CN101942385A - Full-automatic hybridization oven and implementation method and application thereof - Google Patents
Full-automatic hybridization oven and implementation method and application thereof Download PDFInfo
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- CN101942385A CN101942385A CN 200910040857 CN200910040857A CN101942385A CN 101942385 A CN101942385 A CN 101942385A CN 200910040857 CN200910040857 CN 200910040857 CN 200910040857 A CN200910040857 A CN 200910040857A CN 101942385 A CN101942385 A CN 101942385A
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Abstract
The invention provides a full-automatic hybridization oven and an implementation method and application thereof. The full-automatic hybridization oven structurally comprises a hybridization reacting box, a sample feeding device for adding a reagent to the hybridization reacting box, a reagent supply device, a reagent temperature controlling device and a control terminal, wherein, the hybridization reacting box is provided with hybridization membrane strips, the reagent supply device is sequentially connected with the reagent temperature controlling device and the sample feeding device through a pipeline, and the control terminal is connected with the reagent temperature controlling device. The hybridization process of the hybridization oven of the invention is controlled by the control terminal to realize full-automatic hybridization operation. The invention can be applied to existence and quantification of affinity mates in chemical molecular binding reactions such as nucleic acid hybridization, antibody-antigen reaction, receptor-ligand reaction, ion-culture medium reaction in a chemical binder and the like.
Description
Technical field
The present invention relates to molecular hybridization, be specifically related to a kind of fully automatic molecular hybridization instrument and its implementation and application.
Background technology
The complementary nucleotide sequence is called hybridization by the process that the Walson-Crick base pairing forms stable heterozygosis duplex molecule (dna molecular).This crossover process is a high degree of specificity, can carry out specific target sequence according to employed probe known array and detect.The making nucleic acid molecular hybridization technology has the specificity of very high sensitivity and height, thereby in biology field, this technology has been widely used in the qualitative and quantitative analysis of specific gene sequence in the making, genome of screening, the restriction enzyme mapping of clone gene and the aspects such as diagnosis of disease.In addition, it not only is used widely in biology field, and the application in clinical diagnosis also increases day by day.
The making nucleic acid molecular hybridization technology is different according to testing goal and detection means, can be divided into location (original position) hybridization in solution hybridization, solid-phase hybridization and the cell.Solid-phase hybridization can be divided into dot hybridization again, the gel electrophoresis trace shifts hybridization etc.Dot hybridization comprises forward dot hybridization and reverse dot blot hybridization again.
Dot hybridization is meant directly to be put DNA or RNA sample on nitrocellulose filter or nylon membrane, hybridizes with nucleic acid probe molecules then, whether to have special DNA or RNA in the show sample.And (the reverrsedotblot of reverse dot blot hybridization wherein, RDB) be a kind of dot hybridization technology that Saiki etc. proposes, this technology shifts hybridization with forward dot hybridization and gel electrophoresis trace and compares, have quick, easy, hypersensitivity, strong characteristics of property specially, especially gene type, detection in Gene Mutation, there is its special advantages aspects such as the detection of pathogenic agent.
The principle of work of reverse dot blot hybridization is to utilize the primer of marks such as vitamin H, and the specific amplified target DNA fragment is hybridized with specific probe then.It is different from general dot hybridization, and general dot blotting hybridization is that target DNA is fixed on nitrocellulose filter or the nylon membrane, the probe of mark and target DNA hybridization colour developing.Adopt this method to hybridize reflection at every turn and can only judge all whether DNA to be measured contains the homologous sequence of a certain probe, may contain tens to tens allelotrope (as HLA DRB site or thalassemia mutator gene etc.) for some locus, will be very numerous and diverse with this dot blotting hybridization method, even be difficult to accomplish.And RDB has solved this difficult problem just.RDB puts stand-by probe on nitrocellulose filter or the nylon membrane respectively earlier, point of each probe also compiles Shang number, (generally be product with dna sample to be measured again through the PCR specific amplification, carry out biotin labeling in advance at PCR primer 5 ' end, make the amplified production respective markers that vitamin H be arranged) hybridization with it, like this, sample to be checked will combine with the probe with homologous sequence, remove unconjugated dna sample through washing, because the marker that dna sample to be measured has the vitamin H class, combine the marker that just has the vitamin H class on the probe points of DNA to be measured, just can show hybridization signal through corresponding color reaction again.So once just can judge most of or whole allelotrope of a certain locus, therefore, utilize such principle of work can also be applied to gene type, pathogen detection, tumor research etc.
