CN102281950A - Programmable microfluidic digital array - Google Patents

Programmable microfluidic digital array Download PDF

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Publication number
CN102281950A
CN102281950A CN200980154761XA CN200980154761A CN102281950A CN 102281950 A CN102281950 A CN 102281950A CN 200980154761X A CN200980154761X A CN 200980154761XA CN 200980154761 A CN200980154761 A CN 200980154761A CN 102281950 A CN102281950 A CN 102281950A
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valve
fluid
input
microfluidic device
group
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CN200980154761XA
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CN102281950B (en
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布赖恩·福勒
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Standard Biotools Corp
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Fluidigm Corp
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Priority to CN201510158081.2A priority Critical patent/CN104741159B/en
Priority to CN201610459013.4A priority patent/CN105964316A/en
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
    • B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L7/00—Heating or cooling apparatus; Heat insulating devices
    • B01L7/52—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples
    • B01L7/525—Heating or cooling apparatus; Heat insulating devices with provision for submitting samples to a predetermined sequence of different temperatures, e.g. for treating nucleic acid samples with physical movement of samples between temperature zones
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00281—Individual reactor vessels
    • B01J2219/00286—Reactor vessels with top and bottom openings
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00279—Features relating to reactor vessels
    • B01J2219/00306—Reactor vessels in a multiple arrangement
    • B01J2219/00313—Reactor vessels in a multiple arrangement the reactor vessels being formed by arrays of wells in blocks
    • B01J2219/00315—Microtiter plates
    • B01J2219/00317—Microwell devices, i.e. having large numbers of wells
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00389—Feeding through valves
    • B01J2219/00396—Membrane valves
    • B01J2219/00398—Membrane valves in multiple arrangements
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00418—Means for dispensing and evacuation of reagents using pressure
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/02—Adapting objects or devices to another
    • B01L2200/026—Fluid interfacing between devices or objects, e.g. connectors, inlet details
    • B01L2200/027—Fluid interfacing between devices or objects, e.g. connectors, inlet details for microfluidic devices
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0816—Cards, e.g. flat sample carriers usually with flow in two horizontal directions
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/089—Virtual walls for guiding liquids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/18—Means for temperature control
    • B01L2300/1805—Conductive heating, heat from thermostatted solids is conducted to receptacles, e.g. heating plates, blocks
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0415—Moving fluids with specific forces or mechanical means specific forces electrical forces, e.g. electrokinetic
    • B01L2400/0427—Electrowetting

Abstract

A microfluidic device includes a pressure source and a control line in fluid communication with the pressure source. The microfluidic device also includes a plurality of valves operated via the control line and an independent valve positioned adjacent the control line and between the pressure source and the plurality of valves.

Description

The programmable microfluidic digital array
Background of invention
Be used on the yardstick that microfluidic device can still can't be imagined before soon analyzing, prepare, measure and other operating functions.The advantage of microfluidic device comprises saves the fluid accuracy and the degree of accuracy on precious reagent and sample, sample analysis or synthetic high density and high flux, the visible hardly level of naked eyes and replaces the space that corresponding device brought of operating and reduce on the macrofluid yardstick.Reducing to increase relevant with density with the microfluidic device size is higher complexity and bigger engineering and manufacturing cost and meticulousr device construction.
Although have these to make progress in microfluid design and application facet, the operation that reduces the complexity of micro-fluid chip and simplify them still is useful.In addition, the control fluid that need to strengthen flow and microfluidic device in the ability of the correlated response process that takes place.Therefore, the improved method and system that this area need be relevant with microfluidic device.
Summary of the invention
The present invention relates to microfluidic device.More specifically, the present invention relates to programmable microfluidic digital array and method of operating thereof.Only as an example, described method and apparatus has been applied to provide in micro-fluid chip in the system of asynchronous logic function.In addition, embodiments more of the present invention are integrated into digital array so that the locking control line to be provided with check valve.Yet, will be appreciated that the present invention has the much wide scope of application.
According to one embodiment of the invention, provide microfluidic device.The control line that described microfluidic device comprises pressure source and is communicated with the pressure source fluid.Described microfluidic device also comprises a plurality of valves by described control line operation, also comprises separate valves, and described separate valves is near the described control line and between described pressure source and described a plurality of valve.
According to another embodiment of the invention, provide operation to have the method for the microfluidic device of valve and control line, described control line has coupled valve group.Described method comprises closes described valve and described control line is exerted pressure.The valve group that described valve of cutting out causes linking to each other with described control line can not be operated.
According to another embodiment of the present invention, provide microfluidic device.The control line that described microfluidic device comprises first valve, second valve and is communicated with described first valve and the second valve fluid.Described microfluidic device also comprises acute build up of pressure device (pressure accumulator) and near the check valve the described control line between described acute build up of pressure device and described second valve that is communicated with described control line fluid.
According to an alternate embodiment of the present invention, provide microfluidic device.Described microfluidic device comprises a plurality of reative cells of arranging with array layout.Each of described a plurality of reative cells have with a plurality of reative cells in one of first valve that is communicated with of fluid and second valve that is communicated with one of described fluid in described a plurality of reative cells.Described microfluidic device also comprises can be operated with first control line that starts described first valve and described second valve and the input line group that is communicated with described a plurality of reative cell fluids.Described microfluidic device also comprises a plurality of sample inlets that are communicated with described input line group fluid and is arranged in check valve in described first control line.
According to another alternate embodiment of the present invention, provide microfluidic device.Described microfluidic device comprises a plurality of reative cells and a plurality of first input port.Each of described a plurality of first input ports is communicated with one or more fluids of described a plurality of reative cells by one of more than first input lines.Described microfluidic device also comprises a plurality of second input ports.Each of described a plurality of second input ports is communicated with one or more fluids of described a plurality of reative cells by one of more than second input lines.Described microfluidic device also comprises the first acute build up of pressure device that is communicated with the first control line fluid.Described first control line is set to close described more than first input lines.In addition, described microfluidic device comprises the second acute build up of pressure device that is communicated with the second control line fluid.Described second control line is set to close described more than second input lines.In addition, described microfluidic device comprises first check valve in described first control line that is arranged between described first acute build up of pressure device and described more than the first individual input lines, and is arranged in second check valve in described second acute build up of pressure device and described more than second individual described second control lines of importing between the lines.
According to another alternate embodiment, provide operation to have the method for the microfluidic device of a plurality of valves and check valve.Described method comprises that the control line to described microfluidic device applies first fluid pressure, and responds to described first the applying of pressure and close described a plurality of valve.Described method also comprises and responds to described first the applying of pressure and close described check valve, and applies second fluid pressure to second control line of described microfluidic device.
According to a specific embodiments of the present invention, provide operation to have the method for the microfluidic device of a plurality of input ports.Described method comprises to one of described a plurality of input ports provides the input fluid, and starts the valve group to close the first's input line that is connected with the subgroup of described a plurality of input ports.Described subgroup does not comprise the one of described of described a plurality of input ports.Described method also comprises makes described input fluid flow through one of the described input line that is connected with described a plurality of input ports, and makes the described input fluid described input line of flowing through arrive the second portion of described input line and close the second valve group to separate a plurality of reative cells.
According to another specific embodiments of the present invention, the method for operation programmable microfluidic device is provided, described device has the reaction site array that is communicated with the first input line group and the second input line group fluid.Described method comprises starting can operate in order to blocking the first valve group of the described first input line group, and starts the second valve group that can operate in order to the first of the subgroup of blocking the second input line group.Described method comprises that also the second portion by the described second input line group is loaded into described reaction site with sample and starts and can operate in order to separate the 3rd valve group of described reaction site.
According to another specific embodiments of the present invention, provide microfluidic device.Described microfluidic device comprises input port and a plurality of input fluid line of predetermined quantity, and each of described input port can be operated to receive one of a plurality of input fluids, and each of described a plurality of input fluid lines is communicated with one of the input port of described predetermined quantity fluid.Described microfluidic device also comprises the valve group, and each of described valve group can be operated to close one of described a plurality of input fluid lines.The quantity of described valve group is less than described predetermined quantity.Described microfluidic device also comprises the manifold and the second valve group that is communicated with each fluid of described input fluid line, and each of the described second valve group can be operated to close the part of described manifold.
According to one embodiment of the invention, the method for operation programmable microfluidic device is provided, described device has the reaction site array, and it is communicated with the first input line group, the second input line group and the manifold fluid that is connected with the described second input line group.Described method comprises starting can operate in order to close the first valve group of the described first input line group, start the second valve group that to operate in order to the first of the subgroup of closing the second input line group, and start and to be operated with the 3rd valve group of (deactivate) so that described manifold is stopped using.Described method also comprises cancelling starts (deactuate) described second valve group, and the second portion of a plurality of samples by the described second input line group is loaded into described reaction site, and starts and can be operated in order to separate the 4th valve group of described reaction site.
According to another embodiment of the invention, the method for operation programmable microfluidic device is provided, described device has the reaction site array that is communicated with the first input line group and the second input line group fluid.Described method comprises starting and can be operated in order to blocking the first valve group of the described first input line group, and starts and can be operated in order to separate the second valve group of described reaction site.Described method also comprises cancelling starts the described second valve group, and a plurality of samples are written into described reaction site and start the described second valve group by the second input line assembling.
According to another embodiment of the present invention, provide microfluidic device.Described microfluidic device comprises a plurality of reaction site and the first input line group, and the described first input line group provides first input port of predetermined quantity and the fluid between described a plurality of reaction site to be communicated with.Described first group quantity is described predetermined quantity.Described microfluidic device also comprises the second input line group, and it provides second input port of predetermined quantity to be communicated with fluid between described a plurality of reative cells.Each of the described second input line group comprises trunk portion and component, and described second group quantity is less than described predetermined quantity.Described microfluidic device also comprises programmable input unit, and it can be operated in order to use described first input line group or the described second input line assembling to fill out described reative cell.
According to a specific embodiments of the present invention, provide the method that the microfluidic device with a plurality of control lines is set.Described method comprises starts first control line and valve is placed first state.Start after described method also comprises and to be operated in order to the valve group is placed second control line of second state.The valve that is in described first state stops described valve group to be placed in second state.
According to another specific embodiments of the present invention, provide the method that the microfluidic device with a plurality of control lines is set.Described method comprises by starting first control line and start first state that second control line is set up described microfluidic device then, and by starting described second control line and starting second state that described first control line is set up described microfluidic device subsequently.
According to another specific embodiments of the present invention, provide a kind of microfluidic device.Described microfluidic device comprises first valve and second valve.Described microfluidic device also comprises the control line that is communicated with described first valve and the described second valve fluid, and the pressure source that is communicated with described control line fluid.Described microfluidic device also comprises the check valve that is arranged between pressure source described in the described control line and described second valve.
According to another alternate embodiment of the present invention, provide microfluid system.Described microfluid system comprises carrier.Described carrier comprises a plurality of first input ports and a plurality of first input line.Each of described a plurality of first input lines is communicated with one of described a plurality of first input ports fluid.Described carrier also comprises a plurality of second input ports and a plurality of second input line.Each of described a plurality of second input lines is communicated with one of described a plurality of second input ports fluid.Described carrier also comprises first pressure source and second pressure source.Described microfluid system also comprises the microfluidic device that is contained on the described carrier.Described microfluidic device comprises a plurality of the 3rd input lines and a plurality of the 4th input line.Each of described a plurality of the 3rd input lines is communicated with one of described a plurality of first input lines fluid, and each of described the 4th input line is communicated with one of described a plurality of second input lines fluid.Described microfluidic device also comprises first control line that is communicated with the described first pressure source fluid, can be operated in order to check valve that blocks described first control line at least a portion and second control line that is communicated with the described second pressure source fluid.
The present invention has the many advantages that surmount routine techniques.For example, the technology of the present invention allows to customize microfluidic device after manufacturing, makes the user that the setting of a plurality of panels able to programme can be provided.These and other embodiments of the present invention with and the contact of many advantages and feature hereinafter carried out more detailed description with accompanying drawing.
The accompanying drawing summary
Fig. 1 is the rough schematic view of microfluidic device according to an embodiment of the invention.
Fig. 2 A is the rough schematic view of check valve according to an embodiment of the invention.
Fig. 2 B is the simplification top view that shows check valve shown in Fig. 2 A.
Fig. 3 is the simplified flow chart of the method for a kind of operation microfluidic device according to an embodiment of the invention.
Fig. 4 is the simplified flow chart of the method for a kind of operation microfluidic device according to another embodiment of the invention.
Fig. 5 is the simplified flow chart of method of the microfluidic device of a kind of operation another embodiment according to the present invention.
Fig. 6 A is the sketch of reative cell in according to an embodiment of the invention the array.
Fig. 6 B is the simplified perspective view of control line in according to an embodiment of the invention the array, fluid input line and reative cell.
Fig. 7 shows a kind of method for simplifying that programmable microfluidic device is according to an embodiment of the invention programmed.
Fig. 8 shows a kind of method for simplifying that programmable microfluidic device is according to another embodiment of the invention programmed.
Fig. 9 shows a kind of method for simplifying that the programmable microfluidic device of another embodiment according to the present invention is programmed.
Figure 10 is the rough schematic view of programmable microfluidic device according to an embodiment of the invention.
Figure 11 is the rough schematic view of microfluid system according to an embodiment of the invention.
Specific embodiments describes in detail
Fig. 1 is the rough schematic view of microfluidic device according to an embodiment of the invention.In a specific embodiment, microfluidic device shown in Figure 1 comprises high-density digital array able to programme.Described microfluidic device 100 comprises first pressure source 102 and second pressure source 104.In Fig. 1, first pressure source 102 is called as and holds accumulator (containment accumulator), as hereinafter described in more detail, described pressure source 102 is communicated with the control line fluid, and described control line can be operated the valve that is associated with reative cell (not shown) in the array 106 in order to close.Because starting that carry out in working pressure source 102 makes these reative cell valves close and make sample and/or reagent to be contained in the described reative cell, therefore pressure source 102 is called and holds accumulator in some embodiments.
