CN103675306B - For automated analysis instrument and the system of micro-fluidic immune detection chip - Google Patents
For automated analysis instrument and the system of micro-fluidic immune detection chip Download PDFInfo
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- CN103675306B CN103675306B CN201310503132.1A CN201310503132A CN103675306B CN 103675306 B CN103675306 B CN 103675306B CN 201310503132 A CN201310503132 A CN 201310503132A CN 103675306 B CN103675306 B CN 103675306B
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- 238000001514 detection method Methods 0.000 title claims abstract description 50
- 238000004458 analytical method Methods 0.000 title claims abstract description 34
- 239000007788 liquid Substances 0.000 claims abstract description 44
- 239000002184 metal Substances 0.000 claims abstract description 34
- 229910052751 metal Inorganic materials 0.000 claims abstract description 34
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 25
- 230000033001 locomotion Effects 0.000 claims abstract description 22
- 238000000034 method Methods 0.000 claims abstract description 17
- 238000002347 injection Methods 0.000 claims abstract description 15
- 239000007924 injection Substances 0.000 claims abstract description 15
- 238000010521 absorption reaction Methods 0.000 claims abstract description 9
- 239000002699 waste material Substances 0.000 claims description 10
- 230000008569 process Effects 0.000 claims description 7
- 230000005540 biological transmission Effects 0.000 claims description 4
- MTZWHHIREPJPTG-UHFFFAOYSA-N phorone Chemical compound CC(C)=CC(=O)C=C(C)C MTZWHHIREPJPTG-UHFFFAOYSA-N 0.000 claims 1
- 238000002203 pretreatment Methods 0.000 abstract description 5
- 238000012545 processing Methods 0.000 abstract description 5
- 102000004169 proteins and genes Human genes 0.000 abstract description 4
- 108090000623 proteins and genes Proteins 0.000 abstract description 4
- 238000000746 purification Methods 0.000 abstract description 4
- 238000005070 sampling Methods 0.000 abstract description 4
- 238000000926 separation method Methods 0.000 abstract description 4
- 102000013529 alpha-Fetoproteins Human genes 0.000 description 6
- 108010026331 alpha-Fetoproteins Proteins 0.000 description 6
- 238000004140 cleaning Methods 0.000 description 4
- 238000013461 design Methods 0.000 description 4
- 239000012530 fluid Substances 0.000 description 4
- 238000003018 immunoassay Methods 0.000 description 4
- 230000008901 benefit Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000004907 flux Effects 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- MHAJPDPJQMAIIY-UHFFFAOYSA-N Hydrogen peroxide Chemical compound OO MHAJPDPJQMAIIY-UHFFFAOYSA-N 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 230000036039 immunity Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 210000001503 joint Anatomy 0.000 description 2
- 238000003032 molecular docking Methods 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
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- 239000008363 phosphate buffer Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000004065 semiconductor Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
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Abstract
The invention discloses a kind of automated analysis instrument and system for micro-fluidic immune detection chip, this automated analysis instrument comprises apparatus frame and the mechanical movement module, the liquid that are fixed on described apparatus frame pipette module and detection module; Described mechanical movement module comprises three-dimensional machinery arm and is fixed on the working metal pipe on described three-dimensional machinery arm, and the motion of described three-dimensional machinery arm drives described working metal pipe to move for assembling and the card of suction nozzle; Described liquid pipettes module and is communicated with absorption and the injection of described working metal pipe for liquid; Described detection module reads and transmits for chemiluminescence signal on micro-fluidic immune detection chip. This instrument connects computer and forms system, round Realization sample pre-treatments, target protein separation and purification, sample and reagent injection and sucking-off, chip clean, the organic combination of sampling and analysis of signal processing, make microflow control technique automation, microminiaturization, more accurate and easier.
Description
Technical field
The present invention relates to microfluidic chip system technical field, relate in particular to a kind of for micro-fluidic immunityAutomated analysis instrument and the system of detection chip.