The RDB technology be with multiple probe stationary on same film, the sample DNA that participate in to detect does not simultaneously disturb again mutually, so the disposable examination of energy go out multiple different sequence, rather than once only can detect a unknown nucleotide sequence as traditional hybrid method.Because the multiple probe on same film is hybridized with different amplification PCR products simultaneously, and the probe that requires all to be incorporated on the film should have roughly the same Tm value, therefore, designed probe Tm value changed in more among a small circle.The RDB technology has also reduced false positive rate and false negative rate, and operational safety, simple, quick, and the film bar vegetation time is shorter, only needs several hours, can prepare in advance in a large number that to be put in 4 ℃ of preservations stand-by.After the pcr amplification product purpose fragment, only need hybridize, wash film (realized washing sameization of film condition, need not difference and wash the film condition), colour developing, judgement as a result etc.The somatotype that both had been applicable to a small amount of sample detects, and somatotype in the time of also applicable to a large amount of sample has the characteristics of susceptibility height, specificity height, good stability.
But traditional hybridization technique or anti-phase dot hybridization technology are all carried out in Glass tubing, centrifuge tube or plastics bag in proper temperature, and length not only consuming time, complicated operation are loaded down with trivial details, and poor repeatability as a result, influence the accuracy of results of hybridization.Existing Hybridization Oven mainly contains following two kinds: a kind of is water-bath, and water-bath is a kind of maintenance homothermic equipment, all needs manual operations in the whole process of carrying out hybridization, low, the poor temperature uniformity of temperature-controlled precision, and crossed contamination easily takes place; Another kind is triumphant general hybridization instrument, and triumphant general hybridization instrument has just increased automatic draining device with respect to water-bath, and many processes still need manual operations, belong to semi-automatic product, and temperature-controlled precision is not high, poor temperature uniformity, also than being easier to occur crossed contamination.
Summary of the invention
The objective of the invention is to overcome the shortcoming and defect of above-mentioned prior art, a kind of fully automatic molecular hybridization instrument is provided, hybridization instrument of the present invention has been realized full automatic hybridization operation.
Another object of the present invention is to provide the implementation method of above-mentioned fully automatic molecular hybridization instrument.
A further object of the present invention is to provide the application of above-mentioned fully automatic molecular hybridization instrument.
Purpose of the present invention is achieved through the following technical solutions: the fully automatic molecular hybridization instrument comprises: the hybridization box, be used for reagent is added to sample adding device, agent delivery device, reagent temperature regulating device and the control terminal of hybridization box; Described hybridization box is provided with the Hybond membrane bar, and described agent delivery device is connected with reagent temperature regulating device, sample adding device successively by pipeline, and described control terminal is connected with the reagent temperature regulating device.Like this, the agent delivery device is sent to the reagent temperature regulating device by the road with reagent, the reagent temperature regulating device carries out temperature control to reagent under the control of control terminal, the reagent temperature regulating device is sent to sample adding device to reagent more by the road, reagent is added to the hybridization box and central Hybond membrane bar carries out hybridization by sample adding device.
In order to realize locational accurate application of sample better, fully automatic molecular hybridization instrument of the present invention can also be provided with position transducer and be used to drive the transmission mechanism that sample adding device moves, described transmission mechanism is connected with control terminal, and described position transducer is fixed in sample adding device and is connected with control terminal.Like this, by position transducer the positional information of sample adding device is sent to control terminal, move by the positional information control actuator drives sample adding device of control terminal again according to described sample adding device, by sample adding device the hybridization box is carried out the reagent application of sample, thereby realized that from the position sample adding device carries out the accurate control of application of sample to the hybridization box.
Described hybridization box has a plurality of, described sample adding device is provided with the liquid feeding head that is used for reagent is joined the hybridization box, described reagent temperature regulating device comprises spiral heater, described agent delivery device comprises the reagent abstraction module, described liquid feeding head, spiral heater are identical with the quantity of reagent abstraction module, each reagent abstraction module all is connected with 1 spiral heater, 1 liquid feeding head successively by pipeline, and each liquid feeding head is all fixing to carry out application of sample to one or more hybridization boxes.Like this, each spiral heater is set working parameters such as Heating temperature independently by control terminal, thereby the reagent that the reagent abstraction module transmits is carried out independently temperature control, and then the reagent after the temperature control is fixedly joined in one or more hybridization boxes through the liquid feeding head.Thereby hybridization in each hybridization box has all been realized independently thermostatic control, and, because each Hybond membrane bar all is arranged on 1 hybridization box independently, avoided hybridization in the different hybridization boxes possibility of crossed contamination to occur.
