crescent shape also reduces the burden of manufacturing the device and lowers manufacturing costs. This has the additional advantage of having an inlet port and outlet port in the second region. The distance between the mouth port may be 1500 μm. Alternatively, the geometries of the second region The channel may be semicircular, square, rectangular, or curved. It may also be a bird. In some embodiments, the second area is required on the chip. To accommodate other possible features or structures of microfluidics, straight, curved, Alternatively, it may have a meandering geometry. In some embodiments, the second region The ometrium may be any suitable shape or configuration.

[0055] The geometry of the second region may have a channel width between 10 and 1000 microns. The second area may have channels of a fixed width or a variable width. Several implementations In this configuration, the channel width may narrow towards the inlet or outlet port. This reduces the possibility of low flow rate regions occurring where droplets may clog, and allows droplets to form microfluidic chips. It can also reduce the time it takes to get out of the tub.

[0056] In some embodiments, the width of the crescent-shaped channel is 10, 20, 40, 60 , 80, 100, 120, 140, 160, 180, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, It may be larger than 900 or 950 microns. In some embodiments, crescent shape. The channel shapes are 1000, 950, 900, 850, 700, 750, 700, 65 0, 600, 550, 500, 450, 400, 350, 300, 250, 200, 18 0, 160, 140, 120, 100, 80, 60, 50, 40, 30 or 20 microns It may have a width of less than n.

[0057] In some embodiments, the second region may further comprise a plurality of channels. The channel receives a microdroplet from the first region and sends the microdroplet to the outlet port of the microfluidic chip. It may be configured to move to the destination.

[0058] In some embodiments, the second region may have 1 to 1000 channels. In some embodiments, the second region is 1, 10, 50, 100, 150, 200, 2 50, 300, 350, 400, 450, 500, 550, 600, 650, 700, 7 It may have more than 50, 800, 850, 900, or 950 channels. In this embodiment, the second region is 1000, 950, 900, 850, 750, 700 , 650, 600, 550, 450, 400, 350, 300, 250, 200, 150 It may have 100, 50, or fewer than 10 channels.

[0059] In some embodiments, each of the multiple channels in the second region is substantially three It may have a sun-moon shaped geometry. In some embodiments, in the second region Each of the multiple channels may have a horseshoe-shaped configuration. In some embodiments, Each of the multiple channels in the second region may have a semicircular geometry. Alternatively, each channel may be a square, rectangular, or curved geometry. In some embodiments, each of the multiple channels in the second region is required on the chip To accommodate other microfluidic features or structures that may be considered to be straight, curved It may have a curved or meandering geometry. In some embodiments, the second region Each of the multiple channels in the region may have any suitable shape or configuration. Crescent shape Alternatively, the horseshoe-shaped configuration allows the inlet and outlet ports of the second region to be extremely close together inside the device. This configuration allows for manufacturing in close proximity, which can be advantageous. This maximizes the usable space inside the unit. Furthermore, the crescent-shaped configuration also allows for... It has the added benefit of reducing the burden of manufacturing devices and lowering manufacturing costs.

[0060] In some embodiments, the multiple channels in the second region are arranged in parallel. Alternatively, in some embodiments, the multiple channels in the second region are arranged in series. It may also be used.

[0061] In some embodiments, multiple channels in the second region are used to control the flow of microdroplets. Sorting may be made easier. In some embodiments, multiple channels are connected to a single exit. They may be bonded together. In some embodiments, the target microdroplets and unrelated The microdroplets found to be such may be dispensed from the device through the same outlet. In one embodiment, the multiple channels in the second region are multiple outputs in the second region. It may be guided to the mouth. Multiple channels and multiple outlets allow multiple microdroplets to enter the microfluidic system. It may be configured to dispense multiple droplets simultaneously from the vise. Dispensing minimizes the time required to dispense microdroplets from the device. This maximizes the device's processing power.

[0062] In some embodiments, microdroplets are released from a microfluidic device in any desired order. It may be dispensed. In some embodiments, the microdroplets are loaded into the microfluidic device. The devices may dispense the contents in the same order as they were dispensed. In some embodiments, The microdroplets are dispensed from the device in an order different from the order in which they were loaded into the fluid device. That's fine.

[0063] In some embodiments, the device extends from the inlet port to the outlet port of the microfluidic chip. The system may further include means for controlling the flow of carrier fluid through the microfluidic chip up to the terminal.

[0064] In some embodiments, means for controlling the flow rate of the carrier fluid include valves and / or It may be a pump. Just as an example, the pump could be a syringe pump or a high-pressure pump. It may be present. The valve may be a 2-port 2-way valve or a 3-port switching valve.

[0065] In some embodiments, the means for controlling the flow is a syringe pump or a high-pressure pump. These pumps may be software-controlled pump sources such as microfluidic chips. It may be connectable to one of the inlet ports. In combination with one or more switching valves. The pump may be connected to multiple ports on the microfluidic chip. This allows one to Alternatively, multiple ports can receive the flow. Meanwhile, other ports are sealed. By having a software-controlled pump, the pump source can be controlled without requiring manual intervention. It can be automatically controlled and turned on or off. Additionally or alternatively, the valve and / or the pump can be controlled manually. Furthermore, it can be used to control the flow of the carrier fluid. The pump and / or valve used will deliver microfluidic fluid from the inlet port to the outlet port of the microfluidic chip. It provides a constant flow rate throughout the entire small fluid chip.

[0066] In some embodiments, means for controlling the flow, such as valves and / or pumps, The conduit may be configured to connect to the outlet port of the microfluidic chip. This can be a tube with an inner diameter of 20 to 500 microns. In some embodiments... In this case, the conduits are 20, 50, 100, 150, 200, 250, 300, 350, and 40. It may have an inner diameter greater than 0 or 350 microns. In some embodiments, The conduits are 500, 450, 400, 350, 300, 250, 200, 150, and 50. Alternatively, it may have a diameter of less than 20 microns.

[0067] In some embodiments, the valve is a 2-port 2-way valve, a 4-port 2-way valve, and / or This may be a 6-port 2-way valve. The valve may also have an additional "closed" position. Therefore, the outlet port of the microfluidic chip is sealed to prevent fluid from flowing. To achieve similar results, multiple valves are connected in sequence or network. It's okay.

