A kind of micro-fluidic chemiluminescent analyzerTechnical field
The utility model is related to microfluidic arts, specifically, are related to a kind of micro-fluidic chemiluminescent analyzer.
Background technology
Chemiluminescence immune assay (Chemiluminescence immunoassay, CLIA) has high sensitivity, specialThe features such as property is strong, the range of linearity is wide, easy to operate, is widely used in tumor marker, infectious disease, endocrine function, hormone etc.The diagnosis of aspect, the chemiluminescence market share is increasing in recent years, and gradually having substituted enzyme linked immunological (ELISA) becomes immuneThe mainstream of diagnosis, detection content cover tumor markers, cardiac marker, thyroid gland energy, insulin, diabetes, infectious diseaseDisease, cell factor, hormone, allergic reaction and therapeutic drug monitori etc..
Chemistry In China illumination instrument includes semi-automatic Chemiluminescence Apparatus, Full-automatic chemiluminescence apparatus, portable Chemiluminescence Apparatus.Existing chemiluminescence detector is generally used for reading 96 holes or 48 hole microwell plates, and adding to last hole is loaded from the first holeThe sample time is longer, has a certain impact to the uniformity and accuracy of testing result.When required detection sample is less, one is used96 hole of monoblock or 48 hole microwell plates can cause greatly to waste.
Microfluidic chip technology (Microfluidics) is biological, chemical, medical analysis process sample preparation, insteadShould, separation, detection etc. basic operation units be integrated on the chip of one piece of micro-meter scale, be automatically performed analysis overall process.Due toIt in the great potential in the fields such as biology, chemistry, medicine, have been developed as a biology, chemistry, medicine, fluid, electronics,The brand-new research field of the subject crossings such as material, machinery.But the document for combining chemiluminescence and micro-fluidic chip is notIt is more, it is practical and can industrialization it is less.
Chinese patent 200910154432.7 discloses the micro-fluidic chip of capillary electrophoresis separation and chemiluminescence detection,Its flow passage structure is single, and sample introduction is not adequately mixed relatively low so as to cause reaction efficiency, is unable to reach maximum emission intensity.SomeGiant chemical luminaire detection sample size is big, easily obscures sample, and apparatus structure is complicated, bulky, cost is high, rightIn the small small hospital of sample size and community clinic and do not apply to.Some mini-plants only remain simple Mechanical course knotStructure, sensitivity and accuracy to pattern detection have a great impact.
In short, existing apparatus structure is complicated, volume is larger, real-time test is not suitable for（POCT）Application, practical applicationClinical detection convenience it is poor.Operating procedure is complicated, reagent adding, incubation, cleaning and etc. need manual operation, be easy to cause peopleTo slip up, the accuracy of testing result is influenced.Time-consuming for detection, and instrument work efficiency is low.
Utility model content
In order to solve drawbacks described above of the prior art, the utility model provides a kind of micro-fluidic chip for making instrument and placesConvenient, accurate positioning, testing result is accurately quick, reproducible micro-fluidic chemiluminescent analyzer, with reference to chemiluminescence skillWhen being detected, chip is read using bar code scanner for the high susceptibility of art and the high degree of specificity of immunological techniqueInformation determines personalization detection scheme；Spring assembly ensures to combine between integrality, chip and the instrument of micro-fluidic chip closedProperty；The flowing of level sensing column monitor sample at any time drives the opening and closing for stopping valve rod control valve device, and detection reaction controlling is existedThe reaction zone of chip；Sample, reaction reagent, luminous substrate of chip reaction zone etc. can be uniformly mixed by ultrasonic module；It is most laggardData are read in row optical detection.