In operation, liquid is placed in acute build up of pressure device (pressure accumulator) or the pressure source, described acute build up of pressure device or pressure source link to each other with outside malleation supply (as containing compressed-air actuated container) subsequently.Under pressure, described compressed air or other fluids push control line with the liquid in the described acute build up of pressure device, thereby start valve.Therefore, described pressure source does not comprise pressure fluid during fabrication usually, but the container of control line being exerted pressure with after activation can be accumulated and store to pressure fluid therein during being provided at the operation microfluidic device.As more complete description in the application's full text, described acute build up of pressure device can be operated to keep the pressure in the control line after activation.At common unsettled and common transfer U.S. Provisional Patent Application No.61/044, the additional description of the digital array of suitable enforcement embodiment of the present invention is provided in 417, its whole disclosures are quoted by integral body all purposes and are incorporated this paper into.
Fig. 6 A is the sketch of reative cell in according to an embodiment of the invention the array.As an example, embodiments more of the present invention adopt the cell with reative cell, and its lateral dimension is 100 μ m * 60 μ m, highly is 125 μ m.In this exemplary embodiment, the volume of described chamber is about 0.75nl.So little chamber volume energy uses littler sample volume and has reduced running cost.Through hole (via) the coupled reaction chamber of diameter 50 μ m and mensuration/sample input line.With the first side direction chamber spacing of 110 μ m and the spacing placing response chamber, the second side direction chamber of 200 μ m.The valve of 50 μ m * 50 μ m is provided in the layer different with the layer that contains mensuration/sample input line, and described valve can be operated in order to the prevention described input line of flowing through.The particular geometric profile of this cell is not intended to limit embodiment of the present invention, and only provides the example of specific embodiments.In another embodiment, adopt other device geometric shapes that are suitable for concrete application.
According to embodiments more of the present invention, adopt width range that the fluid of the described microfluidic device of flowing through is provided to the input line of about 75 μ m for about 5 μ m to about 400 μ m, depth bounds for about 5 μ m.Width range is about 5 μ m, and to be about 5 μ m be used for closing (valve off) by valve to the control line of about 75 μ m flows through the fluid of input line to about 400 μ m, depth bounds.Adopting width range in some embodiments is that about 10 μ m are that extremely about 500 μ m, altitude range are about the reative cell of 5 μ m to about 500 μ m to about 10 μ m to about 500 μ m, length.The geometric shape of these devices provides by the mode of example, and is not intended to limit embodiment as herein described.
As shown in Figure 6A, provide a plurality of input lines 620, made the fluid described input line of flowing through in the horizontal direction.Shown three input lines, but embodiments more of the present invention adopt the input line more than three, for example, 11 input lines.Adopt many input lines then single sample can be distributed in a plurality of input lines, this provides a plurality of copies of given sample.Described in the more complete description that Fig. 6 B is carried out, described input line or input channel to small part are included in the ground floor of microfluidic device.Referring to Fig. 1, described input line is communicated with mensuration input line 132 fluids in array 106 left sides, and is communicated with sample input line 142 fluids on array 106 right sides.Therefore, deriving from the fluid of measuring input 130 or sample input 140 all can offer the input line and and then offer reative cell.
A plurality of control lines 610 are provided in vertical direction, and the fluid that makes it possible to control through the input line flows.Shown two input lines, but embodiments more of the present invention adopt the control line more than two, for example, 70 control lines.In another embodiment, 71 control lines are arranged.Described many control lines form the reative cell that separates along the length of input line, and a plurality of reative cells that comprise same sample are provided.As to described in the more complete description of Fig. 6 B, described control line or control channel to small part is included in the second layer of microfluidic device.Referring to Fig. 1, described control line is communicated with 114 segment fluid flows of locking control line 110, described locking control line with hold accumulator 102 fluids and be communicated with.Form valve 615 at the intersection point of described control line and described input line, it responds to the fluid pressure in the control line and is started, and can operate to stop the fluid described input line of flowing through.Usually, but described multilayer microfluidic device comprises a plurality of elastomer layers and valve 615 comprises moveable diaphragm.In Fig. 6 A and 6B embodiments shown, but the moveable diaphragm of described valve deflects to the fluid passage on the intersection point that is arranged in control channel.Therefore, shown embodiment adopts " on push away " valve, but wherein moveable diaphragm upward deflects to the fluid passage to close the fluid passage in the valve position.For the valve that shows in Fig. 6 A and 6B, but thereby discharging the fluid pressure that exists in the control channel causes moveable diaphragm to return not inflection point opening the valve of closing.
Flow through the fluid of input line 620 along the direction vertical with Fig. 6 A plane through through hole 625 and upwards flow into reative cell 630, as in reative cell 630 to small part described in the more complete description of Fig. 6 B being included in the 3rd layer of microfluidic device.Therefore, described through hole to small part is included in described at least second or the 3rd layer of microfluidic device.Usually, adopt laser ablation to handle a part of removing second or the 3rd layer and form described through hole.Because described microfluidic device is that air is permeable, can adopts sealing filling technique (blind fill technique) filling reative cell and carry out number of chemical, biology or other experiments.Those skilled in the art understand, in reative cell, have fluid after, the starting of control line will cause valve to be closed and fluid will be kept the preset time section in reative cell.
Fig. 6 B is the simplified perspective view of control line, fluid input line and reative cell in the array of one embodiment of the invention.Array (for example, shown in Figure 1 array 106) is the part of multilayer microfluidic device.Every layer comprises the elastomer structure with one or more groove, passage, chamber etc. usually.As shown here, ground floor 601 comprises a plurality of control channels 610 that are arranged to array of parallel channels and other control channel 611.Described control channel 610 and other control channel 611 are communicated with one or more pressure source or acute build up of pressure device fluid.Therefore in one embodiment, control channel 610 is communicated with acute build up of pressure device 102 fluids, and control channel 611 is being communicated with acute build up of pressure device 104 fluids.Although shown single control channel 611 in Fig. 6 B, those of ordinary skills should understand shown single passage and represent one or more control channel that is associated with 601 layers.
In addition, described control channel is not restricted to and layer 601 relevant position.Described control channel can be adapted to concrete the application and place other layers.For example, in the enforcement of a multiple control (control-on-control), in order to make first control line (for example control channel 611) produce control, described second control line is placed on ground floor 601 and the second layer 602 among both according to the length of control channel to second control line (for example control channel 610).Adopt the through hole similar that second control line is transformed into the second layer 602 from ground floor 601 to through hole 625.By above first control line, passing second control line, at the intersection point formation valve of two control lines.In case start described first control line, the flexible membrane of valve position upwards promotes to be arranged in 602 layers second control line to block at valve position between then described first control line and described second control line.Therefore, as more complete description hereinafter, " on push away " valve can formed between the different control lines or between control line and the fluid input line.Referring to Fig. 1, the control line that links to each other with valve 128 carries out the transition to the upper strata by through hole from the lower floor of structure, makes control line 122 can start separator valve 124 thereby then pass control line 122.Then, thus the control line that is associated with valve 128 through another through hole wear get back in the ground floor by and start the valve of fluid input line 132.In case control line 122 is started, separator valve 124 blocks through the flowing of the control line that links to each other with valve 128, and prevention valve 128 responds to cutting out that control line 110 starts usually.
The second layer 602 comprises a plurality of fluid input channels 620 of also arranging with array of parallel channels.In the shown embodiment of Fig. 6 A and 6B, control channel 610 and input channel 620 are arranged with being perpendicular to one another.Exert pressure by the liquid (being generally basic incompressible fluid) that in control channel 610, exists and to realize starting of the valve that exists in the microfluidic device.Usually, liquid is placed in accumulator (accumulator) or the pressure source and the fluid (for example air, nitrogen etc.) of pressurization is provided to accumulator.The increase of pressure makes liquid enter control line under pressure in the accumulator.Respond to institute's applied pressure, but the moveable diaphragm that forms 601 layers of top upward deflects and enters fluid input channel 620.Therefore, produce " on push away " valve at control channel and fluid input channel intersection point.Some other embodiments can adopt " pushing away down ", " drop-down " or " on draw " valve by reorientating various control and fluid layer.
In embodiments more of the present invention of multiple control are provided, can provide one or more other key-course (for example lower floor 601), perhaps by control line is arranged into 602 layers.Described one or more other key-course can comprise other control line (not shown), and in a single day it start the control line that exists in the layer 601 closing by valve.Therefore, by the control fluid (for example liquid) that exists in one or more other control line is applied fluid pressure, flexible membrane deflects is to control line 610, stops the fluid control line of flowing through.Therefore, embodiments more of the present invention provide key-course, are also referred to as multiple control.Those of ordinary skills can understand many variations, improvement and replacement.
The described second layer also comprises a plurality of through holes 625, and producing provides the fluid passage of flowing from the fluid of three layer 603 of described input channel to the.Comprise a plurality of reative cells 630 for the 3rd layer 603, it is communicated with described fluid input channel fluid by through hole.In the embodiment illustrated, reative cell 630 forms with the bottom of layer 603 and links to each other, and in other words, described reative cell is open from the bottom.Therefore, through hole is completely contained in 602 layers.In other embodiments, described through hole can be comprised in 602 and 603 layers among both to be adapted to concrete application.
In some embodiments, microfluidic device can comprise one or more layer according to rotation or the preparation of cast fabrication scheme.For example, the rotation scheme can relate on figuratum dish or mould places polymeric material, and rotates described dish to produce polymeric layer on dish.Illustrative polymers draw together polymethacrylates, polystyrene, polypropylene, polyester, fluorinated polymer, polytetrafluoroethylene (PTFE), Merlon, polysilicon and dimethyl silicone polymer (polydimethylsiloxane, PDMS).Casting program can relate to as the PDMS material is cast on figuratum template or the mould, and this can produce the PMDS layer that can intactly peel or leave behind from the mould.Usually, use the bed thickness of rotation manufacturing technology preparation by the layer ratio of cast manufacturing technology preparation.Elastomer block can comprise one or more cast or the rotating layer of any required combination.
In some embodiments, can make ground floor 603 according to the rotation scheme.For example, PDMS can be placed on the mould that has corresponding to the bossing of multiple required control channel 610.Rotatable described mould is to produce the PDMS thin layer on entire die.After curing, can peel ground floor 601 off and be attached on suitable rigid matrix (as glass, silicon) or the plastics (polystyrene) from mould.Perhaps, ground floor 601 can remain and be attached on the mould.Ground floor 601 can comprise opening, groove or form or limit other spaces of control channel 610 to small part.
In order to generate the second layer 602, can adopt the rotation scheme, PDMS is placed on second mould, described second mould has the bossing corresponding to multiple required fluid input channel.Second mould also can comprise, and for example forms corresponding to the projection or the wavy part of arranging mark in the second layer 602.These are arranged mark and can use in the laser ablation that is used to form through hole 625, so that laser ablation points to described arrangement mark during the ablation processes.Rotatable second mould is to provide PDMS thin layer on whole second mould.The second layer 602 can comprise opening, groove or form or limit other spaces of input channel 620 to small part.In some cases, the second layer 602 can be exposed to the above-mentioned laser ablation of one or many.The ablating laser beam that points to the second layer 602 can form through hole 625.After the second layer 602 fully solidifies, can peel off ground floor 601 from second mould, and it is alignd with ground floor and contact.The second layer and ground floor can be pasted so that two-layer sticking on the rigid matrix.
In order to adopt casting program to produce the 3rd layer 603, PDMS can be cast on the 3rd mould that has corresponding to the bossing of multiple required reative cell 630.After curing, can peel the 3rd layer 603 and align with the second layer 602 and contact off from the 3rd mould.Can with the 3rd layer stick on the second layer so that all three layers stick on the rigid matrix.The material that can make microfluidic device includes but not limited to the combination of elastomer, silicon, glass, metal, polymer, pottery, inorganic material and/or these materials.
Referring to Fig. 1, pressure source 102 is communicated with sealing control line 110 fluids again.Described locking control line comprises the section and the check valve 112 (being also referred to as check-valves (check valve)) of several hereinafter more complete descriptions.Other description about check valve is provided in the international patent application No.PCT/US07/080489 of common unsettled and common transfer (open with international publication number WO 2008/043046A2, its whole disclosures are quoted by integral body and incorporated this paper into).First section 114 from pressure source 102 receive start pressure and can operate in order to close exist the closed array 106 hold valve (containment valve).As understood by the skilled person, array 106 is suitable for carrying out multiple micro-fluid experiment.Thereby can close and hold valve by responding to starting of locking control line 110, make the sample that exists in the reative cell in the array, reagent etc. be retained in the reative cell.The section 114 no check-valves of locking control line 110, this make hold valve remove pressure source 102 start pressure after open again.
Second section 116 of locking control line 110 is in the downstream of check valve 112.Because check valve 112 can be operated in order to stop fluid to flow to section 114 from section 116, starting of pressure source 102 will cause controlling fluid (being generally liquid) by check valve 112 and shut off valve 118a-118e.The fine rule that is associated with locking control line 110 shown in Fig. 1 is represented control line " overhead (flyover) " section, and wherein the design of control line prevents to measure input line 132 (describing more completely hereinafter) and closes after locking control line 110 starts.The representative of the thick line that is associated with valve 118a-118e can be operated in order to close or the valve of inactive manifold 135 (fluid that is provided between a plurality of mensuration input lines 132 flows).Close to stop and measure stream process manifold 135 from a mensuration input line of measuring the input linear flow to other in the pass of valve 118a-118e, this is called stopping using of manifold.
After starting locking control line 110 by pressure source 102, valve 118a-118e can close and remove the locking control line start pressure after, the closed condition that check valve 112 can maintaining valve 118a-118e.118a-118e is opposite with valve, can describedly be opened after starting pressure removing by the valve that holds that section 114 starts.Therefore, the locus of check-valves one or more section that to provide with locking or non-locking behavior be feature in the locking control line.Clear and definite as those skilled in the art, in control line, adopt extra check-valves that the extra section with locking behavior can be provided.