[0068] By having a 4-port, 2-way valve, the valve controls the microfluidic tip while droplets are being dispensed. It can seal the pores, reducing the possibility of unwanted droplet movement inside the microfluidic chip. Reduce. A 4-port 2-way valve allows droplets to pass through the valve once before reaching a higher flow rate. Dispensing may be accelerated by using [a specific method]. In addition, the pressure inside the tip can be controlled more effectively. There is a possibility that this will happen.

[0069] By providing a 6-port, 2-way valve, only the desired droplets are captured in the capture loop. By capturing them, bubbles and / or excess droplets are significantly reduced, or made easier to capture. There is an additional advantage in that it removes it. In addition, by using a 6-port 2-way valve, sample This allows the loop to be introduced into the conduit, and only a small amount of fluid is dispensed from within the tip. This may be done so that only a small amount of immiscible carrier medium is dispensed along with the droplet. This makes it possible to use an aqueous medium as the carrier phase for dispensing, thus improving the dispensing process. The amount of immiscible carrier medium required is reduced.

[0070] By providing a 4-port 2-way valve, a bypass route may be provided, once Once the droplets are removed from the tip through the valve, the flow is directed from the pump directly into the dispensing conduit containing the droplets. The contact can be rerouted. Further movement of the droplet requires fluid to pass through the tip. No. This will result in unwanted droplets or other materials being dispensed from the tip into the body. The possibility of introducing it into the product is reduced. In addition, this reduces the possibility of disturbance to the contents of the chip. The quality may also be reduced. Furthermore, as a result, the contents of the tip are subjected to a high flow rate. The time spent exposed to the resulting higher pressure can be reduced. The purpose is to enable the immediate initiation of further oEWOD operations within the second region, and thereafter This can reduce the time required for dispensing operations.

[0071] Additionally or alternatively, an 8-port 2-way valve or a 10-port 2-way valve may be provided. An 8-port 2-way valve or a 10-port 2-way valve can further accelerate the dispensing process. Alternatively, it may be possible to incorporate a second sampling loop into the conduit.

[0072] In some embodiments, a multi-port switching valve is provided. As disclosed in the details, it can be used in combination with any other valve to further the dispensing process It can be duplicated.

[0073] The device according to the present invention has a valve connected to the outlet port of the second region of a microfluidic chip. It further comprises a controller configured to control means of flow such as a pump and / or The controller may be software on a computer or microprocessor. It can also be an application.

[0074] In some embodiments, the carrier fluid from the microdroplets exits the microfluidic device The controller is activated to open the valve, allowing the water to exit through the port. You may replace it, or you may switch on the pump. The pump provides a specific flow rate. The pump may be controlled to supply or maintain a constant flow rate. It may be controlled to do so. One or more valves are controlled to a specific inlet port of the microfluidic device. Fluid flow, so as to enter and exit a conduit, and / or along a specific connected conduit. It may be controlled to lead to this.

[0075] In some embodiments, the device disclosed in the present invention detects a microfluidic signal. It may also be possible to further incorporate a detection system that detects from the microdroplets dispensed from the tip's exit port. i. In some embodiments, all or part of the inside or near the connected conduit. A precisely positioned sensor or detection module can detect the specific location of the connected conduit. A detection system is used to detect the presence or absence of dispensed microdroplets in the area. That's good too.

[0076] The detection system may include sensors or detectors. In some embodiments, The sensor or detector can be an optical sensor or an electrical detector. An example of an optical sensor is Light source, lens device and photodiode, or phototransistor, or lens and cameras, but not limited to these. Electrical sensors or detectors Examples include, but are not limited to, capacitive detectors or impedance detectors. It will not be done.

[0077] In some embodiments, the controller controls each of the channels in the second region. It is configured to control the flow of microdroplets, or the flow of each individual microdroplet simultaneously. It may also be the case that the simultaneous flow of microdroplets in multiple channels in the second region is the microdroplet The time required to dispense from the device can be minimized. Several implementations Morphologically, the controller controls the microdroplets in each channel in the second region. It may be configured to continuously control the flow. Through multiple channels in the second region The continuous flow of microdroplets facilitates sorting of the microdroplets before dispensing from the device. It can be done.

[0078] In some embodiments, multiple microdroplets are simultaneously delivered to the outlet port of a microfluidic chip. It can be moved to a microfluidic. In some embodiments, multiple microdroplets are moved to a microfluidic. It can be dispensed simultaneously from the tip.

[0079] In some embodiments, the device of the present invention has an inlet port or outlet port in a first region. The port and a valve provided at the inlet or outlet port of the first region may be further provided. i. In some embodiments, the device has an inlet port or an outlet port in the first region. It may further include a valve connected by the first region. In order to prevent flow in the first region It is advantageous to provide valves at the inlet and / or outlet ports. This ensures that the flow in the second region does not interfere with the manipulation or storage of droplets within the first region. This is guaranteed.

[0080] In some embodiments, the device comprises a reader module with analog circuitry. Further features may be provided. The reader module receives signals generated from the sensor or detection module. It is configured to read the code and send it to the controller. Then the controller, The valve may be further configured to be positioned in the open position so that minute droplets are dispensed. In one embodiment, the device receives signals generated from a sensor or detection module. It may also include a reader module configured to read and send data to the controller. Furthermore, the controller will position the valve in the open position so that minute droplets are dispensed. It is further composed of a leader module, which is a controller such as a microcontroller. It is also acceptable. The valve controls the direction of flow in the well plate and / or dispensing head. It may be used for the purpose of... In some embodiments, the flow is directed to a waste container or... It may be directed towards the channel. However, if a droplet is detected, the droplet will be directed towards the well plate. The valve is switched so that it is directed towards another dispensing receptacle.

[0081] In some embodiments, the apparatus of the present invention may further include a receptacle. The septacle can be configured to receive dispensed microdroplets. In embodiments, the apparatus of the present invention may further include a receptacle. The receptacle is It can be configured to receive dispensed microdroplets.

[0082] In some embodiments, the receptacle is a multiwell plate, a PCR tube It is a tubing or microcentrifuge tube. The receptacle is a 96 or 384 multiwell plate. It can be a multiwell plate such as a t. Alternatively, the receptacle is PC Microcentrifuge tubes such as R-tubes or Eppendorf tubes, or other suitable tubes. It can be used as a container.