To achieve the above object, the technical solution adopted in the utility model is：A kind of micro-fluidic chemiluminescent analyzer, bagFluid path gas path module, chip load-on module, ultrasonic module are included, fluid path gas path module, chip load-on module, ultrasonic module order are setIt puts, it is characterised in that：Chip load-on module is for chip placement and chip is driven to translate, and chip load-on module is clamped equipped with chipDevice；Fluid path gas path module include the first power part, the first power part control under synchronization lifting gas-blowing column, cleaning fluid column,Bottom fluid column, level sensing column；The only valve that fluid path gas path module further includes the second power part, lifted under the control of the second power partColumn；Gas-blowing column, cleaning fluid column, bottom fluid column, level sensing column, only the top of valve rod is equipped with elastic mechanism；Chip is located at reactionDuring position, gas-blowing column, cleaning fluid column, bottom fluid column, level sensing column bottom with chip surface flexible contact.
Gas-blowing column, cleaning fluid column, bottom fluid column are hollow structure and bottom is respectively inflatable mouth, cleaning solution mouth, bottom liquid mouth,When chip is located at response location, inflatable mouth, cleaning solution mouth, bottom liquid mouth with chip surface flexible contact.
Chip clamping device includes the chip slot for chip placement, and heating plate is equipped with below chip slot；Chip clampsDevice further includes the chip fixture block positioned at chip groove edge, the first elastic portion in chip fixture block and outside chip fixture blockLocating piece, the end of locating piece is fixed；First elastic portion one end connects chip fixture block, and the other end enters in the hole of locating piece simultaneouslyIt is connected with locating piece.
Ultrasonic transducer provides ultrasonic signal for ultrasonic module, is the prior art.The ultrasonic module includes the 3rdPower part, the eccentric rod being connected with the 3rd power part positioned at eccentric boom end and are located remotely from the connecting rod of one end of shaft, fixationUltrasonic head fixed block in connecting rod, the ultrasonic head stretched out upwards from ultrasonic head fixed block, ultrasonic head and chip surface flexible contact,Be equipped in ultrasonic head fixed block with ultrasonic head lower end against the 3rd elastic portion；The ultrasonic module is further included to be consolidated with ultrasonic headDetermine the ultrasonic synchronization blocks that block is fixedly connected, ultrasonic synchronization blocks are bonded setting with the 3rd line slideway, connecting rod also with ultrasonic synchronization blocksIt is fixedly connected；3rd power part causes eccentric rod to rotate and drives ultrasonic head fixed block, ultrasonic synchronization blocks, the synchronous rotary of connecting rodLifting, ultrasonic synchronization blocks can move on the 3rd line slideway.
The fluid path gas path module includes fluid path gas circuit fixed plate, and fluid path gas circuit fixed plate is fixedly connected with the air blowingColumn, cleaning fluid column, bottom fluid column, level sensing column, gas-blowing column clean fluid column, bottom fluid column, level sensing column parallel in orderRow, the first power part promote the lifting of fluid path gas circuit fixed plate；Elastic mechanism（That is spring）It is sheathed on gas-blowing column, cleaning fluid column, bottomThe top of fluid column, level sensing column；Level sensing post detection to sample will go into the valve position of chip internal when it is second dynamicThe promotion of power portion stops valve rod and moves down and push down the valve position of chip.
The fluid path gas path module is equipped with second straight line guide rail, and the second straight line guide rail is fixed and fluid path gasRoad fixed plate moves on second straight line guide rail.
Chip load-on module is additionally provided with first straight line guide rail, and gear rotates band carry-over bar and translates, and it is straight that rack fits in firstLine guide rail surface and rack is synchronized with the movement with first straight line guide rail, rack end connection chip clamping device.
The analyzer of the utility model is additionally provided with counter, and counter is connected with photomultiplier PMT, photomultiplierPMT lower ends are for being directed at chip upper surface, and when carrying out optical detection, chip is between light source and photomultiplier PMT.
