CN108998343A - A kind of generating device of digital pcr microlayer model - Google Patents
A kind of generating device of digital pcr microlayer model Download PDFInfo
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- CN108998343A CN108998343A CN201810828771.8A CN201810828771A CN108998343A CN 108998343 A CN108998343 A CN 108998343A CN 201810828771 A CN201810828771 A CN 201810828771A CN 108998343 A CN108998343 A CN 108998343A
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Classifications
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
- B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
- B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
- B01L3/502769—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements
- B01L3/502784—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by multiphase flow arrangements specially adapted for droplet or plug flow, e.g. digital microfluidics
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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- B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
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- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0433—Moving fluids with specific forces or mechanical means specific forces vibrational forces
- B01L2400/0439—Moving fluids with specific forces or mechanical means specific forces vibrational forces ultrasonic vibrations, vibrating piezo elements
Abstract
The invention discloses a kind of generating devices of digital pcr microlayer model, are related to digital pcr technical field.The device includes micro-fluidic chip, power device and supersonic generator, it is provided with gas adding mouth, liquid adding mouth on micro-fluidic chip and ultrasonic atomizatio cavity, gas sample and liquid sample pass through gas adding mouth respectively by preset ratio and liquid adding mouth enters in ultrasonic atomizatio cavity；Power device is connected to gas access and liquid inlet, and the flowing for gas sample and liquid sample provides power；Supersonic generator is arranged on the outside of ultrasonic atomizatio cavity, can excite the liquid sample in ultrasonic atomizatio cavity and be atomized into microlayer model.The device is based on ultrasonic atomizatio principle can form a large amount of microlayer models in a very short period of time, and generation speed is fast, and low energy consumption, and preparation facilities structure is simple, at low cost.
Description
Technical field
The present invention relates to digital pcr technical field more particularly to a kind of generating devices of digital pcr microlayer model.
Background technique
Carry out biological study and it is actually detected when, the amount of DNA sample to be measured is less sometimes or extracts from sample
It is preferable in order to obtain as a result, it is often necessary ro first carrying out polymerase chain to the amount of sample molecule when DNA molecular concentration to be measured is lower
It reacts (Polymerase Chain Reaction, PCR).
Based on the digital pcr that microflow control technique development generates, there is reaction volume more smaller than conventional PCR, faster
Reaction speed, lower system noise and higher sensitivity.Digital pcr technology is in the detection of denier sample of nucleic acid, complicated back
Rare mutation detection and the advantage that embodies of expression quantity fine difference identification aspect have been commonly recognized under scape, and it is in gene table
Up to research, microRNA research, genome copy numbers identification, the detection of cancer markers rare mutation, pathogenic microorganisms identification, turn
The broad prospect of application that all various aspects such as gene element identification, NGS sequencing library accurate quantification and result verification have has been subjected to
To more and more concerns.
Common digital pcr technology depends on Microfluidic droplet chip, silicon wafer microwell array, piezo jets currently on the market
With the modes such as PDMS array.
The operation of Microfluidic droplet chip needs to connect the equipment such as pump, complicated for operation；The formation and transport of microlayer model are to even
Continuous phase fluid requirements are higher, otherwise there is the risk of droplet rupture.
Micropore is processed on silicon wafer, this mode higher cost, and it is higher to micropore coherence request.
High-precision piezo jets equipment cost is higher, and there are cross contamination risks；
PDMS chip batch production technology is immature, needs to be always maintained at subnormal ambient, and this method flexibility is poor；And
PDMS chip specific heat is larger, and circulation time is increased in amplification procedure.
Therefore, a kind of generating device of digital pcr microlayer model is needed to solve the above problems.
Summary of the invention
The purpose of the present invention is to provide a kind of generating devices of digital pcr microlayer model, to realize the simple, high of microlayer model
Effect and preparation at low cost.
To achieve this purpose, the present invention adopts the following technical scheme:
A kind of generating device of digital pcr microlayer model, comprising:
Micro-fluidic chip is provided with gas adding mouth, liquid adding mouth and ultrasonic atomizatio cavity, gas sample and liquid
Body sample is passed through the gas adding mouth by preset ratio respectively and the liquid adding mouth enters in the ultrasonic atomizatio cavity；
Power device, is connected to the gas access and liquid inlet, and the power device can be the gas sample
The flowing of this and the liquid sample provides power；
Supersonic generator is arranged on the outside of the ultrasonic atomizatio cavity, and the supersonic generator can will be described
Liquid sample excitation in ultrasonic atomizatio cavity is atomized into microlayer model.
