CN105964316A

CN105964316A - Programmable microfluidic digital array

Info

Publication number: CN105964316A
Application number: CN201610459013.4A
Authority: CN
Inventors: 布赖恩·福勒
Original assignee: Fluidigm Corp
Current assignee: Standard Biotools Corp
Priority date: 2008-12-08
Filing date: 2009-12-07
Publication date: 2016-09-28
Also published as: SG10201404682WA; EP2376227A4; EP2376227A1; CN104741159A; CN102281950A; SG10201404685PA; CN102281950B; WO2010077618A1; SG172015A1; SG10201404683TA; CN104741159B

Abstract

The invention relates to a microfluidic device, and more specifically relates to a programmable microfluidic digital array; the 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

Programmable microfluidic digital array

The application is the divisional application of the Chinese patent application of Application No. 201510158081.2, former Application is the divisional application of the Chinese patent application of application number 200980154761.X, original application 200980154761.X be the PCT international application of December in 2009 submission on the 07th PCT/US2009/067037 enters the application of National Phase in China on July 15th, 2011.

Technical field

The present invention relates to Programmable microfluidic digital array.

Background technology

Microfluidic device can be for analyzing, prepare, measuring on the yardstick the most still cannot imagined And other operating functions.The advantage of microfluidic device includes saving precious reagent and sample, sample Analyze or synthesis high density and high flux, the most visible level of naked eyes on fluid degree of accuracy and Accuracy and the replacement space that the corresponding device of operation is brought on macrofluid yardstick are reduced.With Microfluidic device size reduces and density increase relevant be higher complexity and bigger engineering with Manufacturing cost and finer device structure.

Although having these to be in progress in microfluid design and application aspect, but reduce answering of micro-fluid chip Miscellaneous degree and simplify they operation or useful.Furthermore, it is necessary to strengthen control fluid flowing with And the ability of the correlated response process occurred in microfluidic device.Therefore, this area needs and microfluid The method and system of the improvement that device is relevant.

Summary of the invention

The present invention relates to microfluidic device.More particularly it relates to programmable microfluidic Digital Arrays Row and operational approach thereof.Being only used as citing, described method and apparatus has been applied in micro-fluid chip There is provided in the system of asynchronous logic function.Additionally, check valve is integrated by some embodiments of the present invention Enter digital array to provide lock-in control line.However, it is recognized that the present invention has much broader being suitable for Scope.

According to one embodiment of the invention, it is provided that microfluidic device.Described microfluidic device bag Containing pressure source and with pressure source fluid communication control line.Described microfluidic device also comprises multiple passing through The valve of described control line operation, also comprises separate valves, and described separate valves is positioned near described control line also Between described pressure source and the plurality of valve.

According to another embodiment of the invention, it is provided that operation has the microfluid of valve and control line The method of device, described control line has coupled valve group.Described method includes closing described valve And described control line is applied pressure.The valve of described cut out causes the valve group that is connected with described control line not Operable.

According to yet another embodiment of the present invention, it is provided that microfluidic device.Described microfluidic device Comprise the first valve, the second valve and the control line being in fluid communication with described first valve and the second valve.Described micro- Fluid means also comprises the acute build up of pressure device (pressure being in fluid communication with described control line Accumulator) the described control line and between described acute build up of pressure device and described second valve Neighbouring check valve.

An alternate embodiment according to the present invention, it is provided that microfluidic device.Described microfluid Device comprises multiple reative cell arranged with array layout.Each of the plurality of reative cell have with One of multiple reative cells fluid communication the first valve and with the plurality of reative cell in described in one of Second valve of fluid communication.Described microfluidic device also comprises operable to start described first valve and institute The first control line stating the second valve and the input line group being in fluid communication with the plurality of reative cell.Described Microfluidic device also comprises the multiple sample inlets with described input line group fluid communication and is arranged in Check valve in described first control line.

Another alternate embodiment according to the present invention, it is provided that microfluidic device.Described miniflow Body device comprises multiple reative cell and multiple first input port.The each of the plurality of first input port is led to Cross one or more fluid communication of one of more than first input line and the plurality of reative cell.Described micro- Fluid means also comprises multiple second input port.The plurality of second input port each by more than second One or more fluid communication of one of individual input line and the plurality of reative cell.Described microfluidic device Also comprise and the first acute build up of pressure device of the first control line fluid communication.Described first control line is set to Close described more than first input line.Additionally, described microfluidic device comprises and the second control line fluid Second acute build up of pressure device of connection.Described second control line is set off described more than second inputs Line.It addition, described microfluidic device comprises is arranged in described first acute build up of pressure device and described more than first The first check valve in described first control line between individual input line, and it is arranged in described second pressure The second check valve in described second control line between accumulator and described more than second input line.

According to another alternate embodiment, it is provided that operation has the micro-of multiple valve and check valve The method of fluid means.Described method includes applying first fluid to the control line of described microfluidic device Pressure, and respond to the applying of described first pressure and close the plurality of valve.Described method is also wrapped Include and respond to the applying of described first pressure and close described check valve, and to described microfluidic device Second control line apply second fluid pressure.

According to one embodiment of the invention, it is provided that operation has the miniflow of multiple input port The method of body device.Described method includes providing input fluid to one of the plurality of input port, and Start the Part I input line that valve group is connected with the subgroup of the plurality of input port with closedown.Described Asia Group does not comprise the one of described of the plurality of input port.Described method also includes making described input fluid stream Cross the input line being connected with one of described in the plurality of input port, make described input fluid flow through described Input line arrives the Part II of described input line and closes the second valve group to separate multiple reative cells.

Another specific embodiments according to the present invention, it is provided that operation programmable microfluidic device Method, described device has the reaction site being in fluid communication with the first input line group and the second input line group Array.Described method includes starting operable the first valve group in order to block described first input line group, And start the second valve group of the Part I of the operable subgroup in order to block the second input line group.Institute Method of stating also includes that sample is loaded into described reaction by the Part II by described second input line group Site and start operable the 3rd valve group in order to separate described reaction site.

Another specific embodiments according to the present invention, it is provided that microfluidic device.Described microfluid Device comprises the input port of predetermined quantity and multiple input fluid line, described input port each operable To receive one of multiple input fluids, each and described predetermined quantity of the plurality of input fluid line One of input port is in fluid communication.Described microfluidic device also comprises valve group, and described each of valve group grasps Make to close one of the plurality of input fluid line.The quantity of described valve group is less than described predetermined quantity. Described microfluidic device also comprises the manifold and second of each fluid communication with described input fluid line Valve group, described second valve group each operable with close described manifold a part.

According to one embodiment of the invention, it is provided that the method for operation programmable microfluidic device, Described device has reaction site array, its with the first input line group, the second input line group and with described The manifold fluid connection that second input line group connects.Described method includes starting operable in order to close State the first valve group of the first input line group, start the operable subgroup in order to close the second input line group Second valve group of Part I, and start and be operable to make described manifold disable (deactivate) the 3rd valve group.Described method also includes cancelling starts described in (deactuate) Multiple samples are loaded into described reaction by the Part II of described second input line group by the second valve group Site, and start the 4th valve group being operable to separate described reaction site.

According to another embodiment of the invention, it is provided that the side of operation programmable microfluidic device Method, described device has the reaction site battle array being in fluid communication with the first input line group and the second input line group Row.Described method includes starting the first valve group being operable to block described first input line group, And start the second valve group being operable to separate described reaction site.Described method also includes removing Pin starts described second valve group, is assembled by the second input line by multiple samples and is loaded into described reaction site And start described second valve group.

According to yet another embodiment of the present invention, it is provided that microfluidic device.Described microfluidic device Comprising multiple reaction site and the first input line group, described first input line group provides the of predetermined quantity Fluid communication between one input port and the plurality of reaction site.The quantity of described first group is described Predetermined quantity.Described microfluidic device also comprises the second input line group, and it provides the second of predetermined quantity Fluid communication between input port and the plurality of reative cell.Described second each of input line group comprises Trunk portion and component, and the quantity of described second group is less than described predetermined quantity.Described micro- Fluid means also comprises programmable input equipment, and it is operable in order to use described first input line group Or described second input line assemble fill out described reative cell.

According to one embodiment of the invention, it is provided that arrange the miniflow with multiple control line The method of body device.Described method includes starting the first control line and valve being placed in the first state.Institute Method of stating also includes starting the second control line being operable to that valve group is placed in the second state afterwards. The valve being in described first state stops described valve group to be placed in the second state.

Another specific embodiments according to the present invention, it is provided that arrange and there is the micro-of multiple control line The method of fluid means.Described method includes by starting the first control line and then starting the second control The first state of described microfluidic device set up by line, and by start described second control line and with After start described first control line and set up the second state of described microfluidic device.

Another specific embodiments according to the present invention, it is provided that a kind of microfluidic device.Described micro- Fluid means comprises the first valve and the second valve.Described microfluidic device also comprises and described first valve and institute State the control line of the second valve fluid communication, and the pressure source being in fluid communication with described control line.Described Microfluidic device also comprises and is arranged in pressure source described in described control line and between described second valve Check valve.

Another alternate embodiment according to the present invention, it is provided that microfluid system.Described miniflow System system comprises carrier.Described carrier comprises multiple first input port and multiple first input line.Described One of each and the plurality of first input port of multiple first input lines is in fluid communication.Described carrier is also Comprise multiple second input port and multiple second input line.Each and the institute of the plurality of second input line State one of multiple second input port fluid communication.Described carrier also comprises the first pressure source and the second pressure Source.Described microfluid system also comprises the microfluidic device being contained on described carrier.Described microfluid Device comprises multiple 3rd input line and multiple 4th input line.The plurality of 3rd input line each It is in fluid communication with one of the plurality of first input line, described 4th input line each and the plurality of One of second input line is in fluid communication.Described microfluidic device also comprises and described first pressure source fluid First control line of connection, is operable to block described first control line at least one of unidirectional Valve, and the second control line being in fluid communication with described second pressure source.

The present invention has the many advantages surmounting routine techniques.Such as, the technology of the present invention allows manufacturing Rear customization microfluidic device, allows users to provide the setting of multiple panel able to programme.This of the present invention A little and other embodiments and its many advantages and feature contact and have hereafter carried out in more detail with accompanying drawing Description.

Accompanying drawing explanation

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 showing check valve shown in Fig. 2 A.

Fig. 3 is the letter of the method for a kind of operation microfluidic device according to an embodiment of the invention Change flow chart.

Fig. 4 is the method for a kind of operation microfluidic device according to another embodiment of the invention Simple flow figure.

Fig. 5 is a kind of operation method according to the microfluidic device of further embodiment of the present invention Simple flow figure.

Fig. 6 A is the sketch of reative cell in array according to an embodiment of the invention.

Fig. 6 B is control line in array according to an embodiment of the invention, fluid input line and anti- Answer the simplified perspective view of room.

Fig. 7 shows a kind of to programmable microfluidic device programming according to an embodiment of the invention Method for simplifying.

Fig. 8 shows a kind of to programmable microfluidic device volume according to another embodiment of the invention The method for simplifying of journey.

Fig. 9 shows a kind of to the programmable microfluidic device volume according to further embodiment of the present invention The method for simplifying of journey.

Figure 10 is the simplification signal of programmable microfluidic device according to an embodiment of the invention Figure.

Figure 11 is the rough schematic view of microfluid system according to an embodiment of the invention.

Detailed description of the invention

Fig. 1 is the rough schematic view of the microfluidic device according to one embodiment of the invention.At one In specific embodiment, the microfluidic device shown in Fig. 1 comprises Programmable high-density digital array. Described microfluidic device 100 comprises the first pressure source 102 and the second pressure source 104.In FIG, First pressure source 102 is referred to as accommodating accumulator (containment accumulator), as hereafter Described in more detail, described pressure source 102 is in fluid communication with control line, described control line operable in order to Close the valve being associated with reative cell (not shown) in array 106.Owing to using pressure source 102 Starting of carrying out makes these reative cell valves close and make sample and/or reagent to be contained in described reative cell In, pressure source 102 is referred to as accommodating accumulator the most in some embodiments.

In operation, liquid is placed on acute build up of pressure device (pressure accumulator) or pressure In power source, described acute build up of pressure device or pressure source subsequently with outside malleation supply (as containing compressed air Container) be connected.Under stress, described compressed air or other fluids are by described acute build up of pressure device Liquid push control line, thus start valve.Therefore, described pressure source does not generally comprise during fabrication Pressure fluid, and during being to provide for operating microfluidic device, pressure fluid can be accumulated wherein and store Upon activation control line to be executed stressed container.As being described more fully in text of the statement, Described acute build up of pressure device is operable to maintain the pressure in control line upon activation.At CO-PENDING with common It is suitable for carrying out embodiment party of the present invention with assigned U.S.Provisional Patent application No.61/044,417 provides The additional description of the digital array of case, all purposes are incorporated by by the entire disclosure by entirety Herein.

Fig. 6 A is the sketch of reative cell in array according to an embodiment of the invention.As one Example, some embodiments of the present invention use the cell with reative cell, and its lateral dimension is 100 μ m 60 μm, height is 125 μm.In this exemplary embodiment, the volume of described room is about For 0.75nl.The least building volume can use less sample volume and decrease running cost.Directly Through hole (via) the coupled reaction room of footpath 50 μm and mensuration/sample input line.With 110 μm first Lateral room spacing and the second spacing placing response room, lateral room of 200 μm.With containing mensuration/sample Thering is provided the valve of 50 μ m 50 μm in the layer that the layer of input line is different, described valve is operable in order to stop stream Through described input line.The particular geometric profile of this cell is not intended to limit embodiment of the present invention, And only it is to provide the example of specific embodiments.In another embodiment, employing is adapt to Other device geometric shapes of application.