A kind of application method of micro-fluidic chemiluminescent analyzer, which is characterized in that include the following steps：
（1）When starting detection, chip is put into chip loading device, chip is clamped by chip clamping device solidIt is fixed；Moving chip loading device makes chip enter instrument internal；
（2）When chip reaches response location, lead screw motor drives fluid path gas circuit fixed plate to be pressed downward, when gas circuit liquid circuit moduleStopping pushing when being pressed onto chip, independent spring assembly is arranged at gas-blowing column, cleaning fluid column, bottom fluid column, the top of level sensing column,Make inflatable mouth, cleaning solution mouth, bottom liquid mouth, level sensing column flexible contact chip；Blower pump outwards blows from inflatable mouth and by coreThe sample of piece adding mouth is blown into the reaction zone of chip internal, when level sensing post detection to sample will go into chip internalValve position, another lead screw motor of gas circuit liquid circuit module drive immediately only valve rod be pressed downward, push down the valve location on chip,So that sample does not continue to move along, while blower pump is stopped；
（3）The motor of ultrasonic module rotates, and the eccentric rod rotation being fixed on motor shaft, eccentric rod drives connecting rod rotation,Connecting rod connects ultrasonic head fixed block by axis pin, and interlinking lever end is also connected with ultrasonic synchronization blocks, and ultrasonic head fixed block is fixed on superIn sound synchronization blocks, ultrasonic synchronization blocks are fixed on line slideway, and ultrasonic head limiting slot is located in ultrasonic head fixed block and ultrasonic head/V is in ultrasonic head limiting slot and can be slided up and down in ultrasonic head limiting slot；When motor rotates, drive ultrasonic head upwardMovement, line slideway move upwards simultaneously；When ultrasonic head touches chip reaction zone, the spring assembly of ultrasonic head beneath makes to surpassSound head and chip flexible contact；Start the liquid in ultrasonic mixing reaction zone after contact fully, allow examination in sample and reaction zoneAgent is fully reacted；After the completion of ultrasonic mixing, ultrasonic module moves downward, and stops when reaching initial position；
（4）Cleaning pump injects cleaning solution, and cleaning solution flows to the reaction zone of chip internal from cleaning solution mouth, by cleaning for several timesAfterwards, substrate pump adds in substrate solution；Ultrasonic module moves upwards carry out mixing, and substrate and reagent is made fully to react；After the completion of reaction,Chip loading device moves backward, reaches optical detecting module, reads data, and chip exits after the completion of reading, and detection is completed.
In step（2）In, after chip reaches fixed position, bar code scanner reads the bar code on chip, reads coreThe information of on piece, determines whether loading chip matches with cuit, and report an error prompting if mismatching, and is carried out if matchingIt operates in next step.
Compared with prior art, the utility model has the following advantages：The utility model collection is multiple to be installed on one, can be withBarcode scanning is carried out to each chip, determines personalization detection project；Sample, reaction reagent, luminous substrate are filled using ultrasonic moduleDivide mixing, finally carry out optical detection, read sample data, make entirely to detect program standardization, automation, and detection sensitivityHigh, reliable results.For this equipment on the basis of every detection function is ensured, structure has carried out great simplification, small volume, systemMake it is at low cost, it is easy to operate, be suitble to carry out real-time test at one's side in patient, testing result can be quickly obtained.