The depth of the preferred embodiment of generating device as digital pcr microlayer model, the middle part of the ultrasonic atomizatio cavity is big
In the arrival end of the ultrasonic atomizatio cavity and the depth of outlet end.
The preferred embodiment of generating device as digital pcr microlayer model, the arrival end and the outlet end pass through tiltedly
Face is connect with the middle part, and the inclined-plane can be avoided the formation of dead volume.
The preferred embodiment of generating device as digital pcr microlayer model, the micro-fluidic chip and the ultrasonic atomizatio chamber
Ultrasonic probe slot is provided at the corresponding position in the middle part of body, the ultrasonic probe slot can accommodate the supersonic generator
Ultrasonic probe.
The preferred embodiment of generating device as digital pcr microlayer model further includes gas-liquid mixed pipeline, the gas sample-adding
One end and the described one end of liquid adding mouth far from the power device far from the power device of mouth are mixed with the gas-liquid
Pipeline connection is closed, the other end of the gas-liquid mixed pipeline is connected to the ultrasonic atomizatio cavity.
The preferred embodiment of generating device as digital pcr microlayer model, the liquid adding mouth and the gas-liquid mixed pipe
Liquid-transport pipe-line is provided between road, the inner wall of the liquid-transport pipe-line is provided with hydrophobic coating.
The preferred embodiment of generating device as digital pcr microlayer model further includes sedimentation cavity, the sedimentation cavity setting
In the downstream of the ultrasonic atomizatio cavity, the microlayer model can settle in the sedimentation cavity.
The preferred embodiment of generating device as digital pcr microlayer model is provided with several settling legs in the sedimentation cavity,
The settling leg can assist the microlayer model to settle.
The surface of the preferred embodiment of generating device as digital pcr microlayer model, the settling leg is provided with hydrophobic coating.
The preferred embodiment of generating device as digital pcr microlayer model further includes exhaust outlet, one end of the exhaust outlet with
The sedimentation cavity is connected, and the other end is connected with outside atmosphere.
Beneficial effects of the present invention:
The generating device of digital pcr microlayer model of the invention, the device include micro-fluidic chip, power device and ultrasonic wave
Generator is provided with gas adding mouth, liquid adding mouth and ultrasonic atomizatio cavity, power device on micro-fluidic chip and is connected to gas
Body adding mouth and liquid adding mouth, gas sample and liquid sample are under the action of power device respectively from gas adding mouth and liquid
Body adding mouth enters in ultrasonic atomizatio cavity, and supersonic generator is arranged on the outside of ultrasonic atomizatio cavity, can be by liquid therein
The excitation of body sample is atomized into microlayer model.The device is based on ultrasonic atomizatio principle can form a large amount of microlayer models in a very short period of time, complete
The process of numerous droplet is distributed at digital pcr sample, generation speed is fast, and low energy consumption, and preparation facilities structure is simple, cost
It is low, solve the preparation methods such as silicon wafer micropore, high-precision spray head and PDMS chip equipment cost and technique it is more demanding and
The problems such as preparation time period length and cross contamination risk.
Detailed description of the invention
Fig. 1 is the top view of micro-fluidic chip provided by the invention；
Fig. 2 is the side view of micro-fluidic chip provided by the invention；
Fig. 3 is the top view of ultrasonic atomizatio cavity provided by the invention；
Fig. 4 is the side view of ultrasonic atomizatio cavity provided by the invention；
Fig. 5 is the top view of sedimentation cavity provided by the invention；
Fig. 6 is the side view of sedimentation cavity provided by the invention.
In figure:
1, gas adding mouth；2, liquid adding mouth；3, ultrasonic atomizatio cavity；4, cavity is settled；41, settling leg；5, gas-liquid
Mixing duct；6, liquid-transport pipe-line；7, flow resistance pipeline；8, exhaust outlet；9, microlayer model conveyance conduit；10, gas pipeline；11, it filters
Film；12, upper plate；13, lower plate；14, ultrasonic probe slot；15, ultrasonic probe.
Specific embodiment
Technical solution of the present invention is further illustrated with embodiment with reference to the accompanying drawing.It is understood that this place
The specific embodiment of description is used only for explaining the present invention rather than limiting the invention.It also should be noted that in order to
Convenient for description, in attached drawing, only the parts related to the present invention are shown and it is not all.