Background technology
Microfluidic Immunoassay method is the technology that new development is in recent years got up, and the method is with analytical chemistryBe main, in conjunction with the achievement of the related disciplines such as biochemistry, physical chemistry and immunology, in micron order structureManipulation is received and is risen to skin and rises volume fluid, carries out immune response detection, has that volume is little, specific area large, insteadShort between seasonable, analysis speed fast, reagent and amount of samples is few and the many indexs of Multi-example such as detect at the advantage simultaneously.Discovery and disease early diagnosis to diseases related mark in clinical disease proteomics research have importantPractical value. Conventional immunoassay needs long analysis time, and liquid handling process is also cumbersome,Flux smaller (at every turn can only detect one or several samples), and need many antibody reagents.Microfluidic analysis chip can overcome these shortcomings effectively. For example, considerably less in analyzing samples amountWhen sample, microflow control technique shows extremely strong advantage, and conventionally needing sample size is that the experiment of several milliliters is being adoptedWith after microflow control technique, only need the sample size of several microlitres, greatly save the consumption of sample and reagent.And the combination of microfluidic analysis technology and immunoassay, can also overcome tradition immunity to a certain extentOther shortcoming of analyzing, therefore, has caused researcher's extensive concern in recent years. Microflow control technique applicationIn the greatly raising of Promote immunity detection technique of immune detection, be with a wide range of applications with importantUsing value.
But, in current Microfluidic Immunoassay method, except the step that can complete on chip, itsHe needs to have come by hand mostly operation (as the detection of reading of sample pre-treatments, introducing, cleaning and signal)Become, complex steps, had both increased workload, also easily occurred detecting error, cannot be applied in easily realityTime detection on the spot in, be not easy to extensive use and the popularization of micro-fluidic detection method. And body completelyExisting micro-fluidic this is that high flux detects the advantage of face in this in essence. Therefore, be necessary to develop oneKind of full-automatic micro-fluidic detection analytical instrument, round Realization sample pre-treatments, target protein separation and purification,Sample and reagent injection and sucking-off, chip clean, the organic combination of sampling and analysis of signal processing, make miniflowControl techniques automation, microminiaturization, more accurate and easier.
Summary of the invention
The object of the present invention is to provide a kind of automated analysis instrument for micro-fluidic immune detection chip andSystem, this system round Realization sample pre-treatments, target protein separation and purification, sample and reagent inject withSucking-off, chip clean, the organic combination of sampling and analysis of signal processing, make microflow control technique automation, micro-Type, more accurate and easier.
For realizing object of the present invention, provide following technical scheme:
For an automated analysis instrument for micro-fluidic immune detection chip, comprise apparatus frame and be fixed onMechanical movement module on described apparatus frame, liquid pipette module and detection module; Described mechanical movement mouldPiece comprises three-dimensional machinery arm and is fixed on the working metal pipe on described three-dimensional machinery arm, described three-dimensional machinery armMotion drive described working metal pipe to move for the assembling of suction nozzle and card; Described liquid pipettes module and connectsLogical described working metal pipe is for absorption and the injection of liquid; Described detection module is for micro-fluidic immune detectionOn chip, chemiluminescence signal reads and transmits.
Preferably, described mechanical movement module also comprises workbench, on described workbench, arrange chip groove,Reagent trough and suction nozzle groove, described chip groove is used for placing micro-fluidic immune detection chip, and described reagent trough is used forPlace reagent bottle, described suction nozzle groove is used for placing suction nozzle and assembles for working metal pipe.
Preferably, described chip groove, reagent trough and suction nozzle groove are arranged side by side successively, and after described suction nozzle grooveFace arranges waste liquid tank.
Preferably, the edge of described waste liquid tank arranges suction nozzle draw-in groove, falls for the card of suction nozzle on working metal pipe.
Preferably, described liquid pipette module comprise stepper motor and described step motor control syringe pump andConduit, described syringe pump is communicated with described working metal pipe by conduit.
Preferably, described detection module is CCD camera, and described CCD camera is fixed on described three-dimensional machineryArm, moves with the motion of described three-dimensional machinery arm.
Preferably, described three-dimensional machinery arm by three in X, Y and Z-direction cover servomotor, leading screw andGuide rail forms.
Preferably, described apparatus frame is light closed structure.
Preferably, described apparatus frame is mainly made up of metal framework and metallic plate.
Preferably, described CCD camera connects computer, and described computer receives and process described CCD phaseThe chemiluminescence signal of machine transmission.
For an automatic analysis system for micro-fluidic immune detection chip, comprise automation as aboveThe computer of analytical instrument and connection thereof.
