CN1696694A - Transmission through applied microflow liquid, and method of analyzing system - Google Patents
Transmission through applied microflow liquid, and method of analyzing system Download PDFInfo
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- CN1696694A CN1696694A CN 200510068206 CN200510068206A CN1696694A CN 1696694 A CN1696694 A CN 1696694A CN 200510068206 CN200510068206 CN 200510068206 CN 200510068206 A CN200510068206 A CN 200510068206A CN 1696694 A CN1696694 A CN 1696694A
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Abstract
A microfluid conveying and analyzing system comprises a flowing control unit with capillary channels, plastic fluid device of external connection type with linear actuation and reaction chamber with filling device. The system can be used in immunization analysis and DNA hybridization analysis or can be used to synthetize a series of digonucleotides in reaction chamber.
Description
The application is for dividing an application, and the applying date of original bill application is: on May 1st 2,2004, application number is: 200410043573.9, and invention and created name is: microfluid transmits and analytic system.
Technical field
The present invention relates to a kind of fluid and transmit and analyze a casket and a circumscribed linear actuators.Particularly a kind of system that carries out distinct program in microfluid transmission and analysis casket, this program comprises screening, immunology diagnosis, DNA diagnosis.
Background technology
In recent years, the highly-parallel method has been used for the analysis of biological substance (for example protein and DNA) by development.A large amount of different bound substances can be immobilized in solid surface, and can the highly-parallel mode record at the cross reaction of these materials and other compound.In recent years, although the size of solid surface is considerably reduced, and the density that is immobilized species is also increased dramatically, but typically, this kind analysis still needs some fluid operated steps, under not having fluid operated automaton or similarly installing, make its robotization become difficult.
Recently, developed some microfluidic platforms, solving these fluid operated problems, reduced reaction reagent consumption and increase the speed of these methods.The example of this kind platform is described in United States Patent (USP) the 5th, 856, and 174 and 5,922, in No. 591.As the Tranducers ' 97 of people such as Anderson in 1997, " microfluid biochemical analysis system " 477-480 page or leaf of being delivered in the agenda of 1997 International Conference on Solid-State Sensors and Actuators is described, and this device showed afterwards can carry out nucleic acid extraction, amplification and hybridization at the HIV viral sample.By the source pressure of valve, hydrophobicity vent port and the difference of utilizing gas control, liquid reactants can be operated to carry out foranalysis of nucleic acids in the microfluid casket.
Another example of this kind microfluidic platforms is to be described in United States Patent (USP) the 6th, 063, and in No. 589, wherein the use of centripetal force system is used for the capillary network of perfusion fluid to be included in liquid fluid casket on the CD by one.Passive breach valve is to flow carrying out bioanalysis in order to controlling liquid, " the Centrifugal Microfluidics:Applicaions " that is delivered as the Micro Total Analysis System2000 of people such as Kellog in uTas 2000 discussion agendas in 2000 narration.The further utilization of passive surface and microfluidic devices miniature at this kind has been described in United States Patent (USP) the 6th, 296, in No. 020, with the liquid of control micro device.
Another kind of pressure-driven liquid operating device is the utilization by electric field, moves with controlling liquid and molecule.Utilize these electrical means driving reactant, and utilize electrophoresis method in this kind system, to separate and carry out in the particular analysis by liquid medium, microfluid transmit and analytic system in have many work and be done.Utilize the device of this kind method to be described in United States Patent (USP) the 4th, 908, in No. the 5th, 858,804, No. 112, No. the 6th, 033,544, United States Patent (USP) and the United States Patent (USP).
Other utilizes static valve matrix (United States Patent (USP) the 6th, 240, No. 944), iron content fluid Micropump (United States Patent (USP) the 6th, 318, No. 970), and the microfluidic procedures device of fluid mobility regulator (United States Patent (USP) the 5th, 839, No. 467) is also narrated.
The application of this kind microfluidic procedures device, having increases the potentiality of analyzing productive rate, reduction reactant consumption, simplifying diagnostic device and reduction analysis cost.
Summary of the invention
The object of the present invention is to provide a kind of microfluid that carries out analysis of fluid sample immunology diagnosis or DNA analysis program to transmit and analytic system.
System of the present invention comprises that one has the plastic liquid device that at least one sees through capillary channel and circumscribed linear actuators, and it is connected to the reaction chamber of reperfusion structure.This device comprises two plastic basic units, one first basic unit and one second basic unit, and it contains capillary channel, reaction chamber and pump/valve chamber, and a flexible central dividing plate between first and second basic unit, it can provide the airtight interface of fluidic structures and valve and pump dividing plate.The passive check valve structure forms in this three bed device, and it is by a kind of gas or flow element of being used for being provided, utilizing the bending of central dividing plate, and the passage of going to a grassroots level certainly flow to the passage of first basic unit.Moreover in the opposite direction mobile is with second basic unit restriction dividing plate flexure operation.Perhaps, the passive check valve structure can be made into by touching this apparatus structure and can make first basic unit flow to second basic unit.Pump configuration is to be formed by combination pump chamber and two passive check valve structures of operating equidirectionals in the formation of this device.At bottom hole corresponding to pump chamber is arranged also.Therefore linear actuators-in the outside of plastic liquid device-can place this hole with crooked pump central dividing plate can provide the fluid of pumping action to the device.This kind pump configuration is original to be unidirectional just.