Present embodiment discloses a kind of generating device of digital pcr microlayer model, the generating device packets of the digital pcr microlayer model
Include micro-fluidic chip, power device and supersonic generator.As shown in Figure 1, be provided on micro-fluidic chip gas adding mouth 1,
Liquid adding mouth 2, ultrasonic atomizatio cavity 3 and sedimentation cavity 4, gas sample and liquid sample pass through gas by preset ratio respectively
Adding mouth 1 and liquid adding mouth 2 enter in ultrasonic atomizatio cavity 3.Power device is connected to gas adding mouth 1 and liquid adding mouth
2, power device can provide power for the flowing of gas sample and liquid sample.Supersonic generator is arranged in ultrasonic atomizatio chamber
Liquid sample in ultrasonic atomizatio cavity 3 can be excited and be atomized into microlayer model by 3 outside of body, supersonic generator.Settle cavity 4
It is connected with ultrasonic atomizatio cavity 3, microlayer model can settle in sedimentation cavity 4.
The device is based on ultrasonic atomizatio principle can form a large amount of microlayer models in a very short period of time, complete digital pcr sample point
It is scattered to the process of numerous droplet, generation speed is fast, and low energy consumption, and preparation facilities structure is simple, and it is at low cost, it is micro- to solve silicon wafer
The equipment cost and technique of the preparation methods such as hole, high-precision spray head and PDMS chip are more demanding and the preparation time period is long
And the problems such as cross contamination risk.
Power device includes the conventional use of constant pressure pump in micro-fluidic field, syringe pump and peristaltic pump etc., power device
Purpose is to provide power for gas sample and liquid sample, and the two is transported to the designated position on micro-fluidic chip.It needs
Illustrate no matter which kind of power device used, is both needed to using gas as power source and pushes liquid sample, i.e., by liquid sample
After being injected into liquid adding mouth 2, using gas pushes liquid sample to enter micro-fluidic chip.In order to guarantee power device and miniflow
The sealing on chip between adding mouth is controlled, can be realized by this fields conventional means such as female Luer, AB glue.Good sealing
It can guarantee to realize that gas sample and liquid sample are entered in micro-fluidic chip in the ratio of setting by controlling power device
In the structures such as each pipeline and ultrasonic atomizatio cavity 3.
In order to avoid generating pollution to PCR process, filter device is additionally provided between power device and adding mouth, on the one hand
Sample conveying can be had an impact to avoid the steam in power device, on the other hand also can avoid gas sample and carry the positive
Sample enters reaction system, to avoid generating pollution to PCR process.
The generating device of digital pcr microlayer model provided in this embodiment further includes gas-liquid mixed pipeline 5 and liquid-transport pipe-line 6,
The one end of one end and liquid adding mouth 2 far from power device of gas adding mouth 1 far from power device with gas-liquid mixed pipeline 5
Connection, the other end of gas-liquid mixed pipeline 5 are connected to ultrasonic atomizatio cavity 3.Above structure makes the setting of gas-liquid mixed pipeline 5 exist
Between gas adding mouth 1 and liquid adding mouth 2 and ultrasonic atomizatio cavity 3, in this way, gas sample and liquid sample are entering ultrasound
It can first be sufficiently mixed in gas-liquid mixed pipeline 5 before atomization cavity 3, be entered after ultrasonic atomizatio cavity 3 with improving
Nebulization efficiency and effect.Liquid-transport pipe-line 6 is provided between liquid adding mouth 2 and gas-liquid mixed pipeline 5, for conveying liquid-like
This.
As shown in Fig. 2, when in use, liquid sample is added to by liquid-transfering gun or other conventional liquid-transfering devices first
In cavity at liquid adding mouth 2, in order to avoid liquid sample rapidly enters liquid-transport pipe-line 6 and ultrasound under the action of capillary force
In atomization cavity 3, the inner wall of liquid-transport pipe-line 6 is provided with the drain valve that nanostructure is arranged in hydrophobic coating or liquid-transport pipe-line 6,
So that liquid sample be added to liquid adding mouth 2 after, without pressurization before, liquid sample can be stored in liquid sample-adding completely
In cavity at mouth 2.