Description of the drawings
Fig. 1 is the complete machine front view of the utility model；
Fig. 2 is the three-dimensional structure diagram of the utility model；
Fig. 3 is the structure diagram of chip load-on module；
Fig. 4 is chip load-on module chips fixture block, the structure diagram of chip slot；
Fig. 5 is the structure diagram of fluid path gas path module；
Fig. 6 is the structure diagram of ultrasonic module；
Fig. 7 is the spring-loaded plunger structure diagram of fluid path gas path module；
Fig. 8 is gas-blowing column, cleaning fluid column, bottom fluid column, the structure diagram of level sensing column；
Wherein, 1 counter, 2 fluid path gas path modules, 3 chip load-on modules, 4 ultrasonic generators, 5 ultrasonic modules, 6a topsCylinder, 6b step end faces, 6c lower parts cylinder, the first lead screw motors of 2a, the second lead screw motors of 2b, 2c bar code scanners, 2dTwo line slideways, 2e springs, 2f spring-loaded plungers, 3a first straight line guide rails, 3b racks, 3c chip slots, 3d chip fixture blocks, 3e firstElastic portion, 3f locating pieces, 3g heating plates, 5a ultrasound heads, 5b ultrasound synchronization blocks, the 3rd line slideways of 5c, 5d ultrasound head fixed blocks,5e ultrasound head limiting slots, 5f axis pins, 5g springs, 7 inflatable mouths, 8 cleaning solution mouths, 9 bottom liquid mouths, 10 level sensing columns, 11 stop valve rod,12 flange bearings, 13 connecting rods, 14 eccentric rods, 15 motors, 16 fluid path gas circuit fixed plates.
Specific embodiment
The utility model is further described with reference to embodiment.
According to Fig. 1-8, a kind of micro-fluidic chemiluminescent analyzer including fluid path gas path module 2, chip load-on module 3, surpassesSound module 5, fluid path gas path module, chip load-on module, ultrasonic module order are set, and chip load-on module is used for chip placement simultaneouslyChip translation is driven, chip load-on module is equipped with chip clamping device；Fluid path gas path module includes the first power part（Such as firstLead screw motor 2a）, the first power part control under synchronization lifting inflatable mouth 7（Positioned at air blowing column bottom）, cleaning solution mouth 8（PositionIn cleaning liquid column bottom）, bottom liquid mouth 9（Positioned at bottom liquid column bottom）, level sensing column 10；It is dynamic that fluid path gas path module further includes secondPower portion（Such as the second lead screw motor 2b）, the second power part control under lift only valve rod 11；Gas-blowing column, cleaning fluid column, bottomFluid column, level sensing column top are equipped with elastic mechanism（That is spring 2e）；Only valve rod top is equipped with elastic mechanism（That is spring-loaded plunger2f）；When chip is located at test position, inflatable mouth, cleaning solution mouth, bottom liquid mouth, level sensing column connect with chip upper surface flexibilityIt touches.Only valve rod is by stopping valve rod top（That is spring-loaded plunger）Only valve rod lower part（Also it is column structure）It is formed by connecting, spring-loaded plungerTop and the second lead screw motor nut are fixed on same fixed plate（This fixed plate is not fluid path gas circuit fixed plate）On.
According to Fig. 8, gas-blowing column, cleaning fluid column, bottom fluid column, level sensing column are by top cylinder 6a, lower part cylinder 6c groupsInto spring（It is not shown in FIG. 8）It is sleeved on the cylinder of top, lower part column diameter is more than top cylinder, top cylinder and lower partThe abutted surface of cylinder is step end face 6b, and spring top withstands on fluid path gas circuit fixed plate lower surface, and spring bottom end withstands on lower postStep end face at the top of body, top cylinder, lower part cylinder are that the hollow bulb of hollow structure and the two communicates.Fluid path gas circuit is fixedPlate and pillar step end face play position-limiting action.
Chip clamping device includes the chip slot 3c for chip placement；Chip clamping device is further included positioned at chip trough rimChip fixture block 3d, the first elastic portion 3e of edge and the locating piece 3f outside chip fixture block, the first elastic portion left end connection chipPosition to the right in fixture block, the first elastic portion right end are placed in the left end hole of locating piece and are connected with locating piece, the right side of locating pieceEnd is fixed（Fig. 3 and Fig. 4）.Before chip is put into, chip fixture block 3d left ends also cover a small amount of region of chip slot 3c, and chip is putAfter entering, chip fixture block 3d is moved to right, and chip fixture block and chip contact area are cambered surface（With the globoidal structure phase of the corner of chipMatch somebody with somebody）, due to the presence of elastic portion, chip is held out against by chip fixture block.