Beneficial effect of the present invention is: the automated analysis instrument for micro-fluidic immune detection chip of the present inventionDevice, comprise apparatus frame and be fixed on mechanical movement module, liquid on described apparatus frame pipette module andDetection module; Described mechanical movement module comprises three-dimensional machinery arm and is fixed on the gold on described three-dimensional machinery armBelong to instrumentation tubes, the motion of described three-dimensional machinery arm drive described working metal pipe to move for the assembling of suction nozzle andCard falls; Described liquid pipettes module and is communicated with absorption and the injection of described working metal pipe for liquid; Described inspectionSurvey module reading and transmitting for chemiluminescence signal on micro-fluidic immune detection chip. Above-mentioned automatic partitionAnalyse instrument and connect computer formation automatic analysis system, can carry out miniflow to micro-fluidic immune detection chipThe aligning of keyholed back plate road injection port, the position of reagent are fixed, the absorption of liquid and get, chemiluminescence signalRead and analytic function, round Realization sample pre-treatments, target protein separation and purification, sample and reagent noteEnter with sucking-off, chip to clean, the organic combination of sampling and analysis of signal processing, make microflow control technique automation,Microminiaturized, more accurate and easier. Use this automatic analysis system can realize high flux and detect, operationSimply, workload is little.
Brief description of the drawings
Fig. 1 is that the automated analysis instrument primary structure for micro-fluidic immune detection chip of the present invention is overlookedFigure, wherein, 1 represents three-dimensional machinery arm, and 2 represent working metal pipe, and 3 represent workbench, and 4 represent CCDCamera, 31 represent chip groove, and 32 represent reagent trough, and 33 represent suction nozzle groove, and 34 represent waste liquid tank, 35 tablesShow suction nozzle draw-in groove.
Fig. 2 is the automated analysis instrument primary structure side-looking for micro-fluidic immune detection chip of the present inventionFigure, wherein, 1 represents three-dimensional machinery arm, and 2 represent working metal pipe, and 3 represent workbench, and 4 represent CCDCamera, 35 represent suction nozzle draw-in groove.
Fig. 3 is that the result reading for the automatic analysis system of micro-fluidic immune detection chip of the present invention is shoneSheet, wherein white square represents chemiluminescence signal, the brighter expression chemiluminescence signal of white square is stronger.
Detailed description of the invention
Below by detailed description of the invention, embodiment of the present invention are described in detail. Art technology peopleMember will be understood that, below embodiment be only the preferred embodiment of the present invention so that better understandInvention, thereby should not be considered as limiting scope of the present invention. For a person skilled in the art, the present inventionCan there be various modifications and variations, within the spirit and principles in the present invention all, any amendment of doing, etc.With replacement or improvement etc., within all should being included in protection scope of the present invention.
Please refer to Fig. 1 and Fig. 2, for the automatic analysis system of micro-fluidic immune detection chip, mainly by instrumentDevice framework, mechanical movement module, liquid pipette module, detection module and computer (not shown) groupClose and form. Wherein, apparatus frame fixing for each module position of instrument; Mechanical movement module is for goldThe three-dimensional that belongs to the position of instrumentation tubes moves, and on working metal pipe, assembling and the card of suction nozzle falls; Liquid pipettesModule is for absorption and the injection of micro liquid; Detection module is for chemiluminescence signal on micro-fluidic chipRead, and by image transmission to computer. Realized mechanical movement module, liquid pipettes module, inspectionThe effective integration of surveying module and computer, this system meets the requirement of carrying out immune detection at micro-fluidic chip,It is a kind of novel full-automatic microfluidic chip system.
Below in conjunction with Fig. 1 and Fig. 2, each assembly of this automatic analysis system is described in detail.
Apparatus frame is mainly made up of metal framework and metallic plate. Due to the chemiluminescence signal of micro-fluidic chipIn detection, need lucifuge, this apparatus frame is designed to light closed structure, is designed with and can opens in the front side of instrumentDoor (not shown) for the picking and placeing of chip and reagent, the agent structure mechanical movement mould of systemPiece is fixed on this apparatus frame.
Mechanical movement module mainly comprises three-dimensional machinery arm 1, working metal pipe 2 and workbench 3. Three-dimensional machineTool arm 1 is by the cover of three in X, Y and Z-direction servomotor, leading screw and guide rail (not shown) structureBecome. The bottom structure of working metal pipe 2 is according to suction nozzle apex structure design, can with suction nozzle tight connecting,The top of working metal pipe 2 and syringe pump and conduit are tightly connected, and reach the air-tightness requirement of liquid line.Chip groove 31, reagent trough 32, suction nozzle groove 33 and waste liquid tank 34 are set, described chip groove on workbench 331 for placing micro-fluidic immune detection chip, and described reagent trough 32 is for placing reagent bottle, described suction nozzle groove33 for placing suction nozzle for the assembling of working metal pipe, and described waste liquid tank 34 is for splendid attire system waste liquid. InstituteThe edge of stating waste liquid tank 34 arranges suction nozzle draw-in groove 35, according to the design of suction nozzle top end diameter, for working metal pipe 2The card of upper suction nozzle falls.