[0083] In some embodiments, a multiwell plate is placed on a multi-axis motion-controlled stage. It may be mounted. The multi-axis motion-controlled stage is positioned so that the target well is below the valve outlet port. The multiwell plate can be configured to move to a first position. The multi-axis motion-controlled stage may also be an X, Y, and Z-axis motion-controlled stage. In one embodiment, the multiwell plate is placed on a multi-axis motion-controlled stage. It is also possible that the multi-axis motion-controlled stage has a target well located at the outlet port of the microfluidic chip. The multiwell plate is moved to the first position so that it is positioned below the valve. It is possible.

[0084] Alternatively, the valve or dispensing head can be used when the well plate is stationary and the dispensing head is in the well plate. It may be mounted on a motion-controlled stage so as to move along the surface. Alternatively, Both the gel plate and the dispensing head may be mounted on a motion-controlled stage.

[0085] In some embodiments, the 6, 8, or 10 port valve is switched to the open position to access the conduit. The sensor is placed inside the conduit, that is, to trap droplets in the sample loop. It can be placed within the sample loop. It triggers the droplets dispensed into the receptacle. Therefore, a second sensor is provided to detect droplets in the dispensing tube near the dispensing head. This can be done. A sampling loop is used, and droplets are dispensed using an aqueous medium. In an embodiment that enables this, the sensor captures in the sampling loop Furthermore, the presence of a plug of an immiscible carrier fluid, which will likely contain minute droplets, is detected. Ro.

[0086] In some embodiments, each well may be pre-filled with a certain amount of cell culture medium. The cell culture medium may contain EMEM, DMEM, RPMI, K12, and Hams, however These are not the only options.

[0087] In some embodiments, each well contains one of the following: buffer, water, or It may be pre-filled with one or more fixed amounts of oil. In some embodiments, The buffer may be a solubilating buffer. In some embodiments, the buffer or water or The oil may contain components or requirements used in subsequent assays. For example, P If CR or qPCR follows, the requirement may include primers or appropriate controls. Cut.

[0088] In some embodiments, during the dispensing procedure disclosed herein, the end of the outer tube The ends of conduits, such as those in the well, may be lowered below the surface of the pre-filled volume within the well.

[0089] The dispensing system is designed to reduce the possibility of cross-contamination by incorporating conduits, valves, dispensing heads, and valves. The formulation may include additional ingredients or processes for cleaning the tube between injections.

[0090] In another embodiment of the present invention, a method for dispensing one or more microdroplets is provided. The method includes the following steps: A microfluidic chip is provided comprising a first region and a second region separated by a constricted portion. The steps, The process includes the step of transferring a microdroplet from a first region to a second region, wherein the microdroplet is in the first region In the first region, the first region is dispersed in the carrier fluid at a first flow rate, and the second region is separated from the first region by a constricting means. It receives microdroplets and delivers these microdroplets to the microfluidic chip at a higher carrier fluid flow rate. It is configured to move to the exit port, The second region is the application of optically mediated electrowetting (oEWOD) forces. Therefore, it is configured to receive the minute droplet from the first region through the constricted portion. The second flow rate in the second region is higher than the first flow rate in the first region.

[0091] In some embodiments, a method for dispensing one or more microdroplets is provided. The method includes the following steps: The present invention provides a microfluidic chip comprising a first region and a second region separated by a constricting means. The steps to take, The process includes the step of transferring a microdroplet from a first region to a second region, wherein the second region is a constricted area. The system receives microdroplets from the first region via a step, and the microdroplets are then transported to a higher carrier fluid flow rate. In a method configured to move the microfluidic chip to the exit port, The second region is the application of optically mediated electrowetting (oEWOD) forces. Therefore, it is configured to receive the minute droplet from the first region via the constriction means.

[0092] The method of the present invention uses a controller to carry the carrier through the outlet port of the microfluidic chip The step further includes operating a pump and / or valve to control the flow of fluid. This is also acceptable. In some embodiments, the method uses a controller to control the microfluidics Further steps include activating means to control the flow of carrier fluid through the outlet port of the piping. It may be included.

[0093] In some embodiments, means for controlling the flow of carrier fluid are microscopically controlled by a conduit. It may be connected to the outlet port of a small fluid chip.

[0094] In some embodiments, the means for controlling the flow of the carrier fluid is a pump and / Alternatively, a lawyer would also be acceptable.

[0095] In some embodiments, the pump operates by moving a fixed volume of fluid to a microfluidic chip. The steps may include moving through the pipe and conduit. The conduit is a tube, i.e., an external It can be a side tube. The outer tube can be made of plastic. In some embodiments, the outer tube is transparent. The outer tube is made of a fluoropolymer. Preferably, the outer tube is made of fluorinated ester. It is ethylene propylene (FEP). As a result, the operator and sensor will see the droplets on the outside. You can see it moving inside the tube. The tube may have any length. However, it can be a tube between 10 and 1000 mm in length. For example, The outer tube is designed to extend to the opposite side of the well plate (130mm x 85mm). It can be made into a 200mm tube.

[0096] The amount of fluid supplied to move through the microfluidic chip and conduit is 1 to 10 It is μl. In some embodiments, the fixed volume of liquid is 2, 3, 4, 5. It can be more than 6, 7, 8 or 9 μl. In some embodiments, the liquid fixed volume may be less than 10, 9, 8, 7, 6, 5, 4, 3 or 2 μl as well.

[0097] Preferably, the fixed amount of liquid is 7 μl. The volume of 7 μl is significantly less than the volume of the target well plate but can be significantly more than the volume of the conduit or fluid path that has to be flushed.

[0098] In some embodiments, the method may further include a receptacle. The receptacle can be a multi-well plate or can be a PCR tube.

[0099] The method of the present invention comprises the step of placing a multi-well plate on a multi-axis motion control stage wherein the multi-axis motion control stage is configured to move the multi-well plate to a target well using a controller and the target well is located under a valve provided at the outlet port of the microfluidic chip, and may further include the step of In some embodiments, the method comprises the step of placing a multi-well plate on a multi-axis motion control stage wherein the multi-axis motion control stage is configured to move the multi-well plate to a target well using a controller and the target well is located under the outlet port of the valve, and may further include the step of

[0100] In some embodiments, the method may further include the step of switching the valve to an open position so that micro-droplets are dispensed onto the multi-well plate.

