Background technology
The biochip technology utility model is in the middle and later periods nineties in last century; Until today, remain advanced Protocols in Molecular Biology, this technology can realize high-throughput, fast, the detection of sensitive gene and expression thereof; Because its sport technique segment is many, be not easy to control; Certain degree of difficulty is arranged in application, and real in gene test fields such as Clinical Laboratory, the popularization gradually is just formally to begin later in 2004, just has faster in nearly 2 years to develop.At present, the biochip technology Application Areas mainly contains gene expression spectrum analysis, new gene discovery, transgenation and polymorphism analysis, genomic library mapping, medical diagnosis on disease and prediction, drug screening, gene sequencing etc.; In addition, gene chip all will be made major contribution at aspects such as agricultural, food supervision, environment protection, judicial expertises.In view of the great potential of gene chip and tempting prospect, gene chip has become the focus of various countries academia and industry member research and development.Gene chip is as a brand-new high-tech product, and its technology has reached molecular level and in widespread use gradually all over the world.It is the detection means of a kind of irreplaceable highly sensitive, high-throughput and high specific, has the very long product life cycle.
Gene chip is made up of sheet glass or nylon membrane and probe array fixed thereon; The two ultimate principle is similar, the complicated process of preparation of glass-chip, and testing process is loaded down with trivial details; Especially signal detection needs laser scanner; Directly caused its use cost high, particularly can not effectively have been promoted in the clinical detection, so its R&D direction mainly is to scientific research mechanism in market; The exploitation of film chip then has clear superiorities such as preparation is simple relatively, easy and simple to handle, with low cost, very helps the popularization in market, more can realize the industrialization of achievement in research quickly and efficiently.Evaluating objects according to the DNA chip; From the genes involved DB, choose the specific gene sequence as probe; Carry out probe sequence and topological design thereof according to the nucleotide sequence of being selected for use, requirement is to make the probe combinations that is designed reasonable to hybridization high specificity, the probe array of testing gene.On the gene chip basis; In conjunction with round pcr to a large amount of rapid amplifyings of testing sample; Contain the testing sample DNA product of respective markers thing and the probe on the gene chip after the amplification and carry out specific hybrid, hybridization signal is analyzed, easily target gene in the test sample.
The use of gene chip mainly comprises DNA extraction, PCR, hybridizes, washes film, 5 steps of colour developing, and DNA extraction is simple to operate, and PCR and hybridization step all can be realized through self-reacting device; And wash the trivial operations of film and development step; Need to consume great amount of manpower and time, and, cause the error or the mistake of human factor easily because operation steps is many; Influence the result of use and the result of gene chip, this just presses for the operation of automatic equipment instead of manual.
The utility model content
The technical problem that the utility model mainly solves is the trivial operations of washing film and development step in the gene chip use in the prior art; Need to consume great amount of manpower and time; And because operation steps is many; Be easy to generate the error or the mistake of human factor, influence the result and the result of use of gene chip, proposed a kind of full-automatic hybridization instrument.
For solving the problems of the technologies described above; The technical scheme that the utility model adopts is: a kind of full-automatic hybridization instrument is provided; Comprise hybridization platform, frame, backboard; Said frame and backboard are fixed in said hybridization platform, and said hybridization platform is provided with agent delivery device, reagent preheating unit, and said reagent preheating unit and agent delivery device are complementary; Said frame is provided with the hybridization box, be used for said agent delivery device reagent add reaction box sample adding device, be used to drive sample adding device and be with mechanism synchronously, be used to drive sample adding device and be with mechanism synchronously, be used to drive the hybridization box and be with mechanism synchronously along the reciprocating Y axle of Y direction along the reciprocating Z axle of Z-direction along the reciprocating X axle of X-direction; Be provided with the Hybond membrane bar in the said hybridization box, said agent delivery device is connected with said sample adding device through pipeline; Said backboard be provided be used for communicating by letter with upper computer and control the X axle synchronously with mechanism, Z axle synchronously with mechanism and Y axle synchronously with the main circuit board of mechanism kinematic, be used to drive the power amplifier wiring board of reagent preheating unit.
Wherein, the outside of said hybridization box is provided with the hybridization temperature control unit, and said hybridization temperature control unit is connected with said power amplifier wiring board.