Described spiral heater comprises spiral tube, heating rod, temperature sensor, heat-conducting silicone grease and shell, be filled with heat-conducting silicone grease between heating rod and the spiral tube, described spiral tube, heating rod, temperature sensor and heat-conducting silicone grease all are packaged in the shell, and described temperature sensor all is connected with control terminal with heating rod.Like this, temperature sensor sends to control terminal with the temperature information of heat-conducting silicone grease, by the working parameters such as Heating temperature of control terminal according to this temperature information setting heating rod, heating rod carries out temperature control by the heat-conducting silicone grease with good heat conductive performance to the reagent in the spiral tube.
Described reagent abstraction module comprises pump and stepper-motor, and the stepper-motor driving pump carries out the extraction of reagent, and reagent is sent to by the road the spiral tube of spiral heater.
Described sample adding device is used for liquid discharge head that the reacted waste liquid of hybridization box is discharged to be provided with also the time, and at this moment, the full-automatic hybridization instrument of the present invention is correspondingly provided with pumping equipment, and liquid discharge head is connected by pipeline with pumping equipment.
Described pumping equipment comprises pump and stepper-motor, and described pump is connected with the liquid discharge head of sample adding device by pipeline.After in the hybridization box, finishing hybridization, the positional information that control terminal transmits according to the position transducer that is fixed in sample adding device, the liquid discharge head of control actuator drives sample adding device is discharged reacted waste liquid in the hybridization box through pumping equipment.
This present invention fully automatic molecular hybridization instrument also is provided with the washing unit that is used to clean liquid discharge head, comprises the toilet seat that is positioned at sample adding device below, is used for the toilet seat water supply installation that supplies water to toilet seat, and the toilet seat water supply installation is connected with toilet seat by pipeline.Concrete, described toilet seat water supply installation comprises pump and stepper-motor, by the stepper-motor driving pump toilet seat is supplied water, and with after the liquid discharge head effluent discharge, liquid discharge head is cleaned.
For the reagent in the enabling hybridization reaction box and Hybond membrane bar can react more fully, fully automatic molecular hybridization instrument of the present invention can also further be provided with and rock device, this rocks device and comprises shaker platform and be used to drive the micromotor that shaker platform is rocked, described hybridization box is installed on the shaker platform, micromotor is connected with shaker platform, rocks together to drive shaker platform and hybridization box.More specifically optimal technical scheme is: described shaker platform comprises that hole slot and bracing frame are provided with hole slot, described hybridization box is placed on the hole slot of shaker platform, described micromotor is connected with shaker platform by rotating shaft, micromotor drives shaker platform through rotating shaft and rocks, and also rocks together thereby drive the hybridization box that is placed in the hole slot.
Described micromotor can also be connected with control terminal, by the switch and the shaking speed of control terminal control micromotor, thereby realizes control to the switch and the shaking speed of hybridization box by driving shaker platform.
Preferably, the hole slot in described hybridization box and the described shaker platform all adopts the rectangle structure that matches.
Temperature is constant when keeping hybridization better, and fully automatic molecular hybridization instrument of the present invention can also be provided with automatic water-bath temperature regulating device, and the water-bath temperature regulating device comprises tank, temperature sensor and heating rod automatically.The bottom of described hybridization box is positioned at below the upper edge of tank, with the part of enabling hybridization reaction box not in the water of tank.Described temperature sensor and heating rod all are positioned at tank, and all are connected with control terminal.Like this, by temperature sensor the temperature information of water in the tank is sent to control terminal, control terminal is controlled heating rod work according to the temperature information of water in this tank, thereby reaches the temperature control to water in the tank, makes the water in the tank keep constant temperature.
Preferably, described automatic water-bath temperature regulating device can also comprise the internal circulation system of the water internal recycle that is used for realizing tank.
Preferably, described automatic water-bath temperature regulating device can also comprise and be used for tank is supplied water and the water drainage-supply system of draining, this water drainage-supply system is provided with the water level sensor that is connected with control terminal, tank is being supplied water or during draining, by water level sensor the water level information of tank is sent to control terminal, control terminal according to the water level information of this tank to water supply and sewage operation control.