Beneficial effect of the present invention is: in a preferred embodiments, said system can be used for carrying out immunoassay, it is with the pump perfusion different reactant from the inlet storagetank, contains one or more immobilized antibody or antigen-reactive chamber by one, flow to outlet at last.In another preferred embodiments, this system can be used for carrying out the detection of DNA analysis, for example to the hybridization of the dna probe that is fixed on reaction chamber.This system is used in synthetic a series of oligonucleotides in the reaction chamber.Carry out solid phase reaction in the reaction chamber and provide dispersion to be dispersed to reaction chamber from the differential responses thing of reaction chamber and with the differential responses thing although system of the present invention is highly suitable for, it is not to be limited to only to carry out solid phase reaction.
System of the present invention also is suitable for abandoning the formula diagnostic application very much.The utilization of native system can reduce consumables to only for the plastic liquid casket, and remove general when a large amount is used the fixing cross-contamination issue that causes of pipette tip formula robotization dropper of use.
Description of drawings
Figure 1A is the vertical view at the pump configuration of plastic liquid device of the present invention.
Figure 1B is the cross-sectional view at the pump configuration of plastic liquid device of the present invention.
Single fluid transmits Fig. 2 and the vertical view of analytical equipment for plastic liquid device of the present invention is assembled into.
Five kinds of fluids transmit Fig. 3 and the vertical view of analytical equipment for plastic liquid device of the present invention is assembled into.
Three kinds of fluids of recycle transmit Fig. 4 and the vertical view of analytical equipment for plastic liquid device of the present invention is assembled into.
Embodiment
Fluid of the present invention transmits and analytic system, comprise a fluid casket, it comprises first basic unit, second basic unit and the airtight flexible middle layer dividing plate that is positioned at the interface between this first and second basic unit with the passage that forms one or more capillary dimensions therein, at least one fluid storage tank, at least one interface, at least one pump chamber, and at least one reaction chamber that is connected to this pump chamber through these pipelines; One fluid FLOW CONTROL structure, it forms in this fluid casket, and limit fluid only uniaxially flows through this reaction chamber via these pipelines; And a linear brake, it ties up to a pumping action is provided in this pump chamber, so that this fluid is poured into to this interface by this reaction chamber and these pipelines from this fluid storage tank.
The present invention provides a kind of fluid means that is used for fluid transmission and analytic system in addition, it comprises: first basic unit, second basic unit and the airtight flexible middle layer dividing plate that is positioned at the interface between this first and second basic unit reach at least one reaction chamber that is connected to these pipelines to form the passage of one or more capillary dimensions therein; And limit fluid only uniaxially flow through the element of this reaction chamber via these pipelines.
Figure 1B is presented at the cross-sectional view of the pump configuration of plastic liquid device of the present invention.Shown in Figure 1B, this plastic liquid casket comprises three main basic units: first basic unit 21, second basic unit 22, and a flexible middle layer 23.Can be assembled for example screw assembling, hot pressing, ultrasonic combination, clamping or suitable reactivity/stickability associated methods for these three layers by different plastics assemble methods.This first and second basic unit contains all that different definition is the feature of capillary dimensions passage and pump chamber, valve chamber, reaction chamber, storagetank and inlet/outlet in casket.Figure 1A is the vertical view of the pump configuration of Figure 1B.This pump configuration is defined as a pump chamber 14 and two passive check valves 15 and can provides high impedance to make to flow only for unidirectional.Passive check valve 15 comprises one second basic unit's passage 13 and one first basic unit's passage 11, is separated by the middle layer, makes hole by middle layer 12 be included in first basic unit's passage 11 but not in second basic unit's passage 13.This kind passive check valve structure can provide Low ESR to make gas/fluid flow to first basic unit's passage 11 from second basic unit's passage 13, similarly also provides high impedance to make gas/fluid flow to second basic unit's passage 13 from first basic unit's passage 11.Pump chamber 14 have one first basic unit chamber and one at the inlet hole of second basic unit to discharge the middle layer as dividing plate.The linear actuators 24 that is connected on the fluid casket outward can provide necessary strength to make the dividing plate distortion.Wherein each of these two passive test valves comprises one first basic unit's pipeline and one second basic unit's pipeline of being separated by this middle layer, wherein is included in this first basic unit pipeline by the hole system that forms in this middle layer but is not included in the second basic room pipeline.
Fig. 2 shows that plastic cement fluid means of the present invention is assembled into the vertical view of single fluid transmission and analytical equipment.At first utilize dropper or similar equipment to put into storagetank 31 with artificial or automated manner fluid, the pump configuration 32 of similar Figure 1A is included in this device.By repeatedly activating outside linear actuators, the fluid in storagetank 31 is through pump perfusion process pump configuration 32, capillary channel 33 and enter reaction chamber 34.Reaction chamber 34 comprises that several and this fluid carries out the immobilized biomolecule 35 of unique solid phase reaction.After the specific reaction time, this fluid passes through reaction chamber 34 and pass-out opening 36 through the pump perfusion.
First and second basic unit of plastic cement fluid casket of the present invention can utilize different plastic materials, and for example methyl methacrylate, polystyrene, polycarbonate, polypropylene or tygon chlorine are made.Because the optical characteristics of reaction result in reaction chamber, first basic unit is preferably with the clear plastic material made.Kapillary, reaction chamber and pump chamber for example can utilize, and the method for injection moulding, compression molding, hot forming or machining forms in this kind basic unit.The thickness of first and second basic unit is fit to but is not limited in 1 millimeter thickness to 3 millimeters scopes.Flexible middle layer can multiple polymers and elastomeric material form, for example latex, silicon elastic body, tygon chlorine and fluoroelastomer form.Form the method for this kind feature in the middle layer, comprise cross cutting, rotating die cutting, laser carved, injection moulding and reaction injection molding.