Wherein, said hybridization temperature control unit comprises the heating tank cover plate that is positioned at above the said hybridization box, holds the heating tank of said hybridization box, heating tank base plate and the semiconductor chilling plate between heating tank and heating tank base plate; Said heating tank and said semiconductor chilling plate thermo-contact, said semiconductor chilling plate is electrically connected with said power amplifier wiring board.
Wherein, said heating tank plate outer side also is provided with axial fan; The side of said hybridizing box also is provided with the radiator fan that is fixed on the hybridization platform.
Wherein, said hybridization box is connected with the stepper-motor that is used to rock the hybridization box through drive shaft.
Wherein, said reagent preheating unit also is connected with TP, and said TP is connected with said main circuit board.
Wherein, also be provided with liquid feeding peristaltic pump, drawing liquid peristaltic pump and be used to control the SV of agent delivery device on the said backboard.
Wherein, said sample adding device is provided with syringe needle, and said syringe needle is connected with said pipeline.
Wherein, said X axle and Z axle are provided with and are respectively equipped with the transmitter that is used to respond to the sample adding device movement position, and said Y axle is provided with the transmitter that is used to respond to hybridization box movement position.
The beneficial effect of the utility model is: be different from the trivial operations of washing film and development step in the prior art in the gene chip use; Need to consume great amount of manpower and time; And because operation steps is many; Cause the error or the mistake of human factor easily; Influence the result of use and the result of gene chip; The utility model provides X axle in a kind of full-automatic hybridization instrument can make the mechanical manipulator sample adding device along the setting movement of X axle and Z-direction more accurately and can the enabling hybridization reaction box more accurate along the setting movement of Y direction with mechanism synchronously through the Y axle with mechanism synchronously with mechanism, Z axle synchronously, can make the bearing accuracy of sample adding device and hybridization box more accurate with motion of mechanism synchronously through utilizing main circuit board control X, Y, Z axle; Drive the reagent preheating unit through the power amplifier wiring board, can satisfy reagent and carry out the required preheating temperature of hybridization.Can automatically realize washing, hybridization in the gene chip operative technique through main circuit board and power amplifier wiring board to the control and the driving action of each device; Steps such as enzyme is hatched, colour developing; The artificial participation lacked; Can greatly reduce product cost, effectively promote " popular " of gene chip and application widely, can create huge economic benefit.
Embodiment
By the technology contents, the structural attitude that specify the utility model, realized purpose and effect, know clearly below in conjunction with embodiment and conjunction with figs. and give explanation.
See also Fig. 1; The invention provides a kind of full-automatic hybridization instrument; Comprise hybridization platform 1,frame 3,backboard 2, saidframe 3 is fixed in said hybridization platform 1 withbackboard 2, and said hybridization platform 1 is provided with agent delivery device 10; Comprise Aliquid reagent bottle 107, Bliquid reagent bottle 105, Cliquid reagent bottle 108, aluminium box Aliquid reagent bottle 106, clearwater reagent bottle 103,POD reagent bottle 102, wasteliquid reagent bottle 101, colour developingliquid reagent bottle 104; The reagent preheating unit comprises the outer courage of the B liquid heating that is used for preheating Bliquid reagent bottle 105reagent 1051, is used for the A liquid hot-plate 1061 of preheating Aliquid reagent bottle 107 reagent, and said reagent preheating unit and agent delivery device 10 are complementary; The said position relation that is meant A liquid hot-plate 1061 and Aliquid reagent bottle 107 in the reagent preheating unit that is complementary adapts; Be convenient to the preheating of A liquid, theouter courage 1051 of B liquid heating in the reagent preheating unit adapts with the position relation of Bliquid reagent bottle 105, is convenient to the preheating of B liquid; Saidframe 3 is provided with the hybridization box, be used for reagent add reaction box sample adding device, be used to drive sample adding device and be withmechanism 312 synchronously, be used to drive sample adding device and be withmechanism 311 synchronously, be used to drive the hybridization box and be withmechanism 313 synchronously along the reciprocating Y axle of Y direction along the reciprocating Z axle of Z-direction along the reciprocating X axle of X-direction; Be provided with the Hybond membrane bar in the said hybridization box; Said X axle is withmechanism 312 to be connected with that X shaft step motor, said Z axle are withmechanism 311 to be connected with the Z shaft step motor synchronously, said Y axle is withmechanism 313 to be connected with y-axis stepper motor synchronously synchronously, and said X shaft step motor, y-axis stepper motor and Z shaft step motor are controlled by main circuit board; Saidbackboard 2 be provided be used for communicating by letter with said upper computer and control the X axle synchronously withmechanism 312, Z axle synchronously withmechanism 311 and Y axle synchronously with the main circuit board of mechanism's 313 motions, be used to drive the power amplifier wiring board of reagent preheating unit; Said agent delivery device 10 is connected with said sample adding device through pipeline.Wherein X-direction is the direction of sample adding device inframe 3 crossbeam upper edge crossbeam move left and right, and said Y direction is meant that the hybridization box is shifted to or away from the backboard travel direction, said Z-direction is the direction perpendicular to hybridization platform 1.