Locking control line 110 also comprises the 3rd section 128, and it has can be operated in order to close from measuring input 130 imports the valve of line 132 to the mensuration of array 106.Measure input 130 and also can be called as the input port.Because the 3rd section 128 is also in the downstream of check valve 112, by pressure source 102 start locking control line 110 can close the valve in the 3rd section 128 and stop measure input line 132 and array 106 with and on flowing between the reative cell of arranging.Start what remove locking control line 110 that the valve of section 128 keeps cutting out behind the pressure.
Second pressure source 104 that is communicated with second control line, 120 fluids is provided, and it is called as interface accumulator (interface accumulator).Described second control line also comprises several segments 121,123 and 125.Section 121 comprises elevated region 122 so that can start separate valves 124 under the situation of not closing mensuration input line 132.Provide in this specification the valve 124 other description of (being called independence or separator valve (independent or decoupling valve) herein), and hereinafter it has more specifically been described.Although use second pressure source 104 to start in specific independence described in Fig. 1 or separator valve 124, this is optional in the present invention.In other embodiments, independence or separator valve 124 can be machinery, static, fluid, electromechanical, thermodynamic, piezoelectricity etc.Therefore, although adopt second pressure source 104 that shows among Fig. 1 to start separator valve 124, this is optional in some embodiments.In addition, although use single second pressure source 104 control to comprise separator valve in interior a plurality of valve groups, other embodiments can adopt that combination that a plurality of pressure source fluids start and static start etc.Those of ordinary skills understand multiple variation, improvement and replacement.
Section 123 comprises the valve 134 that can be operated in order to close in the mensuration input line 132 shown in 65.Owing to only provide 5 valves 134 in the embodiment depicted in fig. 1, the mensuration input line that still can make low order end that starts of valve 134 is held open in the section 123 that is undertaken by pressure source 104.Section 125 comprises check valve 127 and can be operated in order to close the valve 126 of sample input line 142, and described sample input line 142 is communicated with sample input (being the input port) 140 and array 106 fluids.Although should note in Fig. 1, having shown the input of mensuration and sample, the invention is not restricted to have only and measure and the sample input, other inputs are included in the scope of the present invention.
Unidirectional or check-valves 127 are placed on the section 125 of locking control line 120, allow 126 valves to close and keep closing, stop fluid flowing by sample input line dealing array 106 in the section back of exerting pressure.Because section 121 and 123 does not comprise check-valves, cancelling of these sections started and will be caused the unlatching again of valve 134 and 124.
Embodiments more of the present invention provide the asynchronous logic function in microfluidic device 100.For example, owing to use separately and the control line 120 that is independent of control line shown in other 110 starts independence shown in Fig. 1 or separator valve 124, embodiments more of the present invention provide multiple control.In Fig. 1, before starting separator valve 124, locking control line 110 is exerted pressure and can locking be held valve 128.On the other hand, by before holding 102 pairs of locking control lines 110 of accumulator and exerting pressure, close separator valve 124 closing of valve 128 held in prevention thereby start locking control line 120 by interface accumulator 104.In other words, if separator valve 124 cuts out when valve 128 is begun to exert pressure, then the closed condition of separator valve stops institute's applied pressure to arrive valve 128.Therefore, control line 110 with 120 start the order cause different valves to be operated, this provides asynchronous logic.
The multiple control that embodiment of the present invention provides can be " programmed " array 106.For example and since array 106 by a plurality of valves close, locking or the sequential programming opened again, can use array 106 with multiple layout.In embodiments more as herein described, 2 pressure sources shown in the use provide three kinds of different layouts.Adopt check valve and separator valve (it can be considered that piles up in the control valve group) that these multiple layouts are provided.Separator valve can limit the control to valve 128, and described control can be by responding to the realization of starting of locking control line 110, and described separator valve just piles up an example of control valve, and the scope of the invention comprises other layouts.
Use check-valves that the part locking of control line is provided in the predetermined segment of control line.For example, can be by initial control line 110 start locking-valve 118 and 128.Yet starting of initial control line 120 can cause closing of separator valve.The locking that can cause valve 118 of starting of control line 110 does not subsequently still have influence to valve 128.When retentive control line 110 starts, cancel subsequently start control line 120 (for example by discharge interface accumulator applied pressure) can cause separator valve open again after the locking of valve 128.
In microfluidic device shown in Figure 1 100, measure input line 132 for 6 and be divided into 4 input lines separately, provide and passed 24 the mensuration input lines altogether that hold valve 128.In addition, 24 sample input lines 142 have been shown.The concrete numeral of these input lines is an example, and other embodiments provide other concrete input line numbers.For example, in a specific embodiments, schematic diagram shown in Figure 1 has only shown half microfluidic device (for example Zhuan Zhi right side), and the input group that is complementary is arranged in the left side of described device.Those of ordinary skills can understand many changes, improvement and replacement.
Fig. 2 is the simplification sectional view of the check valve of one embodiment of the invention.Fig. 2 B is the simplification top view of check valve shown in Fig. 2 A.Referring to Fig. 2 A, start fluid (being generally liquid) through the 210 inflow check-valves 112/127 that enter the mouth, and flow through chamber 220 through through hole 215.Mention or remain on the position of basic horizontal by the mobile film 230 that makes that starts fluid, make and start fluid and 210 from left to right pass through inlet.Starting fluid via through holes 240a-240d and through hole 242 flows through chamber 220 and flows out from exporting 250.Therefore, in the embodiment illustrated,, an input through hole and 5 output through holes are arranged for each check valve 6 through holes altogether.As shown in the top view of Fig. 2 B, provide extra structural detail and assembly according to specific design.
When fluid pressure was started in removal, film 230 was collapsed upon on the layer 260, stoped by inlet 210 and refluxed.Therefore, check valve 124/127 provides from entering the mouth 210 to the flowing of outlet 250, but stops reverse flowing.Being described in about other of check valve in above-mentioned application of quoting of common unsettled and common transfer provides.
Embodiments more of the present invention provide the microfluidic device that is integrated with unidirectional or check-valves.In one embodiment, described microfluidic device comprises first valve and second valve.Usually, described first valve is one that forms in a plurality of valves of the first valve group, and described second valve is one that forms in a plurality of valves of the second valve group.The control line that is communicated with described first valve and the described second valve fluid provides with pressure source as the part of microfluidic device, and described pressure source is communicated with described control line fluid.Referring to Fig. 1, by exerting pressure and start control line 110, manifold 135 is stopped using thereby cause valve 615 in the array 106 and valve 118a-118e to start to holding accumulator 102.Described microfluidic device also comprises the check valve in the control line that is arranged between described pressure source and described second valve.For example, the valve of originally opening 112 is on the control line 110 that holds between accumulator 102 and the valve 118a-118e.Reduce or remove pressure on the control line 110 after check-valves 112 locking of valve 118a-118e is provided, and the valve (non-locking-valve) that exists in the array 106 can be opened again and makes sample flow into reative cell.Therefore, by adopting the valve of predetermined number in the check-valves locking microfluidic device, fluid flows in the irrealizable microfluidic device in the common design the control and the separation of sample are provided.
In the embodiment shown in Fig. 1 and the 6A, valve 615 is set to the reative cell of spaced apart on microfluidic device, and valve 118a-118e be set to with measure input port 130 be connected (coupled) six lines first fluid input line with import line with second fluid of measuring six lines that the input port is connected and separate mutually.As more complete in this manual description, thereby close the ability that manifold 135 is stopped using with locking-valve 118a-118e microfluidic device can be programmed in many ways, this flows the fluid of importing in the line 132 can the separated or merging according to concrete application.
Except check-valves being integrated in the microfluidic device with array layout, some designs provide separate valves, are also referred to as separator valve.For example, be independent of other valves in the device and controlled separator valve can be operated in order to stoping starting of one or more valve, described one or more valve control is mobile by the fluid of the fluid input line that is connected with the reative cell that exists in the microfluidic device.For example, can before starting control line 110, close separator valve 124, thereby stop valve 128 to respond to closing that control line 110 starts.
Figure 10 is the rough schematic view of the programmable microfluidic device of one embodiment of the invention.Element shown in Figure 10 can provide in carrier or provide in being attached at the microfluidic device of carrier as shown in figure 11.As shown in figure 10, be loaded on the profile of the microfluidic device on the carrier by dotted line 1050 expressions.Referring to Figure 10, two emission sources (vent source) are provided at the opposite side of microfluidic device.The common emission source that provides as the part of carrier can not adopt in all embodiments.48 sample input lines 1010 above Figure 10, have been shown, wherein arranged 24 samples input lines and to array 106 left side sampling, arranged 24 samples input lines on the microfluidic device right side and array 106 right side sampling in the left side of microfluidic device.Article 48, sample wire provides in carrier and usually by forming in microfluidic device and being communicated with array 106 fluids with through hole 1030 that the sample wire end aligns.106 sample input line 142 is provided in microfluidic device from through hole 1030 to array.Article 48, sample wire is by the pressurization of UP source (common pressure source) (not shown), and the sample of the carrier that described pressure source can be placed on it with microfluidic device partly is connected.Therefore, the sample of 48 sample ports can be written into microfluidic device and push array 106 through sample input line.Referring to Fig. 1, for the sake of clarity shown the right side part of array 106, wherein 24 of Figure 10 right side part sample input lines are shown as sample input line 142.Therefore, embodiments more of the present invention provide 48 sample input lines with 48 different samples of as many as.Other implement to provide different numbers according to concrete the application.
Shown control line 1004 in Figure 10, it is connected with Fig. 1 interface displayed accumulator 104.In addition, shown that in Figure 10 (check valve, CV), this provides the one-way fluid flow through control line to several check-valves in microfluidic device.Referring to the bottom of array 106, for the control line that is connected to this part of array provides additional C V.Provide the aquation line in the outside of array 106.These aquation lines provide the source for the aquation fluid, and the effect of aquation fluid is the evaporation that reduces or prevent fluid in the array.
48 input lines that show in Figure 10 bottom comprise 12 and measure input line (6 in array 106 left sides, 6 on array 106 right sides).Article 12, measure line by the pressurization of UP source (not shown), the mensuration of the carrier that described pressure source can be placed on it with microfluidic device partly is connected.Therefore, 12 mensuration of measuring mouth can be written into microfluidic device and import line after measured and push 2 arrays 106.Referring to Fig. 1, for the sake of clarity show right array 106, it has 6 sample input lines that are shown as Figure 10 right side part of measuring input line 132.Therefore, embodiments more of the present invention provide and have had 12 different 12 mensuration input lines measuring of as many as.Measure the input line and pass through between two arrays 106 of central authorities of microfluidic device, 6 lines are branched off into 106,6 lines of right array and are branched off into left array 106.Shown in the use of manifold can provide and be less than 12 mensuration, for example, few to single mensuration.Other implement to provide different numbers according to concrete the application.
In other 36 shown in Figure 10 bottom input lines, 2 lines are used for aquation, and 34 is untapped in this concrete enforcement.Measuring input line 1020 provides the input of the single input line that enters array 106, is branched off into the many input lines (for example, 4 lines) that enter mensuration 106 and measure input line 1022.
Fig. 6 A has shown the example of 3 inputs of every sample line, but in the embodiment depicted in fig. 10,11 input lines is arranged.In addition, in Fig. 6 A, 2 control lines are presented among Fig. 6 A, but in the embodiment depicted in fig. 10, have 71 control lines can be operated to form 70 reative cells/input line.Therefore, in the present embodiment, 770 reative cells (11 input line * 70 reative cell) are arranged for each sample.Along with 48 samples are dispensed into 770 reative cells, provide highdensity integrated fluid loop (integrated fluidic circuit, IFC).In one embodiment, microfluidic device can one-time detection reach 48 independently samples.Each of described nearly 48 samples is assigned with into 770 reative cell independent groups, thereby sends the synchronous digital PCR reaction as ading up to 36,960.In another embodiment, remove some programmability of microfluidic device, then, increased the reaction number of chambers/sample for identical device area and size of components.For example, an embodiment is used nearly 48 samples, and it has 814 reative cell/samples.
Use can obtain the product from the assignee of the present invention, can finish whole digital pcr process in 4 hours being less than.In addition, microfluidic device as herein described is the part of complete gene alaysis system.This system can comprise controller, the BioMark of microfluidic device, microfluidic device TMSystem or integral type thermal cycler, EP1 read instrument and related software.The distribution design compatibility of microfluidic device as herein described and finished product reagent and micropore form.Those of ordinary skills understand many changes, improvement and replacement.
Figure 11 is the rough schematic view of the microfluidic device of one embodiment of the invention.Described microfluid system comprises carrier 1100 and the microfluidic device 1108 that is loaded on the described carrier.Microfluidic device 1108 has been integrated the described element of Fig. 1.Described carrier comprises a plurality of first input ports or the hole 1105 that is arranged on platform (bank) 1106a.Described carrier also has a plurality of first input lines 1115 (for example, 48 input lines) that are communicated with a plurality of first input port fluids.Around in described a plurality of first input ports outer peripheral edges are arranged, allow to be pressurizeed in described first input port with the UP source.In an exemplary application, 48 samples are written into first input port, from the top of carrier described mouth being exerted pressure causes 48 samples to be pushed over a plurality of first input lines and finally enters fluid line on the microfluidic device.
Described carrier also comprises the second input line 1115 of a plurality of second input ports 1105 of being arranged on the platform 1106b and a plurality of correspondences.Be similar to a plurality of first input ports, described a plurality of second input ports are centered on by outer peripheral edges, make the fluid that is dispensed into second input port can be pushed over go forward side by side fluid line in the microfluidic device of the second input line.