Linear actuators of the present invention is preferably but is not restricted to o.Other linear actuators that is fit to comprises motor/cam/piston structure, the linear activated gas of piezoelectricity and motor/linear gear structure.
The present invention provides a kind of method of fluid sample immunoassay that system of the present invention contains several biomolecule of unknown concentration of using in addition, and it comprises the following steps:
(a) with this fluid sample or should put at least one fluid storage tank to the fluid casket with the fluid sample of conjugation polymer blends;
(b) with this fluid sample or should with the fluid sample of conjugation polymer blends from this fluid storage tank contain to this fluid casket a kind of with the pump perfusion or several the immobilized antigen or the reaction chamber of antibody in;
(c) make this fluid sample or should with fluid sample and this immobilized antigen or one section predetermined reaction time of antibody response of conjugation polymer blends; And
(d) this fluid sample or this were discharged by escape hole from this reaction chamber with the fluid sample of conjugation polymer blends.
Preferably, this method can further comprise the following steps:
(e) will contain with the specific secondary antibody solution that can detect molecular conjugationization and put to this fluid storage tank;
(f) this antibody-solutions is poured into to this reaction chamber with pump from this fluid storage tank;
(g) after one section predetermined reaction time, this antibody-solutions is discharged by escape hole with the pump perfusion;
(h) produce the signal that to detect.
In addition, this method can comprise further that at least one cleaning buffer solution that will be placed in this fluid storage tank reaches the cleaning step of discharging by escape hole with the pump perfusion by this reaction chamber.Or in step (d) afterwards, further comprise the matrix damping fluid is put into the step that makes that matrix and this mix with the fluid sample of conjugation fusion.
In said method, this fluid storage tank, this reaction chamber and this escape hole are that the passage by one or more capillary dimensions is connected, wherein this fluid casket comprises first basic unit, second basic unit and airtightly is positioned at interface between this first and second basic unit to form the flexible middle layer dividing plate of this fluid storage tank, these pipelines, this reaction chamber and this escape hole therein, wherein this fluid casket further provide fluid FLOW CONTROL structure with limit this fluid sample only uniaxially flow through this reaction chamber via these pipelines.
The present invention provides a kind of in addition and uses system of the present invention and contain one or the sample immune analysis method of the biomolecule of several unknown concentration, and it comprises the following steps:
(a ') puts at least one sample storagetank to the fluid casket with this sample;
(b ') puts damping fluid storagetank to the fluid casket with cleaning buffer solution;
(c ') in this fluid casket, provide an air purge storagetank that keeps sky so that air purge to be provided;
(d ') will put to a matrix storagetank of this fluid casket the special matrix solution of secondary antibody conjugate;
(e ') the secondary antibody conjugate is put to an antibody storagetank of this fluid casket;
(f ') this sample, this cleaning buffer solution, this matrix solution and this secondary antibody conjugate are passed through one section predetermined reaction time of first reaction chamber with the pump configuration perfusion;
(g ') provide one second reaction chamber to go out in contrast and by non-return valve this sample separation from this first storagetank;
(h ') confirm the immunoassay result by measuring method.
Preferably, step in the method (f ') further can comprise the following steps:
(f-1 ') this sample is full of this first reaction chamber with pump perfusion to this first reaction chamber up to this sample from this sample storagetank;
Pour into this sample to this waste liquid tank with pump from this first storagetank after one section first predetermined reaction time (f '-2);
Pour into this cleaning buffer solution to this first reaction chamber with pump from this damping fluid storagetank (f '-3);
Pour into this secondary antibody conjugate to this first reaction chamber with pump from this antibody storagetank (f '-4);
Is pouring into this secondary antibody conjugate to this waste liquid tank with pump from this first reaction chamber after one section second predetermined reaction time (f '-5);
Pour into this matrix solution to this first reaction chamber with pump from this matrix storagetank (f '-6); And
Is pouring into this matrix solution to this waste liquid tank with pump from this first reaction chamber after one section the 3rd predetermined reaction time (f '-7), and this first reactive tank was replaced should ask the cleaning buffer solution of wash buffer storagetank certainly.
In said method, wherein this fluid storage tank and this reaction chamber are connected by the passage of one or more capillary dimensions, wherein this fluid casket comprises first basic unit, second basic unit and airtightly be positioned at interface between this first and second basic unit to form this fluid storage tank therein, this damping fluid storagetank, this air purge storagetank, this matrix storagetank, this antibody storagetank, the flexible middle layer dividing plate of these pipelines and this reaction chamber, wherein this fluid casket further provides fluid FLOW CONTROL structure to limit this sample, this cleaning buffer solution, this matrix solution, this secondary antibody conjugate only uniaxially flows through this reaction chamber via these pipelines.