In one embodiment; The outside of said hybridization box is provided with the hybridization temperature control unit that is used to control reagent react temperature in the hybridizing box; Said hybridization temperature control unit is connected with said power amplifier wiring board, and said hybridization temperature control unit comprises the heatingtank cover plate 301 that is positioned at above the said hybridization box, and said heating cover plate is provided with evenly distributed accurate liquid hole; Theheating tank 302 that holds said hybridization box; Heatingtank base plate 303, thesemiconductor chilling plate 304 betweenheating tank 302 and heatingtank base plate 303, saidsemiconductor chilling plate 304 is used to provide the temperature required of hybridization; Saidheating tank 302 and saidsemiconductor chilling plate 304 thermo-contacts, saidsemiconductor chilling plate 304 is electrically connected with said power amplifier wiring board.Said heatingtank base plate 303 outsides also are provided with theaxial fan 305 that is used to quicken to reduce fluid temperature in the control hybridization box; Be convenient to control the temperature of hybridization; Whensemiconductor chilling plate 304 is too high; The TR that heat radiation makes temperature maintenance allow at hybridization, said heatingtank base plate 303 outsides also are provided withradiator element 306; The side of said hybridizing box also is provided with the radiator fan 4 that is fixed on the hybridization platform 1, is mainly used in the heat radiation that hybridization finishes back hybridization box, can play the effect of quick heat radiating.Said hybridization box is connected with the stepper-motor that is used to rock the hybridization box through drive shaft, can improve the success ratio of hybridization.Said reagent preheating unit also is connected with TP; Said TP is connected with said main circuit board; Be the reagent before the preheating hybridization, and can control the temperature of preheating, prevent too high temperature damage in the reagent to carry out the gene fragment of hybridization.Also be provided with peristaltic pump on the saidbackboard 2 and be used to control the SV of agent delivery device 10; Said peristaltic pump comprises liquid feeding peristaltic pump and drawing liquid peristaltic pump; Be convenient to whole device and carry out liquid feeding or discharge opeing processing efficiently, said SV comprises A liquid electromagnetic valve, B liquid electromagnetic valve, C liquid electromagnetic valve, waste liquid SV, clear water SV, colour developing liquid electromagnetic valve, POD SV and aluminium box A liquid electromagnetic valve.Said sample adding device is provided with liquidfeeding syringe needle 3111 andimbibition syringe needle 3112, and said liquidfeeding syringe needle 3111 is communicated with corresponding pipeline respectively with imbibition syringe needle 3112.Said X axle and Z axle are provided with and are respectively equipped with the transmitter that is used to respond to the sample adding device movement position, and said Y axle is provided with the transmitter that is used to respond to hybridization box movement position, are convenient to the accurate location of sample adding device and hybridization box.
During work; The Z axle moves reciprocatingly on Z-direction with liquid feeding syringe needle and the imbibition syringe needle that mechanism drives on the sample adding device synchronously; The X axle moves reciprocatingly on X-direction with the liquid feeding syringe needle and the imbibition syringe needle of mechanism's drive sample adding device synchronously; The Y axle is with mechanism to drive the hybridization box synchronously and on Y direction, is moved reciprocatingly; When the position of hybridization box and sample adding device adapts, when promptly the syringe needle on the sample adding device was aimed at the accurate liquid hole of heating tank cover plate on the hybridization box, sample adding device carries out corresponding liquid feeding or imbibition is handled.