[0101] In some embodiments, the method may further include the step of recording a target well using a controller. By recording the target well well using a controller, an operator or user can know which wells contain the desired droplets, such as droplets containing cells. In some cases, there may be an assay in which the target well can be selected for the dispensed droplets. In some embodiments, the method may further include the step of well selecting a target well using a controller so that the desired droplets can be dispensed into the target well. In some embodiments, a software function is used to assign a unique identifier to a droplet and record metadata regarding the operations performed on that droplet. This metadata can include a record of the target well into which the droplet was dispensed. If the droplet is split into two droplets, the metadata can include a record of the target recovery well for one of the dispensed daughter droplets, as well as the unique identifier of the other daughter droplet retained on the chip.

[0102] In some embodiments, the method may include the step of optically inspecting the droplets using a brightfield microscope, a fluorescence microscope, or a darkfield microscope. The method may include performing image analysis to classify the droplets and then selecting target wells for the dispensed droplets based on those classifications. In some embodiments, the method may include the step of providing a detection module or near a conduit.

[0103] In some embodiments, a software function is used to assign a unique identifier to a droplet and record metadata regarding the operations performed on that droplet. This metadata can include a record of the target well into which the droplet was dispensed. If the droplet is split into two droplets, the metadata can include a record of the target recovery well for one of the dispensed daughter droplets, as well as the unique identifier of the other daughter droplet retained on the chip. In some embodiments, a software function is used to assign a unique identifier to a droplet and record metadata regarding the operations performed on that droplet. This metadata can include a record of the target well into which the droplet was dispensed. If the droplet is split into two droplets, the metadata can include a record of the target recovery well for one of the dispensed daughter droplets, as well as the unique identifier of the other daughter droplet retained on the chip. This metadata can include a record of the target well into which the droplet was dispensed. If the droplet is split into two droplets, the metadata can include a record of the target recovery well for one of the dispensed daughter droplets, as well as the unique identifier of the other daughter droplet retained on the chip. In some embodiments, a software function is used to assign a unique identifier to a droplet and record metadata regarding the operations performed on that droplet. This metadata can include a record of the target well into which the droplet was dispensed. If the droplet is split into two droplets, the metadata can include a record of the target recovery well for one of the dispensed daughter droplets, as well as the unique identifier of the other daughter droplet retained on the chip. This metadata can include a record of the target well into which the droplet was dispensed. If the droplet is split into two droplets, the metadata can include a record of the target recovery well for one of the dispensed daughter droplets, as well as the unique identifier of the other daughter droplet retained on the chip.

[0104] In some embodiments, the method may include the step of optically inspecting the droplets using a brightfield microscope, a fluorescence microscope, or a darkfield microscope. The method may include performing image analysis to classify the droplets and then selecting target wells for the dispensed droplets based on those classifications. In some embodiments, the method may include the step of optically inspecting the droplets using a brightfield microscope, a fluorescence microscope, or a darkfield microscope. The method may include performing image analysis to classify the droplets and then selecting target wells for the dispensed droplets based on those classifications. In some embodiments, the method may include the step of optically inspecting the droplets using a brightfield microscope, a fluorescence microscope, or a darkfield microscope. The method may include performing image analysis to classify the droplets and then selecting target wells for the dispensed droplets based on those classifications. In some embodiments, the method may include the step of optically inspecting the droplets using a brightfield microscope, a fluorescence microscope, or a darkfield microscope. The method may include performing image analysis to classify the droplets and then selecting target wells for the dispensed droplets based on those classifications.

[0105] In some embodiments, the method may include the step of providing a detection module or near a conduit. The step may further include generating a signal using a sensor. In some embodiments, The method further includes the step of generating a signal using a detection module or sensor. That's fine.

[0106] In some embodiments, the method involves detecting the generated signal in a detection module or sensor. The steps include detecting from and sending the generated signal to the controller, and further It may include. Then, the controller opens the valve so that minute droplets are dispensed. It is further configured to switch. In some embodiments, the method generates the signal Steps include detecting a number from a detection module or sensor and controlling the generated signal. The controller may further include the step of sending to R. The system is further configured to switch the valve to the open position so that the contents are dispensed.

[0107] In some embodiments, the method may further include the following steps: A step of using the controller to deactivate the pump, The steps include switching the valve to the closed position using the controller, Using the controller, the valve outlet port is directed to a further target well different from the first target well. A step positioned above the shell, A step of restarting the pump using a controller, wherein the pump is fixed A step configured to move a certain amount of fluid through a microfluidic tip and a conduit. , A step of switching a valve to the open position using a controller, wherein the fluid is multi-way The step of dispensing into a plate, The steps involve recording additional target wells using a controller.

[0108] According to a further aspect of the present invention, there is provided an apparatus for dispensing one or more microdroplets. . The apparatus comprises the following. That is, a microfluidic chip as described herein, including a second region configured to transfer microdroplets dispersed in a carrier fluid to an outlet port of the microfluidic chip, a microfluidic chip, a pump configured to control the flow of the carrier fluid through the microfluidic chip from an inlet port to an outlet port of the microfluidic chip, a conduit connected to the outlet port of the microfluidic chip, the conduit being configured to receive the microdroplets once dispensed from the chip, a sensor disposed in the vicinity of the conduit and configured to generate a signal, a reader module configured to read the generated signal from the sensor and transmit it to a controller, wherein the controller is configured to control a valve and / or a pump connected to the outlet port of the microfluidic chip, and in response to the signal generated by the sensor, the controller is configured to switch the valve to a position where the microdroplets are dispensed from the apparatus, or the controller is configured to switch the valve to a position where the microdroplets are dispensed onto a receptacle. The apparatus comprises:

[0109] Here, the present invention will be described in further detail by way of example only, with reference to the accompanying drawings.