For at described carrier with place and provide fluid to be communicated with between the microfluidic device on the described carrier, microfluidic device has the through hole 1114 that forms in the microfluidic device bottom, and it aligns with the described first input line and described second end portion of importing line.The fluid that flows through the input line passes through hole and upwards enters the fluid line that microfluidic device provides.As shown in figure 10, the fluid line that connects described sample input port enters the offside of microfluidic device through the through hole 1030 along microfluidic device left side and right side arranged vertical among the figure, and flows to the central authorities of microfluidic device.In Fig. 6 A, but the input line 620 that horizontal flow is crossed among these sample input line corresponding diagram 6A.Use this 48 sample input lines, can in the reative cell of the every row of microfluidic device, fill different samples.
Also provide through hole 1040 and its to align in the bottom of microfluidic device with a plurality of second input lines 1020.As shown in figure 10, the fluid (as measuring) that is written into second input port flows through the second input line 1020, enters manifold 1060 through through hole 1040.Described microfluidic device comprise can with the crew-served valve of described manifold, be provided two reative cells in the array to cause 1 fluid (sample or mensuration) or 12 different fluids.If being unlocked and importing fluid, the single input fluid of application need, manifold flow to all fluid lines that pass through through the array central vertical among Figure 10.Fluid line the time is branching out through array central authorities, and finally the through hole 1030 that flows through array side along with them finally flows through reative cell.Referring to Fig. 6 A, these measure input lines can be corresponding under reative cell the input line 620 of process.In this example, the mobile of left array side is from central authorities to the left side, and the mobile of right array is to the right side from central authorities.
Perhaps, can seal manifold 135 to introduce 12 different fluid (for example measuring) (being 6 different mensuration of each array) of waiting to introduce reative cell.As shown in figure 10, fluid is perpendicular flow between array, and branch goes out then, and wherein each of 12 fluids flows among many rows of array, flatly from the through hole 1030 of central authorities through arrival edge vicinity.
Providing acute build up of pressure device 1106c and 1106d on the carrier so that can start control line and the check-valves that exists on the microfluidic device.With with the similar mode of fluid line, the control line 1002 in the carrier and 1004 is communicated with control line 110 and 120 fluids on the microfluidic device by the through hole that forms on the microfluidic device.Therefore, although Fig. 1 shown the interface of carrier and held control line in accumulator and the microfluidic device, should understand this schematic diagram and be and be the sketch for the purpose of clear and convenient.Other details relevant with the carrier that can place microfluidic device thereon provide in U.S. Patent Application Publication No.2005/0214173, and its disclosure is quoted by integral body for all purposes and incorporated this paper into.
Carrier 1100 has the acute build up of pressure device hole 1101 and 1102 of integration, has the dry hole 1103,1104 that is used to hold valve in its each, and described valve is preferably and covers the check-valves that links to each other.Carrier 1100 also comprises one or more hole platform 1106a, b, c and d, its each have and be positioned at one or more hole 1105 (being also referred to as the input port).The hole 1105 of each carrier 1100 has from the hole passage that 1105 guiding place the microfluidic device 1108 of carrier positions 1107.Hole platform 1106c and 1106d are generally used for providing the pressure that is used for starting existing control line in the microfluidic device 1108.The fluid line of control fluid is provided, valve or other control appliances that exists in the hole of Kong Taizhong and the microfluidic device is connected.Microfluidic device is preferably the elastomer block that is formed by two-layer or more multi-layered elastomeric material, and it has the groove or the passage of the little manufacturing that forms therein.
The one or more passages that are communicated with one or more through hole 1114 fluids are arranged in the described microfluidic device, it provides the fluid between the passage in passage and the carrier in the microfluidic device to be communicated with then, hole 1105 among its pilot hole row 1106a-d then, thus provide the fluid between the passage in hole 1105 and the microfluidic device 1108 in the carrier 1100 to be communicated with.Accumulator bore top 1109 and 1110 is connected to accumulator bore 1101 and 1102 to form accumulator chamber 1115 and 1116.Accumulator bore top 1109 and 1110 comprises valve 1112 and 1111 respectively, and it is preferably under pressure and gas is introduced accumulator chamber 1115 and 1116 and keep the check-valves of gas.In the time of in being present in accumulator chamber 1115 and 1116, valve 1111 and 1112 is positioned at dry hole 1102 and 1104 inside so that liquid does not contact valve 111 and 1112.Preferably by pushing mechanically open valve 1111 and 1112 such as thin slice (shave), pin in the preferred check-valves, its power of closing automatically that overcomes check-valves is with from accumulator chamber release pressure, thereby reduces contained fluid pressure in the accumulator chamber.
Carrier 1100 and its associated component can be by the polymer manufacturings, as polypropylene, PEF, Merlon, high density polyethylene (HDPE), polytetrafluoroethylene PTFE or Teflon (R), glass, quartz, metal (for example aluminium), transparent material, polysilicon etc.Accumulator bore top 1109 and 1110 also comprises the adjustment screw, and it is removable to introduce or to remove gas or liquid from accumulator chamber 1115 and 1116.Preferably, can start valve 1112 and 1111 with fluid pressure in release accumulator chamber 1115 and 1116, otherwise fluid pressure retains wherein then.Use notch 1117 to help microfluidic device is placed in other instruments suitably, for example, be used to operate or analyze microfluidic device or the instrument of the reaction wherein carried out.
Fig. 3 is the simplified flow chart of the method for display operation microfluidic device according to an embodiment of the invention.48 panel layouts of digital array are provided in the embodiment depicted in fig. 3.Pressure source 102 (holding accumulator) is applied first pressure, and this has caused starting of locking control line 110 (310).In a specific embodiment, described first pressure is 30psi.In other embodiments, adopt other pressure according to concrete application.Referring to Fig. 1, all the valve (not shown)s in the array 106 can be owing to the pressure of section 114 is closed.Pressure is applied to section 116 by check valve 112, and this can cause closing of valve 118a-118e and locking.The overhead part of section 116 can stop measures the input line owing to starting of section 116 closed.Close and close the input fluid through the flowing of manifold 135 in the pass of valve 118, to measure the input lines separate thereby will be connected to 6 of measuring input 130.
Should notice that the working pressure source 104 in control stream shown in Figure 3.Therefore, there are not starting of control line 120 or closing of separator valve 124.Therefore, respond to starting of locking control line 110, hold valve 128 and close also locking.Therefore, by hold accumulator apply first pressure (for example, 30psi) after, described array valve cuts out and valve 118 and 128 cuts out and locking.
First pressure (312) that reduces pressure source 102 is opened array valve (not shown) again.In some embodiments, in process (312), remove pressure so that institute's applied pressure is zero.As noted above, valve 118 and 128 can keep locking in off position.Supply is written into reative cell in (314) array from the sample of sample input or mouthfuls 140 by sample input line 142.The design of chip provides by sample input line 142, measures input line 132 or both loadings.Because in this embodiment, measure the input line and close by locking-valve 128, load through sample input line and carry out.After finishing sample and being written into, pressure source 102 is applied second pressure (316) again, thereby close the closed array valve and separate sample in the described reative cell.In some embodiments, second pressure equal first pressure (for example, 30psi), although this is optional in the present invention.Other pressure that are suitable for closing the closed array valve within the scope of the present invention.In addition, although removed first pressure in some embodiments, this is optional in the present invention, still provides the sufficient fluid of load sample to flow because some embodiments pressure can be reduced to nonzero value.
Therefore, Fig. 3 embodiments shown provides the loading (that is the input of the sample shown in 24 line * device both sides) of 48 samples.As discussed below, programmable digital array as herein described also provides other sample layouts.
Should understand concrete steps shown in Figure 3 provides a kind of ad hoc approach of the microfluidic device of operation one embodiment of the invention.Other sequence of steps also can be implemented according to some alternate embodiments.For example, alternate embodiments more of the present invention can be implemented step listed above with different orders.In addition, single step shown in Figure 3 can comprise a plurality of substeps, and it can be implemented with multiple order according to the requirement of single step.In addition, can or remove other step according to concrete application interpolation.Those of ordinary skills can understand many changes, improvement and replacement.
Fig. 4 is the simplified flow chart of the method for a kind of operation of demonstration microfluidic device according to another embodiment of the invention.Different with described 48 panel layouts of Fig. 3, the single sided board layout is provided in the embodiment depicted in fig. 4.Pressure source 104 is applied first pressure (410), and this causes starting of locking control line 120.Pressure can by check valve 127 apply with close with locking section 125 in valve 126.Therefore, in this layout, do not have flowing through sample input line 142.Overhead section 122 can transmit controlled pressure to close separator valve 124.
Section 123 can be passed to the controlled pressure in the locking control line 120 with 6 and measure 5 valves that are associated 134 in the input line 132.Therefore, valve 134 will be closed, and making flows only imports line through the mensuration of low order end.
Traditional thread binding year single sample (412) of mensuration input through low order end.Because the section 116 of locking control line 110 is not started in this layout, manifold 135 is opened, and makes that contained single sample is provided to whole sample input lines 132 in the input of low order end sample.Therefore, use the single sample of importing from the low order end sample determination to import after measured and load described array.Acute build up of pressure device 102 is applied second pressure, start locking control line 110 and close the array valve that links to each other with section 114.Therefore, use different control stream, it is possible using the multiple layout of single programmable digital array, for example, and the single sided board layout.
Should understand concrete steps shown in Figure 4 provides a kind of particular methods of operation of microfluidic device according to another embodiment of the invention.Other sequence of steps also can be implemented according to alternate embodiment.For example, alternate embodiments more of the present invention can be implemented step listed above with different orders.In addition, single step shown in Figure 4 can comprise a plurality of substeps, and it can be implemented with multiple order according to the needs of single step.In addition, can or remove other step according to concrete application interpolation.Those of ordinary skills can understand many changes, improvement and replacement.
Fig. 5 is the simplified flow chart of method that shows the microfluidic device of a kind of operation another embodiment according to the present invention.12 panel layouts of digital array are provided in the embodiment depicted in fig. 5.Pressure source 104 is applied first pressure (510).In a specific embodiment, described first pressure is 45psi, although this is optional in the present invention, can the pressure that adopts other to be fit to.As the discussion of the process (410) of relevant Fig. 4, start locking control line 120.By check valve 127 exert pressure with close and locking section 125 in valve 126.Therefore, in this layout, do not have flowing through sample input line 142.Overhead section 122 can transmit described controlled pressure to close separator valve 124.As discussed below, the Temporarily Closed of separator valve can stop and holds valve 128 and close under the effect of starting of subsequently control line 110.Section 123 can be passed to the controlled pressure of locking control line 120 with 6 and measure 5 valves that are associated 134 in the input line 132.Therefore, valve 134 can be by temporary close.
Pressure source 102 is applied second pressure (512), start locking control line 110.In a specific embodiment, described second pressure is 30psi, less than first pressure (for example, 45psi).As discussed below, described second pressure is enough to close predetermined valve and is not open at the separator valve 124 of closing in the process (510).Can use other pressure according to concrete application.The valve that closes in the closed array 106 is understood in starting of section 114.Pressure can be applied to section 116 through check valve 112, and this can cause closing of valve 118a-118e and locking.The overhead part of section 116 can stop measures the input line owing to starting of section 116 closed.Closing and close the input fluid through the flowing of manifold 135 in the pass of valve 118, thereby will measure and import lines each will be separated with 6 that measure that input 130 be connected.
Start differently with control line 102 initial, initial the starting of control line 104 closed separator valve, thereby programmable sequences is provided, and wherein starting of control line 102 do not cause and measure valve 128 lockings that input line 132 is associated.This multiple control characteristic provides the asynchronous logic function that adopts some embodiments of the present invention, uses microfluidic device commonly used can not obtain this characteristic.
The pressure that is applied to pressure source 102 is reduced to for example zero (514).Because section 116 is in check valve 112 downstreams, to cancel at section 116 and to start back valve 118 and continue to be latched in the closed position, this stops 6 fluids of measuring between the input line to flow.The valve of opening in the described array is started in cancelling of section 114, makes load sample in loading process subsequently.As discussed above, valve 128 is non-latching in process (512).
The pressure that is applied to pressure source 104 is reduced to for example zero (516).Pressure by reducing section 123 is open valve 134 again.As discussed above, use mensuration input 130 to come to provide input as the reative cell in the array.The pressure in the overhead section 122 reopens separator valve 124 in the section 121 by reducing.Owing to control line 110 is not exerted pressure, so valve 128 remains on opening in this stage of programming process.Although that is showed is zero for pressure is reduced to, these concrete pressure are not essential to the invention, the pressure that can adopt other to be fit to.
From 6 mensuration inputs, be written into sample (518).By since before the functionalization of going of the manifold 135 that causes of the locking of valve 118 keep measuring separation between the input.Therefore, in this layout, 12 panel layouts (6 2 sides of measuring input * device) are provided.Pressure source 102 is applied the 3rd pressure (520) to close the array valve that is associated with section 114.In addition, because separator valve 124 reopened, so valve 128 is cut out by locking.
Should understand concrete steps shown in Figure 5 provides a kind of concrete grammar of the microfluidic device of another embodiment of operation the present invention.Other sequence of steps also can be implemented according to alternate embodiment.For example, alternate embodiment of the present invention can be implemented step listed above with different orders.In addition, single step shown in Figure 5 can comprise a plurality of substeps, and it can be implemented with multiple order according to single step.In addition, can or remove other step according to concrete application interpolation.Those of ordinary skills can understand many changes, improvement and replacement.
Table 1 provides starting pressure and being written into process of one of a plurality of possibility panel layouts of programmable digital array microfluidic device as herein described.Pressure source 102 and 104 applied pressures are described with exemplary pressure, although other press packet that are fit to are contained in the scope of embodiment of the present invention.