Upper computer can be controlled parameter time etc. in quantity and each step of reagent liquid measure, reaction box of adding; And through the RS232 serial ports; Send order and start lower computer work, lower computer comprises main circuit board and power amplifier wiring board, and main circuit board is responsible for the collection of preheating temperature of reagent and hybridization temperature; Control X axle synchronously withmechanism 312, Y axle synchronously withmechanism 313 and Z axle synchronously with the motion ofmechanism 311, control is used to rock the running of the stepper-motor of hybridization box; The power amplifier wiring board is used for providing reaction required temperature according to the order control reagent preheating unit of upper computer and hybridization temperature control unit.The temperature controlled process of hybridization is: TP passes to upper computer with real-time collecting temperature through lower computer; And in upper computer, accomplish temperature data and handle; After producing corresponding temperature signal after pid algorithm and the computing and being sent to the FET on the thermal module drive plate in the lower computer; Drivesemiconductor chilling plate 304 work, provide hybridization required temperature with this.
Working process is following:
Semiconductor chilling plate is started working simultaneously in S1, A liquid hot-plate, the B liquid outer courage of heating and the hybridization region, when the temperature of A liquid temperature probe, B liquid temperature probe reaches 51 ℃, and A liquid hot-plate outage stopping heating.When the hybridization platform temperature reaches 51 ℃, just begin to carry out next step action.
S2, the energising of aluminium box A liquid electromagnetic valve; Pipeline is through liquid feeding peristaltic pump conducting two minutes (inject required liquid earlier and be convenient to that individual lead is arranged) in advance; Sample adding device and hybridization box move to first position (guarantee that syringe needle accurately aim at the liquid hole) respectively then, and successively according to setting reaction box quantity on the upper computer, unit liquid measure (mL) begins to inject aluminium box A liquid to each test kit on the hybridization platform again; After last aluminium box A liquid electromagnetic valve and liquid feeding peristaltic pump all cut off the power supply, just begin to carry out next step action.
S3, hybridization on the platform rock the stepper-motor energising after, drive the hybridization box, make it begin to rock by 25 times/PM, crossing phase begins, and after 30 minutes, just begins to carry out next step action.
S4, hybridization box are got back to valley; B liquid electromagnetic valve energising, B liquid be through liquid feeding peristaltic pump conducting two minutes (inject required liquid earlier and be convenient to that individual lead is arranged) in advance, and sample adding device and hybridization box move to first position (guarantee that syringe needle accurately aim at the liquid hole) respectively then; Again successively according to setting reaction box quantity on the upper computer; Unit liquid measure (mL) begins to take out earlier waste liquid to each test kit on the hybridization platform, and B liquid is injected in the back, the outer courage outage stopping of last B liquid heating heating; After B liquid electromagnetic valve and liquid feeding peristaltic pump all cut off the power supply, just begin to carry out next step action.
S5, hybridization platform begin by 25 times/when PM rocked, the drawing liquid peristaltic pump also began action, injected POD liquid to A liquid, washed mem stage through after 5 minutes, just began to carry out next step action.
S6, hybridization region begin cooling, and cooling fan rotates (axial fan is used for making the liquid fast cooling), the energising of POD liquid electromagnetic valve; POD liquid was through liquid feeding peristaltic pump conducting two minutes, and sample adding device and hybridization box move to first position respectively then, more successively according to setting reaction box quantity on the upper computer; Unit liquid measure (mL) begins to take out earlier waste liquid to each test kit on the hybridization platform; POD liquid is injected in the back, after last D liquid electromagnetic valve and liquid feeding peristaltic pump all cut off the power supply, just begins to carry out next step action.
S7, hybridization platform begin the action of rocking by 25 times/PM, and POD is hatched the stage through after 30 minutes, just begins to carry out next step action.
S8, hybridization platform are got back to valley, the energising of A liquid electromagnetic valve, and A liquid was through liquid feeding peristaltic pump conducting two minutes; Sample adding device and hybridization box move to first position respectively then; Successively according to setting reaction box quantity on the upper computer, unit liquid measure (mL) begins to take out earlier waste liquid to each test kit on the hybridization platform again, and A liquid is injected in the back; After last A liquid electromagnetic valve and liquid feeding peristaltic pump 1 all cut off the power supply, just begin to carry out next step action.
S9, hybridization platform begin the action of rocking by 25 times/PM, and A liquid is washed mem stage through after 5 minutes, just begins to carry out next step action.