Liquid pipettes module and is made up of syringe pump and the conduit of step motor control, and described syringe pump passes through conduitBe communicated with described working metal pipe 2, can complete accurate absorption and the injection of 3-100 μ L liquid.
Detection module is CCD(Charge-coupledDevice, charge coupled cell) camera 4, also canBeing called ccd image sensor, is a kind of semiconductor devices, optical image can be converted into data signal,Complete the imaging of chemiluminescence signal on micro-fluidic chip and be transferred to computer. In addition, any for miniflowThe reading of chemiluminescence signal on control chip, and image transmission all be can be used as to this to the camera of computerThe detection module of invention.
This operation principle and process that is used for the automatic analysis system of micro-fluidic immune detection chip is: by micro-Fluidic chip is placed in chip groove 31, and the reagent using in test is placed in reagent trough 32, and suction nozzle is placed in suction nozzle groove 33In for subsequent use, start instrument, write fill order after relative program. Under programme-control, three-dimensional machinery arm 1Working metal pipe 2 is moved to directly over suction nozzle behind position, and Z axis moves downward, with suction nozzle closed butt joint, logicalThe motion of crossing three-dimensional machinery arm 1 moves to working metal pipe 2 directly over reagent bottle, and Z axis moves downward, willSuction nozzle on working metal pipe 2 bottoms immerses below the liquid level of reagent bottle, draws quantitative liquid by syringe pumpAfter, move to micro-fluidic chip injection port directly over, Z axis moves downward, and realizes suction nozzle and micro-fluidic chipThe closed butt joint of injection port, injects quantitative liquid by syringe pump to micro-fluidic chip, and this process is onceLiquid assimilating and injection action, by the orderly repetition of this process, can realize micro-fluidic chip immune detectionWhole fluid operated work. Micro-fluidic chip is designed with waste liquid pool, can effectively collect and store waste liquid.After immune detection reaction finishes, three-dimensional machinery arm 1 moves to CCD camera 4 directly over chip reaction zone,CCD camera 4 changes the chemiluminescence signal on chip into electrical signal, and then is transferred to computer and carries outData processing and analysis.
Through test, this system has three higher axles and moves precision, can realize suction nozzle and micro-fluidic coreThe accurate docking of sheet injection port, and the docking of working metal pipe 2 and suction nozzle, mobile accuracy is 0.01mm;This system has high liquid control precision, can realize accurate absorption and the injection of 3 μ L-100 μ L liquid,The error of moving liquid precision is ± 5%; This system has compared with high detection sensitivity, can realize on micro-fluidic chipEfficiently reading of chemiluminescence signal, more than the time for exposure can reach 50min; This system has higherAutomation control ability, after coding, only needs a key, just can realize the behaviour of whole immune detection testsDo; This system simple and compact for structure, volume is little, easy to use.
This system also has following characteristics: adopt suction nozzle to complete liquid and pipette action, avoid mutual pollution, economizeRemove cleaning step; Adopt high-precision servo motor, effectively control the three-dimensional position of working metal bar; Adopt highAccuracy CCD camera imaging, colleges and universities catch the chemiluminescence signal on micro-fluidic chip; Reasonable in design, realizeMechanical movement module, liquid pipette optimizing and combining of module and detection module, realized the integrated of system,Small volume, is convenient to detect operation; Full-automatic design, easy and simple to handle, realize micro-fluidic chip immunityDetect the automatic operation of test, only need a key, complete whole operating process.
Use the automatic analysis system for micro-fluidic immune detection chip of the present invention to detect micro-fluidic immunityThe reacted chemiluminescence signal of detection chip. Particularly, in 1,2 directions of micro-fluidic immune detection chipOn fix in advance alpha-fetoprotein (AFP) coated antibody, in A, B, C and tetra-micro-fluidic pipelines of D, comply withInferior pass into confining liquid (bSA etc.), AFP solution, cleaning fluid (phosphate buffer etc.),AFP detects antibody, cleaning fluid and chemical luminescence for liquid (hydrogen peroxide and luminol etc.), wherein A, B, CWith the concentration of the AFP solution adding in D pipe be followed successively by 200ng/mL, 100ng/mL, 50ng/mL and25ng/mL. The result photo (Fig. 3) reading shows: AFP solution concentration is higher, and chemiluminescence signal moreBy force, white square is brighter.