Programming step 48 panel layouts 1 panel layout 12 panel layouts
1 102＝30psi 104＝30psi 104＝45psi
2 102＝0psi Be written into sample 102＝30psi
3 Be written into sample 102＝30psi 102＝0psi
4 102＝30psi 104＝0psi
5 Be written into sample
6 102＝30psi
Table 1
The integration of check valve and separator valve and digital array provides the multi-user who uses single high-density digital array chip the layout of programming.Therefore, embodiments more of the present invention provide the saving of cost and the raising of experiment flux.Serial described herein or asynchronous logic provide the enhanced device function, and this is because the state of particular valve not only depends on starting of the control line that is communicated with the particular valve fluid, also depends on the state of the separator valve that independently control line started.Therefore, the order of pressurization has determined the logic output of panel layout.Therefore, embodiments more of the present invention provide device layout and the function that adopts conventional microfluidic device not obtain.
According to one embodiment of the invention, provide the method that the microfluidic device with a plurality of control lines is provided with.Serial or asynchronous logic have been implemented according to this method.Described method comprises starts first control line and valve is placed first state.Valve (can be in a plurality of valves) can respond to described first the starting of control line and place closed condition.For example, separator valve 124 can respond to exerting pressure and the pressurization of control line 120 and cutting out to interface accumulator 104.More specifically describe as more complete description in this specification and following, because separator valve is placed in valve 128 and holds between the accumulator 102, separator valve can provide multiple control function, and this provides the serial logic operation.
Described method also comprises, after valve is placed first state, starts and can be operated in order to the valve group is placed second control line of second state.The situation that valve is in first state (promptly cutting out) stops the valve group to be placed in second state (for example, closed condition).As shown in Figure 1, closing separator valve 124 before starting control line 110 stops valve 128 to respond to starting of control line 110 and close (and locking).Therefore, embodiments more of the present invention provide the serial logic function, and wherein the valve order of starting has produced the different final results that are selected from several end-state.In this example, initial the starting of control line 110 causes closing and locking of valve 128.On the other hand, initial the starting of control line 120 causes closing of separator valve 124, and it stops closing and locking of valve 128 when remaining on closed condition.Although Fig. 1 has shown the control line 110 and 120 of exerting pressure and being started to pressure source 102 and 104 independently, other start method and comprise within the scope of the present invention.
According to another embodiment of the invention, provide and adopted microfluidic device to carry out the another kind of method of serial logic.The method that microfluidic device with a plurality of control lines is provided with comprises by starting first control line sets up first state of microfluidic device, then starts second control line.First state of microfluidic device can comprise that the valve that starts that responds to many control lines is set to open or closed condition.For example, as mentioned above, before starting control line 110, start control line 120 and cause closing of separator valve 124.Therefore, valve 128 does not respond to starting of control line 110 and locking is closed.
Described method also comprises by starting second control line sets up second state of microfluidic device, then starts described first control line.The programmable features of microfluidic device as herein described provides and has depended on that valve starts the different end-state of the microfluidic device of order.Therefore, in this example, before starting control line 120, start control line 110 and cause closing and locking and closing of separator valve 124 thereafter of valve 128.It is adiaphorous closing separator valve behind locking-valve 128, because check-valves 112 maintenances to the constant pressure of the control line of check valve downstream, stop the state that influences microfluidic device of closing of separator valve.
Implement the example of serial logic as microfluidic device, first state of microfluidic device can comprise close or the first input line group of blocked state (for example valve 126 blocked input line 142) and is communicated with a plurality of reative cell fluids in the described microfluidic device second import line group (for example importing line 132).Described second state can comprise the described first input line group (for example importing line 142) that is communicated with a plurality of reative cell fluids in the described microfluidic device, and closes or the second input line group of blocked state (for example valve 128 blocked input line 132).Second state also can comprise the valve group (for example valve 118a-118e) of closed condition, thereby stops fluid through connecting the manifold flow of the second input line group.
Started by control line 120 although should note separator valve, other embodiments can adopt the another kind of separator valve that is started by control line 110, with as shown in the substituting or replenishing of separator valve.Therefore, for example, overhead route can lead to extra separation valve between interface accumulator 104 and check-valves 127 from holding accumulator 102.Can stop valve 126 to respond to starting of control line 120 and close and locking before the section 125 of the control line that is connected to interface accumulator 104 is exerted pressure, closing this extra separator valve.Therefore, although Fig. 1 has shown concrete enforcement, the invention is not restricted to this concrete enforcement, other microfluidic device designs within the scope of the present invention.
In addition, although embodiment shown in Figure 1 and replaceability geometric shape discussed above have adopted by one in two control lines and exerted pressure and provide two levels of multiple control with the separator valve that it starts, embodiment of the present invention provide other levels of multiple control.For example, other separate valves (for example, static starts) can be integrated in the design of microfluidic device, so that the control of the 3rd level to be provided.Except the starting of the control line shown in 2, to this other separate valves initial start can the drive unit state logic output.
Fig. 7 has shown the method that the programmable microfluidic device of one embodiment of the invention is programmed.In the embodiment depicted in fig. 7, the reative cell that described microfluidic device is programmed for described microfluidic device is separated into 48 independently panels.The concrete number of panel depends on the actual design of microfluidic device, and can be different numbers in the embodiment of replaceability, as 12 or 192 panels.The reative cell of each separate panels can be filled different samples, provides in 48 different samples of as many as to detect in the present embodiment.
As shown in fig. 1, microfluidic device has reaction site or the reative cell array that is arranged in the array 106.Described reaction site is communicated with the first input line group 132 and the second input line group, 142 fluids.Described method comprises that starting (710) can be operated in order to block the first valve group of (712) first input line groups.For example, the first valve group is a valve 128, is also referred to as can be operated in order to stop or to close the valve that holds of input fluid line 132.For example by stoping fluid to flow by importing line 132 to holding accumulator 102 shut off valve 128 of exerting pressure.Because existence unidirectional or check-valves 112 makes control line 110 become the locking control line, after holding valve 128 and closing, their keep closing.
Described method comprises that also starting (714) can be operated in order to separate the second valve group of (716) described reaction site.Shown in Fig. 6 B, reative cell 630 is limited in the elastomer layer 603 of microfluidic device.The described second valve group is arranged in the array, and is shown as valve 615 in Fig. 6 A.Except that the first valve group 128 and the second valve group 615, the 3rd valve group 118a-118e and the described first valve group are started simultaneously.It is locking after closing that the existence of check-valves 112 also makes valve 118a-118e.In the embodiment depicted in fig. 1, described first valve group and the described second valve group respond to control line 110 applied pressures are started simultaneously or close.
After described first, second closed with the 3rd valve group, the described second valve group was cancelled and is started (718), for example, and by reducing or eliminating by 102 pairs of control line 110 applied pressures of pressure source.The unlatching that causes described valve is started in cancelling of described valve, the input line that this flows through fluid to link to each other with valve.A plurality of samples are written into reaction site (720) by the second input line group 142.In Fig. 1, sample is included in sample input or the mouth 140.The both sides of described microfluidic device respectively comprise 24 different input ports, and totally 48 input ports are provided, although this concrete numeral can change according to the specific design of microfluidic device.Sample port can comprise 48 different samples or when providing single sample in a plurality of input ports, can comprise sum sample still less.Therefore, although table 1 relates to 48 panel layouts, in fact the programmable microfluidic device provides independently panel of 48 of as many as according to the sample that provides at 48 sample ports.After sample is written into reaction site, start the second valve group so that reative cell separate (722) for the second time.
Should understand concrete steps shown in Figure 7 a kind of concrete grammar that the microfluidic device of one embodiment of the invention is programmed is provided.Also can implementing of other sequence of steps according to alternate embodiment.For example, alternate embodiment of the present invention can be implemented step listed above with different orders.In addition, single step shown in Figure 7 can comprise a plurality of substeps, and it can be implemented with multiple order according to single step.In addition, can or remove other step according to concrete application interpolation.Those of ordinary skills can understand many changes, improvement and replacement.
Fig. 8 has shown the method that the programmable microfluidic device of another embodiment of the invention is programmed.The method that Fig. 8 shows provides carries out method of operating to having the programmable microfluidic device that single panel and all reative cells comprise same sample.As shown in Figure 1, described programmable microfluidic device has the reaction site array that is communicated with the first input line group and the second input line group fluid.In exemplary shown in Figure 8, the described first input line group is and the sample input 140 input fluid lines 142 that are connected that the described second input line group is and measures the input 130 input fluid lines 132 that are connected.The first input line group started be closed position (810), block (promptly closing) (812) of flowing through the first input line group.Therefore, in an example, 104 pressurizations cause closing of valve 126 to the interface accumulator, stop fluid to flow through input line 142.The existence of check-valves 127 causes the first valve group to be latched in the closed position.
Described method comprises that also starting (814) can be operated the second valve group of importing the first of line group subgroups in order to obstruction (816) second.Referring to Fig. 1, the first of the described second input line group is the part of the input line 132 under the valve 134, and the described second valve group is a valve 134, can operate it to block or to close with 6 mensuration and import 130 6 relevant 5 of importing in the lines 132.Therefore, in the present embodiment, the subgroup of input line is 5 in 6 input lines, adopts the mensuration input port (being connected with the input line of low order end) of low order end shown in Figure 1 that the sample that is written into reative cell or site is provided.Be not included in the subgroup input line (being the input line of low order end) for open and be used for as described in hereinafter complete, sample being written into reative cell.In this concrete enforcement, do not use other to measure the input port.In the embodiment depicted in fig. 1, accumulator 104 applied pressures in interface start valve 126 and valve 134 simultaneously.
Referring to Fig. 1, do not implement starting of the valve that undertaken by the use of control line 110.Therefore, the mensuration input port that sample is written into low order end can cause sample flow to cross section 116 and flow to the second portion (i.e. the part of the input line 132 on valve 134) of the described second input line group by manifold 135.Sample is written in the reative cell or site in the array 106 (818) by the described second input line group.Should note by make manifold stop using (promptly being prevented from) through starting of the section 116 of uninflated control line 110 in the present embodiment through the mobile of manifold.Therefore, manifold is opened and is connected all 6 input fluid lines in the preliminary sectors of the second portion of input line 132.Through manifold 135, sample can be assigned to the component of input line 132, finally flows through 24 and imports line, passes the valve 128 of unlatching.Therefore, single sample is offered all reative cells in the array 106.In order to separate the sample in the reative cell (822), start the 3rd valve group (820), example as shown in Figure 6A hold valve 615.Holding 102 pairs of control lines 110 of accumulator by use exerts pressure the pressure that closes the valve 615 in the closed array 106 can be provided.
Should understand concrete steps shown in Figure 8 a kind of concrete grammar that the microfluidic device of another embodiment of the invention is programmed is provided.Other sequence of steps also can be implemented according to alternate embodiment.For example, alternate embodiment of the present invention can be implemented step listed above with different orders.In addition, single step shown in Figure 8 can comprise a plurality of substeps, and it can be implemented with multiple order according to single step.In addition, can or remove other step according to concrete application interpolation.Those of ordinary skills can understand many changes, improvement and replacement.
Fig. 9 has shown the method for simplifying of programming to the microfluidic device of another embodiment of the present invention.Described programmable microfluidic device has the reaction site array that is communicated with the first input line group, the second input line group and the manifold fluid that is connected the described second input line group.Described method comprises that starting (910) can be operated in order to block the first valve group of (912) first input line groups.Referring to Fig. 1, the first valve group can be the valve 126 in check-valves 127 downstreams.The existence of check-valves 127 makes valve 126 be latched in the closed position after starting.Start the second valve group (914) to block the first (916) of the second input line group subgroup.The pressure that control line 120 is applied from interface accumulator 104 can cause starting of valve 126 and valve 134.
Referring to Fig. 1, the first of the described second input line group is input line 132 parts under the valve 134, and the second valve group is a valve 134, and it can be operated in order to block or to close with 6 and measure 65 of importing in the lines that input 130 is connected.Therefore, in the present embodiment, the subgroup of input line is 5 in 6 input lines.Because valve 134 is not by unidirectional valve blocking, when reopening these valves, the first that fluid can flow through all 6 input lines 132 flows to manifold 135.In the embodiment depicted in fig. 1, the pressure that applies from interface accumulator 104 starts valve 126 and valve 132 simultaneously.
Described method comprises that also starting (918) can be operated the 3rd valve group of using so that manifold is stopped using (920).In an exemplary, the pressure source (for example, holding accumulator 102) that comprises being communicated with the second valve group fluid that starts of the 3rd valve group applies predetermined pressure.Referring to Fig. 1, start valve 118a-118e by the section 116 of control line 110 is exerted pressure and to block through the flowing of manifold, so that the separation to the fluid of the input line 132 of flowing through to be provided.The existence of check-valves 112 can cause valve 118a-118e to be latched in the closed position.As following more complete description, inactive 12 samples (6 samples of the every side of microfluidic device) that make of manifold 135 can be written into reative cell from 6 mensuration input ports 130.Discussed in the embodiment as Fig. 7, be less than 12 samples by identical sample being offered to be written into more than the input of one mensuration.Those skilled in the art will appreciate that many changes, improvement and replacement.
Referring to Fig. 1, should notice that initial the starting of control line 120 not only causes closing and locking of valve 126 and valve 134 before starting control line 110 again, also cause closing of separator valve 124.Because separator valve is arranged in and holds between accumulator 102 and the valve 128, the closing of separator valve can stop valve 128 to respond to control line 110 to start and close.In the embodiment depicted in fig. 9, the pressure that is used to start control line 120 is predetermined pressure, and it is enough to stop and causes starting of control line 110 that valve 128 cuts out.As shown in table 1, the pressure of 45psi is enough to when control line 110 being applied the pressure of 30psi valve 128 be remained on the enable possition.Concrete value 45psi and 30psi are not essential to the invention, can adopt other pressure to implement this multiple control function.