Chip load-on module is additionally provided with first straight line guide rail 3a, and gear rotates band carry-over bar and translates 3b, and rack fits in theOne straight line guide rail surface and rack is synchronized with the movement with first straight line guide rail, rack end connection chip clamping device.
Micro-fluidic chemiluminescent analyzer, is additionally provided with counter 1, and counter 1 connects with photomultiplier PMT and based onCalculate photon numbers, photomultiplier PMT（For optical detecting module）For being directed at chip upper surface, light source is arranged at chip for lower endBetween photomultiplier PMT.
The ultrasonic module includes the 3rd power part（For example it is motor 15）And the motor axis connection of the 3rd power partEccentric rod 14, the connecting rod 13 for being connected to eccentric boom end and being located remotely from one end of motor shaft, the ultrasonic head for being fixed on connecting rod are consolidatedDetermine block 5d, the ultrasonic head 5a stretched out upwards from ultrasonic head fixed block, ultrasonic head and chip lower surface reaction zone flexible contact, ultrasoundHead fixed block in be equipped with ultrasonic head lower end against the 3rd elastic portion（That is spring 5g）；The ultrasonic module is further included and surpassedThe ultrasonic synchronization blocks 5b that sound head fixed block is fixedly connected, ultrasonic synchronization blocks are bonded setting, small end with the 3rd line slideway 5cAlso it is fixedly connected with ultrasonic synchronization blocks；3rd power part eccentric rod is rotated and drive ultrasonic head fixed block, ultrasonic synchronization blocks,The synchronous rotary lifting of connecting rod, ultrasonic synchronization blocks move on the 3rd line slideway（Fig. 6）.
The fluid path gas path module includes fluid path gas circuit fixed plate 16, and fluid path gas circuit fixed plate is fixedly connected with described blowGas port, cleaning solution mouth, bottom liquid mouth, level sensing column, inflatable mouth, cleaning solution mouth, bottom liquid mouth, level sensing column, only valve rod is by suitableSequence is arranged in parallel, and the first power part promotes the lifting of fluid path gas circuit fixed plate；Elastic mechanism is placed on gas-blowing column, cleaning fluid column, bottom liquidThe top of column, level sensing column；Level sensing post detection to sample i.e. will go into the valve position of chip internal when the second powerPortion's promotion stops valve rod and moves down and push down the valve position of chip；Only valve rod top is spring-loaded plunger 2f（Spring-loaded plunger is an elasticityStructure, finished product）；The fluid path gas path module be equipped with second straight line guide rail 2d, the second straight line guide rail it is fixed andFluid path gas circuit fixed plate moves on second straight line guide rail（Fig. 5 and Fig. 7）.
Specific detection process is as follows：When starting detection, synchronous motor drives rack-and-pinion to move forward, while drives chipLoading device moves forward, and when chip loading device is moved out outside instrument, is put into chip, there is chip clamping in chip loading deviceDevice, chip clamping device can fix chip and make chip positioning.Chip is put well, and synchronous motor drives chip loadingDevice enters instrument internal.
Chip loading device reaches fixed position（That is response location）When, the bar code scanner 2c of top is read on chipBar code, read the information on chip, determine whether loading chip matches with cuit, report an error and carry if mismatchingIt wakes up, operation is carried out down-walk if matching.First lead screw motor drives fluid path gas circuit fixed plate to be pressed downward, when gas circuit fluid path fillsIt puts and stops pushing when being pressed onto chip, inflatable mouth, cleaning solution mouth, bottom liquid mouth, level sensing column are each to have independent spring aboveDevice ensures inflatable mouth, cleaning solution mouth, bottom liquid mouth, level sensing column flexible contact chip, and chip will not be destroyed, whileIt can guarantee that inflatable mouth, cleaning solution mouth, bottom liquid mouth, level sensing column can combine closely with chip, gas leakage will not occur, leakage is askedTopic.The pump of inflatable mouth is first started to work, and the sample of chip adding mouth is blown into the reaction zone of chip internal, when level sensing columnDetect that sample will go into the valve position of chip internal, another lead screw motor of gas circuit level gauge drives only valve immediatelyColumn is pressed downward, and pushes down the valving on chip so that sample will not continue to move forward, while blower pump is stopped.