Hybond membrane bar in the described hybridization box is preferably hybridization probe film bar.
The implementation method of above-mentioned fully automatic molecular hybridization instrument comprises the steps:
(1) each hybridization box that will be provided with the Hybond membrane bar is placed in the hole slot of the shaker platform of rocking device;
(2) inject an amount of water by water drainage-supply system toward tank, control automatic water-bath temperature regulating device, make the water in the tank keep constant temperature, guarantee that each hybridization box is not all partly in the water of tank by control terminal;
(3) reagent is sent to the reagent temperature regulating device by the road by the agent delivery device, by the work of control terminal control reagent temperature regulating device, makes reagent temperature regulating device output constant temperature reagent to sample adding device;
(4) sample adding device carries out the reagent application of sample to each hybridization box under the drive of transmission mechanism, makes the Hybond membrane bar in reagent and the hybridization box 1 carry out hybridization.
In the aforesaid method, the positional information of the position transducer of sample adding device is fixed in the control terminal collection, the motion of transmission mechanism is controlled, thereby realized the locational application of sample control of hybridization box.
In the aforesaid method, rock device under the control of control terminal, the hybridization box that drives in the hole slot rocks reagent in the enabling hybridization reaction box and Hybond membrane bar sufficient reacting.
In the aforesaid method, control terminal can carry out fault and show and error reporting, comprise that monitoring is fixed in position transducer, transmission mechanism, the heating rod of spiral heater, the temperature sensor of spiral heater, the micromotor that rocks device and the shaker platform of sample adding device, automatically temperature sensor, the working order of each parts of these fully automatic moleculars such as water level sensor hybridization instrument of the heating rod of water-bath temperature regulating device and water drainage-supply system automatically of water-bath temperature regulating device, when operation irregularity occurring, carry out fault and show and error reporting.
Whether and quantitatively the existence that fully automatic molecular hybridization instrument of the present invention can be applicable to affinity spouse in ion-chemical molecular association reactions such as Pei radical reaction in nucleic acid hybridization, antibody-antigen-reactive, receptor-ligand reaction, the chemical combination.
The present invention has following advantage and effect with respect to prior art:
(1) fully automatic molecular hybridization instrument of the present invention extracts reagent by the agent delivery device, control the reagent temperature regulating device by control terminal then reagent is carried out temperature control, make the reagent that adds in the hybridization box keep constant temperature, by sample adding device reagent is added in the hybridization box again and hybridize with the Hybond membrane bar, whole crossover process is finished by machine, realized full-automatic crossover operation, and, consuming time less, simple to operate, good reproducibility and accurately as a result, the temperature-controlled precision height helps industry and applies;
(2) in fully automatic molecular hybridization instrument of the present invention, each Hybond membrane bar all is arranged on 1 hybridization box independently, has avoided hybridization in the different hybridization boxes possibility of crossed contamination to occur;
(3) fully automatic molecular hybridization instrument of the present invention can also be provided with position transducer and transmission mechanism, thereby has realized that from the position sample adding device carries out the accurate control of application of sample to the hybridization box;
(4) the reagent temperature regulating device in the fully automatic molecular hybridization instrument of the present invention adopts spiral heater, can realize the temperature control to reagent better;
(5) fully automatic molecular hybridization instrument of the present invention is provided with pumping equipment, and sample adding device is provided with liquid discharge head, can be automatically, easily reacted waste liquid in the hybridization box is discharged through pumping equipment;
(6) fully automatic molecular hybridization instrument of the present invention rocks on the device by the hybridization box is placed in, rocking device by control terminal control rocks the hybridization box, improved the concussion uniformity coefficient of reagent and Hybond membrane bar, reagent in the enabling hybridization reaction box and Hybond membrane bar can react fully, can improve speed of response effectively, and control terminal can be controlled the shaking speed that rocks device, thereby realizes the control to hybridization box shaking speed;
(7) fully automatic molecular hybridization instrument of the present invention is by being provided with automatic water-bath temperature regulating device, under control terminal control, automatically the water-bath temperature regulating device makes the water in the tank be in constant temperature, with the hybridization box partly not in water, can also be provided with the internal circulation system of the water internal recycle that is used for realizing tank, guarantee the constant of hybridization temperature better;
(8) fully automatic molecular hybridization instrument of the present invention can be applicable to affinity spouse in ion-chemical molecular association reactions such as Pei radical reaction in nucleic acid hybridization, antibody-antigen-reactive, receptor-ligand reaction, the chemical combination existence whether and quantitatively.