Described method also comprises cancelling starts the second valve group (922).Usually, reduce for example to cancel by first pressure that will be applied to first pressure source and start the second valve group, so that it takes the enable possition to 0psi.The reduction that imposes on the pressure of control line makes the flexible membrane that is deflected into described input fluid line return the position of not deflection substantially, makes the mobile recovery of fluid of the input fluid line of flowing through.Behind open valve 134 again, the sample that provides in measuring input 130 is written into reaction site (924).As shown in Figure 1, each first's (for example part of the input line 134 under valve 134) of the second input line group is communicated with mensuration input port fluid, and described mensuration input port is set to receive one of a plurality of samples.Sample flow enters reaction site through first and second parts of the second input line group.Shown in Fig. 6 A and 6B, described reaction site can a plurality of reative cells form form, wherein each reative cell is communicated with by the second portion fluid of through hole with input fluid line 132, described through hole pass the input fluid line arrive described reative cell, described reative cell can be arranged in contain fluid line the layer on the elastomer microfluidic device layers in.
As shown in Figure 1, the part that is positioned at the input line 132 on the valve 134 is branched off into four input line groups, and it is communicated with reaction site or chamber fluid in the array 106.Provide 6 different samples by measuring in the input port, can in array, define 12 panels (because Fig. 1 has only shown first side of microfluidic device) altogether at each.As it will be apparent to those skilled in the art, in more than one input port, use a sample can cause producing and be less than 12 panels.In order to separate reaction site, start the 4th valve group (926), for example valve 615.
In some implementations, described method also comprises starts valve to stop closing of the 5th valve group, wherein can operate the 5th valve group to close the second portion of the second input line group.As shown in Figure 1, closing of separator valve 124 stops valve 128 to respond to starting of control line 110 and closes.In the embodiment depicted in fig. 9, the pressure that is applied to the 45psi of control line 120 when initial causes closing of separator valve, makes the second portion of importing line 132 be held open the while manifold and stops using by closing of valve 118a-118e.Separator valve can start or can otherwise start by control line is exerted pressure, and includes but not limited to machinery, static etc.In the application of use, can use single control line to start a plurality of valves and/or valve group simultaneously based on the control line of pressure.Therefore, when to 104 pressurizations of interface accumulator, separator valve 124 and valve 126 and valve 134 respond to starting of control line 120 simultaneously and are started.
The programmable nature of microfluidic device provided herein be fact proved by following: the order that the change valve starts step can cause the valve in the array to be in different states, and thereby makes array be in different states.For example, in the embodiment depicted in fig. 9, can 12 samples be written into array by following steps, described step is: close separator valve 124, then control line 110 is exerted pressure, this stops using manifold 135, then reduce pressure, then reduce pressure accumulator 104 to accumulator 102.In this example, when pressure applies by accumulator 102, keep to the pressure of accumulator 104 with the control line 110 that stops valve 128 to respond to use accumulator 102 applied pressures and cause start close.On the other hand, when exerting pressure, discharge the pressure that is applied to accumulator 104 and can cause closing of valve 128 by accumulator 102.Therefore, by changing two different conditions that order that valve starts has realized microfluidic device, be written into the ability of sample or sealing input line 132 by input line 132.This programmable features provides the benefit that adopts conventional design not obtain.
Should understand concrete steps shown in Figure 9 provides a kind of concrete grammar of the microfluidic device of another embodiment of programming the present invention.Other sequence of steps also can be implemented according to some alternate embodiments.For example, alternate embodiments more of the present invention can be implemented step listed above with different orders.In addition, single step shown in Figure 9 can comprise a plurality of substeps, and it can be implemented with multiple order according to single step.In addition, can or remove other step according to concrete application interpolation.Those of ordinary skills can understand many changes, improvement and replacement.
Should understand and the invention is not restricted to concrete grammar as herein described, scheme and reagent etc., it will be understood by those skilled in the art that these variation.Will also be understood that term used herein just in order to be used to describe specific embodiment, is not intended to limit the scope of the invention.Should also be noted that as in this article with claims in used, unless spell out in addition in the context, comprise plural form when not indicating quantity.Therefore, mention one or more cell when for example, mentioning " cell ", known its equivalent of those skilled in the art.
Except as otherwise noted, all scientific and technical terminologies used herein same meaning with the technical field of the invention those of ordinary skill common sense.The details of embodiment of the present invention and various features thereof and advantage is with reference to carrying out more complete explanation at the non-limiting embodiments and the embodiment of accompanying drawing and following description details and/or explaination.Should note what feature shown in the accompanying drawing not necessarily was drawn to scale, and as understood by a person skilled in the art, even do not spell out in this article, the feature in embodiment can be used for other embodiments.Description to known assemblies and treatment technology can be omitted to avoid that embodiment of the present invention is produced unnecessary bluring.Example used herein just can effective mode for the ease of understanding the present invention, and further makes those skilled in the art implement embodiment of the present invention.Therefore, the example of this paper and embodiment should not be construed as limiting the scope of the invention, and the scope of the invention is only limited by claims and applicable law.In addition, should notice that similar Reference numeral is represented similar part in several views of accompanying drawing.
Therefore, below provided " definition " part, wherein specifically defined some terms related to the present invention, but all definition have been consistent to the understanding of these terms with those skilled in the art for clear and definite.Concrete method, equipment and material have been described, although can be used to implement or test the present invention with those any method and materials similar or that be equal to as herein described.All lists of references that this paper relates to are quoted by integral body and are incorporated this paper into.
Definition
PNA is peptide nucleic acid (peptide nucleic acid)
LNA is lock nucleic acid (locked nucleic acid)
DA is dynamic array (dynamic array)
PCR is PCR (polymerase chain reaction)
BSA is bovine serum albumin(BSA) (bovine serum albumin)
FRET is FRET (Fluorescence resonance energy transfer)
GT is Genotyping (genotyping)
PEG is polyethylene glycol (polyethylene glycol)
PLP is padlock probe (padlock probe)
Term used herein " vicinity " typically refers to the position of primer with respect to probe on the target nucleic acid analyte complementary strand.Primer and probe can be at interval about 1 to about 20 nucleotides, more specifically, about 1 to about 10 nucleotides, or can be directly adjacent one another are.
The inventive method to be used of typically referring to term used herein " analyte " detects or is quantitative with undefined nucleic acid molecules or nucleic acid molecules mixture.Term " target nucleic acid analyte " and " nucleic acid analyte " can exchange with term " analyte " in the present invention and use.
Term used herein " complementation " or " complementarity " can be included under the salt of permission and the temperature conditions polynucleotides by the natural combination of base pairing.For example, sequence " A-G-T " is in conjunction with its complementary series " T-C-A ".Complementation between two single chain molecules can be " part ", wherein has only some nucleic acid combinations, and perhaps to exist whole between single chain molecule be completely when complementary to the complementation between two single chain molecules.Complementary degree between the nucleic acid chains plays an important role to hybridization efficiency between the nucleic acid chains and intensity.This is even more important in the amplified reaction of the design and use of depending on combination between nucleic acid chains and molecule.
Term used herein " covalently bound " typically refers to a molecular structure being connected by covalent chemical bond and another molecular structure.
Term used herein " dyestuff " typically refers to any organic or inorganic molecule that absorbs electromagnetic radiation at wavelength during more than or equal to 340nm.
Term used herein " fluorescent dye " typically refers to and is being sent more any dyestuff of long wavelength's electromagnetic radiation by electromagnetic radiation source (for example, lamp, photodiode or laser) irradiation back by fluorescence mechanism.
Term used herein " GT sample buffer " typically refers to the buffer solution that the binding site of reaction channel and chamber surface in the DA chip can be sealed.Described buffer solution prevents the loss of reactive component when chip is written into processing or reaction.For reagent cost, it also can reduce the use of other Taq-Gold polymerase less than about 80%.20 * GT buffer solution can comprise following combination: betaine (FW 117.15), BSA, Superblock
Figure BPA00001406118800291
T20 (in PBS) (Thermo Scientific, Rockford, IL), Superblock (in PBS) (Thermo Scientific, Rockford, IL), Superblock
Figure BPA00001406118800293
(in TBS) (Thermo Scientific, Rockford, IL), Superblock
Figure BPA00001406118800294
T20 (in TBS) (Thermo Scientific, Rockford, IL), glycerine, PEG 20,000, PEG MME550, PEG MME5000 and Tween 20.
Term used herein " homogeneity is measured (homogenous assay) " typically refers to the method for detection or quantitative nucleic acid analyte, and it need not to measure post processing and writes down measurement result.Homogeneity is measured and can be carried out in the pipe of sealing or microfluidic arrays, and does not need to add further reagent or additional chemicals after initial with the record result measuring.Homogeneity is measured the result that the energy real time record is measured, and this means that measurement result can be recorded when making progress in time continuously in mensuration.
Term used herein " hydrolysis probes " is in U.S. Patent No. 5,210, carried out general description in 015, and it is quoted by integral body and incorporates this paper into.Hydrolysis probes has been utilized the 5 '-nuclease (TaqMan that exists in the heat endurance Taq polymerase used in the PCR reaction
Figure BPA00001406118800295
Probe technique, Applied Biosystems, Foster City CA).With fluoroscopic examination dyestuff (as fluorescein) and reception dyestuff or quencher mark hydrolysis probes.Usually, the covalently bound 5 ' end of fluorescent dye to probe, quencher is connected to 3 ' group of probe, and when probe was complete, the fluorescence that detects dyestuff was owing to FRET (FRET) and by cancellation.Probe can be in the annealing of the downstream of a primer, and described primer limits the end that PCR reaction nucleic acid target is analyzed thing amplification target position.Utilize the polymerase activity of Taq enzyme, the amplification of target nucleic acid analyte is guided with second primer that is positioned at the probe downstream but anneals on the relative chain of target nucleic acid by a primer of probe upstream.Along with the extension of upstream primer, the Taq polymerase arrives the zone of label probe annealing, and probe-template crossbred is identified as substrate, and the phosphodiester bond of hydrolysis probes.Hydrolysis irreversibly discharges the cancellation effect of quencher dyes to reporting dyes, thereby causes along with each continuous PCR circulation, and detector fluorescence increases.Particularly, hydrolysis probes of the present invention can be transcribed 8-mer common in the group or 9-mer motif and can have about 70 ℃ high T by adopting the LNA analog people and other m
Term used herein " mark " is meant to can be used for providing and can detects and/or the quantitatively any atom or the molecule of signal.Particularly, described mark can be connected to nucleic acid or protein.Mark can provide and can pass through the signal that fluorescence, radioactivity, colorimetric, X-ray diffraction or absorption, magnetic, enzymatic activity etc. detect.
Term used herein " nucleic acid " typically refers to cDNA, DNA, RNA, its strand or double-stranded and any chemical modification, as PNA and LNA.LNA is in U.S. Patent No. 6,794, describes in 499,6,670,461,6,262,490 and 6,770,748, and incorporates this paper into by quoting in full.Nucleic acid can be any size.Nucleic acid is modified can comprise the interpolation chemical group, and it is to independent nucleic acid base or extra electric charge, polarity, hydrogen bond, electrostatic interaction and the function of the whole interpolation of nucleic acid.The base that this modification can comprise modification is sugar-modified as 2 '-position, 5-position pyrimidine is modified, 8-position purine is modified, replacement, the backbone modification of the modification of cytimidine exocyclic amine, 5-bromouracil, methylate, improper base pairing combination as different cytidine of different base and different guanidine etc.Nucleic acid can come from complete chemical synthesis process (as the chemical synthesis of solid phase mediation) or come from biogenetic derivation (as by separating) or come from the process (for example dna replication dna, pcr amplification, reverse transcription) that relates to by biology tool operation nucleic acid or from the combination of these processes from almost any species that nucleic acid can be provided.
Term used herein " nucleic acid probe " is the nucleic acid that carries at least one covalently bound dyestuff (as fluorescent dye).Particularly, probe does not comprise and is used to cause the sequence of the sequence complementation of PCR reaction.
Term used herein " padlock probe " or " PLP " typically refer to the linear oligonucleotide with about 100 base-pair length.The sequence of PLP 3 ' and 5 ' end and the contiguous sequence complementation in the target nucleic acid analyte.Central incomplementarity zone at PLP has " sequence label " that can be used for discerning specific PLP.The flank of described sequence label can be universal primer site or uniqueness and/or Auele Specific Primer site, and it allows the described sequence label of pcr amplification.Hybridizing to target, 5 ' of PLP closely faces mutually with 3 ' end and can be connected subsequently.Products therefrom is the cycling probe molecule that connects the target nucleic acid analyte.For example can use TAQMAN
Figure BPA00001406118800301
PCR in real time is to label area amplification and the quantitative and/or detection of cyclisation PLP.The existence of amplicon and amount can interrelate with the existence and the amount of target sequence in the sample.To the description of PLP referring to for example Landegren etc., 2003, Padlock and proximity probes in situ and array-based analyses:tools for the post-genomic era, Comparative and Functional Genomics 4:525-30; Nilsson etc., 2006, Analyzing genes using closing and replicating circles Trends Biotechnol.24:83-8; Nilsson etc., 1994, Padlock probes:circularizing oligonucleotides for localized DNA detection, Science 265:2085-8.Above-mentioned list of references is incorporated this paper into by quoting in full.
Term used herein " PCR " typically refer to be used to increase, the method for detection or quantifying analytes specific region.Those skilled in the art will appreciate that on the basic PC R technical foundation several variations are arranged, as allele specific pcr, set PCR (assembly PCR) or polymerase cycle set (polymerase cycling assembly, PCA), colony PCR, helicase dependent amplification, heat start PCR, specificity (intersequence-specific between sequence, ISSR) PCR, inverse PCR, ligation-mediated PCR, methylation status of PTEN promoter, multidigit point join dependency probe amplification, multidigit point PCR, nest-type PRC, overlapping extension PCR, quantitative PCR, quantitative PCR in real time, RT-PCR, hot asymmetric interlaced (thermal asymmetric interlaces, TAIL) PCR, touchdown PCR and PAN-AC.In addition, how to it will be understood by those skilled in the art that these change application on the round pcr on basis.