According to some embodiments of the present invention, use width range be about 5 μm to about 400 μm, Depth bounds about 5 μm to the input line of about 75 μm provides and flows through the fluid of described microfluidic device. Width range is about 5 μm to about 400 μm, depth bounds is about the control line of 5 μm to about 75 μm Flow through the fluid of input line for closing (valve off) by valve.In some embodiments Width range is about 10 μm to about 500 μm, length is about 10 μm to about 500 μm, highly in employing Scope is about the reative cell of 5 μm to about 500 μm.The geometric shape of these devices side by example Formula provides, and is not intended to limit the embodiment described herein.

As shown in Figure 6A, it is provided that multiple input lines 620, fluid is made can to flow through institute in the horizontal direction State input line.Show three input lines, but some embodiments of the present invention use more than three Input line, such as, 11 input lines.Use a plurality of input line then single sample can be distributed in multiple In input line, this provides multiple copies of given sample.Such as more complete the retouching that Fig. 6 B is carried out Described in stating, described input line or input channel are at least partially embodied in the ground floor of microfluidic device In.Seeing Fig. 1, the mensuration input line 132 on the left of described input line and array 106 is in fluid communication, And be in fluid communication with the sample input line 142 on the right side of array 106.Therefore, mensuration input is derived from 130 or sample input 140 fluid be all provided to input line and and then be supplied to reative cell.

There is provided multiple control line 610 in vertical direction, enabling control the fluid through input line Flowing.Show two input lines, but some embodiments of the present invention use the control more than two Line, such as, 70 control lines.In another embodiment, there are 71 control lines.Described many Bar control line forms, along the length of input line, the reative cell separated, it is provided that comprise the multiple of same sample Reative cell.As described in the description more complete to Fig. 6 B, described control line or control passage are at least It is partially contained in the second layer of microfluidic device.See Fig. 1, described control line and lock-in control line 114 partial fluid communication of 110, described lock-in control line is in fluid communication with accommodating accumulator 102. Intersection point at described control line with described input line forms valve 615, and it responds to the fluid in control line Pressure and actuated, and operable with stop fluid flow through described input line.Generally, described multilamellar is micro- Fluid means comprises multiple elastomer layer and valve 615 comprises deflectable diaphragm.Show at Fig. 6 A and 6B In the embodiment shown, the deflectable diaphragm of described valve deflects to be positioned at and controls on the intersection point of passage In fluid passage.Therefore, shown embodiment uses " above pushing away " valve, wherein deflectable diaphragm to Upper deflecting in fluid passage with in closedown fluid passage, valve position.For at Fig. 6 A and 6B The valve of middle display, fluid pressure causes deflectable diaphragm to return to non deflected position present in release control passage Put thus the valve of either on or off.

Flow through the fluid of input line 620 along the direction vertical with Fig. 6 A plane through through hole 625 and to Upper inflow reative cell 630, such as reative cell 630 as described in being described more fully Fig. 6 B It is at least partially embodied in the third layer of microfluidic device.Therefore, described through hole is at least partially embodied in The most described the second of microfluidic device or third layer in.Generally, laser ablation is used to process removal the Two or the part of third layer form described through hole.Owing to described microfluidic device is that air can pass through , closing filling technique (blind fill technique) filling reative cell can be used and carry out multipleization , biological or other experiments.Skilled in the art realises that, after there is fluid in the reaction chamber, control Starting of line processed will cause valve close and fluid retains predetermined time period in the reaction chamber.

Fig. 6 B is the control line in the array of one embodiment of the invention, fluid input line and reaction The simplified perspective view of room.Array (such as, the array 106 shown in Fig. 1) is multilamellar microfluidic device A part.Every layer generally comprises the elastomer knot with one or more groove, passage, room etc. Structure.As shown here, ground floor 601 comprises the multiple control passages being arranged to array of parallel channels 610 and other control passage 611.Described control passage 610 and other control passage 611 with One or more pressure source or acute build up of pressure device fluid communication.The most in one embodiment, control Passage 610 processed is in fluid communication with acute build up of pressure device 102, control passage 611 with acute build up of pressure device 104 fluid communication.Although show single control passage 611 in fig. 6b, but the common skill in this area Art personnel are it should be understood that shown single passage represents one or more that be associated with 601 layers Control passage.

Additionally, described control passage is not restricted to the position relevant to layer 601.Described control passage can It is adapted to specifically apply and be placed in other layers.Such as, at a multiple control (control-on-control) in enforcement, in order to make the first control line (such as controlling passage 611) Second control line (such as controlling passage 610) is produced and controls, by described second control line according to control The length of passage processed is placed in both ground floor 601 and the second layer 602.Use and through hole 625 phase As through hole the second control line is transformed into the second layer 602 from ground floor 601.By controlling first Line passes over the second control line, and the intersection point at two control lines forms valve.Once start described first Control line, between the most described first control line and described second control line, the flexible membrane of valve position is pushed up Dynamic to block the second control line being positioned in 602 layers at valve position.Therefore, as the most complete Describe, " above pushing away " valve can be formed between different control lines or between control line and fluid input line. Seeing Fig. 1, the control line being connected with valve 128 is transitioned into upper strata by through hole from the lower floor of structure, and Control line 122 is made it is thus possible to start separator valve 124 afterwards through control line 122.Then, with valve 128 The control line being associated through another through hole wear return in ground floor thus pass through and start fluid input The valve of line 132.Once control line 122 actuated, and separator valve 124 blocks through being connected with valve 128 The flowing of control line, stop valve 128 generally to respond to the closedown that control line 110 starts.

The second layer 602 comprises the multiple fluid inlet channel 620 also arranged with array of parallel channels.? In embodiment shown by Fig. 6 A and 6B, control passage 610 and input channel 620 is hung down each other Directly arrange.By liquid (the most of a substantially incompressible stream present in control passage 610 Body) apply pressure and realize starting of valve present in microfluidic device.Generally, liquid is placed on The fluid of pressurization is provided (such as in accumulator (accumulator) or pressure source and to accumulator Air, nitrogen etc.).In accumulator, the increase of pressure makes liquid enter control line under stress.Response In pressure applied, the deflectable diaphragm forming 601 layers of top upward deflects entrance fluid inlet channel 620.Therefore, passage and fluid inlet channel intersection point generation " above pushing away " valve are being controlled.Some other Embodiment can by reorientating various control and fluid layer and use " under push away ", " drop-down " or " on Draw " valve.

In some embodiments of the present invention of multiple control are provided, it is possible to provide one or more is another Outer key-course (such as lower floor 601), or by control line being arranged into 602 layers.Described one Individual or more other key-courses can comprise other control line (not shown), its once start by By control line present in valve closed layer 601.Therefore, by the control other to one or more Controlling fluid (such as liquid) present in line processed and apply fluid pressure, flexible membrane deflects to control line 610, stop fluid to flow through control line.Therefore, some embodiments of the present invention provide key-course, Also referred to as multiple control.Those of ordinary skill in the art are it will be appreciated that many changes, improves and replace.

The described second layer also comprises multiple through hole 625, produces and provides from described input channel to third layer The fluid passage of the fluid flowing of 603.Third layer 603 comprises multiple reative cell 630, and it is by logical Hole is in fluid communication with described fluid inlet channel.In the embodiment illustrated, reative cell 630 is formed For being connected with the bottom of layer 603, in other words, described reative cell is open from bottom.Therefore, logical Hole is completely contained in 602 layers.In other embodiments, described through hole can be comprised in To be adapted to concrete application in both 602 and 603 layers.

In some embodiments, microfluidic device can comprise according to rotate or cast fabrication scheme prepare One or more layer.Such as, rotation approach can relate to place on figuratum dish or mould gather Condensation material, and rotate described dish to produce polymeric layer on dish.Illustrative polymers includes poly-methyl Acrylate, polystyrene, polypropylene, polyester, fluorinated polymer, politef, poly-carbonic acid Ester, polysilicon and polydimethylsiloxane (polydimethylsiloxane, PDMS).Cast side Case can relate to as being cast in by PDMS material on figuratum template or mould, and this can produce can be from The PMDS layer intactly peeled on mould or leave behind.Generally, prepared by cast manufacturing technology Thickness prepared by layer ratio rotation manufacturing technology.Elastomer block can comprise one of any required combination or more Multiple cast or rotating layer.

In some embodiments, ground floor 603 can be manufactured according to rotation approach.Such as, can be by PDMS is placed on to be had on the mould corresponding to the multiple required bossing controlling passage 610. Rotatable described mould is to produce PDMS thin layer on whole mould.After hardening, can be from mould On peel ground floor 601 off and be attached to applicable rigid matrix (such as glass, silicon) or plastics (polyphenyl Ethylene) on.Or, ground floor 601 can remain and be attached on mould.Ground floor 601 can comprise Opening, groove or be at least a partially formed or limit control passage 610 other spaces.

In order to generate the second layer 602, rotation approach can be used, PDMS is placed on the second mould, Described second mould has the bossing corresponding to multiple required fluid inlet channel.Second mould is also Can comprise, such as, form projection or the wavelike segments corresponding to arranging labelling in the second layer 602.These Arrangement labelling can use in the laser ablation for forming through hole 625, so that swashing during ablation processes Described arrangement labelling is pointed in light ablation.Rotatable second mould is to provide on whole second mould PDMS thin layer.The second layer 602 can comprise opening, groove or to be at least a partially formed or limit input logical Other spaces in road 620.In some cases, the second layer 602 can be exposed on one or many State laser ablation.The ablating laser beam pointing to the second layer 602 can form through hole 625.At the second layer 602 Fully after solidification, ground floor 601 can be peeled off from the second mould, and it is alignd with ground floor and connects Touch.The second layer can be pasted so that two-layer sticks on rigid matrix with ground floor.

In order to use casting program to produce third layer 603, PDMS can be cast in and have corresponding to many On 3rd mould of the bossing of the reative cell 630 needed for Zhong.After hardening, can be from the 3rd mould On peel third layer 603 off and align with the second layer 602 and contact.Third layer can be pasted onto the second layer It is upper so that all three layers stick on rigid matrix.The material that can manufacture microfluidic device includes but does not limits In elastomer, silicon, glass, metal, polymer, pottery, inorganic material and/or these materials Combination.

Referring back to Fig. 1, pressure source 102 is in fluid communication with closing control line 110.Described lock-in control Line comprises several section being hereafter described more fully and check valve 112 (also referred to as check-valves (check valve)).At CO-PENDING and commonly assigned international patent application No.PCT/US07/080489 (open with international publication number WO 2008/043046 A2, the entire disclosure is drawn by entirety With being expressly incorporated herein) in provide the other description about check valve.First section 114 is from pressure source 102 receive and start pressure and operable accommodate valve present in closed array 106 in order to closing (containment valve).As understood by the skilled person, array 106 is adapted for Multiple micro-fluid experiment.Thus receiving can be closed by responding to starting of lock-in control line 110 Valve so that present in the reative cell in array, sample, reagent etc. retain in the reaction chamber.Locking control The section 114 of line 110 processed is without check-valves, and this makes receiving valve start pressure remove pressure source 102 After reopen.

Second section 116 of lock-in control line 110 is in the downstream of check valve 112.Due to check valve 112 Operable in order to stop fluid to flow to section 114 from section 116, starting of pressure source 102 will cause Control fluid (normally liquid) and by check valve 112 and close valve closing 118a-118e.Institute in Fig. 1 The fine rule being associated with lock-in control line 110 shown represents " overhead (flyover) " district of control line Section, wherein the design of control line prevents mensuration input line 132 (being described more fully hereinafter) from existing Lock-in control line 110 is closed after starting.The thick line being associated with valve 118a-118e represents and can be operated In order to close or to disable manifold 135 (providing the fluid flowing between multiple mensuration input lines 132) Valve.Close to stop and measure stream through manifold 135 from a mensuration input line in the pass of valve 118a-118e Flow to other mensuration input line, this will be referred to as disabling of manifold.

After starting lock-in control line 110 by pressure source 102, valve 118a-118e can close and Remove lock-in control line start pressure after, check valve 112 can keep the closedown of valve 118a-118e State.Contrary with valve 118a-118e, section 114 the receiving valve started can start described in removing Open after pressure.Therefore, in lock-in control line, the locus of check-valves provides with locking or non-closes One or more section that lock behavior is characterized.As those skilled in the art are clear and definite, at control line The extra check-valves of middle employing can provide the extra section with locking behavior.

Lock-in control line 110 also comprises the 3rd section 128, its have be operable to close from survey Surely the valve of the mensuration input line 132 of input 130 to array 106.Measure input 130 may be additionally referred to as Input port.Due to the 3rd section 128 also in the downstream of check valve 112, started by pressure source 102 Lock-in control line 110 can be closed the valve in the 3rd section 128 and stop in mensuration input line 132 and battle array Row 106 and on it arrangement reative cell between flowing.Removing starting of lock-in control line 110 After pressure, the valve of section 128 remains turned-off.

Thering is provided and the second pressure source 104 of the second control line 120 fluid communication, it is referred to as interface and stores Long-pending device (interface accumulator).Described second control line also comprises several section 121,123 With 125.Section 121 comprises elevated region 122 and enables to do not closing mensuration input line 132 In the case of start separate valves 124.Provide in this specification to valve 124 (referred to herein as Independence or separator valve (independent or decoupling valve)) other description, and under It is described more specifically by literary composition.Although specific independence described in FIG or separator valve 124 Use the second pressure source 104 to start, but this is the most optional.Implement at other In scheme, independence or separator valve 124 can be mechanical, electrostatic, fluid, electromechanical, heat Power, piezoelectricity etc..Therefore, start point although with the second pressure source 104 of display in Fig. 1 Every valve 124, but this is the most optional.Although additionally, using single Two pressure source 104 controls the multiple valve groups comprising separator valve, and other embodiments can use many Combination and electrostatic that individual pressure source fluid starts start.Those of ordinary skill in the art understand multiple change Change, improve and replace.