The present invention provides a kind of method that system of the present invention carries out the DNA hybridization analysis of using in addition, and it comprises the following steps:
(a ") is fixed on reaction chamber in the fluid casket with several dna probes;
(b ") will contain one or more fluid sample through the unknown nucleotide sequence DNA of fluorescent labelling and amplification to be put to the sample storagetank of fluid casket;
(c ") is put the first stringent wash buffer liquid to the first cleaning buffer solution storagetank of this fluid casket;
(d ") is put the second stringent wash buffer liquid to the second cleaning buffer solution storagetank of this fluid casket;
(e ") keeps a reaction chamber under a steady temperature;
(f ") with this fluid sample from this sample storagetank with pump perfusion by this reaction chamber one section predetermined hybridization time to the circulation storagetank of this fluid casket;
(g ") discharges from this circulation storagetank and this reaction chamber this fluid sample with the pump perfusion;
(h ") one section first is scheduled to scavenging period with pump perfusion to this circulation storagetank and this first stringent wash buffer liquid that circulates by this reaction chamber from this first cleaning buffer solution storagetank with this first stringent wash buffer liquid;
(i ") discharges from this circulation storagetank and this reaction chamber this first stringent wash buffer liquid with the pump perfusion;
(j ") one section second is scheduled to scavenging period with pump perfusion to this circulation storagetank and this second stringent wash buffer liquid that circulates by this reaction chamber from this second cleaning buffer solution storagetank with this second stringent wash buffer liquid;
(k ") discharges from this circulation storagetank and this reaction chamber this second stringent wash buffer liquid with the pump perfusion; And
(l ") reaches DNA hybridization.
In said method, this sample storagetank wherein, this first buffer solution for cleaning storagetank, the second buffer solution for cleaning storagetank, this circulation storagetank and this reaction chamber are that the passage by one or more capillary dimensions is connected, wherein this fluid casket comprises first basic unit, second basic unit and airtightly be positioned at interface between this first and second basic unit to form this sample storagetank therein, this first cleaning buffer solution storagetank, the second cleaning buffer solution storagetank, the flexible middle layer dividing plate of this circulation storagetank and this reaction chamber, wherein this fluid casket further provides fluid FLOW CONTROL structure to limit this fluid sample, this first and second stringent wash buffer liquid only uniaxially flows through this reaction chamber via these pipelines.
The present invention will utilize plastic cement fluid casket of the present invention and external actuator, further describe in the example of the different different structures of analyzing is carried out in a series of narrations.
The present invention is further described in detail via the following example, but these embodiment are not in order to the scope of the invention is imposed any restrictions only in order to illustrate concrete enforcement aspect of the present invention.Modification that any expert who is familiar with this technology can reach easily and change all are included in the scope of instructions and claim.
Embodiment 1: immunoassay
The plastic cement fluid casket of Fig. 2 can be in reaction chamber 34 one or more antibody 35 of immobilization and be used to carry out immunoassay.At first, will contain one or more antigen of unknown concentration or antibody or biomolecule nucleic acid and put into storagetank 31, and then linear actuators repeatedly be driven sample is poured into to reaction chamber 34 with pump from storagetank 31.Make sample and one section setting-up time of immobilized antibody 35 reactions then.After the reaction time of one section setting, then sample autoreaction chamber 34 is discharged through outlet 36.Then cleaning buffer solution is put into storagetank 31 and repeated to drive outside linear actuators cleaning buffer solution is exported 36 with the pump perfusion through reaction chamber 34 and discharge.This kind cleaning step can optionally repeat.Contain and can detect molecule, the specific secondary antibody solution of horseradish peroxidase ferment, alkaline phosphatase ferment and fluorescent label conjugation for example, put into storagetank 31, then secondary antibody solution is repeatedly driven linear actuators and pour into to reaction chamber 34 with pump.After one period schedule time, solution is discharged outlet 36 with the pump perfusion.Then reaction chamber 34 is cleaned with aforesaid similar manner.When using the ferment conjugate, matrix solution is put into storagetank 31 and pour into to reaction chamber 34 with pump.Matrix will be reacted with the ferment that provides the immobilized antibody reaction that can detect molecule to be caught with previous then.For promoting analytical effect, reaction chamber 34 can maintain under constant 37 ℃.
According to the present invention, plastic cement fluid casket does not need structure to dress up single fluid transmission and analytical equipment.Fig. 3 shows that structure dresses up that five kinds of fluids transmit and the plastic cement casket of analytical equipment.This kind device can carry out immunoassay by provide immobilized antibody in reaction chamber 34.This reaction chamber not structure dress up the wide-angle rectangular area, and be the bending channel space of similar capillary dimensions.This kind configuration provides the more uniform reaction chamber that flows through with the space of waste.For carrying out immunoassay, one or more antibody that will contain unknown concentration is put into storagetank 34, and cleaning buffer solution is put into storagetank 42.It is empty so that air purge to be provided that storagetank 43 keeps, and will put into storagetank 45 to the special matrix solution of secondary antibody conjugate.All storagetanks are connected to the storagetank that is also connected certainly similar in appearance to the pump configuration of Fig. 1 provides pump to pour into to waste liquid tank 49 through reaction chamber 46.Other provides second reaction chamber 47 in contrast, and is gone out by the sample separation of non-return valve 48 with storagetank.The method of carrying out immunoassay in this device is equivalent to aforementionedly be used for the single fluid configuration, but different be that the reaction reagent system of every kind of individual separation is included in other storagetank, and utilize other outer lines actuator with individual other pump configuration perfusion.At first, be connected to storagetank 41 and repeatedly drive, fill up reaction chamber 46 up to sample with respect to the outside linear actuators of pump.After one section predetermined reaction time, the sample utilization is connected to respect to the pump of sample storagetank 41 or the pump that is connected to air purge groove 43 pours into to waste liquid storagetank 49.Subsequently with cleaning buffer solution by repeatedly driving the outside linear actuators be connected to respect to the pump configuration that cleans storagetank 42, pour into to reaction chamber 46 with pump.Clean cycle and air purge can optionally repeat.Then with secondary antibody by repeatedly driving the outside linear actuators be connected to respect to the pump configuration of storagetank 45, pour into to reaction chamber 46 with pump.After one section predetermined reaction time, discharge the pump autoreaction chamber 46 that this secondary antibody utilization is connected to the pump of storagetank 45 or is connected to air purge groove 43.Reaction chamber 46 is subsequently to clean as previous mode.Matrix by repeatedly driving the outside linear actuators that is connected to respect to the pump configuration of storagetank 44, is poured into to reaction chamber 46 with pump.After one section predetermined reaction time, matrix autoreaction chamber discharged and replace with the cleaning buffer solution of storagetank 42.The result of immunoassay can see through first basic unit subsequently and determine with measuring method.