Term used herein " purifying " typically refers to according to electric charge, molecular size or the binding affinity any process with protein, polypeptide or nucleic acid and other compositions or compound separation.
Term used herein " quencher " typically refers to the dyestuff of the fluorescent emission that reduces another dyestuff.
Term used herein " inquiry (querying) " typically refers to determines target specific probe whether relevant with nucleic acid analyte (for example, connecting or link), and the amount of target specific probe in the randomly quantitative sample.
" sample " used herein typically refers to tissue or the fluid sample from the human or animal, include but not limited to blood plasma, serum, spinal fluid, lymph liquid, the outer section of skin, respiratory system, intestines and urogenital tract, tears, saliva, haemocyte, tumour, organ, tissue and cell in vitro are cultivated the sample of component.Particularly, sample can be unicellular, paraffin-embedded tissue sample and the biological biopsy samples of acupuncture.In addition, sample can comprise environmental sample, as lake water and foodstuff samples.
Speech used herein " basic purifying " or " basic separation " generally include nucleic acid or the amino acid sequence obtaining, separate or separate from their natural surroundings, they do not contain at least about 60%, especially at least about 75%, the most especially at least about other components that are associated with it of 90%, and comprise reorganization or clone's separate nucleic acid thing and chemical synthesis analog or the synthetic analog of system biological.
Consider the very big diversity of polymer chemistry character, precursor, synthetic method, reaction condition and available additive, have a large amount of useful elastomers systems to can be used for making elastic module, layer, film, miniature valve, pump etc.The variation of material therefor in some cases can be owing to the specific material behavior of needs, and promptly solvent resistance, rigidity, gas permeability or temperature stability decide.The elastomeric polymer that a lot of types are arranged.This paper provides the simple description of modal elastomer type, even purpose is to show to adopt the polymer of " standard " relatively, also has the multiple possibility of combination.Elastomeric polymer commonly used comprises polyisoprene, polybutadiene, polychlorobutadiene, polyisobutene, poly-(s-B-S), polyurethane and silicone or polysiloxanes.
Polyisoprene, polybutadiene and polychlorobutadiene are polymerized by diene monomers, and therefore each monomer has a two key when polymerization.This pair key makes polymer pass through sulfuration (usually, sulphur is used for forming crosslinked by heating) between two keys and is converted into elastomer.This makes and is easy to the soft printing of homogeneous multilayer (homogeneous multilayer soft lithography) by the incomplete vulcanization for the treatment of binder course; Can carry out photoresist encapsulation (photoresist encapsulation) by similar mechanism.
Pure polyisobutene does not have two keys, but by introduce when the polymerization a small amount of (about 1%) thus isoprene carry out crosslinked as elastomer.This isoprene monomer provides two keys that dangle on the polyisobutene skeleton, it can cure subsequently as described above.
Poly-(s-B-S) produces by active anionic polymerization (i.e. no natural chain termination step in reaction), therefore can have " activity " polymer ends in the cured polymer kind.This is the natural material standed for (wherein having a large amount of unreacted monomers in the liquid layer that is cast in cured layer top) of photoresist package system.Incomplete curing makes can spare the soft printing of matter multilayer (A combines with A).This chemical property also is beneficial to and makes one deck have extra butadiene (" A ") and coupling agent, and another layer (" B ") lacks butadiene (being used for the even soft printing of matter multilayer).SBS is " thermo-setting elastomer ", means its fusing and plastic (relative with elasticity) on uniform temperature; Reduce temperature and produce elastomer again.Therefore, layer can combine by heating.
Polyurethane is made by diisocyanate (A--A) and glycol or diamines (B--B); Because diisocyanate and glycol/amine have the enormous amount of a variety of, dissimilar polyurethane.Yet the A of polymer makes them be used for the soft printing of heterogeneous multilayer as RTV 615 with respect to the characteristic of B: by using excessive A-A at one deck, use excessive B-B in another layer.
Siloxane polymer has great structural change, and multiple commercial preparation is provided.The vinyl of RTV 615-to (Si--H) crosslinked (can carry out the encapsulation of soft printing of heterogeneous multilayer and photoresist) has been discussed, but this just is used for one of several cross-linking methods of siloxane polymer chemistry.
Except adopting above-mentioned simple " pure " polymer, can add crosslinking agent.Some reagent monomer of the two keys that dangle that are used to vulcanize (as have) are suitable for soft printing of homogeneous (A to A) multilayer or photoresist encapsulation; In two elastic layers, mix identical reagent in the method.Complementary reagent (be that a kind of monomer has two keys that dangle, another kind has the Si-H of dangling group) is suitable for the soft printing of heterogeneous (A to B) multilayer.Add complementary agent to adjacent layer in the method.
Below for uniting the elastomeric non-exhaustive list of use: polyisoprene, polybutadiene, polychlorobutadiene, polyisobutene, poly-(s-B-S), polyurethane and siloxane polymer with the present invention; Or poly-(two (fluoroalkyl) phosphine nitrile) (PNF, Eypel-F), poly-(carborane-siloxanes) (Dexsil), poly-(acrylonitrile-butadiene) (acrylonitrile-butadiene rubber), poly-(1-butylene), poly-(chloro trifluoro-ethylene-difluoroethylene) copolymer (KeI-F), poly-(ethyl vinyl ether), poly-(difluoroethylene), poly-(difluoroethylene-hexafluoropropene) copolymer (Viton), polyvinyl chloride elastic composition (PVC), polysulfones, Merlon, polymethyl methacrylate (PMMA) and polytetrafluoroethylene (PTFE) (Teflon).
Allcock etc., Contemporary Polymer Chemistry, second edition is described as being present in polymer under the temperature between glass transition temperature and the condensing temperature with elastomer.The elastomer display elastic characteristic be because, the polymer chain motion that is easy to twist is untied skeletal chain under the effect of power, curl and return originally shape in the unable time spent skeletal chain of doing.Usually, the elastomer distortion reverts to original shape when removing power when applying power.The elasticity of elastomer display can characterize with modulus of elasticity.The modulus of elasticity of the material that embodiments more of the present invention are used for about 1Pa to about 1TPa or about 10Pa to about 100GPa or about 20Pa about 1GPa or about 50Pa about 10MPa or about 100Pa about 1MPa extremely extremely extremely, although also can adopt these extraneous moduluss of elasticity according to the needs of concrete application.In some cases, the modulus of elasticity of material can be about 100MPA (MPa) or still less.In other embodiments, the modulus of elasticity of material for about 75MPA or still less, about 50MPA or still less, about 25MPA or still less, about 10MPA or still less, about 8MPA or still less, about 5MPA or still less or about 2MPA or still less.
Embodiment of the present invention provide microfluidic device, and it comprises assembly for example passage, valve and chamber, described assembly to small part by fast one or more of springform layer or level comprises, embedding, perhaps form or form therein by it.A kind of exemplary microfluidic device has reagent circulation road or the reagent line that is formed in the elastomer ground floor.Described reagent circulation road comprises and holds valve and chamber conduit.Microfluidic device also can have be formed at the adjoining elastomer second layer of described ground floor in control channel or hold line.In addition, microfluidic device can comprise be formed at the 3rd layer on the adjoining elastomer of the described second layer in sample flow passage or sample wire.Described sample flow passage can comprise and hold valve and chamber conduit.Described control channel can be held valve with the reagent circulation road and hold the valve operability with the sample flow passage and be connected.Described microfluidic device can comprise reagent chamber that is communicated with reagent linear flow body and the sample room that is communicated with the sample wire fluid.Described reagent chamber and sample room can be by being formed at reaction stream passage or response line and the fluid communication with each other in the 3rd layer on the described elastomer.Described response line can comprise interface valve.Described microfluidic device also can comprise be formed at the 4th layer on described the 3rd layer of adjoining elastomer in interface channel.Described interface channel can be connected with described reaction stream Entry Interface valve operability.
Embodiment of the present invention also contain the method for making and using microfluidic device disclosed herein.For example, the operation of microfluidic device can comprise opens one or more separate valves, closes one or more interface valve and make material flow through separate valves and flow into one or more chamber, randomly carries out under pressure.Technology also can comprise the pressure that changes in the potted line to close separate valves, and with the sealing separate chambers, and the pressure of change parting line is to open interface valve.The interface valve that first material of first Room can flow through unlatching enters second Room, and wherein first material mixes with wherein contained second material or reacts.
Should understand example as herein described and embodiment just for the example purpose, those skilled in the art can advise multiple improvement or change according to it, and these are included in the application's spirit and the authority and in the scope of claims.

Claims (120)

1. microfluidic device, it comprises
Pressure source;
Control line, it is communicated with described pressure source fluid;
A plurality of valves, it is by described control line operation; With
Separate valves, it is near the described control line and be between described pressure source and the described a plurality of valve.
2. the microfluidic device of claim 1, wherein said separate valves make up and are arranged in and block fluid flowing by described control line.
3. the microfluidic device of claim 1, wherein said separate valves is communicated with the second pressure source fluid.
4. the microfluidic device of claim 1, wherein said control line comprises the locking control line that comprises check valve.
5. the microfluidic device of claim 4, wherein said check valve are set to stop fluid to flow to described pressure source from described valve.
6. the microfluidic device of claim 5, it also comprises a plurality of chambers valve.
7. the microfluidic device of claim 6, wherein said control line provide to described a plurality of valves and described a plurality of chambers valve and start pressure.
8. the microfluidic device of claim 1, wherein said separate valves can be operated and is used to stop described a plurality of valve to close.
9. the microfluidic device of claim 1, it also comprises more than second valve by the operation of second control line.
10. the microfluidic device of claim 9, wherein said second control line comprises the second locking control line that comprises second check valve.
11. operation has the method for the microfluidic device of valve and control line, described control line has the valve group that is associated with it, and described method comprises:
Close described valve; With
Described control line is exerted pressure; Wherein the valve of cutting out can not be operated the described valve group that is associated with described control line.
12. the method for claim 11 is wherein closed described valve and is comprised:
Second control line is applied second pressure; With
Respond to described second pressure and close described valve.
13. the method for claim 11, wherein the first valve group responds to described first control line is applied first pressure and closes.
14. the method for claim 13, the wherein said first valve group be by locking,
15. the method for claim 11, it also comprises:
Keep being applied to described second pressure of described second control line; With
Described first control line is applied second pressure less than described first pressure, and wherein the valve group that is associated with described second control line responds to described second the applying of pressure and closes.
16. the method for claim 15, wherein said valve group is by locking.
17. a microfluidic device, it comprises:
First valve;
Second valve;
Control line, it is communicated with described first valve and the described second valve fluid;
The acute build up of pressure device, it is communicated with described control line fluid; With
Check valve, it is near the described control line and between described acute build up of pressure device and described second valve.
18. the microfluidic device of claim 17, the described control line between wherein said acute build up of pressure device and described first valve does not have check valve.
19. the microfluidic device of claim 17, wherein said first valve is set to the reative cell that is arranged in the described microfluidic device is separated.
20. the microfluidic device of claim 17, wherein said second valve are set to that first fluid is imported line and second fluid input line is separated.
21. the microfluidic device of claim 17, it also comprises the 3rd valve that is communicated with described control line fluid.
22. the microfluidic device of claim 21, wherein said the 3rd valve can operate be used to stop fluid by with the flowing of the mutually coupled fluid input line of reative cell.
23. the microfluidic device of claim 21, it also comprises structure and is arranged to the separator valve that the obstruction fluid flows through described control line, and wherein said separator valve places between described acute build up of pressure device and described the 3rd valve along described control line.
24. the microfluidic device of claim 23, wherein said separator valve are set to accept the pressure that starts from second control line, described second control line is communicated with the second acute build up of pressure device fluid.
25. the microfluidic device of claim 24, wherein said second control line is independent of described control line.
26. the microfluidic device of claim 23, wherein said separator valve can be operated and be used to stop described the 3rd valve to respond to starting of described control line and close.
27. a microfluidic device, it comprises:
A plurality of reative cells, it is arranged as array layout, second valve that each of described a plurality of reative cells has first valve that is communicated with one of described a plurality of reative cells fluid and be communicated with one of the described fluid of described a plurality of reative cells;
First control line, it can be operated in order to start described first valve and described second valve;
Input line group, it is communicated with described a plurality of reative cell fluids;
A plurality of sample inlets, it is communicated with described input line group fluid; With
Check valve, it is arranged in described first control line.
28. the microfluidic device of claim 27, it also comprises and is communicated with the first acute build up of pressure device fluid and can be operated in order to block the first valve group of a part of described input line group.
29. the microfluidic device of claim 27, the wherein said first valve group is in the downstream of described check valve.
30. the microfluidic device of claim 27, it also comprises:
The second input line group, it is communicated with described a plurality of reative cell fluids; With
More than second sample inlet, it is communicated with the described second input line group fluid.
31. the microfluidic device of claim 30, it also comprises can be by the second valve group of second control line operation.
32. the microfluidic device of claim 31, wherein said second control line is communicated with the second acute build up of pressure device fluid, and can be operated in order to block the described second input line group of a part.
33. the microfluidic device of claim 31, it also comprises second check valve that is arranged in described second control line.
34. the microfluidic device of claim 31, it also comprises and is arranged in described second control line and can be operated in order to block the separator valve that fluid flows through described first control line.
35. a microfluidic device, it comprises:
A plurality of reative cells;
A plurality of first input ports, each of wherein said a plurality of first input ports is communicated with one or more fluids in described a plurality of reative cells by one of more than first input lines;
A plurality of second input ports, each of wherein said a plurality of second input ports is communicated with one or more fluids in described a plurality of reative cells by one of more than second input lines;
The first acute build up of pressure device, it is communicated with the first control line fluid, and wherein said first control line is set to close described more than first input lines;
The second acute build up of pressure device, it is communicated with the second control line fluid, and wherein said second control line is set to close described more than second input lines;
First check valve, it is arranged in described first acute build up of pressure device and described more than first individual described first control lines of importing between the lines; With
Second check valve, it is arranged in described second acute build up of pressure device and described more than second individual described second control lines of importing between the lines.