Description of drawings
Fig. 1 is the front view of present embodiment fully automatic molecular hybridization instrument;
Fig. 2 is the rear view of present embodiment fully automatic molecular hybridization instrument;
Fig. 3 is the structural representation of transmission mechanism and sample adding device;
Fig. 4 is the structural representation that rocks device, tank and water-bath temperature controller;
Fig. 5 is the structure explosive view that rocks device, tank and water-bath temperature controller;
Fig. 6 is the structural representation of spiral heater;
Fig. 7 is the water drainage-supply system structure iron;
Fig. 8 is the fundamental diagram of this enforcement fully automatic molecular hybridization instrument.
Embodiment
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.
Embodiment
As shown in figs. 1 and 3, the fully automatic molecular hybridization instrument in the present embodiment comprises: a plurality of hybridization boxes 1, be used for reagent be added to the hybridization box sample adding device 2, agent delivery device, reagent temperature regulating device, rock device 3, automatically water-bath temperature regulating device, pumping equipment, position transducer 4, be used to drive transmission mechanism 5 and the control terminal 6 that sample adding device 2 moves.Wherein, described control terminal is provided with display screen 4.
Described hybridization box 1 is provided with the Hybond membrane bar, and described Hybond membrane bar is preferably hybridization probe film bar.
Described agent delivery device is connected with reagent temperature regulating device, sample adding device 2 successively by pipeline, and described control terminal 6 is connected with the reagent temperature regulating device.Like this, the agent delivery device is sent to the reagent temperature regulating device by the road with reagent, the reagent temperature regulating device carries out temperature control to reagent under the control of control terminal 6, the reagent temperature regulating device is sent to sample adding device 2 to reagent more by the road, by sample adding device 2 reagent is added to hybridization box 1 and carries out hybridization with central Hybond membrane bar.
As shown in Figure 3, described sample adding device 2 is provided with 6 liquid feeding heads 75 that are used for reagent is joined hybridization box 1, and 21 liquid discharge head 86-1 and liquid discharge head 8-2 that are used for the 1 reacted waste liquid discharge of hybridization box.
As illustrated in fig. 1 and 2, described pumping equipment comprises that pump 7 and stepper-motor 8 have 2, and pump 9-1 and stepper-motor 10-1 form 1 pumping equipment, and pump 9-2 and stepper-motor 10-2 form another pumping equipment., pump 9-17 is connected with the liquid discharge head 8-16 of sample adding device 2 by pipeline.Pump 9-2 is connected with the liquid discharge head 8-2 of sample adding device 2 by pipeline.After in hybridization box 1, finishing hybridization, the positional information that control terminal 6 transmits according to the position transducer 4 that is fixed in sample adding device 2, liquid discharge head 8-1, liquid discharge head 8-26 that control transmission mechanism 5 drives sample adding device 2 discharge reacted waste liquid in the hybridization box 1 through pumping equipment.
As shown in Figure 1, also be provided with the toilet seat 9 that is used to clean liquid discharge head 6 under the described sample adding device 2, toilet seat 9 is connected with pump 10 by pipeline, is supplied water by 10 pairs of toilet seats of stepper-motor 11 driving pumps 9, with after liquid discharge head 6 effluent discharges, liquid discharge head 6 is cleaned.
As shown in Figure 2, described reagent temperature regulating device comprises 6 spiral heaters 1112, and each spiral heater 1112 all is connected with control terminal 6.Each spiral heater 1112 is set working parameters such as Heating temperature independently by control terminal 6, thereby the reagent that reagent extraction supply module device is transmitted carries out independently temperature control, and then the reagent after the temperature control fixedly joined in one or more hybridization boxes 1 through liquid feeding head 75, thereby hybridization in each hybridization box 1 has all been realized independently thermostatic control, and, because each Hybond membrane bar all is arranged in 1 hybridization box 1 independently, avoided hybridization in the different hybridization boxes 1 possibility of crossed contamination to occur.
As shown in Figure 6, described each spiral heater 1112 includes spiral tube 1213, heating rod 1314, temperature sensor, heat-conducting silicone grease and shell, be filled with heat-conducting silicone grease between heating rod 1314 and the spiral tube 1213, described spiral tube 1213, heating rod 1314, temperature sensor and heat-conducting silicone grease all are packaged in the shell, and described temperature sensor all is connected with control terminal 6 with heating rod 1314.Like this, temperature sensor sends to control terminal 6 with the temperature information of heat-conducting silicone grease, by the working parameters such as Heating temperature of control terminal 6 according to this temperature information setting heating rod 1314, heating rod 1314 carries out temperature control by the heat-conducting silicone grease with good heat conductive performance to the reagent in the spiral tube 1213.