Brief Description of the Drawings

[0110] [Figure 1] It is a diagram of a microfluidic device for dispensing one or more microdroplets disclosed in the present invention. [Figure 2A] It is a diagram showing a chip loading and dispensing sequence disclosed in the present invention.​​​​ [Figure 2B] This figure shows the tip loading and dispensing sequences disclosed in the present invention. [Figure 2C] This figure shows the tip loading and dispensing sequences disclosed in the present invention. [Figure 3A] Figures 2A-2C show the droplet dispensing procedure and detection. [Figure 3B] Figures 2A-2C show the droplet dispensing procedure and detection. [Figure 4A] This diagram shows the operation of liquid droplets inside a microfluidic device. [Figure 4B] This diagram shows the operation of liquid droplets inside a microfluidic device. [Figure 4C] This diagram shows the operation of liquid droplets inside a microfluidic device. [Figure 4D] This diagram shows the operation of liquid droplets inside a microfluidic device. [Figure 5A] This figure shows droplets being dispensed in a multiphase flow. [Figure 5B] This figure shows droplets being dispensed in a multiphase flow. [Figure 6] This is a diagram of a device or system for dispensing liquid droplets. [Modes for carrying out the invention]

[0111] Referring to Figure 1, a micro... A fluid device 10 is provided. The enclosed volume 12 is comprised of a first region 14 and a second region 16 Includes. The first region 14 is where droplets are stored, handled, and / or manipulated (Figure 1) It may be a large area as shown. The first area 14 and the second area 16 are walls or It is separated by a narrowing means 18 such as a barrier. The wall or barrier 18 as shown in Figure 1 is It has a gap 20 wide enough for the droplet to pass through and enter the second region 16. It contains the target cells. A droplet is selected, and then an optoelectrowetting (oEWOD) force is applied. It is moved through the gap 20. This device also has several ports 22, 24 It has 26 and 28. These can be opened independently using connected valves, or It can be sealed.

[0112] The first region 14 contains one or more microdroplets dispersed in the carrier fluid as low-carrier fluids. It is adapted to receive and operate at fluid flow rates. In some cases, low carrier liquid flow rates are used. This is zero in the first region 14. This makes it easy to manipulate and handle droplets. This is possible. If the flow rate is too high in the first region 14, the flow rate will be such that the droplets are held in place. It overwhelms the power of manipulating liquid droplets or other liquids.

[0113] The flow rate in the first region can be in the range of 0 to 20 μL / min, or 0, 2, 4 It may exceed 6, 8, 10, 12, 14, 16, or 18 μL / min. The flow rate in the first region is 20, 18, 16, 14, 12, 10, 8, 6, 4, or 2 μL / Less than a minute.

[0114] The second region 16 may have two different flow rates. During the ND-by mode, the flow rate in the second region can be set between 0 and 20 μL / min. or exceeding 0, 2, 4, 6, 8, 10, 12, 14, 16 or 18 μL / min Good. In some cases, the flow rates in the second domain in standby mode are 20, 18, 1 The flow rate may be less than 6, 14, 12, 10, 8, 6, 4, or 2 μL / min.

[0115] During the dispensing mode, when droplets are being dispensed from the tip, the flow rate in the second region is 10-100 μF. The rate is L / min, or 10, 20, 30, 40, 50, 60, 70, 80 or 90 μL. It may exceed / min. In some cases, the flow rate in the second zone between dispensing may be 100, 90, 8 The flow rate is less than 0, 70, 60, 50, 40, 30, 20, or 15 μL / min.

[0116] The droplets may contain biological material, cells, or beads. The droplets may contain a single cell or multiple cells. The droplet may contain cells. The droplet may contain one or more beads. The droplet is optional. The shape or size can be such. However, preferably the droplet is spherical or It is cylindrical. The droplet size may be between 20 and 600 μm. However, The droplet sizes are 20, 30, 40, 50, 60, 80, 100, 120, 140, and 15. 0, 160, 180, 200, 220, 240, 250, 260, 280, 300, 32 0, 340, 360, 380, 400, 420, 440, 460, 480, 500, 52 It may exceed 0, 540, 550, 560, or 580 μm. In some embodiments... The droplet sizes are 600, 580, 560, 550, 540, 520, 500, and 4 80, 460, 440, 420, 400, 380, 360, 340, 320, 300, 2 80, 260, 250, 240, 220, 200, 180, 160, 150, 140, 1 The size may be 20, 100, 80, 60, 50, 40, or less than 30 μm. Multiple droplets They can be combined to form larger droplets. Alternatively, a large droplet can be divided. This allows for the formation of smaller droplets as needed.

[0117] If the droplet is too small relative to the height of the microfluidic chamber, then the droplet will be too small. The droplets are not in contact with the inner walls of the fluid chamber on either side. Therefore, the droplets are not in contact with the oEWOD. It cannot be moved. In contrast, a large droplet relative to the device's geometry is microscopic. Moving within the small fluid chamber may become difficult and / or slow. This can interfere with other droplets of the appropriate size, or cause them to merge with droplets of the correct size. This often simply gets in the way and disrupts other operations.

[0118] Referring to Figure 1, the second region 16 contains an inlet port and an outlet port within the second region 16. These are channels such as minute channels connected between the terminal and the terminal. The minute channel 16 is a minute channel. The droplet is received from the first region 14 through the gap 20 inside the constricting means 18. The microdroplets are then subjected to a higher carrier fluid flow rate in the second region 16 of the microfluidic chip 10. It is configured to move to the outlet port. This higher carrier fluid flow rate is in the second region. Generated by attaching the pump source to one of the multiple ports located inside 16 The pump source may be one or more inlet or outlet ports of the microfluidic tip 10. It may be a syringe pump or a high-pressure pump connected to the mouth port. In addition, the valve , software connected to one or more outlet or inlet ports of the microfluidic chip 10 It can be used as a tower control valve.

[0119] The microchannel is connected between the inlet port and outlet port in the second region 16. In the following manner, it is possible to pattern the inside of the second region 16 of the microfluidic chip 10. ru.

[0120] As shown in Figure 1, the pump source is connected to ports 22 and 28, etc., through a two-way valve. It is connected to one or more outlet ports. The droplets are drawn in from port 22. For example, it is loaded through another port, which is port 24. The droplet is in the first region 14 It is manipulated in and then selected, and moved to the second region 16 via the application of the oEWOD force. Figure 1 also shows that the droplet then exits the outlet port, which is, for example, the outlet port 26. The contents are dispensed by pumping into port 28, while ports 22 and 24 are dispensed through valves. This indicates that it will be shut off. Then the pump source is switched off, and the output of the microfluidic tip 10 The valve in the oral port is in the closed position.