Applicant's statement, the present invention illustrates detailed features of the present invention and square in detail by above-described embodimentMethod, but the present invention is not limited to above-mentioned detailed features and method detailed, does not mean that the present invention is necessaryRelying on above-mentioned detailed features and method detailed could implement. Person of ordinary skill in the field should understand,Any improvement in the present invention, selects the equivalence replacement of component and the interpolation of auxiliary element, concrete to the present inventionThe selections of mode etc., within all dropping on protection scope of the present invention and open scope.
Claims (6)
1. for an automated analysis instrument for micro-fluidic immune detection chip, it is characterized in that, comprise instrumentDevice framework and the mechanical movement module, the liquid that are fixed on described apparatus frame pipette module and detection module;Described mechanical movement module comprises three-dimensional machinery arm and is fixed on the working metal pipe on described three-dimensional machinery arm,The motion of described three-dimensional machinery arm drives described working metal pipe to move for assembling and the card of suction nozzle; DescribedLiquid pipettes module and is communicated with absorption and the injection of described working metal pipe for liquid; Described detection module is used forOn micro-fluidic immune detection chip, chemiluminescence signal reads and transmits;
Wherein, described mechanical movement module also comprises workbench, on described workbench, arrange chip groove,Reagent trough and suction nozzle groove, described chip groove is used for placing micro-fluidic immune detection chip, and described reagent trough is used forPlace reagent bottle, described suction nozzle groove is used for placing suction nozzle and assembles for working metal pipe;
Described liquid pipettes syringe pump and the conduit that module comprises stepper motor and described step motor control, instituteState syringe pump and be communicated with described working metal pipe by conduit;
Described three-dimensional machinery arm is made up of the cover of three in X, Y and Z-direction servomotor, leading screw and guide rail;
Described chip groove, reagent trough and suction nozzle groove are arranged side by side successively, and after described suction nozzle groove, arrange uselessLiquid bath; The edge of described waste liquid tank arranges suction nozzle draw-in groove, falls for the card of suction nozzle on working metal pipe;
Described liquid pipettes module can complete accurate absorption and the injection of 3-100 μ L liquid; Described automationIt is 0.01mm that three axles of analytical instrument move precision.
2. automated analysis instrument according to claim 1, is characterized in that, described detection module isCCD camera, described CCD camera is fixed on described three-dimensional machinery arm, with the motion of described three-dimensional machinery armAnd move.
3. automated analysis instrument according to claim 1, is characterized in that, described apparatus frame isLight closed structure.
4. automated analysis instrument according to claim 1, is characterized in that, described apparatus frame masterTo be formed by metal framework and metallic plate.
5. automated analysis instrument according to claim 2, is characterized in that, described CCD cameraConnect computer, described computer receives and processes the chemiluminescence signal of described CCD camera transmission.
6. for an automatic analysis system for micro-fluidic immune detection chip, comprise as claim 1-5Automated analysis instrument described in any one and the computer of connection thereof.
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| CN105203746B (en) * | 2015-10-26 | 2017-07-14 | 深圳华迈兴微医疗科技有限公司 | A kind of POCT chemiluminescence immunoassay systems and its analysis method |
| CN108704677A (en) * | 2018-04-27 | 2018-10-26 | 广州万孚生物技术股份有限公司 | A kind of micro-fluidic chip and the analytical instrument containing it |
| CN113188848A (en) * | 2021-04-12 | 2021-07-30 | 攸太科技(台州)有限公司 | Urine tracking method |
| CN117101749B (en) * | 2023-10-20 | 2024-01-30 | 江苏集萃中科纳米科技有限公司 | Microfluidic chip fixed sample adding device and sample adding method |
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| CN201707339U (en) * | 2010-04-26 | 2011-01-12 | 常州鸿臻电子技术有限公司 | Full automatic testing center |
| CN102906573A (en) * | 2010-02-23 | 2013-01-30 | 瑞昂尼克公司 | Self-contained biological assay apparatus, methods, and applications |
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| US20040099334A1 (en) * | 2002-11-21 | 2004-05-27 | Gilson, Inc. | Liquid handling apparatus |
| US20040241872A1 (en) * | 2003-03-17 | 2004-12-02 | Qiagen Operon, Inc. | Optical detection liquid handling robot system |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102906573A (en) * | 2010-02-23 | 2013-01-30 | 瑞昂尼克公司 | Self-contained biological assay apparatus, methods, and applications |
| CN201707339U (en) * | 2010-04-26 | 2011-01-12 | 常州鸿臻电子技术有限公司 | Full automatic testing center |
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