Liquid sample in the present embodiment includes the necessary materials such as the necessary sample segment of PCR amplification, primer, enzyme, is
The stability of enhancing drop, also added certain surfactant, and surfactant can reduce droplet surface free energy,
Reduce the probability merged between drop, thus the drop rapid fusion for effectively avoiding atomization from generating.Surfactant is using number
The common PFPE-PEG-PFPE tripolymer in the field PCR is purchased from Ranbiotechnologie.Generally, liquid sample capacity
Range be 5 μ L-30 μ L, be capable of forming 5000-500000 microlayer models according to demand.
Pressure is applied to gas adding mouth 1 and liquid adding mouth 2 by power device, guarantees gas sample and liquid sample
It is collided in gas-liquid mixed pipeline 5 with the ratio of volume ratio 3:1-20:1 and appropriateness is mixed into gas-liquid mixture, above-mentioned volume ratio
It can be realized by the pressure or flow for adjusting power device.It can be set between gas adding mouth 1 and gas-liquid mixed pipeline 5
Flow resistance pipeline 7, it is therefore intended that increase flow resistance to the flowing of gas sample, so that it is guaranteed that the realization of aforementioned proportion, the flow resistance pipeline 7
If power device is sufficiently stable, or is capable of providing stable pressure proportional, it can not be set certainly for snake pipe
Set the flow resistance pipeline 7.
As shown in figs 2-4, it is provided at position corresponding with the middle part of ultrasonic atomizatio cavity 3 on micro-fluidic chip super
Sonic probe slot 14, ultrasonic probe slot 14 are used to accommodate the ultrasonic probe 15 of supersonic generator.The middle part of ultrasonic atomizatio cavity 3 and
The depth H 2 at the position of the ultrasonic probe 15 contact is greater than the arrival end of ultrasonic atomizatio cavity 3 and the depth H 1 of outlet end, therefore
Liquid in above-mentioned gas-liquid mixture can be attached to cavity bottom since gravity settles, i.e., attached close to ultrasonic probe slot 14
Closely.After starting supersonic generator, ultrasonic wave occurs for ultrasonic probe 15, the liquid in ultrasonic atomizatio cavity 3 can be excited mist
Change, quickly generates microlayer model.It should be noted that the efficiency in order to guarantee ultrasonic atomizatio, it is required that the face of ultrasonic probe 15
The most areas of product covering ultrasonic probe slot 14, to avoid there are big quantity of fluid due to can not receive ultrasonic energy not by
It is atomized into microlayer model.In the present embodiment, the value range of H2 is 1mm-3mm, and the value range of H1 is 0.05mm-1mm, can basis
Design requirement, working ability and material property are designed.
The arrival end of ultrasonic atomizatio cavity 3 and outlet end pass through inclined-plane and connect with middle part, to avoid the unexpected change of depth
Change forms dead volume.
The diameter for the microlayer model that ultrasonic atomizatio is formed is adjustable, and diameter can be estimated by following empirical equation: D=0.34
(8 π T/ ρ f2) 1/3, D is liquid-drop diameter, and T is surface tension coefficient, and ρ is fluid density, and f is sound wave rate.
According to the property of above-mentioned formula and common liquid, the supersonic generator selection in the present embodiment is common on the market
Hyperfrequency supersonic generator, the frequency range of supersonic generator are 104Hz-106Hz can satisfy 10 μm -60 of liquid-drop diameter
μm requirement.The power bracket of supersonic generator is 2W-20W in the present embodiment, and specific value needs reference fluids property, chip
The thickness of material and ultrasonic probe slot 14.
The material of micro-fluidic chip includes silicon wafer, glass, PDMS, PMMA, PC and PS etc., and processing method can be according to material
Performance selects the methods of photoetching, wet etching, Soft lithograph, numerical control processing, laser processing, injection molding or hot pressing.Wherein it should be noted that
, the intrinsic physical characteristic of material will affect the transmitting of 15 energy of ultrasonic probe, therefore, in the material of selection micro-fluidic chip
When, on the one hand need to guarantee that the material is not crushed in ultrasonication frequency, it on the other hand need to be according to the property of material itself
It is adjustable the frequency of ultrasonic atomizatio, guarantees the generation diameter and rate of microlayer model.