The stepper motor of ultrasonic module rotates, the eccentric rod rotation being fixed on motor shaft, linkage by axis pin andEccentric rod is connected to instrument, is consolidated in connecting rod and axis pin with flange bearing 12, the other end of connecting rod by axis pin 5f and ultrasonic headDetermine block 5d to link together, ultrasonic head fixed block is fixed on ultrasonic synchronization blocks 5b, and ultrasonic synchronization blocks are fixed on the 3rd straight line and leadOn rail 5c, ultrasonic head limiting slot 5e is located in ultrasonic head fixed block, and ultrasonic head /V is in ultrasonic head limiting slot and can beIt is slided up and down in ultrasonic head limiting slot.When synchronous motor rotates, by more than mechanism ultrasonic head is driven to move upwards, the 3rd straight lineGuide rail moves upwards simultaneously, and the 3rd line slideway control ultrasonic module straight line moves up and down, while also controls ultrasonic module will notGeneration double swerve.When ultrasonic head 5a touches chip reaction zone, the spring assembly below ultrasonic head（That is spring 5g）, make itIt can be in close contact with chip, while ensure that ultrasonic head will not damage chip by pressure again.Start after contact fully in ultrasonic mixing reaction zoneLiquid, the reagent allowed in sample and reaction zone fully reacts.After the completion of ultrasonic mixing, ultrasonic module moves downward, and reaches justIt stops during beginning position.
At this moment cleaning pump starts to note cleaning solution, is cleaned, and after cleaning for several times, substrate pump adds in substrate solution.UltrasoundModule moves upwards carry out mixing, and substrate and reagent is allowed fully to react.After the completion of reaction, chip loading device moves backward, arrivesUp to optical detecting module, data are read out.Chip exits after the completion of reading, and detection is completed.
There is chip clamping device in the chip loading device of the utility model, can fix and be accurately positioned chip.MiniflowIt controls on chip with bar code, bar code scanner can read the detection project of chip, so as to be confirmed whether and cuitMatching, instrument can add in different amounts of cleaning solution and substrate solution according to the requirement of different detection projects, so as to personalized detectionDisparity items.Inflatable mouth, cleaning solution mouth, bottom liquid mouth, level sensing column are each to have independent spring assembly above, can guarantee coreThe seal combined between the integrality of piece, chip and instrument.Level sensing column can monitor the flowing of liquid in chip, work as sampleWhen reaching valve position, only valve rod closes valve, and control reaction carries out in the reaction zone of chip.The utility model is mixed using ultrasoundThe liquid of even reaction zone, and repeatedly, sample, reagent, cleaning solution, substrate solution etc. can be made uniformly mixed and fully reacted, it protectsDemonstrate,prove the accuracy of testing result.Last optical detecting module detection chip information, user can read data on instrument.
The utility model belongs to miniature instrument, portable, easy to operate, and testing result is reliable, sensitivity is good, reduces realityThe operating procedure of personnel is tested, while sample analysis efficiency is improved, is avoided that sample contamination and sample information are chaotic.It can lead toIt crosses heating plate to be incubated, all reactions carry out in micro-fluidic chip, no waste liquid discharge.The micro-fluidic chemiluminescent analyzerIt may be used in clinical detection, qualitative, quantitative foundation provided for clinical treatment.
The above is only the preferred embodiment of the utility model, it should be pointed out that：For the common skill of the artFor art personnel, on the premise of the utility model principle is not departed from, several improvements and modifications can also be made, these improve andRetouching also should be regarded as the scope of protection of the utility model.