36. the microfluidic device of claim 35, it also comprises and can be operated in order to separate each valve group of described a plurality of reative cell.
37. the microfluidic device of claim 35, it also comprises and can be operated in order to block described second control line and to be arranged in valve between described second acute build up of pressure device and described second control line.
38. the microfluidic device of claim 37, wherein said first control line can be operated and be used to start described valve.
39. the microfluidic device of claim 35, it also comprises the valve groups that can operate the subgroup that is used to block described more than second input lines.
40. the microfluidic device of claim 35, it also comprises can operate the valve group that is used to block manifold, and described manifold provides described more than second fluids between the input line to be communicated with.
41. operation has the method for the microfluidic device of a plurality of valves and check valve, described method comprises:
Control line to described microfluidic device applies first fluid pressure;
Respond to described first the applying of pressure and close described a plurality of valve;
Respond to described first the applying of pressure and close described check valve; With
Second control line to described microfluidic device applies second fluid pressure.
42. the method for claim 41, it also comprises and reduces the described first fluid pressure be applied in the described control line.
43. the method for claim 42, wherein after reduction was applied to described first fluid pressure in the described control line, described a plurality of valves kept closed conditions.
44. the method for claim 42 wherein reduces described first fluid pressure and comprises described control line is not applied fluid pressure.
45. the method for claim 41 wherein applies described second fluid pressure to described second control line and comprises and close separator valve.
46. can operating the fluid that is used to stop between described a plurality of valve and the described control line, the method for claim 45, wherein said separator valve be communicated with.
47. the method for claim 41, wherein said microfluidic device have a plurality of reative cells and with described a plurality of reative cells in each valve group that is associated.
48. operation has the method for the microfluidic device of a plurality of input ports, described method comprises:
Provide the input fluid to one of described a plurality of input ports;
Start the valve group closing the first of the input line that is connected with the subgroup of described a plurality of input ports, wherein said subgroup does not comprise the one of described of described a plurality of input ports;
Make described input fluid flow through one of the described input line that links to each other with described a plurality of input ports;
Make described input fluid through the second portion of described input linear flow to described input line; With
Close the second valve group to separate a plurality of reative cells.
49. the method for claim 48 is wherein closed the described second valve group and is comprised first pressure source is applied fluid pressure.
50. the method for claim 49 is wherein closed the described second valve group and is comprised closing the manifold that fluid is communicated with between the input line is provided.
51. the method for claim 49 is wherein started described valve group and is comprised second pressure source is applied fluid pressure.
52. the method for claim 48, the described second portion that wherein makes described input fluid flow to described input line comprise that described input fluid is flow through provides the manifold that fluid is communicated with between the described input line.
53. the method for claim 48 was wherein started described valve group before closing the described second valve group.
54. the method for claim 48, it also comprises makes described input fluid flow through the second input line group, and the described second input line group of branches becomes in the described input line the described second portion of each.
55. the method for operation programmable microfluidic device, described microfluidic device have the reaction site array that is communicated with the first input line group and the second input line group fluid, described method comprises:
Start and to be operated in order to block the first valve group of the described first input line group;
Start the second valve group that can be operated in order to the first of the subgroup of blocking the second input line group;
Second portion by the described second input line group is loaded into described reaction site with sample; With
Start and to be operated in order to separate the 3rd valve group of described reaction site.
56. the method for claim 55 is wherein started the described first valve group and is comprised the described first valve group of locking.
57. the method for claim 55 is wherein started the described first valve group and is comprised to first pressure source that is communicated with the described first valve group fluid and apply first pressure.
58. the method for claim 55 is wherein started the described first valve group and is started the described second valve group and carries out simultaneously.
59. the method for claim 55 is wherein started described the 3rd valve group and is comprised to second pressure source that is communicated with described the 3rd valve group fluid and apply second pressure.
60. the method for claim 59, starting after a plurality of samples are loaded into described reaction site of wherein said the 3rd valve group carried out.
61. the method for claim 55, wherein said sample flow to the second portion of the described second input line group through the manifold of the second portion that connects the described second input line group from the first of described input line, and the first of described input line is not included in the subgroup of the described second input line group.
62. the method for claim 55, the input line that wherein is not included in the described subgroup of the described second input line group is communicated with the input port fluid that is set to receive described sample.
63. a microfluidic device, it comprises:
The input port of predetermined quantity, each described input port can be operated in order to receive one of a plurality of input fluids;
A plurality of input fluid lines, each of described a plurality of input fluid lines is communicated with one of the input port of described predetermined quantity fluid;
Valve group, each of described valve group can be operated in order to close one of described a plurality of input fluid lines, and the quantity of wherein said valve group is less than described predetermined quantity;
Manifold, it is communicated with each fluid of described a plurality of input fluid lines; With
The second valve group, each of the described second valve group can be operated in order to close the part of described manifold.
64. the microfluidic device of claim 63, wherein said predetermined quantity are 12.
65. the microfluidic device of claim 63, it also comprises first pressure source that is communicated with described valve group fluid.
66. the microfluidic device of claim 65, it also comprises second pressure source that is communicated with described valve group fluid.
67. the microfluidic device of claim 63, the described part of wherein said manifold comprises fluid line, and described fluid line links to each other with another of described a plurality of input fluid lines one of described a plurality of input fluid lines.
68. the microfluidic device of claim 63, the quantity of wherein said valve group are the quantity less than described predetermined quantity.
69. the microfluidic device of claim 63, each of wherein said a plurality of input fluid lines are branched off into a plurality of input fluid lines.
70. the method for operation programmable microfluidic device, described microfluidic device has the reaction site array, and it is communicated with the manifold fluid that is connected the described second input line group with the first input line group, the second input line group, and described method comprises:
Start and to be operated in order to close the first valve group of the described first input line group;
Start the second valve group that can be operated in order to the first of the subgroup of closing the second input line group;
Start to be operated and use so that the 3rd valve group that described manifold is stopped using;
Cancel and start the described second valve group;
Second portion by the described second input line group is loaded into a plurality of samples in the described reaction site; With
Start and to be operated in order to separate the 4th valve group of described reaction site.
71. the method for claim 70 is wherein started the described first valve group and is comprised the described first valve group of locking.
72. the method for claim 70 is wherein started the described first valve group and is started the described second valve group and comprises first pressure source is applied first pressure that described first pressure source is communicated with described first valve group and the described second valve group fluid.
73. the method for claim 72 is wherein started described the 3rd valve group and is comprised second pressure source is applied second pressure that described second pressure source is communicated with the described second valve group fluid.
74. the method for claim 73, wherein said second pressure is less than described first pressure.
75. the method for claim 70 is wherein cancelled and is started the described second valve group and comprise described first pressure that reduces to be applied to described first pressure source.
76. the method for claim 70 is wherein started described the 3rd valve group and is comprised described the 3rd valve group of locking.
77. the method for claim 70, it comprises also and starts valve stoping closing of the 5th valve group that described the 5th valve group can be operated in order to close the described second portion of described second group of input line.
78. the method for claim 77, wherein said valve is communicated with the first pressure source fluid, and described first pressure source is communicated with the described first valve group fluid.
79. the method for claim 70, the first of each is communicated with the input port fluid in the wherein said second input line group, and described input port is set to receive one of described a plurality of samples.
80. the method for claim 70, wherein said reaction site comprises a plurality of reative cells, and each of described a plurality of reative cells is communicated with by the second portion fluid of through hole with one of described second input fluid line group.
81. the method for claim 70, the first of each is communicated with different input ports fluid in the wherein said second input line group.
82. the method for operation programmable microfluidic device, described programmable microfluidic device have the reaction site array that is communicated with the first input line group and the second input line group fluid, described method comprises:
Start and to be operated in order to block the first valve group of the described first input line group;
Start and to be operated in order to separate the second valve group of described reaction site;
Cancel and start the described second valve group;
A plurality of samples are written into described reaction site by the second input line assembling; With
Start the described second valve group.
83. the method for claim 82 is wherein started the described first valve group and is comprised the described first valve group of locking.
84. the method for claim 82 is wherein started the described first valve group and is comprised first pressure source is applied first pressure that described first pressure source is communicated with the described first valve group fluid.
85. the method for claim 82, it also is included in and starts the 3rd valve group when starting the described first valve group.
86. the method for claim 85 is wherein started described the 3rd valve group and is comprised described the 3rd valve group of locking.
87. the method for claim 82 is wherein cancelled and is started the described first valve group and comprise described first pressure that reduces to be applied to described first pressure source.
88. the method for claim 82, each is communicated with different input ports fluid in the wherein said second input line group.
89. the method for claim 82 is wherein started the described first valve group and is started the described second valve group and carries out simultaneously.
90. the method for claim 82, wherein said reaction site comprises the reative cell in the elastomer layer that is limited to described microfluidic device.
91. a microfluidic device, it comprises:
A plurality of reaction site;
The first input line group, it provides first input port of predetermined quantity to be communicated with fluid between described a plurality of reaction site, and wherein said first group quantity is described predetermined quantity;
The second input line group, it provides second input port of predetermined quantity to be communicated with fluid between described a plurality of reative cells, and wherein said second imports that each comprises trunk portion and component in the line group, and described second group quantity is less than described predetermined quantity; With
Input unit able to programme, it can be operated in order to use described first input line group or the described second input line group to fill described reative cell.
92. the microfluidic device of claim 91, wherein said a plurality of reaction site are arranged as array layout.
93. the microfluidic device of claim 91, each comprises reative cell in wherein said a plurality of reaction site, and described reative cell is communicated with one of described first input fluid line group fluid by through hole.
94. the microfluidic device of claim 91, each comprises reative cell in wherein said a plurality of reaction site, and described reative cell is imported one of the component of fluid line group fluid and is communicated with by through hole and described second.
95. the microfluidic device of claim 91, the described component of each forms four fluid passages from described trunk portion in the wherein said input line group.
96. comprising, the microfluidic device of claim 91, wherein said input unit able to programme can be operated in order to close the valve group of the described first input fluid line group.
97. the microfluidic device of claim 96, wherein said valve group is communicated with the check valve fluid, and described check valve can be operated in order to described valve group is latched in closed condition.
98. the microfluidic device of claim 91, wherein said input unit able to programme comprises the valve group, and described valve group can be operated in order to close the described component of the described second input fluid line group.
99. the microfluidic device of claim 98, wherein said valve group is communicated with the check valve fluid, and described check valve can be operated in order to described valve group is latched in closed condition.
100. the microfluidic device of claim 98, it also comprises and is arranged near the described valve group and can be operated separate valves in order to stop described valve group to be closed.
101. the method for the microfluidic device with a plurality of control lines is set, and described method comprises:
Start first control line;
Valve is placed first state;
Afterwards, start and can be operated in order to the valve group is placed second control line of second state, the valve that wherein is in described first state stops described valve group to be placed in described second state.
102. the method for claim 101, wherein said first state is to close.
103. the method for claim 101, wherein said second state is to close.
104. the method for claim 101 is wherein started described first control line and is comprised first pressure source is applied first pressure.
105. the method for claim 104 is wherein started described second control line and is comprised second pressure source is applied second pressure.
106. the method for claim 105, wherein said second pressure source is independent of described first pressure source.
107. a setting has the method for the microfluidic device of a plurality of control lines, described method comprises:
Then start first state that second control line is set up described microfluidic device by starting first control line; With
Then start described first control line and set up described second state of fluid means that is by starting described second control line.
108. the method for claim 107, wherein said first state comprise the first input line group that is in closed condition and is communicated with a plurality of reative cell fluids of being arranged on the described microfluidic device second import the line group.
109. the described second input line group that the method for claim 108, wherein said second state comprise the described first input line group that is communicated with described a plurality of reative cell fluids of being arranged on the described microfluidic device and be in closed condition.
110. the method for claim 108, wherein said second state also comprises the valve group that is in closed condition, and it stops fluid to flow through the manifold of the described second input line group of connection.
111. the method for claim 107, wherein said first control line is communicated with the first pressure source fluid.
112. the method for claim 111, wherein said second control line is communicated with the second pressure source fluid.
113. the method for claim 107, wherein said microfluidic device comprises elastomeric material.
114. a microfluid system, it comprises:
Carrier, it comprises:
A plurality of first input ports;
A plurality of first input lines, each is communicated with one of described a plurality of first input ports fluid in described a plurality of first input lines;
A plurality of second input ports;
A plurality of second input lines, each is communicated with one of described a plurality of second input ports fluid in described a plurality of second input lines;
First pressure source; With
Second pressure source
Be contained in the microfluidic device on the described carrier, described microfluidic device comprises:
A plurality of the 3rd input lines, each is communicated with one of described a plurality of first input lines fluid in described a plurality of the 3rd input lines;
A plurality of the 4th input lines, each is communicated with one of described a plurality of second input lines fluid in described a plurality of the 4th input lines;
First control line that is communicated with the described first pressure source fluid;
Check valve, it can be operated in order to block at least a portion of described first control line; With
Second control line that is communicated with the described second pressure source fluid.
115. the microfluid system of claim 114, wherein said a plurality of first input ports can be operated in order to receive the UP source.
116. the microfluid system of claim 114, wherein said a plurality of second input ports can be operated in order to receive the UP source.
117. the microfluid system of claim 114, it also comprises and can be operated in order to block second check valve of described second control line at least a portion.
118. the microfluid system of claim 114, wherein said microfluidic device also comprises manifold, and it is connected the described a plurality of second input lines in the described carrier with a plurality of the 4th input lines described in the described microfluidic device.
119. the microfluid system of claim 118, it also comprises the valve group that can be operated in order to the part of blocking described manifold.
120. the microfluid system of claim 114, wherein said microfluidic device also comprise a plurality of reative cell groups, each reative cell of each reative cell group is communicated with one of described a plurality of the 3rd input lines and one of described a plurality of the 4th input lines fluid.
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