As illustrated in fig. 1 and 2, described agent delivery device comprises 6 reagent abstraction modules, each reagent abstraction module includes 1 pump 1415 and stepper-motor 1516, stepper-motor 1516 driving pumps 1415 carry out the extraction of reagent, and reagent is sent to by the road the spiral tube 1213 of 1 spiral heater 1112, then, this spiral heater 1112 is connected with 1 liquid feeding head 75 by pipeline, and each liquid feeding head 75 is all fixing to carry out application of sample to one or more hybridization boxes 1.
As shown in Figure 1, described position transducer 4 is fixed in sample adding device 2 and is connected with control terminal 6.Described transmission mechanism 5 is connected with control terminal 6, and transmission mechanism 5 also is connected with sample adding device 2.As shown in Figure 3, transmission mechanism 5 comprises line slideway 1617 and drag chain 1718.Like this, by position transducer 4 positional information of sample adding device 2 is sent to control terminal 6, move by the positional information control transmission mechanism 5 driving sample adding devices 2 of control terminal 6 again according to described sample adding device 2,75 pairs of hybridization boxes of liquid feeding head 1 by sample adding device 2 carry out the reagent application of sample, thereby have realized that from the position 2 pairs of hybridization boxes of sample adding device 1 carry out the accurate control of application of sample.
For the reagent in the enabling hybridization reaction box 1 and Hybond membrane bar can react more fully, present embodiment fully automatic molecular hybridization instrument further is provided with and rocks device 3.As Fig. 4 or with shown in 5,
The described device 3 that rocks comprises shaker platform 19, is used to drive micromotor 1820, left rotary shaft 1921 and the right spindle 2022 that shaker platform 19 is rocked, described shaker platform 19 is provided with hole slot 23, described hybridization box 1 is placed on the hole slot 23 of shaker platform 19 and comprises hole slot 21 and bracing frame 22, described hole slot 21 matches to merge with hybridization box 1 and is placed on the bracing frame 22, described shaker platform 19 bracing frames 22 are driven by left rotary shaft 1921 and right spindle 2022 and rock, and are placed in also rocking with hybridization box 1 in the hole slot 2123 thereby drive.
Hole slot 2123 in described hybridization box 1 and the described shaker platform 19 all adopts the rectangle structure that matches, and, described hybridization box 1 and hole slot 2123 equal consistence are set to a plurality of, like this, can carry out a plurality of hybridizations simultaneously by respectively laying 1 hybridization box 1 at a plurality of hole slots 2123.
Described micromotor 1820 is connected with control terminal 6, by the switch and the shaking speed of control terminal 6 control micromotors 1820, thereby realizes control to the switch and the shaking speed of hybridization box 1 by driving shaker platform 19.
As Fig. 4 or with shown in 5, temperature is constant when keeping hybridization better, present embodiment fully automatic molecular hybridization instrument is provided with automatic water-bath temperature regulating device, and the water-bath temperature regulating device comprises tank 2324, temperature sensor 2425 and heating rod 2526-1 and heating rod 2526-2 automatically.The bottom of described hybridization box 1 is positioned at below the upper edge of tank 2324, with the part of enabling hybridization reaction box 1 not in the water of tank 2324.Described temperature sensor 2425, heating rod 2526-1 and heating rod 2526-2 all are positioned at tank 124, and all are connected with control terminal 6.Like this, by temperature sensor 2425 temperature information of water in the tank 2324 is sent to control terminal 6, control terminal 6 is controlled heating rod 2526-1 and heating rod 2526-2 work according to the temperature information of water in this tank 2324, thereby reach temperature control, make the water in the tank 24 keep constant temperature water in the tank 2324.
Described automatic water-bath temperature regulating device also is provided with the internal circulation system of the water internal recycle that is used for realizing tank 2324.As Fig. 2,4 or and shown in 5, this internal circulation system comprises pump 27, pipeline 2628-1 and pipeline 2628-2 and pump 27, the two ends of pump 27 are connected with pipeline 2628-1, pipeline 2628-2 respectively.