Section 123 comprises and is operable to close in 6 shown mensuration input lines 132 5 Valve 134.Owing to only providing 5 valves 134 in the embodiment depicted in fig. 1, pass through pressure source In the section 123 that 104 are carried out, starting of valve 134 still can make the mensuration input line of low order end be held open. Section 125 comprises check valve 127 and is operable to close the valve 126 of sample input line 142, Described sample input line 142 is in fluid communication with sample input (i.e. input port) 140 and array 106. It is noted that while show that mensuration and sample input in FIG, the invention is not restricted to only measure and sample Product input, and other inputs are included in the scope of the present invention.

Unidirectional or check-valves 127 are placed on the section 125 of lock-in control line 120, it is allowed to 126 valves Close and remain turned-off after section pressure applied, stoping fluid by sample input line dealing array The flowing of 106.Owing to section 121 and 123 does not comprise check-valves, the cancelling of these sections start by Cause reopening of valve 134 and 124.

Some embodiments of the present invention provide asynchronous logic function in microfluidic device 100.Example As, owing to using the control line 120 individually and independent of control line 110 shown in other to start in Fig. 1 Shown independence or separator valve 124, some embodiments of the present invention provide multiple control.At figure In 1, lock-in control line 110 applied before starting separator valve 124 pressure locking can accommodate valve 128.On the other hand, before lock-in control line 110 being applied pressure by receiving accumulator 102, Start lock-in control line 120 by interface accumulator 104 thus close separator valve 124 and prevention is accommodated The closedown of valve 128.In other words, if separator valve 124 cuts out when starting to apply pressure to valve 128, Then the closed mode of separator valve stops pressure applied to arrive valve 128.Therefore, control line 110 He The order of starting of 120 causes different valve to be operated, and this provides asynchronous logic.

The multiple control that embodiment of the present invention provides makes array 106 to be " programmed ".Such as, Owing to array 106 is by the closedown of multiple valves, locking or the sequential programming reopened, can be with multiple cloth Office uses array 106.In embodiments more as herein described, use shown 2 pressure source Provide three kinds of different layouts.(it can be considered stacking control valve group to use check valve and separator valve In one) these multiple layouts are provided.Separator valve can limit the control to valve 128, described control Can be by responding to the realization of starting of lock-in control line 110, described separator valve simply stacks control valve One example, the scope of the invention includes other layouts.

Predetermined segment at control line uses check-valves to provide the part locking of control line.Such as, may be used Locking-valve 118 and 128 is started by initial control line 110.But, initial control line 120 Start the closedown that can cause separator valve.Subsequently control line 110 start the locking that can cause valve 118 But on valve 128 without impact.Cancel while keeping control line 110 to start subsequently and start control line After 120 (pressure such as applied by release interface accumulator) can cause separator valve to reopen The locking of valve 128.

In the microfluidic device 100 shown in Fig. 1,6 measure input line 132 and are each separated into 4 Input line, it is provided that through 24 the mensuration input lines altogether accommodating valve 128.Additionally, show 24 sample input lines 142.The concrete numeral simply example of these input lines, other embodiments carry Supply the input line number that other are concrete.Such as, in a specific embodiment, showing shown in Fig. 1 It is intended to illustrate only half microfluidic device (right side of such as device), has in the left side of described device The input group matched.Those of ordinary skill in the art are it will be appreciated that many changes, improves and replace.

Fig. 2 is the simplification sectional view of the check valve of one embodiment of the invention.Fig. 2 B is Fig. 2 A The simplification top view of shown check valve.See Fig. 2 A, start fluid (normally liquid) through entrance 210 flow into check-valves 112/127, and flow through room 220 through through hole 215.By starting the stream of fluid Move and film 230 is mentioned or is maintained at the position of basic horizontal, make to start fluid through entrance 210 From left to right pass.Start fluid via through holes 240a-240d and through hole 242 flows through room 220 and from going out Mouth 250 flows out.Therefore, in the embodiment illustrated, for each check valve 6 through holes altogether For, there are an input through hole and 5 output through holes.As shown in the top view at Fig. 2 B, root Extra structural detail and assembly is provided according to specific design.

When fluid pressure is started in removal, film 230 is collapsed upon on layer 260, stops by entrance 210 Backflow.Therefore, check valve 124/127 provides from entrance 210 to the flowing of outlet 250, but stops Reverse flowing.It is described in CO-PENDING about other of check valve and commonly assigned above-mentioned quotes Application provides.

Some embodiments of the present invention provide integration the unidirectional or microfluidic device of check-valves.? In one embodiment, described microfluidic device comprises the first valve and the second valve.Generally, described first Valve is in the multiple valves forming the first valve group, and described second valve is the multiple of formation the second valve group In valve one.Fill as microfluid with the control line of described first valve and described second valve fluid communication The part put provides together with pressure source, and described pressure source is in fluid communication with described control line.See Fig. 1, starts control line 110 by receiving accumulator 102 is applied pressure, causes in array 106 Valve 615 and valve 118a-118e starts so that manifold 135 disables.Described microfluidic device is also Comprise the check valve in the control line being arranged between described pressure source and described second valve.Such as, rise The valve 112 just opened is on the control line 110 accommodated between accumulator 102 and valve 118a-118e. After reducing or removing the pressure on control line 110, check-valves 112 provides closing of valve 118a-118e Lock, and present in array 106 valve (non-blocked valve) can reopen make sample flow into reative cell. Therefore, by using the valve of predetermined number in check-valves locking microfluidic device, it is provided that be commonly designed In the control of fluid flowing and the separation of sample in irrealizable microfluidic device.

In the embodiment shown in Fig. 1 and 6A, valve 615 is arranged to spaced apart at microfluid Reative cell on device, and valve 118a-118e is arranged to be connected with mensuration input port 130 (coupled) six lines that the first fluid input line of six lines is connected with mensuration input port Second fluid input line separate mutually.As being described more fully in this manual, close and locking-valve 118a-118e so that the ability that manifold 135 disables enables microfluidic device to program in many ways, This makes the fluid flowing in input line 132 can be separated according to concrete application or merge.

In addition to being integrated into by check-valves and having in the microfluidic device of array layout, some designs carry Supply separate valves, also referred to as separator valve.Such as, the controlled separation independent of other valves in device It is logical that valve is operable to stop starting of one or more valve, one or more valve to control Cross with microfluidic device present in the fluid flowing of the fluid input line that is connected of reative cell.Such as, may be used Before starting control line 110, close separator valve 124, thus stop valve 128 to respond to control line 110 The closedown started.

Figure 10 is the rough schematic view of the programmable microfluidic device of one embodiment of the invention.Figure Element shown in 10 can provide in the carrier or fill at the microfluid being attached at carrier as shown in figure 11 Middle offer is provided.As shown in Figure 10, the profile of the microfluidic device being loaded on carrier is by dotted line 1050 Represent.Seeing Figure 10, the opposite side at microfluidic device provides two emission sources (vent source). The emission source provided usually used as a part for carrier can not use in all embodiments.At figure 48 sample input lines 1010 shown above of 10, wherein the left side at microfluidic device arranges Article 24, sample input line and on the left of array 106 provide sample, arrange on the right side of microfluidic device Article 24, sample input line and on the right side of array 106 provide sample.Article 48, sample wire is the most in the carrier There is provided and by the through hole 1030 formed in microfluidic devices and align with sample wire end and battle array Row 106 are in fluid communication.There is provided the sample from through hole 1030 to array 106 defeated in microfluidic devices Enter line 142.Article 48, sample wire (is not shown by UP source (common pressure source) Show) pressurization, the sample part of the carrier that described pressure source can be placed on it with microfluidic device is connected. Therefore, the sample of 48 sample ports can be written into microfluidic device and push array through sample input line 106.See Fig. 1, for the sake of clarity show the right part of array 106, wherein Figure 10 24 sample input lines of right part are shown as sample input line 142.Therefore, the one of the present invention A little embodiments provide 48 sample input lines with up to 48 different samples.Other are implemented Different numbers is provided according to concrete application.

Showing control line 1004 in Fig. 10, the interface accumulator 104 that it shows with Fig. 1 is connected. Additionally, show several check-valves (check valve, CV) in Fig. 10, this is at microfluidic device In provide the one-way fluid flow through control line.See the bottom of array 106, for being connected to array The control line of this part provides other CV.Aquation line is provided in the outside of array 106.This A little aquation lines are that aquation fluid provides source, and the effect of aquation fluid is to reduce or prevent to flow in array The evaporation of body.

48 input lines shown in Figure 10 bottom comprise 12 and measure input line (array 106 left side 6, side, on the right side of array 106 6).Article 12, measure line to be added by UP source (not shown) Pressure, the mensuration part of the carrier that described pressure source can be placed on it with microfluidic device is connected.Therefore, 12 measure mouths mensuration can be written into microfluidic device and after measured input line push 2 arrays 106. See Fig. 1, for the sake of clarity show right array 106, its have be shown as measure input line 132 6 sample input lines of Figure 10 right part.Therefore, some embodiments of the present invention provide There are up to 12 different 12 the mensuration input lines measured.Measure input line at microfluidic device Two arrays 106 of central authorities between pass through, 6 lines are branched off into 106,6 lines of right array and are branched off into Left array 106.The use of shown manifold is provided that less than 12 mensuration, such as, few to single survey Fixed.Other implement to provide different numbers according to concrete application.

In 36 input lines of other shown in Figure 10 bottom, 2 lines are for aquation, concrete at this In enforcement, 34 is untapped.Measure input line 1020 and the single input entering array 106 is provided The input of line, and measure input line 1022 and be branched off into the input line of a plurality of entrance mensuration 106 (such as, Article 4, line).

Fig. 6 A shows the example of every 3 input lines of sample, but in the embodiment shown in Figure 10 In, there are 11 input lines.Additionally, in fig. 6,2 control lines show in fig. 6, but In the embodiment depicted in fig. 10, have 71 control lines can be operable to form 70 reative cells/ Input line.Therefore, there are 770 reative cells (11 defeated for each sample in the present embodiment Enter line × 70 reative cell).Along with 48 samples are dispensed into 770 reative cells, it is provided that highly dense The integrated fluid loop (integrated fluidic circuit, IFC) of degree.In one embodiment, Up to 48 independent samples of microfluidic device energy one-time detection.Described up to 48 samples each It is dispensed into 770 reative cell independence groups, thus delivers the synchronous digital PCR if sum is 36,960 Reaction.In another embodiment, some programmability of microfluidic device are removed, then for phase Same device area and size of components, add the reaction number of chambers/sample.Such as, an embodiment Using up to 48 samples, it has 814 reative cell/samples.

Use the product being available from assignee of the present invention, can complete whole in less than 4 hours Digital pcr process.Additionally, microfluidic device as herein described is complete gene alaysis system Ingredient.This system can comprise microfluidic device, the controller of microfluidic device, BioMark^TM System or integral type thermal cycler, EP1 reading instrument and related software.Microfluidic device as herein described Compatible with the distribution design of finished product reagent and cellular form.Those of ordinary skill in the art understand many and change Become, improve and replace.

Figure 11 is the rough schematic view of the microfluidic device of one embodiment of the invention.Described miniflow System system comprises carrier 1100 and is loaded on the microfluidic device 1108 on described carrier.Microfluid fills Put 1108 and incorporate the element described in Fig. 1.Described carrier comprises and is arranged in platform (bank) 1106a On multiple first input ports or hole 1105.Described carrier also has and multiple first input port fluids Multiple first input lines 1115 (such as, 48 input lines) of connection.Input the plurality of first There is outer peripheral edge around mouthful, allow to UP source described first input port pressurization.Show at one In example application, 48 samples are loaded into the first input port, from the top of carrier, described mouth are applied Pressure causes 48 samples to be pushed over multiple first input line the stream eventually entering on microfluidic device Body line.

Described carrier also comprises multiple second input ports 1105 being arranged on platform 1106b and multiple The second corresponding input line 1115.It is similar to multiple first input port, the plurality of second input port By outer peripheral edge around, enable the fluid being dispensed into the second input port to be pushed over the second input line and to enter Fluid line in microfluidic device.

In order to provide fluid communication between described carrier and the microfluidic device being placed on described carrier, Microfluidic device has the through hole 1114 formed in microfluidic device bottom, itself and described first input The end section alignment of line and described second input line.The fluid flowing through input line passes through hole to enterprising Enter the fluid line that microfluidic device provides.As shown in Figure 10, the fluid of described sample input port is connected Line enters microfluid dress through the through hole 1030 arranged with Right vertical on the left of microfluidic device along figure The offside put, and flow to the central authorities of microfluidic device.In fig. 6, these sample input lines can be right Answer the input line 620 that in Fig. 6 A, level flows through.Use this 48 sample input lines, can be at microfluid The reative cell that device is often gone is filled different samples.

The bottom of microfluidic device additionally provides through hole 1040 and it is with multiple second input lines 1020 alignment.As shown in Figure 10, being loaded into the fluid (as measure) of the second input port, to flow through second defeated Enter line 1020, enter manifold 1060 through through hole 1040.Described microfluidic device comprises can be with described The crew-served valve of manifold, to cause 1 fluid (sample or mensuration) or 12 different fluids It is provided the reative cell in two arrays.If application needs single input fluid, manifold is unlocked And input fluid flow in Figure 10 through array centre perpendicular through all fluid lines.Fluid line Branching out through array centre, finally flowing through the through hole 1030 of array side along with them Flow through reative cell eventually.Seeing Fig. 6 A, these measure input lines can corresponding process defeated under reative cell Enter line 620.In this example, the flowing of left array side is from central authorities to left side, the flowing of right array It is to right side from central authorities.