Moreover, with the reaction that plastic cement fluid casket of the present invention carries out, the reaction that need not be defined as in inactive liquid to be carried out.Fig. 4 shows according to the present invention that the continuous fluid that provides through reaction chamber is provided plastic cement fluid casket and moves.In this configuration, storagetank 51,52 and 53 is connected to similar in appearance to the individual pump structure of five kinds of fluid configurations of Fig. 3, but is circulation storagetank 56 in the middle of being connected at this.Pump configuration 57 is connected to circulation storagetank 56 and circulates continuously so that fluid to be provided, and self-loopa storagetank 56 flows back in the circulation storagetank 56 by reaction chamber 55 again.In this way, fluid is capable of circulation does not stop by reaction chamber.This kind fluid moves can provide mixed effect preferably, faster the reaction time and make sample and the fixed compound qualitative response of reaction chamber 55 complete.Make it can provide fluid self-loopa storagetank 56 to pour into to waste liquid tank 54 pump configuration 58 connections with pump.Can in this device, carry out by sessile antibody in reaction chamber 55 similar in appearance to aforesaid immunoassay.
The antigen or the antibody sample that will contain unknown concentration are put into circulation storagetank 56, and secondary antibody conjugate solution is put into storagetank 52, matrix solution is put into storagetank 53, and cleaning buffer solution is put into storagetank 51.Remaining method is identical with preceding method, and adds that diverting the fluid to circulation storagetank 56 and self-loopa storagetank 56 shifts, and circulation continuously in all reaction time.
Example 2:DNA hybridization
System of the present invention also can be used for carrying out the DNA hybridization analysis.Utilize the plastic cement fluid casket of Fig. 4, one or more dna probe is fixed in the reaction chamber 55.The DNA fluid sample that will contain the unknown nucleotide sequence of one or more fluorescent labelling and amplification is put into storagetank 52.The first strict cleaning solution is put into storagetank 51, the second strict cleaning solution is put into storagetank 53.Reaction chamber maintained under 52 ℃ the steady temperature.Sample by repeatedly driving the linear actuators that is connected to respect to the pump configuration of storagetank 52, is passed in the circulation storagetank 56.Then with sample by repeatedly driving the linear actuators be connected to respect to pump configuration 57, circulation is through reaction chamber 55.With one section predetermined hybridization time of the continuous circulation of sample, be generally 30 minutes to two hours.Then sample is utilized driving pump structure 57 and 58 to discharge with opposite way self-loopa storagetank 56 and reaction chamber 55.Then with the first strict cleaning solution by the linear actuators that repeatedly drives corresponding to the pump configuration that is connected to storagetank 51, be passed in the circulation storagetank.Subsequently damping fluid is circulated through reaction chamber 55 with aforementioned same way as.Behind one section predetermined scavenging period, damping fluid autoreaction chamber 55 and circulation storagetank 56 are discharged.Then the second stringent wash buffer liquid is also circulated through reaction chambers 55 to be passed to circulation storagetank 56 similar in appearance to aforesaid mode.After second cleaning buffer solution was discharged, the result of DNA hybridization can the fluorescent development, colourimetry, luminescence detection or biotin-streptavidin-ferment detect and read.
Claims (29)
1. an applicating fluid transmits and analytic system contains the method for fluid sample immunoassay of several biomolecule of unknown concentration, it is characterized in that, comprises the following steps:
(a) with this fluid sample or should put at least one fluid storage tank to the fluid casket with the fluid sample of conjugation polymer blends;
(b) with this fluid sample or should contain to this fluid casket with the pump perfusion in several the immobilized antigen or the reaction chamber of antibody from this fluid storage tank with the fluid sample of conjugation polymer blends;
(c) make this fluid sample or should with fluid sample and this immobilized antigen or one section predetermined reaction time of antibody response of conjugation polymer blends; And
(d) with this fluid sample or should discharge from this reaction chamber with the fluid sample of conjugation polymer blends and pass through escape hole.
2. method as claimed in claim 1 is characterized in that, further comprises the following steps:
(e) will contain with the specific secondary antibody solution that can detect molecular conjugationization and put to this fluid storage tank;
(f) this antibody-solutions is poured into to this reaction chamber with pump from this fluid storage tank;
(g) after one section predetermined reaction time, this antibody-solutions is discharged this opening with the pump perfusion;
(h) provide the signal that to detect.
3. method as claimed in claim 1, it is characterized in that further comprising be placed on cleaning buffer solution in this fluid storage tank with the pump perfusion by this reaction chamber and discharge the cleaning step of this opening.
4. method as claimed in claim 1 is characterized in that, (d ") further comprises the matrix damping fluid is put into the step that makes that matrix and this mix with the fluid sample of conjugation polymer blends afterwards in step.