S10, hybridization platform are got back to valley, the energising of A liquid electromagnetic valve, and A liquid was through liquid feeding peristaltic pump conducting two minutes; Sample adding device and hybridization box move to first position respectively then; Successively according to setting reaction box quantity on the upper computer, unit liquid measure (mL) begins to take out earlier waste liquid to each test kit on the hybridization platform again, and A liquid is injected in the back; After last A liquid electromagnetic valve and liquid feeding peristaltic pump all cut off the power supply, just begin to carry out next step action.
S11, hybridization platform begin the action of rocking by 25 times/PM, and A liquid repeats to wash mem stage through after 5 minutes, just begins to carry out next step action.
S12, hybridization platform are got back to valley, the energising of C liquid electromagnetic valve, and C liquid was through liquid feeding peristaltic pump conducting two minutes; Sample adding device and hybridization box move to first position respectively then; Successively according to setting reaction box quantity on the upper computer, unit liquid measure (mL) begins to take out earlier waste liquid to each test kit on the hybridization platform again, and C liquid is injected in the back; After last C liquid electromagnetic valve and liquid feeding peristaltic pump all cut off the power supply, just begin to carry out next step action.
S13, hybridization platform begin to rock number of times by 25 times/PM, and C liquid is washed mem stage through after 2 minutes, just begins to carry out next step action.
S14, shut down to wait for that alarm sound is reminded the experimenter, wait the experimenter to prepare behind the colour developing liquid, continue following steps by continuing button.
S15, the energising of demonstration liquid electromagnetic valve; The liquid road was through liquid feeding peristaltic pump conducting two minutes, and sample adding device and hybridization box move to first position respectively then, more successively according to setting reaction box quantity on the upper computer; Unit liquid measure (mL) begins to take out earlier waste liquid to each test kit on the hybridization platform; The back inject to show liquid, show that at last liquid electromagnetic valve and liquid feeding peristaltic pump all cut off the power supply after, just begin to carry out next step action.
S16, shut down to wait for that the colour developing stage just begins to carry out next step action through after 15 minutes.
S17, the energising of clear water liquid electromagnetic valve; Clear water liquid was through liquid feeding peristaltic pump conducting two minutes, and sample adding device and hybridization box move to first position respectively then, more successively according to setting reaction box quantity on the upper computer; Unit liquid measure (mL) begins to take out earlier waste liquid to each test kit on the hybridization platform; Clear water liquid is injected in the back, after last clear water liquid F SV and liquid feeding peristaltic pump all cut off the power supply, just begins to carry out next step action.
S18, hybridization platform begin the action of rocking by 25 times/PM, the clear water stage begin always along thetime 2 minutes after, just begin to carry out next step action.
S19, shut down to wait for that alarm sound reminds the experimenter to accomplish experiment, press the hybridization conclusion button after, fan stops, and waits for that the experimenter takes hybridizing box away.
S20, wash pipe: machine is from employing the clear water pipe blow-through; Wash bottle: the experimenter manually cleans POD bottle, colour developing liquid bottle.
Fig. 2 is the results of hybridization figure of gene fragment of the present invention, and wherein real point is the successful zone of hybridization.
The present invention is different from the trivial operations of washing film and development step in the prior art in the gene chip use; Need to consume great amount of manpower and time; And because operation steps is many; Cause the error or the mistake of human factor easily; Influence the result of use and the result of gene chip; The invention provides a kind of full-automatic hybridization instrument, can make sample adding device more accurately and can the enabling hybridization reaction box more accurate with mechanism synchronously with mechanism synchronously with mechanism, Z axle synchronously along the setting movement of Y direction through the Y axle along the setting movement of X axle and Z-direction through the X axle, and through utilizing main circuit board control X, Y, Z axle can make the bearing accuracy of sample adding device and hybridization box more accurate with motion of mechanism synchronously; Drive the reagent preheating unit through the power amplifier wiring board, can satisfy the preheating temperature that reagent carries out hybridization.Can automatically realize washing, hybridization in the gene chip operative technique through main circuit board and power amplifier wiring board to the control and the driving action of each device; Steps such as enzyme is hatched, colour developing; The artificial participation lacked; Can greatly reduce product cost, effectively promote " popular " of gene chip and application widely, can create huge economic benefit.
The above is merely embodiments of the invention; Be not so limit claim of the present invention; Every equivalent structure or equivalent flow process conversion that utilizes specification sheets of the present invention and accompanying drawing content to be done; Or directly or indirectly be used in other relevant technical fields, all in like manner be included in the scope of patent protection of the present invention.