Or, manifold 135 can be closed to introduce the different fluid of 12 reative cells to be introduced (such as Measure) (6 different mensuration of the most each array).As shown in Figure 10, fluid hangs down between array Direct current moves, and then branch goes out, wherein each inflow array of 12 fluids multiple rows of in, level Ground passes through to the neighbouring through hole in edge 1030 from central authorities.

Carrier provide acute build up of pressure device 1106c and 1106d enable to start microfluidic device Present on control line and check-valves.Control line 1002 in the way of similar with fluid line, in carrier With 1004 by the through hole formed on microfluidic device and the control line 110 and 120 on microfluidic device Fluid communication.Therefore, while figure 1 show that the interface of carrier and receiving accumulator and microfluid fill Control line in putting, it should be understood that this schematic diagram is the sketch for the sake of clear and convenient.With can be thereon Other details that the carrier of placement microfluidic device is correlated with are in U.S. Patent Application Publication Thering is provided in No.2005/0214173, the disclosure of which is incorporated by this for all purposes by entirety Literary composition.

Carrier 1100 has the acute build up of pressure device hole 1101 and 1102 of integration, each of which has for Accommodating the dry hole 1103,1104 of valve, described valve is preferably the check-valves being connected with lid.Carrier 1100 Also comprise one or more hole platform 1106a, b, c and d, each of which have be located therein or Multiple holes 1105 (also referred to as input port).The hole 1105 of each carrier 1100 has from hole 1105 Guide the passage of the microfluidic device 1108 being placed in carrier positions 1107.Hole platform 1106c and 1106d It is typically used to provide the pressure of control line in the presence of starting microfluidic device 1108.There is provided Control the fluid line of fluid, valve present in the hole of Kong Taizhong and microfluidic device or other control are set For being connected.Microfluidic device is preferably the elastomer formed by two-layer or more layers elastomeric material Block, its groove with the micro-manufacture formed wherein or passage.

Described microfluidic device have one or more with what one or more through holes 1114 were in fluid communication Passage, it provides the fluid communication between the passage in microfluidic device and the passage in carrier then, Then the hole 1105 in its pilot hole row 1106a-d, thus provide hole in carrier 1,100 1105 with The fluid communication between passage in microfluidic device 1108.Accumulator bore top 1109 and 1110 is even Receive accumulator bore 1101 and 1102 to form accumulator room 1115 and 1116.Accumulator bore top 1109 Comprise valve 1112 and 1111 respectively with 1110, it is preferably and introduces a gas into accumulator under stress Room 1115 and 1116 and retain the check-valves of gas.When being present in accumulator room 1115 and 1116 Time, valve 1111 is positioned at the inside of dry hole 1102 and 1104 with 1112 so that liquid does not contact valve 111 With 1112.Preferably by mechanically opening by laminated flake (shave), pin etc. in preferred check-valves Open valve 1111 and 1112, its overcome the power that is automatically switched off of check-valves to discharge pressure from accumulator room, Thus reduce fluid pressure contained in accumulator room.

Carrier 1100 and its associated component can be by polymers manufacturings, such as polypropylene, polyethylene, poly-carbon Acid esters, high density polyethylene (HDPE), polytetrafluoroethylene PTFE or Teflon (R), glass, quartz, gold Belong to (such as aluminum), transparent material, polysilicon etc..Accumulator bore top 1109 and 1110 also comprises tune Whole screw, it is removable to introduce from accumulator room 1115 and 1116 or to remove gas or liquid.Excellent Selection of land, can start valve 1112 and 1111 with release accumulator room 1115 and 1116 in fluid pressure, Otherwise fluid pressure then retains wherein.Notch 1117 is used to help to place microfluidic device suitably In other instruments, such as, for operating or analyze microfluidic device or the instrument of reaction wherein carried out Device.

Fig. 3 is the letter of the method showing operation microfluidic device according to an embodiment of the invention Change flow chart.In the embodiment depicted in fig. 3, it is provided that 48 panel layouts of digital array. Pressure source 102 (receiving accumulator) is applied the first pressure, which results in lock-in control line 110 (310) Start.In a specific embodiment, described first pressure is 30psi.Real at other Execute in scheme, use other pressure according to concrete application.See Fig. 1, all valves in array 106 (not shown) can be closed due to the pressure of section 114.Pressure is applied to district by check valve 112 Section 116, this closedown that can cause valve 118a-118e and locking.The overhead portion of section 116 can hinder Only measure input line to close due to starting of section 116.Close and close input fluid in the pass of valve 118 Through the flowing of manifold 135, thus 6 the mensuration input lines being connected to measure input 130 are divided each other Every.

It should be noted that in the control stream shown in Fig. 3, do not use pressure source 104.Therefore, do not control Starting or the closedown of separator valve 124 of line 120.Therefore, respond to starting of lock-in control line 110, Accommodate valve 128 to close and locking.Therefore, the first pressure is being applied (such as, by receiving accumulator After 30psi), described array valve cuts out and valve 118 and 128 cuts out and locking.

The first pressure (312) reducing pressure source 102 makes array valve (not shown) reopen. In some embodiments, in process (312), pressure is removed so that pressure applied is zero. As noted above, valve 118 and 128 can keep locking in off position.Be supplied from sample input or The sample of mouth 140 is loaded into the reative cell in (314) array by sample input line 142.Chip Design provides by sample input line 142, measures input line 132 or both loadings.Because at this In embodiment, measure input line and closed by locking-valve 128, load and carry out through sample input line. After completing sample loading, pressure source 102 is applied the second pressure (316) again, thus closes closed array Valve also separates the sample in described reative cell.In some embodiments, the second pressure is equal to the first pressure Power (such as, 30psi), although this is the most optional.Other are suitable to close closed array valve Pressure within the scope of the present invention.Although additionally, removing first in some embodiments Pressure, this is the most optional, because some embodiments can be by pressure drop as little as non-zero Value and still provide load sample sufficient fluid flowing.

Therefore, the embodiment that Fig. 3 shows provides the loading of 48 samples (that is, shown in 24 Sample input line × device both sides).As discussed below, programmable digital battle array as herein described Row also provide other sample layout.

It should be understood that the concrete steps shown in Fig. 3 provide the miniflow of operation one embodiment of the invention A kind of ad hoc approach of body device.Other sequence of steps are real also dependent on some alternate embodiments Execute.Such as, some alternate embodiments of the present invention can be implemented listed above in a different order Step.Additionally, single step shown in Fig. 3 can comprise many sub-steps, it can be according to single step Requirement implement with multiple order.Additionally, can add according to concrete application or remove other step. Those of ordinary skill in the art are it will be appreciated that many changes, improves and replace.

Fig. 4 is the side of a kind of operation of display microfluidic device according to another embodiment of the invention The simple flow figure of method.Different from 48 panel layouts described by Fig. 3, in the enforcement shown in Fig. 4 Scheme provides single sided board layout.Pressure source 104 applies the first pressure (410), and this causes closing Starting of lock control line 120.Pressure can apply to close and locking section 125 by check valve 127 In valve 126.Therefore, the flowing through sample input line 142 is not the most had.Overhead Section 122 can transmit control pressure to close separator valve 124.

Control pressure transmission in lock-in control line 120 can be measured input line to 6 by section 123 5 valves being associated 134 in 132.Therefore, valve 134 will be closed so that flowing only passes through The mensuration input line of low order end.

Mensuration input line loading single sample (412) through low order end.Due to lock-in control line 110 Section 116 does not actuated in this layout, and manifold 135 is opened, and makes in the input of low order end sample contained Single sample be provided to whole sample input lines 132.Therefore, use from low order end sample The single sample measuring input inputs the described array of loading after measured.Acute build up of pressure device 102 is applied Two pressure, start lock-in control line 110 and close the array valve being connected with section 114.Therefore, make With different control stream, the multiple layout using single programmable digital array is possible, such as, Single sided board layout.

It should be understood that the concrete steps shown in Fig. 4 provide according to another embodiment of the invention micro- A kind of particular methods of operation of fluid means.Other sequence of steps are real also dependent on alternate embodiment Execute.Such as, some alternate embodiments of the present invention can be implemented listed above in a different order Step.Additionally, single step shown in Fig. 4 can comprise many sub-steps, it can be according to single step Needs implement with multiple order.Additionally, can add according to concrete application or remove other step. Those of ordinary skill in the art are it will be appreciated that many changes, improves and replace.

Fig. 5 is the side of microfluidic device showing a kind of operation according to further embodiment of the present invention The simple flow figure of method.In the embodiment depicted in fig. 5, it is provided that 12 panels of digital array Layout.Pressure source 104 is applied the first pressure (510).In a specific embodiment, institute Stating the first pressure is 45psi, although this is the most optional, can use other to be suitable for Pressure.Such as the discussion of the process (410) about Fig. 4, start lock-in control line 120.By list Pressure is applied with the valve 126 closed and in locking section 125 to valve 127.Therefore, in this layout Do not have the flowing through sample input line 142.Elevated bridge section 122 can transmit described control pressure with Close separator valve 124.As discussed below, the Temporarily Closed of separator valve can stop receiving valve 128 Close under the effect of starting of control line 110 subsequently.Section 123 can be by lock-in control line 120 Control pressure transmission and measure 5 valves being associated 134 in input line 132 to 6.Therefore, valve 134 can be temporarily closed.

Pressure source 102 is applied the second pressure (512), starts lock-in control line 110.At a tool In the embodiment of body, described second pressure is 30psi, less than the first pressure (such as, 45psi). As discussed below, described second pressure be enough to close predetermined valve and be not turned in process (510) The separator valve 124 of middle closedown.Other pressure can be used according to concrete application.Section 114 start meeting Close the valve in closed array 106.Pressure can apply to section 116 through check valve 112, and this can cause valve The closedown of 118a-118e and locking.The overhead portion of section 116 can stop mensuration input line due to district Section 116 start and close.Close and close the flowing inputting fluid through manifold 135 in the pass of valve 118, Thus by with measure input 130 is connected 6 measure that input line is each to be separated.

Start different from control line 102 initial, separator valve cuts out by initial the starting of control line 104, Thus provide programmable sequences, wherein starting of control line 102 is not resulted in and measures input line 132 Valve 128 locking being associated.This multiple control characteristic provides employing some embodiments of the invention Asynchronous logic function, use conventional microfluidic device will not obtain this characteristic.

It is applied to the pressure drop the most such as zero (514) of pressure source 102.Because section 116 is at list To valve 112 downstream, after section 116 is cancelled and started, valve 118 continues to be latched in closed position, this resistance The fluid flowing that only 6 measure between input line.Cancelling of section 114 is started in the described array of unlatching Valve, make in loading process subsequently load sample.As discussed above, valve 128 is in process (512) in non-latching.

It is applied to the pressure drop the most such as zero (516) of pressure source 104.By reducing section 123 Pressure reopen valve 134.As discussed above, mensuration input 130 is used to come in array Reative cell provide input.Again beaten by the pressure reduced in section 121 in elevated bridge section 122 Drive separator valve 124.Owing to control line 110 not applied pressure, institute in this stage of programming process It is maintained at opening with valve 128.Although shown for pressure drop is as low as zero, but these tool Body pressure is not essential to the invention, can use other pressure being suitable for.

Measure from 6 and input is loaded into sample (518).Caused by the locking due to valve 118 before Manifold 135 go functionalization keep measure input between separation.Therefore, in this layout, carry 12 panel layouts (6 2 sides measuring input × device) are supplied.Pressure source 102 is applied the 3rd The array valve that pressure (520) is associated with section 114 with closedown.Additionally, due to separator valve 124 It is switched on again, so valve 128 is blocked cut out.

It should be understood that the concrete steps shown in Fig. 5 provide operates the micro-of further embodiment of the present invention A kind of concrete grammar of fluid means.Other sequence of steps are implemented also dependent on alternate embodiment. Such as, the alternate embodiment of the present invention can implement step listed above in a different order. Additionally, single step shown in Fig. 5 can comprise many sub-steps, it can be according to single step with multiple suitable Sequence is implemented.Additionally, can add according to concrete application or remove other step.The common skill in this area Art personnel are it will be appreciated that many changes, improves and replace.

Table 1 provides the multiple of programmable digital array microfluidic device as herein described may panel One of layout start pressure and loading process.The pressure example that pressure source 102 and 104 is applied Property pressure illustrate, although other be suitable for press packet be contained in the scope of embodiment of the present invention.

Table 1

The integration of check valve and separator valve and digital array provides the single high-density digital array of use The multi-user of chip programs layout.Therefore, some embodiments of the present invention provide the saving of cost Raising with experiment flux.Serial described herein or asynchronous logic provide the apparatus function of enhancing, this It is owing to the state of particular valve not only depends on starting of the control line with particular valve fluid communication, also takes The state of the separator valve certainly started in independent control line.Therefore, the order of pressurization determines panel The logic output of layout.Therefore, some embodiments of the present invention provide employing conventional microfluidic dress Put the most obtainable device layout and function.

According to one embodiment of the invention, it is provided that to the microfluidic device with multiple control line The method being configured.Serial or asynchronous logic is implemented according to the method.Described method includes starting First control line and valve is placed in the first state.Valve (can be in multiple valve) can respond to Described first starting of control line and be placed in off.Such as, separator valve 124 can respond to boundary The applying pressure and the pressurization of control line 120 and close of face accumulator 104.As in this specification In be described more fully and described more particularly below, store owing to separator valve is placed in valve 128 and accommodating Between long-pending device 102, separator valve can provide multiple control function, and this provides serial logic operation.