As described in Figure 7, automatically the water-bath temperature regulating device also comprises and is used for tank 2324 is supplied water and the water drainage-supply system of draining.This water drainage-supply system is provided with the water level sensor 2829 that is connected with control terminal 6, and the end 2930 of supplying water is connected with pump 3031 by pipeline, and pump 31 is by stepper-motor 32 drivings, as illustrated in fig. 1 and 2.
As described in Figure 7,, as shown in Figure 2.Described water level sensor 28 is used for tank 24 being supplied water or during draining, described water level sensor 29 is sent to control terminal 6 with the water level information of tank 2324, control terminal 6 according to the water level information of this tank 2324 to water supply and sewage operation control.During water supply, the valve 3233 of the end 30 that supplies water is opened, and is drawn water and is entered tank 2324 through the end 2930 that supplies water by stepper-motor 32 driving pumps 3031, and at this moment, the valve 35 of discharge ends 34 is closed;
During draining, the valve 3233 of the end 30 that supplies water is closed, and the valve 35 of discharge ends 34 is opened, and the water in the tank 2324 flows out automatically through discharge ends 3134.
Above-mentioned fully automatic molecular hybridization instrument is realized the method for molecular hybridization, as shown in Figure 8, comprises the steps:
(1) each hybridization box 1 that will be provided with the Hybond membrane bar is placed on the hole slot 2123 of described shaker platform 19 of rocking device 3;
(2) inject an amount of water by water drainage-supply system toward tank 2324, guarantee that each hybridization box 1 is not all partly in the water of tank 2324, and, need guarantee that also temperature sensor 2425, heating rod 2526-1 and heating rod 2526-2 be not in the water of tank 2324, gather the temperature information of water in the tanks 2324 and control heating rod 2526-1 and heating rod 2526-2 carries out work through temperature sensor 2425 by control terminal 6, make the water in the tank 2324 keep constant temperature;
(3) each pump 1415 of 6 in the agent delivery device is under the driving of the individual stepper-motor 1516 of 6 correspondences, reagent is extracted and is sent to by the road in the spiral heater 1112 of 6 correspondences, each spiral heater 1112 is under the control of control terminal 6, reagent is carried out temperature control, be transferred to 6 each liquid feeding heads 75 of sample adding device 2 then by pipeline;
(4) liquid feeding head 75 is added to reagent in the hybridization box 1, makes the Hybond membrane bar in reagent and the hybridization box 1 carry out hybridization.
In the aforesaid method, the temperature of water is preferably and is consistent in control of the temperature of 1112 pairs of reagent of each spiral heater and the tank 24.The control of the temperature of 1112 pairs of reagent of each spiral heater, generally remain on about 40 degree or below, and the temperature of water generally remains on 40 degree in the tank 24.
In the aforesaid method, when the Hybond membrane bar in reagent and hybridization box 1 carried out hybridization, internal circulation system started, and carries out internal recycle constantly to keep the water in the tank 2324, improved the homogeneity of the water heating in the tank 2324.
In the aforesaid method, the positional information that control terminal 6 is gathered the position transducer 4 that is fixed in sample adding device 2 is controlled the motion of transmission mechanism 5, thereby is made the application of sample process of 75 pairs of hybridization boxes 1 of liquid feeding head realize accurate position control.
In the aforesaid method, when the Hybond membrane bar in reagent and hybridization box 1 carries out hybridization, rock the switch and the shaking speed of the micromotor 1820 of device 3 by control terminal 6 controls, thereby realize to the switch of the hybridization box 1 in the hole slot 2123 and the control of shaking speed reagent in the enabling hybridization reaction box 1 and even, the sufficient reacting of Hybond membrane bar concussion by driving shaker platform 19.
In the aforesaid method, control terminal 6 can carry out fault by display screen 4 and show and error reporting, comprise and monitor the position transducer 4 that is fixed in sample adding device 2, transmission mechanism 5, the heating rod 1314 of spiral heater 1112, the temperature sensor of spiral heater 12, rock the micromotor 1820 and the shaker platform 19 of device 3, the temperature sensor 2425 of automatic water-bath temperature regulating device, the heating rod 2526-1 and the 2526-2 of automatic water-bath temperature regulating device, and the water level sensor 2829 of water drainage-supply system waits the working order of each parts of this fully automatic molecular hybridization instrument, when operation irregularity occurring, carry out fault by display screen 4 and show and error reporting.