[0121] Referring to Figures 2A, 2B, and 2C, microdroplets are loaded onto the microfluidic chip 10. 30, and the dispensing sequence are shown. The microfluidic tip 10 contains the enclosed volume 12. The enclosed volume 12 includes the first region 14 and the second region 16, as shown in Figure 2A. The valve 32 allows the microfluidic chip 10 to open and close each of the ports 22, 24, 26, and 28. It is connected to ports 22, 24, 26, and 28. The droplet 30 is connected to the carrier fluid containing the droplet. By passing the flow between these two ports, the first region 14 of the chip 10 It is loaded into the first region 14. Meanwhile, the other ports are sealed by valve 32. To reduce the flow rate to zero, valve 32 is closed. In this case, droplet 30 is in the first region. It is stored in region 14 and / or manipulated in the first region 14.

[0122] Referring to Figure 2B, droplet 30 is selected and, by the application of force, constricts the hand. The object is moved from the first region 14 to the second region 16 through the gap 20 located inside the step 18.

[0123] As shown in Figure 2C, the droplet 30 is then pumped to port 28 using the syringe pump 35. By pumping it internally, it is dispensed from the outlet port, for example, the outlet port 26. Meanwhile, ports 22 and 24 are shut off via valve 32, as shown in Figure 2C. To enable this, the droplet 30 is dispensed into a receptacle such as a multiwell plate 34. This is possible. The voltage is turned off during the dispensing cycle, and the droplet is released from the oEWOD force. It may be possible to make that possible.

[0124] Referring to Figure 3A, a microfluidic chip 10 showing the second region 16 is provided. Ports 26 and 28 of the chip are connected to valve 32. Both valves are open as shown in Figure 3A. It is in position. Carrier fluid is injected from pump 35, and droplets 30 pass through the second region 16. It moves and exits from tip 10 into conduit 40. The outlet valve 32 is connected to the waste channel 36 or container. It is open to the conduit. Sensor 38 monitors the presence of droplets inside the conduit 40. It is placed in the neighborhood of 40.

[0125] The detection module 38 is an optical sensor such as a photodiode or phototransistor. , or electrical sensors such as capacitive sensors or impedance sensors, or This can be a combination of several sensors. The set of sensors is located near the conduit. It can be placed around the sensor. In this case, the sensor detects when a droplet passes through the detection window. This will generate an electrical signal. Optionally, to image the inside of the tubes on both sides of the valve. An inspection camera is placed, and video or images from the inspection camera are read by a reader module. It can be recorded and analyzed.

[0126] The device further reads the generated signal from the sensor or detection module and controls A microcontroller configured to transmit to the roller (as shown in the attached drawing) It is equipped with a leader module such as a microcontroller. This reads the sensor's output signal and sends the sensor's status to the controller. It is composed.

[0127] Referring to Figure 3B, droplet detection using a sensor or optical detector 38 is illustrated. The software controller controls the receptacle so that the droplet moves into the receptacle 34. The valve 32 is positioned to open to the second conduit 42 directed into the tackle 34.

[0128] Referring to Figure 4A, the operation and assay are performed within the optical fluid chamber inside the first region 14. The target droplet 43 is shown. The droplet 43 can be moved and rearranged in the array. ru.

[0129] Referring to Figures 4B and 4C, the selected droplet 43 is narrowed by the oEWOD force. It is shown that the object is moving to the second region 16 through the narrowing means 18.

[0130] As shown in Figure 4D, the valve connected to the port of the second region 16 is opened, and the carrier flow The body is injected to generate a high flow rate inside the second region 16, and the droplets are carried out of the device. Therefore, droplet 43 is dispensed.

[0131] Referring to Figure 5A, droplets can be dispensed in a multiphase flow. One is an aqueous medium 44. The two independent pumps, one of which has an immiscible carrier fluid 46, are probably The valve 3 is connected to the inlet port 48 by a merging component 47. As shown in Figure 5A, valve 3 2 is also provided. The valves 32 can be opened and closed independently of each other. In some cases, the valves are in sequence. They may open and close in tandem, or they may open and close simultaneously. The lug is placed before and / or after the volume of the immiscible carrier fluid in the second region 1 of the tip 10. Inject into 6. Move the droplet 30 to the immiscible carrier fluid portion. Then, the mixed phase flow The body is injected into the dispensing receptacle through outlet 49. This allows the dispensing receptacle to The volume of the immiscible fluid introduced into the system decreases.

[0132] As shown in Figure 5B, smaller droplets 54 are selected and combined to form larger droplets. By forming droplets 52, larger droplets 52 can be created. The combined droplets 52 may have a diameter of approximately 50 μm. In some cases, When a voltage between 5 and 10V, preferably 10V, is applied to the device, larger droplets 52 are formed. Smaller droplets 54 can be combined to achieve this. The combined droplets 52 By using a pump, the microfluidic fluid can be pushed out from the outlet port of the microfluidic tip. The flow rate within the second region may be between 10 and 100 μL / min.

[0133] In some cases, if the droplets are mixed with the water stream (or plug) immediately after leaving the tip, The amount of oil that must ultimately be discharged from the tip and into the well, This minimizes the amount of oil that can be absorbed into receptacles such as wells. This would help avoid this. Additionally or alternatively, small droplets 54 can be used to separate large aqueous particles. Lug 52 or combined droplets can precede the microfluidic fluid via the syringe pump. It can be pumped out from the tip. Therefore, the receptacle such as a well can be filled with oil. The condition is avoided from being met.

[0134] Referring to Figure 6, a dispensing device or system 100 is provided. Tem 100 comprises the devices disclosed in the aforementioned embodiments of the present invention. One or more The device 100, which dispenses several tiny droplets, is equipped with a microfluidic tip 102. 102(A) includes a first region and a second region. The first region and the second region are constricting means It is separated by the following. The first region is one or more microliquids dispersed in the carrier fluid. The droplet is adapted to receive and manipulate at a low carrier fluid flow rate. The second region is narrowed. The system receives microdroplets from the first region via a step, and the microdroplets are then transported to a higher carrier fluid flow rate. The second region is configured to move to the exit port of the microfluidic chip. By applying an electrowetting (oEWOD) force through the constriction means, It is configured to receive the microdroplet from the first region.

[0135] The device further includes a controller. The controller controls the outlet port of the microfluidic chip 102. Configured to control valves and / or pumps connected to the t, as shown in Figure 6. Valve 103(B) is connected to the outlet port of the microfluidic device 102. (See attached drawing) The pump (not shown) is connected to the inlet port of the microfluidic device 102. The outlet port of the fluid tip 102 is connected to a conduit 104, such as a tube. The conduit is transparent. It can be done this way.