Described method also includes, after valve is placed in the first state, starts and is operable to valve Group is placed in the second control line of the second state.Valve is in the situation of the first state (i.e. cutting out) and stops valve Group is placed in the second state (such as, closed mode).As it is shown in figure 1, starting control line 110 Before close separator valve 124 stop valve 128 to respond to starting of control line 110 and close (with close Lock).Therefore, some embodiments of the present invention provide serial logic function, and wherein valve starts Order creates the different outcomes selected from several end-state.In this example, control line 110 rise Begin to start closedown and the locking causing valve 128.On the other hand, initial the starting of control line 120 causes The closedown of separator valve 124, its closedown stoping valve 128 and locking when being maintained at closed mode.To the greatest extent Pipe Fig. 1 shows and applies, to independent pressure source 102 and 104, control line 110 He that pressure is started 120, but other start method and comprise within the scope of the present invention.

According to another embodiment of the invention, it is provided that use microfluidic device to perform serial logic Another kind of method.The method being configured the microfluidic device with multiple control line includes passing through Start the first control line and set up the first state of microfluidic device, then start the second control line.Miniflow First state of body device can include that responding to starting of a plurality of control line is set on valve or closes Closed state.Such as, as it has been described above, started control line 120 before starting control line 110 and cause point Closedown every valve 124.Therefore, valve 128 does not respond to starting of control line 110 and locking is closed.

Described method also includes the second state setting up microfluidic device by starting the second control line, and After start described first control line.The programmable features of microfluidic device as herein described provides and depends on The different end-state of the microfluidic device of order are started in valve.Therefore, in this example, starting Start control line 110 before control line 120 and cause the closedown of valve 128 and locking and separator valve thereafter The closedown of 124.It is adiaphorous for closing separator valve after locking-valve 128, because check-valves 112 is protected Hold the constant pressure of the control line to check valve downstream, stop the closedown of separator valve to affect microfluidic device State.

Implement the example of serial logic as microfluidic device, the first state of microfluidic device can comprise Close or the first input line group (input line 142 that such as valve 126 is blocked) of blocked state and with Second input line group (such as input line of multiple reative cells fluid communication in described microfluidic device 132).Described second state can comprise and the institute of multiple reative cells fluid communication in described microfluidic device State the first input line group (such as input line 142), and close or the second input line of blocked state Group (input line 132 that such as valve 128 is blocked).Second state also can comprise the valve of closed mode Group (such as valve 118a-118e), thus stop fluid through connecting the manifold flow of the second input line group.

It is noted that while separator valve is started by control line 120, other embodiments can use another kind of by The separator valve that control line 110 starts, using the replacement or supplementary as shown separator valve.Therefore, example As, overhead route can from accommodate accumulator 102 lead to be positioned at interface accumulator 104 and check-valves 127 it Between extra separation valve.Pressure is applied at the section 125 to the control line being connected to interface accumulator 104 Close before power this extra separator valve valve 126 can be stoped to respond to starting of control line 120 and close and Locking.Therefore, while figure 1 show that be embodied as, the invention is not restricted to this and be embodied as, other Microfluidic device designs within the scope of the present invention.

Although additionally, the embodiment shown in Fig. 1 and replaceability geometric shape discussed above have employed The separator valve that started by the row replacement plus-pressure in two control lines and provide multiple Two levels controlled, embodiment of the present invention provide other levels of multiple control.Such as, Other separate valves (such as, electrostatic starts) can be integrated in the design of microfluidic device, with The control of the 3rd level is provided.In addition to the starting of 2 shown control lines, the independence other to this The initial logic output starting meeting driving means state of valve.

Fig. 7 shows what the programmable microfluidic device to one embodiment of the invention was programmed Method.In the embodiment depicted in fig. 7, described microfluidic device is programmed for described microfluid The reative cell of device is separated into 48 independent panels.The specific number of panel depends on that microfluid fills The actual design put, and can be different numbers in the embodiment of replaceability, such as 12 or 192 Individual panel.The reative cell of each separate panels can fill different samples, provides in the present embodiment Detect while up to 48 different samples.

As shown in fig. 1, microfluidic device has the reaction site or reaction being arranged in array 106 Room array.Described reaction site is in fluid communication with the first input line group 132 and the second input line group 142. Described method includes that starting (710) is operable to block the first of (712) first input line groups Valve group.Such as, the first valve group is valve 128, is also referred to as operable to stop or close inlet flow The receiving valve of body line 132.Such as by applying pressure closes valve 128 and hinder accommodating accumulator 102 Fluid stopping body is flowed by input line 132.Owing to unidirectional or check-valves 112 existence makes control line 110 Becoming lock-in control line, after accommodating valve 128 closedown, they remain turned-off.

Described method also includes that starting (714) is operable to separate (716) described reaction site The second valve group.As shown in Figure 6B, reative cell 630 is limited at the elastomer layer of microfluidic device In 603.Described second valve group is arranged in array, and is shown as valve 615 in fig. 6.Except Outside one valve group 128 and the second valve group 615, the 3rd valve group 118a-118e is with described first valve group simultaneously Start.The existence of check-valves 112 also makes valve 118a-118e i.e. locking after closedown.Shown in Fig. 1 Embodiment in, described first valve group and described second valve group respond to control line 110 apply Pressure starts simultaneously or closes.

After described first, second, and third valve group is closed, described second valve group is cancelled and is started (718), Such as, by reducing or eliminating pressure control line 110 applied by pressure source 102.Described valve Cancelling and start the unlatching causing described valve, this allows fluid flow the input line being connected with valve.Multiple samples It is loaded into reaction site (720) by the second input line group 142.In FIG, sample is included in sample In input or mouth 140.The both sides of described microfluidic device respectively comprise 24 different input ports, carry Totally 48 input ports are supplied, 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 multiple input ports, can Comprise the sample that sum is less.Therefore, although table 1 relates to 48 panel layouts, programmable microfluidic Device provides up to 48 independent panels indeed according to the sample provided at 48 sample ports.? After sample is loaded into reaction site, second time starts the second valve group so that reative cell separate (722).

It should be understood that the concrete steps shown in Fig. 7 provide the microfluid to one embodiment of the invention A kind of concrete grammar of device programming.Other sequence of steps real also dependent on alternate embodiment Execute.Such as, the alternate embodiment of the present invention can implement step listed above in a different order Suddenly.Additionally, single step shown in Fig. 7 can comprise many sub-steps, they can be according to single step with many Plant order to implement.Additionally, can add according to concrete application or remove other step.This area is general Lead to and skilled person will comprehend that many changes, improves and replace.

Fig. 8 shows that the programmable microfluidic device to another embodiment of the invention is programmed Method.The method that Fig. 8 shows provides having single panel and all reative cells comprise identical The method that the programmable microfluidic device of sample carries out operating.As it is shown in figure 1, described miniflow able to programme Body device has the reaction site array being in fluid communication with the first input line group and the second input line group.? In exemplary shown in Fig. 8, described first input line group is to be connected with sample input 140 Input fluid line 142, described second input line group is and measures the input fluids that are connected of input 130 Line 132.First input line group is started for closed position (810), block through the first input line group Flowing (i.e. closing) (812).Therefore, in an example, interface accumulator 104 pressurization is led Cause the closedown of valve 126, stop fluid to flow through input line 142.The existence of check-valves 127 causes One valve group is latched in closed position.

Described method also includes that starting (814) is operable to block (816) second input line groups Second valve group of the Part I of subgroup.Seeing Fig. 1, the Part I of described second input line group is The part of the input line 132 under valve 134, described second valve group is valve 134, operable its with resistance Plug or closedown and 6 mensuration input 5 in 130 6 relevant input lines 132.Therefore, exist In the present embodiment, the subgroup of input line is 5 in 6 input lines, uses shown in Fig. 1 The mensuration input port (being connected with the input line of low order end) of right-hand member provides and is loaded into reative cell or the sample in site Product.The input line (i.e. the input line of low order end) being not included in subgroup is opened and is used for such as As being hereafter described more fully, sample is loaded into reative cell.Other are not used to survey in this is embodied as Determine input port.In the embodiment depicted in fig. 1, the pressure that interface accumulator 104 applies is opened simultaneously Dynamic valve 126 and valve 134.

See Fig. 1, the starting of valve that use by control line 110 carried out is not carried out.Therefore, will Sample is loaded into the mensuration input port of low order end and sample can be caused to flow through section 116 and flowed by manifold 135 Part II (i.e. the part of the input line 132 on valve 134) to described second input line group. Sample is loaded in the reative cell in array 106 or site (818) by described second input line group.Should Note by starting so that discrimination through the section 116 of the most uninflated control line 110 Pipe disables (i.e. the flowing through manifold is prevented from).Therefore, manifold is opened and in input line 132 Part II preliminary sectors connect all 6 input fluid lines.Through manifold 135, sample meeting It is assigned to the component of input line 132, finally flows through 24 input lines, through the valve opened 128.Therefore, all reative cells single sample being supplied in array 106.In order to separate reaction Sample in room (822), starts the 3rd valve group (820), such as the receiving valve 615 shown in Fig. 6 A. Can provide in the closed array 106 of pass by using receiving accumulator 102 that control line 110 applies pressure The pressure of valve 615.

It should be understood that the concrete steps shown in Fig. 8 provide the miniflow to another embodiment of the invention A kind of concrete grammar that body device is programmed.Other sequence of steps is also dependent on replaceability embodiment party Case is implemented.Such as, the alternate embodiment of the present invention can be implemented listed above in a different order Step.Additionally, single step shown in Fig. 8 can comprise many sub-steps, it can be according to single step Implement with multiple order.Additionally, can add according to concrete application or remove other step.Ability Territory those of ordinary skill is it will be appreciated that many changes, improves and replace.

Fig. 9 shows the letter being programmed of the microfluidic device to further embodiment of the present invention Change method.Described programmable microfluidic device has and the first input line group, the second input line group and company Connect the reaction site array of the manifold fluid connection of described second input line group.Described method includes starting (910) it is operable to block the first valve group of (912) first input line groups.See Fig. 1, First valve group can be the valve 126 in check-valves 127 downstream.The existence of check-valves 127 makes valve 126 open Closed position it is latched in after Dong.Start the second valve group (914) to block the second input line group subgroup Part I (916).Control line 120 is applied the pressure from interface accumulator 104 and can cause valve 126 and the starting of valve 134.

Seeing Fig. 1, the Part I of described second input line group is the input line 132 under valve 134 Point, the second valve group is valve 134, and it is operable to block or close measure input 130 with 6 In 6 input lines connected 5.Therefore, in the present embodiment, the subgroup of input line is 6 In bar input line 5.Because valve 134 is not by unidirectional valve blocking, when reopening these valves, Fluid can flow through the Part I of all 6 input lines 132 and flow to manifold 135.Shown in Fig. 1 In embodiment, apply the pressure from interface accumulator 104 and start valve 126 and valve 132 simultaneously.

Described method also includes starting the 3rd that (918) are operable to make manifold disable (920) Valve group.In an exemplary embodiment, the 3rd valve group start include to the second valve group fluid The pressure source of connection applies predetermined pressure at (such as, accommodating accumulator 102).See Fig. 1, logical Cross the section 116 to control line 110 to apply pressure and start valve 118a-118e and can block through manifold Flowing, to provide the separation to the fluid flowing through input line 132.The existence of check-valves 112 can cause Valve 118a-118e is latched in closed position.As described more completely below, disabling of manifold 135 makes 12 samples (6, the every side of microfluidic device sample) can measure input port 130 from 6 and be loaded into Reative cell.As discussed in the embodiment of Fig. 7, by identical sample is supplied to more than one Individual mensuration input can be loaded into less than 12 samples.It will be appreciated by those skilled in the art that many changes, Improve and replace.

Referring back to Fig. 1, it should be noted that before starting control line 110, the initial of control line 120 starts not Only result in valve 126 and the closedown of valve 134 and locking, also result in the closedown of separator valve 124.Owing to dividing Being arranged between receiving accumulator 102 and valve 128 every valve, the closedown of separator valve can stop valve 128 to be answered Answer and start in control line 110 and close.In the embodiment depicted in fig. 9, it is used for starting control line The pressure of 120 is predetermined pressure, and it be enough to stop opening of the control line 110 causing valve 128 to be closed Dynamic.As shown in table 1, the pressure of 45psi be enough to when control line 110 applies the pressure of 30psi Valve 128 is maintained at open position.Concrete value 45psi and 30psi are not essential to the invention, Other pressure can be used to implement this multiple control function.

Described method also includes cancelling starts the second valve group (922).Generally, by applying to first First pressure reduction of pressure source is such as cancelled to 0psi and is started the second valve group, so that it is taked out Open position.The reduction of the pressure being applied to control line makes the flexible membrane being deflected into described input fluid line return Going back to the position not deflected, the fluid flowing flowing through input fluid line recovers.Reopening valve After 134, the sample provided in measuring input 130 is written into reaction site (924).Such as Fig. 1 institute Show, Part I (the such as portion of the input line 134 under valve 134 that the second input line group is each Point) be in fluid communication with mensuration input port, described mensuration input port is arranged to receive one of multiple sample. What sample flowed through the second input line group first and second is partially into reaction site.Such as Fig. 6 A and 6B Shown in, described reaction site can be formed with the form of multiple reative cells, and the most each reative cell is by logical Hole is in fluid communication with the Part II of input fluid line 132, and described through hole arrives through input fluid line Described reative cell, described reative cell can be disposed at the elastomer microfluid dress on the layer containing fluid line Put in layer.