Whether and quantitatively the existence that the utility model fully automatic molecular hybridization instrument can be applicable to affinity spouse in ion-chemical molecular association reactions such as Pei radical reaction in nucleic acid hybridization, antibody-antigen-reactive, receptor-ligand reaction, the chemical combination.
The foregoing description is a preferred implementation of the present invention; but embodiments of the present invention are not limited by the examples; other any do not deviate from change, the modification done under spirit of the present invention and the principle, substitutes, combination, simplify; all should be the substitute mode of equivalence, be included within protection scope of the present invention.
Claims (10)
1. the fully automatic molecular hybridization instrument is characterized in that, comprising: the hybridization box, be used for reagent is added to sample adding device, agent delivery device, reagent temperature regulating device and the control terminal of hybridization box; Described hybridization box is provided with the Hybond membrane bar, and described agent delivery device is connected with reagent temperature regulating device, sample adding device successively by pipeline, and described control terminal is connected with the reagent temperature regulating device.
2. fully automatic molecular hybridization instrument according to claim 1, it is characterized in that: also be provided with position transducer and be used to drive the transmission mechanism that sample adding device moves, described transmission mechanism is connected with control terminal, transmission mechanism also is connected with sample adding device, and described position transducer is fixed in sample adding device and is connected with control terminal.
3. fully automatic molecular hybridization instrument according to claim 1 and 2, it is characterized in that: also be provided with and rock device, this rocks device and comprises shaker platform and be used to drive the micromotor that shaker platform is rocked, described hybridization box is installed on the shaker platform, micromotor is connected with shaker platform, and described micromotor is connected with control terminal.
4. fully automatic molecular hybridization instrument according to claim 3, it is characterized in that: also be provided with pumping equipment, described sample adding device also is provided with the liquid discharge head that is used for the reacted waste liquid discharge of hybridization box, and liquid discharge head is connected by pipeline with pumping equipment.
5. fully automatic molecular hybridization instrument according to claim 4, it is characterized in that: also be provided with the washing unit that is used to clean liquid discharge head, comprise the toilet seat that is positioned at sample adding device below, be used for the toilet seat water supply installation that supplies water to toilet seat, the toilet seat water supply installation is connected with toilet seat by pipeline.
6. fully automatic molecular hybridization instrument according to claim 5, it is characterized in that: also be provided with automatic water-bath temperature regulating device, automatically the water-bath temperature regulating device comprises tank, temperature sensor and heating rod, the bottom of described hybridization box is positioned at below the upper edge of tank, with the part of enabling hybridization reaction box not in the water of tank, described temperature sensor and heating rod all are positioned at tank, and all are connected with control terminal.
7. fully automatic molecular hybridization instrument according to claim 6 is characterized in that: described automatic water-bath temperature regulating device can also comprise the internal circulation system of the water internal recycle that is used for realizing tank.
8. according to claim 6 or 7 described fully automatic molecular hybridization instruments, it is characterized in that: described automatic water-bath temperature regulating device can also comprise and be used for tank is supplied water and the water drainage-supply system of draining that this water drainage-supply system is provided with the water level sensor that is connected with control terminal.
9. the implementation method of fully automatic molecular hybridization instrument according to claim 8 is characterized in that, comprises the steps:
(1) each hybridization box that will be provided with the Hybond membrane bar is placed in the hole slot of the shaker platform of rocking device;
(2) inject an amount of water by water drainage-supply system toward tank, control automatic water-bath temperature regulating device, make the water in the tank keep constant temperature, guarantee that each hybridization box is not all partly in the water of tank by control terminal;
(3) reagent is sent to the reagent temperature regulating device by the road by the agent delivery device, by the work of control terminal control reagent temperature regulating device, makes reagent temperature regulating device output constant temperature reagent to sample adding device;
(4) sample adding device carries out the reagent application of sample to each hybridization box under the drive of transmission mechanism, makes the Hybond membrane bar in reagent and the hybridization box 1 carry out hybridization, when hybridizing reflection, rocks device drive hybridization box 1 and rocks.
10. according to the application of each described fully automatic molecular hybridization instrument of claim 1 to 8, it is characterized in that: be applied to the ion-Pei radical reaction in nucleic acid hybridization, antibody-antigen-reactive, receptor-ligand reaction, the chemical combination.
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| CN102533525A (en) * | 2011-12-22 | 2012-07-04 | 亚能生物技术（深圳）有限公司 | Full-automatic hybridization appliance |
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