[0136] Receptacle 108 is a multiwell plate 108. A multiwell plate is It is mounted on a multi-axis controlled stage 110 in an XYZ configuration. Multiwell plate 108 This may be a 96 or 384-well plate. Stage 110 can be operated manually or automatically. It can be precisely controlled. Receptacle 108 is a waste container, reservoir, PCR tube. It can also be a microcentrifuge tube such as a tubing or Eppendorf tubing. Optional For selection, each well was pre-filled with a certain amount of cell culture medium. The controller was used during the dispensing procedure. The system is configured to control the movement of the stage on which the multiwell plate is placed. Dispensing one droplet into a well, and / or dispensing multiple droplets into a single well. It is possible.

[0137] During the dispensing procedure, the dispensing head 106 descends into the well containing aqueous buffer. Alternatively, the well may move toward the dispensing head 106. Fix the head 106 in place and move the well plate toward the dispensing head 106. It may be done. The pump connected to the inlet port of the microfluidic chip 102 is operated, and micro To pump the required amount of buffer through the fluid tip, a considerable amount of time is needed, at an appropriate speed. Pump in degrees. The exact time and speed depend on the size of the microchannels, the interconnecting tubes, and it varies depending on the interface connection part. For example, the inlet port of the microfluidic chip 102 The pump connected to the terminal can be activated and pumped at 50 μL / min for 12 seconds. The valve connected to the outlet port of the microfluidic tip 102 typically handles approximately 7 μL to 10 μL. A certain volume L is pushed from the microfluidic tip into tube 104 and into the waste container. It is opened to allow dispensing. A fixed volume of 7 μL to 10 μL is provided by a microchannel. and sufficient to completely purge the interconnecting tubes and interface connections. That's good too.

[0138] Next, one or more droplets are released from the microfluidic tip 102, stopping just before the valve. It can be moved into tube 104. Then the pump is deactivated, and the microfluidic fluid is released. The pumping of fluid from device 102 is stopped. Meanwhile, the valve moves to the dispensing stop position. The pump is restarted by the controller for another 4 seconds. The droplets are in well 108. It is dispensed. Then the valve is closed manually or by a software-controlled controller. The valve can be closed automatically.

[0139] In some examples, the method or sequence for dispensing droplets is as follows: This can be done by switching off the pump source. The valve is controlled by the controller. It is in a closed position so that it is controlled. It manipulates the target droplet and the optical flow inside the microfluidic chip. The assay is performed in a body chamber. Optoelectrowetting transfer is used to transfer the target droplet to light. The fluid is moved from the fluid chamber into the microchannels inside the microfluidic chip. The 3-axis stage is , multiwell so that the target well is located below the outlet tube of the software-controlled valve Move the plate. Activate the software-controlled pump, typically at 7 micrometers. Start the replacement of a fixed volume of fluid in liters, microchannels and interconnecting tubes. And properly purge the interface connection.

[0140] Next, a software-controlled valve is used to route the fluid into the multiwell plate. Switch to the open position. The resulting fluid flow in the microchannels moves the droplets. The carrier phase volume is purged into the multiwell plate. Optionally, sensors or The camera is consulted, and before the multiwell plate is determined, the outlet tube... Check for the presence of droplets. If no droplets are detected, the pump source will be used to collect the droplets. Command to dispense the correct amount. Switch off the pump and close the valve. Dispensing head Remove from the multiwell plate and / or put the multiwell plate into the dispensing head. Pull it out from the dispenser. Optionally, move the multiwell plate and place it under the dispensing head. Place a waste well or alternative waste receptacle to purge the microfluidic pathway. Repeat the above steps until all target droplets are recovered from the microfluidic device. The well plates were collected for further experiments such as DNA sequencing or cell expansion. ru.

[0141] Alternatively, by relying on pumps and valves to measure the correct amount and collect the droplets. The droplets can be collected. As just one example, a 20cm tube length, 0.1mm To provide a measuring volume of 2 to 5 μL for dispensing 0.1 μm at an inner diameter of 20 μl / min. This can be done. This requires providing a sensor or camera to detect droplets inside the tube. This means that there is no additional or alternative device disclosed in this invention. Alternatively, to parallelize with the collection of multiple droplets, multiple dispensing routes and multiple points may be used as appropriate. It can support the pump source and valve.

[0142] The devices, apparatus, and methods of the present invention are used in many applications, such as single-cell dispensing. It is possible. In some cases, the droplet may contain multiple cells. The droplet may contain a single cell. It may contain a random number of cells including [specific cells]. Furthermore, it may contain a single cell or multiple cells. The recovered droplets were subjected to PCR amplification, DNA sequencing, RNA sequencing, and cell expansion. Assays can be performed that may include, but are not limited to, large amounts. The efficiency of dispensing is, liquid The droplets are stained with trypan blue, and the droplets are filmed using a camera before and after the dispensing valve. This can be evaluated by the following: For example, if the dispensing valve is used less than 12 seconds after dispensing begins Dispensing is considered successful if droplets are detected later. In only one example. The efficiency of performing PCR by dispensing single cells is approximately 80% (40 / 50), while dispensing The overall efficiency of subsequent PCR tests was 79% (66 / 84).

[0143] Various further aspects and embodiments of the present invention will be apparent to those skilled in the art from the viewpoint of this disclosure. be.

[0144] "and / or" as used herein refers to two specified features or configurations. Each element should be considered a specific disclosure. For example, “A and / or B” (i) A, (ii) B, and (iii) A and B, each as if each of these items It should be considered a specific disclosure, as if it were individually described in the document.

[0145] Unless otherwise indicated in the context, the above descriptions and definitions of features apply to specific embodiments of the present invention. This applies equally to all aspects and embodiments described, and is not limited to those embodiments. .

[0146] Furthermore, although the present invention has been described by example with reference to several embodiments, the disclosure Not limited to the embodiments described herein, and not departing from the scope of the invention as defined in the appended claims. Those skilled in the art will understand that alternative embodiments can be constructed without this.