As it is shown in figure 1, the element branches being positioned at the input line 132 on valve 134 becomes four input lines Group, itself and the reaction site in array 106 or room fluid communication.By in each mensuration input port 6 different samples are provided, can define in an array altogether 12 panels (owing to Fig. 1 illustrate only First side of microfluidic device).As apparent to those skilled in the art, more than one Input port use a sample can result in less than 12 panels.In order to separate reaction site, Start the 4th valve group (926), such as valve 615.

In some implementations, described method also includes the closedown starting valve to stop the 5th valve group, wherein Operable 5th valve group is to close the Part II of the second input line group.As it is shown in figure 1, separator valve 124 close stops valve 128 to respond to starting and closing of control line 110.In the reality shown in Fig. 9 Executing in scheme, the pressure of the 45psi being applied to control line 120 time initial causes the closedown of separator valve, The Part II making input line 132 is held open manifold simultaneously and is stopped by the closedown of valve 118a-118e With.Separator valve can start by control line is applied pressure or can otherwise start, including But it is not limited to machinery, electrostatic etc..In the application using control line based on pressure, can use single Control line starts multiple valve and/or valve group simultaneously.Therefore, when interface accumulator 104 is pressurizeed, Separator valve 124 and valve 126 and valve 134 respond to starting and actuateding of control line 120 simultaneously.

The programmable nature of microfluidic device provided herein is proved by following facts: changes valve and starts step Rapid order can cause the valve in array to be in different states, and thus makes array be in different shapes State.Such as, in the embodiment depicted in fig. 9, by following steps, 12 samples can be loaded into Array, described step is: closing separator valve 124, then control line 110 is applied pressure, this makes Manifold 135 disables, and then reduces the pressure to accumulator 102, then reduces accumulator 104 Pressure.In this example, the pressure to accumulator 104 is kept by accumulator 102 when pressure is applied Power with the pressure that stops valve 128 to respond to use accumulator 102 to apply the control line 110 that causes it The closedown started.On the other hand, when applying pressure by accumulator 102, release is applied to accumulator The pressure of 104 can cause the closedown of valve 128.Therefore, the order started by changing valve achieves micro- Two different conditions of fluid means, are loaded into sample by input line 132 or close input line 132 Ability.This programmable features provides the employing the most obtainable benefit of conventional design.

It should be understood that the concrete steps shown in Fig. 9 provide programs the micro-of further embodiment of the present invention A kind of concrete grammar of fluid means.Other sequence of steps is also dependent on some alternate embodiments Implement.Such as, some alternate embodiments of the present invention can be implemented in a different order with above-listed The step gone out.Additionally, single step shown in Fig. 9 can comprise many sub-steps, it can be according to single step Suddenly implement with multiple order.Additionally, can add according to concrete application or remove other step.This Field those of ordinary skill is it will be appreciated that many changes, improves and replace.

It should be understood that the invention is not restricted to concrete grammar as herein described, scheme and reagent etc., this area skill Art personnel will understand that these change.Will also be understood that term used herein is intended merely to for describing tool The embodiment of body, it is no intended to limit the scope of the present invention.Should also be noted that as in this article with appended Used by claim, unless the context clearly indicates, when not indicating quantity, include plural number Form.It is therefoie, for example, mention one or more cell, this area skill when mentioning " cell " Art personnel its equivalent known.

Except as otherwise noted, to have the technical field of the invention general for all scientific and technical terminologies used herein The same meaning that logical technical staff is generally understood that.Embodiment of the present invention and various features thereof and advantage Details describe in detail and/or the non-limiting embodiments of explaination with reference in accompanying drawing and description below It is explained more fully with embodiment.It should be noted that feature shown in accompanying drawing be not necessarily according to than Example is drawn, and as understood by a person skilled in the art, even if not explicitly pointing out in this article, one Feature in individual embodiment can be used for other embodiments.To known assemblies and the description for the treatment of technology Can be omitted to avoid and embodiment of the present invention is produced unnecessary obscuring.Example used herein is only It is for the ease of understanding the mode that the present invention can be carried out, and makes those skilled in the art implement further Embodiment of the present invention.Therefore, example and embodiment herein is understood not to limit this Bright scope, the scope of the invention is only limited by claims and applicable law.Furthermore, it should be noted that Reference similar in several views of accompanying drawing represents similar part.

Therefore, following present " definition " part, wherein in order to clearly specifically define and the present invention Some relevant terms, but it is defined and those skilled in the art's understanding one to these terms Cause.Describe concrete method, equipment and material, although or equivalent similar with those described herein Any method and material can be used for implement or test the present invention.The all lists of references that present document relates to It is incorporated herein by reference by entirety.

Definition

PNA is peptide nucleic acid(PNA) (peptide nucleic acid)

LNA is lock nucleic acid (locked nucleic acid)

DA is dynamic array (dynamic array)

PCR is polymerase chain reaction (polymerase chain reaction)

BSA is bovine serum albumin (bovine serum albumin)

FRET is FRET (fluorescence resonance energy transfer) (Fluorescence resonance energy transfer)

GT is gene type (genotyping)

PEG is Polyethylene Glycol (polyethylene glycol)

PLP is padlock probe (padlock probe)

Terms used herein " neighbouring " typically refers to primer and visits relative on target nucleic acid analyte complementary strand The position of pin.Primer and probe can be spaced about 1 to about 20 nucleotide, more specifically, about 1 to about 10 nucleotide, or can be immediately adjacent to one another.

Terms used herein " analyte " typically refer to the inventive method to be used detection or quantitative with Undefined nucleic acid molecules or nucleic acid molecules mixture.Term " target nucleic acid analyte " and " nucleic acid divides Analysis thing " use can be exchanged with term " analyte " in the present invention.

Terms used herein " complementary " or " complementary " can be included under salt and the temperature conditions of permission The polynucleotide natural combination by base pairing.Such as, sequence " A-G-T " combines its complementary sequence Row " T-C-A ".Complementation between two single chain molecules can be " part ", the most only one A little nucleic acid combine, or the complementation between two single chain molecules exists overall complementary between single chain molecule It is completely during property.Hybridization efficiency between nucleic acid chains and intensity are had by the complementarity between nucleic acid chains Important function.This is in the amplified reaction of the design and use of the combination and molecule of depending on nucleic acid interchain It is even more important.

Terms used herein " covalently bound " typically refer to a molecular structure by covalent chemical bond with The connection of another molecular structure.

Terms used herein " dyestuff " typically refers to absorb electricity when wavelength is more than or equal to 340nm Any organic or inorganic molecule of magnetic radiation.

Terms used herein " fluorescent dye " typically refers to by electromagnetic radiation source (such as, lamp, light Electric diode or laser) irradiate after sent any dye of electromagnetic radiation of longer wavelength by Fluorescence Mechanism Material.

Terms used herein " GT sample buffer " typically refers to can be by reaction channel in DA chip The buffer closed with the binding site of chamber surface.Described buffer is when chip loading processes or reacts Prevent the loss of reactive component.For reagent cost, other Taq-Gold also can be gathered by it The use of synthase reduces less than about 80%.20 × GT buffer can include following combination: glycine betaine (FW 117.15)、BSA、T20 (in PBS) (Thermo Scientific, Rockford, IL),(in PBS) (Thermo Scientific, Rockford, IL)、(in TBS) (Thermo Scientific, Rockford, IL),T20 (in TBS) (Thermo Scientific, Rockford, IL), glycerol, PEG 20,000, PEG MME550, PEG MME5000 and Tween 20.

Terms used herein " homogeneity measures (homogenous assay) " typically refers to detection or fixed The method of amount nucleic acid analyte, it is without measuring post processing to record measurement result.Homogeneity measures can be The pipe closed or microfluidic arrays are carried out, and need not add further reagent after mensuration is initial Or supplementary chemicals are to record result.Homogeneity measures the result that energy real time record measures, it means that survey Determine result to be recorded continuously when measuring progress in time.

Terms used herein " hydrolysis probes " has carried out generality in United States Patent (USP) No.5,210,015 Describing, it is incorporated herein by reference by entirety.Hydrolysis probes make use of used by PCR reacts 5 '-nuclease present in heat stability Taq polymerase (Probe technique, Applied Biosystems, Foster City CA).With fluorescent detection dye (such as fluorescein) with connect Receive dyestuff or quencher labelling hydrolysis probes.Generally, fluorescent dye is covalently attached to 5 ' ends of probe, Quencher is connected to 3 ' groups of probe, and when probe is complete, the fluorescence of detection dyestuff is due to fluorescence altogether The energy that shakes shifts (FRET) and is quenched.Probe can at the anneals downstream of a primer, described in draw Thing limits an end of PCR reaction nucleic acid target analyte amplification target position.Utilize the polymerization of Taq enzyme Enzymatic activity, the amplification of target nucleic acid analyte by a primer of probe upstream and be positioned at probe downstream but On the relative chain of target nucleic acid, the second primer of annealing guides.Along with the extension of forward primer, Taq is polymerized Enzyme arrives the region of label probe annealing, probe-template crossbred is identified as substrate, and hydrolyzes spy The phosphodiester bond of pin.Hydrolysis irreversibly discharges quencher dyes and makees the cancellation of reporting dyes With, thus result in along with each continuous print PCR cycle, detector fluorescence increases.Specifically, The hydrolysis probes of the present invention can be at common 8-mer or 9-mer motif in people and other transcript profile can By the high T using LNA analog to have about 70 DEG C_m。

Terms used herein " labelling " refers to can be used for provide and can detect and/or can the appointing of quantifiable signal What atom or molecule.Specifically, described labelling is connectable to nucleic acid or protein.Labelling can provide The detections such as fluorescence, radioactivity, colorimetric, X-ray diffraction or absorption, magnetic, enzymatic activity can be passed through Signal.

Terms used herein " nucleic acid " typically refers to cDNA, DNA, RNA, its strand or double Chain and any chemical modification, such as PNA and LNA.LNA United States Patent (USP) No.6,794,499, 6,670,461,6,262,490 and 6, described in 770,748, and it is expressly incorporated herein by quoting in full.Core Acid can be any size.Nucleic acid is modified and can be included adding chemical group, its to single nucleic acid base or Nucleic acid entirety adds extra electric charge, polarity, hydrogen bond, electrostatic interaction and function.This modification Can include that the base such as 2 '-position modified is sugar-modified, 5-position pyrimidine is modified, 8-position purine is modified, cytosine The modification of exocyclic amine, the replacement of 5-bromouracil, backbone modification, methylate, unusual base pairing Combine cytidine as different in different base and different guanidine etc..Nucleic acid may be from chemistry building-up process completely (such as solid phase The chemosynthesis of mediation) or come from biogenetic derivation (as by from the substantially any thing that can provide nucleic acid Kind separates) or come from process (the such as DNA related to by biology tool operation nucleic acid Replicate, PCR amplification, reverse transcription) or from the combination of these processes.

Terms used herein " nucleic probe " is to carry at least one covalently bound dyestuff (as fluorescence contaminates Material) nucleic acid.Specifically, probe does not comprise and the complementary for causing PCR to react Sequence.

Terms used herein " padlock probe " or " PLP " typically refer to have about 100 base pairs The linear oligonucleotide of length.The sequence of PLP 3 ' and 5 ' end and the neighbouring sequence in target nucleic acid analyte Row complementation.There is " the label sequence that can be used for identifying specific PLP in the central incomplementarity region of PLP Row ".The flank of described sequence label can be universal primer site or unique and/or specific primer position Point, it allows PCR to expand described sequence label.After being hybridized to target, the 5 ' and 3 ' of PLP End is the most adjacent and can be connected subsequently.Products therefrom is the cycling probe connecting target nucleic acid analyte Molecule.Such as can useIn real time PCR to the label area amplification of cyclisation PLP and Quantitatively and/or detection.The existence of amplicon and amount can be associated with existence and the amount of target sequence in sample. Description to PLP see 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 by full It is incorporated herein by reference.

Terms used herein " PCR " typically refers to for expanding, detect or quantifying analytes given zone The method in territory.It will be understood by those skilled in the art that in basic PC R technical foundation, have several change, Such as allele specific pcr, set PCR (assembly PCR) or polymerase cycle set (polymerase cycling assembly, PCA), colony PCR, helicase dependent amplification, Specificity (intersequence-specific, ISSR) PCR between heat start PCR, sequence, reversely PCR, ligation-mediated PCR, methylation status of PTEN promoter, many sites join dependency probe amplification, Multiplex PCR, nest-type PRC, Overlap extension PCR, quantitative PCR, quantitatively in real time PCR, RT-PCR, hot asymmetric interlaced (thermal asymmetric interlaces, TAIL) PCR, Touchdown PCR and PAN-AC.How in addition, those skilled in the art will appreciate that should by these changes It is used on the round pcr on basis.

Terms used herein " purification " typically refers to will according to electric charge, molecular size or binding affinity Any process that protein, polypeptide or nucleic acid separate with other compositions or compound.

Terms used herein " quencher " typically refers to reduce the dyestuff of the fluorescent emission of another dyestuff.

Terms used herein " inquiry (querying) " typically refers to whether determine target specific probe Relevant to nucleic acid analyte (such as, connect or link), and optionally quantitatively in sample target special The amount of probe.

" sample " used herein typically refers to the tissue from human or animal or fluid sample, including but It is not limited to blood plasma, serum, spinal fluid, lymph fluid, skin, respiratory system, intestinal and urogenital tract Outer section, tear, saliva, hemocyte, tumor, organ, tissue and Cell culture invitro group The sample divided.Specifically, sample can be unicellular, paraffin-embedded tissues and the work of acupuncture biology Sample product.Additionally, sample can include environmental sample, such as lake water and foodstuff samples.