5. method as claimed in claim 2 is characterized in that, in step (d) and step (g) afterwards, further comprise be placed on cleaning buffer solution in this fluid storage tank with the pump perfusion by this reaction chamber and discharge the cleaning step of this opening.
6. as the method for claim 1 or 3, it is characterized in that wherein this fluid-like contains multiple different antibody.
7. as the method for claim 2 or 4, it is characterized in that, wherein this can to detect molecule be to be selected to comprise horseradish peroxidase ferment, alkaline phosphatase ferment and fluorescent group that label constitutes.
8. as the method for claim 2 or 4, it is characterized in that wherein this antibody-solutions is a matrix solution, can with in step (g) but in be provided any ferment reaction that the immobilized antibody of detection signal is caught.
9. as the method for claim 1 or 3, it is characterized in that, wherein this fluid storage tank, this reaction chamber and this escape hole are that pipeline by one or more capillary dimensions is connected, wherein this fluid casket comprises first basic unit, second basic unit and airtightly is positioned at interface between this first and second basic unit to form the flexible middle layer dividing plate of this fluid storage tank, these pipelines, this reaction chamber and this escape hole therein, wherein this fluid casket further provide fluid FLOW CONTROL structure with limit this fluid sample only uniaxially flow through this reaction chamber via these pipelines.
10. as the method for claim 1 or 3, it is characterized in that, wherein in this pump perfusion step (b) and (d), this fluid-like strain provides pumping action by the pump chamber of linear actuators in the fluid casket, and this fluid is passed through this reaction chamber and arrive this escape hole with the pump perfusion via these pipelines from this fluid storage tank.
11. method as claim 2 or 4, it is characterized in that, this fluid storage tank wherein, this reaction chamber and this escape hole system are connected by the pipeline of one or more capillary dimensions, wherein this fluid casket comprises first basic unit, second basic unit and airtightly be positioned at interface between this first and second basic unit to form this fluid storage tank therein, these pipelines, the flexible middle layer dividing plate of this reaction chamber and this escape hole, wherein this fluid casket further provide fluid FLOW CONTROL structure with limit this fluid sample and antibody-solutions only uniaxially flow through this reaction chamber via these pipelines.
12. method as claim 10, it is characterized in that, wherein in this pump perfusion step (b), (d), (f) reach (g), this fluid sample and this antibody-solutions system provide pumping action by the pump chamber of linear actuators in the fluid casket respectively, and this fluid and this antibody-solutions are passed through this reaction chamber to this escape hole with the pump perfusion via these pipelines from this fluid storage tank.
13. method as claimed in claim 9, it is characterized in that, wherein this pump chamber contains a matrix chamber that forms and a hole that forms in second basic unit in this first basic unit, to discharge this middle layer with as pump middle layer dividing plate, wherein this linear brake moves with crooked this pump middle layer dividing plate in this hole and therefore provides a kind of necessary strength so that this pump middle layer dividing plate distortion, with in this pump chamber, provide this pumping action with this fluid sample from this fluid storage tank with the pump perfusion via these pipelines by this reaction chamber to this escape hole.
14. method as claim 11, it is characterized in that, wherein this pump chamber contains a matrix chamber that forms and a hole that forms in second basic unit in this first basic unit, to discharge this middle layer with as pump middle layer dividing plate, wherein this linear brake moves with crooked this pump middle layer dividing plate in this hole and therefore provides a kind of necessary strength so that this pump middle layer dividing plate distortion, with in this pump chamber, provide this pumping action with this fluid sample from this fluid storage tank with the pump perfusion via these pipelines by this reaction chamber to this escape hole.
15. method as claimed in claim 9, it is characterized in that, wherein this fluid FLOW CONTROL structure comprises that two are positioned at this pump chamber passive test valve before and after in this fluid casket, flow to this escape hole from this fluid storage tank by this reaction chamber and via these pipelines to provide, and provide higher resistance to this fluid sample to flow to this fluid storage tank from this escape hole than Low ESR to this fluid sample.
16. method as claim 11, it is characterized in that, wherein this fluid FLOW CONTROL structure comprises that two are positioned at this pump chamber passive test valve before and after in this fluid casket, flow to this escape hole from this fluid storage tank by this reaction chamber and via these pipelines to provide, and provide higher resistance to this fluid sample and this antibody-solutions to flow to this fluid storage tank from this escape hole than Low ESR to this fluid sample and this antibody-solutions.
17. method as claimed in claim 9, it is characterized in that, wherein this fluid FLOW CONTROL structure comprises two passive test valves in this fluid casket, with the limit fluid sample from another this passage to this first basic unit of this channel flow in this second basic unit, it is that bending by this pump middle layer dividing plate flows to this escape hole to control this fluid sample from this fluid storage tank, and wherein any this fluid storage tank system that flows back into from this fluid sample of this escape hole is controlled by limit the crooked of this pump middle layer dividing plate with this second basic unit.
18. method as claim 11, it is characterized in that, wherein this fluid FLOW CONTROL structure comprises two passive test valves in this fluid casket, with limit fluid sample and this antibody-solutions from another this passage to this first basic unit of this channel flow in this second basic unit, its be by the bending of this pump middle layer dividing plate to control this fluid sample and this antibody-solutions flows to this escape hole from this fluid storage tank, wherein controlled by limit the crooked of this pump middle layer dividing plate with this second basic unit from this fluid sample of this escape hole and any this fluid storage tank that flows back into of this antibody-solutions.