The downstream connection of ultrasonic atomizatio cavity 3 has sedimentation cavity 4, is connected between the two by microlayer model conveyance conduit 9, mist
After change.The microlayer model that formation is atomized in ultrasonic atomizatio cavity 3 is suspended in the top of cavity, and power device continues gas at this time
Body injects in chip, and when gas passes through ultrasonic atomizatio cavity 3, the microlayer model for driving atomization to be formed passes through microlayer model conveyance conduit 9,
It enters in sedimentation cavity 4.The downstream of sedimentation cavity 4 is communicated with exhaust outlet 8 by gas pipeline 10, and exhaust outlet 8 and outside are big
Gas phase connection.
Referring to shown in Fig. 1, Fig. 2, Fig. 5 and Fig. 6, the structure type of sedimentation cavity 4 is similar to ultrasonic atomizatio cavity 3, chamber
After body middle part is wider relatively deep, therefore the mixture of the microlayer model and gas after atomization enters sedimentation cavity 4, the movement rate of gas
Become smaller, gravity plays main function, and microlayer model is deposited in sedimentation 4 bottom of cavity, and gas then under the action of power device, passes through
Gas pipeline 10 is discharged from exhaust outlet 8.In order to avoid residual liquid pollutes the environment, it was provided at exhaust outlet 8
Filter membrane 11, can be by all filtering interceptions of remaining microlayer model.
Further, settling in cavity 4 can be set several settling legs 41, the top and bottom of settling leg 41 with
The inner wall of cavity connects.Settling leg 41 can effectively stop the flowing of microlayer model, promote microlayer model further to settle, avoid micro- liquid
Drop enters gas pipeline 10.Settling leg 41 in the present embodiment is set as the form of rectangular array, and the resistance to microlayer model can be improved
Gear rate.
Preferably, hydrophobic coating can be arranged in the bottom surface of sedimentation cavity 4 according to demand, to reduce the fusion of microlayer model
Probability can also prestore a certain amount of oily phase, be attached to oily phase surface after settling microlayer model, reduce the free energy of microlayer model, oil
It mutually can be fluorocarbon oil commonly used in the art.
The surface of microlayer model conveyance conduit 9, settling leg 41 and gas pipeline 10 is provided with hydrophobic coating, to reduce micro- liquid
The absorption of drop avoids microlayer model from largely adsorbing in said structure.
Settle the depth H 3 and above-mentioned flow resistance pipeline 7, gas-liquid mixed pipeline 5, liquid-transport pipe-line 6, microlayer model of 4 inlet of cavity
Conveyance conduit 9 and gas pipeline 10 it is substantially uniform-diameter, i.e., the value range of H3 is 0.05mm-1.0mm, and specific value needs root
It is selected according to design requirement, working ability and material property；The depth H 4 for settling 4 middle part of cavity is larger, and value range is
1mm-3mm；It is similar to 3 structure of ultrasonic atomizatio cavity, it settles and is provided with ramp structure at the entrance and exit of cavity 4, avoid
The suddenly change of depth forms dead volume.
In sedimentation cavity 4, the sample of digital pcr is wrapped in the process knot in microlayer model for most microlayer model final settlements
Beam can be required according to subsequent operation, temperature control plate and detection device be arranged above or below 4 outside of sedimentation cavity, thus complete
At sample preparation-amplification of entire digital pcr and detection process.
In the present embodiment, micro-fluidic chip includes upper plate 12 and lower plate 13, above-mentioned gas adding mouth 1, liquid adding mouth
2, exhaust outlet 8 and filter membrane 11 are processed on upper plate 12, ultrasonic atomizatio cavity 3, sedimentation cavity 4, gas-liquid mixed pipeline 5, defeated
Liquid pipe road 6, flow resistance pipeline 7, drop conveyance conduit 9 and gas pipeline 10 can be according to design and manufacturing process difference, selection processing
In upper plate 12 or lower plate 13 or simultaneous processing on upper plate 12 and lower plate 13.
The present embodiment is wider using the microlayer model particle diameter distribution that ultrasonic atomizatio mode is prepared, which meets typical
Poisson distribution, influence factor mainly include liquid property (viscosity, interfacial tension), 15 property of ultrasonic probe (power, frequency,
Size), (PMMA and PDMS damping are different, and same supersonic frequency mists with material for chip size (depth of ultrasonic probe slot 14)
It is different to change liquid-drop diameter) etc., the median of the diameter Poisson distribution of microlayer model can be set as examining by adjusting above-mentioned parameter
Survey the optimal value for the microlayer model diameter that device can detect.In addition, having DNA fragmentation to be detected, DNA piece in liquid sample
Poisson distribution is also obeyed in distribution of the section in microlayer model.Need by adjust liquid sample property (emphasis includes that DNA fragmentation is dense
Degree), 15 property of ultrasonic probe, chip size and material, to realize in most microlayer models within the scope of detection line, at most only
An a DNA fragmentation to be detected, meets the requirement of digital pcr.