It is natural that word used herein " basic purification " or " being basically separated " generally include from them Environment obtains, separate or the nucleic acid that separates or aminoacid sequence, they at least without about 60%, special Other components associated there of not at least about 75%, the most at least about 90%, and Separate nucleic acid thing including restructuring or clone is similar with what chemosynthesis analog or system biological synthesized Thing.

In view of polymer chemistry, precursor, synthetic method, reaction condition and available additive Greatly multiformity, has substantial amounts of useful elastomers system to can be used for manufacturing elastic module, layer, film, micro- Type valve, pump etc..The change of material therefor in some cases can due to the specific material behavior of needs, I.e. solvent resistance, rigidity, breathability or temperature stability determines.There is the most eurypalynous flexible polymer Thing.Provide being briefly described of modal elastomer type herein, it is therefore an objective to even if showing to use relatively The polymer of " standard ", there is also the multiple possibility of combination.Conventional elastomeric polymer includes poly-isoamyl Diene, polybutadiene, polychlorobutadiene, polyisobutylene, poly-(s-B-S), Polyurethane and silicone or polysiloxanes.

Polyisoprene, polybutadiene and polychlorobutadiene are polymerized by diene monomers, therefore exist During polymerization, each monomer has a double bond.This double bond makes polymer pass through sulfuration, and (generally, sulfur is used for Between double bond, crosslinking is formed by heating) it is converted into elastomer.This makes by treating binder course Incomplete vulcanization carries out the easy soft printing of homogenizing multilamellar (homogeneous multilayer soft lithography)；Photoresist encapsulation (photoresist can be carried out by similar mechanism encapsulation)。

Pure polyisobutylene does not have double bond, but by introducing the different of a small amount of (about 1%) when polymerization Pentadiene carries out cross-linking thus is used as elastomer.This isoprene monomer is given on polyisobutylene skeleton Pendency double bond, it can cure the most as described above.

Poly-(s-B-S) passes through active anionic polymerization (the most in the reaction without natural Chain termination step) produce, therefore can there is " active " polymer ends at the polymerization species of solidification. This is that the natural material standed for of photoresist package system is (wherein at the liquid layer being cast in cured layer top In have a large amount of unreacted monomer).Incomplete solidification allows for the soft printing of homogeneous multilamellar (A with A is combined).This chemical property be additionally favorable for making one layer have extra butadiene (" A ") and Coupling agent, another layer (" B ") lacks butadiene (for the soft printing of homogeneous multilamellar).SBS is " heat Solidity elastomer ", mean its fusing plastic (relative with elasticity) on uniform temperature；Reduce temperature Degree produces elastomer again.Therefore, layer can be combined by heating.

Polyurethane is made up of diisocyanate (A--A) and glycol or diamidogen (B--B)；Due to two Isocyanates and glycol/amine have a variety of, the enormous amount of different types of polyurethane.But, poly- The A of the compound characteristic relative to B makes them for the soft printing of heterogenous multilayer as RTV 615: By using the A-A of excess in one layer, another layer uses the B-B of excess.

Siloxane polymer has the change of great structure, and provides multiple commercial preparation.Discuss The vinyl of RTV 615-(the soft printing of heterogenous multilayer and photoresist can be carried out to (Si--H) crosslinking Encapsulation), but this is only intended to one of several cross-linking methods of siloxane polymer chemistry.

In addition to using above-mentioned simple " pure " polymer, cross-linking agent can be added.Some reagent (as There is the monomer of pendency double bond for sulfuration) be suitable for the soft printing of homogenizing (A to A) multilamellar or Photoresist encapsulates；Identical reagent is mixed in the method in two elastic layers.Complementary reagents is (i.e. A kind of monomer has pendency double bond, and another kind has pendency Si--H) it is suitable for heterogeneous (A To B) the soft printing of multilamellar.Mutual tonic is added in the method to adjacent layer.

Below for the non-exhaustive list of elastomer can being used in combination with the present invention: polyisoprene, poly- Butadiene, polychlorobutadiene, polyisobutylene, poly-(s-B-S), polyurethane and Siloxane polymer；Or poly-(two (fluoroalkyl) phosphine nitrile) (PNF, Eypel-F), poly-(carborane- Siloxanes) (Dexsil), poly-(acrylonitrile-butadiene) (nitrile rubber), poly-(1-butylene), poly-(chloro three Fluorothene-difluoroethylene) copolymer (KeI-F), poly-(ethyl vinyl ether), poly-(difluoroethylene), poly-(two Fluorothene-hexafluoropropene) copolymer (Viton), polrvinyl chloride elastic composition (PVC), polysulfones, poly- Carbonic ester, polymethyl methacrylate (PMMA) and politef (Teflon).

Allcock etc., Contemporary Polymer Chemistry, elastomer is retouched by the second edition State as the polymer at a temperature of being present between glass transition temperature and condensing temperature.Elastomer display Elastic characteristic is owing to the motion that is prone to twist of, polymer chain makes skeletal chain solve under force Opening, when unable effect, skeletal chain curling returns shape originally.Generally, elastic when force is applied Body deformability, reverts to original shape when removal force.The elasticity of elastomer display can use modulus of elasticity table Levy.The modulus of elasticity of the material that some embodiments of the present invention use be about 1Pa to about 1TPa, Or about 10Pa to about 100GPa or about 20Pa to about 1GPa or about 50Pa to about 10MPa, Or about 100Pa to about 1MPa, although also can use outside these scopes according to the needs of concrete application Modulus of elasticity.In some cases, the modulus of elasticity of material can be about 100MPA (MPa) Or it is less.In other embodiments, the modulus of elasticity of material is about 75MPA or less, about 50MPA or less, about 25MPA or less, about 10MPA or less, about 8MPA or more Less, about 5MPA or less or about 2MPA or less.

Embodiment of the present invention provide microfluidic device, and it comprises assembly such as passage, valve and room, Described assembly is at least partly comprised by fast one or more layer of springform or level, embeds, or Person is formed by it or is formed wherein.A kind of exemplary microfluidic body device has and is formed at elastomer first Reagent flow passage in Ceng or reagent line.Described reagent flow passage comprises receiving valve and room conduit.Miniflow Body device also can have the control being formed in the elastomer second layer adjoining with described ground floor and lead to Road or receiving line.Additionally, microfluidic device can comprise is formed at the elasticity adjoining with the described second layer Sample flow channel in body third layer or sample wire.Described sample flow channel can comprise receiving valve and room is led Pipe.Described control passage can accommodate valve with reagent flow passage and sample flow channel accommodates valve operability even Connect.Described microfluidic device can comprise and reagent line fluid communication reagent chamber, and with sample wire fluid The sample room of connection.Described reagent chamber and sample room can be by being formed in described elastomer third layer Reaction stream passage or response line and fluid communication with each other.Described response line can comprise interface valve.Described micro- Fluid means also can comprise the interface being formed in the 4th layer of the elastomer adjoining with described third layer Passage.Described interface channel can be connected with described reaction stream Entry Interface valve operability.

The method that embodiment of the present invention are also contemplated by manufacturing and using microfluidic device disclosed herein. Such as, the operation of microfluidic device can include opening one or more separate valves, close one or more Multiple interface valves and make material flow through separate valves and flow into one or more room, the most under stress Carry out.Technology may also include the pressure changed in seal line to close separate valves, to close separate chambers, And change the pressure of parting line to open interface valve.First material of the first Room can flow through the boundary of unlatching Face valve enters the second Room, wherein the first material and the second material mixing contained therein or reaction.

It should be understood that example as herein described and embodiment are intended merely to example purpose, people in the art Member can advise multiple improvement or change according to it, and these are included in spirit herein and authority And in scope of the following claims.

Herein below is corresponding to the original claims in parent application, now as one of description Divide and be incorporated herein by:

1, a kind of microfluidic device, it comprises

Pressure source；

Control line, it is in fluid communication with described pressure source；

Multiple valves, it is operated by described control line；With

Separate valves, its be positioned near described control line and be in described pressure source and the plurality of valve it Between.

2, the microfluidic device of item 1, wherein said separate valves builds and is arranged in obstruction fluid and passes through The flowing of described control line.

3, the microfluidic device of item 1, wherein said separate valves and the second pressure source fluid communication.

4, the microfluidic device of item 1, wherein said control line includes the lock-in control comprising check valve Line.

5, the microfluidic device of item 4, wherein said check valve is arranged to stop fluid from described valve Flow to described pressure source.

6, the microfluidic device of item 5, it also comprises multiple rooms valve.

7, the microfluidic device of item 6, the wherein said the plurality of valve of control alignment and the plurality of room Valve provides and starts pressure.

8, the microfluidic device of item 1, wherein said separate valves can be used to stop the plurality of valve Close.

9, the microfluidic device of item 1, it also comprises more than second valve by the second control line operation.

10, the microfluidic device of item 9, wherein said second control line includes comprising the second check valve Second lock-in control line.

11, the method operating the microfluidic device with valve and control line, described control line has and it The valve group being associated, described method includes:

Close described valve；With

Described control line is applied pressure；The valve wherein cut out makes the institute being associated with described control line State valve group inoperable.

12, the method for item 11, wherein closes described valve and includes:

Second control line is applied the second pressure；With

Respond to described second pressure and close described valve.

13, the method for item 11, wherein the first valve group responds to and described first control line is applied first Pressure and close.

14, the method for item 13, wherein said first valve group is blocked,

15, the method for item 11, it also includes:

Keep applying described second pressure to described second control line；With

Described first control line is applied less than the second pressure of described first pressure, wherein with described the The valve group that two control lines are associated responds to the applying of described second pressure and closes.

16, the method for item 15, wherein said valve group is blocked.

17, a kind of microfluidic device, it comprises:

First valve；

Second valve；

Control line, itself and described first valve and described second valve fluid communication；

Acute build up of pressure device, it is in fluid communication with described control line；With

Check valve, it is positioned near described control line and is positioned at described acute build up of pressure device and described second valve Between.

18, the microfluidic device of item 17, between wherein said acute build up of pressure device and described first valve Described control line is without check valve.

19, the microfluidic device of item 17, wherein said first valve is arranged to described micro-by being arranged in Reative cell in fluid means separates.

20, the microfluidic device of item 17, wherein said second valve is arranged to input first fluid Line and second fluid input line separate.

21, the microfluidic device of item 17, it also comprises the 3rd valve being in fluid communication with described control line.

22, the microfluidic device of item 21, wherein said 3rd valve can be used to stop fluid to pass through The flowing of the fluid input line mutually coupled with reative cell.

23, the microfluidic device of item 21, it also comprises structure and is arranged to block fluid through described control Line processed flowing separator valve, wherein said separator valve along described control line be placed in described acute build up of pressure device and Between described 3rd valve.

24, the microfluidic device of item 23, wherein said separator valve is set to accept to control from second Start pressure, described second control line and the second acute build up of pressure device of line are in fluid communication.

25, the microfluidic device of item 24, wherein said second control line is independent of described control line.

26, the microfluidic device of item 23, wherein said separator valve can be used to stop the described 3rd Valve responds to described starting of control line and cuts out.

27, a kind of microfluidic device, it comprises:

Multiple reative cells, it is arranged as array layout, and each of the plurality of reative cell has with described First valve of one of multiple reative cells fluid communication and one of the described fluid with the plurality of reative cell Second valve of connection；

First control line, it is operable to start described first valve and described second valve；

Input line group, it is in fluid communication with the plurality of reative cell；

Multiple sample inlets, it is in fluid communication with described input line group；With

Check valve, it is arranged in described first control line.

28, the microfluidic device of item 27, its also comprise with first acute build up of pressure device fluid communication and can It is operated to block the first valve group of a part of described input line group.

29, the microfluidic device of item 27, wherein said first valve group is in the downstream of described check valve.

30, the microfluidic device of item 27, it also comprises:

Second input line group, it is in fluid communication with the plurality of reative cell；With

More than second sample inlet, itself and described second input line group fluid communication.

31, the microfluidic device of item 30, it also comprises can be by the second valve of the second control line operation Group.

32, the microfluidic device of item 31, wherein said second control line and the second acute build up of pressure device stream Body connects, and is operable to block a part of described second input line group.

33, the microfluidic device of item 31, it also comprises second be arranged in described second control line Check valve.

34, the microfluidic device of item 31, it also comprises and is arranged in described second control line and can quilt Operate in order to block the fluid separator valve through described first control line flowing.

35, a kind of microfluidic device, it comprises:

Multiple reative cells；

Multiple first input ports, each of wherein said multiple first input ports is inputted by more than first One of line and the one or more fluid communication in the plurality of reative cell；

Multiple second input ports, each of wherein said multiple second input ports is inputted by more than second One of line and the one or more fluid communication in the plurality of reative cell；

First acute build up of pressure device, itself and the first control line fluid communication, wherein said first control line sets It is set to close described more than first input line；

Second acute build up of pressure device, itself and the second control line fluid communication, wherein said second control line sets It is set to close described more than second input line；

First check valve, it is arranged in and is positioned at described first acute build up of pressure device and described more than first inputs In described first control line between line；With

Second check valve, it is arranged in and is positioned at described second acute build up of pressure device and described more than second inputs In described second control line between line.

36, the microfluidic device of item 35, it also comprises and is operable to separate the plurality of reaction Each valve group of room.

37, the microfluidic device of item 35, it also comprises and is operable to block described second and controls Line and be arranged in the valve between described second acute build up of pressure device and described second control line.

38, the microfluidic device of item 37, wherein said first control line can be used to start described Valve.

39, the microfluidic device of item 35, its also comprise can be used to block described more than second defeated Enter the valve group of the subgroup of line.

40, the microfluidic device of item 35, it also comprises the valve group that can be used to block manifold, institute State manifold and provide the fluid communication between described more than second input line.