Claims

1. The system is configured to generate optically mediated electrowetting (oEWOD) forces. A microfluidic chip having a constructed oEWOD structure comprises one or more microdroplets A dispensing device, wherein the microfluidic chip includes a first region and a second region, The first and second regions are separated by a constriction. The first region receives one or more microdroplets dispersed in the carrier fluid at a first flow rate. Adapted to be taken and operated, The second region receives minute droplets from the first region through the constricted portion, and the minute droplets are then... It is configured to move the microfluid to the outlet port of the microfluidic tip at a flow rate of 2. The second region is the application of optically mediated electrowetting (oEWOD) forces. Therefore, it is configured to receive the minute droplets from the first region through the constricted portion, The device includes a controller, and the controller controls the second region of the second region. The flow rate is set to be higher than the first flow rate in the first region by using valves and / or pumps. A device configured to control something.

2. A device according to claim 1, wherein the constricted portion is a physical barrier.

3. A device according to claim 1, wherein the constricted portion is a semipermeable membrane.

4. A device according to any one of claims 1 to 3, wherein the microdroplets are not a single biomaterial. A device containing multiple cells, or one or more beads.

5. The device according to claim 1 or 2, wherein the constricted portion is provided with an opening, and the opening The device has a width between 20 and 400 microns.

6. A device according to any one of claims 1 to 5, wherein the geometry of the second region is substantially A device with a crescent-shaped channel.

7. A device according to any one of claims 1 to 6, wherein the second region comprises a plurality of channels Furthermore, each channel receives a microdroplet from the first region and flows the microdroplet through the microstream. A device configured to move to the exit port of the body chip.

8. A device according to any one of claims 1 to 7, wherein the valve and / or pump is a guide A device configured to be connected by a tube to the outlet port of the microfluidic chip. vinegar.

9. The device according to claim 8, wherein the outlet port of the microfluidic chip is connected The system further comprises a controller configured to control the valve and / or pump, device.

10. The device according to claim 9, wherein the controller has a channel in the second region. In each of these, the flow of the microdroplets, or the flow of each individual microdroplet, is controlled. A device composed of the following.

11. A device according to any one of claims 1 to 10, wherein a plurality of microdroplets are the micro A device that is simultaneously moved to the outlet port of the fluid chip.

12. A device according to any one of claims 1 to 11, wherein the inlet port of the first region or The system further comprises an outlet port and a valve provided at the inlet port or outlet port of the first region. ,device.

13. A device according to any one of claims 1 to 12, wherein the detection signal is transmitted to the microfluidic chip The system further includes a detection system that detects from minute droplets dispensed from the outlet port of the bottle. device.

14. The device according to claim 9, wherein the signal generated from the sensor or detection module The system further comprises a reader module configured to read and transmit data to the controller. Furthermore, the controller positions the valve in the open position so that minute droplets are dispensed. A device that is further configured in this way.

15. A device according to any one of claims 1 to 14, further comprising a receptacle, The receptacle is a device configured to receive dispensed microdroplets.

16. The device according to claim 15, wherein the receptacle is a multiwell plate, A device that is a PCR tube or microcentrifuge tube.

17. The device according to claim 16, wherein the multiwell plate is multi-axis motion control type The multi-axis motion-controlled stage is mounted on a stage, and the target well is the microfluidic chip The multiwell plate is positioned below the valve provided at the outlet port of the tubing. A device configured to move to a first position.

18. The device according to claim 13, wherein the detection system includes an optical detector. 。

19. The device according to claim 17, wherein each well is pre-filled with a certain amount of cell culture medium. A device.

20. The device according to claim 19, wherein each well is pre-filled with a certain amount of buffer, water, or oil. A device that is filled with liquid.

21. A device according to claim 1, wherein the constricted portion is a sheath fluid.

22. A method for dispensing one or more microdroplets, A microfluidic chip is provided, comprising a first region and a second region separated by a constricted portion. The steps, The process includes the step of transferring a microdroplet from a first region to a second region, wherein the microdroplet is in the first region In the first region, the first region is dispersed in the carrier fluid at a first flow rate, and the second region is separated from the first region by a constricting means. It receives microdroplets from and carries the microfluidic chips at a higher carrier fluid flow rate. It is configured to move to the exit port of the P. The second region is the application of optically mediated electrowetting (oEWOD) forces. Therefore, it is configured to receive the minute droplets from the first region through the constricted portion, The second flow rate in the second region is set to be higher than the first flow rate in the first region. To control the flow of the carrier fluid, a controller is used with valves and / or pumps. A method including the steps of activating the device.

23. A method according to claim 22, wherein the controller is used to control the microfluidic chip To control the flow of carrier fluid through the outlet port, a pump and / or valve are activated. A method that includes further steps to achieve this.

24. The method according to claims 22 and 23, wherein the multiwell plate is a multi-axis motion control type A step placed on the stage, wherein the multi-axis motion-controlled stage is the controller The multi-well plate is configured to be moved to the target well using a ra, The step where the target well is located below the valve provided at the outlet port of the microfluidic tip valve Furthermore, methods.

25. The method according to claim 24, wherein microdroplets are dispensed onto the multiwell plate. A method further comprising the step of switching the valve to the open position.

26. A method according to claim 24, wherein the target well is recorded using the controller. A method that includes further steps.

27. A method according to any one of claims 22 to 26, wherein the detection module or sensor A method further comprising the step of generating a signal using

28. A method according to claim 27, wherein the generated signal is transmitted to the detection module or the front The steps include detecting the signal from the sensor and transmitting the generated signal to the controller. The step, and further including, the controller, valve so that minute droplets are dispensed. A method further configured to switch to the open position.

29. A device for dispensing one or more microdroplets, A microfluidic chip according to claim 1, wherein microdroplets dispersed in a carrier fluid, A microfluid including a second region configured to be transferred to the outlet port of the microfluidic chip. Tips and, The microfluid passes through the microfluid chip from the inlet port to the outlet port. A pump configured to control the flow of the carrier fluid, A conduit connected to the outlet port of the microfluidic chip, once the chip A conduit that receives the microdroplets dispensed from, A sensor positioned near the conduit configured to generate a signal, The system is configured to read the generated signal from the sensor and transmit it to the controller. Equipped with a leader module, The controller has a valve and / or a valve connected to the outlet port of the microfluidic chip. or is configured to control the pump, In response to the signal generated by the sensor, the controller determines that the minute droplet is The device is configured to switch the valve to the position where the contents are dispensed, or the controller The valve is configured to switch to a position where a microdroplet is dispensed onto the receptacle. A device.

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