19. an applicating fluid transmits and analytic system contains the method for sample immunoassay of several biomolecule of unknown concentration, it is characterized in that, comprises the following steps:
(a ') puts at least one sample storagetank to the fluid casket with this sample;
(b ') puts damping fluid storagetank to the fluid casket with cleaning buffer solution;
(c ') in this fluid casket, provide an air purge storagetank that keeps sky so that air purge to be provided;
(d ') will put to a matrix storagetank of this fluid casket the special matrix solution of secondary antibody conjugate;
(e ') the secondary antibody conjugate is put to an antibody storagetank of this fluid casket;
(f ') this sample, this cleaning buffer solution, this matrix solution and this secondary antibody conjugate are passed through one section predetermined reaction time of first reaction chamber with the pump configuration perfusion;
(g ') provide one second reaction chamber to go out in contrast and by test valve this sample separation from this first storagetank;
(h ') confirm the immunoassay result by optical measurement.
20. the method as claim 18 is characterized in that, wherein this step (f ') further comprises the following steps:
(f '-1) is full of this first reaction chamber with pump perfusion to this first reaction chamber up to this sample from this sample storagetank with this sample;
Pour into this sample to this waste liquid tank with pump from this first storagetank after one section first predetermined reaction time (f '-2);
Pour into this cleaning buffer solution to this first reaction chamber with pump from this damping fluid storagetank (f '-3);
Pour into this secondary antibody conjugate to this first reaction chamber with pump from this antibody storagetank (f '-4);
Is pouring into this secondary antibody conjugate to this waste liquid tank with pump from this first reaction chamber after one section second predetermined reaction time (f '-5);
Pour into this matrix solution to this first reaction chamber with pump from this matrix storagetank (f '-6); And
(f '-7) with this matrix solution after one section the 3rd predetermined reaction time from this first reaction chamber with pump perfusion to this waste liquid tank, and this first reactive tank replaced with the cleaning buffer solution of this cleaning buffer solution storagetank certainly.
21. method as claim 19, it is characterized in that, wherein this pump configuration comprises that several are connected to the pump of this sample storagetank, this cleaning buffer solution groove, this air purge storagetank, this matrix storagetank and this antibody storagetank respectively, respectively this sample, this cleaning buffer solution, this matrix solution and this secondary antibody conjugate is passed through this first reactive tank to this waste liquid storagetank with the pump perfusion.
22. method as claim 19, it is characterized in that, wherein the sample storagetank is a circulation storagetank, and this pump configuration is connected to this circulation storagetank and is circulated in this waste liquid storagetank continuously by this first reaction chamber and flows back in this circulation storagetank from this circulation storagetank so that liquid to be provided, and makes this sample, this cleaning buffer solution, this matrix solution and this secondary antibody conjugate can cycle through this first reaction and does not stop.
23. as claim 19,20 or 21 method, it is characterized in that, wherein this fluid storage tank and this reaction chamber are connected by the pipeline of one or more capillary dimensions, wherein this fluid casket comprises first basic unit, second basic unit and airtightly be positioned at interface between this first and second basic unit to form this fluid storage tank therein, this damping fluid storagetank, this air purge storagetank, this matrix storagetank, this antibody storagetank, the flexible middle layer dividing plate of these pipelines and this reaction chamber, wherein this fluid casket further provides fluid FLOW CONTROL structure to limit this sample, this cleaning buffer solution, this matrix solution, this secondary antibody conjugate only uniaxially flows through this reaction chamber via these pipelines.
24. method as claim 22, it is characterized in that, wherein in this pump perfusion step (f '), this sample, this cleaning buffer solution, this matrix solution and this secondary antibody conjugate, provide pumping action by the pump chamber of several linear actuators in the fluid casket respectively, this sample, this cleaning buffer solution, this matrix solution and this secondary antibody conjugate are passed through this reaction chamber with the pump perfusion via these pipelines from this sample storagetank, this damping fluid storagetank, this air purge storagetank, this matrix storagetank and this antibody storagetank respectively.
25. an applicating fluid transmits and analytic system is carried out the method for DNA hybridization analysis, it is characterized in that, comprises the following steps:
(a ") is fixed on reaction chamber in the fluid casket with several dna probes;
(b ") will contain one or more fluid sample through the unknown nucleotide sequence of fluorescent labelling and amplification DNA to be put to the sample storagetank of fluid casket;
(c ") is put the first stringent wash buffer liquid to the first cleaning buffer solution storagetank of this fluid casket;
(d ") is put the second stringent wash buffer liquid to the second cleaning buffer solution storagetank of this fluid casket;
(e ") keeps a reaction chamber under a steady temperature;
(f ") with this fluid sample from this sample storagetank with pump perfusion by this reaction chamber one section predetermined hybridization time to the circulation storagetank of this fluid casket;
(g ") discharges from this circulation storagetank and this reaction chamber this fluid sample with the pump perfusion;
(h ") one section first is scheduled to scavenging period with pump perfusion to this circulation storagetank and this first stringent wash buffer liquid that circulates by this reaction chamber from this first cleaning buffer solution storagetank with this first stringent wash buffer liquid;
(i ") discharges from this circulation storagetank and this reaction chamber this first stringent wash buffer liquid with the pump perfusion;
(j ") one section second is scheduled to scavenging period with pump perfusion to this circulation storagetank and this second stringent wash buffer liquid that circulates by this reaction chamber from this second cleaning buffer solution storagetank with this second stringent wash buffer liquid;
(k ") discharges from this circulation storagetank and this reaction chamber this second stringent wash buffer liquid with the pump perfusion; And
(l ") reaches DNA hybridization.