To detect mutation type p.L858R lung cancer, for target gene is EGFR, commercially available EGFR (L858R) examination is purchased
Agent box, after sample solution is carried out necessary processing and nucleic acid extraction, according to kit specification, by sample nucleic acid solution and reagent
The reagents such as enzyme, primer, probe in box mix in proportion, and liquid sample is made.Pay close attention to the dense of DNA fragmentation in sample solution
Degree guarantees the microlayer model for meeting detection range diameter, and the amount of droplets of only one of them DNA fragmentation to be measured meets system need
It asks.
Obviously, the above embodiment of the present invention is just for the sake of clearly illustrating examples made by the present invention, and being not is pair
The restriction of embodiments of the present invention.For those of ordinary skill in the art, may be used also on the basis of the above description
To make other variations or changes in different ways.There is no necessity and possibility to exhaust all the enbodiments.It is all this
Made any modifications, equivalent replacements, and improvements etc., should be included in the claims in the present invention within the spirit and principle of invention
Protection scope within.
Claims (10)
1. a kind of generating device of digital pcr microlayer model characterized by comprising
Micro-fluidic chip is provided with gas adding mouth (1), liquid adding mouth (2) and ultrasonic atomizatio cavity (3), gas sample
This is passed through the gas adding mouth (1) with liquid sample respectively by preset ratio and the liquid adding mouth (2) enters described surpass
In sound atomization cavity (3)；
Power device, is connected to the gas adding mouth (1) and liquid adding mouth (2), and the power device can be described
The flowing of gas sample and the liquid sample provides power；
Supersonic generator, on the outside of the ultrasonic atomizatio cavity (3), the supersonic generator described can will surpass for setting
Liquid sample excitation in sound atomization cavity (3) is atomized into microlayer model.
2. the generating device of digital pcr microlayer model according to claim 1, which is characterized in that the ultrasonic atomizatio cavity
(3) depth at middle part is greater than the arrival end of the ultrasonic atomizatio cavity (3) and the depth of outlet end.
3. the generating device of digital pcr microlayer model according to claim 2, which is characterized in that the arrival end and described
Outlet end passes through inclined-plane and connect with the middle part, and the inclined-plane can be avoided the formation of dead volume.
4. the generating device of digital pcr microlayer model according to claim 2, which is characterized in that the micro-fluidic chip with
It is provided with ultrasonic probe slot (14) at the corresponding position in middle part of the ultrasonic atomizatio cavity (3), the ultrasonic probe slot
(14) ultrasonic probe (15) of the supersonic generator can be accommodated.
5. the generating device of digital pcr microlayer model according to claim 1, which is characterized in that further include gas-liquid mixed pipe
Road (5), the gas adding mouth (1) one end and the liquid adding mouth (2) far from the power device are far from the power
One end of device is connected to the gas-liquid mixed pipeline (5), the other end of the gas-liquid mixed pipeline (5) and the ultrasonic mist
Change cavity (3) connection.
6. the generating device of digital pcr microlayer model according to claim 5, which is characterized in that the liquid adding mouth (2)
It is provided with liquid-transport pipe-line (6) between the gas-liquid mixed pipeline (5), the inner wall of the liquid-transport pipe-line (6) is provided with hydrophobic painting
Layer.
7. the generating device of digital pcr microlayer model according to claim 1, which is characterized in that further include sedimentation cavity
(4), sedimentation cavity (4) is arranged in the downstream of the ultrasonic atomizatio cavity (3), and the microlayer model can be in the sedimentation chamber
It is settled in body (4).
8. the generating device of digital pcr microlayer model according to claim 7, which is characterized in that in the sedimentation cavity (4)
It is provided with several settling legs (41), the settling leg (41) can assist the microlayer model to settle.
9. the generating device of digital pcr microlayer model according to claim 8, which is characterized in that the settling leg (41)
Surface is provided with hydrophobic coating.
10. the generating device of digital pcr microlayer model according to claim 7, which is characterized in that it further include exhaust outlet (8),
One end of the exhaust outlet (8) is connected with the sedimentation cavity (4), and the other end is connected with outside atmosphere.
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