41, the method operating the microfluidic device with multiple valve and check valve, described method includes:

First fluid pressure is applied to the control line of described microfluidic device；

Respond to the applying of described first pressure and close the plurality of valve；

Respond to the applying of described first pressure and close described check valve；With

Second fluid pressure is applied to the second control line of described microfluidic device.

42, the method for item 41, its also comprise reduce be applied in described control line described first-class Body pressure.

43, the method for item 42, the described first fluid being wherein applied in described control line in reduction After pressure, the plurality of valve remains off.

44, the method for item 42, wherein reduces described first fluid pressure and includes to described control line not Apply fluid pressure.

45, the method for item 41, wherein applies described second fluid press packet to described second control line Include closedown separator valve.

46, the method for item 45, wherein said separator valve can be used to stop the plurality of valve and institute State the fluid communication between control line.

47, the method for item 41, wherein said microfluidic device has multiple reative cell and with described The valve group that in multiple reative cells, each is associated.

48, the method operating the microfluidic device with multiple input port, described method includes:

Input fluid is provided to one of the plurality of input port；

Start the Part I of the input line that valve group is connected with the subgroup of the plurality of input port with closedown, Wherein said subgroup does not comprise the one of described of the plurality of input port；

Described input fluid is made to flow through the input line being connected with one of described in the plurality of input port；

Described input fluid is made to flow to the Part II of described input line through described input line；With

Close the second valve group to separate multiple reative cells.

49, the method for item 48, wherein closes described second valve group and includes the first pressure source is applied stream Body pressure.

50, the method for item 49, wherein closes described second valve group and includes closing between offer input line The manifold of fluid communication.

51, the method for item 49, wherein starts described valve group and includes the second pressure source is applied fluid pressure Power.

52, the method for item 48, wherein makes described input fluid flow to described the second of described input line Part includes that making described input fluid flow through provides the manifold of fluid communication between described input line.

53, the method for item 48, wherein started described valve group before closing described second valve group.

54, the method for item 48, it also includes making described input fluid flow through the second input line group, institute State the second input line group of branches and become the described Part II of each in described input line.

55, the method operating programmable microfluidic device, described microfluidic device has and the first input Line group and the reaction site array of the second input line group fluid communication, described method includes:

Start the first valve group being operable to block described first input line group；

Start the second valve group being operable to block the Part I of the subgroup of the second input line group；

By the Part II of described second input line group, sample is loaded into described reaction site；With

Start the 3rd valve group being operable to separate described reaction site.

56, the method for item 55, wherein starts described first valve group and includes the first valve group described in locking.

57, the method for item 55, wherein start described first valve group include to described first valve group stream First pressure source of body connection applies the first pressure.

58, the method for item 55, wherein starts described first valve group and starts described second valve group simultaneously Carry out.

59, the method for item 55, wherein start described 3rd valve group include to described 3rd valve group stream Second pressure source of body connection applies the second pressure.

60, the method for item 59, multiple samples are being loaded into institute by starting of wherein said 3rd valve group Carry out after stating reaction site.

61, the method for item 55, wherein said sample is through connecting the second of described second input line group The manifold of part flow to the Part II of described second input line group from the Part I of described input line, The Part I of described input line is not included in the subgroup of described second input line group.

62, the method for item 55, is wherein not included in the described subgroup of described second input line group Input line and the input port being set to receive described sample are in fluid communication.

63, a kind of microfluidic device, it comprises:

The input port of predetermined quantity, each described input port is operable to receive multiple input fluid One of；

Multiple input fluid lines, the input of each and described predetermined quantity of the plurality of input fluid line One of mouth fluid communication；

Valve group, each being operable to of described valve group close one of the plurality of input fluid line, The quantity of wherein said valve group is less than described predetermined quantity；

Manifold, it is in fluid communication with each of the plurality of input fluid line；With

Second valve group, each be operable to close described manifold one of described second valve group Point.

64, the microfluidic device of item 63, wherein said predetermined quantity is 12.

65, the microfluidic device of item 63, it also comprises and the first pressure of described valve group fluid communication Source.

66, the microfluidic device of item 65, it also comprises and the second pressure of described valve group fluid communication Source.

67, the microfluidic device of item 63, the described part of wherein said manifold comprises fluid line, institute State fluid line by one of the plurality of input fluid line with another of the plurality of input fluid line It is connected.

68, the microfluidic device of item 63, the quantity of wherein said valve group is less than described predetermined quantity Quantity.

69, the microfluidic device of item 63, wherein said multiple input fluid lines each is branched off into many Individual input fluid line.

70, the method operating programmable microfluidic device, described microfluidic device has reaction site battle array Row, itself and the first input line group, the second input line group and the manifold stream being connected described second input line group Body connects, and described method includes:

Start the first valve group being operable to close described first input line group；

Start the second valve group being operable to close the Part I of the subgroup of the second input line group；

Start the 3rd valve group being operable to make described manifold disable；

Cancel and start described second valve group；

By the Part II of described second input line group, multiple samples are loaded into described reaction site In；With

Start the 4th valve group being operable to separate described reaction site.

71, the method for item 70, wherein starts described first valve group and includes the first valve group described in locking.

72, the method for item 70, wherein starts described first valve group and starts described second valve group and include First pressure source is applied the first pressure, described first pressure source and described first valve group and described second Valve group is in fluid communication.

73, the method for item 72, wherein starts described 3rd valve group and includes the second pressure source is applied the Two pressure, described second pressure source and described second valve group fluid communication.

74, the method for item 73, wherein said second pressure is less than described first pressure.

75, the method for item 70, wherein cancels and starts described second valve group to include that reduction is applied to described Described first pressure of the first pressure source.

76, the method for item 70, wherein starts described 3rd valve group and includes the 3rd valve group described in locking.

77, the method for item 70, it also includes starting valve to stop the closedown of the 5th valve group, and described the Five valve groups are operable to close the described Part II of described second group of input line.

78, the method for item 77, wherein said valve and the first pressure source fluid communication, described first pressure Power source and described first valve group fluid communication.

79, the method for item 70, in wherein said second input line group, the Part I of each is with defeated Fluid communication, described input port is set to receive one of the plurality of sample.

80, the method for item 70, wherein said reaction site comprises multiple reative cell, the plurality of instead The each Part II by through hole with one of described second input fluid line group answering room is in fluid communication.

81, the method for item 70, in wherein said second input line group the Part I of each with not It is in fluid communication with input port.

82, the method operating programmable microfluidic device, described programmable microfluidic device has and the One input line group and the reaction site array of the second input line group fluid communication, described method includes:

Start the second valve group being operable to separate described reaction site；

Cancel and start described second valve group；

Multiple samples are assembled by the second input line and is loaded into described reaction site；With

Start described second valve group.

83, the method for item 82, wherein starts described first valve group and includes the first valve group described in locking.

84, the method for item 82, wherein starts described first valve group and includes the first pressure source is applied the One pressure, described first pressure source and described first valve group fluid communication.

85, the method for item 82, it is additionally included in while starting described first valve group and starts the 3rd valve Group.

86, the method for item 85, wherein starts described 3rd valve group and includes the 3rd valve group described in locking.

87, the method for item 82, wherein cancels and starts described first valve group to include that reduction is applied to described Described first pressure of the first pressure source.

88, the method for item 82, in wherein said second input line group, each flows from different input ports Body connects.

89, the method for item 82, wherein starts described first valve group and starts described second valve group simultaneously Carry out.

90, the method for item 82, wherein said reaction site includes being limited to described microfluidic device Reative cell in elastomer layer.

91, a kind of microfluidic device, it comprises:

Multiple reaction site；

First input line group, its provide predetermined quantity the first input port and the plurality of reaction site it Between fluid communication, the quantity of wherein said first group is described predetermined quantity；

Second input line group, it provides between the second input port and the plurality of reative cell of predetermined quantity Fluid communication, in wherein said second input line group, each comprises trunk portion and component, And the quantity of described second group is less than described predetermined quantity；With

Programmable input device, it is operable to use described first input line group or described second Input line group fills described reative cell.

92, the microfluidic device of item 91, wherein said multiple reaction site are arranged as array layout.

93, the microfluidic device of item 91, in wherein said multiple reaction site, each comprises reaction Room, described reative cell is by through hole and one of described first input fluid line group fluid communication.

94, the microfluidic device of item 91, in wherein said multiple reaction site, each comprises reaction Room, described reative cell is by through hole with one of the described second component inputting fluid line group fluid even Logical.

95, the microfluidic device of item 91, the described branch of each in wherein said input line group Divide and form four fluid passages from described trunk portion.

96, the microfluidic device of item 91, wherein said programmable input device comprises can be used by operation To close the valve group of described first input fluid line group.

97, the microfluidic device of item 96, wherein said valve group is in fluid communication with check valve, described list It is operable to described valve group is latched in closed mode to valve.

98, the microfluidic device of item 91, wherein said programmable input device comprises valve group, described Valve group is operable to close the described component of described second input fluid line group.

99, the microfluidic device of item 98, wherein said valve group is in fluid communication with check valve, described list It is operable to described valve group is latched in closed mode to valve.

100, the microfluidic device of item 98, it also comprises and is arranged near described valve group and can be grasped Act on the separate valves to stop described valve group to be closed.

The method that 101, the microfluidic device with multiple control line is set, described method includes:

Start the first control line；

Valve is placed in the first state；

Afterwards, start the second control line being operable to that valve group is placed in the second state, wherein locate Valve in described first state stops described valve group to be placed in described second state.

102, the method for item 101, wherein said first state is to close.

103, the method for item 101, wherein said second state is to close.

104, the method for item 101, wherein starts described first control line and includes executing the first pressure source Add the first pressure.

105, the method for item 104, wherein starts described second control line and includes executing the second pressure source Add the second pressure.

106, the method for item 105, wherein said second pressure source is independent of described first pressure source.

107, a kind of setting has the method for microfluidic device of multiple control line, and described method includes:

Then start the second control line set up described microfluidic device by starting the first control line First state；With

It is stream by starting described second control line and then start described first control line to set up described Second state of body device.

108, the method for item 107, wherein said first state includes that be closed first is defeated Enter line group and with described microfluidic device on arranged multiple reative cells fluid communication second input Line group.

109, the method for item 108, wherein said second state includes and institute on described microfluidic device Arrange the plurality of reative cell fluid communication described first input line group and be closed Described second input line group.

110, the method for item 108, wherein said second state also includes the valve group being closed, It stops fluid to flow through the manifold connecting described second input line group.

111, the method for item 107, wherein said first control line and the first pressure source fluid communication.

112, the method for item 111, wherein said second control line and the second pressure source fluid communication.

113, the method for item 107, wherein said microfluidic device comprises elastomeric material.

114, a kind of microfluid system, it comprises:

Carrier, it comprises:

Multiple first input ports；

Multiple first input lines, in the plurality of first input line, each is with the plurality of first One of input port is in fluid communication；

Multiple second input ports；

Multiple second input lines, in the plurality of second input line, each is with the plurality of second One of input port is in fluid communication；

First pressure source；With

Second pressure source

Dress microfluidic device on the carrier, described microfluidic device comprises:

Multiple 3rd input lines, in the plurality of 3rd input line, each is with the plurality of first One of input line is in fluid communication；

Multiple 4th input lines, in the plurality of 4th input line, each is with the plurality of second One of input line is in fluid communication；

The first control line with described first pressure source fluid communication；

Check valve, it is operable to block at least some of of described first control line；With

The second control line with described second pressure source fluid communication.

115, the microfluid system of item 114, wherein said multiple first input ports are operable to Receive UP source.

116, the microfluid system of item 114, wherein said multiple second input ports are operable to Receive UP source.

117, the microfluid system of item 114, it also comprises and is operable to block described second control At least one of second check valve of line processed.

118, the microfluid system of item 114, wherein said microfluidic device also comprises manifold, and it will The plurality of second input line in described carrier is multiple 4th defeated with described in described microfluidic device Enter line to be connected.

119, the microfluid system of item 118, its also comprise be operable to block described manifold it The valve group of part.

120, the microfluid system of item 114, wherein said microfluidic device also comprises multiple reative cell Group, one of each reative cell of each reative cell group and the plurality of 3rd input line and the plurality of the One of four input lines are in fluid communication.

Claims

1. a microfluidic device, it comprises:

First valve；

Second valve；

2. a microfluidic device, it comprises:

Check valve, it is arranged in described first control line.

3. a microfluidic device, it comprises:

Multiple reative cells；

4. the method operating the microfluidic device with multiple valve and check valve, described method includes:

5. the method operating the microfluidic device with multiple input port, described method includes:

Input fluid is provided to one of the plurality of input port；

Close the second valve group to separate multiple reative cells.

6. the method operating programmable microfluidic device, described microfluidic device has and the first input Line group and the reaction site array of the second input line group fluid communication, described method includes:

Start the 3rd valve group being operable to separate described reaction site.

7. a microfluidic device, it comprises:

8. the method operating programmable microfluidic device, described microfluidic device has reaction site battle array Row, itself and the first input line group, the second input line group and the manifold stream being connected described second input line group Body connects, and described method includes:

Start the 3rd valve group being operable to make described manifold disable；

Cancel and start described second valve group；

Start the 4th valve group being operable to separate described reaction site.

9. the method operating programmable microfluidic device, described programmable microfluidic device has and the One input line group and the reaction site array of the second input line group fluid communication, described method includes:

Cancel and start described second valve group；

Start described second valve group.

10. a microfluidic device, it comprises:

Multiple reaction site；

11. 1 kinds of microfluid systems, it comprises:

Carrier, it comprises:

Multiple first input ports；

Multiple second input ports；

First pressure source；With

Second pressure source