26. method as claim 24, it is characterized in that, this sample storagetank wherein, this first buffer solution for cleaning storagetank, the second buffer solution for cleaning storagetank, this circulation storagetank and this reaction chamber are that the pipeline by one or more capillary dimensions is connected, wherein this fluid casket comprises first basic unit, second basic unit and airtightly be positioned at interface between this first and second basic unit to form this sample storagetank therein, this first cleaning buffer solution storagetank, the second cleaning buffer solution storagetank, the flexible middle layer dividing plate of this circulation storagetank and this reaction chamber, wherein this fluid casket further provides fluid FLOW CONTROL structure to limit this fluid sample, this first and second stringent wash buffer liquid only uniaxially flows through this reaction chamber via these pipelines.
27. method as claim 25, it is characterized in that, wherein in this pump perfusion step (f ") in (k "), this fluid sample and this first and second stringent wash buffer liquid, provide pumping action by linear actuators at least one pump chamber in the fluid casket, this fluid sample and this first and second stringent wash buffer liquid are passed through this circulation storagetank and this reaction chamber with the pump perfusion via these pipelines from this sample storagetank, this first stringent wash buffer liquid storagetank, this second stringent wash buffer liquid storagetank.
28. method as claim 26, it is characterized in that, this this pump chamber hole of containing the matrix chamber that in this first basic unit, forms and in second basic unit, forming wherein wherein, to discharge this middle layer with as pump middle layer dividing plate, wherein this linear brake moves with crooked this pump middle layer dividing plate in this hole and therefore provides a kind of necessary strength so that this pump middle layer dividing plate distortion, with in this pump chamber, provide this pumping action with this fluid sample and first and second stringent wash buffer liquid from this sample storagetank, this first stringent wash buffer liquid storagetank, this second stringent wash buffer liquid storagetank passes through this circulation storagetank and this reaction chamber with the pump perfusion via these pipelines.
29. the method as claim 24 is characterized in that, wherein this is reached DNA hybridization system and carries out in fluorescent developments, look detection, luminous detection or biotin-streptavidin-ferment detection.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2005100682069A CN100504389C (en) | 2004-05-12 | 2004-05-12 | Transmission through applied microflow liquid, and method of analyzing system |
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| CNB2005100682069A CN100504389C (en) | 2004-05-12 | 2004-05-12 | Transmission through applied microflow liquid, and method of analyzing system |
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| CNB2004100435739A Division CN100394184C (en) | 2002-09-27 | 2004-05-12 | Micro-fluid transfer and analysis system |
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| CN1696694A true CN1696694A (en) | 2005-11-16 |
| CN100504389C CN100504389C (en) | 2009-06-24 |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102378595A (en) * | 2009-03-31 | 2012-03-14 | 森西勒Pat股份公司 | Medical device for measuring an analyte concentration |
| CN104220171A (en) * | 2012-02-21 | 2014-12-17 | 赛蒂奇道蒂奇公司 | Perfusion device, corresponding apparatus using said perfusion device and method to analyze the thrombotic-ischemic and hemorrhagic pathology |
| CN104568537A (en) * | 2014-11-05 | 2015-04-29 | 华文蔚 | Method for treating biological micro-fluidic sample |
| CN105628660A (en) * | 2015-12-29 | 2016-06-01 | 大连理工大学 | Passive microvalve POCT chip |
| CN108883414A (en) * | 2016-04-29 | 2018-11-23 | 建奥迪斯有限公司 | Method and component for molecule recycling |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SE502618C2 (en) * | 1994-10-03 | 1995-11-27 | Ericsson Telefon Ab L M | Method and apparatus for obtaining a wide dynamic range of a receiver in a digital radio transmission system |
| GB9502112D0 (en) * | 1995-02-03 | 1995-03-22 | British Biocell Int | Assay device and method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102378595A (en) * | 2009-03-31 | 2012-03-14 | 森西勒Pat股份公司 | Medical device for measuring an analyte concentration |
| CN104220171A (en) * | 2012-02-21 | 2014-12-17 | 赛蒂奇道蒂奇公司 | Perfusion device, corresponding apparatus using said perfusion device and method to analyze the thrombotic-ischemic and hemorrhagic pathology |
| CN104220171B (en) * | 2012-02-21 | 2017-01-18 | 赛蒂奇道蒂奇公司 | Perfusion device and corresponding apparatus using said perfusion device |
| CN104568537A (en) * | 2014-11-05 | 2015-04-29 | 华文蔚 | Method for treating biological micro-fluidic sample |
| CN105628660A (en) * | 2015-12-29 | 2016-06-01 | 大连理工大学 | Passive microvalve POCT chip |
| CN105628660B (en) * | 2015-12-29 | 2018-04-10 | 大连理工大学 | A kind of passive micro-valve POCT chips |
| CN108883414A (en) * | 2016-04-29 | 2018-11-23 | 建奥迪斯有限公司 | Method and component for molecule recycling |
| CN108883414B (en) * | 2016-04-29 | 2021-06-01 | 建奥迪斯有限公司 | Method and assembly for molecular recovery |
| US11135581B2 (en) | 2016-04-29 | 2021-10-05 | Creoptix Sa | Methods and assemblies for molecule recovery |
| CN110832325A (en) * | 2017-07-07 | 2020-02-21 | 救助科学技术有限责任公司 | Fluid analysis module and fluid analyzer |
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| CN100504389C (en) | 2009-06-24 |
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